VASCULAR BIOLOGY
AT THE UNIVERSITY OF DENVER
Our lab explores how blood vessels function in development and disease.
A single person’s blood vessels will wrap around the earth four times if stretched end-to-end. With this in mind, it is no surprise that blood vessels are absolutely required for survival and are the first organ system to form embryonically. In a process called angiogenesis, endothelial cells, the most basic unit of blood vessels, proliferate, migrate and collectively move to form a complex and expansive vasculature system allowing nutrient penetration to every cell in our bodies. How individual endothelial cells collectively orchestrate such morphogenic feats in a reproducible and stereotyped fashion is unclear. More specifically, how endothelial cells harness intrinsic cytoskeletal programs to move, polarize, communicate with their neighbors and engage their immediate surroundings during angiogenesis is not very well understood, particularly in disease states such as cancer.
To better understand angiogenic processes, our laboratory uses state-of-the-art microscopy techniques and live-imaging, using fluorescently-tagged versions of cytoskeletal regulatory proteins in vitro and in vivo. This allows us to examine cell behavior and cellular processes during dynamic events of morphogenesis, such as lumen formation. In addition to cell culture and organotypic models of angiogenesis, our lab heavily relies on zebrafish as an experimental platform of vascular development. Zebrafish are fantastic model organisms for blood vessel research as they have extra-uterine development and are completely transparent early in life, allowing for imaging of complex morphogenetic events in vivo. Importantly, this model system also allows us to leverage novel CRISPR/Cas9 technologies and other genetic approaches for genomic editing, creation of light-inducible gene expression systems, conditional tissue-specific gene knockouts and other perturbations for dissecting the contribution of various signaling circuits to angiogenesis during development or in disease-related vascular dysfunction.
We are currently exploring how endothelial cells use cell-cell adhesions called tight junctions, to inform rearrangement of the actin and microtubule cytoskeleton for cell polarization during vascular network formation. We are genetically ablating cell contractile machinery components and/or specific Rho GTPase superfamily proteins using CRISPR-based homologous recombination targeting in zebrafish to determine how these program influence junctional integrity and cell polarity. In a separate project, we are investigating how a family of proteins called synaptotagmin-like proteins contribute to lumen formation by helping to specify apical-basal polarity in endothelial cells during lumenigenesis. To do this we are again using genomic targeting techniques such as endogenous protein tagging coupled with advanced light microscopy to best understand the “when” and “where” of synaptotagmin’s involvement in endothelial lumen formation during development. Overall, our research is dedicated to understanding blood vessels form and how these processes go wrong in various disease states for advancement of next-generation therapies.
People
Principal Investigator and lobster enthusiast
Erich Kushner
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2000-04 B.S., Cellular and Molecular Biology - Fort Lewis College - Durango, CO
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2006-10 Ph.D., Integrative Physiology - University of Colorado - Boulder, CO
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2010-16, Postdoctoral Fellow, University of North Carolina - Chapel Hill, NC
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2016-2023, Assistant Professor, University of Denver- Denver, CO
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2023-Present, Associate Professor, University of Denver- Denver, CO
Biological Sciences Ph.D. Candidate
Jasper Farrington
Project 1: ​Characterization of Caveolae
Project 2: EHD2's Role as a Mechanosensor in Endothelial Cells
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Nickname - 'Princess'
Biological Sciences Ph.D. Candidate
Drew Grespin
Project 1: Biological application of micropatterning and image-based computational analysis.
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Project 2: Describing the nuclear role for Slp2.
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Project 3: Long-read isoform sequencing in endothelial cells.
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Nickname - 'Aqua Man'
Biological Sciences Ph.D. Candidate
Joe Capozzi
Project 1: Trafficking mediators of vascular lumen formation
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Project 2: Genetically engineering Danio rerio.
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Nickname- 'The beard'
Biological Sciences Ph.D. Candidate
Maya Kaul
Project: Investigating the role of PDCH1 in Endothelial Cells and Hantavirus invasion.
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Nickname - 'Pun Master'
Biological Sciences Ph.D. Candidate
Talen Niven
Project: ​Influence of Micropatterning on Endothelial Lumen Formation.
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Nickname - 'The Hair'
Undergraduate Researchers
Jordis Bickel - B.S.
Ashley Zavala - B.S.
Carmila Pamintuan - B.S.
Genevieve Springsteen - B.S.
Chris Culkin
Brooke Berenson
Russell Overly
Alumni
Malcolm Kleban PSM - Graduate student
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Aaryn David - Undergraduate researcher
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Patrick Soonthornprapuet - Undergraduate researcher
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Jessica Friedman - Undergraduate researcher
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Jack Kurlinski - Undergraudate researcher
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Caitlin Francis Ph.D. - Graduate studnet
Current: Postdoctoral Fellow, Yale
Makenzie Bell- Undergraduate researcher
Marina Skripnichuck- Undergraduate researcher
Rachael Judson- Masters student
Stephen Gross Ph.D.- Graduate student
Amelia Webb- Masters student
Current- Idaho College of Osteopathic Medicine
Halye Kincross- Former undergraduate researcher
Current: BC Cancer Research Institute, graduate student
Alek Peterlin- Former undergraduate researcher
Current: University of Utah, Ph.D. candidate
Shae Claflin- Former undergraduate researcherCurrent: University of Colorado, Medical student
PUBLICATIONS
Grespin DB, Niven TG, Babson RO, Kushner EJ. Lipidure-based micropattern fabrication for stereotyping cell geometry. Sci Rep. 2023 Nov 22;13(1):20451. doi: 10.1038/s41598-023-47516-8. PMID: 37993505; PMCID: PMC10665372.
