Tumor-selective use of PARP inhibitors against NQO1+ nonsmall cell lung cancer

我们的长期目标是开发新的抗肿瘤疗法来治疗顽固性非小细胞肺癌。 (NSCLC)通过利用其独特的双电子第二相解毒酶的过度表达， NAD(P)H：苯醌氧化还原酶1(NQO1)与聚腺苷二磷酸核糖抑制剂的联合应用 聚合酶抑制剂。在非小细胞肺癌中，NQO1比相应的正常组织升高5-200倍， 而过氧化氢酶在非小细胞肺癌中的表达水平低于相关的正常肺组织 (或所有其他正常)组织。我们证明了独特和新颖的NQO1生物活性药物(例如 拉帕酮(临床试验中的ARQ761)可被NQO1激活，导致大量的过氧化氢诱导， 脱嘧啶/无嘌呤(AP)位点和DNA单链断裂介导的聚(ADP- 核糖基)聚合酶1(PARP1)，导致NAD+的显著损失(Huang等人，癌症资源，2012年)。 然而，联合应用PARP抑制剂可显著增强这些药物的抗肿瘤活性。 药物(Huang等人，癌症细胞，2016)。我们发现了一种效力更强的NQO1生物活性物质 异丁基脱氧苯二酚(IB-DNQ)与抑制PARP的协同作用显著增强， 伯拉帕乔尼。我们假设IB-DNQ单独使用会比ARQ761(？) 雷帕酮)，并显著增强NQO1依赖的协同抗肿瘤活性 联合PARP抑制剂对NQO1+NSCLC细胞的抑制作用。我们建议完成三个具体的 目的1：明确PARP抑制+IB-DNQ的作用机制 协同细胞死亡(年0-5年)；目标2：确定IB-DNQ诱导的过氧化氢相关的作用 NQO1+程序性坏死与NQO1-细胞凋亡同时发生的旁观者效应 NSCLC细胞(0-5年)；和目标3：定义肿瘤选择性的、NQO1+依赖的抗肿瘤 PARP抑制+IB-DNQ方案与单纯IB-DNQ方案在非小细胞肺癌原位移植中的作用 (0-5岁)。AIMS将使用基因匹配的NQO1+与NQO1-NSCLC细胞系通过 抑制NQO1+人癌细胞株的SI/shRNA和/或强迫NQO1过度表达匹配 多态NQO1-人类NSCLC中的已知水平。我们还将检查PARP1的基因匹配情况 SI/shRNA基因敲除与正常非小细胞肺癌细胞的体内外比较。 我们组建了一支优秀的研究团队来探索新陈代谢、旁观者和疗效的影响 在这项为期5年的研究中，PARP抑制+IB-DNQ治疗用于原位NSCLC异种移植模型。我们的 结果应该会在5年内导致使用这种组合的临床试验的发展。

Our long-term goal is to develop novel antitumor therapies to treat recalcitrant non-small cell lung cancers (NSCLCs) by exploiting their unique over-expression of the two-electron, Phase II detoxifying enzyme, NAD(P)H:Quinone Oxidoreductase 1 (NQO1) in combination with inhibitors of poly(ADP-ribose) polymerase inhibitors. In NSCLC, NQO1 is elevated 5- to 200-fold above associated normal tissues, while catalase levels are expressed at extremely lower levels in NSCLCs than in associated normal lung (or all other normal) tissue. We showed that unique and novel NQO1 bioactivatable drugs, (e.g., ß- lapachone (ARQ761 in clinical trials), are `bioactivated' by NQO1, resulting in massive H2O2-induced, apyrimidinic/apurinic (AP) site- and DNA single-strand break-mediated hyperactivation of poly(ADP- ribosyl) polymerase 1 (PARP1) that causes dramatic NAD+ losses (Huang et al., Cancer Res, 2012). However, co-administration of a PARP inhibitor dramatically enhances the antitumor activities of these drugs (Huang et al., Cancer Cell, 2016). We have discovered a much more potent NQO1 bioactivatable drug, isobutyldeoxynyboquinone (IB-DNQ) that is significantly more synergistic with PARP inhibition that ß-lapachone. We hypothesize that IB-DNQ will be a significantly better drug alone than ARQ761 (ß- lapachone) and result in significantly greater synergistic, NQO1-dependent antitumor activity in combination with PARP inhibitors against NQO1+ NSCLC cells. We propose to complete three specific aims to test this hypothesis.: Aim 1: To define the mechanism of action of PARP inhibition + IB-DNQ synergistic cell death (Yrs 0-5); Aim 2: To define the role of IB-DNQ-induced H2O2-related bystander effect in simultaneous NQO1+ programmed necrosis vs NQO1- apoptosis in matched NSCLC cells (Yrs 0-5); and Aim 3: To define the tumor-selective, NQO1+-dependent antitumor effects of the PARP inhibition + IB-DNQ regimen vs IB-DNQ alone in orthotopic NSCLC xenografts (Yrs 0-5). The aims will be performed using genetically matched NQO1+ vs NQO1- NSCLC cell lines by si/shRNA knockdown of NQO1+ human cancer cell lines and/or forced NQO1 over-expression to match known levels in polymorphic NQO1- human NSCLCs. We will also examine genetically match PARP1 si/shRNA knockdown vs normal NSCLC cells in vitro and in vivo. We assembled an outstanding research team to explore the metabolic, bystander, and efficacy effects of PARP inhibition + IB-DNQ treatments in orthotopic NSCLC xenograft models in this 5-yr grant. Our results should lead to the development of a clinical trial using this combination in 5 years.