In vivo imaging of mitochondria structure and function in therapy resistant lung tumors

摘要（30行）： 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，通常使得靶向致癌基因的单一药剂疗法 驱动突变不成功。此外，肺腺癌（LUAD）的代谢活性亚群 携带KRAS和LKB 1或EGFR突变的患者通常对免疫治疗方法具有抗性。然而，在这方面， 无论最初从检查点抑制剂或靶向治疗中获益，大多数患者将 最终会对治疗产生抗药性我们为克服治疗抵抗的不同方法找理由 在NSCLC中-即通过其代谢特征对肿瘤进行分类。在这里，肿瘤被分组， 通过代谢依赖性而不仅仅是通过基因改变来靶向。NSCLC是一种代谢性 异质性疾病和肿瘤利用糖酵解和氧化线粒体代谢来生长。的 线粒体是细胞生物能量学和氧化磷酸化（OXPHOS）的场所， 肺肿瘤的发生和维持。由于缺乏体内成像探针，我们的知识存在空白 在生理和机制水平上研究NSCLC中线粒体生物能量学是如何调节的。解决 为了弥补这一空白，我们利用PET显像示踪剂18F-BnTP对肺肿瘤中的线粒体活性进行了功能性成像 并证明它是线粒体膜电位（Δ ε）和氧化应激的体内生物标志物。 磷酸化（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18 F-BnTP PET成像可用于功能性分析线粒体生物能量学，并 治疗抗性肺肿瘤的适应性氧化代谢，以指导OXPHOS抑制剂的治疗。在 目的1我们将在体内解剖治疗抗性LUAD中的线粒体生物能量学。在aim 2中 我们将在体内对治疗抵抗的适应性氧化代谢进行结构和功能分析。 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 (30 line): 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.