Analysis of mDia formins in hematopoietic stem cell engraftment and migration

mDia 福明在造血干细胞植入和迁移中的分析

• 批准号: 7872318
• 负责人: Peng Ji
• 金额: $ 9万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7872318
• 关键词: Acetylation; Actins; Affect; Aplastic Anemia; Area; Arts; Backcrossings; Binding Proteins; Biochemical; Biological Assay; Biological Sciences; Blood; Blood Cells; Bone Marrow; Bone Marrow Transplantation; Burn injury; C57BL/6 Mouse; Cell Adhesion; Cell Lineage; Cells; Cellular biology; Clinical; Co-Immunoprecipitations; Collaborations; Complex; Cooley' s anemia; Cytoskeletal Proteins; Cytoskeleton; Data; Deacetylase; Deacetylation; Disease; Doctor of Philosophy; Engraftment; Environment; Erythroblasts; F-Actin; Family; Fanconi' s Anemia; Fetal Liver; Fibronectins; Generations; Goals; Guanosine Triphosphate Phosphohydrolases; HDAC6 gene; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Histones; Homing; Immunologic Deficiency Syndromes; In Vitro; Institutes; Kinetics; Knockout Mice; Laboratories; Light; Mammals; Mass Spectrum Analysis; Mediating; Medicine; Mentors; Mentorship; Microfilaments; Microscopy; Migration Assay; Modification; Molecular; Molecular Biology; Multiple Myeloma; Mus; Myosin Type II; Patients; Phosphorylation; Play; Post-Translational Protein Processing; Process; Protein Family; Proteins; Publishing; Regulation; Research; Research Personnel; Role; Scientist; Sickle Cell Anemia; Signal Pathway; Singapore; Stem Cell Research; Stem cells; T-Lymphocyte; Techniques; Time; TimeLine; Training; Transplantation; Umbilical Cord Blood; Wasps; Work; beta Thalassemia; career; cell motility; cell type; college; embryonic stem cell; experience; hematopoietic tissue; in vivo; interest; leukemia/lymphoma; loss of function; macrophage; migration; mouse model; novel; polymerization; public health relevance; research study; skills; stem cell population; vector

DESCRIPTION (provided by applicant): I am a MD/PhD postdoctoral scientist with extensive training in molecular and cell biology from Albert Einstein College of Medicine and Whitehead Institute. My research focuses on the roles of the mDia formin proteins on hematopoietic cells. The mDia formin proteins have well-established roles on the migration of many different cell types. However, their effects on the migration of hematopoietic cells, especially hematopoietic stem cells (HSCs), are poorly understood. With the ideal research environment of Whitehead Institute and excellent mentorship of Dr. Harvey Lodish, I plan to expand my existing scientific and intellectual skills by developing expertise in areas of hematopoeitic stem cells and cell migration, which is vital to my long term career goal of becoming a successful, independent investigator. The goal of my proposed study is to characterize the functional roles of mDia formins, specifically mDia1 and mDia2, on hematopoietic stem cell (HSC) engraftment and migration. HSC engraftment and migration are critical for successful bone marrow transplantation, which has become a routine clinical strategy for treating patients with many blood related diseases such as aplastic anemia, fanconi anemia, sickle cell anemia, beta thalassemia major, leukemia, lymphomas, multiple myeloma, and many immune deficiency disorders. The intracellular signaling pathways regulating HSC engraftment and migration often directly or indirectly target on the actin cytoskeleton network through proteins that regulate polymerization of the actin proteins. The mDia formin proteins are a major actin-nucleating family and their functions in HSCs are poorly understood. To elucidate the roles of mDia formins, specifically mDia1 and mDia2, on HSC migration, genetically modified mouse models and molecular cell biology techniques will be used. Specifically, mouse models deficient of mDia1 or mDia2 will be generated. With these mice, bone marrow and fetal liver Lin-Sca1+Kit+ (LSK) cells, an enriched HSC population, will be purified to perform in vitro migration assays, short-term homing, and long- term engraftment analysis. In collaboration with Drs. Tzutzuy Ramirez Hernandez and Maki Murata-Hori of the Temasek Life Sciences Laboratory in Singapore, the kinetic interactions of mDia1 and mDia2 with actin cytoskeletal proteins and how these interactions regulate hematopoietic stem cell migration will be investigated using confocal fluorescent microscopy and mass spectrometry analysis. In addition, post-translational modifications of mDia1 and mDia2, especially acetylation and phosphorylation, and how these modifications affect their functions in HSC migration will also be characterized. PUBLIC HEALTH RELEVANCE: Current clinical and experimental HSC transplantation is still facing two obstacles: the limited number of transplantable HSCs in a bone marrow or cord blood unit; and a low efficiency of engraftment of transplanted HSCs into recipient bone marrow niches. The project proposed here is directly relevant to these issues. Successful accomplishment of this proposed study will help elucidate how mDia formins mediated regulation of the actin cytoskeleton network affects functions of hematopoietic stem cells. Understanding these processes is highly relevant for bone-marrow transplantation therapies to treat blood-related diseases.

