As a Developmental Biologist my primary interest over the years has been in identifying the mechanisms underlying the formation of the valves in the embryonic heart. As a graduate student I developed an in vitro assay to show that a process of epithelial-mesenchmal transition (EMT) in the valve forming regions of the heart was induced by a signal from the adjacent myocardium. As a faculty member, my lab first showed that the growth factor TGFbeta mediated this process. We went on to characterize the TGFbeta isoforms that did this as well as exploring the receptors and signal transduction pathways involved. Our most recent work has been on the role of a poorly characterized extracellular matrix protein, OLFM1, in reglating the invasion step of EMT. We have found the molecule to be present in several developmental EMTs and in cancer metastasis. Ongoing experiments are looking at this molecle in both the developing heart and in several cancers
A second project on the effects of trichloroethylene (TCE) on heart development began after my arrival in Arizona. It was observed that an increased number of heart defects were observed in an area of Tucson where the water supply had been contaminated with TCE. We set out to explore the cellular and molecular basis of this effect. We found that TCE alters calcium metabolism and myocardial contraction by altering transcription of many genes including molecules that mediate calcium release and uptake. Recent work on transcriptional reglation in the heart has also pointed to an effect on the transcription factor HNF4a.
A third project derives from the observation that stem cells and myocytes placed in proximity in culture show an improvement in myocyte viability and a unidirectional transfer of mitochondria from the stem cells to cardiomyocytes. We are curently exploring whether such transfer can take place in the intact heart after stem cell injection.
- Ph.D. Anatomy and Cell Biology
- Texas Tech University Health Sciences Center, Lubbock, Texas, United States
- Examination of a Tissue Interaction in Heart Development
- M.S. Biology (Genetics)
- Florida State University, Tallahassee, Florida, United States
- Ultrastructural Examination and Nutritional Regulation of Development in Allomyces arbuscula
- B.A. Biology
- Macalester College, St. Paul, Minnesota, United States
- University of Arizona, Tucson, Arizona (1992 - Ongoing)
- University of Iowa, Iowa City, Iowa (1986 - 1992)
- R.R. Bensley award
- American Association of Anatomists, Spring 1992
- Established Investigator
- American Heart Association, Spring 1991
- Markwald Award
- Weinstein Cardiovascular Development and Regeneration Conference, Spring 2019
- American Academy of Regenerative Medicine, Spring 2016
- Alumni election to Phi Beta Kappa
- Macalester College, Epsilon of Minnesota, Spring 2014
- Alumni Service Award
- Macalester College, Summer 2010
- Fellow of the American Association of Anatomists
- American Association of Anatomy, Spring 2010
- The Elsie Taber Lectureship in Human Development
- Medical University of South Carolina, Spring 2007
- Distinguished Alumni Award of the Graduate School of Biomedical Sciences
- Texas Tech University Health Sciences Center, Fall 2003
- 2003 Distinguished Alumni Award, Graduate School of Biomedical Sciences
- Texas Tech University, Spring 2003
No activities entered.
Directed ResearchNSCS 492 (Fall 2021)
Independent StudyNSCS 499 (Fall 2021)
Cell Biology of DiseaseCMM 404 (Summer I 2021)
Cell Biology of DiseaseCMM 504 (Summer I 2021)
EMT in Development and DiseaseCMM 558 (Spring 2021)
ThesisCMM 910 (Spring 2021)
Working Clinical CasesCMM 461 (Spring 2021)
Working Clinical CasesCMM 561 (Spring 2021)
Cell Biology of DiseaseCMM 504 (Summer I 2020)
Cardiovascular BiologyCMM 596A (Spring 2020)
Cardiovascular BiologyCMM 596A (Fall 2019)
EMT in Development and DiseaseCMM 558 (Fall 2019)
Cell Biology of DiseaseCMM 404 (Summer I 2019)
Cell Biology of DiseaseCMM 504 (Summer I 2019)
Honors ThesisECOL 498H (Spring 2019)
Rsrch Meth Psio SciPS 700 (Spring 2019)
Smnr Cardiovascular DevCMM 596A (Spring 2019)
Honors ThesisECOL 498H (Fall 2018)
Smnr Cardiovascular DevCMM 596A (Fall 2018)
Cell Biology of DiseaseCMM 504 (Summer I 2018)
Honors Independent StudyECOL 399H (Summer I 2018)
Honors Independent StudyECOL 399H (Spring 2018)
Smnr Cardiovascular DevCMM 596A (Spring 2018)
Honors Independent StudyECOL 399H (Fall 2017)
Smnr Cardiovascular DevCMM 596A (Fall 2017)
Cell Biology of DiseaseCMM 404 (Summer I 2017)
Cell Biology of DiseaseCMM 504 (Summer I 2017)
Honors Independent StudyNSCS 299H (Summer I 2017)
Directed RsrchMCB 492 (Spring 2017)
Honors Independent StudyECOL 399H (Spring 2017)
Honors ThesisMCB 498H (Spring 2017)
Smnr Cardiovascular DevCMM 596A (Spring 2017)
Honors ThesisMCB 498H (Fall 2016)
Smnr Cardiovascular DevCMM 596A (Fall 2016)
Cell Biology of DiseaseCMM 404 (Summer I 2016)
Cell Biology of DiseaseCMM 504 (Summer I 2016)
Honors Independent StudyECOL 399H (Spring 2016)
Honors ThesisMCB 498H (Spring 2016)
Senior CapstoneMCB 498 (Spring 2016)
Smnr Cardiovascular DevCMM 596A (Spring 2016)
- Selmin, O. I., Runyan, R. B., & Makwana, O. (2014). Environmental Sensitivity to Trichloroethylene (TCE) in the Developing Heart. In Trichloroethylene: Toxicity and Health Risks(pp 153-169). London: Springer.
- Nfonsam, V. N., Nfonsam, L. E., Chen, D., Omesiete, P. N., Cruz, A., Runyan, R. B., & Jandova, J. (2019). COMP Gene Coexpresses With EMT Genes and Is Associated With Poor Survival in Colon Cancer Patients. JOURNAL OF SURGICAL RESEARCH, 233, 297-303.
- Runyan, R. B., Selmin, O. I., Smith, S. M., & Freeman, J. L. (2019). Letter to the Editor. BIRTH DEFECTS RESEARCH, 111(16), 1234-1236.More infoThis letter was written in response to a publication produced by the Halogenated Solvents Industry Alliance to support of rolling back EPA standards for exposure to trichloroethylene. The letter emphasized the body of data produced in our labs and others that showed the solvent produces gene transcription and functional effects on heart development at doses significantly lower than tested in their publication.
- Yuan, C., Wang, Z., Borg, T. K., Ye, T., Baicu, C., Bradshaw, A., Zile, M., Runyan, R. B., Shao, Y., & Ga0, B. Z. (2019). Changes in the crystallographic structures of cardiac myosin filaments detected by polarization-dependent second harmonic generation microscopy. Biomedical Optics Express, 10(7), 3183-3195. doi:•https://doi.org/10.1364/BOE.10.003183
- Harris, A. P., Ismail, K. A., Nunez, M., Martopullo, I., Lencinas, A., Selmin, O. I., & Runyan, R. B. (2018). Trichloroethylene perturbs HNF4a expression and activity in the developing chick heart. Toxicology letters, 285, 113-120.More infoExposure to trichloroethylene (TCE) is linked to formation of congenital heart defects in humans and animals. Prior interactome analysis identified the transcription factor, Hepatocyte Nuclear Factor 4 alpha (HNF4a), as a potential target of TCE exposure. As a role for HNF4a is unknown in the heart, we examined developing avian hearts for HNF4a expression and for sensitivity to TCE and the HNF4a agonist, Benfluorex. In vitro analysis using a HNF4a reporter construct showed both TCE and HFN4a to be antagonists of HNF4a-mediated transcription at the concentrations tested. HNF4a mRNA is expressed transiently in the embryonic heart during valve formation and cardiac development. Embryos were examined for altered gene expression in the presence of TCE or Benfluorex. TCE altered expression of selected mRNAs including HNF4a, TRAF6 and CYP2C45. There was a transition between inhibition and induction of marker gene expression in embryos as TCE concentration increased. Benfluorex was largely inhibitory to selected markers. Echocardiography of exposed embryos showed reduced cardiac function with both TCE and Benfluorex. Cardiac contraction was reduced by 29% and 23%, respectively at 10 ppb. The effects of TCE and Benfluorex on autocrine regulation of HNF4a, selected markers and cardiac function argue for a functional interaction of TCE and HNF4a. Further, the dose-sensitive shift between inhibition and induction of marker expression may explain the nonmonotonic-like dose response observed with TCE exposure in the heart.
