Role of F-Box Proteins in Lung Transplantation

F-Box 蛋白在肺移植中的作用

• 批准号: 9382219
• 负责人: Beibei Chen
• 金额: $ 53.68万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-15 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9382219
• 关键词: Acute; Adaptor Signaling Protein; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Attenuated; Binding; Biology; Blood; Bronchiolitis Obliterans; Bronchoalveolar Lavage; CD28 gene; Calcineurin inhibitor; Calcium-Binding Proteins; Calmodulin; Cells; Chronic; Cyclosporine; Data; Development; Disease; Drug Targeting; Employee Strikes; Equilibrium; F-Box Proteins; FBXL2 gene; Foundations; Functional disorder; Future; Graft Rejection; Human; Immune response; Immune system; Immunity; Immunobiology; Immunology; Immunosuppression; In Vitro; Inflammation; Inflammatory; Interferon Type II; Laboratories; Lead; Life; Lung; Lung Transplantation; Lung diseases; Mediating; Messenger RNA; Modeling; Molecular; Mus; Organ Transplantation; Outcome; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Play; Procedures; Production; Proteins; Registries; Regulation; Research Personnel; Risk Factors; Role; Sampling; Savings; Science; Severities; Site; Solid; Syndrome; T cell response; T-Cell Activation; T-Lymphocyte; T-bet protein; TNF Receptor-Associated Factors; TNF gene; Testing; Therapeutic; Therapeutic immunosuppression; Translating; Translational Research; Transplant Recipients; Transplantation; Transplanted tissue; Treatment Efficacy; Ubiquitin; Ubiquitination; Universities; Up-Regulation; allograft rejection; base; biobank; cytokine; drug development; improved; improved outcome; inhibitor/antagonist; isoimmunity; lung allograft; multidisciplinary; novel; novel drug class; novel strategies; novel therapeutics; pre-clinical; prevent; programs; response; small molecule; small molecule inhibitor; success; targeted treatment; transcription factor; ubiquitin-protein ligase

Abstract: Lung transplantation (LT) is the only therapeutic option for patients with end-stage lung disease; however, survival lags behind other organ transplants, and is primarily due to chronic lung allograft dysfunction (CLAD), most often the bronchiolitis obliterans syndrome (BOS). Episodes of acute cellular rejection (ACR) are common and the major risk factor for CLAD, despite calcineurin inhibitor-based (e.g., cyclosporine; CSA) conventional immunosuppression therapy (IST). Experimental transplant models and humans studies indicate that Type-1 T cell immunity, marked by IFN-γ production, and driven by the transcription factor, T-bet, play a key role in allograft rejection. Thus, new and effective therapeutic strategies represent a major unmet need in LT and are desperately needed to prevent lung rejection and improve outcomes. We have discovered a novel, pro-inflammatory ubiquitin, E3 ligase subunit, Fbxo3, which degrades an anti-inflammatory calmodulin (CaM)- associated protein called Fbxl2. Our preliminary data show Fbxo3 is induced during ACR in the mouse orthotopic lung transplant (mouse OLT) model, along with T-bet, while Fbxl2 is markedly reduced. Further, using a first-in-class novel small molecule inhibitor of Fbxo3, BC-1261, reduces T-bet/Type-1 immunity and ACR severity, and preserves Fbxl2 in our early studies. Our studies in lung transplant recipients (LTRs) show a predominance of Type-1 alloimmune responses in the lung and blood, up-regulation of Fbxo3, and inhibition of immune responses with BC-1261. Therefore, we hypothesize that Fbxl2 is a critical regulator of T-bet/Type-1 immunity in T cells, and that Fbxo3: Fbxl2 balance and T-bet/IFN-γ induction are key determinants of lung allograft rejection versus acceptance. To test this, we propose 3 aims. In Aim 1, we will test the hypothesis that induction of Fbxo3 degrades Fbxl2 and increases T-bet in activated T cells. We will determine whether Fbxl2 ubiquitinates T-bet, whether CaM acts as a linker molecule in regulating T-bet, and test the regulation of Fbxo3 during T cell activation. In Aim 2, using the mouse OLT model, we will test the hypothesis that the balance of functional Fbxo3: Fbxl2 and T-bet/Type-1 immunity are key determinants of lung allograft outcomes. Here, we will assess our novel Fbxo3 inhibitor, BC-1261 versus CSA on ACR or obliterative airways disease (OAD; modeling BOS), and test whether lung rejection is reduced in novel Fbxo3-deficient mice. In Aim 3, we will test the hypothesis that the Fbxo3 and T-bet/Type-1 immunity pathways are putative targets in human LT and whether BC-1261 inhibits alloimmune responses. Making use of our expanding LTR registry/biorepository of bronchoalveolar lavage (BAL) and PBMC samples, we will determine whether F-box proteins play an important role in lung allograft and systemic alloimmunity. The PIs, Drs. McDyer and Chen, bring multidisciplinary expertise combining ubiquitin biology, mouse and human lung transplant immunology, and will use the multiple PI format. Success in this R01 will be transformative in the science to prevent lung allograft rejection, and lay the foundation to test a first-in-class therapy targeting the pro-inflammatory ubiquitin, E3 ligase subunit, Fbxo3.

