Biomolecular Markers of Renal Allograft Status

描述（由申请人提供）：候选人：候选人，一名训练有素的临床医生，作为国际肾脏病学会的研究员加入导师的实验室。迄今为止，他已经确定丝氨酸蛋白酶-9（细胞毒性颗粒酶的内源性拮抗剂）的mRNA在肾同种异体移植物的急性排斥（AR）期间在尿中过度表达（Muthukumar T等人，Transplantation 2003），并且Foxp 3（调节性T细胞的特异性）的mRNA的尿细胞水平是肾同种异体移植物的AR的预后（Muthukumar T等人，N Engl J Med 2005）。他在纽约长老会-威尔康奈尔医学中心（NYPH-WCMC）完成了临床肾脏研究，并于2009年7月1日加入NYPH-WCMC担任全职教员。他在重新加入导师的实验室后开始了移植物内微小RNA表达模式的研究。工作环境：我们的机构，NYPH-WCMC倡导在我们的三机构网络内进行自由的科学探索，该网络由威尔康奈尔大学，纪念斯隆凯特琳癌症研究所和洛克菲勒大学组成。该环境的一些显著特征是：（1）导师的PCR核心研究实验室在定量PCR检测的开发/改进方面有着出色的记录，并作为PCR核心参与了NIH赞助的多中心临床试验，这些试验利用最先进的mRNA/miRNA分析设备进行器官移植;（ii）我们机构的跨学科肾移植计划;（iii）候选人的优秀共同导师包括F. Campagne（Bioinformatics，Weill Cornell），R. Ding（分子生物学，Weill Cornell），J.E. Schwartz（生物统计学，Weill Cornell/哥伦比亚），R.M. Steinman（Immunology，Rockefeller）和T. Tuschl（RNA Biology，Rockefeller）;（iv）拥有最先进技术的核心设施实验室;（v）提供出色生物信息学支持的计算生物医学研究所;（vi）由导师指导的临床免疫遗传学实验室;以及（vii）由NIH赞助的威尔康奈尔临床和转化科学中心。调研：候选人的近期目标：进一步发展研究技能，并通过结构化和逐步独立的研究和教学成分的培训成为一个独立的医生科学家。候选人的长期目标：提高移植领域的知识，帮助将基础研究成果转化为临床研究，优化器官移植。研究职业发展计划：我们的计划通过转化研究和教学活动的表现结合了研究培训。在获奖期间，候选人将致力于职业发展和培训，成为一名独立的医生科学家至少75%的努力。该研究还旨在使用生物学方法来解决差异基因表达模式的基础以及未来新疗法的开发。此外，候选人的共同导师是具有与候选人的新培训和职业发展高度相关的专业知识的杰出科学家，并将提供培训，正式审查他的进展，并与他的导师沿着促进他向独立的医生-科学家的过渡。候选人和他的导师将每周在一些场合进行正式和非正式的互动。候选人将有机会获得并将充分利用我们邻近的三机构网络中的可用资源，以进一步提高他的技能并获得知识。他已就读于K30临床与转化研究科学硕士，由威尔康奈尔大学临床与转化科学中心提供。研究项目：肾移植是治疗不可逆性肾衰竭的首选方法。多年来，短期结果有所改善，然而，近50%的患者在10年内失败。急性排斥反应（AR）和间质纤维化/肾小管萎缩（IF/TA）是同种异体移植物失败的重要因素。AR和IF/TA目前使用侵入性活检程序进行诊断。发展肾移植状态的非侵入性生物标志物是一个重要的目标。候选人将利用下一代测序技术，这是一项革命性的技术，可以发现AR和IFTA的移植物内生物标志物。然后，他将在研究的验证阶段使用在Mentor实验室改进的RT-PCR测定法，以产生mRNA/miRNA的绝对拷贝数。他将分析尿细胞和外周血细胞，以开发预测肾移植状态的非侵入性生物分子标记物。为了更好地理解机制并促进特定疗法的未来发展，他将建立模拟AR和IF/TA的体外细胞模型，并表征mRNA/miRNA谱，并研究短抑制性RNA对mRNA的调节。SA.1：通过表征分类为AR、IF/TA或正常的肾移植物活检组织的转录组，发现并验证AR和IF/TA的生物标志物。SA.2：建立体外细胞模型，在模拟AR或IF/TA的条件下表征转录组，并研究与AR和IF/TA相关的mRNA是否可以用siRNA调控。SA.3：通过尿细胞和外周血细胞的mRNA/miRNA分析，开发AR和IF/TA的非侵入性生物标志物。研究标本（同种异体移植物活检、尿液和血液）将来自入组两项NIH申办的研究的肾移植受者; R37 AI 051652和U 01 AI 08446。

DESCRIPTION (provided by applicant): Candidate: The candidate, a well trained clinician, joined the mentor's laboratory as a research fellow of the International Society of Nephrology. He has identified to date that mRNA for serine proteinase-9, an endogenous antagonist of cytotoxic granzyme, is over expressed in urine during acute rejection (AR) of renal allografts (Muthukumar T et al. Transplantation 2003), and that urinary cell levels of mRNA for Foxp3, a specification for regulatory T cells, are prognostic of AR of renal allografts (Muthukumar T et al. N Engl J Med 2005). He completed his clinical renal fellowship at the New York Presbyterian- Weill Cornell Medical Center (NYPH-WCMC) and joined as a full-time faculty member at NYPH-WCMC from July 1, 2009. He has initiated studies on intragraft micro RNA expression patterns upon re-joining the mentor's laboratory. Environment: Our institution, NYPH-WCMC advocates free scientific exploration within our tri-institutional network that is comprised of Weill Cornell, Memorial Sloan Kettering Cancer Institute and the Rockefeller University. Some of the significant features of the environment are: (1) the mentor's PCR core research laboratory with an excellent track record in the development/refinement of quantitative PCR assays and participation as the PCR Core in NIH sponsored multi-center clinical trials of organ transplantation utilizing state-of-the art equipment for mRNA/miRNA profiling; (ii) the interdisciplinary renal transplant program at our institution; (iii) the candidate's excellent co-mentors comprised of Drs. F. Campagne (Bioinformatics, Weill Cornell), R. Ding (Molecular Biology, Weill Cornell), J.E. Schwartz (Biostatistics, Weill Cornell/Columbia), R.M. Steinman (Immunology, Rockefeller) and T. Tuschl (RNA Biology, Rockefeller); (iv) Core facility laboratories