- Runyan, R. B. (2018). Remodeling Failing Human Myocardium With Hybrid Cell/Matrix and Transmyocardial Revascularization. ASAIO JOURNAL, 64(5), E130-E133. doi:10.1097/MAT.0000000000000727More infoGiven the limited treatment options for advanced heart failure, the intrinsic regenerative properties of stem cells have been evaluated for myocardial remodeling. Previous stem cells techniques for myocardiocyte remodeling have been limited by the low cellular retention. Presented is a hybrid approach for remodeling infarcted myocardium through implantation of allogeneic human amniotic fluid–derived mesenchymal stem cells within micronized human allograft-derived liquid matrix during the performance of transmyocardial revascularization (TMR). Given the induced increase in vascular density from TMR, we hypothesize that it may serve as a therapeutic delivery system for stem cell placement into damaged myocardium. We present a patient with ischemic cardiomyopathy and refractory angina, who clinically improved after this hybrid therapy of intraoperative TMR and placement of amniotic fluid–derived mesenchymal stem cells and liquid matrix within the TMR channels. Noninvasive testing of myocardial viability biomarkers utilizing both cardiac magnetic resonance imaging and thallium imaging supported the clinical improvement in cardiac symptom may be related to ventricular remodeling in a region of infarct with subsequent functional improvement.
- Runyan, R. B., & Savagner, P. (2018). Epithelial-mesenchymal transition and plasticity in the developmental basis of cancer and fibrosis. DEVELOPMENTAL DYNAMICS, 247(3), 330-331.
- Runyan, R. B., & Savagner, P. (2018). Epithelial-mesenchymal transition and plasticity in the developmental basis of cancer and fibrosis. Developmental dynamics : an official publication of the American Association of Anatomists, 247(3), 330-331.
- Tavares, A. L., Brown, J. A., Ulrich, E. C., Dvorak, K., & Runyan, R. B. (2018). Runx2-I is an Early Regulator of Epithelial-Mesenchymal Cell Transition in the Chick Embryo. Developmental dynamics : an official publication of the American Association of Anatomists, 247(3), 542-554.More infoAlthough normally linked to bone and cartilage development, the Runt-related transcription factor, RUNX2, was reported in the mouse heart during development of the valves. We examined RUNX2 expression and function in the developing avian heart as it related to the epithelial-mesenchymal transition (EMT) in the atrioventricular canal. EMT can be separated into an activation stage involving hypertrophy and cell separation and an invasion stage where cells invade the extracellular matrix. The localization and activity of RUNX2 was explored in relation to these steps in the heart. As RUNX2 was also reported in cancer tissues, we examined its expression in the progression of esophageal cancer in staged tissues.
- Tavares, A., Brown, J. A., Ulrich, E. C., Dvorak, K., & Runyan, R. B. (2018). Runx2-I is an Early Regulator of Epithelial-Mesenchymal Cell Transition in the Chick Embryo. DEVELOPMENTAL DYNAMICS, 247(3), 542-554.
- Ferng, A., Lick, S. D., Desai, A., Kazui, T., Runyan, R. B., & Khalpey, Z. I. (2017). Cardiac regeneration in the human left ventricle after CorMatrix implantation. Ann Thorac Surg, 104(3), e239-e241.
- Iwanski, J., Iwanski, J., Knapp, S. M., Knapp, S. M., Avery, R., Avery, R., Oliva, I. B., Oliva, I. B., Wong, R. K., Wong, R. K., Runyan, R. B., Runyan, R. B., Khalpey, Z. I., & Khalpey, Z. I. (2017). Clinical outcomes meta-analysis: measuring subendocardial perfusion and efficacy of transmyocardial laser revascularization with nuclear imaging. Journal of Cardiothoracic Surgery, 12(1). doi:10.1186/s13019-017-0602-8
- Iwanski, J., Knapp, S. M., Avery, R., Oliva, I., Wong, R. K., Runyan, R. B., & Khalpey, Z. (2017). Clinical outcomes meta-analysis: measuring subendocardial perfusion and efficacy of transmyocardial laser revascularization with nuclear imaging. JOURNAL OF CARDIOTHORACIC SURGERY, 12.
- Kazui, T., Tran, P. L., Pilikian, T. R., Marsh, K. M., Runyan, R., Konhilas, J., Smith, R., & Khalpey, Z. I. (2017). A dual therapy of off-pump temporary left ventricular extracorporeal device and amniotic stem cell for cardiogenic shock. Journal of cardiothoracic surgery, 12(1), 80.More infoTemporary mechanical circulatory support device without sternotomy has been highly advocated for severe cardiogenic shock patient but little is known when coupled with amniotic stem cell therapy.
- Schipper, D. A., Louis, A. V., Dicken, D. S., Johnson, K., Smolenski, R. T., Black, S. M., Runyan, R., Konhilas, J., Garcia, J. G., & Khalpey, Z. (2017). Improved metabolism and redox state with a novel preservation solution: implications for donor lungs after cardiac death (DCD). Pulmonary circulation, 7(2), 494-504.More infoLungs donated after cardiac death (DCD) are an underutilized resource for a dwindling donor lung transplant pool. Our study investigates the potential of a novel preservation solution, Somah, to better preserve statically stored DCD lungs, for an extended time period, when compared to low-potassium dextran solution (LPD). We hypothesize that Somah is a metabolically superior organ preservation solution for hypothermic statically stored porcine DCD lungs, possibly improving lung transplant outcomes. Porcine DCD lungs (n = 3 per group) were flushed with and submerged in cold preservation solution. The lungs were stored up to 12 h, and samples were taken from lung tissue and the preservation medium throughout. Metabolomic and redox potential were analyzed using high performance liquid chromatography, mass spectrometry, and RedoxSYS®, comparing substrate and pathway utilization in both preservation solutions. Glutathione reduction was seen in Somah but not in LPD during preservation. Carnitine, carnosine, and n-acetylcarnosine levels were elevated in the Somah medium compared with LPD throughout. Biopsies of Somah exposed lungs demonstrated similar trends after 2 h, up to 12 h. Adenosine gradually decreased in Somah medium over 12 h, but not in LPD. An inversely proportional increase in inosine was found in Somah. Higher oxidative stress levels were measured in LPD. Our study suggests suboptimal metabolic preservation in lungs stored in LPD. LPD had poor antioxidant potential, cytoprotection, and an insufficient redox potential. These findings may have immediate clinical implications for human organs; however, further investigation is needed to evaluate DCD lung preservation in Somah as a viable option for transplant.
- Ferng, A. S., Marsh, K. M., Fleming, J. M., Schipper, D., Bajaj, N., Connell, A. M., Pilikan, T., Johnson, K., Runyan, R. B., Black, S. M., Szivek, J. A., & Khalpey, Z. I. (2016). Adipose-Derived Human Stem Cells: Comparative Organ Specific Mitochondrial Bioenergy Profiles. SpringerPlus, 5(2057). doi:10.1186/s40064-016-3712-1
- Iwanski, J., Wong, R. K., Larson, D. F., Ferng, A. S., Runyan, R. B., Goldstein, S., & Khalpey, Z. (2016). Remodeling an infarcted heart: novel hybrid treatment with transmyocardial revascularization and stem cell therapy. SPRINGERPLUS, 5.
- Yang, H., Borg, T. K., Ma, Z., Xu, M., Wetzel, G., Saraf, L. V., Markwald, R., Runyan, R. B., & Gao, B. Z. (2016). Biochip-based study of unidirectional mitochondrial transfer from stem cells to myocytes via tunneling nanotubes. Biofabrication, 8(1), 015012.More infoTunneling nanotubes (TNTs) are small membranous tubes of 50-1000 nm diameter observed to connect cells in culture. Transfer of subcellular organelles through TNTs was observed in vitro and in vivo, but the formation and significance of these structures is not well understood. A polydimethylsiloxane biochip-based coculture model was devised to constrain TNT orientation and explore both TNT-formation and TNT-mediated mitochondrial transfer. Two parallel microfluidic channels connected by an array of smaller microchannels enabled localization of stem cell and cardiomyocyte populations while allowing connections to form between them. Stem cells and cardiomyocytes were deposited in their respective microfluidic channels, and stem cell-cardiomyocyte pairs were formed via the microchannels. Formation of TNTs and transfer of stained mitochondria through TNTs was observed by 24 h real-time video recording. The data show that stem cells are 7.7 times more likely to initiate contact by initial extension of filopodia. By 24 h, 67% of nanotube connections through the microchannels are composed of cardiomyocyte membrane. Filopodial extension and retraction by stem cells draws an extension of TNTs from cardiomyocytes. MitoTracker staining shows that unidirectional transfer of mitochondria between stem cell-cardiomyocyte pairs invariably originates from stem cells. Control experiments with cardiac fibroblasts and cardiomyocytes show little nanotube formation between homotypic or mixed cell pairs and no mitochondrial transfer. These data identify a novel biological process, unidirectional mitochondrial transfer, mediated by heterotypic TNT connections. This suggests that the enhancement of cardiomyocyte function seen after stem-cell injection may be due to a bioenergetic stimulus provided by mitochondrial transfer.
