Modulating Cell Phenotype during Tubulogenesis through 3D Micropatterning

通过 3D 微图案调节管发生过程中的细胞表型

基本信息

批准号：
8595863
负责人：
Ryan M Schweller
金额：
$ 4.92万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8595863
关键词：
Active Sites Adhesives Affect Automobile Driving Behavior Biocompatible Materials Biological Blood Vessels Cancerous Cells Characteristics Chemicals Chemistry Complex Cues Custom Development Disease Elements Endothelial Cells Environment Ethylene Glycols Evaluation Event Fibroblast Growth Factor Goals Growth Growth Factor Human Development Hydrogels Individual Lasers Lead Length Ligands Malignant Neoplasms Mechanics Methods Modeling Modification Network-based Nutrient Outcome Oxygen Pattern Pericytes Phenotype Photochemistry Platelet-Derived Growth Factor Play Polyethylene Glycols Procedures Process Property Protocols documentation Relative (related person)Role Scanning Signal Pathway Signal Transduction Signaling Molecule Spatial Distribution Structure Techniques Technology Time Tissues Variant Vascular Endothelial Growth Factors Vascularization Work Wound Healing angiogenesis base cell behavior ethylene glycol extracellular lithography novel therapeutic intervention programs public health relevance response spatiotemporal tumor progression two-photon

项目摘要

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.

描述（由申请人提供）：血管生成是微脉管系统形成中的关键过程，以将营养物和氧气输送到目标细胞和组织。在此过程中，内皮细胞对特定的细胞外信号做出反应，导致它们从现有血管迁移并通过称为肾小管发生的过程形成肾小管。然而，这些信号机制的改变或破坏可能导致不健康血管结构的形成，表明疾病状态（即癌症）。在本提案中，我们的目标是使用微图案生物材料来控制内皮细胞微环境的时空元素，该微环境主要由粘附性、机械性和可扩散/可溶性线索组成。通过观察和表征内皮细胞如何操纵和协调其局部微环境的反应，我们可以根据它们与个体线索的特定相互作用对它们相应的细胞表型和肾小管网络进行分类。为了实现这一目标，我们将首先创建一种基于双光子的图案化策略，能够通过使用正交光化学将多个生物分子并行固定在三维（3D）聚乙二醇（PEG）水凝胶内。此外，我们将整合新的功能，以允许瞬时改变本体水凝胶的特性。利用这项技术，我们将在水凝胶内创建具有大量和局部（图案化）生长因子的粘合剂配体图案。通过首先控制生长因子的空间引入，我们将研究如何在 3D 中控制管发生事件（即分支）的启动。此外，通过使用参与伤口愈合的生长因子：血小板源性生长因子（PDGF）、血管内皮生长因子（VEGF）和成纤维细胞生长因子（FGF），我们将研究每种生长因子遇到的顺序以及个体生长因子的展示（即，大量或局部固定）影响小管网络的关系结构。最后，我们将探讨这些增长的时间引入因子以及它们在肾小管发生过程中的掺入如何改变、破坏或增强内皮细胞反应。从这些研究中，我们预计我们可以控制所形成的小管的分支、伸长和整体结构。为了验证这一点，我们将创建一种综合方法来表征和分类肾小管网络以及内皮细胞本身的表型。最后，空间和时间的变化介绍应该使我们能够解耦这些效应，以创建一个肾小管发生模型，该模型将使我们能够“预编程”3D细胞微环境，以驱动特定的肾小管发生和表型结果。