BMP Signaling and Pulmonary Vasoreactivity

BMP 信号传导和肺血管反应性

• 批准号: 8307784
• 负责人: Mark P. de Caestecker
• 金额: $ 38.61万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-05 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8307784
• 关键词: Age; Agonist; BMPR2 gene; Binding; Blood Vessels; Cell Culture Techniques; Cell membrane; Cell surface; Chemicals; Chronic; Communities; Complex; Defect; Deletion Mutation; Development; Diagnosis; Disease; Endoplasmic Reticulum; Endothelial Cells; Exons; Foundations; Functional disorder; Genetic Models; Golgi Apparatus; Hypoxia; Inherited; Ligands; Lung; MAP Kinase Gene; MAPK14 gene; Mediating; Mediator of activation protein; Messenger RNA; Minor; Modeling; Molecular; Molecular Chaperones; Mus; Mutant Strains Mice; Mutation; Pathway interactions; Patients; Phenotype; Phosphorylation; Play; Process; Proteins; Proto-Oncogene Proteins c-akt; Pulmonary Hypertension; Regulation; Relative (related person); Role; Serotonin; Severities; Signal Pathway; Signal Transduction; Small Interfering RNA; Testing; Therapeutic; Transgenic Mice; Vascular Diseases; Vascular remodeling; bone morphogenetic protein receptor type II; insight; mRNA Decay; mutant; pressure; prevent; public health relevance; pulmonary arterial hypertension; receptor; response; trafficking

DESCRIPTION (provided by applicant): Patients with Familial Pulmonary Hypertension (FPAH) inherit heterozygous mutations in the BMP type 2 receptor BMPR2. It is unknown how these mutations cause pulmonary hypertension (PH). Half of these mutations escape Nonsense Mediated mRNA Decay and express BMPR2 mutant mRNA (NMD negative). Others are NMD positive and do not express mutant products. As NMD negative mutations express mutant gene products they may exert dominant inhibitory effects on the pulmonary vasculature. Recent evidence supports this: the age at diagnosis is younger in FPAH patients with NMD negative versus NMD positive BMPR2 mutations. This suggests NMD negative mutations cause more severe disease. Preliminary studies in mice carrying different Bmpr2 mutations suggest a mechanism by which this occurs. Mice with the heterozygous Bmpr2 Exon 2 deletion mutation (Bmpr2?Ex2/+) have more severe PH than mice carrying heterozygous null Bmpr2+/- mutations. This is associated with reduced eNOS activity and endothelial cell dysfunction in Bmpr2?Ex2/+ pulmonary vasculature. Over-expression of eNOS reverses the PH phenotype in Bmpr2?Ex2/+ mutant mice, indicating that eNOS deficiency plays a critical role in exacerbating PH. Furthermore, there is abnormal intracellular processing of the Bmpr2?Ex2 mutant product, and this interferes with eNOS function by abnormal intracellular sequestration of eNOS. As similar NMD negative Exon 2 deletion mutants are found in FPAH patients, these findings suggest that eNOS deficiency may be a common mechanism by which NMD negative BMPR2 mutations exacerbate PH in patients with FPAH. Three aims are proposed to test this hypothesis and define the mechanism by which this occurs. Aim 1 will compare vasoreactivity, PH responses and eNOS activity in the pulmonary vasculature of Bmpr2?Ex2/+ and Bmpr2+/- mice. Aims 2 will use cell culture techniques to evaluate how BMP signaling normally regulates eNOS expression and activity in pulmonary endothelial cells, while Aim 3 will compare eNOS regulation in Bmpr2+/- and Bmpr2?Ex2/+ mice, and will determine the molecular mechanism by which the Bmpr2?Ex2/+ mutant product exerts dominant inhibitory effects on eNOS activity in pulmonary endothelial cells. These studies will provide insight into how NMD positive FPAH BMPR2 mutations (like the Bmpr2+/- mutation) vs. NMD negative mutations (which are expressed and may behave like the Bmpr2?Ex2/+ mutation) modify vascular responses in patients with FPAH. By defining the mechanism by which Bmpr2?Ex2 inhibits eNOS they will also establish the foundation for therapeutic approaches to ameliorate PH by correcting NMD negative BMPR2 mutant processing defects in FPAH. PUBLIC HEALTH RELEVANCE: Pulmonary Arterial Hypertension is an invariably fatal disease for which there is no curative treatment. A major challenge for the scientific community therefore is to develop therapeutic approaches to prevent the progression of pulmonary vascular disease that develops in these patients. Our studies will provide important insight into a common pathway that regulates this process and identify approaches to interfere with this pathway and prevent the development of irreversible pulmonary vascular disease in these patients.

描述（由申请人提供）：家族性肺动脉高压（FPAH）患者遗传了BMP 2型受体BMPR 2的杂合突变。目前尚不清楚这些突变如何导致肺动脉高压（PH）。这些突变中的一半逃脱无义介导的mRNA衰变并表达BMPR 2突变体mRNA（NMD阴性）。其他人是NMD阳性，不表达突变产物。由于NMD阴性突变表达突变基因产物，它们可能对肺血管系统产生显性抑制作用。最近的证据支持这一点：NMD阴性与NMD阳性BMPR 2突变的FPAH患者的诊断年龄更年轻。这表明NMD阴性突变导致更严重的疾病。对携带不同Bmpr 2突变的小鼠的初步研究表明了这种情况发生的机制。Bmpr 2外显子2杂合子缺失突变小鼠（Bmpr 2？Ex 2/+）比携带杂合无效Bmpr 2 +/-突变的小鼠具有更严重的PH。这与Bmpr 2？Ex 2/+肺血管系统。eNOS的过度表达逆转了Bmpr 2？Ex 2/+突变小鼠，表明eNOS缺乏在加剧PH中起着关键作用。此外，有异常的细胞内处理Bmpr 2？Ex 2突变产物，并且这通过eNOS的异常细胞内隔离来干扰eNOS功能。由于在FPAH患者中发现了类似的NMD阴性外显子2缺失突变体，这些发现表明eNOS缺陷可能是NMD阴性BMPR 2突变加重FPAH患者PH的常见机制。提出了三个目标来检验这一假设，并确定发生这种情况的机制。目的1比较Bmpr 2？Ex 2/+和Bmpr 2 +/-小鼠。目标2将使用细胞培养技术来评估BMP信号如何正常调节肺内皮细胞中eNOS的表达和活性，而目标3将比较Bmpr 2 +/-和Bmpr 2中eNOS的调节。Ex 2/+小鼠，并将确定Bmpr 2？Ex 2/+突变产物对肺内皮细胞eNOS活性具有显性抑制作用。这些研究将深入了解NMD阳性FPAH BMPR 2突变（如Bmpr 2 +/-突变）与NMD阴性突变（表达并可能表现为Bmpr 2？Ex 2/+突变）改变FPAH患者的血管反应。通过定义Bmpr 2？Ex 2抑制eNOS，它们还将为通过纠正FPAH中的NMD阴性BMPR 2突变体加工缺陷来改善PH的治疗方法奠定基础。 公共卫生相关性：肺动脉高压是一种无法治愈的致命疾病。因此，科学界面临的一个主要挑战是开发治疗方法，以防止这些患者发生肺血管疾病的进展。我们的研究将提供重要的洞察到一个共同的途径，调节这一过程，并确定方法来干扰这一途径，并防止这些患者不可逆的肺血管疾病的发展。