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复发性非小细胞肺癌 （NSCLC）通过利用其独特的双电子II相解毒酶的过表达， NAD（P）H：醌氧化还原酶1（NQO 1）与聚（ADP-核糖）抑制剂的组合 聚合酶抑制剂在NSCLC中，NQO 1比相关正常组织升高5- 200倍， 而过氧化氢酶水平在NSCLC中的表达水平比在相关的正常肺中的表达水平低得多， (or所有其它正常）组织。我们表明，独特的和新的NQO 1生物活化药物，（例如，- 拉帕醌（ARQ 761在临床试验中），是由NQO 1“生物活化”，导致大量的H2 O2诱导， 无嘧啶/无嘌呤（AP）位点和DNA单链断裂介导的聚（ADP-）超活化 核糖基）聚合酶1（PARP 1）引起显著的NAD+损失（Huang等人，Cancer Res，2012）。 然而，PARP抑制剂的共同施用显著增强了这些化合物的抗肿瘤活性。 药物（Huang等人，Cancer Cell，2016）。我们已经发现了一种更有效的NQO 1生物活化 药物，异丁基脱氧尼波醌（IB-DNQ），它与PARP抑制作用具有显著的协同作用， - 拉帕乔内我们假设IB-DNQ将是一种比ARQ 761更好的单独药物（2015年）。 拉帕酮），并导致显著更大协同性、NQO 1依赖性抗肿瘤活性， 与PARP抑制剂联合治疗NQO 1 + NSCLC细胞。我们建议完成三个具体的 旨在验证这一假设。目的1：确定PARP抑制+ IB-DNQ的作用机制 目的2：确定IB-DNQ诱导的H2 O2相关细胞死亡的作用。 同时发生NQO 1+程序性坏死与NQO 1-凋亡的旁观者效应 目的3：确定肿瘤选择性、NQO 1+依赖性抗肿瘤药物 PARP抑制+ IB-DNQ方案与IB-DNQ单独给药在原位NSCLC异种移植物中的作用 (Yrs 0-5）。将使用基因匹配的NQO 1 + vs NQO 1- NSCLC细胞系，通过 NQO 1+人癌细胞系的si/shRNA敲低和/或迫使NQO 1过表达以匹配 多态性NQO 1-人NSCLC中的已知水平。我们还将检查基因匹配PARP 1 在体外和体内与正常NSCLC细胞相比的si/shRNA敲低。 我们组建了一个优秀的研究团队，以探索代谢，旁观者，和疗效的影响， PARP抑制+ IB-DNQ治疗原位NSCLC异种移植模型，为期5年。我们 结果应导致在5年内开发使用该组合的临床试验。

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.