Epigenetic Regulation of Regulatory B Cell Cytokine Expression and Allograft Rejection

调节性 B 细胞细胞因子表达和同种异体移植排斥的表观遗传调控

• 批准号: 10349822
• 负责人: Aravind Cherukuri
• 金额: $ 18.61万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-17 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10349822
• 关键词: ATAC-seq; Acute; Adult; Advisory Committees; Affect; Allografting; Autoimmune; B-Lymphocyte Subsets; B-Lymphocytes; Basic Science; Bioinformatics; Biological Markers; Biology; Biopsy; Cells; Characteristics; Chronic; Clinical; Clinical Sciences; Clinical Trials Design; Cytokine Gene; DNA; Diagnosis; Dialysis procedure; End stage renal failure; Epigenetic Process; Exhibits; Fibrosis; Gene Expression; Genes; Genetic; Genetic Crossing Over; Genetic Polymorphism; Genetic Transcription; Genetic Variation; Genomics; Goals; Graft Survival; Human; Human Genetics; Immune; Immune response; Immunologics; Immunology; Immunosuppression; Individual; Inflammation; Injury; Interleukin-10; International; Kidney Transplantation; Lead; Maps; Mentors; Monitor; Multicenter Studies; Outcome; Patient risk; Patients; Physicians; Predictive Value; Records; Research; Research Personnel; Risk; Risk Assessment; Scientist; Single Nucleotide Polymorphism Map; Statistical Models; System; TNF gene; Techniques; Testing; Time; Training; Transcriptional Regulation; Translational Research; Transplant Recipients; Transplantation; Transplantation Immunology; allograft rejection; antimicrobial; bisulfite sequencing; career; cytokine; demographics; differential expression; epigenetic regulation; high risk; individual patient; insight; kidney allograft; novel; patient stratification; peripheral blood; post-transplant; predictive marker; prevent; prospective; recruit; response; risk stratification; skills; tenure track; therapeutic target; transcriptome; transcriptome sequencing; translational applications; treatment choice; tumor; unsupervised learning

PROJECT SUMMARY/ABSTRACT Despite remarkable short-term outcomes, the rate of late renal allograft loss has not improved. This is increasingly attributed to the cumulative effects of ongoing subtle immunological injury that often remains undetected. Clinical parameters and even surveillance biopsies have limited predictive value, underscoring the need for new predictive biomarkers. Dr. Cherukuri showed that: (i) Human Breg activity is best characterized by the ratio of IL-10/TNFα expression by immature T1 transitional B cells (T1Bs); and (ii) This T1B IL-10/TNFα ratio 3 mos post-transplant, predicts subsequent acute rejection (AR), allograft fibrosis, decreased eGFR and graft survival. Furthermore, assessment of T1B IL-10/TNFα ratio prior to transplantation showed that in ~75% of patients, the cytokine ratio remains stable after transplantation and predicts their risk for AR and subsequent outcomes (pre-determined). In contrast, in ~25% of patients, the cytokine ratio shifts from high to low or from low to high, and their risk for subsequent outcomes tracks with the direction of the shift. The mechanisms that regulate stable vs. switchable T1B cytokine expression are unknown. Understanding such mechanisms will allow us to accurately risk-stratify patients prior to transplantation and elucidate key regulatory mechanisms that contribute to an individual patient’s underlying immunological reactivity. We hypothesize that transcriptional regulation by genetic or epigenetic mechanisms dictate the stability of the T1B cytokine ratio, Breg activity, and determine a patient’s post-transplant clinical course. This hypothesis will be tested by examining the genetic and epigenetic underpinnings of stable vs. switchable T1B cytokine expression, and by determining whether clinical characteristics correlate with these changes. These Aims will allow Dr. Cherukuri to gain expertise in: (i) state of the art advanced genomic and epigenetic analysis approaches (e.g., ATACseq, bisulfite sequencing, RNAseq), (ii) bioinformatics, and (iii) traditional statistical modelling, and unsupervised machine learning approaches; in addition to honing standard lab techniques. These new indispensable approaches for basic and translational research will facilitate his transition to a career performing meaningful independent research. These skills will be developed under the guidance of a team of mentors, advisors, and collaborators at the Univ. of Pittsburgh. Dr. David Rothstein (1° mentor) is an established physician-scientist with expertise in transplant immunology, tolerance, and Bregs, as well as biomarker research. Dr. Harinder Singh (co-mentor), an internationally recognized scientist with expertise in genetic and epigenetic regulation of immune responses, directs the Univ. of Pittsburgh Center for Systems Immunology. Both mentors have excellent training track records. An advisory committee of accomplished investigators with expertise ranging from fundamental transplant immunology to translational and clinical science will monitor Dr. Cherukuri’s progress biannually. This proposal will promote Dr. Cherukuri’s goal of achieving scientific independence as a tenure track physician-scientist with expertise in Bregs, their transcriptional regulation and translational applications as biomarkers, and as therapeutic targets.

