Functional Role of Tetraspanin CD82 in Hematopoietic Stem Cell Interaction

四跨膜蛋白 CD82 在造血干细胞相互作用中的功能作用

• 批准号: 10001447
• 负责人: Jennifer Gillette
• 金额: $ 37.88万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10001447
• 关键词: Adhesions; Adhesives; Adoptive Transfer; Behavior; Biochemical; Biological; Blocking Antibodies; Blood; Bone Marrow; Cell Adhesion; Cell Communication; Cell Separation; Cell Transplantation; Cell physiology; Cells; Clinic; Clinical; Cues; Data; Defect; Engraftment; Equilibrium; Evaluation; Goals; Health; Hematopoietic Neoplasms; Hematopoietic Stem Cell Mobilization; Hematopoietic System; Hematopoietic stem cells; High Dose Chemotherapy; Home environment; Homing; Human; Image; Imaging technology; Immunologic Deficiency Syndromes; Instruction; Integrins; KAI1 gene; Knockout Mice; Knowledge; Measurement; Measures; Mediating; Membrane; Molecular; Morphology; Mus; Mutation Analysis; Pancytopenia; Patients; Post-Translational Protein Processing; Property; Proteins; Regenerative Medicine; Regulation; Research; Resolution; Role; Sampling; Scaffolding Protein; Signal Transduction; Site; Stem cell transplant; Testing; Transplantation; Work; base; blood treatment; combinatorial; density; expectation; experience; experimental study; extracellular; imaging modality; improved; in vivo; innovation; migration; molecular scale; mouse model; nanoscale; new therapeutic target; novel; public health relevance; self-renewal; single molecule; stem cells; success; therapeutic target; trafficking; transplantation medicine

 DESCRIPTION (provided by applicant): Hematopoietic stem/progenitor cell (HSPC) communication with the cellular microenvironment is critical for the regulation of stem cell functions. Despite their essential role in the clinical setting, our current understanding of the molecular cues that regulate the trafficking of HSPCs and their niche interactions remain rudimentary. Therefore, to improve HSPC transplantation as a treatment option for patients, it is critical that we identify the molecules and mechanisms that regulate HSPC adhesion, trafficking and repopulation. The objective of this proposal is to determine how the molecular scaffold protein, CD82, regulates HSPC localization and adhesive interactions with the bone marrow, which directly influences HSPC function. We will test the hypothesis that CD82 modulates HSPC homing, mobilization, and long-term repopulation capacity through the regulation of integrin clustering and niche adhesion. In Specific Aim 1, we will use our experience with super-resolution imaging methods and primary human cells to quantify the molecular distribution of CD82 membrane organization and determine the effects on integrin clustering, cell adhesion and homing. For Specific Aim 2, we will determine how changes in CD82 expression alter bone marrow retention, homing, and long-term repopulation of HSPCs using the CD82KO mice. This contribution is significant because we expect to identify CD82 as a novel therapeutic target to improve HSPC isolations and transplant efficacy in the clinic. Furthermore, the combination of quantitative single molecule and in vivo information that we propose to obtain has not been measured previously and will bring new perspectives to the function of CD82 in HSPC adhesion/signaling. As such, this proposal is innovative because it will apply a combinatorial, experimental approach to the problem of HSPC adhesion, bone marrow trafficking and repopulation. By utilizing mouse models, mutational analysis, super-resolution imaging, and primary patient samples, we will integrate molecular, biochemical and morphological information to obtain a multi-scale understanding of the role of CD82 in regulating HSPC/niche interactions and the a4ß1 integrin.

 描述(申请人提供)：造血干/祖细胞(HSPC)与细胞微环境的通讯对干细胞功能的调节至关重要。尽管它们在临床环境中扮演着重要的角色，但我们目前对调控HSPC运输的分子线索及其利基相互作用的了解仍然处于初级水平。因此，为了改善HSPC移植作为患者的治疗选择，我们必须确定调控HSPC黏附、运输和再繁殖的分子和机制。这个建议的目的是确定分子支架蛋白CD82如何调节HSPC的定位和与骨髓的黏附相互作用，这直接影响HSPC的功能。我们将验证CD82通过调节整合素聚集和生态位黏附来调节HSPC归巢、动员和长期再繁殖能力的假设。在具体目标1中，我们将利用我们在超分辨率成像方法和原代人类细胞方面的经验来量化CD82膜组织的分子分布，并确定其对整合素聚集、细胞黏附和归巢的影响。对于特定的目标2，我们将使用CD82KO小鼠来确定CD82表达的变化如何改变HSPC的骨髓保留、归巢和长期再繁殖。这一贡献意义重大，因为我们希望将CD82确定为一种新的治疗靶点，以改善HSPC的分离和临床移植疗效。此外，我们建议获得的定量单分子和体内信息的组合以前没有被测量过，这将为CD82在HSPC黏附/信号转导中的功能带来新的视角。因此，这项建议是创新的，因为它将应用一种组合的实验方法来解决HSPC黏附、骨髓贩运和再繁殖的问题。通过利用小鼠模型、突变分析、超分辨率成像和原始患者样本，我们将整合分子、生化和形态信息，以获得对CD82在调节HSPC/NICE相互作用和A4？1整合素中的作用的多尺度理解。