Francis CR, Bell ML, Skripnichuk MM, Kushner EJ. Arf6 is required for endocytosis and filamentous actin assembly during angiogenesis in vitro. Microcirculation. 2023 Nov;30(8):e12831. doi: 10.1111/micc.12831. Epub 2023 Sep 26. PMID: 37750425; PMCID: PMC10688150.
Francis CR, Bell ML, Skripnichuk MM, Kushner EJ. Arf6 Regulates Endocytosis and Angiogenesis by Promoting Filamentous Actin Assembly. bioRxiv [Preprint]. 2023 Feb 22:2023.02.22.529543. doi: 10.1101/2023.02.22.529543. Update in: Microcirculation. 2023 Nov;30(8):e12831. doi: 10.1111/micc.12831. PMID: 36865161; PMCID: PMC9980066.
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Francis CR, *Kincross H, Kushner EJ. Rab35 governs apicobasal polarity through regulation of actin dynamics during sprouting angiogenesis. Nat Commun. 2022 Sep 8;13(1):5276. doi: 10.1038/s41467-022-32853-5. PMID: 36075898.
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Francis CR, Kushner EJ. Trafficking in blood vessel development. Angiogenesis. 2022 Aug;25(3):291-305. doi: 10.1007/s10456-022-09838-5. Epub 2022 Apr 21. Review. PubMed PMID: 35449244
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Webb AM, Francis CR, Judson RJ, Kincross H, Lundy KM, Westhoff DE, Meadows SM, Kushner EJ. EHD2 modulates Dll4 endocytosis during blood vessel development. Microcirculation. 2021. PMID: 34820962
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Francis, C.R. and E.J. Kushner, Capturing membrane trafficking events during 3D angiogenic development in vitro. Microcirculation, 202. PMID: 34415654
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Gross SJ, Webb AM, Peterlin AD, Durrant JR, Judson RJ, Raza Q, Kitajewski JK, Kushner EJ. Notch Regulates Vascular Collagen IV Basement Membrane Through Modulation of Lysyl Hydroxylase 3 Trafficking. Angiogenesis. 2021. PMID: 33956260
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Francis CR, Claflin S, Kushner EJ. Synaptotagmin-Like Protein 2a Regulates Angiogenic. Lumen Formation via Weibel-Palade Body Apical Secretion of Angiopoietin-2. Arterioscler Thromb Vasc Biol. 2021. PMID: 33853352.
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Payne LB, Darden J, Suarez-Martinez AD, Zhao H, Hendricks A, Hartland C, Chong D, Kushner EJ, Murfee WL, Chappell JC. Pericyte migration and proliferation are tightly synchronized to endothelial cell sprouting dynamics.
Integr Biol (Camb). PMID: 33515222
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Buglak DB, Kushner EJ, Marvin AP, Davis KL, Bautch VL. Excess centrosomes disrupt vascular lumenization and endothelial cell adherens junctions. Angiogenesis. 2020. PMID: 32699963
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Yu Z, Mouillesseaux KP, Kushner EJ, Bautch VL. Tumor-Derived Factors and Reduced p53 Promote Endothelial Cell Centrosome Over-Duplication. PLoS One. 2016. PMID: 2797777
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Mouillesseaux K., Wiley D., Saunders L., Wylie L., Kushner E., Chong D., Citrin K., Barber A., Park Y., Kim J., Samsa L.A., Kim J., Liu J., Jin W., Bautch V. Notch Regulates BMP Responsiveness and Lateral Branching in Vessel Networks via SMAD6. Nature Communications. 2016. PMID: 27834400
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Kushner EJ, LS Ferro, Z Yu and VL. Bautch. Excess Centrosomes Perturb Dynamic Endothelial Cell Repolarization During Blood Vessel Formation. Mol Biol Cell. 2016. PMID:27099371
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Wright CE, EJ Kushner, Q Du, and VL Bautch. LGN Directs Interphase Endothelial Cell Behavior via the Microtubule Network. PLoS One. 2015. PMID:26398908
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KR. Klein, NO Karpinich, ST Espenshied, HH Willcockson, WP Dunworth, SL. Hoopes, EJ Kushner, VL Bautch, and KM Caron. Decoy Receptor CXCR7 Modulates Adrenomedullin-Mediated Cardiac and Lymphatic Vascular Development. Dev Cell. 2014. PMID:25203207
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Kushner EJ, LS Ferro, JY Liu, JR Durrant, SL Rogers, AC Dudley, VL Bautch. Excess centrosomes disrupt endothelial cell migration via centrosome scattering. J Cell Biol. 2014. PMID:25049273
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Charpentier MS, Christine KS, Amin NM, Dorr KM, Kushner EJ, Bautch VL, Taylor JM, Conlon FL. CASZ1 promotes vascular assembly and morphogenesis through the direct regulation of an EGFL7/RhoA-mediated pathway. Dev Cell. 2013. PMID:23639441
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Kushner EJ, Bautch VL. Building blood vessels in development and disease. Curr Opin Hematol. 2013. PMID:23567339
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Diehl KJ, Weil BR, Westby CM, MacEneaney OJ, Kushner EJ, Greiner JJ, DeSouza CA. Effects of Endothelin-1 on Endothelial Progenitor Cell Function. Clin Chem Lab Med. 2012. PMID:22706256
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Weil BR, Kushner EJ, Diehl KJ, Greiner JJ, Stauffer BL, DeSouza CA. CD31+ T cells, Endothelial Function and Cardiovascular Risk. Heart Lung Circ. 201.
