Anne E Cress

Interests

Teaching

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Research

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Courses

Scholarly Contributions

Books

• Cress, A. E. (2006). Cell Adhesion and Cytoskeletal Molecules in Metastasis.

Chapters

• Algotar, A. M., Cress, A. E., & Algotar, A. E. (2019). Prostate Cancer Prevention. In Cancer Prevention. Springer International Publishing. doi:10.1007/978-3-030-15935-1_17
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  Prostate cancer is a major cause of morbidity and mortality for American men. Hence, it is essential to understand the factors that could be helpful in preventing it. Majority of incident cancer follows an indolent trajectory, although significant percent follows an aggressive trajectory leading to considerable morbidity and mortality. Due to this, the emphasis of the current chapter is on secondary rather than primary prevention. Currently available diagnostic modalities are unable to reliably distinguish between aggressive and indolent disease. Hence, efforts need to be made to understand the factor associated with secondary and tertiary prevention of prostate cancer. This chapter systematically reviews literature starting with the changing guidelines, the modifiable risk factors for prostate cancer prevention, factors affecting prostate cancer survivorship, novel avenues for prostate cancer prevention, and finishing with themes to be addressed in the future that could be instrumental in preventing prostate cancer.
• Algotar, A. M., Harryman, W. L., Cress, A. E., & Stratton, M. S. (2014). Prevention of Prostate Cancer. In Cancer Prevention. Springer Berlin Heidelberg. doi:10.1007/978-3-642-38983-2_16
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  According to the American Cancer Society, prostate cancer is the second most common cause of cancer-related mortality for US men. A continuing research challenge is to improve diagnosis (especially the ability to detect uncommon aggressive cancers) and improve chemoprevention (through nutrition and lifestyle factors). The chapter highlights the growing importance of secondary prevention since prostate cancer is an indolent, slow growing tumor; preventing early tumor progression or eradicating early bone metastases may cure the disease. Considerations of the known etiology and biology of prostate cancer are presented as well as the strengths and limitations of current diagnostic and prognostic factors and specific interventions that have been tested. Lastly, promising avenues for secondary prevention strategies are presented based upon current knowledge.
• Davis, T. L., Goldman, A., Cress, A. E., & Davis, T. P. (2006). SUPPRESSION AND ALTERATION OF ADHESION STRUCTURES IN HUMAN EPITHELIAL CANCER PROGRESSION. In Cell Adhesion and Cytoskeletal Molecules in Metastasis. Springer, Dordrecht. doi:10.1007/978-1-4020-5129-6_2
• Sroka, T., Cress, A. E., & Lam, K. S. (2006). EPITHELIAL CELL SURFACE TARGETING USING SYNTHETIC D-AMINO ACID PEPTIDES. In Cell Adhesion and Cytoskeletal Molecules in Metastasis. Springer, Dordrecht. doi:10.1007/978-1-4020-5129-6_10

