Effects of tumor-microbial interactions on EGFR-targeted therapy resistance in lung cancer

肿瘤-微生物相互作用对肺癌EGFR靶向治疗耐药的影响

• 批准号: 10448358
• 负责人: DAN J RAZ
• 金额: $ 20.16万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-09 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10448358
• 关键词: Affect; Bacteria; Biological Assay; Cancer Patient; Cancer cell line; Cell Proliferation; Cells; Chryseobacterium; Collaborations; Drug Efflux; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Generations; Genotype; Human; Laboratories; Lead; Light; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Molecular; Mutate; Mutation; Pathway interactions; Patients; Proteins; Research; Research Design; Resistance; Scientist; Surgeon; Techniques; Testing; actionable mutation; cancer therapy; cell growth; effective therapy; experimental study; fast protein liquid chromatography; inhibitor therapy; innovation; liquid chromatography mass spectrometry; lung cancer cell; microorganism interaction; new therapeutic target; novel strategies; novel therapeutics; pain reduction; preconditioning; predicting response; prevent; resistance mechanism; standard of care; targeted treatment; therapy development; therapy resistant; tumor; tumor microbiome

PROJECT SUMMARY/ABSTRACT We are in desperate need of novel therapies for treating lung cancer, particularly ones that can overcome treatment resistance. Intra-tumoral bacteria are emerging as a cause of therapy resistance in cancer. We recently discovered that a specific bacteria, Chryseobacterium indologenes, identified within human lung cancers, leads to resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI). The third-generation EGFR-TKI osimertinib is the standard of care for first-line treatment of patients with advanced EGFR-mutated lung cancer. Mechanisms of resistance to osimertinib are poorly understood. In this proposal we seek to understand the mechanisms by which intra-tumoral Chryseobacterium (ITC) leads to resistance to osimertinib. The long-term objective of this proposal is to develop new therapies targeting the effects of ITC to prevent resistance to EGFR-TKIs. The objective of this proposal is to identify the mechanism responsible for the effect of ITC on sensitivity to osimertinib, and to determine if other species of Chryseobacterium also lead to EGFR-TKI resistance. Our central hypothesis is that ITC leads to resistance to osimertinib via secreted proteins in some patients with EGFR-mutated lung cancer. In Aim 1, we will validate the candidate proteins that we previously identified in Chryseobacterium preconditioned medium (C-PCM) as potentially responsible for osimertinib resistance using fast protein liquid chromatography and mass spectrometry, by testing the purified candidate proteins in EGFR-mutated lung cancer cell lines in the presence/absence of osimertinib. Next, we will determine the mechanism by which ITC affects resistance to EGFR-TKIs, by investigating the effects of C-PCM on (1) drug efflux, (2) re-activation of the EGFR pathway or (3) activation of molecular pathways other than EGFR. These experiments will help us understand how Chryseobacterium indologenes leads to osimertinib, allowing us to eventually develop new therapies to prevent or overcome osimertinib resistance. In Aim 2, we will determine if Chryseobacterium species other than indologenes lead to EGFR-TKI resistance in PC9 cells. We will obtain commercially available Chryseobacterium meningoseptica, angstadti, gleum, indoltheticum, scophthalmum, diehli, shigense, and balustinum species which will be cultured and genotyped using PCR. Preconditioned medium (PCM) from each species will then be serially diluted and added to PC9 cells with and without osimertinib, and effects on cell growth will be determined using cell proliferation assays. These experiments will help us understand which other Chryseobacterium species lead to osimertinib and may shed additional light on mechanisms of osimertinib resistance. The proposed research is potentially transformative as it may lead to the discovery of a new approach to overcoming resistance to EGFR-TKIs in patients with EGFR- mutated lung cancer through targeting specific tumor-microbial interactions that lead to treatment resistance.

项目总结/摘要 我们迫切需要治疗肺癌的新疗法，特别是那些可以克服 治疗阻力肿瘤内细菌正在成为癌症治疗耐药性的一个原因。我们 最近发现，一种特殊的细菌，产吲哚金黄杆菌，在人类肺部发现， 癌症，导致对表皮生长因子受体（EGFR）酪氨酸激酶抑制剂（TKI）的抗性。的 第三代EGFR-TKI奥希替尼是晚期乳腺癌患者一线治疗的标准治疗， EGFR突变型肺癌对奥希替尼耐药的机制知之甚少。在本提案中，我们 寻求了解肿瘤内金黄杆菌（ITC）导致对 奥希替尼。该提案的长期目标是开发针对ITC影响的新疗法， 防止对EGFR-TKI的耐药性。本建议的目的是确定负责以下事项的机制： ITC对奥希替尼敏感性的影响，并确定其他金黄杆菌属物种是否也导致 EGFR-TKI耐药。我们的中心假设是ITC通过分泌蛋白导致对奥希替尼的耐药性 EGFR突变型肺癌患者的癌症。在目标1中，我们将验证候选蛋白质， 先前在金黄杆菌预处理培养基（C-PCM）中鉴定为可能导致 使用快速蛋白质液相色谱和质谱法，通过检测纯化的 在存在/不存在奥希替尼的情况下EGFR突变的肺癌细胞系中的候选蛋白。接下来我们就 通过研究C-PCM的作用，确定ITC影响EGFR-TKI耐药性的机制 （1）药物外排，（2）EGFR途径的再激活或（3）除 EGFR对这些实验将帮助我们了解产吲哚金杆菌如何导致奥希替尼， 使我们最终能够开发新的疗法来预防或克服奥希替尼耐药性。在目标2中，我们将 确定除吲哚基因外的金黄杆菌属是否导致PC 9细胞中的EGFR-TKI抗性。我们 将获得可商购的脑膜炎黄杆菌、安氏黄杆菌、革兰氏黄杆菌、吲哚黄杆菌， scophthalmum、diehli、shigense和balustinum物种，将培养这些物种并使用PCR进行基因分型。 然后将来自每个物种的预处理培养基（PCM）连续稀释，并加入到PC 9细胞中， 而不使用奥希替尼，并且将使用细胞增殖测定来确定对细胞生长的影响。这些 实验将帮助我们了解哪些其他金黄杆菌属物种导致奥希替尼， 关于奥希替尼耐药机制的更多信息。拟议的研究具有潜在的变革性， 这可能会导致发现一种新的方法来克服EGFR患者对EGFR-TKI的耐药性， 通过靶向特定的肿瘤-微生物相互作用，导致治疗耐药性，来治疗突变的肺癌。