Anne E Cress
- Deputy Dean, Research Affairs
- Professor, Cellular and Molecular Medicine
- Professor, Radiation Oncology
- Professor, Molecular and Cellular Biology
- Professor, Applied BioSciences - GIDP
- Professor, Cancer Biology - GIDP
OMB No. 0925-0001/0002 (Rev. 08/12 Approved Through 8/31/2015)
Provide the following information for the Senior/key personnel and other significant contributors.
Follow this format for each person. DO NOT EXCEED FIVE PAGES.
NAME: Anne E. Cress, PhD
POSITION TITLE: Professor of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ
eRA COMMONS USER NAME (credential, e.g., agency login): acress
EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and residency training if applicable. Add/delete rows as necessary.)
INSTITUTION AND LOCATION
FIELD OF STUDY
University of Arizona, Tucson, AZ
University of Arizona, Tucson, AZ
University of Arizona, Tucson, AZ
Stanford University, Stanford, CA
University of Queensland, Brisbane, Australia
Netherlands Cancer Institute, Amsterdam
A. Personal Statement. My lab is dedicated to elucidating the molecular mechanisms of human cancer progression and metastasis. Specifically, we study the regulation of cell surface molecules (called integrins) and their role in cancer cell adhesion to the extracellular matrix. Among our many contributions, my research team discovered that laminin adhesion structures are dramatically altered in human cancer resulting in invasion, metastasis, and drug resistance. Additionally, my lab has shown that the laminin binding integrins, (A6B1, A3B1, and A6B4 integrin) can be targeted to prevent cancer progression and metastasis by inducing dormant disease. We have contributed many publications in this area and developed three approaches to interrupt cell adhesion to laminin: (1) using cyclized peptides, (2) deploying small molecules, and/or (3) using a function-blocking antibody. I spent a sabbatical period working at a drug development and design institute in Australia to screen and design peptide ligand mimetics. In a second sabbatical, I worked at the Netherlands Cancer Institute to screen for anti-integrin antibodies in the laboratory of Dr. Arnoud Sonnenberg, who discovered the A6B4 integrin and continues to work on its regulation. As both an educator and researcher, my work is characterized by a team-based approach using experts in human oncology, cell biology, biochemistry, and pharmacology to propose and test innovative solutions to address unmet clinical needs.
B. Positions and Honors
1980-1981 Research Associate, Division of Radiation Oncology, University of Arizona, Tucson, AZ
1981-1985 Research Assistant Professor, Division of Radiation Oncology, University of Arizona, Tucson, AZ
1985-1989 Assistant Professor, Radiation Oncology, University of Arizona, Tucson, AZ
1990-1996 Associate Professor, Radiation Oncology, University of Arizona, Tucson, AZ
1996- Professor, Radiation Oncology, University of Arizona, Tucson, AZ
1999- Professor, Molecular and Cellular Biology, University of Arizona, Tucson, AZ
1999 Outstanding Research Award, H. Lee Moffitt Cancer Center, Tampa, FL
2003-2009 Associate Dean for Research, College of Medicine, University of Arizona, Tucson, AZ
2003- Professor, Cellular and Molecular Medicine, University of Arizona, Tucson, AZ
2005 Elkin Award for Cancer Biology Research, Emory University
2005 Founders Day Award, University of Arizona College of Medicine
2008 Sydney E. Salmon MD Award for Excellence in Research, University of Arizona
2010- Deputy Dean for Research, College of Medicine, University of Arizona, Tucson, AZ
2013-2014 Interim Director, UA Cancer Center, Tucson, AZ
2015- Interim Associate Director for Basic Sciences, UA Cancer Center, Tucson, AZ
NIH Activities and Memberships:
1980- Member, American Association of Cancer Research (AACR)
1990- Member, American Society for Cell Biology
2005- Member, Metastasis Research Society
1997-2001 American Cancer Society, Chair, Peer Review Committee on Cell Cycle and Growth Control
1998-2003 NIH, Member and Chairperson, Radiation Research Study Section
2003- NIH, Special Emphasis Panel Grant Reviews, Cancer Center Core Grant Reviews
2004-2010 NIH, Member, Tumor Progression and Metastasis (TPM) Study Section
2008-2010 NIH, Chairperson, TPM Study Section
2012-2014 American Association for Cancer Research, National Millennium Award Review Committee
2015- NIH, Reviewer, F09-B Oncology Fellowship Panel, F31, F32 Grants
C. Contribution to Science
1. Discovery of cell adhesion mediated drug resistance. Our group discovered Cell Adhesion Mediated Drug Resistance (CAM-DR), which we have shown to be a significant impediment to tumor eradication. Normal and tumor cells respond to DNA damage caused by ionizing radiation (IR) and chemotherapeutic agents. Epithelial cells are resistant to lethal effects of DNA damaging agents dependent upon cytokeratin. Adhesion to laminin 5, the ligand for ITGA6, promotes a G2 progression block in normal cells in response to IR. Without laminin 5 adhesion, the DNA damage-induced block was significantly reduced. The working hypothesis is that laminin 5 adhesion, and the structural integrin-cytokeratin connections to the nucleus, promote a robust DNA damage response. We speculate that defective laminin adhesion in early cancer progression has an unintended consequence of promoting genomic instability via attenuation of the DNA damage response.
