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Uncovering treatment targets for peripheral nerve sheath tumor progression in NF1

发现 NF1 周围神经鞘瘤进展的治疗靶点

基本信息

批准号：
10439480
负责人：
DAVID ANDREW LARGAESPADA
金额：
$ 57.74万
依托单位：
UNIVERSITY OF MINNESOTA
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 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10439480
关键词：
Address Benign C57BL/6 Mouse CDKN2A gene Cell Line Cells Chemicals Clustered Regularly Interspaced Short Palindromic Repeats DNA sequencing Detection Development Drug Screening Drug Targeting Environment Epigenetic Process Erinaceidae Evolution Genes Genetic Genetic Diseases Genetic Screening Genetically Engineered Mouse Genotype Goals Heritability Human Image Analysis Immune Immune system Immunocompromised Host In Vitro Lesion Lethal Genes Life Malignant - descriptor Malignant Neoplasms Methods Modeling Modification Molecular Target Morbidity - disease rate Mus Mutation NF1 gene Neoplasms Nerve Nerve Sheath Tumors Neurofibromatoses Neurofibromatosis 1 Neurofibrosarcoma Operative Surgical Procedures Pain Patients Perinatal Peripheral Nerve Sheath Neoplasm Peripheral Nerves Persons Pharmaceutical Preparations Plexiform Neurofibroma Population Pre-Clinical Model Preclinical Testing Premalignant Cell Prevention Process Prognosis Protein Phosphatase 2A Regulatory Subunit PR53 Role Schwann Cells Soft tissue sarcoma Source Syndrome System Testing Therapeutic Uses Tissues Wild Type Mouse base cancer predisposition cell transformation chemotherapy dermal neurofibroma drug candidate exome gene product genetically modified cells human data immunoregulation in vitro Model in vitro testing in vivo in vivo Model indexing induced pluripotent stem cell loss of function mutation molecular targeted therapies mortality mouse model mutant neurofibroma new therapeutic target novel therapeutics prevent recruit sciatic nerve subcutaneous transcriptome transcriptome sequencing transplant model tumor tumor progression tumor-immune system interactions

项目摘要

Project Summary/Abstract Neurofibromatosis type 1 (NF1) syndrome is an autosomal dominant cancer-predisposing syndrome afflicting ~1 in every 3,500 persons worldwide with the majority of patients developing benign plexiform and/or dermal neurofibromas. Plexiform neurofibromas constitute a lifelong source of disfigurement, morbidity and mortality, and have the potential to transform to a malignant peripheral nerve sheath tumor (MPNST), an aggressive soft tissue sarcoma. In fact, approximately 15% of NF1 patients develop poor prognosis MPNSTs, often in the second or third decade of life. Treatment options for MPNSTs are limited to complicated surgical procedures and classical chemotherapy and, so far, molecular targeted therapies have demonstrated limited efficacy. We desperately need new treatment options for the MPNSTs and methods to prevent them from development. It was recently found that plexiform neurofibromas progress to MPNST via an intermediate, “atypical” neurofibroma (ANF) that in 70% of tumors shows heterozygous or homozygous loss of CDKN2A the gene encoding p16INK4a and p14ARF (Beert et al., 2011; Pemov et al., 2018). Our proposal will address critical unmet needs in this field, including better in vitro and in vivo models of ANF and identification of critical vulnerabilities of these cells. To provide a model for preclinical testing and prevention of ANF to MPNST development, we combined Desert hedgehog (Dhh)-Cre driven biallelic deletion of Nf1 with heterozygous loss of Cdkn2a, creating a unique model of transplantable ANF developing within pre-existing neurofibroma (Chaney et al., submitted). We also combined Dhh-Cre driven biallelic deletion of Nf1 and Pten, generating rapidly developing perinatal ANF-like lesions (Keng et al., 2012). ANF from Dhh-Cre;Nf1fl/fl;Cdkn2a+/- mice grafted subcutaneously into immunocompromised hosts grew, after a delay, providing a more rapid, tractable, transformation system. We plan a complete transcriptome and exome analysis in these models (Aim 1a), and further investigate the model by identifying and validating cell populations and markers altered in mouse and human PNF, ANF, and MPNST in unperturbed tissue sections using a new image analysis method called CO-Detection by IndEXing (Aim 1b). Modulation of the immune environment is increasing used therapeutically. We will therefore define the influence of the nerve microenvironment and immune system on progression from ANF to MPNST (Aim 1c). To identify ANF vulnerabilities, we have completed drug and CRISPR-based genetic synthetic lethality screens in isogenic immortalized human Schwann cells that are NF1 wildtype or were made homozygous for NF1 loss of function mutations using gene editing. Candidate drugs that inhibit PP2A, and other novel targets from the drug screening effort, will be tested for their effects in ANF- like cells in vitro (Aim 2a) and, when successful, in our unique GEMMs (Aim 2b). Similarly, our genetic screening effort will be used to define additional vulnerabilities tested in vitro (Aim 2c) and in vivo (Aim 2d).

