Next-generation sequencing of advanced prostate cancer treated with androgen-deprivation therapy.
Next-generation sequencing of advanced prostate cancer treated with androgen-deprivation therapy.
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DOI：
10.1016/j.eururo.2013.08.011
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发表时间：
2014-07
期刊：
影响因子：
23.4
通讯作者：
Leung, Hing Y.
中科院分区：
文献类型：
--
作者：
Rajan, Prabhakar;Sudbery, Ian M.;Villasevil, M. Eugenia M.;Mui, Ernest;Fleming, Janis;Davis, Mark;Ahmad, Imran;Edwards, Joanne;Sansom, Owen J.;Sims, David;Ponting, Chris P.;Heger, Andreas;McMenemin, Rhona M.;Pedley, Ian D.;Leung, Hing Y.
雄激素剥夺疗法（ADT）是局部晚期或转移性前列腺癌（PCa）的标准治疗。许多患者在大约2-3年后出现去势抵抗（castration-resistant PCa [CRPC]），预后较差。CRPC进展的分子机制尚不清楚。在ADT之前和之后进行定量肿瘤转录组分析，以确定功能重要的雄激素调节途径或可能在CRPC中重新激活的基因。RNA测序（RNA-seq）对7例局部晚期或转移性前列腺癌患者在ADT开始前和大约22周后进行了富含肿瘤的靶向前列腺活检。在治疗对中鉴定出差异调节基因，并通过细胞系上的定量逆转录-聚合酶链反应（qRT-PCR）和单独的CRPC患者队列的免疫组织化学进一步研究。功能分析用于确定通路调节对细胞表型的影响。我们寻找影响关键细胞信号通路的基因表达变化，这些变化可能作为CRPC体外细胞系模型的原理证明。我们确定了adt调节的信号通路，包括Wnt/β-catenin信号通路，并通过免疫组织化学观察到β-catenin在CRPC亚群中的过表达。我们在LNCaP/LNCaP- ai细胞rna上通过qRT-PCR验证了12个通路成员中的6个（50%），其中4个（67%）的表达变化与RNA-seq数据一致。我们发现，与雄激素响应型LNCaP细胞相比，tankyrase抑制剂XAV939（促进β-catenin降解）通过G0/G1期细胞比例的积累和S期和G2/M期细胞比例的减少，降低了雄激素非依赖性LNCaP- ai细胞系的生长。我们的活检方案没有考虑到肿瘤的异质性，途径抑制仅限于药理学方法。配对PCa样本的rna测序显示adt调控的信号通路。Wnt/β-catenin信号通路特异性延迟雄激素非依赖性PCa细胞周期进展和增殖的原理证明，值得进一步研究作为CRPC治疗的潜在靶点。在雄激素剥夺治疗期间，对前列腺癌（PCa）转录组的全面RNA测序分析确定了几种细胞信号通路中的基因表达变化，包括Wnt/β-catenin信号通路，该通路在雄激素非依赖性PCa细胞生长中具有潜在作用。
Androgen-deprivation therapy (ADT) is standard treatment for locally advanced or metastatic prostate cancer (PCa). Many patients develop castration resistance (castration-resistant PCa [CRPC]) after approximately 2–3 yr, with a poor prognosis. The molecular mechanisms underlying CRPC progression are unclear. To undertake quantitative tumour transcriptome profiling prior to and following ADT to identify functionally important androgen-regulated pathways or genes that may be reactivated in CRPC. RNA sequencing (RNA-seq) was performed on tumour-rich, targeted prostatic biopsies from seven patients with locally advanced or metastatic PCa before and approximately 22 wk after ADT initiation. Differentially regulated genes were identified in treatment pairs and further investigated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) on cell lines and immunohistochemistry on a separate CRPC patient cohort. Functional assays were used to determine the effect of pathway modulation on cell phenotypes. We searched for gene expression changes affecting key cell signalling pathways that may be targeted as proof of principle in a CRPC in vitro cell line model. We identified ADT-regulated signalling pathways, including the Wnt/β-catenin signalling pathway, and observed overexpression of β-catenin in a subset of CRPC by immunohistochemistry. We validated 6 of 12 (50%) pathway members by qRT-PCR on LNCaP/LNCaP-AI cell RNAs, of which 4 (67%) demonstrated expression changes consistent with RNA-seq data. We show that the tankyrase inhibitor XAV939 (which promotes β-catenin degradation) reduced androgen-independent LNCaP-AI cell line growth compared with androgen-responsive LNCaP cells via an accumulation of cell proportions in the G0/G1 phase and reduction in the S and G2/M phases. Our biopsy protocol did not account for tumour heterogeneity, and pathway inhibition was limited to pharmacologic approaches. RNA-seq of paired PCa samples revealed ADT-regulated signalling pathways. Proof-of-principle inhibition of the Wnt/β-catenin signalling pathway specifically delays androgen-independent PCa cell cycle progression and proliferation and warrants further investigation as a potential target for therapy for CRPC. A comprehensive RNA sequencing analysis of the prostate cancer (PCa) transcriptome during androgen-deprivation therapy identifies gene expression changes within several cell-signalling pathways, including the Wnt/β-catenin signalling pathway, which has a potential role in androgen-independent PCa cell growth.
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影响因子：
3.7
作者：
Rajan, Prabhakar;Dalgliesh, Caroline;Elliott, David J.
通讯作者：
Elliott, David J.
影响因子：
158.5
作者：
Scher, Howard I.;Fizazi, Karim;de Bono, Johann S.
通讯作者：
de Bono, Johann S.
影响因子：
50.3
作者：
Sharma, Naomi L.;Massie, Charlie E.;Neal, David E.
通讯作者：
Neal, David E.
影响因子：
14.9
作者：
't Hoen PA;Ariyurek Y;Thygesen HH;Vreugdenhil E;Vossen RH;de Menezes RX;Boer JM;van Ommen GJ;den Dunnen JT
通讯作者：
den Dunnen JT
影响因子：
50.3
作者：
Carver BS;Chapinski C;Wongvipat J;Hieronymus H;Chen Y;Chandarlapaty S;Arora VK;Le C;Koutcher J;Scher H;Scardino PT;Rosen N;Sawyers CL
通讯作者：
Sawyers CL