Journals/Publications

• Lee, B. R., Wong, A. C., Pollock, G., Cress, A. E., Batai, K., Warfel, N. A., Ignatenko, N., & Maar, K. D. (2023). Digital Image analysis using Video microscopy of human-derived prostate cancer vs normaprostate organoids to assess migratory behavior on extracellular matrix proteins.. Frontiers in Oncology.
• Warfel, N. A., Rogers, G. C., Mouneimne, G., Miranti, C., Cress, A. E., Kraft, A., Daulat, S. R., Casillas, A. L., Chauhan, S. S., Toth, R. K., Langlais, P. R., Bethard, J. R., Ball, L. E., Liou, H., Clements, A. N., & Jensen, C. (2023). PIM kinases regulate actin dynamics and tumor cell invasion in hypoxia. Journal of Cell Biology.
• Cress, A. E., Moreno-Vinasco, L., Casanova, N. G., Garcia, A. N., Kempf, C. L., Bermudez, T., Valera, D. G., Song, J. H., Sun, X., Cai, H., Gregory, T., Oita, R. C., Hernon, V. R., Camp, S. M., Rogers, C., Kyubwa, E. M., Menon, N., Axtelle, J., Rappaport, J., , Bime, C., et al. (2022). eNAMPT Is a Novel Damage-associated Molecular Pattern Protein That Contributes to the Severity of Radiation-induced Lung Fibrosis. American Journal of Respiratory Cell and Molecular Biology, 66(5), 497-509. doi:10.1165/rcmb.2021-0357oc
• Mascarenhas, J. B., Cress, A. E., Garcia, J. G., Dudek, S. M., Camp, S. M., Jacobson, J. R., Gaber, A. A., & Song, J. H. (2022). An Actin-, Cortactin- and Ena-VASPLinked Complex Contributes to Endothelial Cell Focal Adhesion and Vascular Barrier Regulation. Cellular Physiology and Biochemistry. doi:10.33594/000000553
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  Increase in vascular permeability is a cardinal feature of all inflammatory diseases and represents an imbalance in vascular contractile forces and barrier-restorative forces, both of which are highly dependent on actin cytoskeletal dynamics. In addition to the involvement of key vascular barrier-regulatory, actin-binding proteins, such as nmMLCK and cortactin, we recently demonstrated a role for a member of the Ena-VASP family known as Ena-VASP-like (EVL) in promoting vascular focal adhesion (FA) remodeling and endothelial cell (EC) barrier restoration/preservation.To further understand the role of EVL in EC barrier-regulatory processes, we examined EVL-cytoskeletal protein interactions in FA dynamics in vitro utilizing lung EC and in vivo murine models of acute inflammatory lung injury. Deletion mapping studies and immunoprecipitation assays were performed to detail the interaction between EVL and cortactin, and further evaluated by assessment of changes in vascular EC permeability following disruption of EVL-cortactin interaction.Initial studies focusing on the actin-binding proteins, nmMLCK and cortactin, utilized deletion mapping of the cortactin gene (CTTN) to identify cortactin domains critical for EVL-cortactin interaction and verified the role of actin in promoting EVL-cortactin interaction. A role for profilins, actin-binding proteins that regulate actin polymerization, was established in facilitating EVL-FA binding.In summary, these studies further substantiate EVL participation in regulation of vascular barrier integrity and in the highly choreographed cytoskeletal interactions between key FA and cytoskeletal partners.
• Guerra, S., David, J. M., Younis, U. S., Whalen, M. B., Romanoski, C. E., Polverino, F., Pilon, A. L., Menghani, S. V., Martinez, F. D., Ledford, J. G., Kraft, M., Johnson, M. D., Guerra, S., David, J. M., Cress, A. E., & Addison, K. J. (2021). CC16 Binding to α4β1 Integrin Protects against Mycoplasma pneumoniae Infection.. American journal of respiratory and critical care medicine, 203(11), 1410-1418. doi:10.1164/rccm.202006-2576oc
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  Rationale CC16 (club cell secretory protein) is a pneumoprotein produced predominantly by pulmonary club cells. Circulating CC16 is associated with protection from the inception and progression of the two most common obstructive lung diseases (asthma and chronic obstructive pulmonary disease). Objectives Although exact mechanisms remain elusive, studies consistently suggest a causal role of CC16 in mediating antiinflammatory and antioxidant functions in the lung. We sought to determine any novel receptor systems that could participate in CC16's role in obstructive lung diseases. Methods Protein alignment of CC16 across species led to the discovery of a highly conserved sequence of amino acids, leucine-valine-aspartic acid (LVD), a known integrin-binding motif. Recombinant CC16 was generated with and without the putative integrin-binding site. A Mycoplasma pneumoniae mouse model and a fluorescent cellular adhesion assay were used to determine the impact of the LVD site regarding CC16 function during live infection and on cellular adhesion during inflammatory conditions. Measurements and Main Results CC16 bound to integrin α4β1), also known as the adhesion molecule VLA-4 (very late antigen 4), dependent on the presence of the LVD integrin-binding motif. During infection, recombinant CC16 rescued lung function parameters both when administered to the lung and intravenously but only when the LVD integrin-binding site was intact; likewise, neutrophil recruitment during infection and leukocyte adhesion were both impacted by the loss of the LVD site. Conclusions We discovered a novel receptor for CC16, VLA-4, which has important mechanistic implications for the role of CC16 in circulation as well as in the lung compartment.
• Harryman, W. L., Marr, K. D., Hernandez-Cortes, D., Nagle, R. B., Garcia, J. G., & Cress, A. E. (2021). Cohesive cancer invasion of the biophysical barrier of smooth muscle. Cancer metastasis reviews.
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  Smooth muscle is found around organs in the digestive, respiratory, and reproductive tracts. Cancers arising in the bladder, prostate, stomach, colon, and other sites progress from low-risk disease to high-risk, lethal metastatic disease characterized by tumor invasion into, within, and through the biophysical barrier of smooth muscle. We consider here the unique biophysical properties of smooth muscle and how cohesive clusters of tumor use mechanosensing cell-cell and cell-ECM (extracellular matrix) adhesion receptors to move through a structured muscle and withstand the biophysical forces to reach distant sites. Understanding integrated mechanosensing features within tumor cluster and smooth muscle and potential triggers within adjacent adipose tissue, such as the unique damage-associated molecular pattern protein (DAMP), eNAMPT (extracellular nicotinamide phosphoribosyltransferase), or visfatin, offers an opportunity to prevent the first steps of invasion and metastasis through the structured muscle.
• Okumura, K., Kraft, A., Buchan, J. R., Nelson, A., Cress, A. E., Peti, W., Gard, J., Harter, M., Li, Y., Fernandes, N., Mouneimne, G., Song, J., Cardo Vila, M., Singh, N., Padi, S., & Bearss, J. (2021). EDC3 Phosphorylation Regulates Growth and Invasion through controlling P-body Formation and Dynamics. EMBO Report.
• Okumura, K., Okumura, K., Okumura, K., Kraft, A., Kraft, A., Kraft, A., Buchan, J. R., Buchan, J. R., Buchan, J. R., Nelson, A., Nelson, A., Nelson, A., Cress, A. E., Cress, A. E., Cress, A. E., Peti, W., Peti, W., Peti, W., Gard, J., , Gard, J., et al. (2021). EDC3 Phosphorylation Regulates Growth and Invasion through controlling P-body Formation and Dynamics. EMBO Report.
• Sun, B., Sammani, S., Martin, D., Desai, A. A., Valera, D. G., Sun, X., Sun, B., Song, J. H., Sammani, S., Quijada, H., Oita, R. C., Natarajan, V., Moreno-vinasco, L., Mascarenhas, J. B., Martin, D. R., Liu, Z., Kempf, C. L., Jacobson, J. R., Hernon, V. R., , Garcia, J. G., et al. (2021). Endothelial eNAMPT amplifies pre-clinical acute lung injury: efficacy of an eNAMPT-neutralising monoclonal antibody.. The European respiratory journal, 57(5). doi:10.1183/13993003.02536-2020
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  The severe acute respiratory syndrome coronavirus 2/coronavirus disease 2019 pandemic has highlighted the serious unmet need for effective therapies that reduce acute respiratory distress syndrome (ARDS) mortality. We explored whether extracellular nicotinamide phosphoribosyltransferase (eNAMPT), a ligand for Toll-like receptor (TLR)4 and a master regulator of innate immunity and inflammation, is a potential ARDS therapeutic target..Wild-type C57BL/6J or endothelial cell (EC)-cNAMPT -/- knockout mice (targeted EC NAMPT deletion) were exposed to either a lipopolysaccharide (LPS)-induced ("one-hit") or a combined LPS/ventilator ("two-hit")-induced acute inflammatory lung injury model. A NAMPT-specific monoclonal antibody (mAb) imaging probe (99mTc-ProNamptor) was used to detect NAMPT expression in lung tissues. Either an eNAMPT-neutralising goat polyclonal antibody (pAb) or a humanised monoclonal antibody (ALT-100 mAb) were used in vitro and in vivo..Immunohistochemical, biochemical and imaging studies validated time-dependent increases in NAMPT lung tissue expression in both pre-clinical ARDS models. Intravenous delivery of either eNAMPT-neutralising pAb or mAb significantly attenuated inflammatory lung injury (haematoxylin and eosin staining, bronchoalveolar lavage (BAL) protein, BAL polymorphonuclear cells, plasma interleukin-6) in both pre-clinical models. In vitro human lung EC studies demonstrated eNAMPT-neutralising antibodies (pAb, mAb) to strongly abrogate eNAMPT-induced TLR4 pathway activation and EC barrier disruption. In vivo studies in wild-type and EC-cNAMPT -/- mice confirmed a highly significant contribution of EC-derived NAMPT to the severity of inflammatory lung injury in both pre-clinical ARDS models..These findings highlight both the role of EC-derived eNAMPT and the potential for biologic targeting of the eNAMPT/TLR4 inflammatory pathway. In combination with predictive eNAMPT biomarker and NAMPT genotyping assays, this offers the opportunity to identify high-risk ARDS subjects for delivery of personalised medicine.
• Warfel, N. A., Cress, A. E., Miranti, C. K., Langlais, P. R., Jensen, C. C., Clements, A. N., Sainz, A. G., Toth, R. K., Chauhan, S. S., & Casillas, A. L. (2021). Direct phosphorylation and stabilization of HIF-1a by PIM1 kinase drives angiogenesis in solid tumors. Oncogene.
• Warfel, N. A., Cress, A. E., Miranti, C. K., Langlais, P. R., Jensen, C. C., Clements, A. N., Toth, R. K., Chauhan, S. S., & Casillas, A. L. (2021). Direct phosphorylation and stabilization of HIF-1a by PIM1 kinase drives angiogenesis in solid tumors. Cell Reports.
• Johnson, M. D., Younis, U. S., Menghani, S. V., Addison, K. J., Whalen, M., Pilon, A. L., Cress, A. E., Polverino, F., Romanoski, C., Kraft, M., Martinez, F. D., Guerra, S., & Ledford, J. G. (2020). CC16 Binding to α4β1 Integrin (VLA-4) Protects Against Infection. American journal of respiratory and critical care medicine.
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  Club cell secretory protein (CC16) is a pneumoprotein produced predominantly by pulmonary club cells. Circulating CC16 is associated with protection from the inception and progression of the two most common obstructive lung diseases: asthma and COPD. While exact mechanisms remain elusive, studies consistently suggest a causal role of CC16 in mediating anti-inflammatory and antioxidant functions in the lung. We sought to determine any novel receptor systems that could participate in CC16's role in obstructive lung diseases. Protein alignment of CC16 across species led to the discovery of a highly conserved sequence of amino acids, Leucine-Valine-Aspartic Acid (LVD), a known integrin binding motif. Recombinant CC16 was generated with and without the putative integrin binding site. A Mycoplasma pneumoniae mouse model and a flourescent cellular adhesion assay were used to determine the impact of the LVD site in regards to CC16 function during live infection and on cellular adhesion during inflammatory conditions. CC16 bound to integrin alpha 4 and beta 1 (α4β1), also known as the adhesion molecule very late antigen-4 (VLA-4), dependent on the presence of the LVD integrin binding motif. During infection, rCC16 rescued lung function parameters both when administered to the lung and intravenously, but only when the LVD integrin binding site is intact; likewise, neutrophil recruitment during infection and leukocyte adhesion were both impacted by the loss of the LVD site. We discovered a novel receptor for CC16, VLA-4, which has important mechanistic implications for the role of CC16 in circulation as well as in the lung compartment.
• Kelly, G. T., Faraj, R., Dai, Z., Cress, A. E., & Wang, T. (2020). A mutation found in esophageal cancer alters integrin β4 mRNA splicing. Biochemical and biophysical research communications, 529(3), 726-732.
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  Integrin β4 (CD104, mRNA: ITGβ4) contributes to anchoring cells to the extracellular matrix and is regulated in many cancer types where it contributes to tumor progression. One splice variant, integrin β4E, is poorly characterized. We extracted several mutations from tumor samples within ITGB4 near the splice site that controls ITGβ4E production, and computational analysis predicted six of these would alter splicing to alter ITGβ4E abundance. One of these mutations, from an esophageal squamous cell carcinoma sample, was predicted to increase splicing toward ITGβ4E. We verified this effect using a minigene, and observed that integrin β4E slows esophageal squamous cell migration while other variants enhance migration, demonstrating that integrin β4E regulation through mutations may contribute to esophageal squamous cell tumorigenesis.
• Lee, B. R., Wong, A. C., Warfel, N. A., Pollock, G. R., Marr*, K., Marr, K. D., Lee, B. R., Ignatenko, N., Cress, A. E., & Batai, K. (2020). MP09-07 USE OF 3D VIDEO MICROSCOPY OF HUMAN-DERIVED PROSTATE ORGANOIDS TO ASSESS FEATURES OF EARLY INVASIVE PROSTATE CANCER. The Journal of Urology, 203, e118. doi:10.1097/ju.0000000000000829.07
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  INTRODUCTION AND OBJECTIVE:Patient-derived organoids provide a method for creating human prostate cancer cell lines in culture in three dimensions. We established living organoids using normal and ...
• Miranti, C. K., Tran, J. D., Schulz, V. V., Nollet, E. A., Miranti, C. K., Ganguly, S. S., Cress, A. E., Corey, E., & Cardo-vila, M. (2020). Androgen receptor-induced integrin α6β1 and Bnip3 promote survival and resistance to PI3K inhibitors in castration-resistant prostate cancer.. Oncogene, 39(31), 5390-5404. doi:10.1038/s41388-020-1370-9
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  The androgen receptor (AR) is the major driver of prostate cancer growth and survival. However, almost all patients relapse with castration-resistant disease (CRPC) when treated with anti-androgen therapy. In CRPC, AR is often aberrantly activated independent of androgen. Targeting survival pathways downstream of AR could be a viable strategy to overcome CRPC. Surprisingly, little is known about how AR drives prostate cancer survival. Furthermore, CRPC tumors in which Pten is lost are also resistant to eradication by PI3K inhibitors. We sought to identify the mechanism by which AR drives tumor survival in CRPC to identify ways to overcome resistance to PI3K inhibition. We found that integrins α6β1 and Bnip3 are selectively elevated in CRPC downstream of AR. While integrin α6 promotes survival and is a direct transcriptional target of AR, the ability of AR to induce Bnip3 is dependent on adhesion to laminin and integrin α6β1-dependent nuclear translocation of HIF1α. Integrins α6β1 and Bnip3 were found to promote survival of CRPC cells selectively on laminin through the induction of autophagy and mitophagy. Furthermore, blocking Bnip3 or integrin α6β1 restored sensitivity to PI3K inhibitors in Pten-negative CRPC. We identified an AR driven pathway that cooperates with laminin and hypoxia to drive resistance to PI3K inhibitors. These findings can help explain in part why PI3K inhibitors have failed in clinical trials to overcome AR-dependent CRPC.
• Quijada, H., Bermudez, T., Kempf, C. L., Valera, D. G., Garcia, A. N., Camp, S. M., Song, J. H., Franco, E., Burt, J. K., Sun, B., Mascarenhas, J. B., Burns, K., Gaber, A., Oita, R. C., Reyes Hernon, V., Barber, C., Moreno-Vinasco, L., Sun, X., Cress, A. E., , Martin, D., et al. (2020). Endothelial eNAMPT Amplifies Preclinical Acute Lung Injury: Efficacy of an eNAMPT-Neutralising mAb. The European respiratory journal.
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  The SARS-CoV-2/COVID-19 pandemic has highlighted the serious unmet need for effective therapies that reduce ARDS mortality. We explored whether extracellular nicotinamide phosphoribosyltransferase (eNAMPT), a ligand for Toll-like receptor 4 and a master regulator of innate immunity and inflammation, is a potential ARDS therapeutic target.
• Sun, B. L., Sun, X., Casanova, N., Garcia, A. N., Oita, R., Algotar, A. M., Camp, S. M., Hernon, V. R., Gregory, T., Cress, A. E., & Garcia, J. G. (2020). Role of secreted extracellular nicotinamide phosphoribosyltransferase (eNAMPT) in prostate cancer progression: Novel biomarker and therapeutic target. EBioMedicine, 61, 103059.
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  There remains a serious need to prevent the progression of invasive prostate cancer (PCa). We previously showed that secreted extracellular nicotinamide phosphoribosyltransferase (eNAMPT) is a multifunctional innate immunity regulator via TLR4 ligation which has been implicated in PCa progression. Here we investigate the role of eNAMPT as a diagnostic biomarker and therapeutic target in the progression of PCa.
• Wang, M., Nagle, R. B., Knudsen, B. S., Cress, A. E., & Rogers, G. C. (2020). Centrosome loss results in an unstable genome and malignant prostate tumors. Oncogene, 39(2), 399-413.
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  Localized, nonindolent prostate cancer (PCa) is characterized by large-scale genomic rearrangements, aneuploidy, chromothripsis, and other forms of chromosomal instability (CIN), yet how this occurs remains unclear. A well-established mechanism of CIN is the overproduction of centrosomes, which promotes tumorigenesis in various mouse models. Therefore, we developed a single-cell assay for quantifying centrosomes in human prostate tissue. Surprisingly, centrosome loss-which has not been described in human cancer-was associated with PCa progression. By chemically or genetically inducing centrosome loss in nontumorigenic prostate epithelial cells, mitotic errors ensued, producing aneuploid, and multinucleated cells. Strikingly, transient or chronic centrosome loss transformed prostate epithelial cells, which produced highly proliferative and poorly differentiated malignant tumors in mice. Our findings suggest that centrosome loss could create a cellular crisis with oncogenic potential in prostate epithelial cells.
• Cress, A. E. (2019). A Method to Reuse Archived H&E Stained Histology Slides for a Multiplex Protein Biomarker Analysis. Methods and Protocols.
• Cress, A. E. (2019). A method of quantifying centrosomes at the single-cell level in human normal and cancer tissue.. Molecular biology of the cell.
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  Centrosome abnormalities are emerging hallmarks of cancer. The overproduction of centrosomes (known as centrosome amplification) has been reported in a variety of cancers and is currently being explored as a promising target for therapy. However, to understand different types of centrosome abnormalities and their impact on centrosome function during tumor progression, as well as to identify tumor subtypes that would respond to the targeting of a centrosome abnormality, a reliable method for accurately quantifying centrosomes in human tissue samples is needed. Here, we established a method of quantifying centrosomes at a single-cell level in different types of human tissue samples. We tested multiple anti-centriole and pericentriolar-material antibodies to identify bona fide centrosomes and multiplexed these with cell border markers to identify individual cells within the tissue. High-resolution microscopy was used to generate multiple Z-section images, allowing us to acquire whole cell volumes in which to scan for centrosomes. The normal cells within the tissue serve as internal positive controls. Our method provides a simple, accurate way to distinguish alterations in centrosome numbers at the level of single cells.
• Cress, A. E. (2019). Centrosome loss results in an unstable genome and malignant prostate tumors.. Oncogene.
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  Localized, nonindolent prostate cancer (PCa) is characterized by large-scale genomic rearrangements, aneuploidy, chromothripsis, and other forms of chromosomal instability (CIN), yet how this occurs remains unclear. A well-established mechanism of CIN is the overproduction of centrosomes, which promotes tumorigenesis in various mouse models. Therefore, we developed a single-cell assay for quantifying centrosomes in human prostate tissue. Surprisingly, centrosome loss-which has not been described in human cancer-was associated with PCa progression. By chemically or genetically inducing centrosome loss in nontumorigenic prostate epithelial cells, mitotic errors ensued, producing aneuploid, and multinucleated cells. Strikingly, transient or chronic centrosome loss transformed prostate epithelial cells, which produced highly proliferative and poorly differentiated malignant tumors in mice. Our findings suggest that centrosome loss could create a cellular crisis with oncogenic potential in prostate epithelial cells.
• Cress, A. E. (2019). Gene Editing of α6 Integrin Inhibits Muscle Invasive Networks and Increases Cell-Cell Biophysical Properties in Prostate Cancer.. Cancer research.
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  Human prostate cancer confined to the gland is indolent (low-risk), but tumors outside the capsule are aggressive (high-risk). Extracapsular extension requires invasion within and through a smooth muscle-structured environment. Because integrins respond to biomechanical cues, we used a gene editing approach to determine if a specific region of laminin-binding α6β1 integrin was required for smooth muscle invasion both in vitro and in vivo. Human tissue specimens showed prostate cancer invasion through smooth muscle and tumor coexpression of α6 integrin and E-cadherin in a cell-cell location and α6 integrin in a cell-extracellular matrix (ECM) distribution. Prostate cancer cells expressing α6 integrin (DU145 α6WT) produced a 3D invasive network on laminin-containing Matrigel and invaded into smooth muscle both in vitro and in vivo. In contrast, cells without α6 integrin (DU145 α6KO) and cells expressing an integrin mutant (DU145 α6AA) did not produce invasive networks, could not invade muscle both in vitro and in vivo, and surprisingly formed 3D cohesive clusters. Using electric cell-substrate impedance testing, cohesive clusters had up to a 30-fold increase in normalized resistance at 400 Hz (cell-cell impedance) as compared with the DU145 α6WT cells. In contrast, measurements at 40,000 Hz (cell-ECM coverage) showed that DU145 α6AA cells were two-fold decreased in normalized resistance and were defective in restoring resistance after a 1 μmol/L S1P challenge as compared with the DU145 α6WT cells. The results suggest that gene editing of a specific α6 integrin extracellular region, not required for normal tissue function, can generate a new biophysical cancer phenotype unable to invade the muscle, presenting a new therapeutic strategy for metastasis prevention in prostate cancer. SIGNIFICANCE: This study shows an innovative strategy to block prostate cancer metastasis and invasion in the muscle through gene editing of a specific α6 integrin extracellular region.
• Cress, A. E. (2019). Integrin α6β4E variant is associated with actin and CD9 structures and modifies the biophysical properties of cell-cell and cell-extracellular matrix interactions.. Molecular biology of the cell.
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  Integrin α6β4 is an essential, dynamic adhesion receptor for laminin 332 found on epithelial cells, required for formation of strong cell-extracellular matrix (ECM) adhesion and induced migration, and coordinated by regions of the β4C cytoplasmic domain. β4E, a unique splice variant of β4 expressed in normal tissue, contains a cytoplasmic domain of 231 amino acids with a unique sequence of 114 amino acids instead of β4C's canonical 1089 amino acids. We determined the distribution of α6β4E within normal human glandular epithelium and its regulation and effect on cellular biophysical properties. Canonical α6β4C expressed in all basal cells, as expected, while α6β4E expressed within a subset of luminal cells. α6β4E expression was induced by three-dimensional culture conditions, activated Src, was reversible, and was stabilized by bortezomib, a proteasome inhibitor. α6β4C expressed in all cells during induced migration, whereas α6β4E was restricted to a subset of cells with increased kinetics of cell-cell and cell-ECM resistance properties. Interestingly, α6β4E presented in "ringlike" patterns measuring ∼1.75 × 0.72 microns and containing actin and CD9 at cell-ECM locations. In contrast, α6β4C expressed only within hemidesmosome-like structures containing BP180. Integrin α6β4E is an inducible adhesion isoform in normal epithelial cells that can alter biophysical properties of cell-cell and cell-ECM interactions.
• Cress, A. E., Nagle, R. B., Hinton, J. P., Wang, M., Gard, J. M., Garcia, J. G., & Knudsen, B. S. (2019). Integrin α6β4E variant is associated with actin and CD9 structures and modifies the biophysical properties of cell–cell and cell–extracellular matrix interactions. Molecular Biology of the Cell, 30(7), 838-850. doi:10.1091/mbc.e18-10-0652
• Sun, B., Moreno-vinasco, L., Valera, D. G., Valdez, M., Sun, B., Oita, R. C., Moreno-vinasco, L., Garcia, J. G., Garcia, A. N., Cress, A. E., & Carrie, K. (2019). NAMPT Is a Novel Participant and Therapeutic Target in Radiation-Induced Lung Injury (RILI). International Journal of Radiation Oncology Biology Physics, 105(1). doi:10.1164/ajrccm-conference.2019.199.1_meetingabstracts.a7255
• Wang, M., Rogers, G. C., & Cress, A. E. (2019). Immunofluorescence-based Determination of Centrosome Number in Tissue Samples.. Bio-protocol, 9(20), e3396. doi:10.21769/bioprotoc.3396
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  Centrosome numerical abnormalities have been reported in a variety of tumors. Centrosome numbers in cancer cells display both inter-tumor and intra-tumor heterogeneity. The over production of centrosomes (centrosome amplification) is unique in cancer cells and is a promising target for therapy. Thus, a method to quantify centrosome numbers on a single cell level is needed. Here, we describe a protocol to quantify centrosome numbers in formalin fixed paraffin embedded (FFPE) tissue samples by multiplexing antibodies to define bona fide centrosomes and cell borders. Centrosomes in single cells are identified using high resolution immunofluorescent microscopy with Z-sectioning. This protocol is easy to perform and has been used to quantify centrosome numbers on a single cell level in a variety of human tissue samples.
