NF2-associated meningiomas: From omics discovery to targeted therapy

NF2 相关脑膜瘤：从组学发现到靶向治疗

基本信息

批准号：
10249966
负责人：
VIJAYA RAMESH
金额：
$ 63.28万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
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：
2020-09-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10249966
关键词：
14q APLN gene Acoustic Neuroma Adult Alleles Anaplastic Meningioma Animal Model Arachnoid mater Back Benign Meningiomas Biological CRISPR/Cas technology Cell Line Cell model Cells Clinical Clinical Trials Collaborations Dasatinib Data Drug Combinations Drug Screening Drug Targeting ERBB3 gene Enzymes Eph Family Receptors Exhibits FRAP1 gene Future Genetic Heterogeneity Genetic Transcription Genomic Instability Growth Growth Factor Heregulin Human IGF1R gene Implant In Vitro Intracranial Neoplasms Ligands Luciferases Maximum Tolerated Dose Measures Modeling Monitor Morbidity - disease rate Mus Mutation NRG1 gene Names Neuregulins Neurofibromatosis 2 Null Lymphocytes Ohio Operative Surgical Procedures Outcome Study Pathway interactions Pediatric Hospitals Pharmaceutical Preparations Pharmacologic Substance Pharmacotherapy Phosphotransferases Play Proteasome Inhibitor Protein Tyrosine Kinase Proto-Oncogene Protein c-kit Public Health Publishing Radiation Randomized Receptor Protein-Tyrosine Kinases Regulation Research Resected Role SDZ RAD Screening Result Signal Pathway Signal Transduction Skull Base Neoplasms System Systemic Therapy Testing Time Translating Translational Research Tumor Suppressor Genes Ubiquitin Ubiquitin-Activating Enzymes United States National Institutes of Health Up-Regulation Work antitumor effect base bioluminescence imaging chromosome 22q loss clinical care clinical development comparative genomic hybridization cytotoxic drug efficacy effective therapy follow-up genome editing heparin-binding EGF-like growth factor in vivo in vivo Model in vivo evaluation inhibitor/antagonist mTOR Inhibitor member meningioma mortality mouse model multicatalytic endopeptidase complex neoplastic cell novel novel therapeutic intervention pre-clinical response single-cell RNA sequencing small molecule inhibitor src-Family Kinases targeted treatment therapeutic target transcriptome transcriptome sequencing tumor tumor growth tumor heterogeneity

项目摘要

Neurofibromatosis 2 (NF2) is characterized by vestibular schwannomas, and meningiomas that show loss of the NF2 tumor suppressor gene. Meningiomas arise from meningothelial arachnoid cap cells and are the most common primary intracranial tumors in adults. NF2 inactivation is frequently associated with sporadic meningiomas, particularly in primary atypical (WHO grade II) tumors. Meningiomas that progress despite surgery and radiation are responsible for significant morbidity and mortality. Therefore, effective systemic therapies are greatly needed. For meningioma modeling and preclinical drug screening, we employ human primary meningioma (MN) cell lines derived from surgically resected tumors and CRISPR-Cas9 genome-edited isogenic, human arachnoidal cell lines (ACs) that either express or lack NF2. Employing these models, we established that NF2-deficient meningiomas reveal aberrant activation of mTORC1/mTORC2 signaling, which led to clinical trials for NF2 and meningiomas. Recently, we undertook large-scale kinome, transcriptome and drug screening studies in our AC and MN cell models to identify other potential targets. Kinome and transcriptome data revealed increased activation and expression of several EPH receptor family tyrosine kinases (EPH-RTKs), Src family kinase members (SFKs) and c-KIT, which are all targets of dasatinib, which is recently published. Our ‘omics approach also identified other interesting candidates, including ligands NRG1, HBEGF and apelin, and AMPK-related kinases, particularly NUAK2, to be consistently upregulated in the kinome and transcriptome of NF2-null AC/MN cells. Our recent results suggest that NRG1 expression and ERBB3 signaling is regulated by mTORC1 signaling. We plan to examine whether NRG1 alone or factors such as HBEGF and APLN are also regulated by mTOR or play a role in downstream signaling in NF2-null MN cells. We propose to understand the mechanism and biological significance of elevated expression and activation of NUAK2 in meningioma. Further, our large-scale drug screening efforts, in collaboration with NIH-NCATS, revealed a set of proteasome pathway related drugs exhibiting cytotoxic effects in NF2-null cells. Here we propose to examine three different drugs targeting the proteasome (provided by Millennium-Takeda Pharmaceuticals), alone and combined with TAK-228 (mTOR inhibitor) in 5 grade I, 5 grade II and 5 grade III MN lines with NF2 loss. More importantly, we propose to undertake single cell RNAseq along with array CGH in NF2-deficient meningiomas and their corresponding primary cell lines to define tumor heterogeneity and correlate with drug response. Anti-tumor efficacy of proteasome drugs will also be evaluated in orthotopic NF2- deficient benign and malignant meningioma models. Our overall approach of (i) leveraging the ‘omics and drug screening results, (ii) characterizing tumor heterogeneity and correlating with drug response, and (iii) testing potential drugs in orthotopic NF2-deficient in vivo models is timely, novel and will provide a framework to pursue new avenues in NF2 and meningioma research for clinical care.

