Incorporation of Novel MADR-GESTALT Technology into UCLA SPORE in Brain Cancer

将新型 MADR-GESTALT 技术纳入 UCLA SPORE 治疗脑癌

• 批准号: 10271986
• 负责人: Linda M Liau
• 金额: $ 21.4万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-11 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10271986
• 关键词: Active Immunotherapy; Address; Adult; Affect; Alternative Splicing; Animal Model; Antigen-Presenting Cells; Applications Grants; Area; Award; Bar Codes; Brain Neoplasms; Brain Stem; CRISPR/Cas technology; Cancer Biology; Cancer Patient; Childhood Brain Neoplasm; Childhood Glioblastoma; Childhood Glioma; Clinic; Clinical Research; Communities; Cytotoxic T-Lymphocytes; Data; Development; Diagnosis; Diagnostic; Disease; Electroporation; Emerging Technologies; Enhancement Technology; Evolution; Funding; Gene Expression; Gene Transfer Techniques; Genes; Genetic; Genetic Heterogeneity; Genetic Predisposition to Disease; Genetic Variation; Glioblastoma; Glioma; Goals; Heterogeneity; Histones; Human; Immunocompetent; Immunotherapeutic agent; Immunotherapy; International; Lymphocyte Activation; Malignant Glioma; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Methodology; Modeling; Mosaicism; Mus; Mutation; Nature; Oncogenes; Parents; Pathogenicity; Patients; Perinatal; Physiologic pulse; Pre-Clinical Model; Predisposition; Primary Brain Neoplasms; Prognosis; Recurrence; Research; Research Project Grants; Safety; Somatic Mutation; System; T-Cell Receptor; Techniques; Technology; Transgenes; Translational Research; Translations; Tumor Subtype; Vaccination; Validation; Variant; anti-tumor immune response; anticancer research; career; checkpoint inhibition; checkpoint modulation; combinatorial; de novo mutation; disease heterogeneity; effective therapy; efficacy evaluation; exome sequencing; flexibility; genome editing; genome sequencing; genomic locus; immunoregulation; improved; in vivo; in vivo Model; innovation; insight; loss of function mutation; mouse model; mutant; neoantigens; novel; novel strategies; pediatric patients; pre-clinical; pressure; programs; receptor binding; recombinase; recombinase-mediated cassette exchange; stem cells; therapy resistant; tool; translational research program; tumor; tumorigenesis

ABSTRACT (Overall) Glioblastoma, the most common primary brain tumor in adults, is one of the most lethal of all human cancers. This disease requires innovative approaches and more effective treatments. Treatment resistance presents a fundamental barrier, and given the heterogeneous nature of malignant gliomas, a combination of diverse approaches will likely be needed to overcome it. To achieve this, we need a better understanding of the genetic diversity and heterogeneity of the disease, as well as better pre-clinical animal models that recapitulate human glioma heterogeneity. A state-of-the-art methodology—mosaic analysis with dual recombinases (MADR) and genome editing of synthetic target arrays for lineage tracing (GESTALT)—addresses this need. MADR provides a flexible means for single-copy somatic transgenesis (or mutation with CRISPR/Cas9). MADR-GESTALT provides a suite of tools to study the emergent heterogeneity in cancer formation and recurrence by providing a consistent, single-copy, genetic framework for the expression of multiple “personalized” patient driver genes. Perinatal electroporation is used to deliver genes to brain stem and progenitor cells, and to avoid multiple copy insertion, a new technique for single-copy transgene insertion in vivo was developed. Mosaic Analysis with Dual Recombinases (MADR) employs dual recombinase-mediated cassette exchange for high efficiency insertion of transgenes to a single genetic locus. To model loss-of-function mutations, CRISPR/Cas9-mediated gene editing is also incorporated into the MADR-GESTALT system. Several areas of MADR can be expanded and leveraged to enhance specific projects within the UCLA Brain Cancer SPORE, which, in addition, will independently validate the utility of MADR-GESTALT for the wider cancer research community. This proposal utilizes an IMAT-funded technology that enhances three ongoing projects within our SPORE program: 1) Active immunotherapy combined with checkpoint modulation for glioblastoma. MADR will be used to develop syngeneic immunocompetent mice with gliomas and GESTALT will be used to evaluate barcode diversity with and without treatment to further understand tumor evolution under immunotherapeutic pressure. 2) Genetic susceptibility in pediatric glioma development. MADR-GESTALT will be used to establish a moderate-throughput, high-fidelity, patient-specific in vivo modeling platform to understand pathogenicity of novel germline variants, their effects on gene expression, and their contribution to pediatric high-grade glioma susceptibility. 3) Adaptive immunotherapy to target the H3.3G34 mutation in glioblastoma. MADR-GESTALT models will be used to examine mechanisms by which particular histone mutations affect oncogenesis and immunotherapy for G34R mutant glioblastoma.

摘要(总体) 胶质母细胞瘤是成人最常见的原发脑瘤，也是人类所有癌症中最致命的癌症之一。 这种疾病需要创新的方法和更有效的治疗方法。治疗抗性呈现出一种 基本障碍，并鉴于恶性胶质瘤的异质性，结合了多种 可能需要一些方法来克服它。要做到这一点，我们需要更好地了解基因 疾病的多样性和异质性，以及更好的临床前动物模型 胶质瘤的异质性。一种最新的方法学--双重组酶嵌合分析 用于谱系追踪的合成目标阵列的基因组编辑(格式塔)--满足这一需求。MADR提供 单拷贝体细胞转基因的灵活方法(或CRISPR/Cas9突变)。马德尔-格式塔 提供了一套工具来研究癌症形成和复发中的紧急异质性，通过提供 一致的、单拷贝的、用于表达多个“个性化”患者驾驶基因的遗传框架。 围产期电穿孔被用来将基因输送到脑干和祖细胞，并避免多次复制 建立了一种新的体内单拷贝转基因插入技术。具有对偶的马赛克分析 重组酶(MADR)利用双重重组酶介导盒交换高效插入 转基因到一个单一的遗传位点。为了模拟功能丧失突变，CRISPR/Cas9介导的基因编辑 也被纳入MADR-格式塔系统。MADR的几个领域可以扩大和利用 加强加州大学洛杉矶分校脑癌孢子内的具体项目，此外，这些项目将独立验证 MADR-格式塔在更广泛的癌症研究社区中的效用。这项提议利用了IMAT资助的 加强我们孢子计划中正在进行的三个项目的技术：1)主动免疫疗法 联合检查点调制治疗胶质母细胞瘤。MADR将用于开发同基因 免疫活性的胶质瘤小鼠和格式塔小鼠将被用来评估有无条形码的多样性 进一步了解免疫治疗压力下肿瘤的演变。2)遗传易感性 儿童脑胶质瘤的发展。MADR-格式塔将用于建立中等吞吐量、高保真、 患者特定的体内建模平台，以了解新的生殖系变异的致病性，它们对 基因表达及其对儿童高级别胶质瘤易感性的贡献。3)适应性免疫疗法 目的：针对胶质母细胞瘤中H3.3G34突变。MADR-格式塔模型将被用来检验机制 通过特定的组蛋白突变影响G34R突变的胶质母细胞瘤的肿瘤发生和免疫治疗。