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.

抽象的 胰腺导管腺癌（PDAC）对治疗特别有抵抗力，通常呈现为 转移性疾病。以缺氧，致密的基质和代谢重新布线，原始方法和 迫切需要组合策略。我们建议研究抑制氧化还原信号蛋白 以及通过撞击肿瘤使用的临界途径来选择性地杀死肿瘤的药物组合 存活。氧化还原因子1（Ref-1）调节驱动胰腺癌的各种转录因子的活性 细胞增殖和耐药性以及与细胞代谢有关的基因。在缺氧下，抑制作用 Ref-1显着扰动的代谢途径（TCA循环和OXPHOS）以及HIF调节的基因，以及 因此，胰腺癌共培养球体和异种移植物的生长减慢了。第一代ref-1 抑制剂（APX3330）完成了I期试验，并证明了32％的响应，预测PK和目标 参与没有明显的毒性。六名治疗患者有疾病稳定 长时间（> 250天）。基于令人鼓舞的I阶段数据和详细的结构活动 关系（SAR）计划，我们还确定了处于潜在客户优化的下一代Ref-1抑制剂 阶段，一种筛选对Ref-1抑制敏感的患者的策略，以及分子靶标的 可能与Ref-1抑制协同作用。但是，抗性的自适应机制最终出现 有针对性的治疗，因此我们还将重点关注新型组合的发展。我们的假设是 针对Ref-1的氧化还原功能，并且在机械设计的组合疗法中会诱导 代谢致死性和抑制胰腺癌的生长和转移。在AIM 1中，代谢的识别 与Ref-1抑制结果相关的癌细胞/组织的特征 代谢靶标提高了Ref-1抑制的疗效。我们最近开发的计算预测指标 细胞的代谢通量将用于研究由于PDAC细胞中REF-1抑制作用而导致的代谢变化 单细胞水平。在AIM 2中，NMR建立了Ref-1和新类似物，功效，毒性，毒性的直接相互作用 和代谢稳定性研究将使我们能够促进候选人体内研究的顶级铅候选者 选择（NIH里程碑4）和IND（研究新药）提交导致最终I期试验。 最后，在AIM 3中，将使用Ref-1评估临床前组合疗法中的REF-1来克服适应性 反抗。进一步预测可能会扰动与Ref-1抑制协同作用的代谢节点 将使用AIM1中描述的细胞代谢通量的代谢致死性，计算预测指标。这 Ref-1单独和新组合的功效将在体外和小鼠试验中研究 在体内设计。总而言之，对于杀死PDAC的精确方法，我们将提供有效而有选择性的Ref-1 抑制剂并结合新型的代谢生物信息学和药物组合，以增强功效 对现场和临床治疗学的重大影响。