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.

翻译后摘要：肺移植（LT）是终末期肺病患者的唯一治疗选择; 然而，存活率落后于其他器官移植，主要是由于慢性肺移植功能障碍 （CLAD），最常见的是闭塞性细支气管炎综合征（BOS）。急性细胞排斥反应（ACR）的发生率是 CLAD的常见和主要风险因素，尽管基于钙调神经磷酸酶（例如，环孢菌素 常规免疫抑制疗法（IST）。实验移植模型和人体研究表明， 1型T细胞免疫，以IFN-γ的产生为标志，并由转录因子T-bet驱动， 在同种异体移植排斥反应中起关键作用。因此，新的有效治疗策略代表了一个未满足的主要需求 为了防止肺排斥反应和改善预后，迫切需要LT和LT。我们发现了一本小说， 促炎泛素，E3连接酶亚基，Fbxo 3，其降解抗炎钙调蛋白（CaM）。 一种叫做Fbxl 2的蛋白。我们的初步数据显示，Fbxo 3在小鼠ACR过程中被诱导， 在原位肺移植（小鼠奥尔特）模型中，沿着T-bet，而Fbxl 2显著降低。此外，本发明还 使用一流的新型Fbxo 3小分子抑制剂BC-1261，降低T-bet/1型免疫力， ACR严重程度，并在我们的早期研究中保留Fbxl 2。我们在肺移植受者（LTR）中的研究显示， 肺和血液中1型同种免疫应答的优势，Fbxo 3的上调， BC-1261的免疫应答。因此，我们假设Fbxl 2是T-bet/Type-1的关键调节因子。 T细胞中免疫性、FbxO 3：FbxL 2平衡和T-bet/IFN-γ诱导是肺的关键决定因素 同种异体移植排斥与接受。为了验证这一点，我们提出了三个目标。在目标1中，我们将检验以下假设： FbxO 3的诱导降解FbxL 2并增加活化T细胞中的T-bet。我们将确定Fbxl 2是否 泛素化T-bet，CaM是否作为连接分子调节T-bet，并测试Fbxo 3的调节 在T细胞活化过程中。在目标2中，使用小鼠奥尔特模型，我们将测试以下假设： 功能性FbxO 3：FbxL 2和T-bet/1型免疫是肺同种异体移植物结果的关键决定因素。这里我们 将评估我们的新型Fbxo 3抑制剂BC-1261与CSA对ACR或闭塞性气道疾病（OAD; 建模BOS），并测试肺排斥反应是否在新的FbxO 3缺陷小鼠中减少。在目标3中，我们将测试 假设Fbxo 3和T-bet/1型免疫途径是人LT中的假定靶点， BC-1261是否抑制同种免疫应答。利用我们不断扩大的LTR注册/生物储存库， 支气管肺泡灌洗（BAL）和PBMC样本，我们将确定F-盒蛋白是否发挥重要作用， 在肺同种异体移植和全身同种免疫中的作用。PI，McDyer博士和Chen博士，带来了多学科 专业知识结合泛素生物学，小鼠和人类肺移植免疫学，并将使用多种 PI格式。R 01的成功将在预防肺同种异体移植排斥反应的科学方面具有革命性意义， 这是测试针对促炎性泛素E3连接酶亚基Fbxo 3的一流疗法的基础。