- Yang, H., Schmidt, L. P., Wang, Z., Yang, X., Shao, Y., Borg, T. K., Markwald, R., Runyan, R., & Gao, B. Z. (2016). Dynamic Myofibrillar Remodeling in Live Cardiomyocytes under Static Stretch. Scientific reports, 6, 20674.More infoAn increase in mechanical load in the heart causes cardiac hypertrophy, either physiologically (heart development, exercise and pregnancy) or pathologically (high blood pressure and heart-valve regurgitation). Understanding cardiac hypertrophy is critical to comprehending the mechanisms of heart development and treatment of heart disease. However, the major molecular event that occurs during physiological or pathological hypertrophy is the dynamic process of sarcomeric addition, and it has not been observed. In this study, a custom-built second harmonic generation (SHG) confocal microscope was used to study dynamic sarcomeric addition in single neonatal CMs in a 3D culture system under acute, uniaxial, static, sustained stretch. Here we report, for the first time, live-cell observations of various modes of dynamic sarcomeric addition (and how these real-time images compare to static images from hypertrophic hearts reported in the literature): 1) Insertion in the mid-region or addition at the end of a myofibril; 2) Sequential addition with an existing myofibril as a template; and 3) Longitudinal splitting of an existing myofibril. The 3D cell culture system developed on a deformable substrate affixed to a stretcher and the SHG live-cell imaging technique are unique tools for real-time analysis of cultured models of hypertrophy.
- Cox, C. M., Mandell, E. K., Stewart, L., Lu, R., Johnson, D. L., McCarter, S. D., Tavares, A., Runyan, R., Ghosh, S., & Wilson, J. M. (2015). Endosomal regulation of contact inhibition through the AMOT:YAP pathway. Molecular biology of the cell, 26(14), 2673-84.More infoContact-mediated inhibition of cell proliferation is an essential part of organ growth control; the transcription coactivator Yes-associated protein (YAP) plays a pivotal role in this process. In addition to phosphorylation-dependent regulation of YAP, the integral membrane protein angiomotin (AMOT) and AMOT family members control YAP through direct binding. Here we report that regulation of YAP activity occurs at the endosomal membrane through a dynamic interaction of AMOT with an endosomal integral membrane protein, endotubin (EDTB). EDTB interacts with both AMOT and occludin and preferentially associates with occludin in confluent cells but with AMOT family members in subconfluent cells. EDTB competes with YAP for binding to AMOT proteins in subconfluent cells. Overexpression of the cytoplasmic domain or full-length EDTB induces translocation of YAP to the nucleus, an overgrowth phenotype, and growth in soft agar. This increase in proliferation is dependent upon YAP activity and is complemented by overexpression of p130-AMOT. Furthermore, overexpression of EDTB inhibits the AMOT:YAP interaction. EDTB and AMOT have a greater association in subconfluent cells compared with confluent cells, and this association is regulated at the endosomal membrane. These data provide a link between the trafficking of tight junction proteins through endosomes and contact-inhibition-regulated cell growth.
- Caldwell, P. T., Manziello, A., Howard, J., Palbykin, B., Runyan, R. B., & Selmin, O. (2013). Gene Expression Profiling in the Fetal Cardiac Tissue after Folate and Low-Dose Trichloroethylene Exposure. BIRTH DEFECTS RESEARCH PART A-CLINICAL AND MOLECULAR TERATOLOGY, 88(2), 111-127.
- Lencinas, A., Broka, D. M., Konieczka, J. H., Klewer, S. E., Antin, P. B., Camenisch, T. D., & Runyan, R. B. (2013). Arsenic Exposure Perturbs Epithelial-Mesenchymal Cell Transition and Gene Expression In a Collagen Gel Assay. TOXICOLOGICAL SCIENCES, 116(1), 273-285.
- Liu, H., Shao, Y., Qin, W., Runyan, R. B., Xu, M., Ma, Z., Borg, T. K., Markwald, R., & Gao, B. Z. (2013). Myosin filament assembly onto myofibrils in live neonatal cardiomyocytes observed by TPEF-SHG microscopy. Cardiovascular research, 97(2), 262-70.More infoUnderstanding myofibrillogenesis is essential for elucidating heart muscle formation, development, and remodelling in response to physiological stimulation. Here, we report the dynamic assembly process of contractile myosin filaments onto myofibrils in a live cardiomyocyte culture during myofibrillogenesis.
- Ma, Z., Liu, Q., Yang, H., Runyan, R. B., Eisenberg, C. A., Xu, M., Borg, T. K., Markwald, R., Wang, Y., & Gao, B. Z. (2013). Laser patterning for the study of MSC cardiogenic differentiation at the single-cell level. Light, science & applications, 2.More infoMesenchymal stem cells (MSCs) have been cited as contributors to heart repair through cardiogenic differentiation and multiple cellular interactions, including the paracrine effect, cell fusion, and mechanical and electrical couplings. Due to heart-muscle complexity, progress in the development of knowledge concerning the role of MSCs in cardiac repair is heavily based on MSC-cardiomyocyte coculture. In conventional coculture systems, however, the in vivo cardiac muscle structure, in which rod-shaped cells are connected end-to-end, is not sustained; instead, irregularly shaped cells spread randomly, resulting in randomly distributed cell junctions. Consequently, contact-mediated cell-cell interactions (e.g., the electrical triggering signal and the mechanical contraction wave that propagate through MSC-cardiomyocyte junctions) occur randomly. Thus, the data generated on the beneficial effects of MSCs may be irrelevant to in vivo biological processes. In this study, we explored whether cardiomyocyte alignment, the most important phenotype, is relevant to stem cell cardiogenic differentiation. Here, we report (i) the construction of a laser-patterned, biochip-based, stem cell-cardiomyocyte coculture model with controlled cell alignment; and (ii) single-cell-level data on stem cell cardiogenic differentiation under in vivo-like cardiomyocyte alignment conditions.
- Ma, Z., Yang, H., Liu, H., Xu, M., Runyan, R. B., Eisenberg, C. A., Markwald, R. R., Borg, T. K., & Gao, B. Z. (2013). Mesenchymal stem cell-cardiomyocyte interactions under defined contact modes on laser-patterned biochips. PloS one, 8(2), e56554.More infoUnderstanding how stem cells interact with cardiomyocytes is crucial for cell-based therapies to restore the cardiomyocyte loss that occurs during myocardial infarction and other cardiac diseases. It has been thought that functional myocardial repair and regeneration could be regulated by stem cell-cardiomyocyte contact. However, because various contact modes (junction formation, cell fusion, partial cell fusion, and tunneling nanotube formation) occur randomly in a conventional coculture system, the particular regulation corresponding to a specific contact mode could not be analyzed. In this study, we used laser-patterned biochips to define cell-cell contact modes for systematic study of contact-mediated cellular interactions at the single-cell level. The results showed that the biochip design allows defined stem cell-cardiomyocyte contact-mode formation, which can be used to determine specific cellular interactions, including electrical coupling, mechanical coupling, and mitochondria transfer. The biochips will help us gain knowledge of contact-mediated interactions between stem cells and cardiomyocytes, which are fundamental for formulating a strategy to achieve stem cell-based cardiac tissue regeneration.
- Runyan, R., Lencinas, A., Chhun, D. C., Dan, K. P., Ross, K. D., Hoover, E. A., Antin, P. B., & Runyan, R. B. (2013). Olfactomedin-1 activity identifies a cell invasion checkpoint during epithelial-mesenchymal transition in the chick embryonic heart. Disease models & mechanisms, 6(3).More infoEndothelia in the atrioventricular (AV) canal of the developing heart undergo a prototypical epithelial mesenchymal transition (EMT) to begin heart valve formation. Using an in vitro invasion assay, an extracellular matrix protein, Olfactomedin-1 (OLFM1), was found to increase mesenchymal cell numbers in AV canals from embryonic chick hearts. Treatment with both anti-OLFM1 antibody and siRNA targeting OLFM1 inhibits mesenchymal cell formation. OLFM1 does not alter cell proliferation, migration or apoptosis. Dispersion, but lack of invasion in the presence of inhibiting antibody, identifies a specific role for OLFM1 in cell invasion during EMT. This role is conserved in other epithelia, as OLFM1 similarly enhances invasion by MDCK epithelial cells in a transwell assay. Synergy is observed when TGFβ2 and OLFM1 are added to MDCK cell cultures, indicating that OLFM-1 activity is cooperative with TGFβ. Inhibition of both OLFM1 and TGFβ in heart invasion assays shows a similar cooperative role during development. To explore OLFM1 activity during EMT, representative EMT markers were examined. Effects of OLFM1 protein and anti-OLFM1 on transcripts of cell-cell adhesion molecules and the transcription factors Snail-1, Snail-2, Twist1 and Sox-9 argue that OLFM1 does not initiate EMT. Rather, regulation of transcripts of Zeb1 and Zeb2, secreted proteases and mesenchymal cell markers by both OLFM1 and anti-OLFM1 is consistent with regulation of the cell invasion step of EMT. We conclude that OLFM1 is present and necessary during EMT in the embryonic chick heart. Its role in cell invasion and mesenchymal cell gene regulation suggests an invasion checkpoint in EMT where OLFM1 acts to promote cell invasion into the three-dimensional matrix.
