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

• 批准号: 10653687
• 负责人: DAVID ANDREW LARGAESPADA
• 金额: $ 57.18万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 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/10653687
• 关键词: Address; Alleles; Allografting; 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; Heterozygote; Human; Image Analysis; Immune; Immune system; Immunocompromised Host; In Vitro; Lesion; Lethal Genes; Life; Loss of Heterozygosity; 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; Surgical complication; Syndrome; System; Testing; Therapeutic Uses; Tissues; Wild Type Mouse; autosome; 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

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。

项目成果

Modeling human cancer predisposition syndromes using CRISPR/Cas9 in human cell line models.

• DOI: 10.1002/gcc.23140
• 发表时间: 2023-09
• 期刊: GENES CHROMOSOMES & CANCER
• 影响因子: 3.7
• 作者: Draper, Garrett M.;Panken, Daniel J.;Largaespada, David A.
• 通讯作者: Largaespada, David A.

Selumetinib normalizes Ras/MAPK signaling in clinically relevant neurofibromatosis type 1 minipig tissues in vivo.

• DOI: 10.1093/noajnl/vdab020
• 发表时间: 2021-01
• 期刊: Neuro-oncology advances
• 作者: Osum SH;Coutts AW;Duerre DJ;Tschida BR;Kirstein MN;Fisher J;Bell WR;Delpuech O;Smith PD;Widemann BC;Moertel CL;Largaespada DA;Watson AL
• 通讯作者: Watson AL