摘要:繆氏腺肉瘤是罕見的女性生殖道惡性腫瘤,其組成為混合上皮和間質成分構成。其上皮成分是良性或是非典型的,但其基質成分的表現為低惡性。如果腫瘤體積有四分之一以上的由高惡性肉瘤細胞組成,腫瘤會稱為"具肉瘤過度增生之腺肉瘤",其臨床惡性度較高。在臺灣,它占所有原發性子宮肉瘤的5%左右。根據全國性調查的結果,好發年齡平均是 46 歲左右,有大約一半的病人在生殖年齡。然而,全球資料顯示這種腫瘤主要發生在停經後的婦女,雖然發生在生殖年齡病人約占 30%。外觀上,大多數病灶通常是息肉狀,平均直徑 6.5 釐米。 組織病理學上,息肉狀突起由高細胞密度之基質細胞與拉長,壓縮的腺體所組成,並伸入擴張的腺體腔中。子宮切除術為治療子宮腺肉瘤主要方法。雖然整體預後不差 (5 年與10 年存活率大約分別為77%和 58%),它對女性生殖健康和婦女的生育能力有很大影響。為了使保留生育能力的治療成為可能,發展出選擇性的、 有針對性的治療方法的無疑是我們的主要目標之一。繆氏腺肉瘤是很難收集新鮮腫瘤標本進行研究的,因為其組織學的變化是很細微的,而且廣泛的取樣是作出正確診斷所必要。最近,我們利用 Oncoscan FFPE 檢測,剛剛完成了繆氏腺肉瘤全基因拷貝數變化分析。在全基因拷貝數變化中,CDK4 和MDM2基因的拷貝數增加頻繁地發生。外國的研究人員報導了類似的結果。我們還發現頻繁大量染色体的破碎和重组(chromothripsis) 在肉瘤過度增生之腺肉瘤常發生。若缺乏對應的基因表現研究,這些結果的意義是有限的。此外,現今尚未有繆氏腺肉瘤的基因表現研究,缺乏這種腫瘤類型全面概述性分子方面的研究,更使發展標靶治療成為空談。本研究的內容是要將兩個新開發的技術應用:Nanostring nCounter 基因表現檢測和Duolink in situ PLA (Proximity Ligation Assay) 檢測來應用在 測定繆氏腺肉瘤福馬林固定標本上。我們希望透過這些兩個化驗,得到信使RNA 和蛋白質定量或半定量的表現資料。具體目標一: 利用Nanostring nCounter 基因表現分析,我們計畫得到信使RNA定量或半定量的表現資料。我們將比較這一結果與我們以前全基因組拷貝數變化分析的資料,找出一致的生化變化。具體目標二: 使用Duolink in situ PLA 檢測,我們打算由蛋白表現層面,驗證從第一個具體目標中最重要的發現。我們從這些研究中獲得的結果,將揭示腫瘤的發生與惡化相關的新分子病因,並很有可能會提供一套針對繆氏腺肉瘤針對預後的預測和進行藥物治療的新標靶的遺傳標記。
Abstract: Mullerian adenosarcoma is an uncommon gynecologic malignancy composed of mixedepithelial and mesenchymal components. Their epithelial components are benign or atypical,but their stromal components behave as low malignant elements. If one fourth of the tumorvolume is composed of high-grade sarcomatous cells, the tumor will be classified as“adenosarcoma with sarcomatous overgrowth”, which tend to behave more aggressively.In Taiwan, it accounts for roughly 5% of all primary uterine sarcomas. According to anation-wide, population-based surveillance, the average age of involvement is around 46years, with about half of the patients within their reproductive ages. Nevertheless, theworldwide data show that this tumor occurs primarily in postmenopausal women, althoughabout 30% are found in their reproductive ages. Macroscopically, most lesions are typicallypolypoid with a mean diameter of 6.5 cm. Histopathologically, polypoid projectionscomposed of hypercellular stroma and elongated, compressed glands protrude into the dilatedglands. Uterine adenosarcoma is treated by hysterectomy. Although the overall prognosis isbarely satisfactory (with 5-year and 10-year survival rates of approximately 77% and 58%,respectively), it has great impact on female reproductive health and a woman’s ability to bearchildren. In order to make fertility-sparing therapeutics possible, the development of selective,targeted therapies is undoubtedly one of our main goals.It is very difficult to harvest fresh tumor samples of Mullerian adenosarcomas forresearch because their histologic changes are very subtle, and extensive sampling is necessaryfor making the correct diagnosis. Recently, we just finished genome-wide profiling of copynumber variations of Müllerian adenosarcomas using Oncoscan FFPE assay. Copy numbergains of CDK4 and MDM2 genes are noted between frequently occurring copy numberchanges. Similar results are reported by foreign researchers. We also found frequentchromothripsis in those cases with sarcomatous overgrowth. The significance of thesefindings is limited due to the lack of corresponding expression studies. Furthermore, therehave been no recorded expression studies in the general literature, hence the lack of acomprehensive overview of the molecular aspects of this tumor type, let alone thedevelopment of targeted therapy.The objective of this study is to apply two newly-developed technologies: NanostringnCounter gene expression assay and duolink in situ PLA (Proximity Ligation Assay) assay onFFPE specimens of Müllerian adenosarcomas. Through these two assays, we hope to havequantitative or semi-quantitative expression data of mRNAs and proteins.Specific aim1: Using the technique of Nanostring nCounter gene expression assay, weplan to harvest quantitative or semi-quantitative expression data of mRNAs. We will comparethis result with our previous genome-wide profiling of copy number variations to find out thecoordinating biochemical changes.Specific aim# 2: Using the technique of duolink in situ PLA assay, we plan to validatethe most significant findings from the first specific aim in protein expression level.Our results obtained from these projects will shed a new light on the molecular etiologyof tumorigenesis and progression, and will quite likely provide a new set of genetic markersfor outcome prediction and novel targets for drug intervention to treat Müllerianadenosarcomas.