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 尽管短期结局显着，但后期肾脏同种异体损失的速度却没有提高。这是 越来越多地归因于持续存在的微妙免疫损伤的累积影响 未被发现。临床参数，甚至监视活检的预测价值有限，强调 需要新的预测生物标志物。 Cherukuri博士表明：（i）人类的布雷格活动最好的特征是 未成熟的T1过渡B细胞（T1B）将IL-10/TNFα表达的比率； （ii）此T1B IL-10/TNFα比 3 MOS移植物预测随后的急性排斥（AR），同种异体移植纤维化，EGFR和移植物减少 生存。此外，在移植之前对T1B IL-10/TNFα比率的评估表明，〜75％ 患者，移植后的细胞因子比保持稳定，并预测其AR的风险和随后的风险 结果（预定）。相反，在约25％的患者中，细胞因子比从高或低转移 及其随后的结果的风险与轮班的方向相同。这些机制 调节稳定与可切换的T1B细胞因子表达尚不清楚。了解这种机制将允许 我们要在移植之前准确地划分患者，并阐明关键的调节机制 有助于个体患者的潜在免疫反应性。我们假设该转录 通过遗传或表观遗传机制调节决定了T1B细胞因子比率，Breg活性和 确定患者移植后临床课程。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，并通过确定是否临床 特征与这些变化有关。这些目标将使Cherukuri博士获得以下方面的专业知识：（i） ART先进的基因组和表观遗传分析方法（例如Atacseq，Bisulfite测序，RNASEQ），RNASEQ）， （ii）生物信息学和（iii）传统的统计建模以及无监督的机器学习方法； 补充标准实验室技术。这些新的基本和翻译必不可少的方法 研究将促进他过渡到执行有意义的独立研究的职业。这些技能将是 在大学的导师，顾问和合作者团队的指导下开发。匹兹堡。博士 大卫·罗斯斯坦（David Rothstein）（1°精神）是一位既定的身体科学家，具有移植免疫学专业知识， 耐受性和布雷格以及生物标志物研究。国际上的Harinder Singh博士（Co-Entor） 公认的科学家在免疫反应的遗传和表观遗传调节方面具有专业知识，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 专家委员会的专家委员会从基本移植免疫学到 翻译和临床科学将每两年监测Cherukuri博士的进步。该建议将促进博士。 Cherukuri的目标是将科学独立作为具有专业知识的统治曲目的身体科学家 Bregs，其转录调节和翻译应用为生物标志物，并作为治疗靶标。