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.

非裔美国人（AA）在肾移植后发生肾移植物丢失的风险高于 其他种族。我们的肾移植功能恶化基因组学研究（DeKAF：U19 AI 070119）显示， AA之间因慢性同种异体移植物功能障碍（CGD）首次活检后同种异体移植物丢失的显著差异 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参与者与美国肾脏数据相关联 长期临床数据系统（USRDS）。我们通过自我报告定义AA，并通过GWAS负责人验证 件. CGD定义为相对于3个月基线肌酐增加>25%， 与同种异体移植物丢失有关。尽管91%的CGD活检有炎症，但并非所有的CGD活检都进展为同种异体移植物。 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。解决肾移植物丢失问题 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活检与同种异体移植物丢失的相关性。由于巨噬细胞和自然杀伤细胞 (NK)细胞在同种异体移植排斥反应中的作用，中心假设是这些细胞类型在AA中具有更高的丰度， CGD活检和这些细胞类型将与肾移植物丢失的风险增加相关。我们将 确定CGD肾移植活检中巨噬细胞（Aim 1）和NK细胞（Aim 2）丰度的差异 AAs和非AAs之间的差异以及与同种异体移植物丢失的相关性。我们将使用数字空间轮廓（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是创新的，因为它区分了RNA和蛋白质 在单独的组织隔室如小管、肾小球或肾小球中表达。的原因 DSP方法：组织学显示肾移植物间质区纤维化更多， 在DeKAF的AA CGD活检中发现肾小管萎缩，但不知道哪些细胞类型与肾损伤相关。 我们将评估4组中的CGD活检：1）有同种异体移植物丢失的AA 2）无同种异体移植物丢失的AA 3） 非AAs伴同种异体移植物丢失和4）非AAs无同种异体移植物丢失。我们希望能找到更高丰度的 与AA和同种异体移植物丢失相关的巨噬细胞和NK细胞。该提案将发展创新的 DSP技术用于评估CGD活检，从而定义分子、细胞和空间 AAs和非AAs肾同种异体移植物的差异以及与同种异体移植物丢失的相关性研究。这项研究将导致 在AA和非AA活检组织中创建各种细胞类型和转录物的空间图谱， 同种异体移植物丢失。这项研究应该开发出丰富的数据，作为一个机械和 后续介入性R 01提案。我们设想DSP补充病理学，以指导个性化 治疗CGD，并帮助缩小AA移植结果差异的差距。