2017-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/710943摘要：化療 (chemotherapy, CTx) 經常被使用於治療各種惡性腫瘤，然而，化療會導致女性病患發生永久性的卵巢早衰 (premature ovarian failure, POF) （平均 50% 之發生率）。吾人先前已證實羊水幹細胞 (amniotic fluid stem cells, AFSCs) 移植能夠於化療藥物誘導卵巢早衰的小鼠疾病模式中修復受損卵巢的功能，唯其修復機制仍未明。此外，臨床上對於使用幹細胞移植療法仍存有高度的安全性以及道德爭議。據此，尋找新的治療方式以防止卵巢早衰是目前臨床上之重要議題。研究發現，幹細胞可透過其分泌之外切體 (exosomes) 修復受損組織。本研究室先前之體外試驗研究也證實，羊水幹細胞分泌之外切體能夠傳遞 RNAs 至受損的卵巢顆粒性細胞 (granulosa cells, GCs) 以達到防止受損細胞凋亡之效果。有鑑於此，釐清幹細胞分泌之外切體和目標細胞間之交互作用將有助於發展新的策略於臨床上治療卵巢早衰。 為此，本研究室擬先進行體內試驗以確認羊水幹細胞之外切體於化療藥物誘導卵巢早衰的小鼠疾病模式中可藉由傳遞 RNAs 之方式修復受損卵巢的功能。其後，本研究室計畫使用次世代定序 (next generation sequencing, NGS) 分析羊水幹細胞之外切體內 RNA 之表現型 (expression profile)，並以此資料庫篩選出可能參與治療機制之功能性 RNAs。為研究此等功能性 RNAs 之功能，吾人將設法降低羊水幹細胞之外切體中候選 RNAs 之表現，並以體外試驗檢驗此羊水幹細胞之外切體之治療效果是否受到影響。吾人下一步擬使用人工合成之微脂粒 (liposomes) 將候選 RNAs 直接送入受損的卵巢顆粒性細胞內以驗證其療效。最後，吾人計劃直接使用微脂粒將此等候選 RNAs 施行於化療藥物誘導卵巢早衰的小鼠疾病模式以評估其治療效果。本計畫之研究成果將會闡明羊水幹細胞之治療機轉，並暗示在臨床上藉由新穎的非細胞 (cell-free) 療法，達成有效治療卵巢早衰之可行性。<br> Abstract: Chemotherapy (CTx) is a common way to fight against various cancers but causes a high incidence (average 50%) of irreversible premature ovarian failure (POF) in female patients by the end of treatment. Although we have shown that amniotic fluid stem cells (AFSCs) can preserve most ovarian function in CTx-induced POF mouse model, the restoring mechanism of AFSCs transplantation is elusive. In addition, clinical use of stem cell transplantation in human is highly controversial about the safety and ethical issues. In view of this, it is necessary to find an alternative treatment for the prevention of POF in clinic. Exosomes released from stem cells have been reported as a novel mechanism involved in the beneficial effects of stem cell therapy. In our previous in vitro study, we further proved that the RNAs delivered by AFSC-derived exosomes play a major role in the effect of AFSCs that suppress the apoptosis of CTx-damaged ovarian granulosa cells (GCs), which are required for oocyte survival and follicle development. Accordingly, the recognition of exosomes-mediated interaction between stem cells and target cells may found new strategies for treating POF in clinic. To address this issue, we will first validate whether AFSC-derived exosomes contribute to the preservation of ovarian follicles in CTx-induced POF mouse model via delivery of RNAs. Next, we plan to perform next generation sequencing (NGS) to analyze the RNAs expression profiles of AFSC-derived exosomes so that we expect to identify the candidate RNAs involved in the AFSCs-based restoration. To test the function of the candidate RNAs, we plan to knock down the expression of candidate RNAs in AFSC-derived exosomes to test whether the effects of AFSC-derived exosomes will be perturbed in our in vitro study. Further, we will directly deliver the candidate RNAs via artificial liposomes into CTx-damaged GCs to verify their therapeutic effects. Finally, we aim to administrate the liposomes with candidate RNAs directly into CTx-induced POF mouse model and assess their effectiveness. These studies will uncover the restorative mechanism of AFSC-based therapeutics and imply the possibility of a novel cell-free therapeutics for treating POF.卵巢早衰再生醫學羊水幹細胞外切體Premature ovarian failureRegenerative medicineAmniotic fluid stem cellsExosomes