PARP抑製劑和放射線可增強體外肝細胞死亡。肝細胞癌有跟腱

StephenW

PARP inhibitors and radiation potentiate liver cell death in vitro. Do hepatocellular carcinomas have an achilles' heel?
Laetitia Gerossier  1 , Anaëlle Dubois  1 , Alexia Paturel  1 , Nadim Fares  1 , Damien Cohen  1 , Phillippe Merle  2 , Joel Lachuer  3 , Anne Wierinckx  3 , Pierre Saintigny  4 , Brigitte Bancel  5 , Janick Selves  6 , Anne Schnitzler  7 , Bérengère Ouine  8 , Aurélie Cartier  8 , Leanne de Koning  8 , Vincent Puard  8 , Ivan Bieche  7 , Hector Hernandez-Vargas  1 , Janet Hall  1 , Isabelle Chemin  9
Affiliations
Affiliations

    1
    Univ Lyon, Université Claude Bernard Lyon 1, INSERM, CNRS, Centre Léon Bérard, Centre De Recherche En Cancérologie De Lyon, Lyon, 69008, France.
    2
    Univ Lyon, Université Claude Bernard Lyon 1, INSERM, CNRS, Centre Léon Bérard, Centre De Recherche En Cancérologie De Lyon, Lyon, 69008, France; Department of Hepatology, Groupement Hospitalier Nord, Hospices Civils De Lyon, Lyon, 69000 France.
    3
    Univ Lyon, Université Claude Bernard Lyon 1, INSERM, CNRS, Centre Léon Bérard, Centre De Recherche En Cancérologie De Lyon, Lyon, 69008, France; ProfileXpert, SFR-Est, CNRS UMR-S3453, INSERM US7, Lyon Cedex 08, F-69373, France.
    4
    Univ Lyon, Université Claude Bernard Lyon 1, INSERM, CNRS, Centre Léon Bérard, Centre De Recherche En Cancérologie De Lyon, Lyon, 69008, France; Department of Medical Oncology, Centre Léon Bérard, Lyon, France.
    5
    Service d'Anatomopathologie, Groupement Hospitalier Est, Hospices Civils De Lyon, Lyon, 69000, France.
    6
    Anatomie Et Cytologie Pathologiques Pôle IUC Oncopole CHU Institut Universitaire Du Cancer De Toulouse - Oncopole, Toulouse, F- 31059, France.
    7
    Department of Genetics, Institut Curie, PSL Research University, Paris, F-75005, France.
    8
    Department of Translational Research, Institut Curie, PSL Research University, Paris, F-75005, France.
    9
    Univ Lyon, Université Claude Bernard Lyon 1, INSERM, CNRS, Centre Léon Bérard, Centre De Recherche En Cancérologie De Lyon, Lyon, 69008, France. Electronic address: [email protected].

    PMID: 33183998 DOI: 10.1016/j.clinre.2020.09.014

Abstract

Background: A promising avenue for cancer treatment is exacerbating the deregulation of the DNA repair machinery that would normally protect the genome. To address the applicability of poly(ADP-ribose) polymerase (PARP) inhibitors (PARPi) combined with radiotherapy for the treatment of hepatocellular carcinoma (HCC) two approaches were used: firstly, the in vitro sensitivity to the PARPi Veliparib and Talazoparib +/- radiation exposure was determined in liver cell lines and the impact of the HBV X protein (HBx) that deregulates cellular DNA damage repair via SMC5/6 degradation was investigated. Secondly, PARP expression profiles and DNA damage levels using the surrogate marker gammaH2AX were assessed in a panel of control liver vs HCC tissues.

Methods: Cell cytotoxicity was measured by clonogenic survival or relative cell growth and the DNA damage response using immunological-based techniques in Hep3B, PLC/PRF/5, HepG2- and HepaRG-derived models. Transcriptome changes due to HBx expression vs SMC6 loss were assessed by RNA sequencing in HepaRG-derived models. PARP and PARG transcripts (qPCR) and PARP1, H2AX and gammaH2AX protein levels (RPPA) were compared in control liver vs HBV-, HCV-, alcohol- and non-alcoholic steatohepatitis-associated HCC (tumor/peritumor) tissues.

Results: PARPi cytotoxicity was significantly enhanced when combined with X-rays (2 Gy) with Talazoparib having a greater impact than Veliparib in most in vitro models. HBx expression significantly lowered survival, probably driven by SMC5/6 loss based on the transcriptome analysis and higher DNA damage levels. PARP1 and PARP2 transcript levels were significantly higher in tumor than peritumor and control tissues. The HBV/HCV/alcohol-associated tumor tissues studied had reduced H2AX but higher gammaH2AX protein levels compared to peritumor and control tissues providing evidence of increased DNA damage during liver disease progression.

Conclusions: These proof-of-concept experiments support PARPi alone or combined with radiotherapy for HCC treatment, particularly for HBV-associated tumors, that warrant further investigation.