a. Expression of cytokeratin confers multiple drug resistance, PA Bauman, WS Dalton, JM Anderson, AE Cress. Proceedings of the National Academy of Sciences 91 (12), 5311-5314. PMCID: PMC43984
b. Cell adhesion mediated drug resistance (CAM-DR): role of integrins and resistance to apoptosis in human myeloma cell lines JS Damiano, AE Cress, LA Hazlehurst, AA Shtil, WS Dalton. Blood 93 (5), 1658-1667. PMID: 10029595
c. Cytokeratin expression results in a drug-resistant phenotype to six different chemotherapeutic agents. JM Anderson, LM Heindl, PA Bauman, CW Ludi, WS Dalton, AE Cress. Clinical Cancer Research 2 (1), 97-105. PMID: 9816096
d. Gemcitabine resistant pancreatic cancer cell lines acquire an invasive phenotype with collateral hypersensitivity to histone deacetylase inhibitors. Samulitis BK, Pond KW, Pond E, Cress AE, Patel H, Wisner L, Patel C, Dorr RT, Landowski TH. Cancer Biol Ther. 2015;16(1):43-51. doi: 10.4161/15384047.2014.986967.
2. Switching adhesion complexes is an early event in epithelial cancer progression. The regulator of normal skin and glandular tissue homeostasis is the A6B4 integrin. It is a laminin receptor that acts as a seed site for the formation of hemidesomsome (HD) structures. In the skin and corneal epithelium, human blistering diseases result from germline mutations in either the receptor or the ligand. We discovered that HDs and focal adhesions (FAs) are present in normal prostate glands. Further, we found that the HD structure is attenuated in intraepithelial neoplasia and, in invasive and metastatic human prostate cancer, is switched to A6B1 integrin. Loss of the HD structure is correlated with later stage and higher grade of the disease. Preventing the switching of the adhesion structure is an actionable step in preventing the metastatic spread of this slow growing type cancer.
a. Integrin α6 expression in human prostate carcinoma cells is associated with a migratory and invasive phenotype in vitro and in vivo. I Rabinovitz, RB Nagle, AE Cress. Clinical & experimental metastasis 13 (6), 481-491. PMCID: PMC2846819
b. Identification of a stem cell candidate in the normal human prostate gland. M Schmelz, R Moll, U Hesse, AR Prasad, JA Gandolfi, SR Hasan, Cress, AE. European J of Cell Biology 84 (2), 341-354. PMCID: PMC2730953
c. Schwann cells increase prostate and pancreatic tumor cell invasion on laminin. Isis C. Sroka, Hasharon Chopra, Lipsa Das, Jaime M.C. Gard, Raymond B. Nagleand Anne E. Cress. J Cell Biochem. 2015 Aug 3. doi: 10.1002/jcb.25300. [Epub ahead of print] PMID: 26239765
3. Novel tumor specific forms of laminin binding integrins promote invasion and metastasis. We discovered structural variant forms of ITGA6 and ITGB4 that occur specific to cancer progression. A novel post-translational modification (PTM) of ITGA6 generated a novel form of ITGA6, called ITGA6p, produced on the cancer cell surface by the protease urokinase. Infiltrating macrophages into the tumor promote the production of ITGA6p. The urokinase plasminogen activator receptor (uPAR, PLAUR) is required for the cleavage of ITGA6 by uPA. The working hypothesis is that ITGA6p dominates in cohesive migration and establishes cancer residency in metastatic sites such as the bone.
a. Identification of a novel structural variant of the α6 integrin. TL Davis, I Rabinovitz, BW Futscher, M Schnölzer, F Burger, Y Liu, Cress, AE. Journal of Biological Chemistry 276 (28), 26099-26106. PMCID: PMC2824502
b. Extracellular Engagement of α6 Integrin Inhibited Urokinase-Type Plasminogen Activator–Mediated Cleavage and Delayed Human Prostate Bone Metastasis. MO Ports, RB Nagle, GD Pond, AE Cress. Cancer Research 69 (12), 5007-5014. PMCID: PMC2697270
c. Laminin-binding integrin gene copy number alterations in distinct epithelial-type cancers.
