Single-nucleus sequencing and in situ mapping of mRNA molecules in human kidney

人肾 mRNA 分子的单核测序和原位作图

基本信息

批准号：
9565411
负责人：
Sanjay Jain
金额：
$ 45.41万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-15 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9565411
关键词：
Acute Adult Age Anatomy Archives Atlases Attention Biological Markers Biology Biopsy Catalogs Cell Nucleus Cells Chronic Kidney Failure Classification Clinical assessments Collection Communities Complex Consent Cytolysis Data Data Analyses Data Collection Data Set Development Diagnosis Disease Ensure Environment Etiology Evaluation Freezing Genes Genetic Glean Health Heterogeneity Histologic Human In Situ Injury Kidney Kidney Diseases Knowledge Laboratories Maps Messenger RNA Methods Molecular Molecular Profiling Nephrectomy Nephrons Nuclear Organ Pathologic Pathology Patients Phase Physiology Publications Quality Control RNA Race Renal function Reporting Reproducibility Research Infrastructure Resolution Sampling Specimen Specimen Handling Stress Structure Technology Testing Tissue Preservation Tissues Transcript Translational Research Twin Multiple Birth Universities Urine Variant Washington base cell type experimental study high throughput technology imaging modality improved in vivo insight kidney cell molecular phenotype novel outcome forecast preservation repository sex single cell technology submicron targeted treatment transcriptome sequencing

项目摘要

Abstract The human kidney is a complex organ reported to contain over 25 cell types spatially organized into a million nephrons connected to a urine-transporting network. However, the actual cellular heterogeneity is likely to be much greater. Together, kidney structures function in heterogeneous environments created by the distinct organization (cortical- medullary) and distribution of distinct cell types. Genetic differences, injury and stress can result in kidney disease. A large gap exists in our understanding of the molecular blueprint that drives homeostatic interactions or pathology in kidney. A complete catalog of molecular signatures of all kidney cell types in 3D space, with single cell resolution, is crucial for an improved understanding of normal kidney functions, and acute and chronic kidney disease (AKD and CKD). In this project we will fill this existing gap with three specific aims. In the UG3 phase, with Aim 1, we will collect, preserve and prepare human kidney specimens from consented patients (existing and new) and optimize a single-nucleus RNA sequencing method to human kidneys. In Aim 2, we will optimize an multiplexed in situ RNA imaging method for spatially mapping the transcripts of 100+ genes within human kidney. In the UH3 phase (Aim 3), we will generate a reference cell atlas of human kidney with cell types, sub-types and their relative composition, a true 3D sub-micron resolution spatial map, and apply this knowledge to uncover molecular differences at a cellular level in CKD

抽象的人类肾脏是一种复杂的器官，据报道在空间上包含25种细胞类型组织成与尿液传输网络相连的一百万个肾词。然而，实际的细胞异质性可能会更大。在一起，肾脏结构在不同组织创建的异质环境中的功能（皮质 - 髓质）和不同细胞类型的分布。遗传差异，伤害和压力可能导致肾脏疾病。我们对分子的理解存在很大的差距驱动肾脏稳态相互作用或病理学的蓝图。完整 3D空间中所有肾细胞类型的分子特征的目录，单细胞解决方案对于改善对正常肾脏功能的理解至关重要，并且急性和慢性肾脏疾病（AKD和CKD）。在这个项目中，我们将填写这个现有的差距具有三个特定目标。在UG3阶段，使用AIM 1，我们将收集，保存和从同意的患者（现有和新）和优化人肾脏的单核RNA测序方法。在AIM 2中，我们将优化一种多路复用的原位RNA成像方法，用于空间映射人类肾脏中100多个基因的转录本。在UH3阶段（AIM 3），我们将生成具有细胞类型，亚型及其的人肾脏的参考细胞图集相对组成，真实的3D子微调分辨率空间图，并应用此知识以发现CKD中细胞水平的分子差异