宫颈透明细胞癌(CCC)(cCCC)是一种罕见且侵袭性的人乳头瘤病毒(HPV)阴性宫颈癌,对复发性或转移性疾病的有效治疗选择有限。历史上,下生殖道的CC与子宫内己烯雌酚暴露有关;然而,零星cCCCs的遗传景观仍然未知。在这里,我们试图定义cCCCs的分子基础。使用整个外显子组的组合,有针对性的捕获,和RNA测序,我们在50%(10/20)的CCCs中发现了Hippo信号通路的致病性遗传改变,包括在40%(4/10)的具有Hippo通路突变的病例中反复发生的WWTR1S89W体细胞突变。无论是否存在Hippo途径遗传改变,然而,本研究中分析的所有原发性CCCs(n=20)在转录组和蛋白质水平上都具有Hippo通路失调的特征.体外功能分析显示,WWTR1S89W突变的表达导致TAZ与14-3-3的结合减少,促进TAZ的组成性核易位和Hippo途径抑制。发现WWTR1S89W表达导致获得致癌行为,包括增加扩散,迁移,和体外集落形成以及体内肿瘤发生增加,这可以通过对维替泊芬的TAZ/YAP1复合物的靶向抑制来逆转。最后,表达WWTR1S89W的异种移植物显示出肿瘤表型的转变,变得更具渗透性,分化程度更低,与对照组相比,发现由明显的细胞质清除的细胞组成。我们的结果表明,Hippo途径改变可能是cCCCs的驱动因素,并可能有助于透明细胞表型。针对这种途径的疗法可能构成了一种新的治疗方法,侵袭性肿瘤.©2022英国和爱尔兰病理学会。
Clear cell carcinoma (CCC) of the cervix (cCCC) is a rare and aggressive type of human papillomavirus (HPV)-negative cervical cancer with limited effective treatment options for recurrent or metastatic disease. Historically, CCCs of the lower genital tract were associated with in utero diethylstilbestrol exposure; however, the genetic landscape of sporadic cCCCs remains unknown. Here we sought to define the molecular underpinning of cCCCs. Using a combination of whole-exome, targeted capture, and RNA-sequencing, we identified pathogenic genetic alterations in the Hippo signaling pathway in 50% (10/20) of cCCCs, including recurrent
WWTR1 S89W somatic mutations in 40% (4/10) of the cases harboring mutations in the Hippo pathway. Irrespective of the presence or absence of Hippo pathway genetic alterations, however, all primary cCCCs analyzed in this study (n = 20) harbored features of Hippo pathway deregulation at the transcriptomic and protein levels. In vitro functional analysis revealed that expression of the
WWTR1 S89W mutation leads to reduced binding of TAZ to 14-3-3, promoting constitutive nuclear translocation of TAZ and Hippo pathway repression.
WWTR1 S89W expression was found to lead to acquisition of oncogenic behavior, including increased proliferation, migration, and colony formation in vitro as well as increased tumorigenesis in vivo, which could be reversed by targeted inhibition of the TAZ/YAP1 complex with verteporfin. Finally, xenografts expressing
WWTR1 S89W displayed a shift in tumor phenotype, becoming more infiltrative as well as less differentiated, and were found to be composed of cells with conspicuous cytoplasmic clearing as compared to controls. Our results demonstrate that Hippo pathway alterations are likely drivers of cCCCs and likely contribute to the clear cell phenotype. Therapies targeting this pathway may constitute a new class of treatment for these rare, aggressive tumors. © 2022 The Pathological Society of Great Britain and Ireland.