Project 1: Targeting autophagy for the treatment of KRAS-mutant PDAC

项目1：靶向自噬治疗KRAS突变型PDAC

• 批准号: 10705570
• 负责人: CHANNING J. DER
• 金额: $ 41.52万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705570
• 关键词: Acute; Aftercare; Automobile Driving; Autophagocytosis; BRAF gene; Biological Markers; Biopsy; CRISPR/Cas technology; Caring; Cells; Chronic; Clinical; Clinical Trials; Combined Modality Therapy; Cytostatics; Development; Eating; Evaluation; Future; Gene Expression; Genes; Genetic; Genome; Genomic Library; Glycolysis; Goals; Growth; Growth Inhibitors; Hydroxychloroquine; Immune; Immune response; Impairment; KRAS2 gene; MEK inhibition; MEKs; Malignant Neoplasms; Malignant neoplasm of pancreas; Metabolic; Metabolism; Mitochondria; Mitogen-Activated Protein Kinases; Modeling; Mus; Mutation; Normal tissue morphology; Nutrient; Oncology; Organelles; Organoids; Paclitaxel; Pancreas; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Pharmaceutical Preparations; Phase II Clinical Trials; Phenocopy; Pre-Clinical Model; Process; Protein Kinase; Publishing; Randomized; Recycling; Resistance; Signal Transduction; Source; Toxic effect; Up-Regulation; advanced pancreatic cancer; anti-tumor immune response; arm; cancer cell; clinical efficacy; cytokine; cytotoxic; cytotoxicity; effective therapy; effectiveness evaluation; experimental study; gemcitabine; genetic profiling; improved; inhibition of autophagy; inhibitor; innovation; loss of function; macromolecule; mouse model; mutant; novel; pancreatic cancer model; pancreatic cancer patients; pancreatic ductal adenocarcinoma model; pharmacologic; phase 2 study; research clinical testing; resistance gene; resistance mechanism; response; response biomarker; small molecule libraries; standard of care; targeted treatment; therapeutic target; therapeutically effective; tumor; tumorigenic

PROJECT 1: ABSTRACT Autophagy is a self-degradation process whereby cancer cells recycle defective organelles and macromolecules as a nutrient source to support their increased metabolic needs. Autophagy is elevated and essential for the tumorigenic growth of KRAS-mutant pancreatic ductal adenocarcinoma (PDAC), providing the rationale for clinical evaluation of the autophagy inhibitor hydroxychloroquine (HCQ) for PDAC. Disappointingly, when used as monotherapy in combination with standard of care, HCQ has shown limited to no clinical efficacy for PDAC. We recently determined that the treatment of PDAC with inhibitors of the key KRAS effector pathway, the RAF- MEK-ERK mitogenic activated protein kinase cascade, unexpectedly caused further elevation of autophagy, rendering PDAC acutely dependent on autophagy, and hypersensitive to autophagy inhibition. We determined that ERK inhibition impaired other critical metabolic processes (glycolysis, mitochondrial function) that then led to compensatory upregulation of autophagy. Our findings, together with essentially identical conclusions by another independent co-published study, has led to our initiation of two clinical trials evaluating two approved MEK inhibitors (trametinib, binimetinib) in combination with HCQ for PDAC. Since our recent studies suggest that ERK inhibitors will have superior activity in PDAC, this has prompted our initiation of a phase II clinical trial with the ERK inhibitor LY3214996 in combination with HCQ for metastatic PDAC (Aim 1). While early observations from compassionate care utilization of this combination support a significant clinical impact for this combination, our preliminary studies (Aims 2 and 3) support our premise that we can improve upon this therapy. Aim 2 studies are based on our application of a 2,500-gene druggable genome CRISPR-Cas9 genetic-loss-of- function screen to identify genes that modulate HCQ anti-tumor activity. The identified hits that either enhance or reduce HCQ growth inhibitor activity represent candidate combinations or biomarkers for HCQ resistance, respectively. The biomarkers for response can then be applied to tumor biopsies collected in the Aim 1 clinical trial evaluation. Aim 3 studies involve our application of a chemical library screen using a 525-oncology drug set to identify combinations that enhance the cytotoxicity of HCQ. Together, combinations that arise from Aims 2 and 3 studies will then be advanced to Aim 4 studies, where we will apply PDAC organoid or orthotopic mouse tumor models to identify combinations for future clinical evaluation. Since we have found that ERK MAPK inhibition causes tumor-associated gene expression changes that can lead to an improved anti-tumor immune response, we will also evaluate the impact of our combinations on tumor cytokine expression and on tumor- associated immune cells. In summary, our studies will develop novel combination therapies to target autophagy for the treatment of KRAS-mutant PDAC.

项目1：摘要 自噬是一种自我降解过程，癌细胞借此回收有缺陷的细胞器和大分子 作为一种营养来源，以支持他们增加的代谢需求。自噬是升高的，并且对于 KRAS突变型胰腺导管腺癌（PDAC）的致瘤性生长，提供了 自噬抑制剂羟氯喹（HCQ）用于PDAC的临床评价。令人不安的是，当使用 HCQ作为单一疗法与标准治疗的组合，对PDAC显示出有限的临床功效或没有临床功效。 我们最近确定，用关键的KRAS效应子途径RAF-1抑制剂治疗PDAC是有效的。 MEK-ERK促有丝分裂活化蛋白激酶级联，意外地引起自噬的进一步升高， 使得PDAC严重依赖于自噬，并且对自噬抑制高度敏感。我们确定 ERK抑制损害了其他关键的代谢过程（糖酵解，线粒体功能）， 自噬的补偿性上调。我们的发现，以及基本相同的结论， 另一项独立的共同发表的研究，导致我们启动了两项临床试验，评估两种批准的 MEK抑制剂（曲美替尼、比尼替尼）联合HCQ治疗PDAC。因为我们最近的研究表明 ERK抑制剂在PDAC中具有上级活性，这促使我们启动了II期临床试验 ERK抑制剂LY 3214996与HCQ组合用于转移性PDAC（目的1）。虽然早期 对该组合的同情护理使用的观察支持了对该组合的显著临床影响。 结合，我们的初步研究（目的2和3）支持我们的前提，我们可以改善这种疗法。 目的2研究是基于我们应用2,500个基因的可药物化基因组CRISPR-Cas9基因缺失， 功能筛选以鉴定调节HCQ抗肿瘤活性的基因。所识别的命中， 或降低HCQ生长抑制剂活性代表HCQ抗性的候选组合或生物标志物， 分别然后可以将用于响应的生物标志物应用于在Aim 1临床试验中收集的肿瘤活检。 试验评价Aim 3研究涉及我们使用525肿瘤药物集进行化学库筛选的应用 以鉴定增强HCQ细胞毒性的组合。总之，目标2所产生的组合 3项研究将进入Aim 4研究，我们将应用PDAC类器官或原位小鼠 肿瘤模型，以确定未来临床评估的组合。由于我们已经发现ERK MAPK 抑制引起肿瘤相关基因表达变化，可导致抗肿瘤免疫改善 我们还将评估我们的组合对肿瘤细胞因子表达和肿瘤细胞增殖的影响。 相关的免疫细胞。总之，我们的研究将开发针对自噬的新型联合疗法 治疗KRAS突变型PDAC