Defining the role of KEAP/NRF2 signaling dysregulation and sensory nerve reprograming during acquisition of cisplatin resistance and metastasis in HNSCC

项目2总结 顺铂(CDDP)是治疗HPV阳性和阴性头部的一线药物 和颈部鳞状细胞癌(HNSCC)。不幸的是，CDDP没有可重复的预测因子。 HNSCC反应和患者在经过一个疗程后出现获得性耐药甚至远处转移 这种化疗往往是有毒的。为了更好地了解和解决顺铂治疗失败的临床问题， 我们在实验室中利用已建立的HNSCC建立了获得性顺铂耐药的多个细胞系模型 具有不同基因组背景和TP53突变和功能状态的肿瘤株。令人惊讶的是，几乎 从这些不同的基因组驱动背景发展出来的每个抗顺铂的克隆都具有高度激活的 Nrf2途径要么是由于Keap1的从头体细胞突变，要么是由于Keap1的RNA表达减少(阴性 NRF2的调节器)。NRF2的激活与预后不良、对免疫治疗缺乏反应有关， 转移和化疗耐药。我们证明了Nrf2的过度激活是必要的 维持获得性耐药，增加了该通路可能被临床靶向逆转或 预防HNSCC获得性顺铂耐药。通过用一种新药靶向谷氨酰胺酶1(GLS)， Nrf2活性所需的谷胱甘肽被耗尽，导致CDDP耐药性逆转。 同时，我们证明了感觉神经元之间的旁分泌信号变得重新编程 肿瘤微环境可促进顺铂耐药，并有可能在药物上抑制 这一信号轴促进了CDDP的敏感性和肿瘤进展。此外，我们临床前模型的数据 提示伴随CDDP耐药的Nrf2激活也可能通过以下方式导致更具侵袭性的疾病 增加远处转移率。虽然NRF2过度激活可能是一个允许和必需的步骤 对于获得性顺铂耐药，也可能存在Nrf2独立途径参与耐药。我们的中央 假设Nrf2的过度激活通过体细胞突变来进行转录控制是一种常见的 在一系列基因组背景的获得性顺铂耐药性的发展中发生有针对性的事件。至 弥合Nrf2通路在顺铂耐药和肿瘤发展中渗透性方面的知识空白 进展和克服治疗失败的方法我们提出了以下目标：1)确定贡献 Nrf2依赖和非依赖途径在HNSCC中获得顺铂耐药；2)描绘 获得性CDDP抵抗、Nrf2激活、神经元之间的功能和机械相互作用 在肿瘤微环境中重新编程，以及DM；3)确定获得性顺铂耐药性和 肿瘤进展可以通过靶向新陈代谢或稳定p53的药物来克服。实现这些目标 AIMS应产生可翻译的结果，以改善HNSCC患者和相关患者的治疗 上呼吸道肺癌和食道癌，因此对癌症有重大影响 全世界。

Project 2 SUMMARY Cisplatin (CDDP) is frontline therapy for both Human Papilloma Virus (HPV) positive and negative head and neck squamous cell carcinomas (HNSCC). Unfortunately, there are no reproducible predictors of CDDP HNSCC response and patients develop acquired resistance and even distant metastasis following a course of this often toxic chemotherapy. To better understand and tackle the clinical problem of CDDP treatment failure, we developed multiple cell line models of acquired CDDP resistance in the laboratory using established HNSCC tumor lines with different genomic backgrounds and TP53 mutational and functional status. Surprisingly, nearly every CDDP resistant clone developed from these various genomic driver backgrounds had hyperactivation of the Nrf2 pathway either due to de novo somatic mutations in, or decreased RNA expression of, KEAP1 (negative regulator of Nrf2). Nrf2 activation has been linked to poor prognosis, lack of response to immune therapy, metastasis, and chemotherapy resistance. We demonstrated that Nrf2 hyperactivation was necessary to maintain acquired resistance, raising the possibility that the pathway could be clinically targeted to reverse or prevent acquired CDDP resistance in HNSCC. By targeting glutaminase 1 (GLS) with a novel drug, pools of glutathione needed for Nrf2 activity are depleted leading to a reversal of CDDP resistance. In parallel, we showed that paracrine signaling between sensory neurons that become reprogramed in the tumor microenvironment can foster CDDP resistance and that it may be possible to pharmacologically inhibit this signaling axis to boost CDDP sensitivity and tumor progression. Furthermore, data from our preclinical model suggest that Nrf2 activation accompanying CDDP resistance may also lead to more aggressive disease by increasing the rate of distant metastasis. While Nrf2 hyperactivation is likely a permissive and required step to acquired CDDP resistance, there may also be Nrf2 independent pathways contributing to resistance. Our central hypothesis is that Nrf2 hyperactivation either through somatic mutation for transcriptional control is a common and targetable event in development of acquired CDDP resistance across a range of genomic backgrounds. To bridge knowledge gaps in the pervasiveness of the Nrf2 pathway in development of CDDP resistance and tumor progression and ways to overcome treatment failure we propose the following Aims: 1) Define the contribution of Nrf2-dependent and-independent pathways to acquired CDDP resistance in HNSCC; 2) Delineate the functional and mechanistic interactions between acquired CDDP resistance, Nrf2 activation, neuronal reprograming in the tumor microenvironment, and DM; 3) Determine whether acquired CDDP resistance and tumor progression can be overcome by drugs targeting metabolism or stabilizing p53. Accomplishment of these Aims should yield translatable findings that will improve the treatment of patients with HNSCC and the related upper-aerodigestive tract cancers of the lung and esophagus and therefore have a major impact on cancer worldwide.