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: 8585052
• 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/8585052
• 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.

应聘者：应聘者是一名训练有素的临床医生，曾作为国际肾脏病学会研究员加入导师实验室。到目前为止，他已经发现细胞毒性颗粒酶的内源性拮抗剂丝氨酸蛋白酶-9的mRNA在肾移植急性排斥(AR)期间在尿中过度表达(Muthukumar T等人)。Foxp3的尿细胞信使核糖核酸水平是肾移植后AR的预测指标(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)完成了他的临床肾脏研究员资格，并于2009年7月1日加入纽约长老会-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。环境：我们的机构NYPH-WCMC倡导在我们由威尔·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个机构网络内进行自由的科学探索。环境的一些重要特征是：(1)Mentor的聚合酶链式反应核心研究实验室在开发/改进定量聚合酶链式反应分析方面有着良好的记录，并作为聚合酶链式反应核心参与了NIH赞助的多中心器官移植临床试验，利用最先进的mRNA/miRNA分析设备；(Ii)我们机构的跨学科肾移植计划；(Iii)候选人的优秀合作导师包括F.Campagne博士(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)提供杰出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)NIH赞助的魏尔康奈尔临床和转化科学中心。研究：应聘者的近期目标：进一步发展研究技能，并通过研究和教学内容的结构化和渐进式独立培训成为一名独立的内科医生-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床边研究成果，并优化器官移植。研究职业发展计划：我们的计划包括通过翻译研究的表现进行研究培训和教学活动。在获奖期间，候选人将在职业发展和培训方面做出至少75%的努力，以成为一名独立的内科科学家。提出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，候选人的共同导师是杰出的科学家，具有与候选人的新培训和职业发展高度相关的专业知识，他们将提供培训，正式审查他的进展，并与他的导师一起促进他向独立的内科科学家过渡。候选人和他的导师每周都会在许多场合进行正式和非正式的互动。候选人将能够使用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和获取知识。他已经报名参加了由威尔康奈尔大学临床与翻译科学中心提供的临床与翻译调查K30理学硕士课程。研究项目：肾移植是不可逆性肾功能衰竭的首选治疗方法。多年来，短期结果有所改善，然而，近50%的人在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)是同种异体肾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，这是一项革命性的技术，可以发现AR和IFTA的移植物内生物标记物。然后，在研究的验证阶段，他将使用在导师的实验室改进的RT-PCR分析来产生mRNAs/miRNAs的绝对拷贝数。他将分析尿液细胞和外周血细胞，以开发预测肾移植状态的非侵入性生物分子标记物。为了更好地了解机制并促进未来特定疗法的发展，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RNA对mRNAs的调节。SA.1：通过鉴定AR、IF/TA或正常肾活检组织的转录组，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，在模拟AR或IF/TA的条件下鉴定转录本，并研究与AR和IF/TA相关的mRNAs是否受siRNAs的调控。SA.3：通过尿液细胞和外周血细胞的mRNA/miRNA图谱，开发AR和IF/TA的非侵入性生物标志物。研究样本(同种异体移植活检、尿液和血液)将来自NIH赞助的两项研究中登记的肾移植受者；R37 AI051652和U01 AI08446。