Non-Coding RNA and CKD Progression

项目摘要 肾纤维化是大多数肾损伤下游的最终共同途径，其导致进行性肾损伤。 慢性肾病（CKD）。非编码RNA通过直接抑制和/或调节肾纤维化 基质基因的表达和通过TGF-β信号传导。我们的核心工作假设是， 循环和尿microRNA和长链非编码RNA（EncRNA）是潜在肾纤维化的指标 因此是CKD进展的早期生物标志物。我们将使用慢性肾功能不全队列研究 生物样本和相关数据。该研究将分三个阶段进行：发现、复制 和实验验证阶段。下一代测序将用于分析血液中的ncRNA 以及来自表现出慢性肾病缓慢（n=191）和快速（n=192）进展的参与者的尿样。顶部CKD 进展相关的ncRNA发现将在3,088名CRIC研究参与者中使用定量RT- PCR法关注的主要肾脏表型是估计的肾小球滤过率的变化斜率 （eGFR），以及至终末期肾病（ESRD）的时间，或至ESRD或降低50%的时间的复合终点 eGFR。我们将采用创新的体外实验和体内功能获得和丧失实验， 小鼠模型，以识别靶点并验证人类EncRNA发现的功能意义 问题研究最后，我们将验证在显微解剖的CKD中发现的最高EncRNA的表达水平。 肾脏组织确定CKD进展的特异性ncRNA途径将增强诊断， 使风险分层，并导致假设驱动的有针对性的干预措施。

Project Summary Kidney fibrosis is the final common pathway downstream of most renal injuries that contributes to progressive chronic kidney disease (CKD). Non-coding RNAs regulate kidney fibrosis through direct repression and/or expression of matrix genes and through TGF-β signaling. Our central working hypothesis is that specific circulating and urinary microRNA and long non-coding RNA (EncRNA) are indicators of underlying kidney fibrosis and hence are early biomarkers for CKD progression. We will use the Chronic Renal Insufficiency Cohort Study (CRIC) biosamples and associated data. The study will be conducted in three phases; discovery, replication and an experimental validation phases. Next-generation sequencing will be used to profile the ncRNAs in blood and urine samples from participants exhibiting slow (n=191) and fast (n=192) progression of CKD. Top CKD progression-related ncRNA discoveries will be replicated in 3,088 CRIC study participants using quantitative RT- PCR. The primary renal phenotypes of interest are the slope of change in estimated glomerular filtration rate (eGFR), and either time to end-stage kidney disease (ESRD), or a composite of time to ESRD or a 50% reduction in eGFR. We will employ innovative in vitro experiments and in vivo gain and loss of function experiments in mouse models to identify targets and validate the functional significance of the EncRNA discoveries from human studies. Finally, we will verify the expression level of the top EncRNA discoveries in the microdissected CKD kidney tissues. Identification of specific ncRNA pathways for the progression of CKD will enhance diagnosis, enable risk stratification and lead to hypothesis driven targeted interventions.