摘要：肺移植（LT）是末期肺部疾病患者的唯一治疗选择； 但是，生存滞后在其他器官移植后面，主要是由于慢性肺同种异体功能障碍 （外壳），最常见的是细支气管炎闭塞综合征（BOS）。急性细胞排斥（ACR）的发作是 clad的常见和主要危险因素，基于钙调蛋白抑制剂（例如环孢素; CSA） 常规免疫抑制疗法（IST）。实验移植模型和人类研究表明 由IFN-γ产生标记的那种1型T细胞免疫，由转录因子T-BET驱动，发挥作用 同种异体拒绝中的关键作用。那是新的有效的治疗策略代表了主要的未满足需求 LT并且迫切需要防止肺部排斥和改善预后。我们发现了一部小说， 促炎性泛素，E3连接酶亚基，FBXO3，降解抗炎钙调蛋白（CAM） - 相关蛋白称为FBXL2。我们的初步数据表明，在ACR中，在鼠标中诱导了FBXO3 原位肺移植（小鼠OLT）模型以及T-bet，而FBXL2显着降低。更远， 使用FBXO3的第一类新型小分子抑制剂，BC-1261，降低T-Bet/type-1免疫抑制剂和 ACR严重性，并在我们的早期研究中保留FBXL2。我们对肺移植受者（LTR）的研究表明 肺和血液中1型同种免疫反应的优势，FBXO3的上调以及抑制 BC-1261的免疫反应。因此，我们假设FBXL2是T-Bet/Type-1的关键调节剂 T细胞的免疫力，FBXO3：FBXL2平衡和T-BET/IFN-γ诱导是肺的关键决定剂 同种异体移植拒绝与接受。为了测试这一点，我们提出了3个目标。在AIM 1中，我们将检验以下假设 FBXO3的诱导降低了FBXL2并增加了活化的T细胞中T-BET。我们将确定FBXL2是否 泛素T-bet，CAM是否充当调节T-BET中的接头分子，并测试FBXO3的调节 在T细胞激活期间。在AIM 2中，使用鼠标OLT模型，我们将检验以下假设。 功能性FBXO3：FBXL2和T-BET/Type-1免疫历史是肺同种异体结局的关键决定者。在这里，我们 将评估我们新颖的FBXO3抑制剂，BC-1261与CSA在ACR或闭塞气道疾病上（OAD; 对BOS进行建模），并测试在新型FBXO3缺陷小鼠中是否减少了肺排斥。在AIM 3中，我们将测试 FBXO3和T-BET/Type-1免疫病是人类LT和 BC-1261是否抑制同种免疫反应。利用我们扩展的LTR注册表/生物座 支气管肺泡灌洗（BAL）和PBMC样品，我们将确定F-box蛋白是否发挥重要作用 在肺同类和系统性同种免疫中的作用。 pis，博士。 McDyer和Chen，带来多学科 结合泛素生物学，小鼠和人肺移植免疫学的专业知识，将使用多种 PI格式。在此R01中的成功将在科学中具有变革性，以防止肺同种异体移植排斥，并放置 测试针对促炎性泛素E3连接酶亚基FBXO3的第一类治疗的基础。