PROJECT 2: NF1-associated Glioblastoma

摘要(项目2) 我们的中心假设是，NF1突变的胶质瘤构成了一个独特的胶质瘤亚群，具有独特的 分子发病机制、肿瘤微环境和肿瘤细胞脆弱性。我们的方法包括 多种胶质瘤模型，包括GEMS、患者来源的异种移植(PDX)、原代培养和人类 肿瘤活检。我们将同时考虑生殖系和体细胞NF1沉默，并建立在 在靶向NF1相关的其他器官系统肿瘤发生方面取得了相当大的进展(项目1 3)最大限度地发挥这部作品的翻译效果。为了开发NF1基因特有的假说， 同基因宝石和衍生的GBM原代培养是我们发现细胞内在方面的首选 特性，以及免疫微环境和一线治疗测试。我们将利用鼠标 我们开发的通过灭活三个关键肿瘤自发形成完全外露的GBM的模型 这种疾病中的抑制基因(NF1、TP53和Pten)，并被设计成模拟生殖系和体细胞NF1 被驱使的疾病。在一组实验中，小鼠携带杂合性生殖系NF1突变(NF1-/FLOX或-/)，而 在另一种情况下，小鼠将只发生体细胞NF1突变(NF1/或NF1/FLOX)。发育成熟的小鼠 由于QKi、TP53和Pten基因突变引起的GBM(QPP小鼠)将作为这些疾病的对照 实验和代表NF1野生型GBM亚群。原代培养与肿瘤扩大 同种异体移植物将用于体外和体内的临床前研究。由于散发性GBM的遗传复杂性 在人类中不能在GEM中充分代表，我们将确认和推广PDX模型中的关键结果 基因类型不同，但NF1基因要么突变，要么野生型。NF1突变体和野生型 MSKCC开发的患者来源的原位异种移植(PDX)将与项目1相结合进行研究 以及3，测试NF1/RAS途径-量身定做的和该领域出现的其他疗法。人类肿瘤 样本与免疫渗透的评估最相关，将在独特的背景下进行检查 生殖系NF1-GBM患者的临床表现目标3将重点放在系统的临床注释上 与常规收集的详细的组织学和分子分析相匹配的生殖系NF1 GBM患者的 NF1相关GBM患者的福尔马林固定和石蜡包埋的肿瘤活检组织。这些研究 将与Core C(生物质学/病理学)合作执行，并将用于验证关键结果 关于TME的NF1相关特征。我们将与Core B(OMICS)合作执行 GEM、PDX和人体组织样品的详细基因组和蛋白质组特征分析 在NF1突变体GBM中特异的可操作的信号通路。将创建一个面向患者的门户网站来招聘 并系统地记录NF1相关的GBM的临床表现和病程，以便整合 GEM和PEX临床前模型中NF1驱动的胶质瘤的组织学、分子和临床数据 患有NF1 GBM的人。

ABSTRACT (Project 2) Our central hypothesis is that NF1 mutant gliomas constitute a distinct glioma subgroup with a unique molecular pathogenesis, tumor microenvironment, and tumor cell vulnerabilities. Our approach includes a variety of glioma models, including GEMs, patient-derived xenografts (PDX), primary cultures, and human tumor biopsies. We will consider both germline and somatic NF1 silencing, and also build upon the considerable recent progress with targeting NF1 associated tumorigenesis in other organ systems (Projects 1 and 3) to maximize the translational impact of this work. To develop NF1 genotype-specific hypotheses, syngeneic GEM and derived GBM primary cultures are our first choice for discovery aspects of cell intrinsic properties, as well as the immune microenvironment and first line therapeutic testing. We will exploit mouse models developed by us that spontaneously develop GBM with full penetrance by inactivating three key tumor suppressor genes in this disease (NF1, Tp53 and Pten) and engineered to mimic germline versus somatic NF1 driven disease. In one setting the mice harbor a heterozygous germline NF1 mutation (NF1-/flox or -/+), whereas in the other setting the mice will develop only somatic NF1 mutations (NF1flox/+ or NF1flox/flox). Mice that develop GBM due to mutations in the QKi, Tp53 and Pten genes (QPP mice) will serve as controls in these experiments and represent the subgroup of NF1 wild type GBM. Primary cultures and expanded tumor allografts will be used for in vitro and in vivo preclinical studies. Since the genetic complexity of sporadic GBM in humans is not adequately represented in GEM, we will confirm and extend key results in PDX models that vary in genotypes but for which the NF1 gene is either mutated or wildtype. NF1 mutant and NF1 wild type patient derived orthotopic xenografts (PDX) developed at MSKCC will be studied in alignment with Projects 1 and 3, to test NF1/Ras pathway-tailored and other therapies that have emerged in the field. Human tumor samples are most relevant for assessment of immune infiltration and will be examined in context of the unique clinical presentation of patients with germline NF1 GBM. Aim 3 will focus on the systematic clinical annotation of patients with germline NF1 GBM paired with detailed histologic and molecular analysis of routinely collected formalin-fixed and paraffin embedded tumor biopsies from people with NF1 associated GBM. These studies will be performed in collaboration with Core C (Biospecimen/Pathology) and will be used to validate key results regarding NF1 associated characteristics of the TME. We will collaborate with Core B (Omics) to perform detailed genomic and proteomic characterization of samples from GEM, PDX and human tissue to identify actionable signaling pathways specifically in NF1 mutant GBM. A patient-facing portal will be created to recruit and systemically document the clinical presentation and course of NF1 associated GBM to allow integration of histologic, molecular and clinical data for NF1 driven gliomas across GEM and PEX preclinical models and people with NF1 GBM.