描述（由申请人提供）：我是一名MD/PhD博士后科学家，在阿尔伯特爱因斯坦医学院和怀特黑德研究所接受了广泛的分子和细胞生物学培训。我的研究主要集中在mDia蛋白对造血细胞的作用。mDia β蛋白在许多不同细胞类型的迁移中具有公认的作用。然而，它们对造血细胞，特别是造血干细胞（HSC）迁移的影响知之甚少。在怀特黑德研究所的理想研究环境和哈维·洛迪什博士的出色指导下，我计划通过发展造血干细胞和细胞迁移领域的专业知识来扩展我现有的科学和智力技能，这对我成为一名成功的独立研究者的长期职业目标至关重要。 我提出的研究的目标是表征mDia formins，特别是mDia 1和mDia 2，对造血干细胞（HSC）植入和迁移的功能作用。造血干细胞的植入和迁移是骨髓移植成功的关键，骨髓移植已成为治疗许多血液相关疾病患者的常规临床策略，如再生障碍性贫血、范可尼贫血、镰状细胞贫血、重型β地中海贫血、白血病、淋巴瘤、多发性骨髓瘤和许多免疫缺陷疾病。调节HSC植入和迁移的细胞内信号传导途径通常通过调节肌动蛋白聚合的蛋白质直接或间接靶向肌动蛋白细胞骨架网络。mDia β蛋白是一个主要的肌动蛋白成核家族，其在HSC中的功能知之甚少。为了阐明mDia formins，特别是mDia 1和mDia 2对HSC迁移的作用，将使用遗传修饰的小鼠模型和分子细胞生物学技术。具体而言，将生成mDia 1或mDia 2缺陷的小鼠模型。对于这些小鼠，将纯化骨髓和胎肝Lin-Sca 1 +Kit+（LSK）细胞（富集的HSC群），以进行体外迁移试验、短期归巢和长期植入分析。与新加坡淡马锡生命科学实验室的Tzutzuy Ramirez埃尔南德斯和Maki Murata-Hori博士合作，将使用共聚焦荧光显微镜和质谱分析研究mDia 1和mDia 2与肌动蛋白细胞骨架蛋白的动力学相互作用以及这些相互作用如何调节造血干细胞迁移。此外，还将表征mDia 1和mDia 2的翻译后修饰，特别是乙酰化和磷酸化，以及这些修饰如何影响其在HSC迁移中的功能。 公共卫生关系：目前临床和实验HSC移植仍然面临两个障碍：骨髓或脐带血单位中可移植的HSC数量有限;移植的HSC植入受体骨髓龛的效率低。这里提出的项目与这些问题直接相关。这项研究的成功完成将有助于阐明mDia formins介导的肌动蛋白细胞骨架网络的调节如何影响造血干细胞的功能。了解这些过程与骨髓移植治疗血液相关疾病高度相关。