• Warfel, N. A., Nagle, R. B., Harryman, W. L., & Cress, A. E. (2019). The Tumor Microenvironments of Lethal Prostate Cancer.. Advances in experimental medicine and biology, 1210, 149-170. doi:10.1007/978-3-030-32656-2_8
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  Localized prostate cancer (confined to the gland) generally is considered curable, with nearly a 100% 5-year-survival rate. When the tumor escapes the prostate capsule, leading to metastasis, there is a poorer prognosis and higher mortality rate, with 5-year survival dropping to less than 30%. A major research question has been to understand the transition from indolent (low risk) disease to aggressive (high risk) disease. In this chapter, we provide details of the changing tumor microenvironments during prostate cancer invasion and their role in the progression and metastasis of lethal prostate cancer. Four microenvironments covered here include the muscle stroma, perineural invasion, hypoxia, and the role of microvesicles in altering the extracellular matrix environment. The adaptability of prostate cancer to these varied microenvironments and the cues for phenotypic changes are currently understudied areas. Model systems for understanding smooth muscle invasion both in vitro and in vivo are highlighted. Invasive human needle biopsy tissue and mouse xenograft tumors both contain smooth muscle invasion. In combination, the models can be used in an iterative process to validate molecular events for smooth muscle invasion in human tissue. Understanding the complex and interacting microenvironments in the prostate holds the key to early detection of high-risk disease and preventing tumor invasion through escape from the prostate capsule.
• Cress, A. E. (2018). Novel Regulation of Integrin Trafficking by Rab11-FIP5 in Aggressive Prostate Cancer. Molecular Cancer Research.
• Cress, A. E. (2018). Novel Regulation of Integrin Trafficking by Rab11-FIP5 in Aggressive Prostate Cancer.. Molecular cancer research : MCR.
• Cress, A. E. (2018). Regulation of inside-out β1-integrin activation by CDCP1.. Oncogene.
• Cress, A. E. (2018). Spatial Mapping of Myeloid Cells and Macrophages by Multiplexed Tissue Staining.. Frontiers in immunology.
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  An array of phenotypically diverse myeloid cells and macrophages (MC&M) resides in the tumor microenvironment, requiring multiplexed detection systems for visualization. Here we report an automated, multiplexed staining approach, named PLEXODY, that consists of five MC&M-related fluorescently-tagged antibodies (anti - CD68, - CD163, - CD206, - CD11b, and - CD11c), and three chromogenic antibodies, reactive with high- and low-molecular weight cytokeratins and CD3, highlighting tumor regions, benign glands and T cells. The staining prototype and image analysis methods which include a pixel/area-based quantification were developed using tissues from inflamed colon and tonsil and revealed a unique tissue-specific composition of 14 MC&M-associated pixel classes. As a proof-of-principle, PLEXODY was applied to three cases of pancreatic, prostate and renal cancers. Across digital images from these cancer types we observed 10 MC&M-associated pixel classes at frequencies greater than 3%. Cases revealed higher frequencies of single positive compared to multi-color pixels and a high abundance of CD68+/CD163+ and CD68+/CD163+/CD206+ pixels. Significantly more CD68+ and CD163+ vs. CD11b+ and CD11c+ pixels were in direct contact with tumor cells and T cells. While the greatest percentage (~70%) of CD68+ and CD163+ pixels was 0-20 microns away from tumor and T cell borders, CD11b+ and CD11c+ pixels were detected up to 240 microns away from tumor/T cell masks. Together, these data demonstrate significant differences in densities and spatial organization of MC&M-associated pixel classes, but surprising similarities between the three cancer types.
• Das, L., Gard, J. M., Prekeris, R., Nagle, R. B., Morrissey, C., Knudsen, B. S., Miranti, C. K., & Cress, A. E. (2018). Novel Regulation of Integrin Trafficking by Rab11-FIP5 in Aggressive Prostate Cancer. Molecular cancer research : MCR, 16(8), 1319-1331.
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  The laminin-binding integrins, α3β1 and α6β1, are needed for tumor metastasis and their surface expression is regulated by endocytic recycling. β1 integrins share the Rab11 recycling machinery, but the trafficking of α3β1 and α6β1 are distinct by an unknown mechanism. Using a mouse PDX tumor model containing human metastatic prostate cancer, Rab11 family interacting protein 5 (Rab11-FIP5) was identified as a lead candidate for α6β1 trafficking. Rab11-FIP5 and its membrane-binding domain were required for α6β1 recycling, without affecting the other laminin-binding integrin (i.e., α3β1) or unrelated membrane receptors like CD44, transferrin receptor, or E-cadherin. Depletion of Rab11-FIP5 resulted in the intracellular accumulation of α6β1 in the Rab11 recycling compartment, loss of cell migration on laminin, and an unexpected loss of α6β1 recycling in cell-cell locations. Taken together, these data demonstrate that α6β1 is distinct from α3β1 via Rab11-FIP5 recycling and recycles in an unexpected cell-cell location. Rab11-FIP5-dependent α6β1 integrin recycling may be selectively targeted to limit migration of prostate cancer cells into laminin-rich tissues. .
• Das, L., Gard, J., Prekeris, R., Nagle, R. B., Morrissey, C., Knudsen, B. S., Miranti, C. K., & Cress, A. E. (2018). Novel Regulation of Integrin Trafficking by Rab11-FIP5 in Aggressive Prostate Cancer. MOLECULAR CANCER RESEARCH, 16(8), 1319-1331.
• Epshtein, Y., Anis, M., Chen, W., Wang, T., Cress, A. E., & Jacobson, J. R. (2018). SYNDECAN-1 MEDIATES LUNG ENDOTHELIAL CELL INFLAMMATORY SIGNALING BY INTEGRIN beta 4. JOURNAL OF INVESTIGATIVE MEDICINE, 66(4), 872-872.
• Pollan, S. G., Huang, F., Sperger, J. M., Lang, J. M., Morrissey, C., Cress, A. E., Chu, C. Y., Bhowmick, N. A., You, S., Freeman, M. R., Spassov, D. S., Moasser, M. M., Carter, W. G., Satapathy, S. R., Shah, K., & Knudsen, B. S. (2018). Regulation of inside-out beta 1-integrin activation by CDCP1. ONCOGENE, 37(21), 2817-2836.
• Pollan, S. G., Huang, F., Sperger, J. M., Lang, J. M., Morrissey, C., Cress, A. E., Chu, C. Y., Bhowmick, N. A., You, S., Freeman, M. R., Spassov, D. S., Moasser, M. M., Carter, W. G., Satapathy, S. R., Shah, K., & Knudsen, B. S. (2018). Regulation of inside-out β1-integrin activation by CDCP1. Oncogene, 37(21), 2817-2836.
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  Tumor metastasis depends on the dynamic regulation of cell adhesion through β1-integrin. The Cub-Domain Containing Protein-1, CDCP1, is a transmembrane glycoprotein which regulates cell adhesion. Overexpression and loss of CDCP1 have been observed in the same cancer types to promote metastatic progression. Here, we demonstrate reduced CDCP1 expression in high-grade, primary prostate cancers, circulating tumor cells and tumor metastases of patients with castrate-resistant prostate cancer. CDCP1 is expressed in epithelial and not mesenchymal cells, and its cell surface and mRNA expression declines upon stimulation with TGFβ1 and epithelial-to-mesenchymal transition. Silencing of CDCP1 in DU145 and PC3 cells resulted in 3.4-fold higher proliferation of non-adherent cells and 4.4-fold greater anchorage independent growth. CDCP1-silenced tumors grew in 100% of mice, compared to 30% growth of CDCP1-expressing tumors. After CDCP1 silencing, cell adhesion and migration diminished 2.1-fold, caused by loss of inside-out activation of β1-integrin. We determined that the loss of CDCP1 reduces CDK5 kinase activity due to the phosphorylation of its regulatory subunit, CDK5R1/p35, by c-SRC on Y234. This generates a binding site for the C2 domain of PKCδ, which in turn phosphorylates CDK5 on T77. The resulting dissociation of the CDK5R1/CDK5 complex abolishes the activity of CDK5. Mutations of CDK5-T77 and CDK5R1-Y234 phosphorylation sites re-establish the CDK5/CDKR1 complex and the inside-out activity of β1-integrin. Altogether, we discovered a new mechanism of regulation of CDK5 through loss of CDCP1, which dynamically regulates β1-integrin in non-adherent cells and which may promote vascular dissemination in patients with advanced prostate cancer.
• Pollan, S., Sperger, J., Lang, J., Huang, F., Zheng, J., Morrissey, C., Cress, A. E., Spassov, D., Moasser, M., Carter, W., & Knudsen, B. (2018). CDCP1, a potential noninvasive biomarker in circulating tumor cells for treatment of prostate cancer with FAK inhibitors.. CANCER RESEARCH, 78(16), 131-132.
• Saylor, J., Ma, Z., Goodridge, H. S., Huang, F., Cress, A. E., Pandol, S. J., Shiao, S. L., Vidal, A. C., Wu, L., Nickols, N. G., Gertych, A., & Knudsen, B. S. (2018). Spatial Mapping of Myeloid Cells and Macrophages by Multiplexed Tissue Staining. FRONTIERS IN IMMUNOLOGY, 9.
• Saylor, J., Weterings, E., Ellis, N., Hinton, J., Roberts, E., Cress, A., & Knudsen, B. (2018). An automated tissue staining and quantitative digital image analysis pipeline for quantification of DNA damage repair at the single-cell level.. CANCER RESEARCH, 78(16), 130-131.
• Schmelz, M., Cress, A. E., Scott, K. M., Bürger, F., Cui, H., Sallam, K., McDaniel, K. M., Dalkin, B. L., & Nagle, R. B. (2018). Different phenotypes in human prostate cancer: alpha6 or alpha3 integrin in cell-extracellular adhesion sites. Neoplasia (New York, N.Y.), 4(3), 243-54.
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  The distribution of alpha6/alpha3 integrin in adhesion complexes at the basal membrane in human normal and cancer prostate glands was analyzed in 135 biopsies from 61 patients. The levels of the polarized alpha6/alpha3 integrin expression at the basal membrane of prostate tumor glands were determined by quantitative immunohistochemistry. The alpha6/alpha3 integrin expression was compared with Gleason sum score, pathological stage, and preoperative serum prostate-specific antigen (PSA). The associations were assessed by statistical methods. Eighty percent of the tumors expressed the alpha6 or alpha3 integrin and 20% was integrin-negative. Gleason sum score, but not serum PSA, was associated with the integrin expression. Low Gleason sum score correlated with increased integrin expression, high Gleason sum score with low and negative integrin expression. Three prostate tumor phenotypes were distinguished based on differential integrin expression. Type I coexpressed both alpha6 and alpha3 subunits, type II exclusively expressed alpha6 integrin, and type III expressed alpha3 integrin only. Fifteen cases were further examined for the codistribution of vinculin, paxillin, and CD 151 on frozen serial sections using confocal laser scanning microscopy. The alpha6/alpha3 integrins, CD151, paxillin, and vinculin were present within normal glands. In prostate carcinoma, alpha6 integrin was colocalized with CD 151, but not with vinculin or paxillin. In tumor phenotype I, the alpha6 subunit did not colocalize with the alpha subunit indicating the existence of two different adhesion complexes. Human prostate tumors display on their cell surface the alpha6beta1 and/or alpha3beta1 integrins. Three tumor phenotypes associated with two different adhesion complexes were identified, suggesting a reorganization of cell adhesion structures in prostate cancer.
• Wang, M., Knudsen, B. S., Rogers, G. C., & Cress, A. E. (2018). Centrosome loss and chromosomal instability in prostate tumor progression.. CANCER RESEARCH, 78(16), 50-51.
• Wang, M., Rogers, G. C., Knudsen, B. S., & Cress, A. E. (2018). Abstract A049: Centrosome loss and chromosomal instability in prostate tumor progression. Cancer Research, 78. doi:10.1158/1538-7445.prca2017-a049
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  Background: Chromosomal instability is a hallmark of cancer. Early genomic events in tumor progression include copy number variations such as tandem duplications and deletions that involve at least >100kB stretches of DNA. Although single-driver mutations are remarkably infrequent in most cancers (e.g., accounting for only 2-3% of human epithelial cancers), approximately 20% of prostate tumors are reported to display chromothripsis. Errors in mitotic chromosome segregation produce micronuclei that are susceptible to chromothripsis, and this can occur due to changes in centrosome numbers. Centrosomes are organelles that nucleate microtubule growth and, thus, determine the number of mitotic spindle poles. Normally, mitotic cells contain two centrosomes that guide formation of a bipolar spindle, ensuring that daughter cells divide within the epithelial plane and inherit an equal complement of the genome. Centrioles are the duplicating elements of centrosomes, and their loss or overduplication (known as “amplification”) promotes spindle assembly defects that lead to chromothripsis. Precise control of centriole copy number is vital in order to maintain genomic stability and normal tissue homeostasis. Methods: We developed an assay for the detection and quantitation of centrosome numbers in FFPE normal and cancer tissue and tissue culture cell lines. In addition, a 3D culture model of an immortalized nontumorigenic prostate epithelial cell line (RWPE-1) was used to characterize the progression of prostate spheroids from normal gland to high-grade prostatic intraepithelial neoplasia (HG-PIN) type structures in vitro. Genomic instability was detected by polyploidy (flow and metaphase analysis), SKY (Roswell Park Cancer Institute Cytogenetics SKY core laboratory), and micronuclei formation. Chromosome fragility was assessed by time-lapse microscopy. Results: Human prostate cancer has a readily detectable centrosome dysfunction that occurs early in the disease progression. Whereas triple-negative breast cancer specimens contained centrosome amplification as expected, surprisingly, centrosome loss was detected in early-stage prostate cancer (Gleason stage 3+3, 3+4 and 4+4) tissue specimens and in AR-positive cell lines (LnCAP, VCaP). We note that centrosome amplification was observed in AR-negative cell lines (DU145, PC3 and H660). Using TCGA data, twelve centriolar genes were either amplified, deleted, or mutated in prostate adenocarcinoma and metastatic disease. In contrast, all twelve centriolar genes were highly amplified in neuroendocrine prostate cancer. Using centrinone, a small-molecule inhibitor of Polo-like kinase 4, we blocked centriole duplication in nontumorigenic prostate cells (RWPE-1). Centrinone treatment resulted in significant genomic instability (polyploidy, copy number variation across all chromosomes, and micronuclei formation) and mitotic errors (increased mitotic index, prolonged mitosis, and increased chromosome fragility). Additionally, centrosome loss produced prostate cancer-specific phenotypes (erg overexpression, TMPRSS2 amplification, and invasive budding). Conclusions: The loss of centrosomes during early stages of prostate cancer development may account for the increased chromosomal instability manifested as polyploidy and copy number variations. Whether alterations in centrosome numbers occur in high-grade prostate cancer or metastatic lesions is unknown. Centrosomes may be promising biomarkers for advancing disease but also rational targets for personalized therapy in prostate cancer. (Supported in part by NIH grants CA 23074, CA 159406 and NIGMS R01GFM110166.) Citation Format: Mengdie Wang, Beatrice S. Knudsen, Gregory C. Rogers, Anne E. Cress. Centrosome loss and chromosomal instability in prostate tumor progression [abstract]. In: Proceedings of the AACR Special Conference: Prostate Cancer: Advances in Basic, Translational, and Clinical Research; 2017 Dec 2-5; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(16 Suppl):Abstract nr A049.
• Cress, A. E. (2017). Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells. Journal of Cellular Biochemistry.
• Cress, A. E. (2017). Targeting the Cohesive Cluster Phenotype in Chordoma via β1 Integrin Increases Ionizing Radiation Efficacy.. Neoplasia (New York, N.Y.).
• Cress, A. E., Nagle, R. B., Das, L., Anderson, T. A., Gard, J. M., Sroka, I. C., Strautman, S. R., Morrissey, C., & Knudsen, B. S. (2017). Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells: LAMININBINDINGINTEGRININTERNALIZATION. Journal of Cellular Biochemistry, 118(5), 1038-1049. doi:10.1002/jcb.25673
• Cress, A. E., Nagle, R. B., Das, L., Anderson, T. A., Gard, J. M., Sroka, I. C., Strautman, S. R., Morrissey, C., & Knudsen, B. S. (2017). Cover Image, Volume 118, Number 5, May 2017. Journal of Cellular Biochemistry, 118(5), i-i. doi:10.1002/jcb.25989
• Das, L., Anderson, T. A., Gard, J. M., Sroka, I. C., Strautman, S. R., Nagle, R. B., Morrissey, C., Knudsen, B. S., & Cress, A. E. (2017). Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells. Journal of cellular biochemistry, 118(5), 1038-1049.
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  Laminin binding integrins α6 (CD49f) and α3 (CD49c) are persistently but differentially expressed in prostate cancer (PCa). Integrin internalization is an important determinant of their cell surface expression and function. Using flow cytometry, and first order kinetic modeling, we quantitated the intrinsic internalization rates of integrin subunits in a single cycle of internalization. In PCa cell line DU145, α6 integrin internalized with a rate constant (k ) of 3.25 min , threefold faster than α3 integrin (1.0 min ), 1.5-fold faster than the vitronectin binding αv integrin (CD51) (2.2 min ), and significantly slower than the unrelated transferrin receptor (CD71) (15 min ). Silencing of α3 integrin protein expression in DU145, PC3, and PC3B1 cells resulted in up to a 1.71-fold increase in k for α6 integrin. The internalized α6 integrin was targeted to early endosomes but not to lamp1 vesicles. Depletion of α3 integrin expression resulted in redistribution of α6β4 integrin to an observed cell-cell staining pattern that is consistent with a suprabasal distribution observed in epidermis and early PIN lesions in PCa. Depletion of α3 integrin increased cell migration by 1.8-fold, which was dependent on α6β1 integrin. Silencing of α6 integrin expression however, had no significant effect on the k of α3 integrin or its distribution in early endosomes. These results indicate that α3 and α6 integrins have significantly different internalization kinetics and that coordination exists between them for internalization. J. Cell. Biochem. 118: 1038-1049, 2017. © 2016 Wiley Periodicals, Inc.
• Das, L., Anderson, T. A., Gard, J., Sroka, I. C., Strautman, S. R., Nagle, R. B., Morrissey, C., Knudsen, B. S., & Cress, A. E. (2017). Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells. JOURNAL OF CELLULAR BIOCHEMISTRY, 118(5), 1038-1049.
• Harryman, W. L., Gard, J. M., Pond, K. W., Simpson, S. J., Heppner, L. H., Hernandez-Cortes, D., Little, A. S., Eschbacher, J. M., & Cress, A. E. (2017). Targeting the Cohesive Cluster Phenotype in Chordoma via β1 Integrin Increases Ionizing Radiation Efficacy. Neoplasia (New York, N.Y.), 19(11), 919-927.
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  Chordoma is a rare, radiation-resistant, skull-base and spinal tumor with high local recurrence containing mixed cell-adhesion phenotypes. We characterized DNA damage response (DDR) signaling (γH2AX, pKAP1, pATM) and survival response to ionizing radiation (IR) in human chordoma samples (42 resections, 23 patients) to test if blocking cell adhesion sensitizes U-CH1 tumor cells to IR. U-CH1 cells expressed brachyury, YAP, and laminin adhesion receptors (CD49c, CD49f, CD44), and approximately 15% to 20% of U-CH1 cells featured an α6 integrin-dependent (CD49f) cohesive cluster phenotype, which confers therapeutic resistance and aids metastasis. DDR to IR in U-CH1 cells was compared to normal prostate epithelial (PrEC) and tumor cells (DU145). Flow cytometry showed a dose- and time-dependent increase in γH2AX and pKAP1 expression in all cell lines. However, nearly 50% of U-CH1 cells exhibited nonresponsive phenotype to IR (measured by γH2AX and pKAP1) independent of cell cycle status. Immunofluorescence microscopy verified that only 15% of U-CH1 clustered cells were γH2AX or pKAP1 positive (versus 80% of nonclustered cells) 2 hours following 2-Gy IR. Conversely, both tumor cell lines were uniformly defective in pATM response. HYD1, a synthetic ECM ligand, inhibited DDR through an unresolved γH2AX response. β1 integrin-blocking antibody (AIIB2) decreased cell survival 50% itself and approximately doubled the IR-induced cell kill at all IR doses observed at 2 and 4 weeks posttreatment. These results suggest that a heterogeneity of DDR to IR exists within a chordoma population. Blocking integrin function alone and/or as an adjuvant to IR may eradicate chordomas containing the cohesive cluster phenotype.
• Harryman, W. L., Gard, J., Pond, K. W., Simpson, S. J., Heppner, L. H., Hernandez-Cortes, D., Little, A. S., Eschbacher, J. M., & Cress, A. E. (2017). Targeting the Cohesive Cluster Phenotype in Chordoma via beta 1 Integrin Increases Ionizing Radiation Efficacy. NEOPLASIA, 19(11), 919-927.
• Wang, M., Nagle, R. B., Knudsen, B. S., Rogers, G. C., & Cress, A. E. (2017). A basal cell defect promotes budding of prostatic intraepithelial neoplasia. JOURNAL OF CELL SCIENCE, 130(1), 104-110.
• Wang, M., Nagle, R. B., Knudsen, B. S., Rogers, G. C., & Cress, A. E. (2017). A basal cell defect promotes budding of prostatic intraepithelial neoplasia. Journal of cell science, 130(1), 104-110.
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  Basal cells in a simple secretory epithelium adhere to the extracellular matrix (ECM), providing contextual cues for ordered repopulation of the luminal cell layer. Early high-grade prostatic intraepithelial neoplasia (HG-PIN) tissue has enlarged nuclei and nucleoli, luminal layer expansion and genomic instability. Additional HG-PIN markers include loss of α6β4 integrin or its ligand laminin-332, and budding of tumor clusters into laminin-511-rich stroma. We modeled the invasive budding phenotype by reducing expression of α6β4 integrin in spheroids formed from two normal human stable isogenic prostate epithelial cell lines (RWPE-1 and PrEC 11220). These normal cells continuously spun in culture, forming multicellular spheroids containing an outer laminin-332 layer, basal cells (expressing α6β4 integrin, high-molecular-weight cytokeratin and p63, also known as TP63) and luminal cells that secrete PSA (also known as KLK3). Basal cells were optimally positioned relative to the laminin-332 layer as determined by spindle orientation. β4-integrin-defective spheroids contained a discontinuous laminin-332 layer corresponding to regions of abnormal budding. This 3D model can be readily used to study mechanisms that disrupt laminin-332 continuity, for example, defects in the essential adhesion receptor (β4 integrin), laminin-332 or abnormal luminal expansion during HG-PIN progression.
• Cress, A. E. (2016). A basal cell defect promotes budding of prostatic intraepithelial neoplasia. Journal of Cell Science.
• Cress, A. E. (2016). Laminin-binding integrin gene copy number alterations in distinct epithelial-type cancers.. American journal of translational research.
• Cress, A. E. (2016). The Cohesive Metastasis Phenotype in Human Prostate Cancer.. Biochimica et biophysica acta.
• Cress, A. E., Rogers, G. C., Wang, M., Nagle, R. B., & Knudsen, B. S. (2016). A basal cell defect promotes budding of prostatic intraepithelial neoplasia. Journal of Cell Science. doi:10.1242/jcs.188177
• Das, L., Anderson, T. A., Gard, J. M., Sroka, I. C., Strautman, S. R., Nagle, R. B., Morrissey, C., Knudsen, B. S., & Cress, A. E. (2016). Loss of alpha 3 integrin expression promotes a6 integrin internalization to Rab4 vesicles and migration of human prostate cancer cells. CANCER RESEARCH, 76.
• Harryman, W. L., Hinton, J. P., Rubenstein, C. P., Singh, P., Nagle, R. B., Parker, S. J., Knudsen, B. S., & Cress, A. E. (2016). The Cohesive Metastasis Phenotype in Human Prostate Cancer. BIOCHIMICA ET BIOPHYSICA ACTA-REVIEWS ON CANCER, 1866(2), 221-231.
• Harryman, W. L., Hinton, J. P., Rubenstein, C. P., Singh, P., Nagle, R. B., Parker, S. J., Knudsen, B. S., & Cress, A. E. (2016). The Cohesive Metastasis Phenotype in Human Prostate Cancer. Biochimica et biophysica acta, 1866(2), 221-231.
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  A critical barrier for the successful prevention and treatment of recurrent prostate cancer is detection and eradication of metastatic and therapy-resistant disease. Despite the fall in diagnoses and mortality, the reported incidence of metastatic disease has increased 72% since 2004. Prostate cancer arises in cohesive groups as intraepithelial neoplasia, migrates through muscle and leaves the gland via perineural invasion for hematogenous dissemination. Current technological advances have shown cohesive-clusters of tumor (also known as microemboli) within the circulation. Circulating tumor cell (CTC) profiles are indicative of disseminated prostate cancer, and disseminated tumor cells (DTC) are found in cohesive-clusters, a phenotypic characteristic of both radiation- and drug-resistant tumors. Recent reports in cell biology and informatics, coupled with mass spectrometry, indicate that the integrin adhesome network provides an explanation for the biophysical ability of cohesive-clusters of tumor cells to invade thorough muscle and nerve microenvironments while maintaining adhesion-dependent therapeutic resistance. Targeting cohesive-clusters takes advantage of the known ability of extracellular matrix (ECM) adhesion to promote tumor cell survival and represents an approach that has the potential to avoid the progression to drug- and radiotherapy-resistance. In the following review we will examine the evidence for development and dissemination of cohesive-clusters in metastatic prostate cancer.
• Harryman, W. L., Pond, E., Singh, P., Little, A. S., Eschbacher, J. M., Nagle, R. B., & Cress, A. E. (2016). Laminin-binding integrin gene copy number alterations in distinct epithelial-type cancers. American journal of translational research, 8(2), 940-54.
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  The laminin-binding integrin (LBI) family are cell adhesion molecules that are essential for invasion and metastasis of human epithelial cancers and cell adhesion mediated drug resistance. We investigated whether copy number alteration (CNA) or mutations of a five-gene signature (ITGB4, ITGA3, LAMB3, PLEC, and SYNE3), representing essential genes for LBI adhesion, would correlate with patient outcomes within human epithelial-type tumor data sets currently available in an open access format.
• Pond, E., Harryman, W. L., & Cress, A. E. (2016). Laminin binding integrin copy number variations in distinct types of epithelial cancers. CLINICAL CANCER RESEARCH, 22.
• Song, S., Jacobson, K. N., McDermott, K. M., Reddy, S. P., Cress, A. E., Tang, H., Dudek, S. M., Black, S. M., Garcia, J. G., Makino, A., & Yuan, J. X. (2016). ATP promotes cell survival via regulation of cytosolic [Ca2+] and Bcl-2/Bax ratio in lung cancer cells. American journal of physiology. Cell physiology, 310(2), C99-114.
