Molecular and Cellular Analysis of Allograft Loss in Kidney Transplant Biopsies

肾移植活检中同种异体移植物丢失的分子和细胞分析

• 批准号: 10511352
• 负责人: Casey R Dorr
• 金额: $ 24.75万
• 依托单位: HENNEPIN HEALTHCARE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10511352
• 关键词: Address; African American population; Alabama; Allografting; Antibodies; Area; Atlases; Atrophic; Biological; Biological Markers; Biopsy; Cell Survival; Chronic; Clinical; Clinical Data; Confounding Factors (Epidemiology); County; Creatinine; Data; Deterioration; Diagnosis; Dose; Enrollment; Fibrosis; Foundations; Functional disorder; Genetic Transcription; Genomics; Histologic; Histology; Immunosuppression; Inflammation; Information Systems; Intervention; Kidney; Kidney Transplantation; Lead; Link; Location; Medical center; Minnesota; Mission; Molecular; Morphology; NCAM1 gene; Natural Killer Cells; Outcome; Participant; Pathologist; Pathology; Patient Self-Report; Public Health; RNA; Race; Research; Risk; Risk Assessment; Role; Socioeconomic Factors; Structure; Technology; Tissue imaging; Tissues; Training; Transcript; Transplant Recipients; Transplantation; Tubular formation; United States; United States National Institutes of Health; Universities; Update; Variant; allograft rejection; cell type; cellular imaging; cohort; digital; fluorescence imaging; follow-up; genome wide association study; high risk; improved; innovation; interstitial; kidney allograft; kidney dysfunction; macrophage; melanoma; multidisciplinary; personalized medicine; programmed cell death ligand 1; protein expression; renal damage; single-cell RNA sequencing; transcriptome; transcriptome sequencing; transplant centers

African Americans (AAs) have higher risk for kidney allograft loss after kidney transplantation compared with other races. Our Deterioration of Kidney Allograft Function Genomics study (DeKAF: U19 AI070119) showed a significant disparity in allograft loss after the first biopsy for chronic allograft dysfunction (CGD) between AAs and non-AAs. The DeKAF study enrolled nearly 3,000 transplant recipients, from 2005 to 2011, and conducted genome wide association studies (GWAS). DeKAF participants are linked to the United States Renal Data System (USRDS) for long-term clinical data. We defined AA by self-report and verified by GWAS principal components. CGD was defined as >25% increase in creatinine relative to a 3-month baseline and is associated with allograft loss. Although 91% of CGD biopsies have inflammation, not all progress to allograft loss. The molecular and cellular differences between AA and non-AA CGD biopsies are unknown; biopsies from the DeKAF cohort can be leveraged to determine these differences. To address kidney allograft loss disparities after CGD, we aim to determine the molecular and cellular differences between AA and non-AA CGD biopsies and associations with allograft loss. Due to established roles for macrophages and natural killer (NK) cells in allograft rejection, the central hypothesis is that these cell types have higher abundance in AA CGD biopsies and these cell types will be associated with increased risk for kidney allograft loss. We will determine differences in Macrophage (Aim 1) and NK Cell (Aim 2) abundance in CGD kidney allograft biopsies between AAs and on-AAs and association with allograft loss. We will use Digital Spatial Profiling (DSP) to determine differences between AA and non-AA CGD biopsies. DSP combines fluorescent imaging of tissue structures and whole transcriptome profiling. DSP is innovative because it differentiates RNA and protein expression in separate tissue compartments such as tubules, interstitium or glomerulus. The reason for the DSP approach: Histology showed more fibrosis in the interstitial areas of the kidney allografts and increased tubular atrophy in AA CGD biopsies from DeKAF, but not what cell types are associated with kidney damage. We will evaluate CGD biopsies in each of 4 groups: 1) AAs with allograft loss 2) AAs without allograft loss 3) non-AAs with allograft loss and 4) non-AAs without allograft loss. We expect to find higher abundance of macrophages and NK cells associated with AAs and allograft loss. This proposal will develop the innovative DSP technology to assess CGD biopsies leading to a definition of the molecular, cellular and spatial differences in AAs and non-AAs kidney allografts and study associations with allograft loss. This study will lead to creation of a spatial atlas of various cell types and transcripts in AA and non-AA biopsies that associate with allograft loss. This study should develop a wealth of data, as the foundation for a mechanistic and interventional R01 proposal to follow. We envision DSP supplementing pathology to guide personalized therapy for CGD and help close the gap in AA transplant outcome disparities.

非洲裔美国人（AAS）与肾移植后肾脏同种异体丧失的风险更高 其他种族。我们对肾脏同种异体移植功能基因组学研究的恶化（DEKAF：U19 AI070119）显示出一个 AAS慢性同种异体功能障碍（CGD）的第一次活检后同种异体移植丧失的显着差异 和非AAS。 DEKAF研究从2005年到2011年招募了近3,000名移植受者，并进行了 基因组广泛的关联研究（GWAS）。 DEKAF参与者链接到美国肾脏数据 长期临床数据的系统（USRD）。我们通过自我报告定义了AA，并由GWAS校长验证 成分。 CGD定义为肌酐相对于3个月的基线增长> 25％，IS 与同种异体丧失有关。尽管有91％的CGD活检患有炎症，但并非所有人都会进展到同种异体移植 损失。 AA和非AA CGD活检之间的分子和细胞差异尚不清楚。活检 可以从DEKAF队列中利用DEKAF队列来确定这些差异。解决肾脏同种异体损失 CGD之后的差异，我们旨在确定AA和非AA之间的分子和细胞差异 CGD活检和与同种异体丧失的关联。由于巨噬细胞和自然杀手的既定角色 （NK）同种异体移植排斥的细胞，中心假设是这些细胞类型的丰度更高 CGD活检和这些细胞类型将与肾脏同种异体移植丧失的风险增加有关。我们将 确定巨噬细胞的差异（AIM 1）和NK细胞（AIM 2）CGD肾脏同种异体移植活检中的丰度 在AAS和AAS之间以及与同种异体移植损失的关联之间。我们将使用数字空间分析（DSP） 确定AA和非AA CGD活检之间的差异。 DSP结合组织的荧光成像 结构和整个转录组分析。 DSP具有创新性，因为它会区分RNA和蛋白质 在单独的组织室中的表达，例如小管，间质或肾小球。原因 DSP方法：组织学在肾脏同种异体移植物的间隙区域显示更多的纤维化并增加 DEKAF的AA CGD活检中的管状萎缩，但与肾脏损伤无关。 我们将评估四个组中每一部的CGD活检：1）同种异体损失的AA 2）AAS没有同种异体移植损失3） 具有同种异体损失的非AAS和4）非同种异体损失的非AAS。我们希望发现更高的丰度 巨噬细胞和与AAS和同种异体移植损失相关的NK细胞。该建议将发展创新 DSP技术评估CGD活检，导致分子，细胞和空间的定义 AAS和非AAS肾脏同种异体移植物的差异以及同种异体丧失的研究关联。这项研究将领导 在AA和非AA活检中创建各种细胞类型和转录本的空间图集 同种异体移植损失。这项研究应开发大量数据，作为机械和 介入的R01提案要遵循。我们设想DSP补充病理来指导个性化 CGD治疗，并有助于缩小AA移植结果差异的差距。