Patient-specific targeting of uterine fibroids

项目摘要 子宫肌瘤(肌瘤)是育龄妇女最常见的肿瘤， 到50岁时，患病率高达75%。临床上有显著肌瘤负担的患者可以有多个 使人虚弱的症状，使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，非裔美国女性表现出较早的发病 疾病，病情更严重，到50岁时患病率为89%。这种疾病的病因学 在很大程度上是未知的，但最近的发现MED12基因在50%-70%的肌瘤中发生了突变，而且 在小鼠子宫中引入类似的突变可导致子宫肌瘤提示突变MED12可能是关键 子宫肌瘤病因学方面的专家。然而，在MED12中，驱动这种疾病的实际机制 突变型肌瘤或MED12野生型肌瘤，未知。这种知识上的差距阻碍了 开发有效的治疗方法，使妇女不需要进行子宫切除术。我们建议 利用下一代测序对表观遗传学、基因组和 子宫肌瘤不同亚型的转录图谱与正常子宫肌层的比较。我们会 还要比较高加索女性和非裔美国女性的子宫肌瘤，以试图理解 这两个群体之间的疾病，目前与任何高度显著的基因 突变。我们已经组建了一支组织处理和检测的专家团队，下一代 测序和生物信息学分析，并进行了初步研究，使我们 假设对有症状的妇女需要一种针对肌瘤亚型的精确药物治疗方法 疾病。我们将在更大、更多样化的女性队列中验证我们的初步结果 深度和确定我们已经确定的升高的血清特征蛋白水平是否高度 在特定肌瘤亚群中的表达可作为该病的血清生物标志物。我们将表演 体外和体内实验以了解其干扰的机制和途径 过度表达。我们将使用人甲基化(850k/EPIC)DNA甲基化阵列来确定 不同肌瘤亚型之间的甲基化差异是导致RNA-seq差异表达的原因。 染色质免疫沉淀测序(ChIP-Seq)将在MED12突变体和HMGA2之间进行 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来确定肌瘤组织中的开放染色质区域。 亚型，并与RNA序列相关。我们将分析广泛和深入的数据，以提供更好的 了解在子宫亚群中发现的联合RNA-SEQ、表观遗传学和基因组景观 肌瘤。我们期望这些研究将导致针对患者/精确医学的发展。 子宫切除术的替代治疗这种疾病的方法。

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.