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  Adenosine triphosphate (ATP) is a ubiquitous extracellular messenger elevated in the tumor microenvironment. ATP regulates cell functions by acting on purinergic receptors (P2X and P2Y) and activating a series of intracellular signaling pathways. We examined ATP-induced Ca(2+) signaling and its effects on antiapoptotic (Bcl-2) and proapoptotic (Bax) proteins in normal human airway epithelial cells and lung cancer cells. Lung cancer cells exhibited two phases (transient and plateau phases) of increase in cytosolic [Ca(2+)] ([Ca(2+)]cyt) caused by ATP, while only the transient phase was observed in normal cells. Removal of extracellular Ca(2+) eliminated the plateau phase increase of [Ca(2+)]cyt in lung cancer cells, indicating that the plateau phase of [Ca(2+)]cyt increase is due to Ca(2+) influx. The distribution of P2X (P2X1-7) and P2Y (P2Y1, P2Y2, P2Y4, P2Y6, P2Y11) receptors was different between lung cancer cells and normal cells. Proapoptotic P2X7 was nearly undetectable in lung cancer cells, which may explain why lung cancer cells showed decreased cytotoxicity when treated with high concentration of ATP. The Bcl-2/Bax ratio was increased in lung cancer cells following treatment with ATP; however, the antiapoptotic protein Bcl-2 demonstrated more sensitivity to ATP than proapoptotic protein Bax. Decreasing extracellular Ca(2+) or chelating intracellular Ca(2+) with BAPTA-AM significantly inhibited ATP-induced increase in Bcl-2/Bax ratio, indicating that a rise in [Ca(2+)]cyt through Ca(2+) influx is the critical mediator for ATP-mediated increase in Bcl-2/Bax ratio. Therefore, despite high ATP levels in the tumor microenvironment, which would induce cell apoptosis in normal cells, the decreased P2X7 and elevated Bcl-2/Bax ratio in lung cancer cells may enable tumor cells to survive. Increasing the Bcl-2/Bax ratio by exposure to high extracellular ATP may, therefore, be an important selective pressure promoting transformation and cancer progression.
• Song, S., Jacobson, K. N., McDermott, K. M., Reddy, S. P., Cress, A. E., Tang, H., Dudek, S. M., Black, S. M., Garcia, J., Makino, A., & Yuan, J. (2016). ATP promotes cell survival via regulation of cytosolic [Ca2+] and Bcl-2/Bax ratio in lung cancer cells. AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 310(2), C99-C114.
• Sroka, I. C., Chopra, H., Das, L., Gard, J. M., Nagle, R. B., & Cress, A. E. (2016). Schwann Cells Increase Prostate and Pancreatic Tumor Cell Invasion Using Laminin Binding A6 Integrin. Journal of cellular biochemistry, 117(2), 491-9.
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  Human pancreatic and prostate cancers metastasize along nerve axons during perineural invasion. The extracellular matrix laminin class of proteins is an abundant component of both myelinated and non-myelinated nerves. Analysis of human pancreatic and prostate tissue revealed both perineural and endoneural invasion with Schwann cells surrounded or disrupted by tumor, respectively. Tumor and nerve cell co-culture conditions were used to determine if myelinating or non-myelinating Schwann cell (S16 and S16Y, respectively) phenotype was equally likely to promote integrin-dependent cancer cell invasion and migration on laminin. Conditioned medium from S16 cells increased tumor cell (DU145, PC3, and CFPAC1) invasion into laminin approximately 1.3-2.0 fold compared to fetal bovine serum (FBS) treated cells. Integrin function (e.g., ITGA6p formation) increased up to 1.5 fold in prostate (DU145, PC3, RWPE-1) and pancreatic (CFPAC1) cells, and invasion was dependent on ITGA6p formation and ITGB1 as determined by function-blocking antibodies. In contrast, conditioned medium isolated from S16Y cells (non-myelinating phenotype) decreased constitutive levels of ITGA6p in the tumor cells by 50% compared to untreated cells and decreased ITGA6p formation 3.0 fold compared to S16 treated cells. Flow cytometry and western blot analysis revealed loss of ITGA6p formation as reversible and independent of overall loss of ITGA6 expression. These results suggest that the myelinating phenotype of Schwann cells within the tumor microenvironment increased integrin-dependent tumor invasion on laminin.
• Sroka, I. C., Chopra, H., Das, L., Gard, J., Nagle, R. B., & Cress, A. E. (2016). Schwann Cells Increase Prostate and Pancreatic Tumor Cell Invasion Using Laminin Binding A6 Integrin. JOURNAL OF CELLULAR BIOCHEMISTRY, 117(2), 491-499.
• Algotar, A. M., Cress, A., Nagle, R., Sokoloff, M., Behnejad, R., Drinkwitz, D., Lodhia, N., Thompson, P., & Stratton, S. P. (2015). DNA damage and repair markers to improve accuracy of prostate cancer diagnosis and identification of aggressive disease.. JOURNAL OF CLINICAL ONCOLOGY, 33(15).
• Algotar, A., Cress, A., Nagle, R., Sokoloff, M., Singh, P., Behnejad, R., & Stratton, S. (2015). Association of ERG-PTEN expression profile of prostate biopsy with PSA velocity.. JOURNAL OF CLINICAL ONCOLOGY, 33(7).
• Chen, W., Epshtein, Y., Ni, X., Dull, R. O., Cress, A. E., Garcia, J. G., & Jacobson, J. R. (2015). Role of Integrin β4 in Lung Endothelial Cell Inflammatory Responses to Mechanical Stress. Scientific reports, 5, 16529.
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  Simvastatin, an HMG-CoA reductase inhibitor, has lung vascular-protective effects that are associated with decreased agonist-induced integrin β4 (ITGB4) tyrosine phosphorylation. Accordingly, we hypothesized that endothelial cell (EC) protection by simvastatin is dependent on these effects and sought to further characterize the functional role of ITGB4 as a mediator of EC protection in the setting of excessive mechanical stretch at levels relevant to ventilator-induced lung injury (VILI). Initially, early ITGB4 tyrosine phosphorylation was confirmed in human pulmonary artery EC subjected to excessive cyclic stretch (18% CS). EC overexpression of mutant ITGB4 with specific tyrosines mutated to phenylalanine (Y1440, Y1526 Y1640, or Y1422) resulted in significantly attenuated CS-induced cytokine expression (IL6, IL-8, MCP-1, and RANTES). In addition, EC overexpression of ITGB4 constructs with specific structural deletions also resulted in significantly attenuated CS-induced inflammatory cytokine expression compared to overexpression of wildtype ITGB4. Finally, mice expressing a mutant ITGB4 lacking a cytoplasmic signaling domain were found to have attenuated lung injury after VILI-challenge (VT = 40 ml/kg, 4 h). Our results provide mechanistic insights into the anti-inflammatory properties of statins and may ultimately lead to novel strategies targeted at ITGB4 signaling to treat VILI.
• Chen, W., Epshtein, Y., Ni, X., Dull, R. O., Cress, A. E., Garcia, J., & Jacobson, J. R. (2015). Role of Integrin beta 4 in Lung Endothelial Cell Inflammatory Responses to Mechanical Stress. SCIENTIFIC REPORTS, 5.
• Cress, A. E. (2015). ATP promotes cell survival via regulation of cytosolic [Ca2+] and Bcl-2/Bax ratio in lung cancer cells.. American journal of physiology. Cell physiology.
• Cress, A. E. (2015). Combined micro CT and histopathology for evaluation of skeletal metastasis in live animals.. American journal of translational research.
• Cress, A. E. (2015). Gemcitabine resistant pancreatic cancer cell lines acquire an invasive phenotype with collateral hypersensitivity to histone deacetylase inhibitors.. Cancer biology & therapy.
• Cress, A. E. (2015). Nuclear factor, erythroid 2-like 2-associated molecular signature predicts lung cancer survival.. Scientific reports.
• Cress, A. E. (2015). Role of Integrin β4 in Lung Endothelial Cell Inflammatory Responses to Mechanical Stress.. Scientific reports.
• Cress, A. E. (2015). Role played by paxillin and paxillin tyrosine phosphorylation in hepatocyte growth factor/sphingosine-1-phosphate-mediated reactive oxygen species generation, lamellipodia formation, and endothelial barrier function.. Pulmonary circulation.
• Cress, A. E. (2015). Schwann Cells Increase Prostate and Pancreatic Tumor Cell Invasion Using Laminin Binding A6 Integrin. Journal of Cellular Biochemistry.
• Fu, P., Usatyuk, P. V., Jacobson, J., Cress, A. E., Garcia, J. G., Salgia, R., & Natarajan, V. (2015). Role played by paxillin and paxillin tyrosine phosphorylation in hepatocyte growth factor/sphingosine-1-phosphate-mediated reactive oxygen species generation, lamellipodia formation, and endothelial barrier function. Pulmonary circulation, 5(4), 619-30.
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  Paxillin is a multifunctional and multidomain focal adhesion adaptor protein. It serves as an important scaffolding protein at focal adhesions by recruiting and binding to structural and signaling molecules. Paxillin tyrosine phosphorylation at Y31 and Y118 is important for paxillin redistribution to focal adhesions and angiogenesis. Hepatocyte growth factor (HGF) and sphingosine-1-phosphate (S1P) are potent stimulators of lamellipodia formation, a prerequisite for endothelial cell migration. The role played by paxillin and its tyrosine phosphorylated forms in HGF- or S1P-induced lamellipodia formation and barrier function is unclear. HGF or S1P stimulated lamellipodia formation, tyrosine phosphorylation of paxillin at Y31 and Y118, and c-Abl in human lung microvascular endothelial cells (HLMVECs). Knockdown of paxillin with small interfering RNA (siRNA) or transfection with paxillin mutants (Y31F or Y118F) mitigated HGF- or S1P-induced lamellipodia formation, translocation of p47 (phox) to lamellipodia, and reactive oxygen species (ROS) generation in HLMVECs. Furthermore, exposure of HLMVECs to HGF or S1P stimulated c-Abl-mediated tyrosine phosphorylation of paxillin at Y31 and Y118 in a time-dependent fashion, and down-regulation of c-Abl with siRNA attenuated HGF- or S1P-mediated lamellipodia formation, translocation of p47 (phox) to lamellipodia, and endothelial barrier enhancement. In vivo, knockdown of paxillin with siRNA in mouse lungs attenuated ventilator-induced lung injury. Together, these results suggest that c-Abl-mediated tyrosine phosphorylation of paxillin at Y31 and Y118 regulates HGF- or S1P-mediated lamellipodia formation, ROS generation in lamellipodia, and endothelial permeability.
• Fu, P., Usatyuk, P. V., Jacobson, J., Cress, A. E., Garcia, J., Salgia, R., & Natarajan, V. (2015). Role played by paxillin and paxillin tyrosine phosphorylation in hepatocyte growth factor/sphingosine-1-phosphate-mediated reactive oxygen species generation, lamellipodia formation, and endothelial barrier function. PULMONARY CIRCULATION, 5(4), 619-630.
• Geffre, C. P., Pond, E., Pond, G. D., Sroka, I. C., Gard, J. M., Skovan, B. A., Meek, W. E., Landowski, T. H., Nagle, R. B., & Cress, A. E. (2015). Combined micro CT and histopathology for evaluation of skeletal metastasis in live animals. AMERICAN JOURNAL OF TRANSLATIONAL RESEARCH, 7(2), 348-355.
• Geffre, C. P., Pond, E., Pond, G. D., Sroka, I. C., Gard, J. M., Skovan, B. A., Meek, W. E., Landowski, T. H., Nagle, R. B., & Cress, A. E. (2015). Combined micro CT and histopathology for evaluation of skeletal metastasis in live animals. American journal of translational research, 7(2), 348-55.
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  Bone is a favored site for solid tumor metastasis, especially among patients with breast, lung or prostate carcinomas. Micro CT is a powerful and inexpensive tool that can be used to investigate tumor progression in xenograft models of human disease. Many previous studies have relied on terminal analysis of harvested bones to document metastatic tumor activity. The current protocol uses live animals and combines sequential micro CT evaluation of lesion development with matched histopathology at the end of the study. The approach allows for both rapid detection and evaluation of bone lesion progression in live animals. Bone resident tumors are established either by direct (intraosseous) or arterial (intracardiac) injection, and lesion development is evaluated for up to eight weeks. This protocol provides a clinically relevant method for investigating bone metastasis progression and the development of osteotropic therapeutic strategies for the treatment of bone metastases.
• Qian, Z., Zhou, T., Gurguis, C. I., Xu, X., Wen, Q., Lv, J., Fang, F., Hecker, L., Cress, A. E., Natarajan, V., Jacobson, J. R., Zhang, D. D., Garcia, J. G., & Wang, T. (2015). Nuclear factor, erythroid 2-like 2-associated molecular signature predicts lung cancer survival. Scientific reports, 5, 16889.
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  Nuclear factor, erythroid 2-like 2 (NFE2L2), a transcription factor also known as NF-E2-related factor 2 (Nrf2), is a key cytoprotective gene that regulates critical antioxidant and stress-responsive genes. Nrf2 has been demonstrated to be a promising therapeutic target and useful biomarker in malignant disease. We hypothesized that NFE2L2-mediated gene expression would reflect cancer severity and progression. We conducted a meta-analysis of microarray data for 240 NFE2L2-mediated genes that were enriched in tumor tissues. We then developed a risk scoring system based on NFE2L2 gene expression profiling and designated 50 tumor-associated genes as the NFE2L2-associated molecular signature (NAMS). We tested the relationship between this gene expression signature and both recurrence-free survival and overall survival in lung cancer patients. We find that NAMS predicts clinical outcome in the training cohort and in 12 out of 20 validation cohorts. Cox proportional hazard regressions indicate that NAMS is a robust prognostic gene signature, independent of other clinical and pathological factors including patient age, gender, smoking, gene alteration, MYC level, and cancer stage. NAMS is an excellent predictor of recurrence-free survival and overall survival in human lung cancer. This gene signature represents a promising prognostic biomarker in human lung cancer.
• Qian, Z., Zhou, T., Gurguis, C. I., Xu, X., Wen, Q., Lv, J., Fang, F., Hecker, L., Cress, A. E., Natarajan, V., Jacobson, J. R., Zhang, D. D., Garcia, J., & Wang, T. (2015). Nuclear factor, erythroid 2-like 2-associated molecular signature predicts lung cancer survival. SCIENTIFIC REPORTS, 5.
• Samulitis, B. K., Pond, E., Pond, K., Cress, A. E., Dorr, R. T., & Landowski, T. H. (2015). Adaptive drug resistance is associated with phenotypic reprogramming and hypersensitivity to HDAC inhibitors. CLINICAL CANCER RESEARCH, 21.
• Samulitis, B. K., Pond, K. W., Pond, E., Cress, A. E., Patel, H., Wisner, L., Patel, C., Dorr, R. T., & Landowski, T. H. (2015). Gemcitabine resistant pancreatic cancer cell lines acquire an invasive phenotype with collateral hypersensitivity to histone deacetylase inhibitors. CANCER BIOLOGY & THERAPY, 16(1), 43-51.
• Samulitis, B. K., Pond, K. W., Pond, E., Cress, A. E., Patel, H., Wisner, L., Patel, C., Dorr, R. T., & Landowski, T. H. (2015). Gemcitabine resistant pancreatic cancer cell lines acquire an invasive phenotype with collateral hypersensitivity to histone deacetylase inhibitors. Cancer biology & therapy, 16(1), 43-51.
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  Gemcitabine based treatment is currently a standard first line treatment for patients with advanced pancreatic cancer, however overall survival remains poor, and few options are available for patients that fail gemcitabine based therapy. To identify potential molecular targets in gemcitabine refractory pancreatic cancer, we developed a series of gemcitabine resistant (GR) cell lines. Initial drug exposure selected for an early resistant phenotype that was independent of drug metabolic pathways. Prolonged drug selection pressure after 16 weeks, led to an induction of cytidine deaminase (CDA) and enhanced drug detoxification. Cross resistance profiles demonstrate approximately 100-fold cross resistance to the pyrimidine nucleoside cytarabine, but no resistance to the same in class agents, azacytidine and decitabine. GR cell lines demonstrated a dose dependent collateral hypersensitivity to class I and II histone deacetylase (HDAC) inhibitors and decreased expression of 3 different global heterochromatin marks, as detected by H4K20me3, H3K9me3 and H3K27me3. Cell morphology of the drug resistant cell lines demonstrated a fibroblastic type appearance with loss of cell-cell junctions and an altered microarray expression pattern, using Gene Ontology (GO) annotation, consistent with progression to an invasive phenotype. Of particular note, the gemcitabine resistant cell lines displayed up to a 15 fold increase in invasive potential that directly correlates with the level of gemcitabine resistance. These findings suggest a mechanistic relationship between chemoresistance and metastatic potential in pancreatic carcinoma and provide evidence for molecular pathways that may be exploited to develop therapeutic strategies for refractory pancreatic cancer.
• Sokoloff, M., Cress, A., Algotar, A. M., Nagle, R., Behnejad, R., Drinkwitz, D., Lodhia, N., Thompson, P., & Stratton, S. P. (2015). DNA damage and repair markers to improve accuracy of prostate cancer diagnosis and identification of aggressive disease.. Journal of Clinical Oncology, 33(15_suppl), e16012-e16012. doi:10.1200/jco.2015.33.15_suppl.e16012
• Algotar, A. M., Cress, A. E., Nagle, R. B., Drinkwitz, D., Lodhia, N. S., Thompson, P. A., & Stratton, S. P. (2014). DNA damage and repair markers (H2AX and RAD51) improve accuracy of prostate cancer diagnosis and improve identification of aggressive disease. CANCER RESEARCH, 74(19).
• Cress, A. E. (2014). Intracellular modifiers of integrin alpha 6p production in aggressive prostate and breast cancer cell lines. Biochemical and Biophysical Research Communications.
• Cress, A. E. (2014). Targeting integrin α6 stimulates curative-type bone metastasis lesions in a xenograft model.. Molecular cancer therapeutics.
• Cress, A. E., Chen, W., Epstein, Y., Mascarenhas, J. B., Wang, T., Zhou, T., Kitties, R., Garcia, J. G., & Jacobson, J. R. (2014). Key Role of Integrin Alpha6/Beta4 in Endothelial Cell Barrier Regulation. CIRCULATION, 130.
• Cress, A. E., Nagle, R. B., Landowski, T. H., Gard, J., Pond, E., Pond, G. D., & Geffre, C. P. (2014). Targeting Integrin α6 Stimulates Curative-Type Bone Metastasis Lesions in a Xenograft Model. Molecular Cancer Therapeutics, 13(6), 1558-1566. doi:10.1158/1535-7163.mct-13-0962
• Das, L., Anderson, T. A., Gard, J., Sroka, I. C., Strautman, S. R., Nagle, R. B., Knudsen, B. S., & Cress, A. (2014). A6B1 integrin internalization is increased by loss of A3B1 expression and promotes migration of human prostate cancer cells. CANCER RESEARCH, 74(19).
• Fu, P., Cress, A. E., Wang, T., Garcia, J. G., & Natarajan, V. (2014). C-Abl Mediated Tyrosine Phosphorylation of Paxillin is Essential for LPS-Induced Endothelial Barrier Dysfunction and Acute Lung Injury. CIRCULATION, 130.
• Kacsinta, A. D., Rubenstein, C. S., Sroka, I. C., Pawar, S., Gard, J. M., Nagle, R. B., & Cress, A. E. (2014). Intracellular modifiers of integrin alpha 6p production in aggressive prostate and breast cancer cell lines. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 454(2), 335-340.
• Kacsinta, A. D., Rubenstein, C. S., Sroka, I. C., Pawar, S., Gard, J. M., Nagle, R. B., & Cress, A. E. (2014). Intracellular modifiers of integrin alpha 6p production in aggressive prostate and breast cancer cell lines. Biochemical and biophysical research communications, 454(2), 335-40.
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  Cancer metastasis is a multi-step process in which tumor cells gain the ability to invade beyond the primary tumor and colonize distant sites. The mechanisms regulating the metastatic process confer changes to cell adhesion receptors including the integrin family of receptors. Our group previously discovered that the α6 integrin (ITGA6/CD49f) is post translationally modified by urokinase plasminogen activator (uPA) and its receptor, urokinase plasminogen activator receptor (uPAR), to form the variant ITGA6p. This variant of ITGA6 is a cleaved form of the receptor that lacks the ligand-binding domain. Although it is established that the uPA/uPAR axis drives ITGA6 cleavage, the mechanisms regulating cleavage have not been defined. Intracellular integrin dependent "inside-out" signaling is a major regulator of integrin function and the uPA/uPAR axis. We hypothesized that intracellular signaling molecules play a role in formation of ITGA6p to promote cell migration during cancer metastasis. In order to test our hypothesis, DU145 and PC3B1 prostate cancer and MDA-MB-231 breast cancer cell lines were treated with small interfering RNA targeting actin and the intracellular signaling regulators focal adhesion kinase (FAK), integrin linked kinase (ILK), and paxillin. The results demonstrated that inhibition of actin, FAK, and ILK expression resulted in significantly increased uPAR expression and ITGA6p production. Inhibition of actin increased ITGA6p, although inhibition of paxillin did not affect ITGA6p formation. Taken together, these results suggest that FAK and ILK dependent "inside-out" signaling, and actin dynamics regulate extracellular production of ITGA6p and the aggressive phenotype.
• Landowski, T. H., Gard, J., Pond, E., Pond, G. D., Nagle, R. B., Geffre, C. P., & Cress, A. E. (2014). Targeting Integrin alpha 6 Stimulates Curative-Type Bone Metastasis Lesions in a Xenograft Model. MOLECULAR CANCER THERAPEUTICS, 13(6), 1558-1566.
• Landowski, T. H., Gard, J., Pond, E., Pond, G. D., Nagle, R. B., Geffre, C. P., & Cress, A. E. (2014). Targeting integrin α6 stimulates curative-type bone metastasis lesions in a xenograft model. Molecular cancer therapeutics, 13(6), 1558-66.
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  Laminin-binding integrin receptors are key mediators of epithelial cell migration and tumor metastasis. Recent studies have demonstrated a role for the α6 integrin (ITGA6/CD49f) in maintaining stem cell compartments within normal bone marrow and in residency of tumors metastatic to bone. In this study, we tested a function-blocking antibody specific for ITGA6, called J8H, to determine if preexisting cancer lesions in bone could be slowed and/or animal survival improved. Human prostate tumors were established by intracardiac injection into male SCID mice and treatment with J8H antibody was initiated after 1 week. Tumor progression was monitored by micro-computed tomography (CT) imaging of skeletal lesions. Animals that received weekly injections of the anti-ITGA6 antibody showed radiographic progression in only 40% of osseous tumors (femur or tibia), compared with control animals, where 80% of the lesions (femur or tibia) showed progression at 5 weeks. Kaplan-Meier survival analysis demonstrated a significant survival advantage for J8H-treated animals. Unexpectedly, CT image analysis revealed an increased proportion of bone lesions displaying a sclerotic rim of new bone formation, encapsulating the arrested lytic lesions in animals that received the anti-ITGA6 antibody treatment. Histopathology of the sclerotic lesions demonstrated well-circumscribed tumor within bone, surrounded by fibrosis. These data suggest that systemic targeting of the ITGA6-dependent function of established tumors in bone may offer a noncytotoxic approach to arrest the osteolytic progression of metastatic prostate cancer, thereby providing a new therapeutic strategy for advanced disease.
• Landowski, T. H., Landowski, T. H., Rubenstein, C. S., Rubenstein, C. S., Nagle, R. B., Nagle, R. B., Landowski, T. H., Landowski, T. H., Gard, J. M., Gard, J. M., Cress, A. E., & Cress, A. E. (2014). Abstract 3152: Integrin α6β1 dependent collective cell migration in prostate cancer metastasis. Cancer Research, 74, 3152-3152. doi:10.1158/1538-7445.am2014-3152
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  Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Investigating the metastasis tactics of cancer cells show some cancers migrate using epithelial to mesenchymal transition (EMT) while others, such as prostate cancer, use collective migration similar to embryonic tubulogenesis. In human tissue specimens, collective migration originates from pre-malignant prostate intraepithelial neoplasia (PIN), occurs in invasive adenocarcinoma and persists in prostate cancer metastatic bone lesions. Successful collective migration requires cell attachment to the surrounding extracellular matrix (ECM), via adhesion receptors, specifically integrins. In prostate cancer, the laminin binding integrins α6β1 and α3β1 are expressed throughout disease progression including metastases. The laminin receptor α6β1 has been shown to be a determinant of prostate tumor cell aggressiveness. Integrin α6β1 undergoes a post-translational modification mediated by the serine protease, urokinase plasminogen activator (uPA) and its receptor uPAR, resulting in a variant form called α6pβ1. In this study, we optimized the growth and retrieval of tumor cells from 3-dimensional matrigel cultures, to determine the role of integrin α6β1, α6pβ1 and α3β1 in DU145 prostate tumor cell collective migration. The DU145 prostate tumor cells embedded in growth factor reduced matrigel, grew robust networks within 24 hours and contained extensive branching structures. The network formation was shown to be dependent on integrin α6β1 expression as determined by specific knockdown of α6β1 over a 24 hour period using 25nM siRNA targeting. Interestingly, the knockdown of α3β1 integrin had no observable effect on the networks. The networks produced by the DU145 cells contained pericellular proteolysis activity along the branches of collective cells as well as at the leading tips of the branches. Laminin 332, its receptor integrin α6β1, and uPAR co-localized on the membranes between cells traveling collectively. Induced production of integrin α6pβ1 increased branching network formation, while blockage of integrin α6pβ1 production impaired collective migration via reduced network formation. Taken together, these observations suggest that collective cell migration through tubulogenesis requires cell adhesion to laminin via integrin α6β1 and production of the α6pβ1 structural variant facilitates this process. Ongoing work is using the model to identify candidate molecular effectors of tubulogenesis as well as pharmacological approaches to block the process and prevent tumor progression. Supported in part by NIH grants CA 23074 and CA 159406. Citation Format: Cynthia S. Rubenstein, Jaime Gard, Raymond B. Nagle, Terry H. Landowski, Anne E. Cress. Integrin α6β1 dependent collective cell migration in prostate cancer metastasis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3152. doi:10.1158/1538-7445.AM2014-3152
• Rubenstein, C. S., Gard, J., Nagle, R. B., Landowski, T. H., & Cress, A. E. (2014). Integrin alpha 6 beta 1 dependent collective cell migration in prostate cancer metastasis. CANCER RESEARCH, 74(19).
• Cress, A., Pawar, S. C., Dougherty, S., Pennington, M. E., Demetriou, M. C., Stea, B. D., Dorr, R. T., & Cress, A. E. (0). alpha6 integrin cleavage: sensitizing human prostate cancer to ionizing radiation. International journal of radiation biology, 83(11-12).
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  The goal was to determine if prostate tumor cells containing a mutant alpha6 integrin would be defective in tumor re-population following clinically relevant fractionated ionizing radiation (IR) treatments.
• Anderson, T. A., Gard, J., Sroka, I. C., Strautman, S. R., Morrissey, C., Knudsen, B. S., & Cress, A. E. (2012). Laminin receptor cross-talk stimulates receptor internalization and migration of human prostate cancer cells. CANCER RESEARCH, 72.