- Runyan, R., Makwana, O., Ahles, L., Lencinas, A., Selmin, O. I., & Runyan, R. B. (2013). Low-dose trichloroethylene alters cytochrome P450-2C subfamily expression in the developing chick heart. Cardiovascular toxicology, 13(1).More infoTrichloroethylene (TCE) is an organic solvent and common environmental contaminant. TCE exposure is associated with heart defects in humans and animal models. Primary metabolism of TCE in adult rodent models is by specific hepatic cytochrome P450 enzymes (Lash et al. in Environ Health Perspect 108:177-200, 2000). As association of TCE exposure with cardiac defects is in exposed embryos prior to normal liver development, we investigated metabolism of TCE in the early embryo. Developing chick embryos were dosed in ovo with environmentally relevant doses of TCE (8 and 800 ppb) and RNA was extracted from cardiac and extra-cardiac tissue (whole embryo without heart). Real-time PCR showed upregulation of CYP2H1 transcripts in response to TCE exposure in the heart. No detectable cytochrome expression was found in extra-cardiac tissue. As seen previously, the dose response was non-monotonic and 8 ppb elicited stronger upregulation than 800 ppb. Immunostaining for CYP2C subfamily expression confirmed protein expression and showed localization in both myocardium and endothelium. TCE exposure increased protein expression in both tissues. These data demonstrate that the earliest embryonic expression of phase I detoxification enzymes is in the developing heart. Expression of these CYPs is likely to be relevant to the susceptibility of the developing heart to environmental teratogens.
- Doyle, S. E., Scholz, M. J., Greer, K. A., Hubbard, A. D., Darnell, D. K., Antin, P. B., Klewer, S. E., & Runyan, R. B. (2011). Latrophilin-2 is a novel component of the epithelial-mesenchymal transition within the atrioventricular canal of the embryonic chicken heart. DEVELOPMENTAL DYNAMICS, 235(12), 3213-3221.
- Ma, Z., Yang, H., Liu, H., Xu, M., Runyan, R. B., Eisenberg, C. A., Markwald, R. R., Borg, T. K., & Gao, B. Z. (2011). Mesenchymal Stem Cell-Cardiomyocyte Interactions under Defined Contact Modes on Laser-Patterned Biochips. PLOS ONE, 8(2).
- Runyan, R., Lencinas, A., Tavares, A. L., Barnett, J. V., & Runyan, R. B. (2011). Collagen gel analysis of epithelial-mesenchymal transition in the embryo heart: an in vitro model system for the analysis of tissue interaction, signal transduction, and environmental effects. Birth defects research. Part C, Embryo today : reviews, 93(4).More infoThe cellular process of epithelial-mesenchymal cell transition (EMT) is a critical event in development that is reiterated in adult pathologies of metastasis and organ fibrosis. An initial understanding of the cellular and molecular events of this process emerged from an in vitro examination of heart valve development. Explants of the chick atrioventricular valve-forming region were placed on collagen gels and removed to show that EMT was regulated by a tissue interaction. Subsequent studies showed that specific TGFβ isoforms and receptors were required and steps of activation and invasion could be distinguished. The assay was modified for mouse hearts and has been used to explore signal transduction and gene expression in both species. The principle advantages of the system are a defined temporal window, when EMT takes place and the ability to isolate cells at various stages of the EMT process. These advantages are largely unavailable in other developmental or adult models. As the mesenchymal cells produced by EMT in the heart are involved in defects found in congenital heart disease, there is also a direct relevance of cardiac EMT to human birth defects. This relationship has been explored in relation to environmental exposures and in a number of genetic models. This review provides both an overview of the findings developed from the assay and protocols to enable the use of the assay by other laboratories. The assay provides a versatile platform to explore roles of specific gene products, drugs, and environmental agents on a critical cellular process.
- Caldwell, P. T., Manziello, A., Howard, J., Palbykin, B., Runyan, R. B., & Selmin, O. (2010). Gene expression profiling in the fetal cardiac tissue after folate and low-dose trichloroethylene exposure. Birth defects research. Part A, Clinical and molecular teratology, 88(2), 111-27.More infoPrevious studies show gene expression alterations in rat embryo hearts and cell lines that correspond to the cardio-teratogenic effects of trichloroethylene (TCE) in animal models. One potential mechanism of TCE teratogenicity may be through altered regulation of calcium homeostatic genes with a corresponding inhibition of cardiac function. It has been suggested that TCE may interfere with the folic acid/methylation pathway in liver and kidney and alter gene regulation by epigenetic mechanisms. According to this hypothesis, folate supplementation in the maternal diet should counteract TCE effects on gene expression in the embryonic heart.
- Mercado-Pimentel, M. E., Hubbard, A. D., & Runyan, R. B. (2010). Endoglin and Alk5 regulate epithelial-mesenchymal transformation during cardiac valve formation. DEVELOPMENTAL BIOLOGY, 304(1), 420-432.
- Runyan, R., Lencinas, A., Broka, D. M., Konieczka, J. H., Klewer, S. E., Antin, P. B., Camenisch, T. D., & Runyan, R. B. (2010). Arsenic exposure perturbs epithelial-mesenchymal cell transition and gene expression in a collagen gel assay. Toxicological sciences : an official journal of the Society of Toxicology, 116(1).More infoArsenic is a naturally occurring metalloid and environmental contaminant. Arsenic exposure in drinking water is reported to cause cancer of the liver, kidneys, lung, bladder, and skin as well as birth defects, including neural tube, facial, and vasculogenic defects. The early embryonic period most sensitive to arsenic includes a variety of cellular processes. One key cellular process is epithelial-mesenchymal transition (EMT) where epithelial sheets develop into three-dimensional structures. An embryonic prototype of EMT is found in the atrioventricular (AV) canal of the developing heart, where endothelia differentiate to form heart valves. Effects of arsenic on this cellular process were examined by collagen gel invasion assay (EMT assay) using explanted AV canals from chicken embryo hearts. AV canals treated with 12.5-500 ppb arsenic showed a loss of mesenchyme at 12.5 ppb, and mesenchyme formation was completely inhibited at 500 ppb. Altered gene expression in arsenic-treated explants was investigated by microarray analysis. Genes whose expression was altered consistently at exposure levels of 10, 25, and 100 ppb were identified, and results showed that 25 ppb in vitro was particularly effective. Three hundred and eighty two genes were significantly altered at this exposure level. Cytoscape analysis of the microarray data using the chicken interactome identified four clusters of altered genes based on published relationships and pathways. This analysis identified cytoskeleton and cell adhesion-related genes whose disruption is consistent with an altered ability to undergo EMT. These studies show that EMT is sensitive to arsenic and that an interactome-based approach can be useful in identifying targets.
- Runyan, R., Makwana, O., King, N. M., Ahles, L., Selmin, O., Granzier, H. L., & Runyan, R. B. (2010). Exposure to low-dose trichloroethylene alters shear stress gene expression and function in the developing chick heart. Cardiovascular toxicology, 10(2).More infoTrichloroethylene is an organic solvent used as an industrial degreasing agent. Due to its widespread use and volatile nature, TCE is a common environmental contaminant. Trichloroethylene exposure has been implicated in the etiology of heart defects in human populations and animal models. Recent data suggest misregulation of Ca2+ homeostasis in H9c2 cardiomyocyte cell line after TCE exposure. We hypothesized that misregulation of Ca2+ homeostasis alters myocyte function and leads to changes in embryonic blood flow. In turn, changes in cardiac flow are known to cause cardiac malformations. To investigate this hypothesis, we dosed developing chick embryos in ovo with environmentally relevant doses of TCE (8 and 800 ppb). RNA was isolated from control and treated embryos at specific times in development for real-time PCR analysis of blood flow markers. Effects were observed on Endothelin-1 (ET-1), Nitric Oxide Synthase-3 (NOS-3) and Krüppel-like Factor 2 (KLF2) expression relative to TCE exposure and consistent with reduced flow. Further, we measured function in the developing heart after TCE exposure by isolating cardiomyocytes and measuring half-width of contraction and sarcomere lengths. These functional data showed a significant increase in half-width of contraction after TCE exposure. These data suggest that perturbation of cardiac function contributes to the etiology of congenital heart defects in TCE-exposed embryos.
- Azhar, M., Runyan, R. B., Gard, C., Sanford, L. P., Miller, M. L., Andringa, A., Pawlowski, S., Rajan, S., & Doetschman, T. (2009). Ligand-specific function of transforming growth factor beta in epithelial-mesenchymal transition in heart development. Developmental dynamics : an official publication of the American Association of Anatomists, 238(2), 431-42.More infoThe ligand specificity of transforming growth factor beta (TGFbeta) in vivo in mouse cardiac cushion epithelial-to-mesenchymal transition (EMT) is poorly understood. To elucidate the function of TGFbeta in cushion EMT, we analyzed Tgfb1(-/-), Tgfb2(-/-), and Tgfb3(-/-) mice between embryonic day (E) 9.5 and E14.5 using both in vitro and in vivo approaches. Atrioventricular (AV) canal collagen gel assays at E9.5 indicated normal EMT in both Tgfb1(-/-) and Tgfb3(-/-) mice. However, analysis of Tgfb2(-/-) AV explants at E9.5 and E10.5 indicated that EMT, but not cushion cell proliferation, was initially delayed but later remained persistent. This was concordant with the observation that Tgfb2(-/-) embryos, and not Tgfb1(-/-) or Tgfb3(-/-) embryos, develop enlarged cushions at E14.5 with elevated levels of well-validated indicators of EMT. Collectively, these data indicate that TGFbeta2, and not TGFbeta1 or TGFbeta3, mediates cardiac cushion EMT by promoting both the initiation and cessation of EMT.
