Tumor-selective radiosensitization of NSCLC using NQO1 bioactivatable drugs

Boothman，DA 摘要 在非小细胞肺癌（NSCLC）中，>80%的肿瘤NQO 1水平升高5- 200倍 高于相关的正常组织，而过氧化氢酶水平表达在极低的水平， NSCLC与体内所有正常组织相比。我们发现，独特而新颖的NQO 1 可生物活化的药物，β-拉帕酮（ARQ 761在临床试验中）和一种新药， 异丁基脱氧尼波醌（IB-DNQ）被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poly（ADP- 核糖基）聚合酶1（PARP 1），其导致显著的NAD+/ATP损失。拉帕乔内（Lapachone）也是 对过表达NQO 1的癌细胞具有良好的放射增敏作用。我们假设， 低剂量电离辐射（IR）的NQO 1 + NSCLC癌症，随后立即使用无毒 NQO 1生物可活化药物（拉帕醌或IB-DNQ 2小时）将导致DNA水平升高 在过度表达NQO 1的细胞中过度激活PARP活性的病变。低剂量的IR或抗- 拉帕醌不能单独引起显著水平的DNA损伤而过度激活PARP， 将一起使用。由于IR会导致快速修复的DSB，因此必须交付该代理 在NQO 1生物活化药物之前或同时，18，19然后必须应用至少2 最大DNA损伤形成、PARP过度活化和代谢改变的时间为1.8 h， DSB修复并导致NQO 1 + NSCLC细胞的协同致死。将实现三个目标： 目的1：确定DNA修复抑制作用（对同源重组（HR）和非同源重组（NHR））， 同源末端连接（NHEJ））。(Yrs 0-5）。预防：非致死，低剂量 IR或β-lap可用于引起DSB和PARP过度激活，从而耗尽NAD+/ATP水平 并抑制DSB修复（NHEJ和HR）以增强对NSCLC的功效。 目的2：确定碳代谢的抑制（糖酵解和TCA循环抑制，由于 NAD+/ATP损失）。假设：PARP过度激活导致戏剧性的 NAD+/ATP损失，极大地抑制了糖酵解（通过GAPDH抑制）和LDH引起的TCA循环 抑制和其他NAD（P）H依赖性途径，以最大限度地提高对NQO 1 + NSCLC的疗效。 目的3：阐明IR + NQO 1生物可活化药物组合的最有效用途 针对NQO 1 + NSCLC。(Yrs 0-5）。前提：IR + NQO 1生物活化药物会诱导DSB + 过度激活PARP的DNA损伤，导致NAD+/ATP的急剧损失，抑制DSB修复，大大 抑制诱导肿瘤选择性程序性坏死的糖酵解和TCA循环代谢。 一个杰出的研究团队将探索IR + NQO 1生物活化药物对DSB的影响 修复，代谢和对NSCLC异种移植模型的疗效，导致3-5年的临床试验。

Boothman, DA Abstract In non-small cell lung cancer (NSCLC), >80% tumors have elevated NQO1 levels 5- to 200-fold above associated normal tissues, while Catalase levels are expressed at extremely low levels in NSCLCs compared to all normal tissue in the body. We showed that unique and novel NQO1 bioactivatable drugs, ß-lapachone (ARQ761 in clinical trials) and a new drug, isobutyldeoxynyboquinone (IB-DNQ), 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+/ATP losses. ß-Lapachone is also an excellent radiosensitizer against cancer cells overexpressing NQO1. We hypothesize that exposure of NQO1+ NSCLC cancers with low dose ionizing radiation (IR), followed immediately with nontoxic NQO1 bioactivatable drugs (ß-lapachone or IB-DNQ for 2 h) will cause an elevated level of DNA lesions that hyperactivate PARP activity in cells that over-express NQO1. Low doses of IR or ß- lapachone, which are not able alone to cause a significant level of DNA lesions to hyperactivate PARP, will be used together. Since IR causes DSBs that are quickly repaired, this agent must be delivered before or at the same time as NQO1 bioactivatable drugs,18,19 which then must be applied for at least 2 h for maximal DNA lesion formation, PARP hyperactivation and metabolic alterations7,8,12 that inhibit DSB repair and cause synergistic lethality of NQO1+ NSCLC cells. Three aims will be performed: Aim 1: To define DNA repair inhibitory effects (on homologous recombination (HR) and non- homologous end joining (NHEJ)) after IR + ß-lapachone. (Yrs 0-5). Premise: Nonlethal, low doses of IR or ß-lap can be used to cause DSBs and PARP hyperactivation that depletes NAD+/ATP levels and inhibits DSB repair (NHEJ and HR) to enhance efficacy against NSCLC. Aim 2: To define the inhibition of carbon metabolism (glycolytic and TCA cycle inhibition due to NAD+/ATP losses) after IR + ß-lapachone (Yrs 0-5). Premise: PARP hyperactivation causes dramatic NAD+/ATP losses that greatly suppress glycolysis (via GAPDH inhibition) and TCA cycling by LDH inhibition and other NAD(P)H-dependent pathways to maximize efficacy against NQO1+ NSCLC. Aim 3: To elucidate the most efficacious use of IR + NQO1 bioactivatable drug combination against NQO1+ NSCLCs. (Yrs 0-5). Premise: IR + NQO1 bioactivatable drugs will induce DSBs + DNA lesions that hyperactivate PARP, lead to dramatic NAD+/ATP losses, inhibit DSB repair, greatly suppress glycolysis and TCA cycle metabolism that induces tumor-selective programmed necrosis. An outstanding research team will explore the effects of IR + NQO1 bioactivatable drugs on DSB repair, metabolism and efficacy against NSCLC xenograft models, leading to a clinical trial in 3-5 years.