• Chopra, H., Riquer, Z., Cress, A. E., & Sroka, I. C. (2012). Schwann cells promote tumor cell invasion through regulation of the laminin receptorA6B1. CANCER RESEARCH, 72.
• Pond, E. D., Sroka, T. C., Gard, J., Pawar, S. C., Nagle, R. B., & Cress, A. E. (2012). Inadequate repair of ionizing radiation damage: A consequence of the invasive tumor phenotype. CANCER RESEARCH, 72.
• Cress, A. E. (2011). Acquisition of resistance toward HYD1 correlates with a reduction in cleaved α4 integrin expression and a compromised CAM-DR phenotype.. Molecular cancer therapeutics.
• Cress, A. E. (2011). Inhibition of p38-MAPK signaling pathway attenuates breast cancer induced bone pain and disease progression in a murine model of cancer-induced bone pain.. Molecular pain.
• Cress, A. E. (2011). Macrophage-dependent cleavage of the laminin receptor α6β1 in prostate cancer.. Molecular cancer research : MCR.
• Cress, A. E., & Nagle, R. B. (2011). Metastasis Update: Human Prostate Carcinoma Invasion via Tubulogenesis. Prostate Cancer, 2011, 1-10. doi:10.1155/2011/249290
• Cress, A., Nagle, R. B., & Cress, A. E. (2011). Metastasis Update: Human Prostate Carcinoma Invasion via Tubulogenesis. Prostate cancer, 2011.
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  This paper proposes that human prostate carcinoma primarily invades as a cohesive cell collective through a mechanism similar to embryonic tubulogenesis, instead of the popular epithelial-mesenchymal transformation (EMT) model. Evidence supporting a tubulogenesis model is presented, along with suggestions for additional research. Additionally, observations documenting cell adhesion molecule changes in tissue and stromal components are reviewed, allowing for comparisons between the current branching morphogenesis models and the tubulogenesis model. Finally, the implications of this model on prevailing views of therapeutic and diagnostic strategies for aggressive prostatic disease are considered.
• Cress, A., Sroka, I. C., Sandoval, C. P., Chopra, H., Gard, J. M., Pawar, S. C., & Cress, A. E. (2011). Macrophage-dependent cleavage of the laminin receptor α6β1 in prostate cancer. Molecular cancer research : MCR, 9(10).
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  The laminin-binding integrin α6β1 plays a major role in determining the aggressive phenotype of tumor cells during metastasis. Our previous work has shown that cleavage of the α6β1 integrin to produce the structural variant α6pβ1 on tumor cell surfaces is mediated by the serine protease urokinase plasminogen activator (uPA). Cleavage of α6β1 increases tumor cell motility, invasion, and prostate cancer metastasis, and blockage of uPA inhibits α6pβ1 production. In human tumors, uPA and uPAR are expressed in tumor cells and tumor-associated macrophages (TAM). TAMs localize to solid tumors and contribute to increased tumor growth and the metastatic phenotype. In this study, we utilized a coculture system of PC-3 prostate tumor cells and macrophages [12-O-tetradecanoylphorbol-13-acetate (TPA)-differentiated human leukemia HL-60 cells] to investigate the hypothesis that macrophages stimulate the production of the prometastatic variant α6pβ1 on human prostate cancer cells via the uPA/uPAR axis. Our results indicate that adherent macrophages cocultured with PC-3 cells increased PC-3 uPAR mRNA, uPAR cell surface protein expression and α6 integrin cleavage. The stimulation does not require macrophage/tumor cell contact because macrophage conditioned medium is sufficient for increased uPAR transcription and α6 cleavage-dependent PC-3 cell invasion. The increased cleavage was dependent on uPAR because production was blocked by silencing RNA-targeting uPAR. These results indicate that macrophages can stimulate uPA/uPAR production in tumor cells which results in α6 integrin cleavage. These data suggest that TAMs promote prometastatic integrin-dependent pericellular proteolysis.
• Emmons, M. F., Gebhard, A. W., Nair, R. R., Baz, R., McLaughlin, M. L., Cress, A. E., & Hazlehurst, L. A. (2011). Acquisition of Resistance toward HYD1 Correlates with a Reduction in Cleaved alpha 4 Integrin Expression and a Compromised CAM-DR Phenotype. MOLECULAR CANCER THERAPEUTICS, 10(12), 2257-2266.
• Emmons, M. F., Gebhard, A. W., Nair, R. R., Baz, R., McLaughlin, M. L., Cress, A. E., & Hazlehurst, L. A. (2011). Acquisition of resistance toward HYD1 correlates with a reduction in cleaved α4 integrin expression and a compromised CAM-DR phenotype. Molecular cancer therapeutics, 10(12), 2257-66.
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  We recently reported that the β1 integrin antagonist, referred to as HYD1, induces necrotic cell death in myeloma cell lines as a single agent using in vitro and in vivo models. In this article, we sought to delineate the determinants of sensitivity and resistance toward HYD1-induced cell death. To this end, we developed an HYD1 isogenic resistant myeloma cell line by chronically exposing H929 myeloma cells to increasing concentrations of HYD1. Our data indicate that the acquisition of resistance toward HYD1 correlates with reduced levels of the cleaved α4 integrin subunit. Consistent with reduced VLA-4 (α4β1) expression, the resistant variant showed ablated functional binding to fibronectin, VCAM-1, and the bone marrow stroma cell line HS-5. The reduction in binding of the resistant cell line to HS-5 cells translated to a compromised cell adhesion-mediated drug resistant phenotype as shown by increased sensitivity to melphalan- and bortezomib-induced cell death in the bone marrow stroma coculture model of drug resistance. Importantly, we show that HYD1 is more potent in relapsed myeloma specimens than newly diagnosed patients, a finding that correlated with α4 integrin expression. Collectively, these data indicate that this novel d-amino acid peptide may represent a good candidate for pursuing clinical trials in relapsed myeloma and in particular patients with high levels of α4 integrin. Moreover, our data provide further rationale for continued preclinical development of HYD1 and analogues of HYD1 for the treatment of multiple myeloma and potentially other tumors that home and/or metastasize to the bone.
• Emmons, M. F., Gebhard, A., Mclaughlin, M., Cress, A., & Hazlehurst, L. (2011). alpha 4 integrin expression is a determinant for activity of the integrin antagonist HYD1 in multiple myeloma cell lines and patient specimens. CANCER RESEARCH, 71.
• Gebhard, A. W., Emmons, M. F., Argilagos, R. F., McLaughlin, M., Koomen, J., Cress, A. E., & Hazlehurst, L. A. (2011). The forced interaction of alpha 4 integrin and CD44 by the novel peptide HYD1 may be a determinant of HYD1-induced cell death in multiple myeloma. CANCER RESEARCH, 71.
• Pawar, S. C., Sroka, I. C., Pond, E., Pawar, S. C., Nagle, R. B., Gard, J. M., & Cress, A. E. (2011). Abstract 1534: Laminin binding integrin functions in human bone metastatic cancer. Cancer Research, 71, 1534-1534. doi:10.1158/1538-7445.am2011-1534
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  Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL Laminin binding integrins are major determinants of cancer cell migration, invasion and metastasis. In human prostate cancer, both A6B1 and A3B1 laminin-binding integrins are expressed on tumor cell surfaces as observed in either prostatectomy or needle biopsy specimens. In human breast cancer, the A6 integrin is expressed in both primary tumors and in metastatic lesions. Integrin A6B1 is post-translationally modified on the tumor cell surface by urokinase-type plasminogen activator (uPA), producing a structural variant, A6pB1. While the A6pB1 form is devoid of the ligand binding domain, A3B1 is fully functional and not subject to cleavage. In this study, we observed the continued expression of the A6B1 integrin on tumor cell surfaces within human bone metastatic lesions as well as in human tumor xenografts growing within bone. The cleavage function of the A6 integrin was found to be constitutively expressed on nine different cancer cell lines derived from breast, prostate, lung, pancreatic tumors. Interestingly, five different normal cell types expressing the adhesion active A6B1 integrin did not have the constitutive cleavage function. However, the cleavage function in normal cells was inducible by the exogenous addition of uPA. Selection of successive bone resident cancer cells selects for constitutive cleavage of A6 integrin and increasing expression of A3B1 integrin. Taken together, these data suggest that the cleavage function is constitutively activated in cancer specimens and opens the possibility cleavage mediated release from laminin binding may be coordinated with A3 dependent adhesion. (Supported in part by CA23752 and the TACMASS core service of the Arizona Cancer Center). Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1534. doi:10.1158/1538-7445.AM2011-1534
• Sroka, I. C., Gard, J., Pond, E., Pawar, S. C., Nagle, R. B., & Cress, A. E. (2011). Laminin binding integrin functions in human bone metastatic cancer. CANCER RESEARCH, 71.
• Sroka, I. C., Sandoval, C. P., Chopra, H., Gard, J., Pawar, S. C., & Cress, A. E. (2011). Macrophage-Dependent Cleavage of the Laminin Receptor alpha 6 beta 1 in Prostate Cancer. MOLECULAR CANCER RESEARCH, 9(10), 1319-1328.
• Sroka, I. C., Sroka, I. C., Nagle, R. B., Nagle, R. B., Mcdaniel, K., Mcdaniel, K., Gard, J. M., Gard, J. M., Cress, A. E., & Cress, A. E. (2011). Abstract 1543: Potential role of Schwann cells and laminin adhesion in prostate cancer perineural invasion. Cancer Research, 71, 1543-1543. doi:10.1158/1538-7445.am2011-1543
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  Prostate tumor cell migration and invasion of prostatic nerves, a process known as perineural invasion (PNI), is a major route for extracapsular extension during prostate cancer metastasis. The clinical significance of perineural invasion as a predictor for poor patient outcome has been extensively documented. The laminin family of extracellular matrix glycoproteins lines the neural route and plays a prominent role in the development and maintenance of the peripheral nervous system. Evidence has shown that the laminin receptor integrin α6β1 promotes prostate tumor cell migration and invasion in aggressive disease. We hypothesize that α6β1 regulates prostate cancer perineural invasion by regulating laminin dependent adhesion and migration on nerves. In this study we demonstrate remarkable integrin α6 expression in prostate tumors undergoing active perineural invasion using immunohistochemical analysis of human prostate tumors. The expression of integrin α6 correlates with expression of the laminin subunits α5 and γ1 which comprise laminin-511, the major ligand for integrin α6β1. The laminin extracellular matrix on the peripheral nerves of the prostate is derived from Schwann cells during the myelination process. We demonstrate here that Schwann cells are highly expressed in prostatic nerves using immunohistochemical analysis of the Schwann cell specific marker S-100, suggesting that tumor cells invade myelinated nerves. We also demonstrate two specific patterns of prostate tumor nerve invasion along prostatic nerves. Invasion along the external surface of the nerve sheath or in the perineural space maintains nerve structure while invasion of the endoneural space disrupts nerve structure and displaces Schwann cells. This suggests that tumor interactions with nerves and Schwann cells may illicit nerve damage during the PNI process. Using a DU-145 prostate tumor and neuron/glial cell in vitro co-culture model we demonstrate that DU-145 cells (cytokeratin positive) migrate towards glial cells (GFAP positive) and neurons (Beta-III tubulin positive) isolated from dorsal root ganglia of male SCID mice. Mature cultures of the prostate tumor cells and neuron/glial cells demonstrate distinct islands of growth. The patterns of colony formation were non-random and interspersed colonies of tumor and nerve cells were not observed. Taken together, our immunohistochemical results suggest that perineural invasion of laminin coated nerves is regulated by the laminin adhesion receptor α6β1. Our in vitro model system demonstrates that tumor cells migrate to neuron/glial cells and dominate the co-culture, correlating with our immunohistochemical data. We expect that blocking α6β1 function in this system will impede prostate tumor cell migration and invasion in the nerve environment. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1543. doi:10.1158/1538-7445.AM2011-1543
• Sukhtankar, D., Okun, A., Chandramouli, A., Nelson, M. A., Vanderah, T. W., Cress, A. E., Porreca, F., & King, T. (2011). Inhibition of p38-MAPK signaling pathway attenuates breast cancer induced bone pain and disease progression in a murine model of cancer-induced bone pain. MOLECULAR PAIN, 7.
• Sukhtankar, D., Okun, A., Chandramouli, A., Nelson, M. A., Vanderah, T. W., Cress, A. E., Porreca, F., & King, T. (2011). Inhibition of p38-MAPK signaling pathway attenuates breast cancer induced bone pain and disease progression in a murine model of cancer-induced bone pain. Molecular pain, 7, 81.
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  Mechanisms driving cancer-induced bone pain are poorly understood. A central factor implicated to be a key player in the process of tumorigenesis, osteoclastogenesis and nociception is p38 MAPK. We determined the role of p38 MAPK in a mouse model of breast cancer induced bone pain in which mixed osteolytic and osteoblastic remodeling occurs.
• Cress, A. E. (2010). Role for DNA methylation in the regulation of miR-200c and miR-141 expression in normal and cancer cells.. PloS one.
• Cress, A. E. (2010). The laminin binding integrin alpha6beta1 in prostate cancer perineural invasion.. Journal of cellular physiology.
• Cress, A., Sroka, I. C., Anderson, T. A., McDaniel, K. M., Nagle, R. B., Gretzer, M. B., & Cress, A. E. (2010). The laminin binding integrin alpha6beta1 in prostate cancer perineural invasion. Journal of cellular physiology, 224(2).
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  Metastasizing prostate tumor cells invade along nerves innervating the encapsulated human prostate gland in a process known as perineural invasion. The extracellular matrix laminin class of proteins line the neural route and tumor cells escaping from the gland express the laminin binding integrin alpha6beta1 as a prominent cell surface receptor. Integrin alpha6beta1 promotes aggressive disease and supports prostate tumor cell metastasis to bone. Laminins and their integrin receptors are necessary for the development and maintenance of the peripheral nervous system, indicating the potential role for integrin receptors in directing prostate tumor cell invasion on nerves during perineural invasion.
• Nair, R. R., Mclaughlin, M. L., Hazlehurst, L. A., Eschrich, S. A., Emmons, M. F., & Cress, A. E. (2010). Abstract 1547: Acquisition of resistance to the β1 integrin antagonist HYD1 correlates with decreased α4, α6, αV and β1 integrin expression and decreased adhesion to fibronectin, VCAM-1 and vitronectin. Cancer Research, 70, 1547-1547. doi:10.1158/1538-7445.am10-1547
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  Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Our laboratory recently reported that the D amino acid-containing β1 integrin inhibitory peptide HYD1 causes caspase independent cell death in multiple myeloma (MM) cell lines as a single agent in vitro and in vivo (Nair et al Molecular Cancer Therapeutics, 2009). Exposure of MM cells to HYD1 induces autophagy, but autophagy was shown to contribute to survival following HYD1 treatment. HYD1 treatment induces cell death in MM cell lines indicative of necrosis. These characteristics included: (a) an increase in the levels of reactive oxygen species (ROS), (b) ATP depletion and (c) loss of mitochondrial membrane potential. To further characterize the mechanism of HYD1 induced cell death an isogenic drug resistant variant was developed. This variant, referred to as H929-60, was developed by chronically exposing parental H929 cells to increasing doses of HYD1. Acquired resistance correlated with decreased HYD1 induced TO-PRO-3 positivity, decreased levels of ROS, reduced ATP depletion and reduced mmp loss compared to the H929 cell line. Additionally, the drug resistant variant H929-60 cells displayed decreased cell surface binding of FAM-HYD1 compared to H929 cells. These data indicate that decreased expression of the binding target(s) of HYD1 may contribute to drug resistance. Gene expression profiling (GEP) was used as an unbiased approach to identify changes in gene expression that may contribute to HYD1 resistance. Differentially expressed genes were mined in silico using the pathway analysis programs Ingenuity and GeneGo. The integrin signaling hub showed substantial changes in gene expression in the drug resistant variant compared to the parental cell line. Specifically, α4, α6, αV and β1 integrins were all down regulated along with the integrin associated protein CD47 in the HYD1 resistant cell line. These changes were consistent with decreased protein expression of integrins on the cell membrane and decreased functional adhesion to fibronectin, vitronectin and VCAM-1. Reducing the expression of α4 integrin using shRNA in the parental H929 cell line conferred partial resistance to HYD1 induced cell death. Together our data show that the acquisition of resistance to HYD1 correlated with decreased integrin expression and functional binding to extracellular matrixes. Future studies aim to determine whether a decrease in α6, αV and β1 integrins on the cell surface contributes to HYD1 resistance. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1547.
• Pawar, S. C., Abril, E. R., Sroka, I. C., Sandoval, C. P., Pawar, S. C., Nagle, R. B., Gard, J. M., Cress, A. E., & Abril, E. R. (2010). Abstract 534: Invasive human prostate cancer and integrin cleavage: Potential role of uPAR contributed by macrophages. Cancer Research, 70, 534-534. doi:10.1158/1538-7445.am10-534
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  Laminin binding integrins are major determinants of cancer cell migration, invasion and metastasis. Our previous work showed that integrin α6β1 is cleaved on the tumor cell surface by urokinase-type plasminogen activator (uPA), producing a structural variant, α6pβ1. Urokinase plasminogen activator receptor (uPAR) is a regulatory determinant for α6 integrin cleavage since uPAR interacts with integrin and activates uPA. Previous work by others has shown that both human tumor tissue and tumor infiltrating macrophages express uPAR . In the present study, we investigated the potential of uPAR to co-localize with α6β1 integrin in human prostate cancer cells (DU-145) and whether uPAR expressed on the macrophage cell surface mediates integrin α6β1 cleavage. The results indicated uPAR and integrin α6β1 were co-expressed in invasive human prostate carcinoma. Co-localization of uPAR and integrin α6β1 occurred in some tumor cells in vitro, consistent with reports indicating a variable distribution in human tumors. The culture of macrophages with tumor cells resulted in an increased cleavage of integrin α6 as detected by immuno precipitation followed by western blot analysis. Taken together, these data suggest that integrin α6 cleavage is directed by the receptor for uPA, uPAR, contributed either by the tumor cell itself or tumor cell contact with macrophages. (Supported in part by CA 56666, CA23752, T34 GM008718 and the TACMASS core service of the Arizona Cancer Center) Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 534.
• Pawar, S. C., Nagle, R. B., Sroka, T. C., Pond, G. D., Pawar, S. C., Nagle, R. B., & Cress, A. E. (2010). Blocking Integrin Function Combined with Ionizing Radiation for Eradication of Bone Metastasis. International Journal of Radiation Oncology Biology Physics, 78(3), S622-S623. doi:10.1016/j.ijrobp.2010.07.1448
• Sroka, I. C., Anderson, T. A., McDaniel, K. M., Nagle, R. B., Gretzer, M. B., & Cress, A. E. (2010). The Laminin Binding Integrin alpha 6 beta 1 in Prostate Cancer Perineural Invasion. JOURNAL OF CELLULAR PHYSIOLOGY, 224(2), 283-288.
• Vrba, L., Jensen, T. J., Garbe, J. C., Heimark, R. L., Cress, A. E., Dickinson, S., Stampfer, M. R., & Futscher, B. W. (2010). Role for DNA Methylation in the Regulation of miR-200c and miR-141 Expression in Normal and Cancer Cells. PLOS ONE, 5(1).
• Vrba, L., Jensen, T. J., Garbe, J. C., Heimark, R. L., Cress, A. E., Dickinson, S., Stampfer, M. R., & Futscher, B. W. (2010). Role for DNA methylation in the regulation of miR-200c and miR-141 expression in normal and cancer cells. PloS one, 5(1), e8697.
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  The microRNA-200 family participates in the maintenance of an epithelial phenotype and loss of its expression can result in epithelial to mesenchymal transition (EMT). Furthermore, the loss of expression of miR-200 family members is linked to an aggressive cancer phenotype. Regulation of the miR-200 family expression in normal and cancer cells is not fully understood.
• Cress, A. E. (2009). Extracellular engagement of alpha6 integrin inhibited urokinase-type plasminogen activator-mediated cleavage and delayed human prostate bone metastasis.. Cancer research.
• Cress, A. E. (2009). HYD1-induced increase in reactive oxygen species leads to autophagy and necrotic cell death in multiple myeloma cells.. Molecular cancer therapeutics.
• Cress, A. E. (2009). Human Cell Surface Receptors as Molecular Imaging Candidates for Metastatic Prostate Cancer.. The open prostate cancer journal.
• Cress, A. E. (2009). Transient dephosphorylation of p53 serine 376 as an early response to ionizing radiation.. Radiation research.
• Cress, A., Ports, M. O., Nagle, R. B., Pond, G. D., & Cress, A. E. (2009). Extracellular engagement of alpha6 integrin inhibited urokinase-type plasminogen activator-mediated cleavage and delayed human prostate bone metastasis. Cancer research, 69(12).
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  Expression of alpha(6) integrin, a laminin receptor, on tumor cell surfaces is associated with reduced patient survival and increased metastasis in a variety of tumors. In prostate cancer, tumor extracapsular escape occurs in part via laminin-coated nerves and vascular dissemination, resulting in clinically significant bone metastases. We previously identified a novel form of alpha(6) integrin, called alpha(6)p, generated by urokinase-type plasminogen activator-dependent cleavage of the laminin-binding domain from the tumor cell surface. Cleavage increased laminin-dependent migration. Currently, we used the known conformation sensitivity of integrin function to determine if engagement of the extracellular domain inhibited integrin cleavage and the extravasation step of metastasis. We show that alpha(6) integrin was present on prostate carcinoma escaping the gland via nerves. Both endogenous and inducible levels of alpha(6)p were inhibited by engaging the extracellular domain of alpha(6) with monoclonal antibody J8H. J8H inhibited tumor cell invasion through Matrigel. A severe combined immunodeficient mouse model of extravasation and bone metastasis produced detectable, progressive osteolytic lesions within 3 weeks of intracardiac injections. Injection of tumor cells, pretreated with J8H, delayed the appearance of metastases. Validation of the alpha(6) cleavage effect on extravasation was confirmed through a genetic approach using tumor cells transfected with uncleavable alpha(6) integrin. Uncleavable alpha(6) integrin significantly delayed the onset and progression of osseous metastases out to six weeks post-injection. The results suggest that alpha(6) integrin cleavage permits extravasation of human prostate cancer cells from circulation to bone and can be manipulated to prevent metastasis.
• Cress, A., Sroka, I. C., Pond, G. D., Nagle, R. B., Porreca, F., King, T., Pestano, G., Futscher, B. W., Gard, J. M., Riley, J., & Cress, A. E. (2009). Human Cell Surface Receptors as Molecular Imaging Candidates for Metastatic Prostate Cancer. The open prostate cancer journal, 2.
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  Existing clinical imaging procedures lack sensitivity and specificity in detecting early prostate cancer bone metastatic lesions. In this study, we developed a highly reproducible bone metastasis xenograft model and identified possible molecular imaging candidates for detecting early bone metastatic lesions. Bone trophic human prostate cells (PC-3B1) were isolated and characterized for their ability to reach bone after intracardiac injection into SCID mice. The appearances of skeletal metastases were evaluated using digital radiographic imaging and confirmed by necropsy and histology. The PC-3B1 cells retain a bone homing phenotype after long term propagation in tissue culture and exhibit progressive bone lesions within 3 weeks following intracardiac injection. Comparative transcription signatures of PC-3 and PC-3B1 cells were determined using a cancer specific microarray and confirmed by RT-PCR analysis. The analysis identified increased expression of four cell surface molecules in PC-3B1 cells that may be suitable as molecular imaging candidates to detect bone micro metastases.
• Nair, R. R., Emmons, M. F., Cress, A. E., Argilagos, R. F., Lam, K., Kerr, W. T., Wang, H. G., Dalton, W. S., & Hazlehurst, L. A. (2009). HYD1-induced increase in reactive oxygen species leads to autophagy and necrotic cell death in multiple myeloma cells. Molecular cancer therapeutics, 8(8), 2441-51.
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  HYD1 is a D-amino acid peptide that was previously shown to inhibit adhesion of prostate cancer cells to the extracellular matrix. In this study, we show that in addition to inhibiting adhesion of multiple myeloma (MM) cells to fibronectin, HYD1 induces cell death in MM cells as a single agent. HYD1-induced cell death was necrotic in nature as shown by: (a) decrease in mitochondrial membrane potential (Deltapsi(m)), (b) loss of total cellular ATP, and (c) increase in reactive oxygen species (ROS) production. Moreover, HYD1 treatment does not result in apoptotic cell death because it did not trigger the activation of caspases or the release of apoptosis-inducing factor and endonuclease G from the mitochondria, nor did it induce double-stranded DNA breaks. HYD1 did initiate autophagy in cells; however, autophagy was found to be an adaptive response contributing to cell survival rather than the cause of cell death. We were further able to show that N-acetyl-L-cysteine, a thiol-containing free radical scavenger, partially protects MM cells from HYD1-induced death. Additionally, N-acetyl-L-cysteine blocked HYD1-induced as well as basal levels of autophagy, suggesting that ROS can potentially trigger both cell death and cell survival pathways. Taken together, our data describe an important role of ROS in HYD1-induced necrotic cell death in MM cells.
• Nair, R. R., Emmons, M. F., Cress, A. E., Argilagos, R. F., Lam, K., Kerr, W. T., Wang, H., Dalton, W. S., & Hazlehurst, L. A. (2009). HYD1-induced increase in reactive oxygen species leads to autophagy and necrotic cell death in multiple myeloma cells. MOLECULAR CANCER THERAPEUTICS, 8(8), 2441-2451.
