Cell Cycle Related Transdifferentation Plasticity

细胞周期相关的转分化可塑性

• 批准号: 7084635
• 负责人: MEHRDAD ABEDI
• 金额: $ 12.13万
• 依托单位: ROGER WILLIAMS HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-14 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7084635
• 关键词: biological models; blood fractionation; bone marrow; bone marrow transplantation; cell adhesion; cell cycle; cell differentiation; cell migration; cell type; cytokine; flow cytometry; fluorescence microscopy; fluorescent in situ hybridization; genetically modified animals; green fluorescent proteins; heart; hematopoietic tissue; histogenesis; laboratory mouse; liver disorder; musculoskeletal injury; stem cells; striated muscles

DESCRIPTION (provided by applicant): Mehrdad Abedi MD is a HematologylOncology postdoctorate fellow and a candidate for a K08 grant under the mentorship of Dr. Peter J. Quesenberry. The applicant wishes to pursue an academic career with his research focus in the field of stem cell biology. Marrow derived cells have the capacity to regenerate many different cell types in their appropriate tissues or in vitro. Functional stem cell plasticity connected with different phases of cell cycle and the transdifferentation ability of these cells have not been well charaterized. With cytokine driven cell cycle transit, adhesion protein expression, engraftment and differentiation fluctuates reversibly and these fluctuations appear to be tied to specific phases of cell cycle. The hypothesis is that the transdifferentation plasticity of marrow will also fluctuate with phase of cell cycle and that the critical initial event in this process relates to homing into different tissues. Besides giving dramatic new insights into stem cell biology, this project offers unique potential for various clinical tissue restorative approaches. We plan to study different marrow fractions (whole marrow, lin-sca+, rhodamine low Hoescht low, GFP+ Hoescht low side population, and clonally replaced marrow) through all points in a cytokine induced cell cycle tansit. We will determine the homing ability, short and long-term fate of these cells that go to heart, skeletal muscle and liver in untreated or organ injured mice and their ability to transdifferentiate in the above tissues. Two murine transplant models that allow for cell tracking will be used; the BALB/c male to female model and the B6 GFP transgenic model. Donor cells will be identified using FISH for the Y-chromosome and fluorescence or Ab staining for GFP. Immediate homing will be done marking the cells with a cytoplasmic dye. Double labeling approaches on frozen or paraffin sections will be carried out to identify donor origin and cell type. Mice will also be repopulated with "marked" marrow fractions and then hematopoietic mobilization will be done with or without heart, skeletal muscle or liver injury and again look at short and long-term cell fate and transdifferentation. High-speed cell sorting, fluorescent microscopy imaging and large event FACS analysis will evaluate short term homing. This project will explore the functional plasticity of marrow to transdifferentate into nonhematopoietic tissues (heart, skeletal muscle and in liver).

描述（由申请人提供）： Mehrdad Abedi 医学博士是血液学和肿瘤学博士后研究员，也是 Peter J. Quesenberry 博士指导下 K08 资助的候选人。申请人希望从事学术职业，其研究重点是干细胞生物学领域。骨髓来源的细胞具有在适当的组织或体外再生许多不同细胞类型的能力。与细胞周期不同阶段相关的功能性干细胞可塑性以及这些细胞的转分化能力尚未得到很好的表征。随着细胞因子驱动的细胞周期转变，粘附蛋白表达、植入和分化可逆地波动，并且这些波动似乎与细胞周期的特定阶段相关。该假设是，骨髓的转分化可塑性也会随着细胞周期的阶段而波动，并且该过程中的关键初始事件与归巢到不同组织有关。除了为干细胞生物学提供引人注目的新见解外，该项目还为各种临床组织修复方法提供了独特的潜力。我们计划通过细胞因子诱导的细胞周期转变中的所有点来研究不同的骨髓组分（全骨髓、lin-sca+、罗丹明低 Hoescht 低、GFP+ Hoescht 低侧群体和克隆替代骨髓）。我们将确定这些细胞在未经治疗或器官损伤的小鼠中进入心脏、骨骼肌和肝脏的归巢能力、短期和长期命运，以及它们在上述组织中转分化的能力。将使用两种允许细胞追踪的小鼠移植模型； BALB/c 男变女模型和 B6 GFP 转基因模型。将使用 FISH 检测 Y 染色体，使用荧光或 Ab 染色检测 GFP 来鉴定供体细胞。立即归巢将用细胞质染料标记细胞。将在冷冻或石蜡切片上进行双标记方法，以确定供体来源和细胞类型。小鼠还将被重新植入“标记的”骨髓部分，然后在有或没有心脏、骨骼肌或肝脏损伤的情况下进行造血动员，并再次观察短期和长期的细胞命运和转分化。高速细胞分选、荧光显微镜成像和大事件 FACS 分析将评估短期归巢。该项目将探索骨髓转分化为非造血组织（心脏、骨骼肌和肝脏）的功能可塑性。