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: 8532962
• 负责人: Peng Ji
• 金额: $ 23.15万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 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 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8532962
• 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; 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; 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; research study; skills; stem cell population; vector

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.

我是一名医学博士/博士博士后科学家，在阿尔伯特大学接受了广泛的分子和细胞生物学培训。 爱因斯坦医学院和怀特黑德研究所。我的研究集中在mDia的作用， 造血细胞上的蛋白质。mDia β蛋白在许多细胞的迁移中具有公认的作用， 不同的细胞类型然而，它们对造血细胞迁移，特别是造血干细胞迁移的影响， 干细胞（HSC）的研究很少。怀特海研究所拥有理想的研究环境， 在哈维·洛迪什博士的出色指导下，我计划通过以下方式扩展我现有的科学和智力技能： 发展造血干细胞和细胞迁移领域的专业知识，这对我的长期工作至关重要。 成为一名成功的独立调查员的职业目标。 我提出的研究的目标是表征mDia formins的功能作用，特别是mDia 1 和mDia 2对造血干细胞（HSC）植入和迁移的影响。HSC植入和迁移是 成功的骨髓移植是至关重要的，这已经成为治疗的常规临床策略。 患有许多血液相关疾病的患者，如再生障碍性贫血、范可尼贫血、镰状细胞性贫血、β 重型地中海贫血、白血病、淋巴瘤、多发性骨髓瘤和许多免疫缺陷疾病。的 调控HSC植入和迁移的细胞内信号传导途径通常直接或间接靶向 通过调节肌动蛋白聚合的蛋白质形成肌动蛋白细胞骨架网络。mDia α-肌动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。到 阐明mDia formins，特别是mDia 1和mDia 2，对HSC迁移，遗传修饰， 将使用小鼠模型和分子细胞生物学技术。具体地说，缺乏 将生成mDia 1或mDia 2。用这些小鼠，骨髓和胎肝Lin-Sca 1 +Kit+（LSK）细胞， 富集的HSC群体，将被纯化以进行体外迁移测定、短期归巢和长期归巢。 术语植入分析。与联合国儿童基金会的Tzutzuy Ramirez埃尔南德斯和Maki Murata-Hori博士合作， 新加坡淡马锡生命科学实验室，mDia 1和mDia 2与肌动蛋白的动力学相互作用 细胞骨架蛋白以及这些相互作用如何调节造血干细胞迁移将被研究 使用共聚焦荧光显微镜和质谱分析。此外，翻译后 mDia 1和mDia 2的修饰，特别是乙酰化和磷酸化，以及这些修饰 影响HSC迁移的功能也将被表征。