• Ports, M. O., Nagle, R. B., Pond, G. D., & Cress, A. E. (2009). Extracellular Engagement of alpha(6) Integrin Inhibited Urokinase-Type Plasminogen Activator-Mediated Cleavage and Delayed Human Prostate Bone Metastasis. CANCER RESEARCH, 69(12), 5007-5014.
• Ports, M. O., Nagle, R. B., Pond, G. D., & Cress, A. E. (2009). Extracellular engagement of alpha 6 integrin blocks uPA mediated cleavage and prevents human prostate bone metastasis. CLINICAL & EXPERIMENTAL METASTASIS, 26(7), 871-871.
• Warters, R. L., Gaffney, D. K., Kramer, G. F., Martinez, J. D., & Cress, A. E. (2009). Transient Dephosphorylation of p53 Serine 376 as an Early Response to Ionizing Radiation. RADIATION RESEARCH, 171(6), 725-734.
• Warters, R. L., Gaffney, D. K., Kramer, G. F., Martinez, J. D., & Cress, A. E. (2009). Transient dephosphorylation of p53 serine 376 as an early response to ionizing radiation. Radiation research, 171(6), 725-34.
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  In a previous paper we reported that the cytoplasmic sequestered p53 in cells of the SK-N-SH neuroblastoma cell line could be induced to translocate to the nucleus by exposure to ionizing radiation. We have extended these studies to determine the fate of p53 in HCT116 colorectal carcinoma cells where constitutive p53 protein resides in the nucleus. A continuous increase in the nuclear p53 protein was observed in irradiated cells beginning 1 h after irradiation that persisted for 8 h. Surprisingly, immunofluorescence microscopy revealed a transient, rapid and sensitive increase in a radiation-induced nuclear dephosphorylated p53 using antibody PAb421, which detects p53 when serine 376 is dephosphorylated. The PAb421 epitope was detectable after exposure to radiation doses as low as 0.5 cGy and was 10 to 20 times more sensitive compared to detection of p53 protein levels. The results are consistent with a radiation-induced, sensitive and rapid dephosphorylation of p53 at serine 376. The rapid increase in the nuclear PAb421 epitope was blocked by the protein serine phosphatase inhibitor calyculin A but was not blocked by the protein synthesis inhibitor cycloheximide, suggesting that serine 376 was dephosphorylated by protein serine phosphatase 1 or 2A acting on pre-existing p53 protein. The data suggest that dephosphorylation of serine 376 on constitutive nuclear p53 is a sensitive and early signaling event in the response of cells to DNA damage induced by ionizing radiation.
• Cress, A. E. (2008). A comprehensive space management model for facilitating programmatic research.. Academic medicine : journal of the Association of American Medical Colleges.
• Cress, A. E. (2008). Integrin A6 Cleavage in Mouse Skin Tumors.. The open cancer journal.
• Cress, A. E. (2008). Simplified purification procedure of laminin-332 and laminin-511 from human cell lines.. Biochemical and biophysical research communications.
• Cress, A. E. (2008). Supporting the academic mission in an era of constrained resources: approaches at the University of Arizona College of Medicine.. Academic medicine : journal of the Association of American Medical Colleges.
• Cress, A. E. (2008). The role of alpha 6 integrin in prostate cancer migration and bone pain in a novel xenograft model.. PloS one.
• Cress, A., King, T. E., Pawar, S. C., Majuta, L., Sroka, I. C., Wynn, D., Demetriou, M. C., Nagle, R. B., Porreca, F., & Cress, A. E. (2008). The role of alpha 6 integrin in prostate cancer migration and bone pain in a novel xenograft model. PloS one, 3(10).
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  Of the estimated 565,650 people in the U.S. who will die of cancer in 2008, almost all will have metastasis. Breast, prostate, kidney, thyroid and lung cancers metastasize to the bone. Tumor cells reside within the bone using integrin type cell adhesion receptors and elicit incapacitating bone pain and fractures. In particular, metastatic human prostate tumors express and cleave the integrin A6, a receptor for extracellular matrix components of the bone, i.e., laminin 332 and laminin 511. More than 50% of all prostate cancer patients develop severe bone pain during their remaining lifetime. One major goal is to prevent or delay cancer induced bone pain. We used a novel xenograft mouse model to directly determine if bone pain could be prevented by blocking the known cleavage of the A6 integrin adhesion receptor. Human tumor cells expressing either the wildtype or mutated A6 integrin were placed within the living bone matrix and 21 days later, integrin expression was confirmed by RT-PCR, radiographs were collected and behavioral measurements of spontaneous and evoked pain performed. All animals independent of integrin status had indistinguishable tumor burden and developed bone loss 21 days after surgery. A comparison of animals containing the wild type or mutated integrin revealed that tumor cells expressing the mutated integrin resulted in a dramatic decrease in bone loss, unicortical or bicortical fractures and a decrease in the ability of tumor cells to reach the epiphyseal plate of the bone. Further, tumor cells within the bone expressing the integrin mutation prevented cancer induced spontaneous flinching, tactile allodynia, and movement evoked pain. Preventing A6 integrin cleavage on the prostate tumor cell surface decreased the migration of tumor cells within the bone and the onset and degree of bone pain and fractures. These results suggest that strategies for blocking the cleavage of the adhesion receptors on the tumor cell surface can significantly prevent cancer induced bone pain and slow disease progression within the bone. Since integrin cleavage is mediated by Urokinase-type Plasminogen Activator (uPA), further work is warranted to test the efficacy of uPA inhibitors for prevention or delay of cancer induced bone pain.
• Cress, A., Sroka, I. C., Chen, M. L., & Cress, A. E. (2008). Simplified purification procedure of laminin-332 and laminin-511 from human cell lines. Biochemical and biophysical research communications, 375(3).
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  Laminins are glycoproteins expressed in the basement membrane of multiple epithelial tissues. Previously described purification procedures for the human laminin variants laminin-5 (LN-332) and laminin-10 (LN-511) use tissue as starting material and have multiple steps. We demonstrate a two-step laminin immunoaffinity purification method to produce consistent quantities of intact and biologically active LN-332 and LN-511 from human keratinocyte (HaCaT) and human lung carcinoma (A549) cell lines, respectively. The purification of LN-332 and LN-551 was demonstrated by PAGE analysis, silver staining and Western blot analysis. The purification procedure includes instruction on removing a cell adhesion contaminant known as galectin-3 binding protein from purified LN-511. The biological activity of purified laminin was tested in a standard cell adhesion assay and compared to commercially available LN-111. This rapid and reproducible purification method will contribute to understanding the role of LN-332 and LN-511 in cell behavior, signaling, and gene expression.
• Demetriou, M. C., Kwei, K. A., Powell, M. B., Nagle, R. B., Bowden, G. T., & Cress, A. E. (2008). Integrin A6 Cleavage in Mouse Skin Tumors. The open cancer journal, 2, 1-4.
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  We have previously identified a structural variant of the α6 integrin (Laminin receptor) called α6p. The α6p variant is a 70 kDa form of the full-length α6 integrin (140 kDa) that remains paired with either the β1 or β4 subunit on the cell surface. α6p is produced by urokinase-type plasminogen activator (uPA), which removes the extracellular β-barrel domain while the receptor is on the cell surface. The α6p integrin was present in human prostate cancer tissue but not in normal tissue and the cleavage of the α6 integrin extracellular domain promotes tumor cell invasion and migration on laminin. The objective of the present study was to determine whether the α6p integrin is observed in other models of carcinogenesis. Our results indicate detectable low levels of α6p in normal mouse skin, and comparatively elevated levels in mouse papillomas and squamous cell carcinomas induced by DMBA, TPA and MNNG treatments. Furthermore, we have found that α6p was present at high levels in skin melanomas of transgenic mice that over express activated Ha-ras under the control of the tyrosinase promoter. Finally, subcutaneous injection into athymic nude mice of a malignant mouse keratinocyte derived cell line (6M90) that is α6p negative, results in the development of tumors that contain α6p integrin. The latter results indicate that α6p is induced in vivo suggesting that the tumor microenvironment plays a major role in the production of α6p. Taken together, these data suggest that the cell surface cleavage of the α6 integrin may be a novel mechanism of integrin regulation and might be an important step during skin tissue remodeling and during carcinogenesis.
• Demetriou, M. C., Stylianou, P., Andreou, M., Yiannikouri, O., Tsaprailis, G., Cress, A. E., & Skourides, P. (2008). Spatially and temporally regulated alpha 6 integrin cleavage during Xenopus laevis development. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 366(3), 779-785.
• Demetriou, M. C., Stylianou, P., Andreou, M., Yiannikouri, O., Tsaprailis, G., Cress, A. E., & Skourides, P. (2008). Spatially and temporally regulated alpha6 integrin cleavage during Xenopus laevis development. Biochemical and biophysical research communications, 366(3), 779-85.
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  The alpha6 integrin is essential for early nervous system development in Xenopus laevis. We have previously reported a uPA cleaved form of integrin alpha6 (alpha6p), in invasive human prostate cancer tissue, whose presence correlates with increased migration and invasive capacity. We now report that alpha6 is cleaved during the normal development of Xenopus in a spatially and temporally controlled manner. In addition, unlike normal mammalian tissues, which lack alpha6p, the major form of the alpha6 integrin present in adult Xenopus is alpha6p. The protease responsible for the cleavage in mammals, uPA, is not involved in the cleavage of Xenopus alpha6. Finally, overexpression of a mammalian alpha6 mutant which cannot be cleaved leads to developmental abnormalities suggesting a potential role for the cleavage in development.
• Joiner, K. A., Libecap, A., Cress, A. E., Wormsley, S., St Germain, P., Berg, R., & Malan, P. (2008). Supporting the academic mission in an era of constrained resources: approaches at the University of Arizona College of Medicine. Academic medicine : journal of the Association of American Medical Colleges, 83(9), 837-44.
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  The authors describe initiatives at the University of Arizona College of Medicine to markedly expand faculty, build research along programmatic lines, and promote a new, highly integrated medical school curriculum. Accomplishing these goals in this era of declining resources is challenging. The authors describe their approaches and outcomes to date, derived from a solid theoretical framework in the management literature, to (1) support research faculty recruitment, emphasizing return on investment, by using net present value to guide formulation of recruitment packages, (2) stimulate efficiency and growth through incentive plans, by using utility theory to optimize incentive plan design, (3) distribute resources to support programmatic growth, by allocating research space and recruitment dollars to maximize joint hires between units with shared interests, and (4) distribute resources from central administration to encourage medical student teaching, by aligning state dollars to support a new integrated organ-system based-curriculum. Detailed measurement is followed by application of management principles, including mathematical modeling, to make projections based on the data collected. Although each of the initiatives was developed separately, they are linked functionally and financially, and they are predicated on explicitly identifying opportunity costs for all major decisions, to achieve efficiencies while supporting growth. The overall intent is to align institutional goals in education, research, and clinical care with incentives for unit heads and individual faculty to achieve those goals, and to create a clear line of sight between expectations and rewards. Implementation is occurring in a hypothesis-driven fashion, permitting testing and refinement of the strategies.
• Joiner, K. A., Libecap, A., Cress, A. E., Wormsley, S., St, G. P., Berg, R., & Malan, P. (2008). Supporting the Academic Mission in an Era of Constrained Resources: Approaches at the University of Arizona College of Medicine. ACADEMIC MEDICINE, 83(9), 837-844.
• King, T. E., Pawar, S. C., Majuta, L., Sroka, I. C., Wynn, D., Demetriou, M. C., Nagle, R. B., Porreca, F., & Cress, A. E. (2008). The Role of Alpha 6 Integrin in Prostate Cancer Migration and Bone Pain in a Novel Xenograft Model. PLOS ONE, 3(10).
• Libecap, A., Wormsley, S., Cress, A., Matthews, M., Souza, A., & Joiner, K. A. (2008). A Comprehensive Space Management Model for Facilitating Programmatic Research. ACADEMIC MEDICINE, 83(3), 207-216.
• Sroka, I. C., Chen, M. L., & Cress, A. E. (2008). Simplified purification procedure of laminin-332 and laminin-511 from human cell lines. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 375(3), 410-413.
• Cress, A. E. (2007). Integrin α6 cleavage: A novel modification to modulate cell migration. Experimental Cell Research.
• Cress, A. E. (2007). Morphine treatment accelerates sarcoma-induced bone pain, bone loss, and spontaneous fracture in a murine model of bone cancer.. Pain.
• Cress, A. E. (2007). Spatially and temporally regulated alpha6 integrin cleavage during Xenopus laevis development.. Biochemical and biophysical research communications.
• Cress, A. E. (2007). alpha6 integrin cleavage: sensitizing human prostate cancer to ionizing radiation.. International journal of radiation biology.
• Cress, A., Pawar, S. C., Demetriou, M. C., Nagle, R. B., Bowden, G. T., & Cress, A. E. (2007). Integrin alpha6 cleavage: a novel modification to modulate cell migration. Experimental cell research, 313(6).
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  Integrins play a major role in cell adhesion and migration. Previous work reported that a cleaved form of integrin alpha6 (alpha6p) was detected in invasive human prostate cancer tissue, absent in normal prostate tissue and was produced by urokinase-type Plasminogen Activator (uPA) in a plasmin-independent manner. Using site-directed mutagenesis we identified amino acid residues R594 and R595, located in the "stalk" region of integrin alpha6, as essential for cleavage. The cleavage site is located on the extracellular region of the protein between the beta-barrel domain and the thigh domain. Prostate cancer cells (PC3N) were stably transfected to overexpress the cleavable, wild-type (PC3N-alpha6-WT) or the non-cleavable form of integrin alpha6 (PC3N-alpha6-RR). The number of cells invading laminin 111- and laminin 332-coated filters by PC3N-alpha6-WT cells increased by threefold as compared to PC3N-alpha6-RR cells. Plasminogen activator inhibitor-1 (PAI-1) reduced the invasion of PC3N-alpha6-WT cells by approximately 42% through laminin 332-coated filters and plasmin inhibitor aprotinin had no significant effect. Linear cell migration increased production of integrin alpha6p in the PC3N-alpha6-WT cells and not in the PC3N-alpha6-RR cells and 32% of the PC3N-alpha6-WT cells migrated on laminin 111 in the linear migration assay as compared to the 5% PC3N-alpha6-RR cells. These data taken together suggest that the uPA-mediated cell surface cleavage of the alpha6 integrin extracellular domain is involved in tumor cell invasion and migration on laminin.
• King, T., Vardanyan, A., Majuta, L., Melemedjian, O., Nagle, R., Cress, A. E., Vanderah, T. W., Lai, J., & Porreca, F. (2007). Morphine treatment accelerates sarcoma-induced bone pain, bone loss, and spontaneous fracture in a murine model of bone cancer. PAIN, 132(1-2), 154-168.
• King, T., Vardanyan, A., Majuta, L., Melemedjian, O., Nagle, R., Cress, A. E., Vanderah, T. W., Lai, J., & Porreca, F. (2007). Morphine treatment accelerates sarcoma-induced bone pain, bone loss, and spontaneous fracture in a murine model of bone cancer. Pain, 132(1-2), 154-68.
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  Metastatic bone cancer causes severe pain that is primarily treated with opioids. A model of bone cancer pain in which the progression of cancer pain and bone destruction is tightly controlled was used to evaluate the effects of sustained morphine treatment. In cancer-treated mice, morphine enhanced, rather than diminished, spontaneous, and evoked pain; these effects were dose-dependent and naloxone-sensitive. SP and CGRP positive DRG cells did not differ between sarcoma or control mice, but were increased following morphine in both groups. Morphine increased ATF-3 expression only in DRG cells of sarcoma mice. Morphine did not alter tumor growth in vitro or tumor burden in vivo but accelerated sarcoma-induced bone destruction and doubled the incidence of spontaneous fracture in a dose- and naloxone-sensitive manner. Morphine increased osteoclast activity and upregulated IL-1 beta within the femurs of sarcoma-treated mice suggesting enhancement of sarcoma-induced osteolysis. These results indicate that sustained morphine increases pain, osteolysis, bone loss, and spontaneous fracture, as well as markers of neuronal damage in DRG cells and expression of pro-inflammatory cytokines. Morphine treatment may result in "add-on" mechanisms of pain beyond those engaged by sarcoma alone. While it is not known whether the present findings in this model of osteolytic sarcoma will generalize to other cancers or opioids, the data suggest a need for increased understanding of neurobiological consequences of prolonged opioid exposure which may allow improvements in the use of opiates in the effective management of cancer pain.
• Pawar, S. C., Demetriou, M. C., Nagle, R. B., Bowden, G. T., & Cress, A. E. (2007). Integrin alpha 6 cleavage: A novel modification to modulate cell migration. EXPERIMENTAL CELL RESEARCH, 313(6), 1080-1089.
• Pawar, S. C., Dougherty, S., Pennington, M. E., Demetriou, M. C., Stea, B. D., Dorr, R. T., & Cress, A. E. (2007). alpha 6 Integrin Cleavage: Sensitizing human prostate cancer to ionizing radiation. INTERNATIONAL JOURNAL OF RADIATION BIOLOGY, 83(11-12), 761-767.
• Cress, A. E. (2006). Integrin-dependent amplification of the G2 arrest induced by ionizing radiation.. The Prostate.
• Cress, A. E. (2006). Synthetic D-amino acid peptide inhibits tumor cell motility on laminin-5.. Carcinogenesis.
• Cress, A. E. (2006). The minimum element of a synthetic peptide required to block prostate tumor cell migration.. Cancer biology & therapy.
• Cress, A. E. (2006). Tumor progression and metastasis from genetic to microenvironmental determinants: a workshop of the tumor progression and metastasis NIH study section in honor of Dr. Martin L. Padarathsingh, May 31, 2006, Georgetown, Washington, DC.. Cancer biology & therapy.
• Cress, A., Kremer, C. L., Schmelz, M., & Cress, A. E. (2006). Integrin-dependent amplification of the G2 arrest induced by ionizing radiation. The Prostate, 66(1).
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  The progressive loss of laminin 5 and the alpha6beta4 integrin is a characteristic of the transition of prostatic intraepithelial neoplasia (PIN) to invasive human prostate cancer. Our objective was to determine if the loss of the interaction with laminin 5 would influence the ability of human epithelial cells to respond to DNA damage. Three cellular damage responses to ionizing radiation (IR) were analyzed including G2 progression, cdc2 phosphorylation, and cell survival. The adhesion of normal human prostate epithelial cells to laminin 5 amplified the G2 arrest induced by IR, and depends on a known cell binding domain of laminin 5. The alteration of G2 arrest was confirmed by an inhibition of phospho-cdc2 nuclear translocation. In contrast, a prostate epithelial cancer cell line blocked in G2 independent of adhesion to laminin 5. The survival of these cell lines in response to IR was unaffected by adhesion to laminin 5. These results suggest that cell adhesion to laminin 5 in normal cells will amplify the IR induced G2 cell cycle progression block without altering cell survival. The loss of laminin 5 and the alpha6beta4 integrin in PIN lesions may contribute to the selection and progression of genetically unstable cell types via attenuation of a DNA damage induced G2 arrest.
• Cress, A., Sroka, T. C., Marik, J., Pennington, M. E., Lam, K. S., & Cress, A. E. (2006). The minimum element of a synthetic peptide required to block prostate tumor cell migration. Cancer biology & therapy, 5(11).
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  Human prostate tumor cell invasion and metastasis are dependent in part on cell adhesion to extracellular matrix proteins and cell migration. We previously identified a synthetic D-amino acid tumor cell adhesion peptide called HYD1 (kikmviswkg) that supported adhesion of tumor cells derived from breast, prostate, ovary and pancreas tissue. Alanine substitution analysis and a peptide deletion strategy were used to determine the minimal element of HYD1 necessary for bioactivity in a prostate cancer cell line called PC3N. Bioactivity was measured by assays of cell adhesion, migration and ERK signaling. The most potent element of HYD1 necessary to support cell adhesion was kmvixw, the block to migration required xkmviswxx and activation of ERK signaling required ikmviswxx. The shortest sequence active in all three assays was iswkg. The HYD1 peptide contains overlapping elements required for adhesion, blocking migration and the activation of ERK signaling. These linear peptide sequences provide the starting point for development of novel compounds to target cancer cell adhesion and migration.
• Cress, A., Sroka, T. C., Pennington, M. E., & Cress, A. E. (2006). Synthetic D-amino acid peptide inhibits tumor cell motility on laminin-5. Carcinogenesis, 27(9).
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  Cell motility is partially dependent on interactions between the integrins and the extracellular matrix. Our previous studies have identified synthetic D-amino acid cell adhesion peptides using a combinatorial screening approach. In this study, we demonstrate that HYD1 (kikmviswkg) completely blocks random haptotactic migration and inhibits invasion of prostate carcinoma cells on laminin-5. This effect is adhesion independent and reversible. The inhibition of migration by HYD1 involves a dramatic remodeling of the actin cytoskeleton resulting in increased stress fiber formation and actin colocalization with cortactin at the cell membrane. HYD1 interacts with alpha6beta1 (not alpha6beta4) and alpha3beta1 integrins and surprisingly elevates laminin-5-dependent intracellular signals including focal adhesion kinase, mitogen-activated protein kinase kinase and extracellular signal-regulated kinase. HYD1 does not contain a previously characterized binding sequence for integrins. A scrambled derivative of HYD1, called HYDS (wiksmkivkg), does not interact with the alpha6 or alpha3 integrin subunits and is not biologically active. Taken together, these results indicate that HYD1 is a biologically active integrin-targeting peptide that reversibly inhibits tumor cell migration on laminin-5 and uncouples phosphotyrosine signaling from cytoskeletal-dependent migration.
• De Clerck, Y. A., Weissman, B. E., Yu, D., Parsons, R., Bar-Eli, M., Roy-Burman, P., Seewaldt, V. L., Cress, A. E., Languino, L. R., Batra, S. K., Tang, C. K., Sheng, S., Chen, W. T., Chellappan, S., Cheng, S. Y., Ladisch, S., McCarthy, J. B., Coussens, L. M., & Cohen, M. B. (2006). Tumor progression and metastasis from genetic to microenvironmental determinants: a workshop of the tumor progression and metastasis NIH study section in honor of Dr. Martin L. Padarathsingh, May 31, 2006, Georgetown, Washington, DC. Cancer biology & therapy, 5(12), 1588-99.
• De, C., Weissman, B. E., Yu, D., Parsons, R., Bar-Eli, M., Roy-Burman, P., Seewaldt, V. L., Cress, A. E., Languino, L. R., Batra, S. K., Tang, C. K., Sheng, S., Chen, W., Chellappan, S., Cheng, S., Ladisch, S., McCarthy, J. B., Coussens, L. M., & Cohen, M. B. (2006). Tumor progression and metastasis from genetic to microenvironmental determinants - A workshop of the tumor progression and metastasis NIH study section in honor of Dr. Martin L. Padarathsingh, May 31, 2006, Georgetown, Washington, DC. CANCER BIOLOGY & THERAPY, 5(12), 1588-1599.
• Sroka, T. C., Marik, J., Pennington, M. E., Lam, K. S., & Cress, A. E. (2006). The minimum element of a synthetic peptide required to block prostate tumor cell migration. CANCER BIOLOGY & THERAPY, 5(11), 1556-1562.
• Sroka, T. C., Pennington, M. E., & Cress, A. E. (2006). Synthetic D-amino acid peptide inhibits tumor cell motility on laminin-5. CARCINOGENESIS, 27(9), 1748-1757.
• Bair, E. L., Chen, M. L., McDaniel, K., Sekiguchi, K., Cress, A. E., Nagle, R. B., & Bowden, G. T. (2005). Membrane type 1 matrix metalloprotease cleaves laminin-10 and promotes prostate cancer cell migration. Neoplasia (New York, N.Y.), 7(4), 380-9.
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  Disruption of the extracellular matrix by proteases is crucial for tumor invasion. Laminin-10 (Ln-10) has previously been identified as a substrate for cell migration and cell adhesion, and is present in the basal lamina (BL) of both normal prostate and prostate cancer. Here, we investigate a role for membrane type 1 matrix metalloprotease (MT1-MMP) in modifying this Ln-10-rich BL. MT1-MMP is a transmembrane member of the MMP family that has been demonstrated to be upregulated as prostate cancer progresses from normal to prostate intraepithelial neoplasia to invasive cancer, suggesting a role for MT1-MMP in the invasion of prostate cancer. We show that MT1-MMP cleaves the alpha5 chain of purified human Ln-10 from its 350-kDa form into 310-, 190-, 160-, and 45-kDa fragments. This cleavage causes a decrease in DU-145 prostate cancer cell adhesion to purified Ln-10, and an increase in transmigration of DU-145 cells through cleaved Ln-10. We also show that prostate cancer cells expressing membrane-bound MT1-MMP cleave the alpha5 chain of Ln-10. Ln alpha5-chain cleavage is also observed in human prostate cancer tissues. These findings suggest that prostate cancer cells expressing high levels of MT1-MMP have increased invasive potential through their ability to degrade and invade Ln-10 barriers.
• Cress, A. E. (2005). Epigenetic regulation of the cell type-specific gene 14-3-3sigma.. Neoplasia (New York, N.Y.).
• Cress, A. E. (2005). Epigenetic silencing of DSC3 is a common event in human breast cancer.. Breast cancer research : BCR.
• Cress, A. E. (2005). Identification of a stem cell candidate in the normal human prostate gland.. European journal of cell biology.
• Cress, A. E. (2005). Membrane type 1 matrix metalloprotease cleaves laminin-10 and promotes prostate cancer cell migration.. Neoplasia (New York, N.Y.).
