靜脈曲張為常見靜脈疾病。在正常靜脈，內皮細胞與胞外間質及平滑肌細胞作用，並與平順血流接觸維持恆定。血液逆流與胞外間質重組發生於靜脈曲張促進靜脈病變，但兩者間互動對靜脈病變的影響仍屬未知。近年來，非編碼核糖核酸被發現為血流調控因子影響動脈病變，但在靜脈曲張尚未研究。我們初步結果顯示非編碼核糖核酸及發炎訊息可能與血液逆流誘導靜脈內皮或平滑肌病變相關，因此本研究發展共培養流體系統釐清正常靜脈平順流vs.血液逆流擾流、胞外間質重組、及內皮/平滑肌細胞溝通對靜脈非編碼核糖核酸訊息影響，將使用次世代定序、生物資訊、大量表現/剃除技術全面尋找嶄新血液逆流調控的靜脈內皮或平滑肌非編碼核糖核酸及其功能；靜脈內皮或平滑肌細胞貼附的胞外間質與細胞間溝通對這些血液逆流調控非編碼核糖核酸訊息的影響亦將被研究。不同逆流程度及不同病變級別的人類病變血管將被收集驗證體外實驗結果，且血液中非編碼核糖核酸將被評估發展為診斷分子。PEG化奈米酯小體將被發展輸送非編碼核糖核酸抑制血液逆流誘導靜脈病變。本計畫將提供微環境、血液逆流及細胞間溝通調控非編碼核糖核酸靜脈病變訊息的新知，可發展診斷治療方式及組織工程，具有創新性及臨床應用潛力。Varicose vein or chronic venous disease (CVD) is the common venous disorder with high prevalence and recurrence rate. The cost of treatment and healthcare for varicose vein makes the huge financial burden in Taiwan. In normal vein, venous endothelial cells (ECs) react to extracellular matrix (ECM), communicate with venous smooth muscle cells (SMCs), and are exposed to blood laminar flow to maintain venous homeostasis. However, blood reflux and ECM remodeling are happened on varicose vein to promote venous disorders. Although blood reflux and ECM remodeling both have been identified to be the vital pathogenic factors for varicose vein, there is no study to identify the role of their interaction in regulating venous dysfunctions. Recently, non-coding RNAs (ncRNAs), i.e., long ncRNAs and miRNAs, have emerged as novel shear-sensitive molecules to affect aortic functions, but the roles of ncRNAs in varicose vein remain unknow. Our preliminary results demonstrated that shear-sensitive ncRNA signaling, e.g., miR-10a signaling, and inflammatory signaling, e.g., Smad signaling, can be modulated by blood reflux in both venous ECs and SMCs of human varicose vein. This supports that blood reflux may regulate ncRNA signaling to affect venous EC or SMC dysfunctions in varicose vein. In this proposed research, we will develop in vitro co-culture flow system, which can co-culture venous ECs and SMCs on different ECM and expose EC side to normal laminar flow vs. blood reflux-indced oscillatory flow, to elucidate the relationship between two vital pathogenic factors, i.e., blood reflux and ECM remodeling, and EC/SMC communication in modulating ncRNA signaling and venous cell dysfunctions in varicose vein. The state-of-the-art approaches, including high throughput next generation sequencing, bioinformatics, and ncRNA overexpression/knockdown assay will comprehensively identify novel blood reflux-modulated venous EC or SMC ncRNAs and their involved signaling pathway and venous dysfunctions. Moreover, the roles of ECM composition or remodeling around venous EC or SMC and EC/SMC communication in blood reflux-modulated ncRNA signaling and venous cell dysfunctions also will be elucidated. In addition, human specimens, including human varicose veins with different level of blood reflux or human CVD veins with different CEAP stage vs. human normal veins, will be collected to confirm our in vitro results in vivo. The levels of identified blood reflux-modulated ncRNAs in serum will be detected to identify their potential for varicose vein diagnosis. Finally, PEGylated cationic nanoliposome will be developed to deliver precursor or antagonist of blood reflux-modulated ncRNA to inhibit blood reflux-induced venous EC or SMC dysfunctions. The results obtained in this study provide new knowledge on the relationship between microenvironment, blood reflux, and venous EC/SMC communication in modulating ncRNA signaling to affect venous dysfunctions in varicose vein. PEGylated cationic nanoliposome carrying ncRNA and serum ncRNAs will be developed as novel therapeutic and diagnostic strategies for venous disorders, respectively. In addition, this study will identify which ECM can inhibit blood reflux-induced venous cell dysfunctions. The information will help us to develop tissue engioneering for varicose vein treatment. Thus, the findings of this study are of high novelty with great potential for clinical applications.
extracellular matrix remodeling
venous endothelial cell/smooth muscle cell communication