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：
10247078
负责人：
DAVID ANDREW LARGAESPADA
金额：
$ 58.29万
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/10247078
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).

项目摘要/摘要神经纤维瘤病1型（NF1）综合征是一种常染色体主导癌症癌综合征大多数患者在全球每3,500人中每3,500人中〜1 神经纤维瘤。丛状神经纤维瘤构成了终生的毁容，发病率和死亡率的来源并有可能转化为恶性周围神经鞘肿瘤（MPNST），一种侵略性的柔软组织肉瘤。实际上，大约15％的NF1患者的预后较差，通常在生命的第二或第三十年。 MPNST的治疗选择仅限于复杂的手术程序以及迄今为止的经典化学疗法以及分子靶向疗法的效率有限。我们迫切需要MPNST和方法的新治疗选择，以防止它们开发。它最近发现，丛状神经纤维瘤通过中间体“非典型”发展到mpnst 神经纤维瘤（ANF）在70％的肿瘤中显示CDKN2A的杂合损失编码P16INK4A和P14ARF（Beert等，2011; Pemov等，2018）。我们的建议将解决关键该领域的需求，包括更好的体外和体内模型，以及对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开发的模型，我们沙漠刺猬（DHH） - Cre驱动的NF1驱动的双重缺失，杂合损失CDKN2A，创建一个在现有神经纤维瘤中发展的独特的可移植ANF的模型（Chaney等，提交）。我们还将NF1和PTEN的DHH-CRE驱动双重缺失结合在一起，产生了迅速发展围产期类似ANF的病变（Keng等，2012）。 dhh-cre; nf1fl/fl; cdkn2a +/-小鼠接枝的anf 延迟后，皮下介入免疫功能低下的宿主会生长，提供更快，更可行的宿主转换系统。我们在这些模型（AIM 1A）中计划完整的转录组和外显子组分析，以及进一步研究模型，通过识别和验证小鼠中改变的细胞群和标记使用新的图像分析，在不受干扰的组织切片中，人类PNF，ANF和MPNST 通过索引称为共检测的方法（AIM 1B）。免疫环境的调节正在增加使用的理论。因此，我们将定义神经微环境和免疫系统的影响关于从ANF到MPNST的进展（AIM 1C）。为了识别ANF漏洞，我们已经完成了药物，并且基于CRISPR的遗传合成致死性筛选是NF1 野生型或使用基因编辑的NF1丧失功能突变的纯合子。候选药物抑制PP2A以及药物筛查工作中的其他新靶标，将测试其在ANF- 例如体外细胞（AIM 2A），并且在我们独特的宝石（AIM 2B）中成功时。同样，我们的遗传筛查工作将用于定义在体外测试的其他漏洞（AIM 2C）和体内（AIM 2D）。