Overcoming Hypoxic Resistance in Non-Small Cell Lung Cancer By Targeting Mitochondrial Metabolism

项目摘要 许多研究小组正在研究为什么一些肺癌患者对放射治疗和免疫治疗反应良好， 有些则不然。一个变量是肿瘤缺氧，许多研究小组已经表明它可以显着抑制肿瘤生长。 这些治疗方法的有效性。临床研究已经确定缺氧是一种独立的 患者预后不良的预后指标，但即使这种联系已经知道了几十年，没有 FDA批准的干预措施可以在临床上克服缺氧。一些调查人员试图提供更多 氧气，但这种方法仍然受到限制，由于不良形成的肿瘤脉管系统。我们 采取了一种创新的方法，并询问我们是否可以减少需求，而不是增加供应， 氧气减少缺氧。我们发现FDA批准的血管舒张剂罂粟碱（PPV）具有关闭- 以低微摩尔浓度靶向抑制线粒体复合物1并迅速降低氧消耗的能力 在体外测试的每个细胞系中的浓度。我们还表明，PPV可以提高有效性， 在肺癌和其他癌症的临床前模型中进行放射和免疫检查点阻断（ICB）， 使氧合良好的正常组织敏感。减少缺氧逆转免疫赦免，减少终末- 耗尽的T细胞，并增加对PD-1阻断有反应的祖细胞。我们最近有更多的 开发了PPV的新衍生物，这些衍生物失去了血管舒张能力，并增加了其持续时间。 作用，使它们可以改善免疫增敏剂。我们现在提出检验PPV可以 有效增强肺癌临床前模型的放射和免疫治疗， 在晚期非小细胞肺癌（NSCLC）标准治疗中增加PPV是可行的。我们有 研究了TCGA数据库，发现KEAP 1/NRF 2通路中的肺癌驱动突变导致了 高水平的线粒体基因表达可导致氧代谢升高，从而导致缺氧。 在目的1中，我们将研究致癌NRF 2激活人类和小鼠细胞和模型肿瘤的作用 为了确定这些细胞对线粒体功能的依赖性， 导致肿瘤缺氧，以及治疗难治性肿瘤是否被PPV或其衍生物致敏。在目标2中， 我们将研究肿瘤缺氧对模型肿瘤中T细胞迁移和活化的影响，以及如何影响T细胞的迁移和活化。 PPV或其衍生物减少缺氧后，免疫浸润发生变化。最后，在目标3中， 进行1期临床试验，以确定接受标准PPV治疗的患者是否可行 晚期非小细胞肺癌的护理放化疗后免疫治疗。我们将寻求改变的有效性 使用配对血液水平氧测定（BOLD）MRI的肿瘤氧合，以及免疫组织化学的变化。 外周血单核细胞群。这些研究将让我们知道是否以及如何使用PPV或其 在未来的临床试验中用于治疗NSCLC的新型衍生物。

PROJECT SUMMARY Many groups are investigating why some lung cancer patients respond well to radio- and immuno-therapies and some do not. One variable is tumor hypoxia, and many groups have shown it can significantly inhibit the effectiveness to these therapeutic modalities. Clinical studies have identified hypoxia as an independent prognostic indicator of poor patient outcomes, but even though this connection has been known for decades, no FDA-approved intervention exists to clinically overcome hypoxia. Some investigators have tried to deliver more oxygen to the tumor, but this approach remains constrained due to the poorly formed tumor vasculature. We have taken an innovative approach and asked if we can reduce demand for, rather than increase supply of, oxygen to reduce hypoxia. We have found that the FDA-approved vasorelaxant papaverine (PPV) has an off- target ability to inhibit mitochondrial complex 1, and reduce oxygen consumption rapidly, in low micromolar concentrations in every cell line tested in vitro. We have also shown that PPV can enhance the effectiveness of radiation and immune checkpoint blockade (ICB) in preclinical models of lung and other cancers, without sensitizing well-oxygenated normal tissue. Reducing hypoxia reverses immune privilege, decreases terminally- exhausted T cells, and increases progenitors that are responsive to PD-1 blockade. We have more recently developed new derivatives of PPV that have lost their vasorelaxant capability and increased their duration of action so that they can be improved immuno-sensitizers. We now propose to test the hypothesis that PPV can effectively enhance the radio- and immuno-therapeutic treatment of preclinical models of lung cancer, and that it is feasible to add PPV to standard of care therapy for advanced non-small cell lung cancer (NSCLC). We have examined TCGA databases and found that lung cancer driver mutations in the KEAP1/NRF2 pathway lead to high levels of mitochondrial gene expression that can cause elevated oxygen metabolism contributing to hypoxia. In Aim 1, we will investigate the effects of oncogenic NRF2 activation human and murine cells and model tumors to determine the dependence of these cells on mitochondrial function, how increased oxygen metabolism contributes to tumor hypoxia, and if therapy-refractory tumors are sensitized by PPV or its derivatives. In Aim 2, we will examine the effect of tumor hypoxia on the migration and activation of T-cells in model tumors and how the immune infiltrate changes after reduction of hypoxia with PPV or its derivatives. Finally, in Aim 3 we will perform a phase 1 clinical trial to determine if the addition of PPV is feasible for patients receiving standard of care chemoradiation followed by immunotherapy for advanced NSCLC. We will look for effectiveness in changing tumor oxygenation using paired blood level oxygen determination (BOLD) MRIs, and for changes in immune populations of peripheral blood mononuclear cells. These studies will let us know if, and how, to use PPV or its novel derivatives in future clinical trials for the treatment of NSCLC.