Keywords: DNA damage; Hepatitis B virus (HBV) X protein; Liver cancer; SMC5/6; Talazoparib; Veliparib.; gammaH2AX.

Copyright © 2020. Published by Elsevier Masson SAS.

StephenW

PARP抑製劑和放射線可增強體外肝細胞死亡。肝細胞癌有跟腱嗎？
Laetitia Gerossier 1，AnaëlleDubois 1，Alexia Paturel 1，Nadim Fares 1，Damien Cohen 1，Phillippe Merle 2，Joel Lachuer 3，Anne Wierinckx 3，Pierre Saintigny 4，Brigitte Bancel 5，Janick Selves 6，Anne Schnitzler 7，BérengèreOu 8，AurélieCartier 8，Leanne de Koning 8，Vincent Puard 8，Ivan Bieche 7，Hector Hernandez-Vargas 1，Janet Hall 1，Isabelle Chemin 9
隸屬關係
隸屬關係

    1個
    里昂大學，克勞德·伯納德大學，里昂1號，INSERM，CNRS，里昂·貝拉爾中心，里昂中心研究中心，里昂，法國69008。
    2
    里昂大學，克勞德·伯納德大學，里昂1號，INSERM，CNRS，里昂·貝拉德中心，里昂中心研究中心，里昂，法國69008；法國里昂醫院，北里昂醫院集團醫院肝病科，法國69000。
    3
    里昂大學，克勞德·伯納德大學，里昂1號，INSERM，CNRS，里昂·貝拉德中心，里昂中心研究中心，里昂，法國69008； ProfileXpert，SFR-Est，CNRS UMR-S3453，INSERM US7，里昂塞德克斯08，F-69373，法國。
    4
    里昂大學，克勞德·伯納德大學，里昂1號，INSERM，CNRS，里昂·貝拉德中心，里昂中心研究中心，里昂，法國69008；法國里昂萊昂貝拉德中心腫瘤內科。
    5
    法國里昂醫院，利比亞收容所Est Anatomopathologie服務部，69000，法國。
    6
    解剖學和細胞學病理學PôleIUC Oncopole CHU大學圖盧茲癌症研究所-Oncopole，圖盧茲，F- 31059，法國。
    7
    PSL研究大學居里研究所遺傳學系，巴黎，F-75005，法國。
    8
    PSL研究大學居里研究所轉化研究系，巴黎，F-75005，法國。
    9
    里昂大學，克勞德·伯納德大學，里昂1號，INSERM，CNRS，里昂·貝拉爾中心，里昂中心研究中心，里昂，法國69008。電子地址：[email protected]。

    PMID：33183998 DOI：10.1016 / j.clinre.2020.09.014

抽象

背景：一種有前途的癌症治療途徑正在加劇通常會保護基因組的DNA修復機制的失控。為了解決聚（ADP-核糖）聚合酶（PARP）抑製劑（PARPi）與放射療法聯合用於治療肝細胞癌（HCC）的適用性，使用了兩種方法：首先，對PARPi Veliparib和Talazoparib的體外敏感性-確定了肝細胞系中的輻射暴露，並研究了通過SMC5 / 6降解來解除細胞DNA損傷修復的HBV X蛋白（HBx）的影響。其次，在一組對照肝臟與肝癌組織中評估了使用替代標誌物gammaH2AX的PARP表達譜和DNA損傷水平。

方法：在Hep3B，PLC / PRF / 5，HepG2-和HepaRG衍生的模型中，通過基於克隆的存活率或相對細胞生長以及基於免疫學的技術測量DNA損傷反應。在HepaRG衍生的模型中，通過RNA測序評估了因HBx表達與SMC6缺失而導致的轉錄組變化。比較了對照肝臟與HBV，HCV，酒精性和非酒精性脂肪性肝炎相關的HCC（腫瘤/腫瘤）組織中的PARP和PARG轉錄本（qPCR）以及PARP1，H2AX和gammaH2AX蛋白水平（RPPA）。

結果：在大多數體外模型中，與Talazoparib聯合X射線（2 Gy）聯合使用時，PARPi的細胞毒性顯著增強，而Talazoparib的影響大於Veliparib。根據轉錄組分析和較高的DNA損傷水平，HBx表達可能會降低SMC5 / 6的表達，從而大大降低生存率。腫瘤中的PARP1和PARP2轉錄水平顯著高於癌旁組織和對照組織。與周圍組織和對照組織相比，所研究的HBV / HCV /酒精相關腫瘤組織的H2AX含量降低，但gammaH2AX蛋白水平更高，從而提供了在肝病進展過程中DNA損傷增加的證據。

結論：這些概念驗證實驗單獨支持PARPi或與放療聯合用於HCC治療，特別是用於HBV相關腫瘤，值得進一步研究。

關鍵詞：DNA損傷乙型肝炎病毒（HBV）X蛋白；肝癌; SMC5 / 6;塔拉唑帕尼; Veliparib。 gammaH2AX。

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