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: 10628042
• 负责人: Casey R Dorr
• 金额: $ 15.87万
• 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 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10628042
• 关键词: Address; African American population; Alabama; Allografting; Antibodies; Area; Atlases; Atrophic; Biological; Biological Markers; Biopsy; Cd68; Cell Survival; Chronic; Clinical; Clinical Data; Cohort Studies; Confounding Factors (Epidemiology); County; Creatinine; Data; Deterioration; Diagnosis; Disparity; Dose; Enrollment; Fibrosis; Foundations; Functional disorder; Genetic Transcription; Genomics; Histologic; Histology; Immunosuppression; Inflammation; Information Systems; Intervention; Kidney; Kidney Transplantation; Link; Location; Macrophage; Medical center; Micro Array Data; 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; digital; fluorescence imaging; follow-up; genome wide association study; high risk; improved; innovation; interstitial; kidney allograft; kidney dysfunction; melanoma; multidisciplinary; outcome disparities; personalized medicine; programmed cell death ligand 1; protein expression; renal damage; single-cell RNA sequencing; transcriptome; transcriptome sequencing; transcriptomic profiling; 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.

与非洲裔美国人 (AA) 相比，肾移植后同种异体移植肾丢失的风险更高 其他种族。我们的肾同种异体移植功能恶化基因组学研究 (DeKAF: U19 AI070119) 显示 AA 之间因慢性同种异体移植功能障碍 (CGD) 首次活检后同种异体移植物损失存在显着差异 和非AA。 DeKAF 研究从 2005 年到 2011 年招募了近 3,000 名移植受者，并进行了 全基因组关联研究（GWAS）。 DeKAF 参与者链接到美国肾脏数据 用于长期临床数据的系统（USRDS）。我们通过自我报告定义 AA，并由 GWAS 负责人验证 成分。 CGD 定义为肌酐相对 3 个月基线增加 >25%， 与同种异体移植物丢失有关。尽管 91% 的 CGD 活检存在炎症，但并非所有进展都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 可以利用 DeKAF 队列中的数据来确定这些差异。解决同种异体移植肾丢失问题 CGD 后的差异，我们的目标是确定 AA 和非 AA 之间的分子和细胞差异 CGD 活检及其与同种异体移植物丢失的关联。由于巨噬细胞和自然杀伤细胞的既定作用 同种异体移植排斥中的 (NK) 细胞，中心假设是这些细胞类型的 AA 丰度较高 CGD 活检和这些细胞类型将与肾同种异体移植物丢失的风险增加相关。我们将 确定 CGD 肾同种异体移植物活检中巨噬细胞（目标 1）和 NK 细胞（目标 2）丰度的差异 AA 和 on-AA 之间的关系以及与同种异体移植物损失的关系。我们将使用数字空间剖析 (DSP) 确定 AA 和非 AA CGD 活检之间的差异。 DSP 结合组织荧光成像 结构和全转录组分析。 DSP 具有创新性，因为它可以区分 RNA 和蛋白质 在单独的组织区室（例如肾小管、间质或肾小球）中表达。原因是 DSP 方法：组织学显示同种异体肾移植物的间质区域有更多纤维化，并且增加 DeKAF 的 AA CGD 活检中显示肾小管萎缩，但不知道哪些细胞类型与肾脏损伤相关。 我们将评估 4 组中每组的 CGD 活检：1) 具有同种异体移植物丢失的 AA 2) 不具有同种异体移植物丢失的 AA 3) 具有同种异体移植物损失的非AAs和4)无同种异体移植物损失的非AAs。我们期望找到更高丰度的 巨噬细胞和 NK 细胞与 AA 和同种异体移植物损失相关。该提案将开发创新 用于评估 CGD 活检的 DSP 技术，从而确定分子、细胞和空间 AA 和非 AA 肾同种异体移植物的差异以及与同种异体移植物丢失的研究关联。这项研究将引导 创建与 AA 和非 AA 活检中的各种细胞类型和转录本相关的空间图谱 同种异体移植物损失。这项研究应该开发大量数据，作为机械和 遵循干预性 R01 提案。我们设想 DSP 补充病理学以指导个性化治疗 治疗 CGD 并有助于缩小 AA 移植结果差异的差距。