Mechanisms of Asymmetric Cell Division in Developmental Hematopoiesis

项目摘要/摘要 所有血细胞类型都来自单一的造血干细胞(HSC)前体。HSC必须 通过自我更新的对称细胞分裂和子代细胞的分化来平衡扩张 不对称细胞分裂(ACD)以产生功能正常的血液系统。当ACD被扰乱时，深层血液 疾病通常以特定血细胞类型的不成比例产生或不受控制为特征 血祖细胞的扩增。拟议项目的目标是了解ACD如何影响 发育中的造血和血细胞命运的选择。目标1将通过以下方式描述ACD的发生方式 利用基于RNAi的耗竭在造血祖细胞和 免疫荧光技术。目标2将确定ACD如何影响动态平衡期间的世系选择 和应激通过监测ACD在动态平衡期间和应激源之后的血细胞命运 比如缺氧或受伤。最终，这些实验将有助于阐明一种空间和机制 ACD在造血中的模型建立有助于加速进展的果蝇模型 在哺乳动物ACD中，造血起作用。 我的职业目标是成为一家顶级学术机构的一名成功的独立研究员。我 目的是领导一项研究计划，调查来自利基和微环境的外部线索 影响细胞极性和细胞分裂，从而影响HSC的自我更新和血细胞谱系选择。要实现 这些目标，我将利用基础细胞生物学和细胞骨架研究方面的技术和培训相结合 血液学和先进成像技术的遗传解剖。我将继续接受拟议中的培训 在Utpal Banerjee博士的指导下，他拥有丰富的使用基因技术进行解剖的经验 以果蝇为模型系统研究造血的分子基础。另外，我的建议是 委员会将提供关键的支持和资源，帮助我将我的发现转化为哺乳动物系统和 开发尖端成像技术，在动态平衡期间实时监测血液系统 压力。我将在国内和国际会议上展示我的工作并传播我的发现，以帮助 推进我的研究，建立合作关系，并在造血领域站稳脚跟。这个项目将 通过帮助我确定使用的可行性，帮助我过渡到独立研究职位 为研究果蝇在造血过程中的ACD和前期工作奠定了基础。加州大学洛杉矶分校是 这是从事这项工作的绝佳场所，因为它提供了丰富的资源、研究合作和 职业发展机会帮助我提升我的事业。作为一名初级教员，我将参加 培训活动的重点是领导技能、赠款撰写、在影响力较大的期刊上发表文章、实验室 管理和指导，帮助我培养管理一个成功的研究团队所需的技能。

Project Summary/Abstract All blood cell types are derived from a single hematopoietic stem cell (HSC) precursor. HSCs must balance expansion through self-renewing symmetric cell division with differentiation of progeny cells through asymmetric cell division (ACD) to produce a functional blood system. When ACD is disrupted, profound blood disorders occur often characterized by disproportionate production of specific blood cell types or uncontrolled expansion of blood progenitors. The goal of the proposed project is to understand how ACD influences developmental hematopoiesis and blood cell fate choice. Aim 1 will characterize how ACD occurs by identifying molecular players using RNAi-based depletion in hematopoietic progenitors and immunofluorescence techniques. Aim 2 will determine how ACD influences lineage choice during homeostasis and stress by monitoring blood cell fate upon disruption of ACD during homeostasis and after stressors such as oxygen deprivation or injury. Ultimately, these experiments will help elucidate a spatial and mechanistic model for ACD in hematopoiesis while establishing a Drosophila model that will help accelerate advancements in mammalian ACD hematopoiesis work. My career goal is to become a successful independent researcher at a top-tier academic institution. I aim to lead a research program that investigates how extrinsic cues from the niche and microenvironment influence cell polarity and cell division to impact HSC self-renewal and blood cell lineage choice. To achieve these goals, I will utilize techniques and training in basic cell biology and cytoskeleton research combined with genetic dissection of hematopoiesis and advanced imaging techniques. I will pursue the proposed training under the mentorship of Dr. Utpal Banerjee who has extensive experience using genetic techniques to dissect the molecular underpinnings of hematopoiesis using Drosophila as a model system. Additionally, my advisory committee will provide key support and resources to help me translate my findings to mammalian systems and develop cutting edge imaging techniques to monitor the blood system in real-time during homeostasis and stress. I will present my work and disseminate my findings at national and international conferences to help advance my research, build collaborations, and establish myself in the hematopoiesis field. This project will facilitate my transition to an independent research position by helping me establish the feasibility of using Drosophila to study ACD during hematopoiesis and a foundation of preliminary work to build upon. UCLA is an excellent place to pursue this work as it provides a rich landscape of resources, research collaborations, and professional development opportunities to help me advance my career. As a junior faculty, I will take part in training activities focused on leadership skills, grant writing, publishing in high impact journals, laboratory management, and mentoring to help me develop the skills needed to run a successful research team.