- Ma, Z., Liu, Q., Yang, H., Runyan, R. B., Eisenberg, C. A., Xu, M., Borg, T. K., Markwald, R., Wang, Y., & Gao, B. Z. (2009). Laser patterning for the study of MSC cardiogenic differentiation at the single-cell level. LIGHT-SCIENCE & APPLICATIONS, 2.
- O'Connell, D. J., Molinar, A. J., Tavares, A. L., Mathine, D. L., Runyan, R. B., & Bahl, J. J. (2009). Transfection of cells attached to selected cell based biosensor surfaces. LIFE SCIENCES, 80(15), 1395-1402.
- Rausch, M. P., Hahn, T., Ramanathapuram, L., Bradley-Dunlop, D., Mahadevan, D., Mercado-Pimentel, M. E., Runyan, R. B., Besselsen, D. G., Zhang, X., Cheung, H. K., Lee, W. C., Ling, L. E., & Akporiaye, E. T. (2009). An orally active small molecule TGF-beta receptor I antagonist inhibits the growth of metastatic murine breast cancer. Anticancer research, 29(6), 2099-109.More infoTransforming growth factor beta (TGF-beta) plays a complex role in breast carcinogenesis. Initially functioning as a tumor suppressor, this cytokine later contributes to the progression of malignant cells by enhancing their invasive and metastatic potential as well as suppressing antitumor immunity. The purpose of this study was to investigate the efficacy of SM16, a novel small molecule ALK5 kinase inhibitor, to treat a highly metastatic, TGF-beta-producing murine mammary carcinoma (4T1).
- Caldwell, P. T., Thorne, P. A., Johnson, P. D., Boitano, S., Runyan, R. B., & Selmin, O. (2008). Trichloroethylene disrupts cardiac gene expression and calcium homeostasis in rat myocytes. Toxicological sciences : an official journal of the Society of Toxicology, 104(1), 135-43.More infoWe have been investigating the molecular mechanisms by which trichloroethylene (TCE) might induce cardiac malformations in the embryonic heart. Previous results indicated that TCE disrupted expression of genes encoding proteins involved in regulation of intracellular Ca2+, [Ca2+](i), in cardiac cells, including ryanodine receptor isoform 2 (Ryr2), and sarcoendoplasmatic reticulum Ca2+ ATPase, Serca2a. These observations are important in light of the notion that altered cardiac contractility can produce morphological defects. The hypothesis tested in this study is that the TCE-induced changes in gene expression of Ca2+-associated proteins resulted in altered Ca2+ flux regulation. We used real-time PCR and digital imaging microscopy to characterize effects of various doses of TCE on gene expression and Ca2+ response to vasopressin (VP) in rat cardiac H9c2 myocytes. We observed a reduction in Serca2a and Ryr2 expression at 12 and 48 h after exposure to TCE. In addition, we found significant differences in Ca2+ response to VP in cells treated with TCE doses as low as 10 parts per billion. Taken all together, our data strongly indicate that exposure to TCE disrupts the ability of myocytes to regulate cellular Ca2+ fluxes. Perturbation of calcium signaling alters cardiac cell physiology and signal transduction and may hint to morphogenetic consequences in the context of heart development. These results point to a novel area of TCE biology and, if confirmed in vivo, may help to explain the apparent cardio-specific toxicity of TCE exposure in the rodent embryo.
- Azhar, M., Runyan, R. B., Gard, C., Sanford, L. P., Miller, M. L., Andringa, A., Pawlowski, S., Rajan, S., & Doetschman, T. (2007). Ligand-Specific Function of Transforming Growth Factor Beta in Epithelial-Mesenchymal Transition in Heart Development. DEVELOPMENTAL DYNAMICS, 238(2), 431-442.
- Mercado-Pimentel, M. E., & Runyan, R. B. (2007). Multiple transforming growth factor-beta isoforms and receptors function during epithelial-mesenchymal cell transformation in the embryonic heart. CELLS TISSUES ORGANS, 185(1-3), 146-156.
- O'Connell, D. J., Molinar, A. J., Tavares, A. L., Mathine, D. L., Runyan, R. B., & Bahl, J. J. (2007). Transfection of cells attached to selected cell based biosensor surfaces. Life sciences, 80(15), 1395-402.More infoMammalian cell attachment studies were conducted on a variety of common microchip surfaces for potential use in cell based biosensors. COS-7 cell attachment to Au, Pt or ITO, per unit area was greater than to SiO(2) surfaces. The number of cells that would attach was essentially maximized 3 h after cell seeding. HL-1 cells attached more readily to surfaces precoated with fibronectin, but by 3 h equivalent number of cells had attached independent of fibronectin precoating. Inclusion of serum in media during the initial period of attachment decreased the number of COS-7 cells attached to SiO(2) surfaces, but no dependence on serum was seen for ITO surfaces. The number of cells attached per unit area varied with the composition of the surface. However, no differences were observed in the percentage of cells transfected with a green fluorescent protein gene, or in the level of reporter gene expression over the population of transfected cells on ITO, SiO(2), Pt, Ag, or Au surfaces. Similar FACS analysis of transfected Hep G2 cells revealed lower levels of both transfection efficiency and levels of GFP fluorescence. Hep G2 cells plated on Ag did not remain attached for analysis, but there were no significant differences between tissue culture plastic and the other biosensor surfaces in the percentage of cells transfected. This suggests that, in general, cells will attach to the various conducting and nonconducting biosensor surfaces studied and will provide comparable data in reporter gene expression assays.
- Runyan, R., Mercado-Pimentel, M. E., & Runyan, R. B. (2007). Multiple transforming growth factor-beta isoforms and receptors function during epithelial-mesenchymal cell transformation in the embryonic heart. Cells, tissues, organs, 185(1-3).More infoEpithelial-mesenchymal cell transformation (EMT) is a critical process during development of the heart valves. Transition of endothelial cells into mesenchymal cells in the atrioventricular (AV) canal and the outflow tract regions of the heart form the cardiac cushions that eventually form the heart valves. Collagen gel invasion assay has aided in the identification of molecules that regulate EMT. Among those, transforming growth factor-beta (TGF-beta) ligands and receptors demonstrate a critical role during EMT. In the chick, TGF-beta ligands and some receptors have specific functions during EMT. TGF-beta2 mediates endothelial cell-cell activation and separation, and TGF-beta3 mediates cell invasion into the extracellular matrix. Receptors involved in the EMT process include TGF-beta receptor type II (TBRII), TBRIII, endoglin and the TBRI receptors, ALK2 and ALK5. In contrast, in the mouse model, TGF-beta2 is the only ligand involved in EMT. The TGF-beta2 null mouse has either increased EMT or a mesenchymal cell proliferation after EMT. However, functional studies of TGF-beta1 in vivo and in vitro showed that TGF-beta1 functions in the EMT of the mouse AV canal. Latent TGF-beta-binding protein (LTBP-1) and endoglin have a role in the EMT process. Therefore, TGF-betas mediate cardiac EMT in both embryonic species. Further studies will reveal the identification of ligand and receptor-specific activities.
- Runyan, R., Mercado-Pimentel, M. E., Hubbard, A. D., & Runyan, R. B. (2007). Endoglin and Alk5 regulate epithelial-mesenchymal transformation during cardiac valve formation. Developmental biology, 304(1).More infoEndoglin is an accessory receptor for TGFbeta and can associate with Alk5 or Alk2. Although prior studies indicated that endoglin and Alk5 were not directly involved in epithelial-mesenchymal transformation (EMT) in the heart, the expression pattern of endoglin prompted a re-examination. We here show that loss of endoglin expression mediated by either antisense DNA or siRNA results in a direct perturbation of EMT and reduced expression of EMT markers including slug, runx2, RhoA, and latrophilin-2. An examination of BrdU incorporation shows that, while endoglin regulates proliferation at an early stage, reduced endothelial cell proliferation does not account for the loss of mesenchyme. As Alk5 interacts with endoglin, we utilized siRNA and a specific inhibitor, HTS466284 (HTS), to perturb this receptor as well. Alk5 inhibition produced similar effects to the inhibition of endoglin. There was a reduction in mesenchymal cell formation and loss of EMT marker expression similar to that seen with endoglin. Alk5 kinase inhibition produced a similar loss of EMT marker expression but showed a contrasting upregulation of the proliferation and remodeling markers, Cyclin B2 and beta-catenin. Alk5 and endoglin both mediate endothelial cell proliferation in younger explants but, by stage 16, loss of endoglin no longer alters proliferation rates. These data show that both Alk5 and endoglin are directly involved in the process of EMT, that they interact with both TGFbeta-regulated activation and invasion pathways and that the roles of these receptors change during cardiac development.
