The Role of HER3 and IGF1R, through Non-Classical Ras Signaling, on Malignant Peripheral Nerve Sheath Tumor Progression

HER3 和 IGF1R 通过非经典 Ras 信号传导对恶性周围神经鞘肿瘤进展的作用

基本信息

批准号：
9908563
负责人：
Shannon Weber Doutt
金额：
$ 4.77万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9908563
关键词：
Ablation Affect Automobile Driving Biological Process Canertinib Cell Lineage Cell Survival Cells Clustered Regularly Interspaced Short Palindromic Repeats Common Neoplasm Coupled Data Disease Dominant-Negative Mutation ERBB3 gene Event Excision Family Family member Feedback Financial compensation Genetic Screening Growth Image Immunodeficient Mouse In Vitro Individual Insulin-Like-Growth Factor I Receptor KRAS2 gene Knock-out Laboratories Link Luciferases MEKs Malignant Neoplasms Migration Assay NF1 gene Neoplasm Metastasis Nerve Neurofibromatosis 1 Neurofibrosarcoma Operative Surgical Procedures PI3K/AKT Pathway interactions Patients Pharmacology Phenotype Phosphotransferases Process Protein Isoforms Proteins R-ras protein Radiation therapy Receptor Protein-Tyrosine Kinases Recurrence Role Schwann Cells Signal Pathway Signal Transduction Signal Transduction Pathway Soft tissue sarcoma Spindle Cell Neoplasm Testing Therapeutic Tumor Cell Invasion Tumor Cell Line Validation Western Blotting Xenograft procedure chemotherapy combinatorial drug sensitivity effective therapy established cell line experimental study expression vector imaging system in vivo inhibitor/antagonist live cell imaging migration neoplastic cell novel novel strategies outcome forecast ras Proteins receptor receptor expression sarcoma targeted treatment therapeutic target three dimensional cell culture tumor tumor growth tumor progression tumor xenograft

项目摘要

PROJECT SUMMARY/ABSTRACT Currently, no effective treatment for Malignant Peripheral Nerve Sheath Tumors (MPNSTs) exists outside of radical resection, which has poor long-term survival, putting patients with this aggressive spindle cell neoplasm in dire need of an effective treatment. Loss of Neurofibromin 1 (NF1) and subsequent Ras activation are hallmarks of this disease. Proposed therapies targeting Ras and its downstream effectors have failed, so we are exploring targeting upstream activators of Ras, specifically Receptor Tyrosine Kinases (RTKs), which are activated in a positive feedback loop as a result of NF1 loss. Using pharmacologic and genetic screens, we assessed the contribution of all 58 RTKs in MPNST proliferation and survival, identifying the HER receptor HER3 and Insulin-like Growth Factor 1 Receptor (IGF1R) as critical for MPNST progression. Subsequent validation experiments with the broad spectrum HER inhibitor canertinib and an IGF1R inhibitor, picropodophylin (PPP), showed a decrease in proliferation of MPNST cells, with a statistically significant decrease in cell viability with combination treatment. Levels of activated Ras and phosphorylated Erk1/2 were also reduced following combinatorial treatment. While our preliminary data show a decrease in proliferation of MPNST cells and reduced expression of IGF1R initially after HER3 ablation, we observe that in late passage cells, there is a subsequent increase in IGF1R expression and a rescue in proliferation, suggesting compensation and cooperation between the two RTKs. Our preliminary studies also link HER3 to R-Ras family member proteins, specifically linking loss of HER3 with reduced activation of activated R-Ras and R-Ras2. We have explored the role of R-Ras in MPNST migration and invasion, finding a decrease in migration and loss of invasive phenotype after loss of R-Ras family members (via dominant negative vector expression). We propose to rigorously test the hypothesis that HER3 and IGF1R cooperatively promote MPNST progression and modulate migration and invasion through isoform-specific activation of R-Ras proteins and downstream signal transduction pathways of Raf/MEK/ERK and PI3K/AKT; this will be accomplished through 2 aims: first, examining the cooperation of HER3 and IGF1R as activators of Ras isoforms to drive proliferation, migration and invasion, and secondly by testing combinatorial therapy with HER3 and IGF1R inhibitors as a novel approach to effectively inhibit tumor growth when compared to monotherapy with either agent alone. We hope to elucidate the mechanism of RTKs and Ras isoforms on MPNST progression, coupled with identifying a novel combinatorial therapeutic strategy for those suffering from this and other NF1 driven diseases.

项目概要/摘要目前，除了恶性周围神经鞘瘤（MPNST）外，尚无有效的治疗方法。根治性切除术的长期生存率很差，患有这种侵袭性梭形细胞的患者会产生肿瘤神经纤维蛋白 1 (NF1) 缺失和随后的 Ras 激活迫切需要有效的治疗。针对 Ras 及其下游效应子的拟议疗法失败了，因此我们正在探索 Ras 的上游激活剂，特别是受体酪氨酸激酶 (RTK)，它们是由于 NF1 缺失，我们使用药理学和遗传筛选激活了正反馈循环。评估了所有 58 个 RTK 对 MPNST 增殖和存活的贡献，确定了 HER 受体 HER3 和胰岛素样生长因子 1 受体 (IGF1R) 对于 MPNST 的后续进展至关重要。使用广谱 HER 抑制剂 canertinib 和 IGF1R 抑制剂进行验证实验，鬼臼苦素 (PPP) 显示 MPNST 细胞增殖减少，具有统计学显着性联合治疗后细胞活力下降。组合治疗后也减少了，而我们的初步数据显示增殖减少。 MPNST 细胞和 HER3 消融后最初的 IGF1R 表达降低，我们观察到在晚期传代细胞中，IGF1R 表达随后增加，增殖得到挽救，这表明我们的初步研究也将 HER3 与 R-Ras 家族联系起来。成员蛋白，特别将 HER3 的丢失与活化的 R-Ras 和 R-Ras2 的激活减少联系起来。研究人员探索了 R-Ras 在 MPNST 迁移和侵袭中的作用，发现迁移和丢失的减少 R-Ras 家族成员丢失后的侵袭表型（通过显性失活载体表达）。提出严格检验HER3和IGF1R协同促进MPNST的假设 R-Ras Raf/MEK/ERK 和 PI3K/AKT 的蛋白质和下游信号转导通路；通过两个目标实现：首先，检查 HER3 和 IGF1R 作为 Ras 激活剂的合作亚型来驱动增殖、迁移和侵袭，其次通过测试与与其他药物相比，HER3 和 IGF1R 抑制剂是一种有效抑制肿瘤生长的新方法我们希望阐明 RTK 和 Ras 亚型的单药治疗机制。 MPNST 进展，并为患者确定一种新的组合治疗策略来自这种疾病和其他 NF1 驱动的疾病。