Metabolic flux analysis and PDX models to understand therapeutic vulnerabilities following inhibition of Ref-1 redox signaling in pancreatic cancer

代谢通量分析和 PDX 模型可了解胰腺癌中 Ref-1 氧化还原信号传导抑制后的治疗脆弱性

• 批准号: 10717281
• 负责人: Melissa L Fishel
• 金额: $ 46.63万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10717281
• 关键词: 3-Dimensional; Adult; Bioinformatics; Cell Line; Cell Proliferation; Cells; Characteristics; Citric Acid Cycle; Clinical; Clinical Trials; Coculture Techniques; Combined Modality Therapy; Critical Pathways; Data; Development; Disease; Dose; Drug Combinations; Drug resistance; Enzymes; Evaluation; Future; Gene Expression; Generations; Genes; Genetic Transcription; Growth; Hypoxia; In Vitro; Individual; Investigational Drugs; Lead; Malignant neoplasm of pancreas; Metabolic; Metabolic Pathway; Metabolic stress; Metabolism; Methods; Mitochondria; Molecular Target; Mus; Neoplasm Metastasis; Organoids; Outcome; Oxidation-Reduction; Pancreatic Ductal Adenocarcinoma; Patients; Pharmaceutical Preparations; Pharmacodynamics; Phase; Phase I Clinical Trials; Play; Proliferating; Reaction; Resistance; Role; Signal Transduction; Signaling Protein; Solid Neoplasm; Testing; Therapeutic; Time; Tissues; Toxic effect; Treatment Efficacy; Triage; United States National Institutes of Health; Xenograft procedure; advanced disease; analog; cancer cell; candidate selection; carbonate dehydratase; clinical development; combinatorial; design; disorder control; improved; in vivo; in vivo Model; inhibitor; knock-down; lead candidate; lead optimization; metabolic abnormality assessment; next generation; novel; novel therapeutics; nutrient deprivation; pancreatic cancer cells; pancreatic cancer patients; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; patient derived xenograft model; patient screening; personalized approach; phase I trial; pre-clinical; preclinical development; programs; resistance mechanism; response; synergism; targeted agent; targeted treatment; therapeutic target; therapy resistant; transcription factor; trial design; tumor; tumor growth; tumor metabolism

ABSTRACT Pancreatic ductal adenocarcinoma (PDAC) is particularly resistant to therapy and typically presents as metastatic disease. Characterized by hypoxia, dense stroma, and metabolic rewiring, original approaches and combination strategies are desperately needed. We propose to investigate inhibition of a redox signaling protein and drug combinations that selectively kill the tumor by impinging on critical pathways the tumor is using to survive. Redox factor-1 (Ref-1) regulates the activity of various transcription factors that drive pancreatic cancer cell proliferation and drug resistance as well as genes involved in cellular metabolism. Under hypoxia, inhibition of Ref-1 significantly perturbed metabolic pathways (TCA cycle and OXPHOS) and HIF-regulated genes, and thus slowed the growth of pancreatic cancer co-culture spheroids and xenografts. The first-generation Ref-1 inhibitor (APX3330) completed phase I trial and demonstrated 32% response, predicted PK, and target engagement with no significant toxicities. There was disease stabilization in six patients with four on treatment for an extended time (>250 days). Based on encouraging phase I data and a detailed structural-activity relationship (SAR) program, we have also identified next generation Ref-1 inhibitors that are at lead optimization stage, a strategy to screen for patients that have sensitivity to Ref-1 inhibition, and molecular targets that are likely to synergize with Ref-1 inhibition. However, adaptive mechanisms of resistance eventually emerge with targeted therapy, therefore we will also focus on the development of novel combinations. Our hypothesis is that targeting the redox function of Ref-1 alone and in mechanistically designed combination therapies will induce metabolic lethality and inhibit pancreatic cancer growth and metastasis. In Aim 1, identification of metabolic characteristics of cancer cells/tissues that associate with the outcome of Ref-1 inhibition and prediction of new metabolic targets to improve the efficacy of Ref-1 inhibition. Our recently developed computational predictor of cell-wise metabolic flux will be used to study the metabolic changes due to Ref-1 inhibition in PDAC cells at the single cell level. In Aim 2, NMR to establish direct interactions of Ref-1 and the new analogues, efficacy, toxicity, and metabolic stability studies will allow us to advance the top lead candidate(s) for in vivo studies for Candidate Selection (NIH Milestone 4) and IND (Investigational New Drug) submission leading to eventual Phase I trial. Lastly in Aim 3, evaluation of Ref-1 in preclinical combination therapy will be used to overcome adaptive resistance. To further predict metabolic nodes that could be perturbed to synergize with Ref-1 inhibition, creating a metabolic lethality, computational predictor of cell-wise metabolic flux described in Aim1 will be used. The efficacy of Ref-1 alone and in new combinations will be investigated using organoids in vitro and the mouse trial design in vivo. In summary, for a precision approach to kill PDAC, we will deliver a potent and selective Ref-1 inhibitor and combine novel metabolic bioinformatics and drug combinations for enhanced efficacy to have a significant impact on the field and clinical therapeutics.