- Rausch, M. P., Hahn, T., Ramanathapuram, L., Bradley-Dunlop, D., Mahadevan, D., Mercado-Pimentel, M. E., Runyan, R. B., Besselsen, D. G., Zhang, X., Cheung, H. -., Lee, W., Ling, L. E., & Akporiaye, E. T. (2006). An Orally Active Small Molecule TGF-beta Receptor I Antagonist Inhibits the Growth of Metastatic Murine Breast Cancer. ANTICANCER RESEARCH, 29(6), 2099-2109.
- Runyan, R., Doyle, S. E., Scholz, M. J., Greer, K. A., Hubbard, A. D., Darnell, D. K., Antin, P. B., Klewer, S. E., & Runyan, R. B. (2006). Latrophilin-2 is a novel component of the epithelial-mesenchymal transition within the atrioventricular canal of the embryonic chicken heart. Developmental dynamics : an official publication of the American Association of Anatomists, 235(12).More infoEndothelial cells in the atrioventricular canal of the heart undergo an epithelial-mesenchymal transition (EMT) to form heart valves. We surveyed an on-line database (http://www.geisha.arizona.edu/) for clones expressed during gastrulation to identify novel EMT components. One gene, latrophilin-2, was identified as expressed in the heart and appeared to be functional in EMT. This molecule was chosen for further examination. In situ localization showed it to be expressed in both the myocardium and endothelium. Several antisense DNA probes and an siRNA for latrophilin-2 produced a loss of EMT in collagen gel cultures. Latrophilin-2 is a putative G-protein-coupled receptor and we previously identified a pertussis toxin-sensitive G-protein signal transduction pathway. Microarray experiments were performed to examine whether these molecules were related. After treatment with antisense DNA against latrophilin-2, expression of 1,385 genes and ESTs was altered. This represented approximately 12.5% of the microarray elements. In contrast, pertussis toxin altered only 103 (0.9%) elements of the array. There appears to be little overlap between the two signal transduction pathways. Latrophilin-2 is thus a novel component of EMT and provides a new avenue for investigation of this cellular process.
- Tavares, A., Mercado-Pimentel, M. E., Runyan, R. B., & Kitten, G. T. (2006). TGF beta-mediated RhoA expression is necessary for epithelial-mesenchymal transition in the embryonic chick heart. DEVELOPMENTAL DYNAMICS, 235(6), 1589-1598.
- Person, A. D., Garriock, R. J., Krieg, P. A., Runyan, R. B., & Klewer, S. E. (2005). Frzb modulates Wnt-9a-mediated beta-catenin signaling during avian atrioventricular cardiac cushion development. DEVELOPMENTAL BIOLOGY, 278(1), 35-48.
- Person, A. D., Klewer, S. E., & Runyan, R. B. (2005). Cell biology of cardiac cushion development. INTERNATIONAL REVIEW OF CYTOLOGY - A SURVEY OF CELL BIOLOGY, VOL 243, 243, 287-+.
- Runyan, R., Person, A. D., Klewer, S. E., & Runyan, R. B. (2005). Cell biology of cardiac cushion development. International review of cytology, 243.More infoThe valves of the heart develop in the embryo from precursor structures called endocardial cushions. After cardiac looping, endocardial cushion swellings form and become populated by valve precursor cells formed by an epithelial-mesenchymal transition (EMT). Endocardial cushions subsequently undergo directed growth and remodeling to form the valvular structures and the membranous septa of the mature heart. The developmental processes that mediate cushion formation include many prototypic cellular actions including adhesion, signaling, migration, secretion, replication, differentiation, and apoptosis. Cushion morphogenesis is unique in that these cellular possesses occur in a functioning organ where the cushions act as valves even while developing into definitive valvular structures. Cardiovascular defects are the most common congenital defects, and one of the most common causes of death during infancy. Thus, there is significant interest in understanding the mechanisms that underlie this complex developmental process. In this regard, substantial progress has been made by incorporating an understanding of cardiac morphology and cell biology with the rapidly expanding repertoire of molecular mechanisms gained through human genetics and research using animal models. This article reviews cardiac morphogenesis as it relates to heart valve formation and highlights selected growth factors, intracellular signaling mediators, and extracellular matrix components involved in the creation and remodeling of endocardial cushions into mature cardiac structures.
- Selmin, O., Thorne, P., Caldwell, P., Johnson, P., & Runyan, R. (2005). Effects of trichloroethylene and its metabolite trichloroacetic acid on the expression of vimentin in the rat H9c2 cell line. CELL BIOLOGY AND TOXICOLOGY, 21(2), 83-95.
- Camenisch, T. D., Molin, D., Person, A., Runyan, R. B., Gittenberger-de, G. A., McDonald, J. A., & Klewer, S. E. (2002). Temporal and distinct TGF beta ligand requirements during mouse and avian endocardial cushion morphogenesis. DEVELOPMENTAL BIOLOGY, 248(1), 170-181.
- Romano, L., & Runyan, R. (2000). Slug is an essential target of TGF beta 2 signaling in the developing chicken heart. DEVELOPMENTAL BIOLOGY, 223(1), 91-102.
- Brown, C., Boyer, A., Runyan, R., & Barnett, J. (1999). Antibodies to the type II TGF beta receptor block cell activation and migration during atrioventricular cushion transformation in the heart. DEVELOPMENTAL BIOLOGY, 174(2), 248-257.
- Romano, L., & Runyan, R. (1999). Slug is a mediator of epithelial-mesenchymal cell transformation in the developing chicken heart. DEVELOPMENTAL BIOLOGY, 212(1), 243-254.
- Runyan, R., Wendler, C., Romano, L., Boyer, A., Dagle, J., & Weeks, D. (1999). Utilization of antisense oligodeoxynucleotides with embryonic tissues in culture. METHODS-A COMPANION TO METHODS IN ENZYMOLOGY, 18(3), 316-324.
- Boyer, A., Ayerinskas, ., Vincent, E., McKinney, L., Weeks, D., & Runyan, R. (1998). TGF beta 2 and TGF beta 3 have separate and sequential activities during epithelial-mesenchymal cell transformation in the embryonic heart. DEVELOPMENTAL BIOLOGY, 208(2), 530-545.
- Boyer, A., Erickson, C., & Runyan, R. (1998). Epithelial-mesenchymal transformation in the embryonic heart is mediated through distinct pertussis toxin-sensitive and TGF beta signal transduction mechanisms. DEVELOPMENTAL DYNAMICS, 214(1), 81-91.
- Collier, J., Selmin, O., Johnson, P., & Runyan, R. (1998). Trichloroethylene effects on gene expression during cardiac development. BIRTH DEFECTS RESEARCH PART A-CLINICAL AND MOLECULAR TERATOLOGY, 67(7), 488-495.
- Pollack, A. L., Runyan, R. B., & Mostov, K. E. (1998). Morphogenetic mechanisms of epithelial tubulogenesis: MDCK cell polarity is transiently rearranged without loss of cell-cell contact during scatter factor hepatocyte growth factor-induced tubulogenesis. DEVELOPMENTAL BIOLOGY, 204(1), 64-79.
- Brown, C., Boyer, A., Runyan, R., & Barnett, J. (1996). Requirement of type III TGF-beta receptor for endocardial cell transformation in the heart. SCIENCE, 283(5410), 2080-2082.
- Boyer, A., & Runyan, R. (1995). TGF beta type III and TGF beta type II receptors have distinct activities during epithelial-mesenchymal cell transformation in the embryonic heart. DEVELOPMENTAL DYNAMICS, 221(4), 454-459.
- Boyer, A., Finch, W., & Runyan, R. (1995). Trichloroethylene inhibits development of embryonic heart valve precursors in vitro. TOXICOLOGICAL SCIENCES, 53(1), 109-117.
- HUANG, J., POTTS, J., VINCENT, E., WEEKS, D., RUNYAN, R., Claycomb, W., & DiNardo, P. (1995). MECHANISMS OF CELL-TRANSFORMATION IN THE EMBRYONIC HEART. CARDIAC GROWTH AND REGENERATION, 752, 317-330.
- CHU, Y. W., RUNYAN, R. B., OSHIMA, R. G., & HENDRIX, M. (1993). EXPRESSION OF COMPLETE KERATIN FILAMENTS IN MOUSE L-CELLS AUGMENTS CELL-MIGRATION AND INVASION. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 90(9), 4261-4265.
- POTTS, J. D., VINCENT, E. B., RUNYAN, R. B., & WEEKS, D. L. (1992). SENSE AND ANTISENSE TGF-BETA-3 MESSENGER-RNA LEVELS CORRELATE WITH CARDIAC-VALVE INDUCTION. DEVELOPMENTAL DYNAMICS, 193(4), 340-345.
- LOEBER, C. P., & RUNYAN, R. B. (1990). A COMPARISON OF FIBRONECTIN, LAMININ, AND GALACTOSYLTRANSFERASE ADHESION MECHANISMS DURING EMBRYONIC CARDIAC MESENCHYMAL CELL-MIGRATION INVITRO. DEVELOPMENTAL BIOLOGY, 140(2), 401-412.
- POTTS, J., & RUNYAN, R. (1990). EPITHELIAL MESENCHYMAL CELL-TRANSFORMATION IN THE EMBRYONIC HEART CAN BE MEDIATED, IN PART, BY TRANSFORMING GROWTH FACTOR-BETA. DEVELOPMENTAL BIOLOGY, 134(2), 392-401.
