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

摘要（30行）： 这项研究的总体目标是确定有效的基于代谢的诊断和治疗策略 提高非小细胞肺癌（NSCLC）患者的总体生存率。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并开发 针对治疗耐药的氧化线粒体代谢的基于代谢的治疗策略 KRAS/LKB1 和 EGFR 突变肺肿瘤。 2021 年，非小细胞肺癌将夺走美国约 130,000 人的生命。 肺癌 肿瘤通常具有高突变负担，通常需要针对致癌基因的单一药物治疗 驱动突变不成功。此外，肺腺癌（LUAD）的代谢活跃亚型 KRAS 和 LKB1 或 EGFR 突变常常对免疫治疗方法产生耐药性。然而， 无论检查点抑制剂或靶向治疗的最初益处如何，大多数患者都会 最终产生对治疗的抵抗力。我们合理化了克服治疗耐药性的不同方法 非小细胞肺癌（NSCLC）——即根据肿瘤的代谢特征对肿瘤进行分类。在这里，肿瘤被分组并 其目标是通过代谢依赖性而不仅仅是通过基因改变。 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和氧化磷酸化 (OXPHOS) 的场所，并且是必不可少的 用于肺部肿瘤的发生和维持。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了解非小细胞肺癌中线粒体生物能量的调节方式。致地址 为了弥补这一差距，我们利用 PET 成像示踪剂 18F-BnTP 对肺肿瘤中的线粒体活性进行了功能成像 并证明它是线粒体膜电位 (ΔΨ) 和氧化的体内生物标志物 肺部肿瘤中的磷酸化（OXPHOS）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 成像可用于功能性分析线粒体生物能学， 治疗耐药性肺部肿瘤的适应性氧化代谢，指导 OXPHOS 抑制剂的治疗。在 目标 1 我们将对治疗耐药 LUAD 中的线粒体生物能进行体内解剖。目标2 我们将对治疗耐药的适应性氧化代谢进行结构和功能体内分析 KRAS/LKB1 和 EGFR 突变 LUAD。在目标 3 中，我们将纵向分析 LUAD 中的氧化代谢 患有晚期疾病的患者。拟议的工作与人类健康相关，我们建议 18F-BnTP PET 成像引导靶向和氧化代谢代表了一种新的治疗策略 克服 KRAS/LKB1 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.