Modulating Cell Phenotype during Tubulogenesis through 3D Micropatterning


Principal Investigator: Ryan M Schweller
Abstract: DESCRIPTION (provided by applicant): Angiogenesis is a critical process in the formation of microvasculature to deliver nutrients and oxygen to target cells and tissues. During this process, endothelial cells respond to specific extracellular signals that cause them to migrate from existing vessels and form tubules through a process called tubulogenesis. Alterations or disruptions in these signaling mechanisms, though, can lead to the formation of unhealthy vessel structures, indicative of disease states (i.e. cancers). In this proposal, we aim to use micropatterned biomaterials to control the spatiotemporal elements of endothelial cell microenvironments, composed primarily of adhesive, mechanical, and diffusible/soluble cues. By observing and characterizing how endothelial cells manipulate and coordinate responses from their local microenvironment, we can classify their corresponding cellular phenotypes and tubule networks based upon their specific interactions with individual cues. To accomplish this, we will first create a two-photon-based patterning strategy capable of immobilizing multiple biomolecules in parallel within three dimensional (3D) poly(ethylene glycol) (PEG) hydrogels through the use of orthogonal photochemistries. In addition, we will incorporate new functionalities to allow the bulk hydrogel properties to be transiently modified. Using this technology we will create patterns of adhesive ligands within the hydrogel with bulk and localized (patterned) growth factors. By first controlling the spatial introduction of growth factos, we will investigate how we can control the initiation of tubulogenic events (i.e. branching) in 3D. Furthermore, by employing the growth factors involved in wound healing: platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), and fibroblast growth factor (FGF), we will investigate how the order in which each growth factor is encountered as well as the display (i.e., bulk or locally immobilized) of the individual growth factors effects the relatie structure of the tubule network. Finally, we will explore the temporal introduction of these growth factors and how their incorporation during tubulogenesis can alter, disrupt, or reinforce endothelial cell responses. From these studies, we anticipate that we can control the branching, elongation, and overall structure of the tubules that are formed. To verify this we will create a comprehensive method to characterize and classify tubule networks as well as the phenotype of the endothelial cells, themselves. Finally, the variations in the spatial and temporal introduction should enable us to decouple these effects to create a tubulogenic model which will allow us to "pre-program" 3D cellular microenvironments to drive specific tubulogenic and phenotypic outcomes.
Funding Period: 2013-09-01 - 2016-08-31
more information: NIH RePORT

Detail Information

Research Grants30

  1. Four-Dimensional Heterogeneity of Fluid Phase Biopsies in Cancer (4DB-Center)
    Peter Kuhn; Fiscal Year: 2013
    ..The 4DB Center will also serve to disseminate information, education, and training to a new generation of cancer physicists;a generation that will implement the power of physics to conquer the problems of cancer. ..
  2. Hyaluronan Matrices in Vascular Pathologies
    Vincent C Hascall; Fiscal Year: 2013
    ..abstract_text> ..
    Kenneth H Cowan; Fiscal Year: 2013
  4. Targeting epithelial cells to treat pulmonary fibrosis
    Dean Sheppard; Fiscal Year: 2013
    ..abstract_text> ..
  5. Fabrication, perfusion and imaging of pre-vascularized tissue scaffolds
    Jennifer L West; Fiscal Year: 2013
    ..Research goals include the optimization of hydrogel scaffold design to deliver growth factors and cells and the characterization of the host response to guide new scaffold designs. ..
  6. Chemotactic Signal Transduction
    Tobias Meyer; Fiscal Year: 2013
    ..Our overall goal in this grant is to understand the key molecular and cellular mechanisms of how single cells and groups of human cells migrate. ..
  7. Center for Excellence in Diabetes and Obesity Research
    ..Continued support to the Center will strengthen the infrastructure of biomedical research at the University of Louisville and will positively impact the field of diabete and obesity research worldwide. ..
  8. Using micropost arrays to measure traction forces during dendritic cell motility
    Daniel A Hammer; Fiscal Year: 2013
    ..abstract_text> ..
  9. Mechanism of Neuropilin Dependent Angiogenesis
    CRAIG VANDER KOOI; Fiscal Year: 2013
    ..Taken together, these studies will provide a molecular understanding of the physical mechanisms underlying the critical first step connecting VEGF ligand binding to receptor activation. ..
  10. The Biology of Prostate Cancer Skeletal Metastases
    EVAN TODD KELLER; Fiscal Year: 2013
    ..This combination of investigators, projects and cores result in a highly synergistic Program that will continue to provide cutting-edge research on PCa bone metastases. ..
  11. Regulation and Function of the Matricellular Protein CCN1 in Ischemic Retinopathy
    Brahim Chaqour; Fiscal Year: 2013
    ..This proposal will provide new insights into the molecular mechanisms of CCN1 activities in vivo and may foster future safer, less destructive, and more effective therapies to harness ischemia-induced neovascularization in ROP. ..
  12. Myeloid Vascular Endothelial Growth Factor Expression &its Role in Tumorigenesis
    RANDALL SCOTT JOHNSON; Fiscal Year: 2013
    ..PUBLIC HEALTH RELEVANCE: These findings speak directly to breast cancer therapies currently used in the clinic, and will explore ways in which these therapies can be improved through modulation of inflammatory response. ..
  13. The Biology of Perlecan in Cancer and Angiogenesis
    Renato V Iozzo; Fiscal Year: 2013
  14. Local regulation of angiogenesis by microenvironment
    Christopher S Chen; Fiscal Year: 2013
  15. Probing biochemical/biophysical influences on endothelial-mesenchymal transition
    MICHAEL PAUL SCHWARTZ; Fiscal Year: 2013
    ..Further, understanding how synthetic biomaterials influence EndMT could ultimately have relevance to medical implant design, particularly in scenarios in which it is important to limit fibrosis or heterotopic ossification. ..
    Harvey R Herschman; Fiscal Year: 2013
  17. New Approaches To Cardiothoracic Tolerance Induction
    Joren C Madsen; Fiscal Year: 2013
    ..We anticipate ongoing progress will continue to contribute to a reduction in the morbidity and mortality associated with solid organ transplantation. ..
  18. Cell Instructive Materials For Engineering Vascular Grafts
    Robert E Akins; Fiscal Year: 2013
  19. Structure and Function of DNA Repair Enzymes
    Susan S Wallace; Fiscal Year: 2013
    ..In addition to bioinformatics for all projects, Core A will also perform kinetics analysis for Projects 2-4. Core C will provide the administrative underpinnings for the project. ..
  20. Generating Vascular Graft Luminal and Medial Layers Based on Multipotent Stem Cel
    Mariah S Hahn; Fiscal Year: 2013
    ..AIM 2: Identify growth factor-laden, PEG "cement" formulations that promote ASC differentiation into EC-like phenotypes. ..
  21. Regulation of CNS viral persistence
    Cornelia Bergmann; Fiscal Year: 2013
    ..Importantly, it will provide valuable information on the interactions of specific CNS cells involved in viral persistence and demyelination and the cellular and soluble mediators of the host immune response...