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

• 批准号: 10152233
• 负责人: Wayne William Hancock
• 金额: $ 45.44万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 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/10152233
• 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; Clinical; Clinical Research; Collaborations; Complement; Data; Deacetylase; Development; Disease; Disease susceptibility; Engineering; Enrollment; 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; 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; cytokine; genomic locus; in vivo; individual variation; inhibitor/antagonist; lung ischemia; monocyte; mortality; mouse model; novel; novel diagnostics; novel therapeutics; overexpression; pathogen; predicting response; prognostic; response; small molecule inhibitor; transcriptome; transcriptome sequencing; translational study; treatment response

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.

项目摘要 原发性移植功能障碍（PGD）仍然是肺移植早期发病和死亡率的主要原因（TX） 收件人。在过去的十年中，肺移植成果组（LTOG）一直在研究这个问题和新 由NIH资助的LTOG相关项目（1U01HL14535-01），由Jason Christie博士（UPENN）领导，涉及长期跟进 - 成年肺移植受者。在与LTOG合作进行的初步研究中，我们发现患者 后来出现PGD的人在其外周血中的HDAC6 mRNA水平显着升高 单核细胞。我们将CD14+单核细胞确定为PRE-TX PGD易于更新的HDAC6的主要来源 患者观察到体外刺激后这些单核细胞的炎症表型，发现HDAC6 在缺血/再灌注损伤的体内鼠肺模型中，靶向可显着降低细胞因子的产生。 据此，我们假设接受带有更新的HDAC6表达前肺前TX的受体血液单核细胞为 嫁接损伤和PGD发育的重要驱动因素，单核细胞中这种升高的HDAC6表达可能是 预后和治疗意义的意义。我们建议通过研究来自 入学的患者参加了2019年6月15日开始的父级U01。目标1：确定更新的HDAC6表型是否是否 在列出的肺部单核细胞中，TX是发展PGD的危险因素吗？我们将扩展我们的初步 使用较大样本量评估1.1）单核细胞HDAC6表达的数据，包括mRNA和蛋白质 单核细胞激活的经典标记的水平和共表达；和1.2）TX后事件（具有等级的PGD） 到TX前单核细胞表型。目标2：HDAC6改变关键单核细胞的机制是什么 特征？我们将：2.1）通过从从中分离的单核细胞评估细胞因子表达（qPCR，流式细胞仪，ELISA） 与HDAC6水平相关的TX前接受者和健康的捐助者； 2.2）研究HDAC6表达高的高度 调节TLR/MYD88途径激活。目标3：测试HDAC6抑制剂（HDAC6I）疗法是否可以逆转或 减少单核细胞功能障碍的影响，包括离体和PGD的鼠模型。我们将：3.1）测试 HDAC6I对单核细胞激活，细胞因子产生以及与其他免疫小球的相互作用的影响； 3.2）测试效果 一个或多个新型HDAC6I小分子（单核细胞特异性HDAC6I，HDAC6锌指泛素结合结构域 抑制剂和HDAC6I PROTAC）； 3.3）在最近描述的PGD新鼠模型中应用HDAC6I在体内应用。 重要的是，我们的研究将•描述疾病的致病机制； •确定影响的机制或因素 和/或预测对治疗的反应； •发现或验证疾病发展和/或进展的生物标志物。