神经纤维瘤病 2 (NF2) 的特征是前庭神经鞘瘤和脑膜瘤，表现为神经纤维瘤丧失 NF2肿瘤抑制基因脑膜瘤起源于脑膜上皮蛛网膜帽细胞，是最常见的。成人常见的原发性颅内肿瘤通常与散发性相关。脑膜瘤，特别是原发性非典型（WHO II 级）肿瘤，尽管如此，仍会进展。手术和放疗导致显着的发病率和死亡率，因此，有效的全身治疗。对于脑膜瘤建模和临床前药物筛选，我们非常需要治疗方法。原发性脑膜瘤 (MN) 细胞系源自手术切除的肿瘤并经过 CRISPR-Cas9 基因组编辑我们采用表达或缺乏 NF2 的同基因人类蛛网膜细胞系 (AC)。确定 NF2 缺陷的脑膜瘤揭示 mTORC1/mTORC2 信号传导的异常激活，这最近，我们进行了大规模的激酶组、转录组和脑膜瘤临床试验。在我们的 AC 和 MN 细胞模型中进行药物筛选研究，以确定其他潜在的靶标。转录组数据显示几个 EPH 受体家族酪氨酸的激活和表达增加激酶 (EPH-RTK)、Src 家族激酶成员 (SFK) 和 c-KIT，这些都是达沙替尼的靶点，我们最近发表的“组学方法”还发现了其他有趣的候选者，包括配体 NRG1， HBEGF 和 apelin 以及 AMPK 相关激酶，特别是 NUAK2，在我们最近的结果表明，NRG1 表达和 NF2 缺失 AC/MN 细胞的激酶组和转录组有关。 ERBB3 信号传导受 mTORC1 信号传导调节，我们计划检查 NRG1 是否单独调节或受其他因素调节。因为 HBEGF 和 APLN 也受 mTOR 调节或在 NF2 缺失 MN 细胞的下游信号传导中发挥作用。我们建议了解表达升高和激活的机制和生物学意义此外，我们与 NIH-NCATS 合作进行了大规模的药物筛选工作，在此，我们揭示了一组在 NF2 缺失细胞中表现出细胞毒性作用的蛋白酶体途径相关药物。提议检查三种不同的靶向蛋白酶体的药物（由 Millennium-Takeda 提供）药物），单独使用以及与 TAK-228（mTOR 抑制剂）联用，5 个 I 级、5 个 II 级和 5 个 III 级具有 NF2 丢失的 MN 系更重要的是，我们建议进行单细胞 RNAseq 和阵列 CGH。在 NF2 缺陷型脑膜瘤及其相应的原代细胞系中，以确定肿瘤异质性和蛋白酶体药物的抗肿瘤功效也将在原位 NF2- 中进行评估。我们的总体方法是（i）利用“组学和药物”。筛选结果，(ii) 表征肿瘤异质性并与药物反应相关，以及 (iii) 测试原位 NF2 缺陷体内模型中的潜在药物是及时的、新颖的，并将提供一个框架寻求 NF2 和脑膜瘤研究临床护理的新途径。