摘要 胰腺导管腺癌(PDAC)对治疗特别耐药，通常表现为 转移性疾病。以缺氧、致密间质和新陈代谢重新连接为特征，原始的入路和 人们迫切需要组合策略。我们建议研究氧化还原信号蛋白的抑制作用。 以及通过冲击肿瘤的关键途径选择性地杀死肿瘤的药物组合 活下来。氧化还原因子-1(Ref-1)调节多种导致胰腺癌的转录因子的活性 细胞增殖和耐药性以及参与细胞代谢的基因。在低氧条件下，抑制作用 REF-1显著干扰代谢途径(TCA周期和OXPHOS)和HIF调节基因，以及 从而减缓了胰腺癌共培养球体和异种移植瘤的生长。第一代参考文献-1 抑制剂(APX3330)完成I期试验，显示32%的有效率、预测的PK和靶向 订婚没有明显的毒性。有6名患者病情稳定，其中4名正在接受治疗。 对于延长的时间(&gt；250天)。基于令人鼓舞的第一阶段数据和详细的结构性活动 关系(SAR)计划，我们还确定了处于领先优化状态的下一代Ref-1抑制剂 阶段，筛选对Ref-1抑制敏感的患者的策略，以及符合以下条件的分子靶点 可能与Ref-1抑制协同作用。然而，适应性的抗性机制最终会随着 靶向治疗，因此我们还将重点开发新的组合。我们的假设是 靶向Ref-1的氧化还原功能单独和在机械设计的联合治疗中将诱导 代谢致命性和抑制胰腺癌的生长和转移。在目标1中，确定代谢 与Ref-1抑制结局相关的癌细胞/组织特征及新基因的预测 代谢靶点，以提高抑制Ref-1的疗效。我们最近开发的计算预报器 细胞水平的代谢流量将用于研究由于抑制Ref-1在PDAC细胞中的代谢变化 单细胞水平。在目标2中，核磁共振建立Ref-1与新类似物的直接相互作用，疗效，毒性， 代谢稳定性研究将使我们能够提拔出领先的候选人(S)进行体内研究 选择(NIH里程碑4)和IND(研究新药)提交导致最终的I期试验。 最后，在目标3中，将使用Ref-1在临床前联合治疗中的评估来克服适应性 抵抗。为了进一步预测可能被干扰以与Ref-1抑制协同的代谢节点，创建 将使用Aim1中描述的代谢致命性、细胞代谢通量的计算预测因子。这个 将在体外和小鼠试验中使用有机化合物来研究Ref-1单独和新组合的疗效 活体设计。总而言之，为了一种精确的方法杀死PDAC，我们将提供一个强大的和选择性的参考-1 抑制剂和结合新的代谢生物信息学和药物组合以增强疗效 对领域和临床治疗产生重大影响。