Clinical utility of extracellular RNA as marker of kidney disease progression

细胞外 RNA 作为肾脏疾病进展标志物的临床应用

• 批准号: 9128779
• 负责人: MANIKKAM SUTHANTHIRAN
• 金额: $ 83.56万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9128779
• 关键词: Adult; Affect; Albuminuria; Atrophic; Biological; Biological Assay; Biological Markers; Biopsy; Cancer Prognosis; Cardiovascular Diseases; Caring; Cell physiology; Cessation of life; Childhood; Chronic Kidney Failure; Classification; Clinical; Clinical Trials; Cohort Studies; Collaborations; Complementary DNA; Data; Development; Diagnosis; Dialysis procedure; Disease; Disease Outcome; Disease Progression; Early Diagnosis; End stage renal failure; Etiology; Evaluation; Extracellular Matrix; Functional disorder; Future; Gene Expression Profile; Gene Expression Regulation; Genetic Transcription; Glomerular Filtration Rate; Health; Histologic; Human; Individual; Infection; Injury; Intervention; Kidney; Kidney Diseases; Kidney Failure; Kidney Transplantation; Knowledge; Laboratories; Life Expectancy; Link; Liquid substance; Malignant Neoplasms; Massive Parallel Sequencing; Measures; Methods; MicroRNAs; Morbidity - disease rate; Outcome; Pathway interactions; Patients; Pattern; Phase; Polymerase Chain Reaction; Process; Prognostic Marker; Proteinuria; Protocols documentation; Public Health; Qualifying; RNA; Renal function; Renal glomerular disease; Research; Research Personnel; Reverse Transcription; Risk; Risk Management; Risk Marker; Sampling; Scientist; Serum; Severity of illness; Small RNA; Specimen; Staging; Structure; Subgroup; Techniques; Technology; Testing; Tissues; Transcript; Transplant Recipients; Transplantation; Tubular formation; Untranslated RNA; Urine; Validation; adverse outcome; base; biomarker discovery; candidate marker; clinical biomarkers; cohort; cost; cost effective; deep sequencing; experience; extracellular; glomerulosclerosis; high throughput analysis; improved; innovation; insight; interstitial; kidney allograft; microvesicles; mortality; novel; nuclease; patient subsets; post gamma-globulins; precision medicine; predictive marker; premature; prevent; social; specific biomarkers; tool development; transcriptome; urinary; verification and validation

DESCRIPTION (provided by applicant): Chronic kidney disease (CKD) is responsible for premature death from cardiovascular disease, infections and cancer, considerable human suffering and social costs. A large proportion of CKD patients develop end stage renal disease and require dialysis or kidney transplantation. Identifying patients at risk for CKD progression may facilitate precision medicine and prevent attendant complications. Development of mechanistic biomarkers predictive of CKD progression may help develop new treatment(s). Non-coding RNA (ncRNA), specifically microRNA (miRNA) is increasingly recognized as biomolecules that affect multiple cellular processes via regulation of gene expression. It is known that tissue RNA expression conveys information on disease status. Recently, it has been recognized that extracellular RNA (exRNA) is protected from nucleases by their encompassing microvesicles, and therefore can be measured in biological fluids including urine, and convey pathophysiological knowledge. Our labs have extensive experience in high throughput analysis of RNA expression. We have developed a cDNA deep sequencing method to profile miRNA in multiple samples, in a cost-effective, transcriptome-wide and bias-reduced manner. By studying thousands of diverse human samples we have discovered and curated miRNA and other small RNA. Currently, we are developing a parallel method to screen for all ncRNA. We have also adapted quantitative reverse-transcription polymerase chain reaction (qRT-PCR) technology to allow non- invasive verification and validation in larger study cohorts of patients with native and transplant kidney disease. We propose to apply our methods to comprehensively profile miRNA and other ncRNA in urine specimens collected from CKD patients. We aim to identify exRNA in the urine that can predict progression of CKD and develop prognostic biomarkers. We propose a 2-stage protocol; a discovery stage applying deep sequencing for transcriptome-wide profiling and identification of candidate markers, and a validation phase applying RT- qPCR to characterize additional samples and qualify the discovered findings. To allow rapid implementation of our research plan we established collaborations with investigators conducting longitudinal cohort studies of CKD. Urine specimens, linked with rich demographic, clinical and kidney outcome data are available from studies of both adult and pediatric cohorts; both glomerular and non-glomerular diseases; in both native kidneys and kidney transplant recipients. This permits discovery of biomarkers capable of predicting progression across the spectrum of kidney diseases as mechanisms of CKD progression are to a largely independent of CKD etiology. Nonetheless, our studies are powered for subgroup-specific biomarker discovery. Our preliminary findings confirm feasibility to profile exRNA in biofluids, and to detect associations with clinical relevant outcomes. We predict that our findings will inform basic scientist on RNA dysregulation in kidney disease progression and provide clinical scientist biomarker candidates for clinical trials.

描述（由申请人提供）： 慢性肾脏疾病（CKD）导致心血管疾病，感染和癌症，大量人类痛苦和社会成本过早死亡。很大一部分CKD患者患有终阶段肾脏疾病，需要透析或肾脏移植。识别有CKD进展风险的患者可能有助于精确药物并防止随之而来的并发症。机械生物标志物预测CKD进展可能有助于开发新的治疗方法。非编码RNA（NCRNA），特异性microRNA（miRNA）越来越被认为是通过调节基因表达来影响多个细胞过程的生物分子。众所周知，组织RNA表达传达了有关疾病状况的信息。最近，人们已经认识到，细胞外RNA（EXRNA）通过包含的微囊泡保护了核酸酶，因此可以在包括尿液在内的生物流体中进行测量，并传达病理生理学知识。我们的实验室在RNA表达的高通量分析方面具有丰富的经验。我们已经开发了一种cDNA深度测序方法，以成本效益，整个转录组和偏置方式的方式在多个样品中介绍miRNA。通过研究数千种不同的人类样品，我们发现并策划了miRNA和其他小RNA。当前，我们正在开发一种平行方法来筛选所有NCRNA。我们还改编了定量反向转录聚合酶链反应（QRT-PCR）技术，以允许在较大的患有天然和移植肾脏疾病的患者的较大研究队列中进行非侵入性验证和验证。我们建议将我们的方法应用于从CKD患者收集的尿液标本中全面介绍miRNA和其他NCRNA。我们旨在鉴定尿液中可以预测CKD进展并发展预后生物标志物的ExRNA。我们提出了一个2阶段的协议；一个发现深度测序的发现阶段，用于整个转录组分析和识别候选标记，以及应用RT-qPCR的验证阶段来表征其他样本并符合发现的发现。为了允许快速实施我们的研究计划，我们与进行CKD纵向队列研究的研究人员建立了合作。尿液标本与人群，临床和肾脏结局数据有关，可从成人和小儿同伙的研究中获得。肾小球和非斜体疾病；在本地肾脏和肾脏移植受者中。这允许发现能够预测肾脏疾病频谱进展的生物标志物，因为CKD进展的机制主要与CKD病因无关。尽管如此，我们的研究仍用于亚组特异性生物标志物发现。我们的初步发现证实了生物流体中谱系exrna的可行性，并检测与临床相关结果的关联。我们预测，我们的发现将为基础科学家提供有关肾脏疾病进展中RNA失调的基础科学家，并为临床科学家生物标志物候选者提供临床试验。