2016-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/648268摘要：多囊性卵巢症候群(PCOS)為生育年齡婦女最常見的內分泌障礙，表現之症狀為月經不規則、不排卵、雄性素過高、多囊性卵巢，是造成不孕症的重要原因之一。PCOS 之病人常併發代謝症候群，然其致病機轉至今仍未明。近來研究顯示顆粒細胞與濾泡膜細胞之功能失調造成濾泡之不正常形成以及雄性素過高；儘管顆粒細胞在多囊性卵巢症候群的致病機轉扮演著關鍵的角色，目前為止針對人類顆粒細胞的研究卻十分有限，主要原因之一是取得極為困難，過去的研究只能利用卵巢手術時取得檢體再進行體外細胞培養，體外培養存活期亦不長因而使得研究更加困難。誘導型多功能幹細胞 (iPSC) 使研究者可以在培養皿中模擬複雜性人類疾病。我們的團隊已成功製造出人類多囊性卵巢症候群之專一性iPSCs，更進一步發展出可將ESCSs/iPSCs 分化成類顆粒細胞的方法，使PCOS 之體外表現型之分析更為容易，利用此模式來研究PCOS 之致病機轉。本研究計畫的目的是利用誘導多囊性卵巢症候群病患之誘導多潛能幹細胞(iPSC)分化成為具有功能性之卵巢顆粒細胞，建立一個有效的模型來探討卵巢顆粒細胞分化生長、濾泡及卵子之發育成熟、類固醇之製造調控與多囊性卵巢症候群之致病機轉等。更進一步，將不同臨床表現的多囊性卵巢症候群病患之誘導型多功能幹細胞分化為卵巢顆粒細胞，分析病患卵巢顆粒細胞與正常人之基因表現，期能瞭解此疾病，並發展出更佳之診斷模式及具病患專一性之治療方法及藥物篩檢平台。我們將正常人與 PCOS 病人的iPSC 在分化成類顆粒細胞後，以qRT-PCR 分析六個顆粒細胞標示基因 (AMH、FOXL2、FSHR、AMHR2、LHR 及CYP19A1) 在不同的類顆粒細胞中表現之差異，FOXL2 的表現量在PCOS 病人iPSC 分化的類顆粒細胞顯著的比正常人iPSC 分化的類顆粒細胞低，研究指出FOXL2 會影響卵巢的發育與功能，同時也跟顆粒細胞造成的病理相關。本研究計畫預計收集接受試管嬰兒技術治療的多囊性卵巢症候群病患周邊血液及濾泡液中的顆粒細胞，並進一步區分四種不同PCOS 的表現型，分析FOXL2 表現量的差異，預期能進一步了解FOXL2 基因表現在PCOS 致病機轉所扮演的角色。<br> Abstract: Polycystic ovarian syndrome (PCOS) is one of the most common endocrine disorders inwomen at reproductive age. PCOS is characterized by chronic anovulation, menstrual irregularities,polycystic ovaries, and hyperandrogenism. However its pathophysiology is still unclearand.Therefore the progress in diagnosis and treatment is limited. In previous research, one of the mostcritical pathophysiology of PCOS was considered to be hyperandrogenism. Recent studies havesuggested that dysfunction of granulosa cells and theca cells contribute to the abnormalfolliculogenesis and hyperandrogenism in PCOS.Although the granulosa cells might have important role in PCOS and other hyperandrogenemicdisorders, our understanding toward granulosa cells is very limited. Granulosa cells that can beobtained in human are most from transvaginal oocyte retrieve. With the advent of the technique ofinduced pluripotent stem cells (iPSCs), we have the opportunity to model complex human diseasesin culture dishes. Our group has successfully derived disease-specific iPSCs from human PCOS.Furthermore, our group has successfully developed a method to efficiently differentiate humanESCs/iPSCs into granulosa-like cells, making the in vitro PCOS studies more feasible.In this proposal, we will differentiate the induced pluripotent stem cells from women of variousphenotypes’ PCOS into granulosa cells. Through these we will have better understand the biogenesisof ovarian steroids hormone, the growth and maturation of follicles, the gene expression ofgranulosa cells, and even the pathogenesis of PCOS. Furthermore, the discovery of new drugs ortreatment for PCOS will be tested in this disease specific PCOS model.In the previous study, we have analyzed GC marker gene’s expression in the differentiatedgranulosa-like cells from both non-PCOS amd PCOS iPSCs. Among 6 GC maker genes, AMH、FOXL2、FSHR、AMHR2、LHR and CYP19A1, only FOXL2 showed significant low expression indifferentiated granulosa-like cells from PCOS iPSCs. FOXL2 has been reported involving in ovaryfunction and maintenance, and granulosa cell proliferation. We plan to investigate the FOXL2expression in different phenotypes’ PCOS, and get more insights of FOXL2 in the mechanisms ofPCOS.多囊性卵巢症候群顆粒細胞誘導型多能幹細胞polycystic ovarian syndromegranulosa cellsinduced pluripotent stem cells