William L. Harryman, Erika Pond, Parminder Singh, Andrew S. Little, Jennifer M. Eschbacher, Raymond B. Nagle and Anne E. Cress. Amer J Transl Res. 2016, accepted for publication.
4. Preventing bone metastasis progression and cancer pain. Our goal is to understand human prostate tumor cell adhesion and migration, and use this information to block subsequent aggressive spreadto late stage secondary skeletal sites. Early (and often clinically non-apparent) bone metastasis can provide a sanctuary site for tumor cells and result in widespread skeletal recurrence and pain resulting from aggressive metastatic disease 5 to 10 years after primary therapy. We discovered a new therapeutic strategy and concept of cancer control directed at blocking the success of early adhesion dependent tumors and in particular, painful bone-resident cancer.
a. The role of alpha 6 integrin in prostate cancer migration and bone pain in a novel xenograft model.
King TE, Pawar SC, Majuta L, Sroka IC, Wynn D, Demetriou MC, Nagle RB, Porreca F, Cress AE.
PLoS One. 2008;3(10):e3535. doi: 10.1371/journal.pone.0003535. Epub 2008 Oct 28. PMCID: PMC2570216
b. Inhibition of p38-MAPK signaling pathway attenuates breast cancer induced bone pain and disease progression in a murine model of cancer-induced bone pain. Sukhtankar D, Okun A, Chandramouli A, Nelson MA, Vanderah TW, Cress AE, Porreca F, King T. Mol Pain. 2011 Oct 20;7:81. doi: 10.1186/1744- 8069-7-81. PMCID: PMC3212934
c. Targeting integrin α6 stimulates curative-type bone metastasis lesions in a xenograft model.
Landowski TH, Gard J, Pond E, Pond GD, Nagle RB, Geffre CP, Cress AE. Mol Cancer Ther. 2014 Jun;13(6):1558-66. doi: 10.1158/1535-7163.MCT-13-0962. Epub 2014 Apr 16. PMCID: PMC4069206
5. Training and mentoring approximately 49 students at all levels for professional careers in the medical and biological sciences and biotechnology. Curiosity coupled with quantitative skills is a rare combination in students that I have had the honor to nurture. I am proud that the majority of past students are now either current or past Cancer Center Directors, Professors at major U.S. Universities, leaders in biotechnology, or full partners in prestigious patent law firms. All of these students were very intelligent, highly motivated, and perhaps most important, persistent in their pursuit of excellence. I view my contribution to their career path as a valuable addition to science that could not have been accomplished without my own scientific curiosity and pursuit of peer-reviewed biomedical research.
1. #6,812,003: Compounds and Methods for Modulating Cell-Adhesion Mediated Drug Resistance.
Peptides and methods of their use for inhibiting drug and radiation therapy resistance in cancerous cells in which the efficacy of chemotherapy and/or radiation therapy is enhanced by administration of an effective about of a peptide that inhibits cell adhesion mediated drug resistance (CAM-DR). The peptide is preferably administered to the patient prior to chemotherapy or radiation therapy. Inhibition of CAM-DR by RZ-3 in multiple myeloma is disclosed.
2. #7,253,149: HYD1 peptides as anti-cancer agents in cancer.
Complete List of Published Work in My Bibliography:
http://www.ncbi.nlm.nih.gov/sites/myncbi/anne.cress.1/bibliography/41520311/public/?sort=date&direction=descending More than 183 peer-reviewed publications and 4 book chapters with a Google Scholar h-index of 38.
D. Research Support
5R01 CA159406 (PI: Cress) 12/11/2011 – 05/31/2016 1.32 CM
Human Prostate Cancer Metastasis and Laminin Binding Integrins
The objective is to understand human prostate tumor cell adhesion, migration, and to use this information to block subsequent spread to late stage secondary skeletal sites. We will identify the role of the laminin binding integrins A6B1, A6pB1, and A3B1 at the molecular and cellular level using both in vitro tissue culture and in vivo xenograft SCID mouse models. The efficacy of specific antibodies and small molecule reagents to prevent bone metastasis will be tested.
