Proteomics and Gene Expression in Hereditary Disorders of Connective Tissue

结缔组织遗传性疾病的蛋白质组学和基因表达

• 批准号: 8335950
• 负责人: Nazli Mcdonnell
• 金额: $ 16.25万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8335950
• 关键词: Affect; Aneurysm; Angiotensin Receptor; Arteries; Blood Vessels; Body Fluids; Cell Surface Receptors; Connective Tissue; Disease; Ehlers-Danlos Syndrome; Electrophoresis; Extracellular Matrix Proteins; FBN1; Family member; Fibroblasts; Fibromuscular Dysplasia; Gene Expression; Gene Mutation; Genes; Genome; Goals; Hereditary Disease; Immunochemistry; Lead; Loeys-Dietz Syndrome; Losartan; Marfan Syndrome; Mass Spectrum Analysis; Methodology; Molecular Profiling; Mutation; Pathway interactions; Patients; Phenotype; Proteins; Proteomics; Protocols documentation; Publications; Publishing; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Stickler syndrome; Syndrome; Tissues; Transforming Growth Factor beta; Western Blotting; protein profiling

Hereditary disorders of connective tissue, such as Ehlers Danlos (EDS), Loeys-Dietz, Marfan, Stickler, Fibromuscular Dysplasia (FMD), Vascular EDS (VEDS), and Familial Aneurysm syndromes are genetically and clinically heterogeneous. While many of the causative genes are known, many others are not. Many affected family members may have mutations in unique genes that may be impractical to identify with usual methodologies. The goal of this project is to develop a comprehensive gene expression and a complementary protein profiling approach utilizing samples collected from affected patients through an IRP-approved protocol (2003-086). The hypothesis is that conditions that share a similar phenotype in terms of complications, regardless of the underlying gene mutation, the expression and proteomic profiles wil turn out to similar and amenable to treatment utilizing the same targets. For example, Loeys-Dietz syndrome is caused by mutations in TGFbetaR1&2, which are cell surface receptors, and Marfan syndrome is caused by mutations in FBN1, a structural extracellular matrix protein. Despite different causative mutations, both syndromes lead to derangements of the TGFbeta pathway, have phenotypic similarities, and are amenable to treatment by losartan, which is an angiotensin receptor blocker that is also a modulator of TGFbeta expression. We are utilizing whole genome expression arrays, RT-PCR 2-D protein electrophoresis, mass spectroscopy, western blot and immunochemistry approaches to investigate the consequences of known mutations in VEDS patients, and comparing the profiles of patients without such mutations but with similar phenotypic features, such as FMD. Recent results indicate that circulating TGFbeta1 levels are increased in patients with Marfan syndrome, VEDS, and FMD. The results of our findings in Marfan syndrome have been published and the other disorders are being prepared for publication. This is envisioned as a long term project given that more than 300 unique patient fibroblast lines from connective tissue patients are being studied in the lab.

结缔组织的遗传性疾病，如Ehlers Danlos（EDS）、Loeys-Dietz、Marfan、Stickler、纤维肌性发育不良（FMD）、血管EDS（VEDS）和家族性动脉瘤综合征在遗传和临床上是异质性的。虽然许多致病基因是已知的，但许多其他基因还不知道。许多受影响的家庭成员可能有独特的基因突变，这可能是不切实际的，以确定与通常的方法。 该项目的目标是开发一个全面的基因表达和互补的蛋白质分析方法，利用通过IRP批准的方案（2003-086）从受影响的患者收集的样本。该假设是，在并发症方面具有相似表型的病症，无论潜在的基因突变如何，表达和蛋白质组谱将变得相似并且适于利用相同靶标进行治疗。例如，Loeys-Dietz综合征是由细胞表面受体TGF β R1和2突变引起的，而Marfan综合征是由FBN 1突变引起的，FBN 1是一种结构性细胞外基质蛋白。尽管致病突变不同，但两种综合征均导致TGF β通路紊乱，具有表型相似性，并可接受氯沙坦治疗，氯沙坦是一种血管紧张素受体阻滞剂，也是TGF β表达的调节剂。 我们正在利用全基因组表达阵列、RT-PCR 2-D蛋白电泳、质谱、蛋白质印迹和免疫化学方法来研究VEDS患者中已知突变的后果，并比较没有此类突变但具有相似表型特征（如FMD）的患者的概况。最近的研究结果表明，循环TGF β 1水平在马凡综合征，VEDS和FMD患者中升高。 我们在马凡氏综合征的研究结果已经发表，其他疾病的研究结果正在准备发表。 这被设想为一个长期项目，因为实验室正在研究来自结缔组织患者的300多个独特的患者成纤维细胞系。