- POTTS, J., DAGLE, J., WALDER, J., WEEKS, D., & RUNYAN, R. (1990). EPITHELIAL MESENCHYMAL TRANSFORMATION OF EMBRYONIC CARDIAC ENDOTHELIAL-CELLS IS INHIBITED BY A MODIFIED ANTISENSE OLIGODEOXYNUCLEOTIDE TO TRANSFORMING GROWTH-FACTOR BETA-3. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 88(4), 1516-1520.
- RUNYAN, R., POTTS, J., & WEEKS, D. (1990). TGF-BETA-3-MEDIATED TISSUE INTERACTION DURING EMBRYONIC HEART DEVELOPMENT. MOLECULAR REPRODUCTION AND DEVELOPMENT, 32(2), 152-159.
- RUNYAN, R., POTTS, J., SHARMA, R., LOEBER, C., CHIANG, J., & BHALLA, R. (1989). SIGNAL TRANSDUCTION OF A TISSUE INTERACTION DURING EMBRYONIC HEART DEVELOPMENT. CELL REGULATION, 1(3), 301-313.
- BAYNA, E., RUNYAN, R., SCULLY, N., REICHNER, J., LOPEZ, L., & SHUR, B. (1988). CELL-SURFACE GALACTOSYLTRANSFERASE AS A RECOGNITION MOLECULE DURING DEVELOPMENT. MOLECULAR AND CELLULAR BIOCHEMISTRY, 72(1-2), 141-151.
- RUNYAN, R., MAXWELL, G., & SHUR, B. (1986). EVIDENCE FOR A NOVEL ENZYMATIC MECHANISM OF NEURAL CREST CELL-MIGRATION ON EXTRACELLULAR GLYCOCONJUGATE MATRICES. JOURNAL OF CELL BIOLOGY, 102(2), 432-441.
- RUNYAN, R., VERSALOVIC, J., & SHUR, B. (1986). FUNCTIONALLY DISTINCT LAMININ RECEPTORS MEDIATE CELL-ADHESION AND SPREADING - THE REQUIREMENT FOR SURFACE GALACTOSYLTRANSFERASE IN CELL SPREADING. JOURNAL OF CELL BIOLOGY, 107(5), 1863-1871.
- KRUG, E. L., RUNYAN, R. B., & MARKWALD, R. R. (1985). PROTEIN EXTRACTS FROM EARLY EMBRYONIC HEARTS INITIATE CARDIAC ENDOTHELIAL CYTODIFFERENTIATION. DEVELOPMENTAL BIOLOGY, 112(2), 414-426.
- Krug, E. L., Runyan, R. B., & Markwald, R. R. (1985). Protein extracts from early embryonic hearts initiate cardiac endothelial cytodifferentiation. Developmental biology, 112(2), 414-26.More infoPrior to the formation of multiple chambers, the embryonic heart consists of two epithelial tubes, one within the other. As development proceeds, portions of the inner epithelium, i.e., the endothelium, undergo a morphological transformation into a migrating mesenchymal cell population. Our results show that this transformation is affected by proteins secreted by the outer epithelium, i.e., the myocardium, into the extracellular matrix between these two tissues. This conclusion is based on tissue autoradiographic studies of whole embryo cultures with 3H-amino acids. Continuous labeling conditions generated an apparent gradient of proteins extending away from the myocardium and contacting the endothelium just prior to the formation of mesenchyme, i.e., activation of the transformation sequence. Pulse/chase studies confirmed this directional movement of matrix protein. By performing sequential extractions of preactivation staged embryonic hearts with EDTA and testicular hyaluronidase followed by ammonium sulfate precipitation we obtained an enriched preparation of cardiac extracellular matrix. This fraction was capable of eliciting several of the events characteristic of endothelial activation in vitro. These events included: (i) cell-cell separation, (ii) lateral cell mobility, and (iii) hypertrophy and polarization of intracellular PAS staining (Golgi apparati). The biological activity of the extract was sensitive to heat denaturation: a homogenate of the remaining extracted tissue would not substitute for the matrix extract. Morphologically the extracted hearts appeared intact, however, the extracellular matrix space was significantly diminished. No more than 6% of the total lactic dehydrogenase activity, a cytosolic enzyme, was found in the extract. Preliminary electrophoretic characterization of the extract (metabolically labeled with 14C-amino acids) indicated that it may contain as many as 35 proteins or subunits. The relationship of ECM to endothelial differentiation in cardiac morphogenesis is discussed as a model for other developmental systems.
- RUNYAN, R. B., & MARKWALD, R. R. (1983). INVASION OF MESENCHYME INTO 3-DIMENSIONAL COLLAGEN GELS - A REGIONAL AND TEMPORAL ANALYSIS OF INTERACTION IN EMBRYONIC HEART-TISSUE. DEVELOPMENTAL BIOLOGY, 95(1), 108-114.
- Runyan, R. B., & Markwald, R. R. (1983). Invasion of mesenchyme into three-dimensional collagen gels: a regional and temporal analysis of interaction in embryonic heart tissue. Developmental biology, 95(1), 108-14.More infoIn normal heart development the endothelium of the atrioventricular canal, but not the ventricle, produces mesenchymal cells which seed (invade) into the intervening extracellular matrix toward the myocardium at around 64-69 hr of development. We have utilized three-dimensional collagen substrates to examine the initiation of seeding by atrioventricular canal endothelia in vitro and to compare and contrast the responses of the ventricular endothelia. Explants of atrioventricular canals and ventricles from staged embryos were placed on the surfaces of collagen gels prior to the onset of seeding in situ. At varied intervals of incubation, the explant was removed, leaving behind a monolayer on the surface of the gel which consisted of endothelial cells. Subsequently, the endothelial outgrowths were examined for seeded cells. The results confirm the regional endothelial differences seen in vivo. They also show that invasion of the collagen gels is due to an alteration in phenotype mediated by interaction with other components of embryonic heart explant. Lastly, the time course of this tissue interaction in vitro mimics the onset of seeding in vivo.
- Borg, T. K., Runyan, R. B., & Wuthier, R. E. (1981). A freeze‐fracture study of avian epiphyseal cartilage differentiation. Anatomical Record, 199(4), 449-457. doi:10.1002/ar.1091990402
- Markwald, R. R., Krook, J. M., Kitten, G. T., & Runyan, R. B. (1981). Endocardial cushion tissue development: structural analyses on the attachment of extracellular matrix to migrating mesenchymal cell surfaces. Scanning electron microscopy, 261-74.More infoThe progressive growth and eventual fusion of the atrioventricular (AV) endocardial cushions is of critical importance to normal embryonic heart development. Failure to do so would result in septal and AV valvular defects. A central feature in initial cushion growth is the migration of cushion tissue (CT) cells through an heterogeneous extracellular matrix (ECM) which has previously been shown (in particular hyaluronate) to modify migratory behavior. Attention was directed to migrating CT cells to determine if (1) their surfaces physically attach to or bind ECM and (2) are modified to suggest a morphological basis for cell:matrix interaction. The migratory appendages (filopodia) of CT cells maintained in organ culture attached both to collagenous microfibrils coated with polyanionic material and hyaluronate (HA) enriched ECM. The cell:matrix associations were of sufficient strength to restrain the cell from contracting following freezing procedures and were labile to mild trypsin treatment. HA enriched matrix persisted at the cell surface even after treatments which removed most free ECM, but was readily removed by hyaluronidase and trypsin digestion. Freeze fracture analyses revealed 16-18 nm particles elevated above the plane of the filopodial surface which closely interfaced with ECM components. These particles were variably distributed, ranging from almost homogenous dispersion to focalized clusters, but were absent on surrounding non-migratory (myocardial) cells. Results are consistent with a model in which cell attachment to its migratory substratum is mediated by polyanions (probably sulfated glycosaminoglycan and fucosylated glycoprotein) and detachment by hyaluronate.
- Bigger, C. H., & Runyan, R. B. (1979). An in situ demonstration of self-recognition in gorgonians. Developmental & Comparative Immunology, 3, 591-597.
- Seale, T. W., & Runyan, R. B. (1979). Scanning electron microscopy of surface ultrastructure changes during meiosporangium maturation and meiospore liberation in the aquatic fungus Allomycpes arbuscula.. Journal of Bacteriology, 140(1), 276-284.
- Borg, T. K., Runyan, R. B., & Wuthier, R. E. (1978). Correlation of freeze-fracture and scanning electron microscopy of epiphyseal chondrocytes. Calcified Tissue Research, 26(3), 237-241.
- SEALE, T., DELEHANTY, J., & RUNYAN, R. (1974). LIBERATION AND DEVELOPMENT OF ALLOMYCES-ARBUSCULA MITOSPORES VIEWED BY SCANNING ELECTRON-MICROSCOPY. JOURNAL OF BACTERIOLOGY, 120(3), 1417-1426.
