Dissecting pemetrexed resistance in non-small cell lung carcinoma

剖析非小细胞肺癌培美曲塞耐药性

• 批准号: 10574283
• 负责人: Sumin Kang
• 金额: $ 18.29万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10574283
• 关键词: Antibodies; Attenuated; Biological Assay; Cancer cell line; Cell Line; Cell Proliferation; Cell Survival; Cell surface; Chemoresistance; Chemotherapy-Oncologic Procedure; Clinical; Coupled; DNA biosynthesis; Data; Dihydrofolate Reductase; Dose; Down-Regulation; Enzymes; FGFR1 gene; FGFR3 gene; Failure; Fibroblast Growth Factor Receptors; Folic Acid; Folic Acid Antagonists; Human; In Vitro; Libraries; Link; Lung Adenocarcinoma; MAP Kinase Gene; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Metabolic; Modeling; Molecular; Mus; Non-Small-Cell Lung Carcinoma; Oncogenic; Pathway interactions; Patients; Pemetrexed; Phosphotransferases; Protein Isoforms; Protein Kinase; Protein-Serine-Threonine Kinases; Proteome; RNA interference screen; Regulation; Relapse; Research; Resistance; Resistance development; Ribosomal Protein S6 Kinase; Ribosomes; Role; Scaffolding Protein; Series; Serine; Signal Pathway; Signal Transduction; Therapeutic; Treatment Efficacy; Treatment outcome; Variant; Xenograft procedure; cancer cell; cancer therapy; cancer type; chemotherapy; clinical application; clinical efficacy; combinatorial; folic acid metabolism; improved; in vivo; inhibitor; insight; interest; kinase inhibitor; mouse model; novel; patient derived xenograft model; pharmacologic; response; screening; small hairpin RNA; therapeutic target; therapy outcome; tumor; tumor growth; tumor progression

PROJECT SUMMARY Despite the existence of various therapeutic approaches, chemotherapy is a mainstay of cancer treatment. Pemetrexed-based chemotherapy, a multitargeted antifolate that inhibits folate metabolism, is extensively used to treat non-small cell lung carcinoma (NSCLC), which is the most common type of lung cancer. However, patients often relapse due to the development of resistance, leading to therapeutic failure. Many studies have investigated possible chemoresistance models, yet the precise mechanism is still largely elusive. Oncogenic kinases are well implicated in human cancers and of great clinical interest due to their role in cancer. To better understand the link between kinase-mediated metabolic regulation and pemetrexed resistance, we performed a customized RNAi screen to identify a clinically applicable target kinase that is critical for pemetrexed resistance. We found that inhibition of one of the fibroblast growth factor receptors (FGFR) family, FGFR3, selectively sensitizes NSCLC cells to pemetrexed, leading to decreased cancer cell survival and proliferation. Coupled kinase and metabolic assays revealed that FGFR3 may indirectly activate one of the pemetrexed target enzymes, dihydrofolate reductase (DHFR), in the folate metabolism. Furthermore, global proteome profiling and phospho-signaling array suggested that FGFR3 may be involved in regulating expression or activity of factors in the MAPK pathway including KSR2 and RSK1/2. These suggest that FGFR may provide pemetrexed resistance through modulating folate metabolism and MAPK pathway and is a promising therapeutic target to improve the pemetrexed response. Indeed, pharmacological inhibition of FGFR3 significantly sensitized pemetrexed-resistant NSCLC cell lines to pemetrexed treatment in vitro and in vivo. Our central hypothesis is that FGFR3 confers pemetrexed resistance in NSCLC by regulating the metabolic enzyme DHFR and MAPK pathway. Therefore, FGFR3 inhibitors may represent potent pemetrexed sensitizing agents in NSCLC. Two specific aims are proposed: (1) To decipher the molecular mechanism underlying FGFR3- mediated activation of folate metabolism and MAPK pathway, which confers pemetrexed resistance in NSCLC; (2) To validate FGFR3 signaling as a therapeutic target in treatment of pemetrexed-resistant NSCLC using various NSCLC cell lines and patient-derived xenograft and syngeneic mouse models of lung cancer. This proposal will not only provide information about the role of FGFR3 in pemetrexed resistance but also a new actionable approach to improve the treatment outcome of lung cancer that is not responsive to pemetrexed- based chemotherapy.

项目摘要 尽管存在各种治疗方法，但化疗是癌症治疗的主要手段。 基于培美塞的化疗是一种多靶点抗叶酸剂，可抑制叶酸代谢， 非小细胞肺癌（non-small cell lung carcinoma，NSCLC）是一种常见的肺癌。然而，在这方面， 患者经常由于产生耐药性而复发，导致治疗失败。许多研究 研究了可能的化学抗性模型，但精确的机制在很大程度上仍然难以捉摸。 致癌激酶与人类癌症密切相关，并且由于它们在癌症中的作用而具有极大的临床意义。 癌为了更好地了解激酶介导的代谢调节与培美曲塞耐药之间的联系， 我们进行了一个定制的RNAi筛选，以确定一个临床上适用的靶激酶， 培美曲塞耐药。我们发现，抑制成纤维细胞生长因子受体（FGFR）家族之一， FGFR 3选择性地使NSCLC细胞对培美曲塞敏感，导致癌细胞存活率降低， 增殖偶联激酶和代谢试验显示，FGFR 3可能间接激活其中一种蛋白， 培美曲塞的靶酶是叶酸代谢中的二氢叶酸还原酶（DHFR）。此外，全球 蛋白质组分析和磷酸信号阵列表明FGFR 3可能参与调控表达 或MAPK途径中的因子（包括KSR 2和RSK 1/2）的活性。这表明FGFR可能提供 培美曲塞通过调节叶酸代谢和MAPK途径产生耐药性，是一种有前途的治疗药物 改善培美曲塞反应的目标。事实上，FGFR 3的药理学抑制显著致敏 培美曲塞耐药NSCLC细胞系对培美曲塞治疗的体外和体内研究。 我们的中心假设是，FGFR 3通过调节代谢调节因子， DHFR酶和MAPK途径。因此，FGFR 3抑制剂可能代表培美曲塞的强效致敏作用， NSCLC中的药物。提出了两个具体的目标：（1）破译FGFR 3的分子机制。 介导叶酸代谢和MAPK通路的激活，导致NSCLC对培美曲塞耐药; (2)验证FGFR 3信号传导作为培美塞治疗耐药NSCLC的治疗靶点， 各种NSCLC细胞系和患者来源的肺癌异种移植物和同基因小鼠模型。这 该提案不仅提供了关于FGFR 3在培美曲塞耐药中作用的信息，而且还提供了一种新的 改善培美曲塞无效肺癌治疗结局的可行方法- 基础化疗。