项目概要/摘要 1 型神经纤维瘤病 (NF1) 综合征是一种常染色体显性遗传癌症诱发综合征全球每 3,500 人中约有 1 人患有良性丛状和/或皮肤病神经纤维瘤。丛状神经纤维瘤是导致毁容、发病和死亡的终生根源，并且有可能转变为恶性周围神经鞘瘤（MPNST），这是一种侵袭性软组织肿瘤组织肉瘤。事实上，大约 15% 的 NF1 患者出现预后不良的 MPNST，通常发生在生命的第二个或第三个十年。 MPNST 的治疗选择仅限于复杂的外科手术传统化疗以及迄今为止的分子靶向治疗已证明疗效有限。我们迫切需要针对 MPNST 的新治疗方案和防止其发展的方法。它最近发现丛状神经纤维瘤通过中间“非典型”进展为 MPNST 神经纤维瘤 (ANF)，70% 的肿瘤显示 CDKN2A 基因杂合或纯合缺失编码 p16INK4a 和 p14ARF（Beert 等人，2011；Pemov 等人，2018）。我们的提案将解决关键的未满足问题该领域的需求，包括更好的 ANF 体外和体内模型以及关键漏洞的识别这些细胞。为了提供临床前测试和预防 ANF 到 MPNST 发展的模型，我们沙漠刺猬 (Dhh)-Cre 驱动的 Nf1 双等位基因缺失与 Cdkn2a 杂合缺失相结合，创建一个独特的可移植 ANF 模型，该模型在先前存在的神经纤维瘤中发育（Chaney 等人，已提交）。我们还结合了 Dhh-Cre 驱动的 Nf1 和 Pten 双等位基因删除，产生了快速发展的围产期 ANF 样病变（Keng 等，2012）。来自 Dhh-Cre 的 ANF；Nf1fl/fl；Cdkn2a+/- 移植小鼠皮下注射到免疫功能低下的宿主中，在延迟后生长，提供了更快速、更容易处理的、转换系统。我们计划在这些模型中进行完整的转录组和外显子组分析（目标 1a），并且通过识别和验证小鼠体内改变的细胞群和标记来进一步研究该模型使用新的图像分析在未受干扰的组织切片中分析人类 PNF、ANF 和 MPNST 称为 CO-Detection by IndEXing 的方法（目标 1b）。免疫环境的调节正在增强用于治疗。因此，我们将定义神经微环境和免疫系统的影响从 ANF 进展到 MPNST（目标 1c）。为了识别 ANF 漏洞，我们完成了药物和基于 CRISPR 的基因合成致死率筛选同基因永生化人雪旺细胞（NF1）野生型或通过基因编辑使 NF1 功能丧失突变纯合。候选药物抑制 PP2A 的药物以及药物筛选工作中的其他新靶标将在 ANF 中测试其效果就像体外细胞（目标 2a）一样，如果成功的话，也可以在我们独特的 GEMM 中（目标 2b）。同样，我们的遗传筛选工作将用于定义体外（目标 2c）和体内（目标 2d）测试的其他漏洞。