- Wang, R., Yun, J. X., Goodwin, R., Markwald, R., Runyan, R. B., Gao, B., Izatt, J., Fujimoto, J., & Tuchin, V. (2013, 2012). Doppler Streak Mode Fourier Domain Optical Coherence Tomography. In OPTICAL COHERENCE TOMOGRAPHY AND COHERENCE DOMAIN OPTICAL METHODS IN BIOMEDICINE XVI, 8213.
- Liu, H., Qin, W., Shao, Y., Wang, Z., Yang, H., Runyan, R. B., Borg, T. K., Gao, B. Z., Farkas, D., Nicolau, D., & Leif, R. (2010, 2013). Disassembly of Myofibrils in Adult Cardiomyocytes during Dedifferentiation. In IMAGING, MANIPULATION, AND ANALYSIS OF BIOMOLECULES, CELLS, AND TISSUES XI, 8587.
- Wang, R., Yun, J. X., Goodwin, R., Markwald, R., Borg, T. K., Runyan, R. B., Gao, B., Kollias, N., Choi, B., Zeng, H., Kang, H., Knudsen, B., Wong, B., Ilgner, J., Izdebski, K., Suter, M., Lam, S., Brenner, M., Gregory, K., , Tearney, G., et al. (2007, 2012). 4D imaging of embryonic chick hearts by streak-mode Fourier domain optical coherence tomography. In PHOTONIC THERAPEUTICS AND DIAGNOSTICS VIII, PTS 1 AND 2, 8207.
- Ma, S., Wang, R., Goodwin, R. L., Markwald, R. R., Borg, T. K., Runyan, R. B., Gao, B. Z., Tuchin, V., Duncan, D., Larin, K., Leahy, M., & Wang, R. (2013, 2013). 4D Display of the Outflow Track of Embryonic-Chick Hearts (HH 14-19) Using a High Speed Streak Mode OCT. In DYNAMICS AND FLUCTUATIONS IN BIOMEDICAL PHOTONICS IX, 8580.
- Runyan, R. B. (2017, Fall 2017). Invited Speaker- "Olfactomedin1 variants differentially suppress or enhance EMT in heart development and metastasis”. TEMTIA & MD Anderson *8th International Conference on EMT. University of Texas MD Anderson Cancer Center, Houston, TX: The Epithelial-Mesenchymal International Association & MD Anderson Cancer Center.
- Runyan, R. B. (2015, February 16). EMT in development and pathology. Cancer Biology Seminar Series. UA Cancer Center: Cancer Biology Program.More infoSeminar presentation in Cancer Center
- Runyan, R. B. (2015, October). Invited Presentation at Meeting Banquet: History and Progress in the Investigation of Epithelial-Mesenchymal Transition. The EMT International Association 7th International Meeting. Melbourne, Australia: The Epithelial Mesenchymal Transition Association.
- Doetschman, T., Barnett, J. V., Runyan, R. B., Camenisch, T. D., Heimark, R. L., Granzier, H. L., Conway, S. J., & Azhar, M. (2013. Transforming growth factor beta signaling in adult cardiovascular diseases and repair(pp 203-223).
- Runyan, R. B., & Savagner, P. (2018, Spring). Editor, Special Issue on EMT in Development, Cancer and Fibrosis. Developmental Dynamics.More infoRecruited authors and solicited both research and review articles for special issue on the Role of EMT in Development, Cancer and Fibrosis. Deadline for issue is March 2017. Papers are all accepted, publication date is March 2018
- Thompson, E. W., Runyan, R., Savagner, P., & Newgreen, D. F. (2013, 2011). Out of the Desert: The 4th TEMTIA Meeting on New Advances in Development, Fibrosis and Cancer Preface. CELLS TISSUES ORGANS.
- Tavares, A. L., & Runyan, R. B. (2012, APR). Temporal and functional study of TGF β regulated genes during chicken AV canal formation. FASEB JOURNAL.
- Runyan, R. B. (2010, APR). Writing for the Reviewers: What are they Looking for?. FASEB JOURNAL.
- Makwana, O., King, N. M., Ahles, L., Granzier, H., Selmin, O., & Runyan, R. B. (2009, MAY). Effects of Trichloroethylene on Heart Development. BIRTH DEFECTS RESEARCH PART A-CLINICAL AND MOLECULAR TERATOLOGY.
- Collier, J., Cover, C., Johnson, P., Selmin, O., & Runyan, R. (2003, MAR 17). Effects of trichloroethylene on gene expression during cardiac development. FASEB JOURNAL.
- HUANG, J., WENDLER, C., & RUNYAN, R. (2001, NOV). CLONING AND EXPRESSION OF MOX1 IN THE CHICKEN-EMBRYO. MOLECULAR BIOLOGY OF THE CELL.
- Runyan, R., Wendler, C., Romano, L., & Boyer, A. (2001, MAR 15). Epithelial-mesenchymal cell transformation of cardiac endothelium is a multistep, transcriptionally regulated, process in the atrioventricular canal. FASEB JOURNAL.
- Boyer, T., Jongewaard, I., Boyer, A., Klewer, S., & Runyan, R. (2000, NOV). Establishment of an in vitro model to examine the effects of gene overexpression in the developing AV canal: Examination of type VI collagen in the developing chick heart. MOLECULAR BIOLOGY OF THE CELL.
- Brown, C., Boyer, A., Runyan, R., & Barnett, J. (2000, OCT 15). Antibodies to the type III transforming growth factor beta receptor block cell activation and migration during atrioventricular cushion transformation in the heart.. CIRCULATION.
- Boyer, A., & Runyan, R. (1999, NOV). Comparative studies of signal transduction in epithelial mesenchymal transformation in the embryonic heart. MOLECULAR BIOLOGY OF THE CELL.
- Boyer, A., Erickson, C., & Runyan, R. (1999, DEC). Further examination of pertussis toxin inhibition of epithelial-mesenchymal transformation in the embryonic chick heart.. MOLECULAR BIOLOGY OF THE CELL.
- BOYER, A., VINCENT, E., WEEKS, D., & RUNYAN, R. (1996, NOV). AN EXAMINATION OF TGF-BETA ISOFORM ACTIVITY DURING EPITHELIAL-MESENCHYMAL CELL-TRANSFORMATION IN THE EMBRYONIC HEART. MOLECULAR BIOLOGY OF THE CELL.
- BROWN, C., BOYER, A., RUNYAN, R., & BARNETT, J. (1996, OCT 15). ANTIBODIES TO THE TYPE-II TRANSFORMING GROWTH-FACTOR-BETA RECEPTOR BLOCK CELL ACTIVATION AND MIGRATION DURING ATRIOVENTRICULAR CUSHION TRANSFORMATION. CIRCULATION.
- HUANG, J., MCKINNEY, L., WENDLER, C., & RUNYAN, R. (1995, SEP). EXAMINATION OF BRACHYURY (T) GENE-EXPRESSION IN THE DEVELOPING AVIAN HEART DURING VALVULAR MESENCHYME FORMATION. MOLECULAR BIOLOGY OF THE CELL.
- RUNYAN, R., POTTS, J., WEEKS, D., SHARMA, R., LOEBER, C., CHIANG, J., BHALLA, R., BOCKMAN, D., & KIRBY, M. (1992, 1990). TISSUE INTERACTION AND SIGNAL TRANSDUCTION IN THE ATRIOVENTRICULAR-CANAL OF THE EMBRYONIC HEART. EMBRYONIC ORIGINS OF DEFECTIVE HEART DEVELOPMENT.
- VINCENT, E., POTTS, J., RUNYAN, R., & WEEKS, D. (1992, SEP). ISOLATION OF AN ENDOGENOUS ANTISENSE MESSENGER-RNA TO TGF-BETA-3 FROM CHICK HEART. MOLECULAR BIOLOGY OF THE CELL.
- WENDLER, C., HUANG, J., MCKINNEY, L., & RUNYAN, R. (1992, SEP). IDENTIFICATION OF BRACHYURY (T) HOMOLOGS IN CHICKEN TISSUES. MOLECULAR BIOLOGY OF THE CELL.
- Wendler, C., & Runyan, R. (1992, MAR 7). The cellular invasion step of epithelial-mesenchymal cell transformation utilizes Mox-1.. FASEB JOURNAL.
- RUNYAN, R., POTTS, J., WEEKS, D., SHARMA, R., LOEBER, C., CHIANG, J., & BHALLA, R. (1991, APR 9). TISSUE INTERACTION AND SIGNAL TRANSDUCTION IN THE ATRIOVENTRICULAR-CANAL OF THE EMBRYONIC HEART. ANNALS OF THE NEW YORK ACADEMY OF SCIENCES.
- RUNYAN, R., MAXWELL, G., & SHUR, B. (1986, 1984). NEURAL CREST MIGRATION ON A COMPLEX EXTRACELLULAR-MATRIX IS INHIBITED BY A MODIFIER OF SURFACE GALACTOSYLTRANSFERASE ACTIVITY. JOURNAL OF CELL BIOLOGY.
- RUNYAN, R., VRSALOVIC, J., & SHUR, B. (1984, NOV). CELL-SURFACE GALACTOSYLTRANSFERASE AS A LAMININ RECEPTOR. JOURNAL OF CELL BIOLOGY.