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.

与非裔美国人相比，非裔美国人在肾移植后发生移植肾丢失的风险更高 其他种族。我们的肾移植功能恶化基因组研究(DeKAF：U19 AI070119)显示 慢性移植物功能障碍(CGD)患者首次活检术后移植物丢失情况在两组间存在显著差异 和非AAA。DeKAF的研究从2005年到2011年招募了近3000名移植接受者，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 用于长期临床数据的系统(USRDS)。我们通过自我报告定义了AA，并得到了GWAS校长的验证 组件。CGD的定义是肌酐与3个月基线相比增加了25%， 与同种异体移植物丢失有关。虽然91%的CGD活检有炎症，但并不是所有的都进展到同种异体移植 损失。再生障碍性贫血和非再生障碍性贫血CGD活检之间的分子和细胞差异尚不清楚； 可以利用DeKAF队列中的数据来确定这些差异。解决肾移植丢失的问题 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系。由于巨噬细胞和自然杀手的角色已经确立 (NK)细胞在同种异体移植排斥反应中，中心假说是这些细胞类型在AA中具有更高的丰度。 CGD活检和这些细胞类型将增加移植肾丢失的风险。我们会 测定CGD移植肾活检中巨噬细胞(AIM 1)和NK细胞(AIM 2)丰度的差异 AAs和On-AAs之间的关系以及与同种异体移植物丢失的关系。我们将使用数字空间剖面图(DSP) 确定再生障碍性贫血和非再生障碍性贫血CGD活检的区别。数字信号处理器结合了组织的荧光成像 结构和整个转录组图谱。DSPs具有创新性，因为它区分了RNA和蛋白质 在不同的组织中表达，如肾小管、间质或肾小球。其原因是 DSP法：组织学显示移植肾间质区纤维化增多且增多 来自DeKAF的AA CGD活检中的肾小管萎缩，但不是什么细胞类型与肾脏损害有关。 我们将评估4组中每一组的CGD活检：1)有同种异体移植物丢失的AAS 2)无同种异体移植物丢失的AAS 3) 移植物丢失的非腹主动脉和不伴移植物丢失的非腹主动脉。我们预计会发现更丰富的 巨噬细胞和NK细胞与AAS和同种异体移植物丢失相关。这项提议将开发创新的 DSP技术评估CGD活检导致分子、细胞和空间的定义 腹主动脉和非腹主动脉肾移植的差异及其与移植物丢失的关系研究。这项研究将引领 创建各种细胞类型的空间图谱和再生障碍性贫血和非再生障碍性贫血活检组织中的转录本 同种异体移植丢失。这项研究应该开发丰富的数据，作为机理和 干预性R01建议遵循。我们设想DSP补充病理以指导个性化 治疗CGD，并有助于缩小AA移植结果差异的差距。