Molecular regulation of BMPRII stability in lung fibrosis

肺纤维化中 BMPRII 稳定性的分子调节

• 批准号: 10712273
• 负责人: Yutong Zhao
• 金额: $ 50.02万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712273
• 关键词: Ablation; Aging; Architecture; BMP4; BMPR2 gene; Back; Bone Morphogenetic Proteins; Cells; Chronic; Cicatrix; Data; Development; Docking; Down-Regulation; Epithelial Cells; Exhibits; Extracellular Matrix; Failure; Fibroblasts; Fibrosis; Genetic; Impairment; Injury; Knowledge; Link; Lipid Peroxidation; Lung; Lung diseases; Lysosomes; Mediating; Modeling; Molecular; Mus; Myofibroblast; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Play; Polyubiquitin; Polyubiquitination; Production; Property; Proteins; Pulmonary Fibrosis; Regulation; Resolution; Role; Scheme; Signal Induction; Signal Pathway; Signal Transduction; Signaling Protein; Site; System; Testing; Therapeutic; Transforming Growth Factors; Ubiquitination; Up-Regulation; bone morphogenetic protein receptors; cell injury; effective therapy; fibrotic lung; fibrotic lung disease; gain of function; healing; idiopathic pulmonary fibrosis; loss of function; lung development; novel; novel therapeutic intervention; overexpression; prevent; protein function; proteostasis; repaired; response; restoration; success; therapy development; ubiquitin-protein ligase

Abstract Idiopathic Pulmonary Fibrosis (IPF) is a chronic, irreversible, aging-associated, and ultimately fatal lung disease. The median survival of pulmonary fibrosis patients is only 4-5 years. There is no treatment to reverse fibrosis and cure IPF. Development of anti-fibrotic therapeutics is an unmet need in the treatment of IPF. The relentless progression of IPF is due in part to the failure of fibrosis resolution. The development of therapies for IPF relies on the comprehensive understanding of fibrosis resolution pathways. Accumulating evidence shows that activation of BMPs signaling induces myofibroblast de-differentiation and fibrosis resolution. A key receptor, BMP receptor II (BMPRII), in BMPs signaling has been shown to be reduced in fibrotic lungs; thus, restoration of BMPRII in fibrotic lungs is a potential therapy to treat IPF. However, molecular regulation of BMPRII stability has not been well studied. In our preliminary data, we discovered that (i) BMPRII is degraded in the lysosome system in response to TGF-β1 and lipid peroxidation inducers, which play critical roles in the development of lung fibrosis; (ii) Nedd4L stabilizes BMPRII; (iii) downregulation of Nedd4L reduced BMP4 signaling and the effects were rescued by overexpression of BMPRII; (iv) overexpression of Nedd4L promoted de-differentiation of lung myofibroblasts. Based on these novel observations, we hypothesized that Nedd4L promotes BMPRII stability and facilitates BMPs/BMPRII-mediated myofibroblast de-differentiation and pulmonary fibrosis resolution. We propose three Specific Aims to evaluate our hypothesis. First, we will determine the regulatory mechanisms by which Nedd4L stabilizes BMPRII. We will identify the ubiquitination and Nedd4L docking sites on BMPRII and determine the effect of Nedd4L-mediated K63-linked ubiquitination on BMPRII internalization and stability. Next, we will determine if Nedd4L suppression is essential for TGF-β1- and lipid peroxidation-induced BMPRII degradation. Finally, we will determine if Nedd4L facilitates lung fibrosis resolution through stabilization of BMPRII and promotion of BMPs-mediated myofibroblast de-differentiation and inactivation. We will determine if elevated BMPRII stability by Nedd4L facilitates myofibroblast de-differentiation in ex-vivo cultured lung myofibroblasts. An inducible fibroblast specific Nedd4L depletion mouse will be used in both resolving and non- resolving lung fibrosis models. The proposed studies will address key knowledge gaps regarding molecular regulation of BMPRII stability and anti-fibrotic effects of BMPs/BMPRII. Success of the proposed studies will suggest that targeting Nedd4L/BMPRII to rescue BMPRII expression in fibrotic lungs may lead to new opportunities to halt pro-fibrotic progression and promote fibrosis resolution.

摘要 特发性肺纤维化（IPF）是一种慢性、不可逆、衰老相关的最终致死性肺部疾病。 肺纤维化患者的中位生存期仅为4-5年。没有治疗方法可以逆转纤维化 治愈IPF。开发抗纤维化治疗药物是IPF治疗中尚未满足的需求。无情的 IPF的进展部分归因于纤维化消退的失败。IPF治疗的开发依赖于 对纤维化解决途径的全面理解。越来越多的证据表明， BMP信号传导的激活诱导肌成纤维细胞去分化和纤维化消退。一种关键受体，骨形态发生蛋白 已显示在纤维化肺中BMP信号传导中的BMPRII受体（BMPRII）减少;因此， 纤维化肺中的BMPRII是治疗IPF的潜在疗法。然而，BMPRII稳定性的分子调控具有 没有得到很好的研究。在我们的初步数据中，我们发现（i）BMPRII在溶酶体系统中被降解， 对TGF-β1和脂质过氧化诱导剂的反应，这在肺纤维化的发展中起关键作用; (ii)Nedd 4L稳定BMPRII;（iii）下调Nedd 4L减少BMP 4信号传导，其作用是： （iv）Nedd 4L的过表达促进肺去分化 肌成纤维细胞基于这些新的观察，我们假设Nedd 4L促进BMPRII稳定性， 并促进BMP/BMPIII介导的肌成纤维细胞去分化和肺纤维化消退。我们 提出三个具体目标来评估我们的假设。第一，我们将确定监管机制， 其中Nedd 4L稳定BMPRII。我们将鉴定BMPRII上的泛素化和Nedd 4L对接位点， 确定Nedd 4L介导的K63连接的泛素化对BMPRII内化和稳定性的影响。接下来， 我们将确定Nedd 4L抑制是否是TGF-β1和脂质过氧化诱导的BMPRII所必需的 降解最后，我们将确定Nedd 4L是否通过稳定 BMPRII和促进BMP介导的肌成纤维细胞去分化和失活。我们将确定是否 通过Nedd 4L提高的BMPRII稳定性促进离体培养肺中的肌成纤维细胞去分化 肌成纤维细胞诱导型成纤维细胞特异性Nedd 4L耗竭小鼠将用于消退和非消退。 解析肺纤维化模型。拟议的研究将解决有关分子生物学的关键知识差距， BMPRII稳定性的调节和BMP/BMPRII的抗纤维化作用。拟议研究的成功将 提示靶向Nedd 4L/BMPRII以拯救纤维化肺中BMPRII表达可能导致新的 阻止促纤维化进展和促进纤维化消退的机会。