Patient-specific targeting of uterine fibroids

项目摘要 子宫平滑肌瘤（纤维瘤）是育龄妇女最常见的肿瘤， 50岁时患病率高达75%。具有临床显著纤维瘤负担的患者可具有多个 使子宫肌瘤成为美国子宫切除术的主要适应症。有一个 这种疾病的种族差异很大，非洲裔美国妇女的发病时间较早， 疾病，具有更大的严重性，并且到50岁时的患病率为89%。疾病的病因学 在很大程度上是未知的，但最近发现MED 12基因在50-70%的肌瘤中突变， 在小鼠子宫中引入类似突变可导致纤维瘤，这表明突变MED 12可能是一个关键 子宫肌瘤的发病机制然而，在MED 12中， 突变型肌瘤或MED 12野生型肌瘤是未知的。这种知识上的差距阻碍了 开发有效的治疗方法，消除妇女进行直肠切除术的需要。我们建议 利用下一代测序技术，对表观遗传、基因组和 不同亚型的子宫肌瘤的转录景观与正常子宫肌层比较。我们将 我还比较了白种人和非洲裔美国妇女的子宫肌瘤，试图了解子宫肌瘤的差异。 这两个群体之间的疾病，目前没有任何高度显着的遗传相关性， 突变我们已经组建了一个组织处理和分析专家团队， 测序和生物信息学分析，并进行了初步研究，使我们 假设有症状的女性需要一种纤维瘤亚型特异性的精确医学方法， 疾病我们将在一个更大、更多样化的女性队列中验证我们的初步结果， 深度和确定是否升高的血清水平的特征蛋白，我们已经确定是高度 在纤维瘤的特定亚群中表达，可用作该疾病的血清生物标志物。我们将执行 体外和体内实验，以了解其破坏的机制和途径， 过度表达我们将使用人类甲基化（850 k/EPIC）DNA甲基化阵列来确定是否 纤维瘤亚型之间的甲基化差异有助于通过RNA-seq的差异表达。 将在MED 12突变体和HMGA 2之间进行染色质免疫沉淀测序（ChIP-Seq） 过表达肌瘤和子宫肌层细胞，以比较和对比这些染色质的差异结合 修饰蛋白质。我们将使用ATAC-seq来确定子宫肌瘤组织中的开放染色质区域， 与RNA-seq相关。我们将分析广泛和深入的数据，以提供更好的 了解在子宫内膜异位症亚群中发现的组合RNA-seq，表观遗传学和基因组景观 纤维瘤我们希望这些研究将导致患者特异性/精确医学的发展 子宫切除术的替代方法来管理这种疾病。

Project Summary Uterine leiomyomas (fibroids) are the most common tumors found in reproductive aged women, with an overall prevalence of up to 75% by age fifty. Patients with a clinically significant fibroid burden can have multiple debilitating symptoms, making fibroids the leading indication for hysterectomy in the United States. There is a strong racial disparity in the disease, with African-American women presenting with an earlier onset of the disease, with greater severity, and having a prevalence of 89% by the age of fifty. The etiology of the disease is largely unknown, but recent discoveries that the MED12 gene is mutated in 50-70% of fibroids and that introduction of a similar mutation in mouse uteri can lead to fibroids suggest that mutant MED12 may be a key player in the etiology of uterine fibroids. However, the actual mechanisms driving the disease, in either MED12 mutant fibroids or MED12 wildtype fibroids, are unknown. This gap in knowledge has hindered the development of effective therapies that obviate the need for women to have hysterectomies. We propose to exploit next generation sequencing to perform comprehensive analyses of the epigenetic, genomic, and transcriptional landscape of different subsets of uterine fibroids for comparison with normal myometria. We will also compare fibroids from caucasian and African-American women to try to understand the disparity in the disease between these two populations, which are not currently associated with any highly significant genetic mutations. We have assembled a team of experts for tissue processing and assays, next generation sequencing, and bioinformatic analyses and have performed preliminary studies that have led us to hypothesize that a fibroid subtype-specific precision medicine approach is needed for women with symptomatic disease. We will validate our preliminary results in a larger and more diverse cohort of women with greater depth and determine whether elevated serum levels of characteristic proteins we have determined are highly expressed in specific subsets of fibroids could be used as serum biomarkers for the disease. We will perform in vitro and in vivo experiments to understand the mechanisms and pathways disrupted by their overexpression. We will use the HumanMethylation (850k/EPIC) DNA methylation array to establish whether methylation differences among the fibroid subtypes contribute to the differential expression by RNA-seq. Chromatin immunoprecipitation sequencing (ChIP-Seq) will be done between MED12 mutant and HMGA2 overexpressing fibroids and myometrial cells to compare and contrast differential binding of these chromatin modifying proteins. We will use ATAC-seq to determine open chromatin regions in the tissues of the fibroid subtypes and correlate with RNA-seq. We will be analyzing both broad and deep data to provide a better understanding of combined RNA-seq, epigenetics, and genomic landscapes found in subsets of uterine fibroids. We expect that these studies will lead to development of patient-specific/precision medicine alternatives to hysterectomy for management of this disease.