2016-01-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/645798摘要： 高血壓與其造成的血管損傷如動脈瘤為公共衛生上的重要威脅。通過一個針對高血壓進行的大型家族世代（SAPPHIRe）全基因組掃描，我們找到了RRBP1上的的遺傳變異（核糖體結合蛋白-1）與的收縮壓（P = 9X 10-6）和舒張壓（P = 1.9×10-4）有強烈相關。這一發現亦獲得後續三個大規模的全基因組關聯研究（GWAS）證實。令人意外的, 獨立的GWAS也發現RRBP1為動脈瘤的致病基因。 我們發現， rrbp1基因剔除鼠小鼠血壓顯著下降（收縮壓：119.5 vs. 135.1毫米汞柱，P <0.01; 舒張壓：91.2毫米汞柱 vs. 109.1mmHg，P <0.01）, 伴隨心肌收縮力改善（心輸出量21.1毫升/分鐘vs. 16.8毫升/分鐘，P = 0.03）, 以及一氧化氮合成酶的表達增加。顯示rrbp1缺乏具有增加心臟收縮力與放鬆血管的效果。 在注射血管張力素II的小鼠身上, rrbp1在主動脈的表現量也與主動脈擴張的的程度成正相關。這些數據表明，抑制rrbp1是治療高血壓與主動脈瘤的一個重要的治療目標。我們發現Rrbp1主要表現在血管內皮細胞。 rrbp1 在粗內質網會調節一組特定的基因的mRNA的表現。轉錄體分析發現，Rrrp1的缺乏顯著增加gde7的表達（glycerophosphodiesterase-7，也稱為gdpd3）, gde7為一種新型的gde家族成員, 具有磷脂酶D活性, 會裂解細胞膜上的lysophosphatidylcholine (LPC) 與 lysophosphatidic acid (LPA)。 LPA已被證明通過結合LPA受體和激活Rho磷酸酶 /肌球蛋白II, 而調節血管內皮細胞的功能，移動與修復。在這裡，我們計劃在3年內：1.闡明rrbp1剔除降低小鼠血壓與減少主動脈瘤生成的分子機轉。2.探討內皮組織特異性與全身誘導性rrbp1基因剔除小鼠的表現型。3.證明rrbp1脂質體siRNA治療高血壓與修復主動脈瘤的療效。4.測試使用gde7腺相關病毒載體降低血壓與修復主動脈瘤的效果。5. 測試血漿中GDE7濃度與高血壓與動脈硬化的相關; 以及其預測動脈硬化進展的能力。6.以高通量小分子篩選GDE7酶調節劑, 以臨床前試驗其對高血壓與主動脈瘤的治療效果。<br> Abstract: Hypertension and associated vascular injury such as arterial aneurysm are major threat to public health. Through genome-wide scan for hypertension in a large family cohort (the SAPPHIRe study), we identified genetic variants of RRBP1 (ribosome binding protein -1) are strongly associated with systolic blood pressures (SBP, P=9x 10-6) and diastolic blood pressures (DBP, P=1.9 x 10-4). This finding was confirmed by three large genome-wide association studies (GWAS) for blood pressures. Unexpectedly, independent GWAS also revealed RRBP1 as a susceptible gene for arterial aneurysm. We found that rrbp1-deficient mice had significantly lowered blood pressure (SBP: 119.5 vs. 135.1 mmHg, P< 0.01; DBP: 91.2 mmHg vs. 109.1mmHg, P< 0.01), improved myocardial contractility (cardiac output: 21.1 ml/min vs. 16.8 ml/min, P=0.03), as well as increased nitric oxide synthase expressions compared to wild-type mice, indicating an inodilatory effect. The expression of rrbp1 in aorta also correlates with the extent of aortic dilation in mice infused with angiotensin-II. These data indicate inhibition of RRBP1 is an important therapeutic target for hypertension and aortic aneurysm. Rrbp1 is expressed in vascular endothelium with subcellular localization in rough endoplasmic reticulum. Rrbp1 have been shown to regulate the mRNA metabolism of a specific set of genes. Transcriptome analysis showed that deficiency of rrbp-1 markedly increase the expression of gde7 (glycerophosphodiester phosphodiesterase 7, also named gdpd3), a novel member of the gde family with phospholipase D activity cleaving membrane phospholipid lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA). LPA have been shown to modulate endothelial cell function, migration and tissue repair via binding to LPA receptor and activation of Rho receptor/myosin II. Here we proposed in the following 3 years to:1. Investigate the molecular mechanism by which rrbp1-knockout mice are protective of hypertension and aortic aneurysm.2. Dissect the phenotype of endothelium-specific and inducible rrbp1 knockout mice3. Prove the therapeutic potential of rrbp1 liposome- siRNA injection on hypertension and repairing of aortic aneurysm in mice.4. Test the therapeutic effects using adeno- associated viral (AAV) vectors expressing gde7 for repairing aortic aneurysm and reducing blood pressure.5. Examine the association of levels of GDE7 in serum with hypertension, atherosclerotic disease and explore their prognostic/diagnostic value.6. High-throughput small-molecule screening to identify enzymatic modifier for GDE7 and preclinical test for their therapeutic effect on aortic aneurysm and hypertension高血壓主動脈瘤遺傳hypertensionaortic aneurysmgenetics