Proteomics and Gene Expression in Hereditary Disorders of Connective Tissue

遗传性结缔组织疾病，如Ehler Danlos(EDS)、Loeys-Dietz、Marfan、Stickler、纤维肌肉发育不良(FMD)、血管EDS(VEDS)和家族性动脉瘤综合征在遗传和临床上是不同的。虽然许多致病基因是已知的，但其他许多基因却是未知的。许多受影响的家庭成员可能在独特的基因上发生突变，这可能是用通常的方法识别不切实际的。 该项目的目标是开发一种全面的基因表达和互补蛋白图谱方法，利用通过IRP批准的方案(2003-086)从受影响患者那里收集的样本。假说是，在并发症方面具有相似表型的条件，无论潜在的基因突变如何，其表达和蛋白质组谱将被证明是相似的，并适用于使用相同靶点的治疗。例如，Loeys-Dietz综合征是由细胞表面受体TGFbetaR1&2突变引起的，而马凡综合征是由FBN1突变引起的，FBN1是一种结构性细胞外基质蛋白。尽管导致突变的原因不同，但这两种综合征都会导致TGFβ途径的紊乱，在表型上有相似之处，并且可以接受氯沙坦的治疗，氯沙坦是一种血管紧张素受体阻滞剂，也是TGFβ表达的调节剂。 我们正在利用全基因组表达阵列、RT-PCR2-D蛋白质电泳、质谱学、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的患者的情况，如口蹄疫。最近的研究结果表明，马凡综合征、VEDS和FMD患者循环中的TGFbeta1水平升高。我们对马凡综合征的研究结果已经发表，其他疾病正在准备发表。鉴于实验室正在研究300多个来自结缔组织患者的独特的患者成纤维细胞系，这被视为一个长期项目。

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.