with state-of-the art technologies; (v) an Institute of Computational Biomedicine providing outstanding bioinformatics support, (vi) the Clinical Immunogenetics Laboratory directed by the mentor; and (vii) the NIH sponsored Clinical and Translational Science Center at Weill Cornell. Research: Candidate's Immediate Goal: To further develop research skills, and become an independent physician-scientist through structured and progressively independent training with research and didactic components. Candidate's Long-Term Goal: To improve knowledge in the transplantation field and help translate basic research findings to the bedside and optimize organ transplantation. Research Career Development Plan: Our plan incorporates both research training via performance of translational research and didactic activities. During the award period, the candidate will commit a minimum of 75% effort towards career development and training towards becoming an independent physician scientist. The research proposed is also designed to use a biologic approach to address the basis for differential gene expression patterns as well development of novel therapeutics in the future. In addition, the candidate's co- mentors are outstanding scientists with expertise highly relevant to the candidate's new training and career progression, and will provide training, formally review his progress and facilitate, along with his mentor, his transition to an independent physician-scientist. The candidate and his mentor will interact both formally and informally in a number of settings on a weekly basis. The candidate will have access to and will take full advantage of the resources available in our neighboring tri-institutional network, to further enhance his skills and to acquire knowledge. He has enrolled in the K30 Master of Science in Clinical & Translational Investigation, offered by the Clinical & Translational Science Center at Weill-Cornell. Research Project: Renal transplantation is the preferred treatment for irreversible kidney failure. Short term outcomes have improved over the years, nevertheless, almost 50% fail by 10 years. Acute rejection (AR) and interstitial fibrosis/tubular atrophy (IF/TA) are significant contributors to allograft failure. AR and IF/TA are currently diagnosed using the invasive biopsy procedure. Development of noninvasive biomarkers of renal allograft status is an important goal. The candidate will take advantage of next-generation sequencing, a revolutionary technology to discover intragraft biomarkers of AR and IFTA. He will then use RT-PCR assays, improved in the Mentor's laboratory to yield absolute copy numbers of mRNAs/miRNAs, in the validation phase of the study. He will profile urinary cells and peripheral blood cells to develop noninvasive biomolecular markers predictive of renal allograft status. To better understand mechanisms and facilitate future development of specific therapies, he will establish in-vitro cellular models mimicking AR and IF/TA and characterize mRNA/miRNA profiles and investigate the regulation of mRNAs by short inhibitory RNAs. SA.1: To discover and validate biomarkers of AR and IF/TA by characterizing the transcriptome of renal allograft biopsies classified as AR, IF/TA or normal. SA.2: To establish in-vitro cellular models and characterize transcriptomes under conditions mimicking AR or IF/TA and investigate whether mRNAs associated with AR and IF/TA can be regulated with siRNAs. SA.3: To develop noninvasive biomarkers of AR and IF/TA by mRNA/miRNA profiling of urinary cells and peripheral blood cells. Study specimens (allograft biopsies, urine and blood) will be from kidney transplant recipients enrolled in two-NIH-sponsored studies; R37 AI051652 and U01 AI08446.