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: 10542385
• 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/10542385
• 关键词: ATAC-seq; Acute; Adult; Advisory Committees; Affect; Allografting; Autoimmune; Autoimmune Diseases; B-Cell Acute Lymphoblastic Leukemia; 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 response; Immunologics; Immunology; Immunosuppression; Individual; Inflammation; Injury; Interleukin-10; International; Kidney Transplantation; Lead; Maps; Mentors; Methylation; 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.

项目摘要/摘要 尽管短期效果显著，但晚期肾移植失败率并未改善。这是 越来越多地归因于持续的微妙免疫损伤的累积效应，这种损伤往往 未被发现。临床参数甚至监视活检的预测价值有限，突显出 需要新的预测生物标记物。Cherukuri博士表明：(I)人类的Breg活动最好的特征是 未成熟T1移行B细胞(T1B)表达IL-10/肿瘤坏死因子α的比率 移植后3个月，预测随后的急性排斥反应(AR)、移植肾纤维化、EGFR降低和移植物 生死存亡。此外，移植前对T1B IL-10/肿瘤坏死因子α比率的评估显示，在~75%的 患者，移植后细胞因子比率保持稳定，并预测他们患AR和随后的风险 结果(预先确定)。相比之下，在约25%的患者中，细胞因子比率从高到低或从低到低 到了很高的水平，他们对后续结果的风险随着转变的方向而变化。这些机制可以 调节稳定的和可切换的T1B细胞因子的表达是未知的。了解这些机制将使 美国在移植前准确地对患者进行风险分层，并阐明 有助于个体患者潜在的免疫反应性。我们假设转录的 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。这一假说将通过检查基因和 稳定的与可转换的T1B细胞因子表达的表观遗传学基础，并通过确定临床上 特征与这些变化相关。这些目标将使Cherukuri博士能够获得以下方面的专门知识：(I) 本领域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)， (2)生物信息学；(3)传统统计建模和无监督机器学习方法； 除了磨练标准的实验室技术之外。这些新的必不可少的基本和翻译方法 研究将有助于他过渡到从事有意义的独立研究的职业。这些技能将是 在大学导师、顾问和合作者团队的指导下开发。匹兹堡的。Dr。 大卫·罗斯坦(1°Mentor)是一位在移植免疫学方面拥有专业知识的知名内科科学家， 耐受性、Bregs以及生物标记物研究。哈林德·辛格博士(共同导师)，国际专家 著名科学家，在免疫反应的遗传和表观遗传调控方面具有专门知识，是该大学的负责人。 匹兹堡系统免疫学中心的。这两位导师都有出色的培训记录。忠告 由经验丰富的研究人员组成的委员会，其专业知识从基础移植免疫学到 翻译和临床科学将每两年监测一次Cherukuri博士的进展。这项提议将提拔Dr。 切尔库里的目标是作为一名终身制医生-科学家实现科学独立 Bregs，它们作为生物标志物和治疗靶点的转录调控和翻译应用。