Spatiotemporal Analysis of CD82-mediated Integrin Adhesion

CD82 介导的整合素粘附的时空分析

• 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
• 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

 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（由申请人提供）：造血干细胞/祖细胞（HSPC）与细胞微环境或“生态位”的相互作用提供基于粘附的信号传导，这对于维持 HSPC 增殖、分化和存活是必要的，尽管使用已取得临床成功。 HSPC 移植用于治疗血癌，调节 HSPC 与其微环境相互作用的机制仍然很大程度上未知，因此，我们确定调节 HSPC 的机制和分子至关重要。该提案的目的是确定 CD82 通过调节整合素聚集和激活来调节整合素介导的 HSPC 粘附的机制。在特定目标 1 中，我将使用蛋白质印迹和流式细胞术分析来量化 β1 激活水平。对配体的响应，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) 来解析总和激活的 β1 整合素的膜蛋白组织 对于特定目标 2，我将使用突变体。 CD82 和原代人 CD34+ 细胞在过继转移实验中评估 CD82 如何调节 HSPC 的归巢和生态位定位 此外，我将使用 SRI 来量化 B1 和 B1 的分子组织。 CD82 在 HSPC 与微环境的接触位点，以确定 CD82 如何调节整合素组织以响应微环境，这一提议具有重要意义，因为我们期望将 CD82 及其调节伙伴确定为增强 HSPC 移植疗法的分子靶点。它将尖端成像技术与体内分析相结合，以提供对四跨膜蛋白如何调节 HSPC 归巢的多尺度理解。此外，我的研究采用了一种独特的方法，针对整合素的上游调节因子， CD82，用于控制 HSPC 归巢 因此，该提案将提供对新型分子调节剂的深入了解，这些调节剂可在移植前增强 HSPC 的粘附特性。