Analysis of mDia formins in hematopoietic stem cell engraftment and migration

我是一名医学博士/博士后科学家，在阿尔伯特大学接受过分子和细胞生物学方面的广泛培训 爱因斯坦医学院和怀特黑德研究所。我的研究重点是媒体形成的作用 造血细胞上的蛋白质。Mdia Forin蛋白在许多细胞的迁移过程中具有公认的作用。 不同的细胞类型。然而，它们对造血细胞的迁移，特别是对造血细胞的迁移的影响 干细胞(HSCs)，人们对此知之甚少。怀特黑德研究所理想的研究环境和 哈维·洛迪什博士的出色指导，我计划通过以下方式扩大我现有的科学和智力技能 在造血干细胞和细胞迁移领域发展专业知识，这对我的长期职业生涯至关重要 职业目标是成为一名成功的独立调查员。 我提议的研究的目标是表征mdia福尔马林，特别是mdia1的功能作用。 和mDia2，对造血干细胞(HSC)的植入和迁移。HSC的植入和迁移是 对于成功的骨髓移植至关重要，这已经成为一种常规的临床治疗策略 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、贝塔 地中海贫血、白血病、淋巴瘤、多发性骨髓瘤等多种免疫缺陷疾病。这个 调控HSC植入和迁移的细胞内信号通路往往直接或间接靶向于 肌动蛋白通过调节肌动蛋白聚合的蛋白质形成细胞骨架网络。媒体媒体 Forin蛋白是一个主要的肌动蛋白核家族，对其在HSCs中的功能知之甚少。至 阐明mdia Forins，特别是mDia1和mDia2在转基因HSC迁移中的作用 将使用小鼠模型和分子细胞生物学技术。具体地说，小鼠模型缺乏 将生成mDia1或mDia2。用这些小鼠、骨髓和胎肝LIN-Sca1+Kit+(LSK)细胞，以及 浓缩的HSC群体，将被纯化以进行体外迁移试验、短期归巢和长期归巢 术语嫁接分析。与The Tzutzuy Ramirez Hernandez博士和Maki Murata-Hori博士合作 新加坡淡马锡生命科学实验室，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的迁移。 采用共聚焦荧光显微镜和质谱仪进行分析。此外，翻译后 MDia1和mDia2的修饰，特别是乙酰化和磷酸化，以及这些修饰是如何 还将对其在HSC迁移中的作用进行描述。

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.