Noninvasive Biomarkers of Microcirculatory Injury in the Human Kidney Allograft

人肾同种异体移植物微循环损伤的非侵入性生物标志物

• 批准号: 9206157
• 负责人: Thangamani Muthukumar
• 金额: $ 8.48万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9206157
• 关键词: Acute; Allografting; Antibodies; Award; Biological Assay; Biological Markers; Biopsy; Blood; Blood Tests; Blood Vessels; Blood capillaries; Cells; Clinic; Clinical; Cohort Studies; Core Biopsy; Data Analyses; Development; Diagnosis; Diagnostic; Discriminant Analysis; Endothelium; Enrollment; Ensure; Failure; Genes; Histologic; Histology; Hospitals; Human; Individual; Inflammation; Injury; Investigation; Kidney Transplantation; Laboratories; Measures; Mediating; Medical; Mentors; MicroRNAs; Modeling; Needle biopsy procedure; Patients; Physicians; Positioning Attribute; Procedures; Process; RNA; Reading; Research; Research Personnel; Research Project Grants; Sampling; Sampling Errors; Serum; Small RNA; Standardization; Statistical Models; Study of serum; System; Techniques; Testing; Time; Translations; Transplant Recipients; Transplantation; Tubular formation; United States National Institutes of Health; Validation; Work; base; cDNA Library; capillary; career; circulating microRNA; cohort; cost; deep sequencing; experimental study; extracellular; innovation; kidney allograft; laboratory curriculum; liver allograft; new technology; noninvasive diagnosis; novel; programs; public health relevance; response; tool; transcriptome

 DESCRIPTION (provided by applicant): Kidney transplants fail over time. Injury due to antibodies that target cells of the blood vessels within the transplant kidney (microcirculatory endothelium) is a major reason why the transplants fail. Clinical tools to predict this injury are not available. MicroRNAs in the blood could serve as markers of injury. We propose to develop microRNA (miRNA)-based blood tests for the diagnosis of antibody-mediated microcirculatory inflammation/injury. We have carefully selected a group of 180 kidney transplant recipients at our hospital who had serum samples collected and stored in our laboratory at the time of undergoing an allograft biopsy. The AMI group (n=60) consists of 60 serum samples from 60 kidney transplant recipients with biopsy diagnosis of antibody-mediated microcirculatory inflammation. The No AMI group (n=120) consists of 120 serum samples from 120 kidney transplant recipients whose biopsy did not have such inflammation. This group consists of patients with biopsy diagnosis of acute cellular rejection, acute tubular injury, and normal allograft biopsy (n=40 each). The Specific Aims are to: (1) Identify circulating extracellular miRNAs in serum of kidney graft recipients that are diagnostic of AMI in the kidney allograft, (2) Develop a statistical model (signature) diagnostic of AMI in the kidney allograft, (3) Validate the diagnostic model in serum samples obtained from an independent external cohort of kidney transplant recipients. We will use high throughput barcoded deep sequencing to characterize the serum miRNA transcriptome. We will use miRNA-specific qPCR assay to accurately quantify selected individual miRNAs, for their quick translation to the clinic. We will compare the serum miRNA transcriptome from the two groups (AMI vs. No AMI) and identify miRNAs that are different between the two. We will then develop PCR assays to accurately quantify the selected miRNAs. We will develop statistical models consisting of combination of miRNAs that best predict the diagnosis. We will statistically determine the clinical benefit of using such a model fr diagnosing AMI. To ensure that the statistical model works in different groups of patients, we will do PCR assays and test the statistical model (signature) in another group of 105 kidney transplant recipients (35 AMI and 70 No AMI) from a different hospital.

 描述(由申请人提供)：肾移植随着时间的推移而失败。针对移植肾内血管细胞(微循环内皮细胞)的抗体造成的损伤是移植失败的主要原因。目前还没有预测这种损伤的临床工具。血液中的microRNAs可以作为损伤的标志。我们建议开发基于microRNA(MiRNA)的血液检测来诊断抗体介导的微循环炎症/损伤。我们精心挑选了一组180例肾移植受者，他们在接受同种异体肾移植活检时收集并储存了血清样本。急性心肌梗死组(n=60)包括来自60例肾移植受者的60份血清样本，活检诊断为抗体介导的微循环炎症。非急性心肌梗死组(n=120)包括来自120名肾移植受者的120份血清样本，他们的活检组织中没有这种炎症。这组患者包括活检诊断为急性细胞排斥反应、急性肾小管损伤和正常同种异体移植肾活检的患者(n=40)。其具体目的是：(1)确定肾移植受者血清中循环细胞外miRNAs对移植肾急性心肌梗死的诊断；(2)建立诊断移植肾急性心肌梗死的统计模型(SIGN)；(3)验证 从独立的肾移植受者外部队列中获得的血清样本的诊断模型。我们将使用高通量条形码深度测序来表征血清miRNA转录组。我们将使用miRNA特异的qPCR方法来准确地定量选定的单个miRNAs，以便将它们快速翻译到临床上。我们将比较两组患者(急性心肌梗死和非急性心肌梗死)的血清miRNA转录组，并确定两者之间的不同miRNAs。然后，我们将开发聚合酶链式反应分析，以准确地定量选定的miRNAs。我们将开发由miRNAs组合组成的统计模型，以最好地预测诊断。我们将从统计学上确定使用这种模型诊断急性心肌梗死的临床益处。为确保统计模式适用于不同组别的病人，我们会 对另一组来自不同医院的105例肾移植受者(35例急性心肌梗死和70例非急性心肌梗死)进行聚合酶链式反应分析并检验统计模型(Signature)。