Elucidating a novel WNT4 regulatory axis as a driver of gynecologic cancer health disparities

项目摘要/摘要 卵巢癌等妇科恶性肿瘤是影响女性的最致命的癌症之一。 由于治疗阻力和对疾病病因和风险的有限了解。未被充分发掘的风险因素 是Wnt配体Wnt4，它是卵巢器官发生的中心。20多项研究表明Wnt4基因多态 增加患妇科疾病的风险；一个关键的Wnt4调节位点的多态性(Rs3820282) 与卵巢癌风险增加10-25%相关，但其机制(S)尚不清楚。我们的工作将WNT4链接到 癌细胞的生长、新陈代谢和治疗耐药性。我们发现WNT4的过度表达足以调节 体外化疗耐药，体内转移生长增加的耐药，以及WNT4 在新辅助化疗后存活的OvCa细胞中有强烈的诱导表达。重要的是，rs3820282 WNT4调节位点上的变异等位基因为核受体类转录因子创建了一个结合位点。 在小鼠中，rs3820282变异的CRISPR敲入增加了WNT4在妇科组织中的表达。 因此，在对100多个OvCa肿瘤组织的蛋白质阵列研究中，我们发现AMPK激活和 下游信号转导在变异等位基因肿瘤中增加。相反，葡萄糖代谢蛋白是 野生型肿瘤中升高，与AMPK信号转导负相关，提示为Wnt4基因 支持新陈代谢重塑。这些观察结果表明，rs3820282变异体激活WNT4以 导致癌症表型。然而，rs3820282变异等位基因频率(Vaf)在 种族人口，在非洲人口中约为0%，在高加索人中约为15%，在拉丁裔人口中为20%-40% 亚洲人群中45%-55%，与侵袭性和治疗耐药性的高发病率平行 亚洲人群OvCa亚型透明细胞癌(CCC)我们的目标是确定rs3820282 调节卵巢癌预后的差异，机械地定义由基因型驱动的肿瘤病因，以及 确定开发对WNT4依赖的治疗方法。为了实现这一目标，我们将：1)定义rs3820282 Variant激活依赖Wnt4的代谢重塑；2)定义rs3820282驱动的肿瘤发生和 卵巢透明细胞癌(CCC)模型的治疗反应；3)决定rs3820282基因型 对卵巢癌患者的预后有影响。以来自人类的严格的支持数据为基础 标本，我们将进行高度机械化的研究，以确定这一共同的贡献 多态与癌症差异、肿瘤代谢重编程、妇科肿瘤发生、治疗 反应，以及患者的结果。我们将利用尖端的全球新陈代谢组学、肿瘤发生模型、 和人类生存研究。我们的方法可以定义基因-表型之间的联系，确定这个SNP是如何 推动卵巢癌的差异，并确定利用潜在生物学的方法。

PROJECT SUMMARY/ABSTRACT Gynecologic malignancies such as ovarian cancer (OvCa) are among the deadliest cancers affecting women due to therapy resistance and limited understanding of disease etiology and risk. An under-explored risk factor is Wnt ligand WNT4, which is central to ovarian organogenesis. Over 20 studies link WNT4 polymorphisms with increased risk for gynecologic pathologies; one polymorphism at a key WNT4 regulatory site (rs3820282) is associated with 10-25% increased risk for OvCa, but the mechanism(s) is unknown. Our work links WNT4 to cancer cell growth, metabolism, and therapy resistance. We find WNT4 over-expression is sufficient to mediate chemotherapy resistance in vitro, and resistance with increased metastatic outgrowth in vivo, and that WNT4 expression is strongly induced in OvCa cells surviving neoadjuvant chemotherapy. Importantly, the rs3820282 variant allele in the WNT4 regulatory site creates a binding site for nuclear receptor-class transcription factors. CRISPR knock-in of the rs3820282 variant in mice increases Wnt4 expression in gynecologic tissues. Accordingly, in a protein array study of more than 100 OvCa tumor tissues, we found that AMPK activation and downstream signaling were increased in variant allele tumors. Conversely, glucose metabolism proteins were increased in wild-type tumors and inversely correlated with AMPK signaling, suggesting WNT4 genotype underpins metabolic remodeling. These observations suggest that the rs3820282 variant activates WNT4 to drive cancer phenotypes. However, the rs3820282 variant allele frequency (VAF) is widely divergent across ethnic populations, occurring at ~0% in African populations, ~15% in Caucasians, 20-40% in Latinx populations, and 45-55% in Asian populations, paralleling high incidence of aggressive, treatment-resistance OvCa subtype clear cell carcinoma (CCC) in Asian populations. Our goal is to determine how rs3820282 mediates disparities in ovarian cancer outcomes, mechanistically define genotype-driven tumor etiology, and identify therapies to exploit dependence on WNT4. Toward this goal, we will: 1) define how the rs3820282 variant activates WNT4-dependent metabolic remodeling; 2) define rs3820282-driven tumorigenesis and therapeutic response in a model of ovarian clear cell carcinoma (CCC); 3) determine how rs3820282 genotype impacts outcomes for patients with OvCa. With a foundation of rigorous supporting data from human specimens, we will undertake highly mechanistic studies to define the contribution of this common polymorphism to a cancer disparity, tumor metabolic reprogramming, gynecologic tumorigenesis, treatment response, and patient outcomes. We will leverage cutting-edge global metabolomics, tumorigenesis modeling, and human survival studies. Our approach can define the genotype-to-phenotype link, determine how this SNP drives OvCa cancer disparities, and identify approaches to exploit the underlying biology.