2016-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/646213摘要：近年來基礎與臨床的研究都顯示，急性腎損傷後腎臟可能修復不完全而導致慢性腎臟病，因此有必要詳細探討急性腎損傷轉換為慢性腎臟病（AKI-to-CKD transition）的病理機轉，找出可改變的因子以發展治療策略。富含半胱氨酸61蛋白（Cysteine-rich protein 61,Cyr61）會在正常與疾病的腎臟中有不同程度的表現。吾人過去的研究顯示Cyr61在急性與慢性腎臟疾病中扮演關鍵角色，聯繫了腎小管上皮細胞的傷害、過度的修復作用、發炎反應、與微血管循環稀薄現象，進而導致進行性的纖維化。我們初步實驗發現Cyr61的表現在不同的急性腎損傷動物模式中並不相同，而較嚴重慢性腎臟病患者尿中Cyr61的排出量比早期慢性腎臟病患者顯著增加。吾人因而推論急性腎損傷後腎臟Cyr61參與了發炎反應並導致慢性腎臟纖維化，並據此設計了三年期研究計畫。第一年期將研究Cyr61在三種分別導致不同纖維化程度的急性腎損傷動物模式中的表現差異，用以發展其可能的臨床用途。其次，將以具有Cyr61與connective tissue growthfactor雙重目標阻斷功能的抗體治療阻塞性腎臟纖維化模式，了解這兩種重要因子在腎臟纖維化的交互作用。第二年期，吾人計畫培育可誘導性腎小管上皮細胞專一性的Cyr61基因剔除轉殖鼠（Pax8-rtTA/LC-1, Cyr61Flox/Flox），用以研究急性腎損傷後的恢復期，Cyr61對腎小管細胞的傷害與再生、組織發炎反應、間質纖維化、微血管循環稀薄現象、與巨噬細胞極化方向的影響。第三年期，將使用前述的基因轉殖鼠，研究Cyr61對於急性腎損傷後造成進行性慢性腎臟病之長期生理與組織病理的影響。並利用lentivirus轉染製造Cyr61過度表現的腎小管上皮細胞株，研究Cyr61的訊息傳遞途徑。吾人期望藉由此計畫的各項實驗結果，能進一步瞭解AKI-to-CKD transition的機轉，並據此發展新的診斷與治療策略。<br> Abstract: Clinical and basic studies have suggested that acute kidney injury (AKI) would result inincomplete repair and lead to chronic kidney disease (CKD). Understanding the pathophysiology ofAKI-to-CKD transition and identifying potential modifiable factors are warranting. Cysteine-richprotein 61 (Cyr61), a matrix-associated protein, expresses in normal and diseased kidneys. Ourprevious studies demonstrated that the major role Cyr61 plays in acute and chronic kidney disease islinking tubular epithelium injury with inflammation, maladaptive repair, capillary rarefaction, andprogressive kidney fibrosis. Our preliminary experiments for this project showed that the renal Cyr61expression pattern is different in various AKI model. The urinary Cry61 concentrations in patientswith advanced CKD are significantly higher than those in patients with earlier stages of CKD.Therefore, we hypothesized that renal Cyr61 upregulation after AKI participates to the inflammatoryprocess and leads to chronic kidney fibrosis. Thus we design this three-year project to investigate thepathogenetic mechanism of Cyr61 in AKI-to-CKD transition.In the first years, we will evaluate the expression profiles of Cyr61 in various AKI models withdifferent fibrotic outcomes. This may help to develop clinical usage of Cry61 to predict AKI patients’outcome. Subsequently, we will block both Cyr61 and connective tissue growth factor in the unilateralureteral obstruction model to clarify the interaction of these two important pathogenetic factors duringrenal fibrosis after AKI. In the second year, we plan to generate doxycycline-dependent tubularepithelial cell-specific Cyr61 gene knock out mice (Pax8-rtTA/LC-1, Cyr61Flox/Flox mice). To study theeffect of Cyr61 on kidney recovery after an acute injury, we will investigate the changes in tubularinjury, tubular regeneration, tissue inflammation, interstitial fibrosis, and capillary rarefaction by usingthis genetically modified mice in the model of AKI with recovery. The effect of Cyr61 on macrophagephenotype changes will also be investigated. In the third year, we will define the long-termpathological and physiological effect of Cyr61 on kidneys after an AKI we could investigate theeffects of Cyr61 on the AKI-to-CKD transition in the genetically modified mice after an AKI withprogressive consequences. Finally, we plan to produce Cyr61-overexpressed cell by lentivirustransfection and investigate the signal transduction pathway of Cyr61 on renal tubular epithelial cellsin vitro. We believe that this project could discover new insights in the post-AKI pathogenesis.Hopefully, it will expand the knowledge of the mechanism of progressive kidney fibrosis andAKI-to-CKD transition. The results could contribute to development of novel diagnostic andtherapeutic strategies against the gradual kidney function decline.