CYCLOPHILIN C FUNCTION

描述 (改编自申请人的摘要)亲环素C(CYP C)是一个 高度保守的环孢素A(CsA)结合蛋白家族 存在于细菌、单细胞生物体和所有动植物物种中 测试过。这些普遍存在的蛋白质，占总蛋白质的0.1%到0.4% 细胞蛋白质仍然是个谜。利用CYP C进行的研究是第一次 确定钙调神经磷酸酶(CN)为免疫抑制剂CsA的靶标，以及 与FKBP-12的类似研究迅速表明，CN是一种常见的 CsA和FK506的下游目标。有了这些结果，关键部分 在他的博士工作中，他在1992年不情愿地放下了移液管，完成了 他接受了医学博士培训，并于1993年开始儿科实习和住院医生工作 波士顿的儿童医院。在他招生的时候，其他人 正在提交意见书--在他之后进行了大量的工作 关于CN在T细胞活化中作用的初步观察。不过， 回到实验室后，作为儿科血红素/ONC的研究员，它已经成为 显然，关于CYP C的功能仍有许多需要了解的地方(和 在没有外源药物的情况下)。实验 目的是阐明小鼠细胞色素C的功能，并由此推断， 概述了这些古老而普遍存在的蛋白质的功能。这个 具体目标包括鉴定小鼠的细胞色素C基因； 组织特异性表达启动子/增强子元件的分析 (骨髓基质，皮质髓质交界处的肾小管细胞， 卵巢基质细胞和间质细胞)和IL-1的诱导 含绿色荧光的CYP C-/-小鼠的产生 CYP C基因座控制下的蛋白质转基因及分析 细胞色素C细胞内定位的决定因素。单元格的GFP标记 一个活性的CYP C启动子(CYP C+/-和-/-细胞)将促进 直接测试细胞色素C生物功能的实验(如果-/-动物是 可行)。骨髓基质的绿色荧光蛋白标记将允许 基质细胞前体细胞在发育和定向发育过程中的鉴定 测试这类细胞在移植模型中的重要性。这个 达纳·法伯酒店的设施和员工，与富人相结合 靠近哈佛和麻省理工学院所提供的智力资源，提供了 开展这一研究计划的理想环境是 旨在推进弗里德曼博士的培训，整合他在 血液学和亲环素生物学，并促进他向 未来成为一名独立的调查员。(摘要结束)

DESCRIPTION (Adapted from applicant's abstract) Cyclophilin C (cyp C) is a member of a highly conserved and growing family of cyclosporine A (CsA) binding proteins found in bacteria, unicellular organisms and all plant and animal species tested. These ubiquitous proteins, which represent 0.1% to 0.4% of total cellular protein remain enigmatic. Studies utilizing cyp C were the first to identify calcineurin (CN) as the target of the immunosuppressant CsA, and analogous studies with FKBP-12 rapidly demonstrated that CN was a common downstream target of both CsA and FK506. With these results, key portions of his Ph.D. work, he reluctantly put down his pipette in 1992, completed his M.D. training and in 1993 began Pediatric Internship and Residency at The Children's Hospital in Boston. While he was doing admissions, others were making submissions--a tremendous amount of work has followed his initial observations regarding the role of CN in T cell activation. Still, upon returning to the lab as a fellow in Pediatric Heme/Onc it has become clear that much remains to be learned regarding the function of cyp C (and cyclophilins in general) in the absence of exogenous drugs. Experiments aimed at elucidating the function of murine cyp C, and, by inference, the function of these ancient and ubiquitous proteins are outlined. The specific aims include the characterization of the murine cyp C gene; an analysis of promoter/enhancer elements leading to tissue specific expression (bone marrow stroma, renal tubular cells at the corticomedullary junction, ovarian stromal cell and tesucular interstitial cells) and IL-1 induction of cyp C; the generation of cyp C -/- mice containing a green fluorescent protein transgene under control of the cyp C locus; and analysis of determinants of cyp C intracellular localization. GFP marking of cells with an active cyp C promoter (both cyp C +/- and -/- cells) will facilitate experiments to directly test biologic functions of cyp C (if -/- animals are viable). GFP marking of bone marrow stroma will allow both the identification of stromal cell precursors during development and direct testing of the importance of such cells in transplantation models. The facilities and staff at the Dana Farber, in combination with the rich intellectual resources afforded by proximity to Harvard and MIT, provide an ideal environment for carrying out this program of research which is designed to advance Dr. Friedman's training, integrate his interests in hematology and cyclophilin biology, and facilitate his transition towards a future as an independent investigator. (End of Abstract)