Pre-Transplant Monocyte HDAC6 Expression and Risk of Primary Graft Dysfunction in Clinical Lung Transplant Recipients

临床肺移植受者移植前单核细胞 HDAC6 表达和原发性移植物功能障碍的风险

基本信息

批准号：
10527372
负责人：
Wayne William Hancock
金额：
$ 43.54万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-15 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10527372
关键词：
Acute Acute Lung Injury Adult Affect Binding Biological Markers Blood Cells Blood Vessels Blood specimen CD14 gene Cell surface Cells Characteristics Chronic lung disease Clinical Clinical Research Collaborations Complement Data Deacetylase Development Disease Disease susceptibility Engineering Enzyme-Linked Immunosorbent Assay Epigenetic Process Event Flow Cytometry Functional disorder Funding Gene Expression Genetic Transcription Genetic Variation HDAC6 gene Histones Hour Hypoxemia Immune Immunity Immunologics In Vitro Inflammatory Infrastructure Injury Ischemia Longterm Follow-up Lung Lung Transplantation Lung diseases Messenger RNA Modeling Morbidity - disease rate Mus Natural Immunity Operative Surgical Procedures Outcome Parents Pathogenesis Pathogenicity Pathway interactions Patients Peripheral Blood Mononuclear Cell Phenotype Population Postoperative Period Process Production Proteins Publishing Pulmonary Edema Regulation Reperfusion Injury Research Research Personnel Risk Risk Factors Role Sample Size Source TLR4 gene Testing Therapeutic Training Transplant Recipients Transplantation Ubiquitin United States National Institutes of Health Up-Regulation Zinc Fingers adaptive immunity biomarker validation cytokine genomic locus graft dysfunction in vivo individual variation inhibitor inhibitor therapy lung ischemia monocyte mortality mouse model novel novel diagnostics novel therapeutics overexpression participant enrollment pathogen predicting response prognostic response small molecule inhibitor transcriptome transcriptome sequencing translational study treatment response

项目摘要

Project Summary Primary Graft Dysfunction (PGD) remains a leading cause of early morbidity and mortality in lung transplant (Tx) recipients. The Lung Transplant Outcomes Group (LTOG) has been studying this problem for the past decade and a newly NIH-funded LTOG-related project (1U01HL14535-01), headed by Dr. Jason Christie (UPENN), involves long-term follow- up of adult lung transplant recipients. In preliminary studies, undertaken in collaboration with LTOG, we found that patients who subsequently developed PGD had significantly elevated pre-Tx levels of HDAC6 mRNA in their peripheral blood mononuclear cells. We identified CD14+ monocytes as the main source of upregulated HDAC6 in pre-Tx PGD-prone patients, observed an inflammatory phenotype of those monocytes after stimulation in vitro, and found that HDAC6 targeting significantly decreased cytokine production in an in vivo murine lung model of ischemia/reperfusion injury. Accordingly, we hypothesize that recipient blood monocytes with upregulated HDAC6 expression pre-lung Tx are important drivers of graft injury and development of PGD, and that such elevated HDAC6 expression in monocytes may be of prognostic and therapeutic significance. We propose to test this hypothesis by studying pre-Tx blood samples from patients enrolled in the parent U01 that began June 15, 2019. Aim 1: Determine if the phenotype of upregulated HDAC6 in monocytes of patients listed for lung Tx is a risk factor for developing PGD? We will expand upon our preliminary data using a larger sample size to evaluate 1.1) pre-Tx levels of monocyte HDAC6 expression, including mRNA and protein levels, and co-expression of classical markers of monocyte activation; and 1.2) post-Tx events (PGD with grade) in relation to the pre-Tx monocyte phenotype. Aim 2: What are the mechanisms by which HDAC6 alters key monocyte characteristics? We will: 2.1) evaluate cytokine expression (qPCR, flow cytometry, ELISA) by monocytes isolated from pre-Tx recipients and healthy donors, in relation to their HDAC6 levels; and 2.2) study how high HDAC6 expression regulates TLR/MyD88 pathway activation. Aim 3: Test whether HDAC6 inhibitor (HDAC6i) therapy can reverse or diminish the effects of monocyte dysfunction, including ex vivo and in a murine model of PGD. We will: 3.1) test effects of HDAC6i on monocyte activation, cytokine production and interaction with other immune cells; 3.2) test effects of one or more novel HDAC6i small molecules (monocyte specific HDAC6i, HDAC6 zinc-finger ubiquitin binding domain inhibitor and HDAC6i PROTAC); and 3.3) apply HDAC6i in vivo in a recently described new murine model of PGD. Importantly, our studies will • delineate pathogenic mechanisms of disease; • identify mechanisms or factors that influence and/or predict response to treatment; and • discover or validate biomarkers of disease development and/or progression.

项目概要原发性移植物功能障碍 (PGD) 仍然是肺移植 (Tx) 早期发病和死亡的主要原因肺移植结果小组（LTOG）在过去十年中一直在研究这个问题，并且最近提出了一项新的研究。由 Jason Christie 博士 (UPENN) 领导的 NIH 资助的 LTOG 相关项目 (1U01HL14535-01) 涉及长期跟踪在与 LTOG 合作进行的初步研究中，我们发现患者随后进行 PGD 的患者外周血中 HDAC6 mRNA 的 Tx 前水平显着升高我们确定 CD14+ 单核细胞是 Tx 前 PGD 倾向中 HDAC6 上调的主要来源。患者，观察这些单核细胞在体外刺激后的炎症表型，发现 HDAC6 在缺血/再灌注损伤的体内小鼠肺模型中，靶向显着减少细胞因子的产生。因此，我们发现肺 Tx 前 HDAC6 表达上调的受体血单核细胞是移植物损伤和 PGD 发展的重要驱动因素，单核细胞中 HDAC6 表达升高可能与我们建议通过研究 Tx 前的血液样本来检验这一假设。 2019 年 6 月 15 日开始入组母体 U01 的患者。目标 1：确定 HDAC6 的表型是否上调接受肺 Tx 治疗的患者的单核细胞中的 Tx 是发生 PGD 的危险因素吗？使用较大样本量的数据来评估1.1）单核细胞HDAC6表达的pre-Tx水平，包括mRNA和蛋白质水平和单核细胞激活经典标志物的共表达；以及 1.2) Tx 后事件（PGD 与等级）的关系目标 2：HDAC6 改变关键单核细胞的机制是什么。我们将： 2.1) 评估分离自单核细胞的细胞因子表达（qPCR、流式细胞术、ELISA） Tx 前受者和健康捐献者的 HDAC6 水平；2.2) 研究 HDAC6 表达有多高；调节 TLR/MyD88 通路激活目标 3：测试 HDAC6 抑制剂 (HDAC6i) 治疗是否可以逆转或逆转。减少单核细胞功能障碍的影响，包括离体和 PGD 小鼠模型。我们将： 3.1) 测试。 HDAC6i对单核细胞活化、细胞因子产生以及与其他免疫细胞相互作用的影响；3.2）测试效果；一种或多种新型 HDAC6i 小分子（单核细胞特异性 HDAC6i、HDAC6 锌指泛素结合域 3.3) 在最近描述的新的 PGD 小鼠模型中应用体内 HDAC6i。重要的是，我们的研究将 • 阐明疾病的致病机制 • 确定影响的机制或因素；和/或预测治疗反应；以及 • 发现或验证疾病发生和/或进展的生物标志物。