Adrenal Regeneration: Stem Cells and Lineage Development

描述(由申请人提供)： 许多疾病状态是由组织的丢失或破坏引起的，例如，胰岛细胞丢失导致I型糖尿病，肾上腺皮质细胞丢失导致艾迪森氏病。这些疾病的最终治疗将包括(1)逆转破坏过程和(2)恢复正常组织。组织再生是将丢失或受损的组织恢复到其正常状态的过程。调节这一动态和基本过程的机制仍然不完全清楚。两个主要机制包括调节(1)组织特异性成体干细胞和(2)谱系发育，这可能涉及细胞分化、去分化和转分化。 利用一个经典的组织再生模型--小鼠肾上腺皮质，我们将研究这些基本过程。使用荧光激活细胞分选，我们将从经历各种受控操作的动物中鉴定、分离和鉴定肾上腺前体细胞。然后，将在移植模型系统中检查祖细胞的分化潜力。利用基因打靶方法和Cre-loxP技术，我们将对小鼠肾上腺细胞进行基因修饰，以便进行谱系追踪和组织特异性基因敲除实验。这些实验旨在阐明肾上腺发育和分区的基本机制，并开始定义肾上腺再生的分子途径。 这项工作是在高度支持的环境中进行的，建议研究的每个主要领域都有专门知识。此外，这项工作建立在PI先前的经验基础上，通过结合基因打靶和Cre-loxP技术的现代工具，产生转基因动物来研究基因表达和谱系发育。最后，通过将这项工作与作为儿科内分泌科医生的有限临床责任(10%的时间)相结合，PI将继续从临床接触中受益，这往往会导致对疾病机制的新见解。预计这项工作将使和平研究所能够过渡到独立研究人员的职业生涯。

DESCRIPTION (provided by applicant): A number of disease states arise from the loss or destruction of tissue, e.g., loss of pancreatic islets cells results in Type I Diabetes and loss of adrenal cortical cells results in Addison's Disease. The ultimate treatment of these conditions would involve (1) reversal of the destructive process and (2) restoration of normal tissue. Tissue regeneration is the process by which lost or damaged tissue is restored to its normal state. The mechanisms, which regulate this dynamic and essential process, remain incompletely understood. Two major mechanisms include the regulation of (1) tissue-specific adult stem cells and (2) lineage development, which likely involves cellular differentiation, de-differentiation and trans-differentiation. Employing a classic model of tissue regeneration, the mouse adrenal cortex, we will investigate these fundamental processes. Using fluorescence activate cell sorting, we will identify, isolate and characterize adrenal progenitor cells from animals undergoing a variety of controlled manipulations. Progenitor cells will then be examined for their differentiation potential in transplantation model systems. Using a gene targeting approach and Cre-LoxP technology we will genetically modify mouse adrenal cells in order to perform lineage tracing and tissue-specific knockout experiments. These experiments are designed to elucidate fundamental mechanisms of adrenal development and zonation and begin to define the molecular pathway of adrenal regeneration. This work is being performed in a highly supportive environment with expertise in each major area proposed for study. Furthermore, this work builds upon the prior experiences of the PI, generating transgenic animals to study gene expression and lineage development, by incorporating the modern tools of gene targeting and Cre-LoxP technology. Finally, by combining this work with limited clinical responsibility (10% time) as a Pediatric Endocrinologist, the PI will continue to benefit from clinical encounters, which often lead to novel insights into the mechanisms of disease. It is expected that this work will be enable the PI to transition to a career as an independent researcher.