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

项目2 顺铂（CDDP）是人类乳头状瘤病毒（HPV）阳性和阴性的一线治疗药物 和颈部鳞状细胞癌（HNSCC）。不幸的是，没有可重复的CDDP预测因子 HNSCC反应和患者发展获得性耐药，甚至在一个疗程的化疗后发生远处转移。 这通常是有毒的化疗。为了更好地理解和解决CDDP治疗失败的临床问题， 我们使用已建立的HNSCC在实验室中开发了获得性CDDP抗性的多种细胞系模型 肿瘤细胞系具有不同的基因组背景和TP53突变和功能状态。令人惊讶的是，几乎 从这些不同的基因组驱动背景中开发的每一个CDDP抗性克隆都具有CDDP的超活化。 Nrf2途径是由于KEAP 1的从头体细胞突变或RNA表达降低（阴性 Nrf2）。nrf2激活与预后不良、对免疫治疗缺乏反应、 转移和化疗抗性。我们证明了Nrf2的过度激活是必要的， 维持获得性耐药性，提高了该途径在临床上被靶向逆转或 预防HNSCC中获得性CDDP耐药性。通过用一种新型药物靶向谷氨酰胺酶1（GLS）， Nrf2活性所需的谷胱甘肽被耗尽，导致CDDP抗性逆转。 与此同时，我们发现感觉神经元之间的旁分泌信号在大脑中被重新编程， 肿瘤微环境可以促进CDDP抗性， 该信号轴可增强CDDP敏感性和肿瘤进展。此外，我们的临床前模型的数据 这表明伴随CDDP抗性Nrf2活化也可能导致更具侵袭性的疾病， 增加远处转移的几率。虽然Nrf2过度激活可能是一个允许的和必要的步骤， 获得性CDDP抗性，也可能存在Nrf2独立途径促成抗性。我们的中央 假设是Nrf2过度激活通过体细胞突变进行转录控制是常见的， 在一系列基因组背景下，CDDP获得性耐药性的发展中存在靶向事件。到 弥合Nrf2通路在CDDP耐药性和肿瘤发展中的普遍性方面的知识空白 进展和克服治疗失败的方法，我们提出以下目的：1）定义贡献 HNSCC中获得性CDDP耐药的Nrf2依赖性和非依赖性途径; 2）描述HNSCC中获得性CDDP耐药的Nrf2依赖性和非依赖性途径; 获得性CDDP抗性、Nrf2激活、神经元 3）确定是否获得性CDDP抗性和CDDP抗性是否在肿瘤微环境中发生。 肿瘤进展可以通过靶向代谢或稳定p53的药物来克服。完成这些 目的应该产生可翻译的结果，这将改善HNSCC患者的治疗和相关疾病。 上呼吸消化道肺癌和食道癌，因此对癌症有重大影响 国际吧

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.