• Cress, A., Oshiro, M. M., Futscher, B. W., Lisberg, A., Wozniak, R. J., Klimecki, W. T., Domann, F. E., & Cress, A. E. (2005). Epigenetic regulation of the cell type-specific gene 14-3-3sigma. Neoplasia (New York, N.Y.), 7(9).
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  Epigenetic control participates in processes crucial in mammalian development, such as X-chromosome inactivation, gene imprinting, and cell type-specific gene expression. We provide evidence that the p53-inducible gene 14-3-3sigma is a new example of a gene important to human cancer, where epigenetic mechanisms participate in the control of normal cell type-specific expression, as well as aberrant gene silencing in cancer cells. Like a previously identified cell type-specific gene maspin, 14-3-3sigma is a p53-inducible gene; however, it participates in G2/M arrest in response to DNA-damaging agents. 14-3-3Sigma expression is restricted to certain epithelial cell types, including breast and prostate, whereas expression is absent in nonepithelial tissues such as fibroblasts and lymphocytes. In this report, we show that in normal cells expressing 14-3-3sigma, the 14-3-3sigma CpG island is unmethylated; associated with acetylated histones, unmethylated histone H3 lysine 9; and an accessible chromatin structure. By contrast, normal cells that do not express 14-3-3sigma have a methylated 14-3-3sigma CpG island with hypoacetylated histones, methylated histone H3 lysine 9, and an inaccessible chromatin structure. These findings extend the spectrum of cell type-specific genes controlled, partly, by normal epigenetic mechanisms, and suggest that this subset of genes may represent important targets of epigenetic dysregulation in human cancer.
• Oshiro, M. M., Kim, C. J., Wozniak, R. J., Junk, D. J., Muñoz-Rodríguez, J. L., Burr, J. A., Fitzgerald, M., Pawar, S. C., Cress, A. E., Domann, F. E., & Futscher, B. W. (2005). Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast cancer research : BCR, 7(5), R669-80.
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  Desmocollin 3 (DSC3) is a member of the cadherin superfamily of calcium-dependent cell adhesion molecules and a principle component of desmosomes. Desmosomal proteins such as DSC3 are integral to the maintenance of tissue architecture and the loss of these components leads to a lack of adhesion and a gain of cellular mobility. DSC3 expression is down-regulated in breast cancer cell lines and primary breast tumors; however, the loss of DSC3 is not due to gene deletion or gross rearrangement of the gene. In this study, we examined the prevalence of epigenetic silencing of DSC3 gene expression in primary breast tumor specimens.
• Schmelz, M., Moll, R., Hesse, U., Prasad, A. R., Gandolfi, J. A., Hasan, S. R., Bartholdi, M., & Cress, A. E. (2005). Identification of a stem cell candidate in the normal human prostate gland. European journal of cell biology, 84(2-3), 341-54.
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  Stem cells of the human prostate gland have not yet been identified utilizing a structural biomarker. We have discovered a new prostatic epithelial cell phenotype-expressing cytokeratin 6a (Ck6a+ cells). The Ck6a+ cells are present within a specialized niche in the basal cell compartment in fetal, juvenile and adult prostate tissue, and within the stem cell-enriched urogenital sinus. In adult normal prostate tissue, the average abundance of Ck6a+ cells was 4.9%. With proliferative stimuli in the prostate organ culture model, in which the epithelial-stromal interaction was maintained, a remarkable increase of Ck6a expression was noticed to up to 64.9%. The difference in cytokeratin 6a expression between the normal adult prostate and the prostate organ culture model was statistically significant (p
• Chauhan, S., Kunz, S., Davis, K., Roberts, J., Martin, G., Demetriou, M. C., Sroka, T. C., Cress, A. E., & Miesfeld, R. L. (2004). Androgen control of cell proliferation and cytoskeletal reorganization in human fibrosarcoma cells: role of RhoB signaling. The Journal of biological chemistry, 279(2), 937-44.
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  We recently generated an HT-1080-derived cell line called HT-AR1 that responds to dihydrotestosterone (DHT) treatment by undergoing cell growth arrest in association with cytoskeletal reorganization and induction of neuroendocrine-like cell differentiation. In this report, we show that DHT induces a dose-dependent increase in G0/G1 growth-arrested cells using physiological levels of hormone. The arrested cells increase in cell size and contain a dramatic redistribution of desmoplakin, keratin 5, and chromogranin A proteins. DHT-induced cytoskeletal changes were also apparent from time lapse video microscopy that showed that androgen treatment resulted in the rapid appearance of neuronal-like membrane extensions. Expression profiling analysis using RNA isolated from DHT-treated HT-AR1 cells revealed that androgen receptor activation leads to the coordinate expression of numerous cell signaling genes including RhoB, PTGF-beta, caveolin-2, Egr-1, myosin 1B, and EHM2. Because RhoB has been shown to have a role in tumor suppression and neuronal differentiation in other cell types, we investigated RhoB signaling functions in the HT-AR1 steroid response. We found that steroid induction of RhoB was DHT-specific and that newly synthesized RhoB protein was post-translationally modified and localized to endocytic vesicles. Moreover, treatment with a farnesyl transferase inhibitor reduced DHT-dependent growth arrest, suggesting that prenylated RhoB might function to inhibit HT-AR1 cell proliferation. This was directly shown by transfecting HT-AR1 cells with RhoB coding sequences containing activating or dominant negative mutations.
• Cress, A. E. (2004). Extracellular alpha 6 integrin cleavage by urokinase-type plasminogen activator in human prostate cancer.. Experimental cell research.
• Cress, A. E. (2004). Therapeutic targeting of prostate cancer. Cancer Biology & Therapy.
• Cress, A. E., & Mohla, S. (2004). Therapeutic targeting of prostate cancer. Cancer biology & therapy, 3(10), 1028-30.
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  An inaugural conference in Tucson Arizona on May 6-9, 2004 brought together more than 70 clinical and basic scientists to discuss recent research advances in understanding and targeting the progression of the human prostate cancer. The informal meeting was unique in that it provided the opportunity for discussion and interaction between these different groups of scientists whose paths rarely cross. The goal of the meeting was to develop new and novel approaches in understanding the human prostate cancer in order to uncover therapeutic targets. Faculty from six different cancer centers were represented including Memorial Sloan-Kettering Cancer Center (New York, NY); Arizona Cancer Center (Tucson, AZ); Fred Hutchinson Cancer Center (Seattle, WA); Chao Family Comprehensive Cancer Center (Irvine, CA); the Sydney Kimmel Cancer Center (San Diego, CA); Jonsson Comprehensive Cancer Center, University of California (Los Angeles, CA); and University of Massachusetts Memorial Cancer Center (Worcester, MA). Several important concepts emerged from this meeting as a result of the basic and clinical science interface. These concepts include: (1) Human prostate cancer has unique biological features as compared to other human epithelial malignancies; (2) Tumor plasticity is evident early in prostate cancer progression as evidenced by alterations in the extracellular matrix; (3) New therapeutic strategies should include the co-targeting of the stroma and prostate cancer; (4) Cell-cell and cell-ECM adhesion switching are reversible phenotypes evident early in human prostate tumor progression; (5) The discovery of molecular signatures including genomic or proteomic patterns for the discrimination of indolent versus aggressive disease is a potentially powerful tool and requires multifactorial approaches for success; and (6) New biomarkers and innovative tissue specific imaging modalities for human prostate cancer are being developed that may aid in a more accurate assessment of prostate cancer in patients.
• Cress, A., Demetriou, M. C., Pennington, M. E., Nagle, R. B., & Cress, A. E. (2004). Extracellular alpha 6 integrin cleavage by urokinase-type plasminogen activator in human prostate cancer. Experimental cell research, 294(2).
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  During human prostate cancer progression, the integrin alpha6beta1 (laminin receptor) is expressed on the cancer cell surface during invasion and in lymph node metastases. We previously identified a novel structural variant of the alpha6 integrin called alpha6p. This variant was produced on the cell surface and was missing the beta-barrel extracellular domain. Using several different concentrations of amiloride, aminobenzamidine and PAI-1 and the urokinase-type plasminogen activator (uPA) function-blocking antibody (3689), we showed that uPA, acting as a protease, is responsible for production of alpha6p. We also showed that addition of uPA in the culture media of cells that do not produce alpha6p, resulted in a dose-dependent alpha6p production. In contrast, the addition of uPA did not result in the cleavage of other integrins. Using alpha2-antiplasmin and plasmin depleted media, we observed that uPA cleaves the alpha6 integrin directly. Further, 12-o-tetradecanoyl-phorbol-13-acetate (TPA) induced the production of alpha6p, and this induction was abolished by PAI-1 but not alpha2-antiplasmin. Finally, the alpha6p integrin variant was detected in invasive human prostate carcinoma tissue indicating that this is not a tissue culture phenomenon. These data, taken together, suggest that this is a novel function of uPA, that is, to remove the beta-barrel ligand-binding domain of the integrin while preserving its heterodimer association.
• Demetriou, M. C., & Cress, A. E. (2004). Integrin clipping: a novel adhesion switch?. Journal of cellular biochemistry, 91(1), 26-35.
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  During human prostate cancer progression, the majority of normally expressed integrins are suppressed with the exception of the alpha6, alpha3, and beta1 integrins. We have shown that in prostate cancer, the alpha6 integrin is found paired with the beta1 integrin and that a novel form of the alpha6 integrin that lacks a large portion of the extracellular domain (alpha6p) exists. The alpha6pbeta1 integrin is found in human prostate cancer tissue specimens as well as tissue culture cell lines and is formed on the cell surface. This review discusses the mechanism of alpha6pbeta1 production and the potential functions of this integrin variant. Our current working model predicts that the alpha6pbeta1 integrin maintains the intracellular cytoskeletal connections associated with the heterodimer while allowing for an alteration in cell adhesion. The mechanism provides a selective advantage for cancer cell metastasis.
• Chauhan, S., Pandey, R., Way, J. F., Sroka, T. C., Demetriou, M. C., Kunz, S., Cress, A. E., Mount, D. W., & Miesfeld, R. L. (2003). Androgen regulation of the human FERM domain encoding gene EHM2 in a cell model of steroid-induced differentiation. Biochemical and biophysical research communications, 310(2), 421-32.
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  We have developed a cell model to investigate steroid control of differentiation using a subline of HT1080 cells (HT-AR1) that have been engineered to express the human androgen receptor. Dihydrotestosterone (DHT) treatment of HT-AR1 cells induced growth arrest and cytoskeletal reorganization that was associated with the expression of fibronectin and the neuroendocrine markers chromogranin A and neuron-specific enolase. Expression profiling analysis identified the human FERM domain-encoding gene EHM2 as uniquely induced in HT-AR1 cells as compared to 16 other FERM domain containing genes. Since FERM domain proteins control cytoskeletal functions in differentiating cells, and the human EHM2 gene has not been characterized, we investigated EHM2 steroid-regulation, genomic organization, and sequence conservation. We found that DHT, but not dexamethasone, induced the expression of a 3.8 kb transcript in HT-AR1 cells encoding a 504 amino acid protein, and moreover, that human brain tissue contains a 5.8 kb transcript encoding a 913 amino acid isoform. Construction of an unrooted phylogenetic tree using 98 FERM domain proteins revealed that the human EHM2 gene is a member of a distinct subfamily consisting of nine members, all of which contain a highly conserved 325 amino acid FERM domain.
• Cress, A. E. (2003). Androgen control of cell proliferation and cytoskeletal reorganization in human fibrosarcoma cells: role of RhoB signaling.. The Journal of biological chemistry.
• Cress, A. E. (2003). Androgen regulation of the human FERM domain encoding gene EHM2 in a cell model of steroid-induced differentiation.. Biochemical and biophysical research communications.
• Cress, A. E. (2003). Integrin clipping: A novel adhesion switch?. Journal of Cellular Biochemistry.
• Cress, A. E. (2003). Membrane type-1-matrix metalloproteinase expressed by prostate carcinoma cells cleaves human laminin-5 beta3 chain and induces cell migration.. Cancer research.
• Udayakumar, T. S., Chen, M. L., Bair, E. L., Von Bredow, D. C., Cress, A. E., Nagle, R. B., & Bowden, G. T. (2003). Membrane type-1-matrix metalloproteinase expressed by prostate carcinoma cells cleaves human laminin-5 beta3 chain and induces cell migration. Cancer research, 63(9), 2292-9.
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  Degradation of the extracellular matrix by proteolytic enzymes is a central aspect of physiological and pathologic tissue-remodeling processes such as trophoblastic implantation, wound healing, and tumor invasion. We have hypothesized that prostate adenocarcinoma cell invasion through the normal basal lamina is attributable in part to metalloproteinase-induced cleavage of laminin-5 (Ln-5) and enhanced motility of the cancer cells. We studied the role of membrane type-1-matrix metalloproteinase (MT1-MMP) expressed on the surface of prostate tumor cells in cleaving Ln-5 and enhancing the migration of prostate tumor cells. We also determined the nature of the MT1-MMP cleavage of human Ln-5 and how this altered Ln-5 changes the migration of prostate carcinoma cells. We found that human MT1-MMP cleaves purified human Ln-5 to an 80-kDa fragment. Mass spectrometry analyses of the 80-kDa cleaved product by trypsin and chymotrypsin gave 14 and 9 different peptide sequences, respectively, that were identical to the expected amino acid sequence of the Ln-5-beta3 chain. The recovered peptides represent 14.4% (trypsin) and 10.3% (chymotrypsin) of Ln-5-beta3 chain by amino acid count. Both trypsin and chymotrypsin digestion of MT1-MMP-cleaved product of Ln-5 did not show any other peptides that were identical to the other chains of Ln-5. Using a linear migration assay we found that the Ln-5 cleaved by MT1-MMP enhanced the migration of DU-145 prostate carcinoma cells by 2-fold compared with uncleaved Ln-5. The use of blocked antisense MT1-MMP oligonucleotides inhibited the migration of DU-145 cells on Ln-5. We also found that the prostate carcinoma cells expressing high levels of MT1-MMP, such as PC3N and PPC, demonstrated enhanced migration on human Ln-5-coated substrate, and this migration was inhibited using blocked antisense MT1-MMP oligonucleotides. In conclusion, this is a novel and important finding where we have shown that beta3-chain is cleaved by MT1-MMP, and this cleavage enhances migration of prostate cancer cells.
• Cress, A. E. (2002). Androgen induction of in vitro prostate cell differentiation.. Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.
• Cress, A. E. (2002). Culturing precision-cut human prostate slices as an in vitro model of prostate pathobiology.. Cell biology and toxicology.
• Cress, A. E. (2002). Different phenotypes in human prostate cancer: alpha6 or alpha3 integrin in cell-extracellular adhesion sites.. Neoplasia (New York, N.Y.).
• Cress, A. E. (2002). Differential regulation of a novel variant of the alpha(6) integrin, alpha(6p).. Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.
• Parrish, A. R., Sallam, K., Nyman, D. W., Orozco, J., Cress, A. E., Dalkin, B. L., Nagle, R. B., & Gandolfi, A. J. (2002). Culturing precision-cut human prostate slices as an in vitro model of prostate pathobiology. Cell biology and toxicology, 18(3), 205-19.
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  Due to the complex morphology of the prostate, it was hypothesized that precision-cut tissue slices from human prostate would provide a unique in vitro model. Precision-cut slices were generated from zones of human prostate and their viability was assessed under conditions of different media for up to 120 h. Slices were also exposed to several concentrations of CdCI2, which was used as a model toxicant. Maintenance of both stromal and epithelial cells was noted; however, there was a gradual loss of luminal epithelial cells when the medium was not supplemented with dihydrotestosterone (DHT). Minimal leakage of lactate dehydrogenase occurred throughout the incubation. Prostate-specific antigen (PSA) was detected in the medium at all time points, although the rates of secretion fell over time. There was a loss of PSA-positive cells when the medium was not supplemented with DHT, consistent with a loss of luminal cells, whereas PSA-positive cells were maintained in the DHT-supplemented media. A proliferation of basal cells was observed in the presence of media containing 10% fetal bovine serum. Exposure of slices to CdCl2 demonstrated a dose-response effect ranging from proliferation to complete cellular necrosis. Given the retention of stromal-epithelial interactions and the use of acquired human tissue, prostate slices represent a unique in vitro model for investigating human prostate pathobiology.
• Cress, A. E. (2001). Identification of a novel structural variant of the alpha 6 integrin.. The Journal of biological chemistry.
• Cress, A. E. (2001). Integrin- and cadherin-mediated induction of the matrix metalloprotease matrilysin in cocultures of malignant oral squamous cell carcinoma cells and dermal fibroblasts.. Experimental cell research.
• Cress, A. E. (2001). Laminin-5-mediated gene expression in human prostate carcinoma cells.. Molecular carcinogenesis.
• Cress, A. E. (2001). Synthetic peptides inhibit adhesion of human tumor cells to extracellular matrix proteins.. Cancer research.
• Cress, A. E. (2001). Unique expression pattern of the ?6?4 integrin and laminin-5 in human prostate carcinoma. The Prostate.
• Cress, A. E. (2001). Unique expression pattern of the α6β4 integrin and laminin‐5 in human prostate carcinoma. The Prostate.
• Cress, A. E. (2000). Altered surface expression and increased turnover of the alpha6beta4 integrin in an undifferentiated carcinoma.. Carcinogenesis.
• Cress, A. E. (2000). Quantitation of phosphotyrosine signals in human prostate cell adhesion sites.. BioTechniques.
• Cress, A. E. (1999). Cell adhesion mediated drug resistance (CAM-DR): role of integrins and resistance to apoptosis in human myeloma cell lines.. Blood.
• Cress, A. E. (1999). Multiple mechanisms confer drug resistance to mitoxantrone in the human 8226 myeloma cell line.. Cancer research.
• Cress, A. E. (1999). N-Cadherin Expression in Human Prostate Carcinoma Cell Lines. The American Journal of Pathology.
• Cress, A. E. (1998). Activation of gelatinase-tissue-inhibitors-of-metalloproteinase complexes by matrilysin.. The Biochemical journal.
• Cress, A. E. (1997). A human xenograft model for testing early events of epithelial neoplastic invasion.. International journal of oncology.
• Cress, A. E. (1997). Cleavage of beta 4 integrin by matrilysin.. Experimental cell research.
• Cress, A. E. (1997). Free radicals generated by ionizing radiation signal nuclear translocation of p53.. Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.
• Cress, A. E. (1996). Cytokeratin expression results in a drug-resistant phenotype to six different chemotherapeutic agents.. Clinical cancer research : an official journal of the American Association for Cancer Research.
• Cress, A. E. (1996). Differential expression of laminin 5 (alpha 3 beta 3 gamma 2) by human malignant and normal prostate.. The American journal of pathology.
• Cress, A. E. (1996). Evidence for cytoplasmic P-glycoprotein location associated with increased multidrug resistance and resistance to chemosensitizers.. Cancer research.
• Cress, A. E. (1996). Multiple drug resistance and intermediate filaments. Cancer and Metastasis Review.
• Cress, A. E. (1996). The use of a combinatorial library method to isolate human tumor cell adhesion peptides. Molecular Diversity.
• Nagle, R. B., Woo, C., Stea, B., Nagle, R. B., & Cress, A. E. (1996). 131 The integrin α6β4 as a signaling membrane protein for a damage response to ionizing radiation in human prostate cancer cell lines. International Journal of Radiation Oncology Biology Physics, 36(1), 224. doi:10.1016/s0360-3016(97)85473-9
• Cress, A. E. (1995). Degradation of fibronectin fibrils by matrilysin and characterization of the degradation products.. Experimental cell research.
• Cress, A. E. (1995). Expression of hemidesmosomal and extracellular matrix proteins by normal and malignant human prostate tissue.. The American journal of pathology.
• Cress, A. E. (1995). Integrin alpha 6 expression in human prostate carcinoma cells is associated with a migratory and invasive phenotype in vitro and in vivo.. Clinical & experimental metastasis.
• Cress, A. E. (1995). The ?6?1 and ?6?4 integrins in human prostate cancer progression. Cancer and Metastasis Reviews.
• Cress, A. E. (1994). Adhesion molecules, extracellular matrix, and proteases in prostate carcinoma.. Journal of cellular biochemistry. Supplement.
• Cress, A. E. (1994). Biosynthesis and secretion of laminin and S-laminin by human prostate carcinoma cell lines. The Prostate.
• Cress, A. E. (1994). Differential expression of extracellular matrix molecules and the alpha 6-integrins in the normal and neoplastic prostate.. The American journal of pathology.
• Cress, A. E. (1994). Expression of cytokeratin confers multiple drug resistance.. Proceedings of the National Academy of Sciences of the United States of America.
• Cress, A. E. (1994). Heat shock-induced shedding of cell surface integrins in A549 human lung tumor cells in culture.. Experimental cell research.
• Cress, A. E. (1994). Immunohistochemical analysis of cathepsin D in prostate carcinoma.. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc.
• Cress, A. E. (1994). Polyamine-dependent expression of the matrix metalloproteinase matrilysin in a human colon cancer-derived cell line.. Molecular carcinogenesis.
• Cress, A. E. (1993). A DNA polymerase alpha-associated 56 kDa protein kinase.. Biochemical and biophysical research communications.
• Cress, A. E. (1993). Characterization of integrin subunits, cellular adhesion and tumorgenicity of four human prostate cell lines.. Journal of cancer research and clinical oncology.
• Cress, A. E. (1992). Alteration of human tumor cell adhesion by high-strength static magnetic fields.. Investigative radiology.
• Cress, A. E. (1992). Delta-type DNA polymerase characterized from Drosophila melanogaster embryos.. Nucleic acids research.
• Cress, A. E. (1992). New relationships between prostatic intraepithelial neoplasia and prostatic carcinoma. Journal of Cellular Biochemistry.
• Cress, A. E. (1992). New relationships between prostatic intraepithelial neoplasia and prostatic carcinoma.. Journal of cellular biochemistry. Supplement.
• Cress, A. E. (1990). Alteration of cellular adhesion by heat shock. Experimental Cell Research.
• Cress, A. E. (1990). Profiles of human melanoma cell surface proteins: effects of culturing on two different substrates.. Pigment cell research.
• Cress, A. E. (1990). The crosslinking of nuclear protein to DNA using ionizing radiation.. Journal of cancer research and clinical oncology.
• Nagle, R. B., Majda, J. A., Gehlsen, K. R., & Cress, A. E. (1990). Detection of human tumor epithelial micrometastases in bone marrow using antibodies to acidic keratins and integrin α3β1.(Project supported by a 1989–1990 ASTRO fellowship grant). International Journal of Radiation Oncology Biology Physics, 19, 211. doi:10.1016/0360-3016(90)90816-3
• Cress, A. E. (1989). Activation of the cellular Harvey ras gene in mouse skin tumors initiated with urethane.. Molecular carcinogenesis.
• Cress, A. E. (1989). Expression of β-actin during progression of mouse skin tumors. Carcinogenesis.
• Cress, A. E. (1989). Nuclear protein organization and the repair of radiation damage.. Carcinogenesis.
• Cress, A. E. (1989). Persistent intracellular binding of mitoxantrone in a human colon carcinoma cell line. Biochemical Pharmacology.
• Cress, A. E. (1988). Cytogenetic and phenotypic analysis of a human colon carcinoma cell line resistant to mitoxantrone.. Cancer research.
• Cress, A. E. (1988). Identification of attachment proteins for DNA in Chinese hamster ovary cells.. The Journal of biological chemistry.
• Cress, A. E. (1988). Modification of keratin by the chemotherapeutic drug mitoxantrone. Biochemical Pharmacology.
• Cress, A. E. (1987). An analysis of DNA replication in synchronized CHO cells treated with benzo[a]pyrene diol epoxide.. Biochimica et biophysica acta.
• Cress, A. E. (1986). Characterization of a new drug-resistant human myeloma cell line that expresses P-glycoprotein.. Cancer research.
• Cress, A. E. (1985). Rapid loss of stress fibers in Chinese hamster ovary cells after hyperthermia.. Cancer research.
• Cress, A. E. (1983). Covalent DNA-Protein Crosslinking Occurs after Hyperthermia and Radiation. Radiation Research.
• Cress, A. E. (1983). The effect of heat and radiation on the initiation and elongation processes of DNA synthesis.. International journal of radiation biology and related studies in physics, chemistry, and medicine.
• Cress, A. E. (1982). Correlation between amounts of cellular membrane components and sensitivity to hyperthermia in a variety of mammalian cell lines in culture.. Cancer research.
• Cress, A. E. (1982). Thermal Enhancement of X-Ray-Induced DNA Crosslinking. Radiation Research.
• Cress, A. E. (1981). PH stepwise alkaline elution of DNA replication intermediates during S phase. Biochemical and Biophysical Research Communications.
• Cress, A. E. (1981). Reversal of resistance to methotrexate by hyperthermia in Chinese hamster ovary cells.. Cancer research.
• Cress, A. E. (1980). Cholesterol levels inversely reflect the thermal sensitivity of mammalian cells in culture.. Nature.
• Cress, A. E. (1980). Factors regulating membrane permeability alter thermal resistance.. Annals of the New York Academy of Sciences.
• Cress, A. E. (1980). Ornithine decarboxylase induction in cells stimulated to proliferate differs from that in continuously dividing cells.. The Biochemical journal.
• Cress, A. E. (1979). Collateral sensitivity to methotrexate in cells resistant to adriamycin.. Cancer research.
• Cress, A. E. (1979). Hydroxyurea inhibits ODC induction, but not the G1 to S phase transition. Biochemical and Biophysical Research Communications.
• Cress, A. E. (1977). Hydroxyurea treatment affects the G1 phase in next generation CHO cells.. Experimental cell research.

