In Vivo Imaging of Mitochondria Structure and Function in Therapy Resistant Lung Tumors

摘要 这项研究的首要目标是确定有效的代谢为基础的诊断和治疗策略 提高非小细胞肺癌（NSCLC）患者的总生存率。我们建议调查 正电子发射断层扫描（PET）示踪剂18F-BnTP作为一种新型代谢诊断剂， 靶向治疗耐药的线粒体氧化代谢的基于代谢的治疗策略 KRAS/LKB 1和EGFR突变型肺肿瘤。2021年，NSCLC将在美国夺走约13万人的生命。肺 肿瘤通常具有高突变负荷，通常使得单药治疗靶向于 致癌驱动突变不成功。此外，肺腺癌的代谢活性亚群 携带KRAS和LKB 1或EGFR突变的LUAD通常对免疫治疗具有耐药性 接近。然而，无论最初从检查点抑制剂或靶向治疗中获益， 大多数患者最终会对治疗产生耐药性。我们用不同的方法 克服NSCLC中的治疗耐药性-即通过其代谢特征对肿瘤进行分类。 在这里，肿瘤是根据它们的代谢依赖性而不是仅仅根据它们的遗传特性来分组和靶向的。 改变。NSCLC是一种代谢异质性疾病，肿瘤利用糖酵解和氧化代谢 线粒体代谢来生长。线粒体是细胞生物能量学和氧化代谢的场所。 磷酸化（OXPHOS），并且对于肺肿瘤的起始和维持是必需的。由于缺乏在 体内成像探针在生理和机械水平上，我们的知识存在差距， 线粒体生物能量学在NSCLC中受到调节。为了解决这一差距，我们功能性地想象了 利用PET成像示踪剂18F-BnTP研究肺肿瘤中的线粒体活性，并证明 作为线粒体膜电位（Δ P）和氧化磷酸化的体内生物标志物， （OXPHOS）在肺肿瘤中的作用3.重要的是，通过使用18F-BnTP PET成像，我们能够区分 OXPHOS依赖性和非依赖性肺肿瘤。在治疗上，我们已经证明18F-BnTP 阳性、OXPHOS依赖性LUAD对线粒体复合物I抑制剂敏感。我们假设 18F-BnTP PET成像可用于功能性地描绘线粒体生物能量学和适应性氧化代谢。 因此，本发明提供了在治疗抗性肺肿瘤中的代谢的新方法，以指导用OXPHOS抑制剂的治疗。在目标1中， 在治疗抗性LUAD中进行线粒体生物能量学的体内解剖。在目标2中， 在治疗抵抗性肿瘤中进行适应性氧化代谢的结构和功能体内分析， KRAS/LKB 1和EGFR突变LUAD。在目标3中，我们将纵向描述LUAD中的氧化代谢 晚期疾病患者。拟议的工作与人类健康有关，我们建议， 18F-BnTP PET成像引导的靶向和氧化代谢代表了一种新的治疗策略， 克服KRAS/LKB 1和EGFR突变肿瘤患者的治疗耐药性。

Abstract The overarching goal of this study is to identify effective metabolic based diagnostic and therapeutic strategies to improve the overall survival of patients with Non-small cell lung cancer (NSCLC). We propose to investigate the positron emission tomography (PET) tracer, 18F-BnTP as a novel metabolic diagnostic and to develop metabolic based therapeutic strategies targeting oxidative mitochondrial metabolism in therapy resistant KRAS/LKB1 and EGFR mutant lung tumors. NSCLC will claim the lives of ~130,000 in the US in 2021. Lung tumors frequently possess a high mutational burden, often rendering single agent therapies targeting oncogenic driver mutations unsuccessful. Furthermore, metabolically active subsets of lung adenocarcinomas (LUADs) bearing mutations in KRAS and LKB1 or EGFR are frequently resistant to immunotherapy approaches. However, regardless of the initials benefits from checkpoint inhibitors or targeted therapies, the majority of patients will eventually develop resistance to therapy. We rationalize a different approach to overcoming therapy resistance in NSCLC – namely the classification of tumors by their metabolic signature. Here, tumors are grouped and targeted by their metabolic dependencies rather than solely by their genetic alterations. NSCLC is a metabolically heterogeneous disease and tumors utilize both glycolytic and oxidative mitochondrial metabolism to grow. The mitochondria are the site of cellular bioenergetics and oxidative phosphorylation (OXPHOS) and are essential for lung tumor initiation and maintenance. Due to a lack of in vivo imaging probes there is a gap in our knowledge at a physiological and mechanistic level of how mitochondrial bioenergetics are regulated in NSCLC. To address this gap, we functionally imaged mitochondrial activity in lung tumors utilizing the PET imaging tracer 18F-BnTP and demonstrate that it functions an in vivo biomarker of mitochondrial membrane potential (ΔΨ) and oxidative phosphorylation (OXPHOS) in lung tumors3. Importantly, by using 18F-BnTP PET imaging we are able to distinguish between OXPHOS dependent and independent lung tumors. Therapeutically, we have demonstrated that 18F-BnTP positive, OXPHOS-dependent LUADs are sensitive to mitochondrial complex I inhibitors. We hypothesize that 18F-BnTP PET imaging can be utilized to functionally profile mitochondrial bioenergetics and adaptive oxidative metabolism in therapy-resistant lung tumors to guide treatment with OXPHOS inhibitors. In aim 1 we will perform an in vivo dissection of mitochondrial bioenergetics in therapy-resistant LUADs. In aim 2 we will perform a structural and functional in vivo analysis of adaptive oxidative metabolism in therapy-resistant KRAS/LKB1 and EGFR mutant LUADs. In Aim 3 we will longitudinally profile oxidative metabolism in LUAD patients with advanced disease. The proposed work has relevance to human health in which we propose that 18F-BnTP PET imaging guided targeting and oxidative metabolism represents a new therapeutic strategy to overcome therapy resistance in patients with KRAS/LKB1 and EGFR mutant tumors.