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MD/PhD博士后科学家，在Albert的分子和细胞生物学方面进行了广泛的培训 爱因斯坦医学院和怀特海研究所。我的研究重点是MDIA FORMIN的角色 造血细胞上的蛋白质。 MDIA formin蛋白在许多人的迁移中具有良好的作用 不同的细胞类型。但是，它们对造血细胞迁移的影响，尤其是造血细胞的迁移 干细胞（HSC），知之甚少。与怀特黑德研究所的理想研究环境和 Harvey Lodish博士的出色指导，我计划通过 在血肿干细胞和细胞迁移的领域发展专业知识，这对我的长期至关重要 成为成功的独立调查员的职业目标。 我提出的研究的目的是表征MDIA formins的功能作用，特别是MDIA1 和MDIA2，在造血干细胞（HSC）植入和迁移上。 HSC植入和迁移是 对于成功的骨髓移植至关重要，这已成为治疗的常规临床策略 患有许多血液相关疾病的患者，例如性贫血，法科尼贫血，镰状细胞贫血，β Thalassyia专业，白血病，淋巴瘤，多发性骨髓瘤和许多免疫缺陷障碍。这 调节HSC植入和迁移的细胞内信号通路通常直接或间接靶向 通过调节肌动蛋白聚合的蛋白质的肌动蛋白细胞骨架网络。 MDIA formin蛋白是一个主要的肌动蛋白核蛋白，其在HSC中的功能知之甚少。到 阐明MDIA formins，特别是MDIA1和MDIA2在HSC迁移中的作用 将使用小鼠模型和分子细胞生物学技术。具体而言，鼠标模型不足 将生成MDIA1或MDIA2。使用这些小鼠，骨髓和胎儿肝lin-sca1+ kit+（LSK）细胞， 富集的HSC种群将被纯化以进行体外迁移测定，短期归住和长期 术语植入分析。与Drs合作。 Tzutzuy Ramirez Hernandez和Maki Murata-Hori 新加坡的Temasek Life Sciences实验室，MDIA1和MDIA2与肌动蛋白的动力学相互作用 将研究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 使用共聚焦荧光显微镜和质谱分析。另外，翻译后 MDIA1和MDIA2的修饰，尤其是乙酰化和磷酸化，以及这些修饰如何 影响他们在HSC迁移中的功能也将被表征。