Proceedings Publications

• Jacobson, J. R., Garcia, J. G., Epshtein, Y., Cress, A. E., & Chen, W. (2021). Human Lung Endothelial Cell Barrier Function Is Augmented by Kindlin 2. In TP113. TP113 ACUTE LUNG INJURY AND REPAIR.
• Sun, X., Sun, B., Oita, R. C., Garcia, J. G., Gallegos, P., Cress, A. E., & Angle, D. (2020). Transcription Regulation of NAMPT by Radiation in Human Lung Endothelium and Epithelium. In C58. ACUTE LUNG INJURY: ATYPICAL TRIGGERS.

Poster Presentations

• Warfel, N. A., Chauhan, S. C., Snider, J., Snider, A. J., & Cress, A. E. (2022, winter).

  PIM1 derives lipid droplet accumulation via regulation of GSK3β-PPARα signaling axis to promote proliferation and survival in prostate cancer

  . FASEB lipid droplet conferenceFASEB.

Others

• Cress, A. E. (2019). EPITHELIAL CELL SURFACE TARGETING USING SYNTHETIC D-AMINO ACID PEPTIDES. Cell Adhesion and Cytoskeletal Molecules in Metastasis.
• Cress, A. E. (2019). SUPPRESSION AND ALTERATION OF ADHESION STRUCTURES IN HUMAN EPITHELIAL CANCER PROGRESSION. Cell Adhesion and Cytoskeletal Molecules in Metastasis.
• Cress, A. E. (2018, June). Androgen Receptor-Induced Integrin α6β1 and Adhesion to Laminin Promotes Survival and Drug Resistance in Castration-Resistant Prostate Cancer through BNIP3.
• Cress, A. E. (2013, September). Prevention of Prostate Cancer. Fundamentals of Cancer Prevention.