Spatiotemporal Analysis of CD82-mediated Integrin Adhesion

CD82 介导的整合素粘附的时空分析

基本信息

批准号：
9187854
负责人：
Christina Marie Termini
金额：
$ 1.4万
依托单位：
UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-12-15 至 2017-05-12
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9187854
关键词：
Address Adhesions Adhesives Adoptive Transfer Alpha Cell Animals Biochemical Blood Bone Marrow CD34 gene Cell Adhesion Cell Communication Cell Line Cell Proliferation Cell Transplantation Cell membrane Cells Clinical Data Deposition Extracellular Matrix Family Fibronectins Flow Cytometry Health Heart Hematological Disease Hematopoietic stem cells Homing Human Image Imaging Techniques Immune System Diseases Immune system Integrins KAI1 gene Knowledge Laminin Ligands Liver Maintenance Malignant Neoplasms Mediating Membrane Membrane Proteins Modeling Molecular Molecular Target Mutation Pathologic Patients Physiological Property Proteins Recruitment Activity Regenerative Medicine Regulation Research Resolution Signal Transduction Signaling Molecule Site Spleen Stem cell transplant Stem cells Structure Surface Techniques Testing Transplantation Visual Western Blotting base blood treatment cancer therapy cell motility experimental study improved in vivo innovation insight migration mutant novel protein protein interaction public health relevance receptor response scaffold single molecule spatiotemporal success therapeutic target transplantation medicine

项目摘要

DESCRIPTION (provided by applicant): Hematopoietic stem/progenitor cell (HSPC) interactions with the cellular microenvironment or "niche" provide adhesion based signaling, which is necessary for the maintenance of HSPC proliferation, differentiation, and survival. Despite the clinical successes from using HSPC transplantation for the treatment of blood cancers, the mechanisms that regulate HSPC interactions with their microenvironment remain largely unknown. As such, it is critical that we identify the mechanisms and molecules that regulate HSPC adhesion to the microenvironment to provide clinical targets to enhance HSPC transplantation therapies. The objective of this proposal is to determine the mechanism by which CD82 regulates integrin-mediated HSPC adhesion by modulating integrin clustering and activation. We will test the hypothesis that CD82 regulates HSPC adhesion and homing by modulating integrin clustering and activation. In Specific Aim 1, I will use Western blot and flow cytometry analyses to quantify β1 activation levels in response to ligand to address how CD82 expression and specific CD82 mutations regulate β1 activation. Furthermore, I will use super-resolution imaging (SRI) to resolve the membrane protein organization of total and activated β1 integrins. For Specific Aim 2, I will use mutant CD82 and primary human CD34+ cells in adoptive transfer experiments to assess how CD82 regulates the homing and niche localization of HSPCs. Additionally, I will use SRI to quantify the molecular organization of B1 and CD82 at the HSPC contact site with niche cells to identify how CD82 regulates integrin organization in response to the microenvironment. This proposal is significant because we expect to identify CD82 and its regulatory partners as molecular targets to enhance HSPC transplantation therapies. This proposal is innovative in that it combines cutting-edge imaging techniques with in vivo analyses to provide a multi-scale understanding of how tetraspanins regulate HSPC homing. Moreover, my study takes a unique approach by targeting an upstream regulator of integrins, CD82, for the control HSPC homing. As such, this proposal will provide insight into novel molecular regulators that can be targeted to enhance the adhesive properties of HSPCs prior to transplantation.

描述（由应用提供）：造血干/祖细胞（HSPC）与细胞微环境或“利基”的相互作用提供基于粘合剂的信号，这对于维持HSPC增殖，分化和存活率是必不可少的。尽管使用HSPC移植以治疗血液癌的临床成功，但调节HSPC相互作用与其微环境的机制在很大程度上尚不清楚。因此，至关重要的是，我们必须确定调节对微环境的HSPC粘合剂的机制和分子，以提供临床靶标，以增强HSPC移植疗法。该建议的目的是确定CD82通过调节整合素聚类和激活来调节整合素介导的HSPC粘合剂的机制。我们将通过调节整合素聚类和激活来调节CD82调节HSPC粘合剂和归巢的假设。在特定目标1中，我将使用蛋白质印迹和流式细胞仪分析来量化β1激活水平，以响应配体，以解决CD82表达和特定CD82突变如何调节β1激活。此外，我将使用超分辨率成像（SRI）来解决总和激活的β1整合素的膜蛋白组织。对于特定目标2，我将在自适应转移实验中使用突变体CD82和原代人CD34+细胞来评估CD82如何调节HSPC的归巢和利基定位。此外，我将使用SRI量化HSPC接触位点B1和CD82的分子组织与利基细胞，以识别CD82如何调节整合素组织以响应微环境。该提议很重要，因为我们希望将CD82及其调节伙伴鉴定为增强HSPC移植疗法的分子靶标。该提案具有创新性，因为它结合了尖端成像技术与体内分析，以提供对四叠蛋白如何调节HSPC归位的多尺度理解。此外，我的研究采用了一种独特的方法，可以针对整合素CD82的上游调节剂来控制HSPC归宿。因此，该提案将提供对新型分子调节剂的见解，这些调节剂可以针对移植前增强HSPC的粘合性特性。