Spatial Elucidation of Human Acute Kidney Injury and Chronic Kidney Disease using Imaging Mass Cytometry

使用成像质谱流式细胞术空间阐明人类急性肾损伤和慢性肾病

基本信息

批准号：
10701865
负责人：
LLOYD G CANTLEY
金额：
$ 36万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10701865
关键词：
3-Dimensional Ablation Acute Renal Failure with Renal Papillary Necrosis Affect Altruism Animal Model Antibodies Attention Automobile Driving Award Binding Biopsy Biopsy Specimen Cell Communication Cell Count Cells Cessation of life Chronic Kidney Failure Clinical Compensation Complex Coupled Critical Illness Cytometry Data Data Analyses Ensure Etiology Formalin Freezing Generations Heavy Metals Homing Human Image Individual Infiltration Injury Injury to Kidney Intervention Kidney Kidney Diseases Knowledge Lasers Location Machine Learning Maps Mass Spectrum Analysis Molecular Monoclonal Antibodies Nuclear Nuclear RNA Paraffin Embedding Pathogenesis Pathogenicity Pathway interactions Patients Performance Persons Pharmacological Treatment Physiologic pulse Population Process Proteomics Protocols documentation Quality Control Rare Earth Metals Reproducibility Resolution Risk Running Sampling Signal Transduction Spottings Standardization Syndrome System T-Lymphocyte Subsets Techniques Technology Time Tissues Tubular formation Work analysis pipeline antibody conjugate cell injury cell type computerized data processing delayed graft function dimensional analysis eosinophil glomerular filtration high dimensionality improved improved outcome injury and repair kidney biopsy kidney cell kidney repair mast cell member metabolomics mortality neutrophil novel precision medicine preimplantation preservation prevent prototype quality assurance reconstruction recruit response single cell sequencing therapeutic target transcriptome sequencing tumor vascular injury

项目摘要

Abstract Acute kidney injury (AKI) is a syndromic term encompassing a wide range of insults and pathogenic responses that lead to a rapid reduction in glomerular filtration. Although we have made significant progress in our understanding of kidney injury in animal models, far less attention has been focused on the pathogenesis and treatment of the diverse types of human AKI. The largest barrier in achieving this knowledge is the limited number of kidney biopsies performed for AKI and the small amount of tissue obtained from renal biopsy. The Kidney Precision Medicine Project is tackling this complex issue by recruiting altruistic patients with AKI who are willing to have kidney biopsies in order to advance our knowledge of human kidney disease. These biopsies are being interrogated using multiple complementary technologies. We propose to use Imaging Mass Cytometry to help provide a highly detailed, quantitative cellular map of nearly all cells present in sections from those kidney biopsies, including their differentiation state and activation of injury and repair pathways. Combined with the cell sequencing, metabolomic, and proteomic data generated under KPMP guidance, this will provide a substantial increase in our understanding of human AKI and CKD. Imaging mass cytometry (IMC) uses a high-resolution laser combined with a mass cytometer to detect the presence, location and amount of up to 42 different heavy metal conjugated antibodies hybridized to a tissue section. We have successfully used IMC with a panel of 22 heavy metal conjugated validated antibodies to identify resident kidney cell populations, infiltrating cell populations, and cell activation and injury states using archival FFPE human kidney tissue, and developed a machine learning technique termed Kidney MAPPS to rapidly and accurately identify, quantify and localize ~92% of all cells in those biopsies. We now propose to increase that validated antibody panel to >30 antibodies that will allow identification of >95% of cells and improve cell injury and activation state assessment, and to optimize the IMC and Kidney-MAPPS analysis pipeline to perform 2D and 3D quantitative assessment of cell location, cell-cell interactions and cellular responses in human AKI and CKD biopsy tissues (SA1). We will standardize a defined work-flow protocol coupled with rigorous quality control assessment steps at key points (SA2), and then apply this IMC work-flow to kidney samples provided by KPMP and integrate with the KPMP Central Hub and consortium members to develop accurate protocols for mapping the scRNAseq/snRNAseq data, proteomics data and metabolomics data onto the appropriate cells and locations using Kidney-MAPPS (SA3).

抽象的急性肾损伤 (AKI) 是一个综合征术语，涵盖多种损伤和致病因素导致肾小球滤过迅速减少的反应。尽管我们已经取得了重大成果我们对动物模型肾损伤的理解取得了进展，但很少有人关注不同类型人类 AKI 的发病机制和治疗。实现这一目标的最大障碍知识是针对 AKI 进行的肾活检数量有限且组织数量较少从肾活检获得。肾脏精准医学项目正在解决这个复杂的问题招募愿意进行肾活检的利他性 AKI 患者，以推进我们的研究人类肾脏疾病的知识。这些活组织检查正在使用多种互补的方法进行询问技术。我们建议使用成像质谱流式细胞仪来帮助提供高度详细的定量信息肾活检切片中几乎所有细胞的细胞图谱，包括它们的分化损伤和修复途径的状态和激活。结合细胞测序、代谢组学和在 KPMP 指导下生成的蛋白质组数据，这将大幅提高我们的了解人类 AKI 和 CKD。成像质谱流式细胞术 (IMC) 使用高分辨率激光与质谱流式细胞仪相结合来检测多达 42 种不同的重金属偶联抗体的存在、位置和数量，这些抗体与组织切片。我们已成功将 IMC 与经过验证的 22 种重金属缀合组一起使用用于识别常驻肾细胞群、浸润细胞群和细胞活化的抗体使用档案 FFPE 人类肾脏组织来研究损伤状态，并开发了一种机器学习技术称为“肾脏 MAPPS”，可快速准确地识别、量化和定位肾脏中约 92% 的细胞活检。我们现在建议将经过验证的抗体组增加到超过 30 种抗体，这将允许识别 >95% 的细胞并改善细胞损伤和激活状态评估，并优化 IMC 和 Kidney-MAPPS 分析流程可对细胞位置进行 2D 和 3D 定量评估，人类 AKI 和 CKD 活检组织 (SA1) 中的细胞间相互作用和细胞反应。我们将将定义的工作流程协议标准化，并在关键环节加上严格的质量控制评估步骤点（SA2），然后将此 IMC 工作流程应用于 KPMP 提供的肾脏样本并与 KPMP 中央中心和财团成员制定准确的协议来绘制地图 scRNAseq/snRNAseq 数据、蛋白质组学数据和代谢组学数据转移到适当的细胞上并使用 Kidney-MAPPS (SA3) 的位置。