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Kushner EJ, Weil BR, MacEneaney OJ, Morgan RG, Mestek ML, Van Guilder GP, Diehl KJ, Stauffer BL, DeSouza CA.Human Aging and CD31+ T-cell Number, Migration, Apoptotic Susceptibility, and Telomere Length. J Appl Physiol. 2010. PMCID: PMC3006402
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MacEneaney OJ, DeSouza CA, Weil BR, Kushner EJ, Van Guilder GP, Mestek ML, Greiner JJ, Stauffer BL. Prehypertension and Endothelial Progenitor Cell Function. J Hum Hypertens. 2011. PMCID: PMC:2895004
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MacEneaney OJ, Kushner EJ, Westby CM, Cech JN, Greiner JJ, Stauffer BL, DeSouza CA. Endothelial Progenitor Cell Function, Apoptosis, and Telomere Length in Overweight/Obese Humans. Obesity (Silver Spring). 2010. PMCID: PMC2643316
Kushner E, GL Hoetzer, OJ MacEneaney, JJ Greiner, JN Cech, BL Stauffer, CA DeSouza. Aging and Endothelial Progenitor Cell Release of Proangiogenic Cytokines. Age Ageing. 2010. PMCID: PMC2842112
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Klawitter J, J Klawitter, E Kushner, KR Jonscher, J Bendrick-Peart, U Christians, V Schmitz, Association of Immunosuppressant-Induced Protein Changes in the Rat Kidney with Changes in Urine Metabolite Patterns: A Proteo-Metobonomic Study. J Proteome Res. 2010.
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Kushner EJ, RG Morgan, AM Van Engelenburg, OJ MacEneaney, GP Van Guilder, CA DeSouza. CD31+ T Cells Represent a Functionally Distinct Vascular T Cell Phenotype. Blood Cells, Molecules and Diseases. 2010.
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MacEneaney OJ, EJ Kushner, GP Van Guilder, JJ Greiner, BL Stauffer, CA DeSouza. Endothelial Progenitor Cell Number and Colony-Forming Capacity in Overweight and Obese Adults. International Journal of Obesity. 2009. PMC2643316
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Kushner EJ, GP Van Guilder, OJ MacEneaney, JN Cech, AJ Zaug, BL Stauffer, CA DeSouza. Aging and Endothelial Progenitor Cell Telomere Length in Healthy Men. Clinical Chemistry and Laboratory Medicine. 2009. PMC2646422
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Stauffer BL, OJ MacEneaney, EJ Kushner, JN Cech, JJ Greiner, CM Westby, CA DeSouza. Gender and Endothelial Progenitor Cell Number in Middle-Aged Adults. Artery Research. 2008. PMCID: PMC2636711
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Stauffer BL, Kushner EJ, Wulfman T, Zeller T, Sobus R, Westby CM. Transcriptional Regulation of Beta2-Microglobulin Demonstrated via a Novel Genomic and Proteomic Analysis of Percutaneously Collected Peripheral Atheroma. Clin Transl Sci. 2008. PMCID: PMC20443854
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Stauffer BL, R Sobus, EJ Kushner, CM Westby T Wulfman, T Zeller. Feasibility of Molecular Analysis of Percutaneously Collected Peripheral Atheroma. Excerpta Medica Inc-Elsevier Science Inc, 2007.
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GALLERY
POSITIONS
PROSPECTIVE GRADUATE
STUDENT SCIENTISTS
Inquisitive minds who are interested in conducting research as a career look in our masters and PhD programs in biology.
CELLULAR
BIOPHYSICS PROGRAM
I highly recommend this program as it grants students access to multiple departments (Chemistry, Biochemistry, Biology, Physics) for first year rotations and its curriculum is specifically geared towards interdisciplinary training.
PROSPECTIVE POSTDOCTORAL SCHOLARS
Highly motivated individuals are encouraged to contact Dr. Kushner directly.
CONTACT
Erich Kushner, Ph.D.
Associate Professor
Molecular and Cellular Biophysics Program
Dept. of Biological Sciences
University of Denver