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

• 批准号: 10515143
• 负责人: LLOYD G CANTLEY
• 金额: $ 36万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 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/10515143
• 关键词: 3-Dimensional; Acute Renal Failure with Renal Papillary Necrosis; Affect; Altruism; Animal Model; Antibodies; Archives; Attention; Automobile Driving; Award; Biopsy; Biopsy Specimen; Cell Communication; Cell Count; Cells; Cessation of life; Chronic Kidney Failure; Clinical; Complex; Coupled; Critical Illness; Cytometry; Data; Data Analyses; Ensure; Etiology; Financial compensation; Formalin; Freezing; Generations; Heavy Metals; Homing; Human; Hybrids; Image; Individual; Injury; Injury to Kidney; Intervention; Kidney; Kidney Diseases; Knowledge; Lasers; Lead; 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; base; cell injury; cell type; computerized data processing; dimensional analysis; eosinophil; glomerular filtration; high dimensionality; hybrid antibody; improved; improved outcome; injury and repair; kidney biopsy; kidney cell; kidney repair; mast cell; member; metabolomics; neutrophil; novel; precision medicine; preimplantation; preservation; prevent; prototype; quality assurance; reconstruction; recruit; response; single cell sequencing; therapeutic target; transcriptome sequencing; tumor; vascular injury

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).

摘要 急性肾损伤（阿基）是一种综合征术语，包括广泛的损伤和致病性损伤。 导致肾小球滤过率迅速降低的反应。尽管我们已经做出了重大的 随着我们对动物模型中肾损伤的了解取得进展，人们对肾损伤的关注却少得多 不同类型人类阿基的发病机制和治疗。实现这一目标的最大障碍是 知识是有限的数量进行肾活检阿基和少量的组织 从肾活检中获得。肾脏精准医学项目正在解决这个复杂的问题， 招募愿意接受肾活检的利他主义阿基患者，以促进我们的研究。 人类肾脏疾病的知识。这些活组织检查正在使用多个互补的 技术.我们建议使用成像质量细胞计数，以帮助提供一个高度详细的，定量的， 肾脏活检切片中几乎所有细胞的细胞图谱，包括其分化 损伤和修复途径的状态和激活。结合细胞测序，代谢组学， 在KPMP指导下生成的蛋白质组数据，这将大大增加我们的 了解人类阿基和CKD。 成像质谱细胞仪（IMC）使用高分辨率激光与质谱细胞仪相结合，以检测 最多42种不同的重金属缀合抗体的存在、位置和量， 组织切片我们已经成功地使用IMC与一组22个重金属结合验证 抗体，以鉴定常驻肾细胞群、浸润细胞群和细胞活化， 损伤状态使用档案FFPE人类肾脏组织，并开发了机器学习技术 称为肾脏MAPPS，可以快速准确地识别，定量和定位这些细胞中约92%的细胞。 活组织检查我们现在建议将经验证的抗体组增加到>30种抗体， 鉴定>95%的细胞并改善细胞损伤和活化状态评估，并优化 IMC和Kidney-MAPPS分析管道可对细胞位置进行2D和3D定量评估， 人阿基和CKD活检组织中的细胞-细胞相互作用和细胞应答（SA 1）。我们将 标准化已定义的工作流程协议，并在关键环节执行严格的质量控制评估步骤 点（SA 2），然后将该IMC工作流程应用于KPMP提供的肾脏样本，并与 KPMP中央枢纽和联盟成员制定准确的协议， 将scRNAseq/snRNAseq数据、蛋白质组学数据和代谢组学数据导入适当的细胞， 使用Kidney-MAPPS（SA 3）定位。