Integrated spatial interrogation of cellular and molecular signatures of human kidney disease

(请保存在Word中，不要保存为PDF) 在此输入文本，它是您的应用程序的新摘要信息。此部分不得超过30行文本。 摘要 在理解决定人类肾脏疾病进展的机制方面存在着根本性的差距。长期目标是确定调控急性和慢性肾脏疾病(AKI和CKD)进展的关键细胞和分子途径，识别评估疾病进展的新标记物，并针对这些途径开发特定的治疗干预措施。肾脏损伤的模式通常是不同的，不同的细胞类型和细胞状态不同地分布在活检标本的焦点区域。在这些损伤区域观察到免疫细胞亚群，表明浸润性免疫细胞与常驻的肾上皮细胞、内皮细胞和间质细胞之间存在强烈的相互作用。此外，在标准的组织病理学评估中，这些区域性分子疾病的表型往往没有得到充分的认识。因此，迫切需要扩大我们的努力，全面定位肾脏的细胞类型和细胞状态，亚表型疾病，揭示细胞与细胞之间的相互作用，并了解肾脏形态变化背后的细胞和分子表型。这一建议的首要假设是，只有在个体肾脏的空间背景下解释细胞和分子表型的多样性时，才能理解肾脏疾病的机制。中心假说将使用空间分辨成像、转录和蛋白质组学技术的集成管道进行验证，这些技术将在细胞、区域和器官水平上捕捉肾脏疾病的特征。这些技术包括大规模3D显微镜和组织细胞术(3DTC)、空间转录(ST)和CODEX多路免疫荧光检测。总之，这一分析管道将有助于绘制损伤分布图，并通过以下目标与其他KPMP技术有效地联系起来：具体目标1：部署一套KPMP批准的空间技术作为同步分析管道，以确定每个人类肾脏活检的细胞成分和分子结构。具体目标2：整合来自每个活检标本的成像、转录和蛋白质组特征，以生成细胞和分子表型的多组分空间图。具体目标3：整合我们跨KPMP的分析管道，将我们空间锚定的细胞和分子表型与其他KPMP技术、病理学和临床特征联系起来。这项拟议的研究意义重大，因为它是一系列研究的下一步，预计将确定疾病进展的迫切需要的生物标记物，优化临床前研究，并在AKI和CKD这一巨大的临床问题上开发特定和有针对性的治疗干预措施。

(PLEASE KEEP IN WORD, DO NOT PDF) Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Abstract There is a fundamental gap in understanding the mechanisms that determine progression in human kidney disease. The long-term goal is to characterize key cellular and molecular pathways regulating progression of acute and chronic kidney diseases (AKI and CKD), to identify novel markers that assess disease progression, and to develop specific therapeutic interventions targeting these pathways. There is often heterogeneity in the pattern of kidney injury whereby various cell types and cell states are differentially distributed across focal regions of a biopsy specimen. Immune cell sub-populations are observed in these injury regions, suggesting robust cross-talk between infiltrating immune cells and the resident renal epithelial, endothelial, and stromal cells. Further, these regional molecular disease phenotypes often are not fully appreciated on standard histopathologic assessment. Therefore, there is a critical need to expand our efforts to comprehensively localize cell types and cell states in the kidney, sub-phenotype disease, uncover cell-cell interactions, and understand the cellular and molecular phenotypes underlying morphologic changes in the kidney. The overarching hypothesis of this proposal is that mechanisms of kidney disease can only be understood when the diversity of cellular and molecular phenotypes are interpreted in the spatial context of the individuals’ kidney. The central hypothesis will be tested using an integrated pipeline of spatially resolved imaging, transcriptomic, and proteomic technologies that will capture the features of kidney disease at the cell, regional, and organ levels. These technologies include large scale 3D microscopy and tissue cytometry (3DTC), spatial transcriptomics (ST), and CO-Detection by indEXing (CODEX) multiplexed immunofluorescence. Together, this analytical pipeline will facilitate mapping of injury distribution and effectively link with other KPMP technologies through the following aims: Specific Aim 1: Deploy a suite of KPMP-approved spatial technologies as a synchronized analytical pipeline to define the cellular components and molecular structure of each human kidney biopsy. Specific Aim 2: Integrate imaging, transcriptomic, and proteomic signatures from each biopsy specimen to generate a multi-component spatial map of cellular and molecular phenotypes. Specific Aim 3: Integrate our analytical pipeline across the KPMP, linking our spatially-anchored cellular and molecular phenotypes to other KPMP technologies, pathology, and clinical features. The proposed research is significant, because it is the next step in a continuum of research that is expected to identify critically needed biomarkers of disease progression, optimize preclinical studies, and develop specific and targeted therapeutic interventions in the vast clinical problem of AKI and CKD.