项目概要/摘要 尽管短期效果显着，但晚期同种异体移植肾丢失率并未改善。这是 越来越多地归因于持续的微妙免疫损伤的累积效应，这种损伤通常仍然存在 未被发现。临床参数甚至监测活检的预测价值有限，强调了 需要新的预测生物标志物。 Cherukuri 博士表明： (i) 人类 Breg 活动的最佳特征是 未成熟 T1 移行 B 细胞 (T1B) 表达 IL-10/TNFα 的比率； (ii) T1B IL-10/TNFα 比率 移植后 3 个月，预测随后的急性排斥反应 (AR)、同种异体移植物纤维化、eGFR 下降和移植物 生存。此外，移植前对 T1B IL-10/TNFα 比率的评估表明，约 75% 的患者 对于患者，细胞因子比率在移植后保持稳定，并预测他们发生 AR 和随后的风险 结果（预先确定）。相比之下，约 25% 的患者细胞因子比率从高变为低或从低变化 到高，并且他们后续结果的风险随着转变的方向而变化。的机制 调节稳定与可切换的 T1B 细胞因子表达尚不清楚。了解此类机制将允许 我们在移植前对患者进行准确的风险分层，并阐明关键的监管机制 有助于个体患者的潜在免疫反应。我们假设转录 遗传或表观遗传机制的调节决定了 T1B 细胞因子比率、Breg 活性和 确定患者移植后的临床病程。该假设将通过检查遗传和 稳定与可切换 T1B 细胞因子表达的表观遗传基础，并通过确定临床是否 特征与这些变化相关。这些目标将使 Cherukuri 博士获得以下方面的专业知识：(i) 最先进的基因组和表观遗传分析方法（例如 ATACseq、亚硫酸氢盐测序、RNAseq）， (ii) 生物信息学，以及 (iii) 传统统计模型和无监督机器学习方法；在 除了磨练标准实验室技术之外。这些新的不可或缺的基础和转化方法 研究将有助于他过渡到从事有意义的独立研究的职业。这些技能将 在大学导师、顾问和合作者团队的指导下开发。匹兹堡。博士。 David Rothstein（1°导师）是一位知名的医师科学家，在移植免疫学方面拥有专业知识， 耐受性、Bregs 以及生物标志物研究。 Harinder Singh 博士（联合导师），国际 公认的科学家，在免疫反应的遗传和表观遗传调控方面具有专业知识，指导大学。 匹兹堡系统免疫学中心。两位导师都有出色的培训记录。咨询 由具有从基础移植免疫学到 转化和临床科学将每两年监测 Cherukuri 博士的进展。该提案将促进博士。 Cherukuri 的目标是作为一名拥有以下专业知识的终身医师科学家实现科学独立 Bregs，它们的转录调控和翻译应用作为生物标志物和治疗靶点。