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

• 批准号: 10316168
• 负责人: Shannon Weber Doutt
• 金额: $ 4.98万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 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/10316168
• 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; Prognosis; 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; live cell imaging; migration; neoplastic cell; novel; novel strategies; ras Proteins; receptor; receptor expression; sarcoma; targeted treatment; therapeutic target; three dimensional cell culture; tumor; tumor growth; tumor progression; tumor xenograft

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.

项目总结/摘要 目前，恶性外周神经鞘瘤（MPNST）的有效治疗方法不存在于 根治性切除术，长期生存率低，使这种侵袭性梭形细胞肿瘤患者 急需有效的治疗神经纤维蛋白1（NF 1）的丢失和随后的Ras激活是 这种疾病的特征。针对Ras及其下游效应物的拟议疗法已经失败，所以我们 正在探索靶向Ras的上游激活剂，特别是受体酪氨酸激酶（RTK）， 由于NF 1的损失，在正反馈回路中被激活。使用药理学和遗传学筛选，我们 评估了所有58种RTK在MPNST增殖和存活中的作用，鉴定了HER受体， HER 3和胰岛素样生长因子1受体（IGF 1 R）对MPNST进展至关重要。后续 使用广谱HER抑制剂canertinib和IGF 1 R抑制剂的验证实验， 苦鬼臼脂素（PPP），显示MPNST细胞增殖的减少，具有统计学显著性差异。 联合处理降低细胞活力。激活的Ras和磷酸化的Erk 1/2水平是 在联合治疗后也降低。虽然我们的初步数据显示， MPNST细胞和IGF 1 R的表达减少，我们观察到在晚期传代中， 细胞中，随后IGF 1 R表达增加，并挽救增殖，表明 两个RTK之间的补偿和合作。我们的初步研究也将HER 3与R-Ras家族联系起来 成员蛋白，特异性地将HER 3的丧失与活化的R-Ras和R-Ras 2的活化降低联系起来。我们 研究了R-Ras在MPNST迁移和侵袭中的作用，发现MPNST的迁移和丢失减少， R-Ras家族成员丢失后的侵袭表型（通过显性负载体表达）。我们 建议严格检验HER 3和IGF 1 R协同促进MPNST的假设 通过R-Ras同种型特异性激活来进行和调节迁移和侵袭 Raf/MEK/ERK和PI 3 K/AKT的蛋白质和下游信号转导途径;这将是 通过2个目标完成：首先，检查HER 3和IGF 1 R作为Ras激活剂的合作 亚型来驱动增殖、迁移和侵袭，并且其次通过测试与 HER 3和IGF 1 R抑制剂作为有效抑制肿瘤生长的新方法， 单独使用任一药剂的单一疗法。我们希望阐明RTKs和Ras异构体在细胞凋亡中的作用机制。 MPNST进展，再加上确定一种新的组合治疗策略，为那些患有 免受这种疾病和其他NF 1驱动的疾病的影响。