3P30CA023074 (PI: Kraft, PI) 08/19/2009 – 06/30/2016 0.6 CM
Arizona Cancer Center – Cancer Center Support Grant
The University of Arizona Cancer Center (UACC) has been a highly productive NCI-designated Comprehensive Cancer Center for nearly 40 years. This Cancer Center Support Grant provides support for the critical infrastructure and shared resources needed by our scientific program investigators to seek new treatment methods to both prevent and cure cancer.
Role: Co-investigator and Associate Director for Research
NCI T32CA09213 (PI: Martinez) 05/19/2015 - 04/30/2020 No salary support
Cancer Biology Training Grant $321,281/yr
The objective of this T32 training grant is to identify talented pre- and post-doctoral students and develop outstanding cancer biology researchers by placement in cancer biology laboratories that are conducting cutting edge research, development of a thorough cancer biology knowledge base with a thoughtfully constructed lecture schedule, and through skilled mentoring and evaluation.
Role: Faculty Mentor
P01 HL126609 (PD/PI: Garcia) 04/01/2016 – 03/31/2021 2.4 CM
NIH/NHLBI $1,660,188/yr DC
Cytoskeletal Regulation of Lung Endothelial Pathobiology
Project #3: Integrin b4 and Paxillin in EC Focal Adhesion Dynamics and Barrier Responses
In its 16th-20th year of proposed funding, the PPG will investigate the complex field of inflammatory lung injury, particularly, the spatial regulation of the dynamic actomyosin cytoskeleton (central stress fibers, lamellipodia formation, focal adhesion formation) involving MLCK, cortactin, c-Abl, EVL, β-integrins.
Role: Project #3 Leader
1R21CA198472-01A1 (MPI: Cress and Knudsen) 04/01/2016 - 03/31/2018 1.2 CM
NIH/NCI $150,000/yr DC
Prostate Cancer Progression and Regulation of Laminin Adhesion”
Our previous work predicted that a translational control mechanism was responsible for the observed dysregulation of a major adhesion structure in prostate cancer. Recent genetic studies now show that specific microRNAs are aberrant in families with a congenital abnormality in this same adhesion structure. The hypothesis to be tested is that these specific microRNAs are responsible for the dysregulation found in prostate cancer.
2R01CA159406 (PI: Cress) 07/01/2016 - 06/30/2021 1.2 CM
NIH $250,000/yr DC
Human Prostate Cancer Metastasis and Laminin Binding Integrins
The goal of this renewal is to understand human prostate tumor cell adhesion, migration and use this information to block subsequent bone pain and spread to late stage secondary skeletal sites. The unique and innovative feature is pursuing the discovery of cohesive cell migration of micro metastases mediated by laminin binding integrins (LBI) and intercellular adhesion. Confocal live 3D imaging, translational pathology and state of the art mass spectrometry approaches will be used. The results will uncover new targetable elements of the LBI axis in prostate cancer progression.
See NIH BIO
See NIH BIO
Adv Topics in Cancer BiologyCBIO 553 (Spring 2018)
Directed ResearchBIOC 492 (Spring 2018)
DissertationCBIO 920 (Spring 2018)
ResearchCBIO 900 (Spring 2018)
Research ConferenceCBIO 695A (Spring 2018)
Cancer BiologyCBIO 552 (Fall 2017)
Directed ResearchBIOC 492 (Fall 2017)
Introduction to ResearchMCB 795A (Fall 2017)
ResearchCBIO 900 (Fall 2017)
Research ConferenceCBIO 695A (Fall 2017)
Adv Topics in Cancer BiologyCBIO 553 (Spring 2017)
DissertationCBIO 920 (Spring 2017)
ResearchCBIO 900 (Spring 2017)
Research ConferenceCBIO 695A (Spring 2017)
Cancer BiologyCBIO 552 (Fall 2016)
DissertationCBIO 920 (Fall 2016)
Introduction to ResearchMCB 795A (Fall 2016)
ResearchCBIO 900 (Fall 2016)
Research ConferenceCBIO 695A (Fall 2016)
- 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).More infoThe 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.
- Cress, A., Nagle, R. B., & Cress, A. E. (2011). Metastasis Update: Human Prostate Carcinoma Invasion via Tubulogenesis. Prostate cancer, 2011.More infoThis 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).More infoThe 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.
- 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).More infoMetastasizing 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.
- 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).More infoExpression 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.More infoExisting 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.
- 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).More infoOf 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).More infoLaminins 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.
- 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).More infoIntegrins 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.
- 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).More infoThe 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).More infoHuman 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).More infoCell 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.
- 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).More infoEpigenetic 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.
- 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).More infoDuring 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.