Role of Receptor Tyrosine Kinase cross-talk in colorectal cancer

受体酪氨酸激酶串扰在结直肠癌中的作用

• 批准号: 10404504
• 负责人: Bhuminder Singh
• 金额: $ 46.38万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-12 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10404504
• 关键词: 3-Dimensional; Address; Antibodies; BRAF gene; Biological Markers; Cancer Etiology; Cancer Model; Cessation of life; Cetuximab; Chemicals; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collagen Type I; Colon; Colorectal Cancer; Drug Combinations; ERBB3 gene; Effectiveness; Epidermal Growth Factor Receptor; Exhibits; Gene Expression Profile; Genetic; Grant; High Prevalence; Human; Hypersensitivity; Immunocompetent; In Vitro; Individual; KRAS2 gene; Knock-out; Ligands; Link; MAP Kinase Gene; Mediating; Modification; Monoclonal Antibodies; Morphology; Mus; Mutation; NRG1 gene; Neoplasm Metastasis; Nude Mice; Oncogenic; Operative Surgical Procedures; Organoids; Patient Selection; Patients; Pharmacology; Pharmacotherapy; Phase I/II Clinical Trial; Phenotype; Phosphorylation; Play; Positron-Emission Tomography; Prevalence; Prognosis; Progression-Free Survivals; Proteins; Ras/Raf; Reaction; Receptor Protein-Tyrosine Kinases; Research; Research Design; Research Personnel; Resistance; Resistance profile; Role; Sampling; Signal Pathway; Signal Transduction; Southeastern United States; System; Testing; Therapeutic Monoclonal Antibodies; Therapeutic antibodies; Tissue Microarray; Tumor Tissue; Tumor-Derived; Tyrosine Kinase Inhibitor; Tyrosine Kinase Receptor Inhibition; Tyrosine Phosphorylation; United States; Unresectable; Work; Xenograft procedure; base; colon cancer cell line; colorectal cancer progression; colorectal cancer treatment; crizotinib; experimental study; imaging probe; improved; in vitro Model; in vivo; in vivo Model; inclusion criteria; inhibitor; neutralizing antibody; non-genetic; novel; overexpression; panitumumab; patient derived xenograft model; patient population; patient stratification; personalized medicine; precision medicine; predicting response; prevent; receptor; receptor upregulation; resistance mechanism; response; small molecule inhibitor; targeted treatment; therapeutic target; three dimensional cell culture; treatment response; treatment strategy; tumor; tumor xenograft; uptake

ABSTRACT: Background: EGF receptor (EGFR) is a receptor tyrosine kinase (RTK) that is overexpressed in over 50% of colorectal cancer (CRC) cases where it is linked to metastasis and poor prognosis. Cetuximab, an EGFR-targeting monoclonal antibody is approved by US-FDA to treat advanced wild-type KRAS CRC. However, cetuximab, as monotherapy, is effective in only about 10% of CRCs and resistance frequently emerges. Thus, there is a pressing need 1) to identify those patients most likely to respond (or not respond) to cetuximab and 2) to devise treatment strategies that would prevent resistance and/or enhance cetuximab response. We propose to address these needs by aggressively pursuing our observations that enhanced activity of multiple RTKs (e.g. MET and RON) confers de novo and acquired cetuximab resistance, which may be overcome by addition of the dual MET/RON inhibitor, crizotinib. Our central hypothesis is that upregulation of RTK activity confers cetuximab resistance in CRC and that it may be a viable therapeutic target. In Aim 1 we will identify the mechanism of RTK cooperation during CRC progression and cetuximab resistance using genetic and chemical modifications in CRC cell lines and patient- derived xenografts (PDXs) and organoids (PDOs). In Aim 2 we will assess the impact of disrupting RTK cooperation in overcoming resistance to EGFR-directed antibodies. In Aim 3 we will identify rational drug combinations and stratify patient populations to overcome resistance to EGFR- directed mAbs. Study design: We will employ our in vitro 3D culture system for CRC cell lines and PDOs and in vivo nude mice xenografts, PDXs, and immune-competent syngeneic mouse CRC models in this grant. We will also employ complementary approaches (Transwell cultures, tumor tissue microarrays, phospho-RTK arrays, and human CRC samples) in relevant sections. Key proteins (EGFR, MET, RON, ERBB3, HGF, HGFL, NRG1 and other positive and negative RTK and ligand regulators) will be manipulated (overexpression, CRISPR-mediated knockout, ligand stimulation, and chemical and antibody-based inhibition) to characterize individual contribution. These experiments will be recorded at subcellular localization, morphological, and phenotypic levels to tease out key differences. These studies will identify and characterize new modes of cetuximab resistance and its prevalence in CRC, while devising means to prevent or overcoming the resistance. This research will help optimize precision medicine and stratify individuals based on their response/resistance profile for better CRC treatment and management.

摘要： 背景：表皮生长因子受体（EGFR）是一种受体酪氨酸激酶（RTK），在表皮生长因子受体（EGF）中过度表达。 超过50%的结直肠癌（CRC）病例与转移和预后不良有关。 西妥昔单抗是一种靶向EGFR的单克隆抗体，已被美国FDA批准用于治疗晚期 野生型KRAS CRC。然而，西妥昔单抗作为单一疗法，仅在约10%的患者中有效。 CRCs和耐药性经常出现。因此，迫切需要1）确定那些 最有可能对西妥昔单抗有反应（或无反应）的患者; 2）设计治疗策略 这将防止耐药性和/或增强西妥昔单抗反应。我们建议解决这些问题 通过积极追求我们的观察，增强了多个RTK的活动（例如， MET和罗恩）引起新发和获得性西妥昔单抗耐药，可通过以下方法克服： 添加MET/罗恩双重抑制剂克唑替尼。我们的中心假设是， RTK活性赋予西妥昔单抗在CRC中的耐药性，并且它可能是可行的治疗靶点。 在目标1中，我们将确定CRC进展期间RTK合作的机制， 在CRC细胞系和患者中使用遗传和化学修饰的西妥昔单抗抗性 衍生的异种移植物（PDX）和类器官（PDO）。在目标2中，我们将评估破坏 RTK合作克服对EGFR定向抗体的抗性。在目标3中，我们将确定 合理的药物组合和分层的患者人群，以克服耐药EGFR- 定向mAb。 研究设计：我们将使用我们的体外3D培养系统用于CRC细胞系和PDO， 本研究采用体内裸小鼠异种移植物、PDX和具有免疫能力的同基因小鼠CRC模型 格兰特.我们还将采用互补的方法（Transwell培养，肿瘤组织 微阵列、磷酸-RTK阵列和人CRC样品）。关键蛋白质 (EGFR、MET、罗恩、ERBB 3、HGF、HGFL、NRG 1等阳性和阴性RTK及配体 调节子）将被操纵（过表达，CRISPR介导的敲除，配体刺激， 和基于化学和抗体的抑制）来表征个体贡献。这些 实验将在亚细胞定位、形态和表型水平上记录， 找出关键的区别。 这些研究将确定和表征西妥昔单抗耐药的新模式及其 在CRC的患病率，同时设计手段，以防止或克服阻力。本研究 将有助于优化精准医疗，并根据个人的反应/抵抗力对其进行分层 更好的CRC治疗和管理。