Biomolecular Markers of Renal Allograft Status

同种异体肾移植状态的生物分子标志物

• 批准号: 8372404
• 负责人: Thangamani Muthukumar
• 金额: $ 13.11万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8372404
• 关键词: Acute; Address; Advisory Committees; Advocate; Allogenic; Allografting; Atrophic; Award; Basic Science; Bioinformatics; Biological Assay; Biological Markers; Biological Process; Biology; Biometry; Biopsy; Blood; Blood Cells; Caring; Cell model; Cells; Center for Translational Science Activities; Chronic; Clinical; Clinical Sciences; Commit; Core Facility; Data; Dendritic Cells; Development; Development Plans; Diagnosis; Diagnostic; Diagnostic tests; Enrollment; Environment; Epithelial Cells; Equipment; Faculty; Failure; Fellowship; Fibrosis; Future; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Genomics; Goals; Graft Rejection; Granzyme; Head; Heart Diseases; Hemorrhage; Human; Immune; Immunogenetics; Immunology; In Vitro; Institutes; Institution; International; Investigation; Kidney; Kidney Failure; Kidney Transplantation; Knowledge; Laboratories; Laboratory Research; Lead; Letters; Literature; Malignant Neoplasms; Master of Science; Medical center; Mentors; Messenger RNA; MicroRNAs; Modeling; Molecular Biology; Molecular Profiling; Multi-Institutional Clinical Trial; Nephrology; New York; Organ Transplantation; Outcome; Patients; Pattern; Performance; Phase; Physicians; Presbyterian Church; Procedures; Protocols documentation; RNA; RNA Sequence Analysis; RNA Sequences; Regulation; Regulator Genes; Regulatory T-Lymphocyte; Research; Research Project Grants; Research Training; Resources; Reverse Transcriptase Polymerase Chain Reaction; Scientist; Serine Protease; Societies; Specimen; Structure; T-Lymphocyte; Technology; Testing; Time; Tissue-Specific Gene Expression; Training; Translating; Translational Research; Transplant Recipients; Transplantation; Tubular formation; United States National Institutes of Health; Universities; Urine; Validation; base; career; career development; cohort; cytotoxic; design; diagnostic accuracy; improved; in vitro Model; insight; interstitial; kidney allograft; knock-down; member; model development; next generation; next generation sequencing; novel therapeutics; prognostic; programs; public health relevance; skills; urinary

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.

描述（由申请人提供）： 候选人： 候选人是一位训练有素的临床医生，加入导师的实验室，担任国际肾脏病学会的研究员。迄今为止，他已确定丝氨酸蛋白酶 9（一种细胞毒性颗粒酶的内源性拮抗剂）的 mRNA 在同种肾移植物的急性排斥 (AR) 期间在尿液中过度表达（Muthukumar T 等人，移植 2003 年），并且尿细胞中 Foxp3（调节性 T 细胞的规范）的 mRNA 水平是肾同种异体移植物 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) 完成了临床肾脏奖学金，并于 2009 年 7 月 1 日起成为 NYPH-WCMC 的全职教员。重新加入导师实验室后，他开始了移植物内 micro RNA 表达模式的研究。环境：我们的机构 NYPH-WCMC 提倡在由威尔康奈尔大学、纪念斯隆凯特琳癌症研究所和洛克菲勒大学组成的三机构网络内进行自由科学探索。该环境的一些显着特征包括：(1) 导师的 PCR 核心研究实验室在定量 PCR 检测的开发/改进方面拥有出色的记录，并作为 PCR 核心参与 NIH 赞助的器官移植多中心临床试验，利用最先进的 mRNA/miRNA 分析设备； (ii) 我们机构的跨学科肾移植项目； (iii) 候选人的优秀共同导师包括博士。 F. Campagne（生物信息学，威尔康奈尔）、R. Ding（分子生物学，威尔康奈尔）、J.E. Schwartz（生物统计学，威尔康奈尔/哥伦比亚）、R.M. Steinman（免疫学，洛克菲勒）和 T. Tuschl（RNA 生物学，洛克菲勒）； (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) 导师指导的临床免疫遗传学实验室； (vii) 美国国立卫生研究院 (NIH) 赞助的威尔康奈尔临床和转化科学中心。研究：候选人的近期目标：进一步发展研究技能，并通过包含研究和教学内容的结构化且逐步独立的培训成为一名独立的医师科学家。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并优化器官移植。研究职业发展计划：我们的计划包括通过转化研究和教学活动进行的研究培训。在奖励期间，候选人将至少投入 75% 的努力用于职业发展和培训，以成为一名独立的医师科学家。拟议的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，候选人的导师都是杰出的科学家，他们的专业知识与候选人的新培训和职业发展高度相关，并将提供培训，正式审查他的进展，并与他的导师一起促进他向独立医师科学家的过渡。候选人和他的导师每周将在多种场合进行正式和非正式的互动。候选人将能够访问并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取知识。他已报名参加由威尔康奈尔大学临床与转化科学中心提供的 K30 临床与转化研究理学硕士课程。研究项目：肾移植是不可逆性肾衰竭的首选治疗方法。多年来，短期结果有所改善，但近 50% 的结果在 10 年后会失败。急性排斥反应（AR）和间质纤维化/肾小管萎缩（IF/TA）是同种异体移植失败的重要原因。 AR 和 IF/TA 目前使用侵入性活检程序进行诊断。开发肾同种异体移植状态的非侵入性生物标志物是一个重要目标。候选人将利用下一代测序这一革命性技术来发现 AR 和 IFTA 的移植内生物标志物。然后，他将在研究的验证阶段使用在 Mentor 实验室改进的 RT-PCR 测定法来产生 mRNA/miRNA 的绝对拷贝数。他将分析尿细胞和外周血细胞，以开发预测肾同种异体移植状态的非侵入性生物分子标记。为了更好地理解机制并促进特定疗法的未来开发，他将建立模拟 AR 和 IF/TA 的体外细胞模型，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 或正常的肾同种异体移植物活检的转录组来发现和验证 AR 和 IF/TA 的生物标志物。 SA.2：建立体外细胞模型并在模拟 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 进行调节。 SA.3：通过尿细胞和外周血细胞的 mRNA/miRNA 分析来开发 AR 和 IF/TA 的非侵入性生物标志物。研究样本（同种异体移植物活检、尿液和血液）将来自参加两项 NIH 赞助研究的肾移植受者； R37 AI051652 和 U01 AI08446。