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: 10709378
• 负责人: Linda M Liau
• 金额: $ 23.56万
• 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 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10709378
• 关键词: Acceleration; Address; Adult; Affect; Alternative Splicing; Animal Model; Antigen Targeting; Apoptosis; Apoptotic; Applications Grants; Area; Award; BCL2L1 gene; Bar Codes; Brain Neoplasms; Brain Stem; CRISPR/Cas technology; CSF1R gene; Cancer Biology; Cancer Patient; Childhood Brain Neoplasm; Childhood Glioblastoma; Clinic; Clinical; Communities; Cytotoxic T-Lymphocytes; DNA Damage; Data; Development; Diagnosis; Disease; Electroporation; Emerging Technologies; Enhancement Technology; Evolution; Funding; Gene Transfer Techniques; Genes; Genetic; Genetic Heterogeneity; Genetic Variation; Glioblastoma; Glioma; Goals; Heterogeneity; Histones; Human; Immune response; Immunocompetent; Immunotherapeutic agent; Immunotherapy; International; Lymphocyte Activation; Malignant Glioma; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Methodology; Modeling; Mus; Mutation; Myeloid-derived suppressor cells; Nature; Newly Diagnosed; Oncogenes; Parents; Patients; Perinatal; Pharmaceutical Preparations; Primary Brain Neoplasms; Prognosis; Recurrence; Research; Research Project Grants; Resistance; Safety; Somatic Mutation; System; T-Cell Receptor; Techniques; Technology; Testing; Transgenes; Translational Research; Translations; Tumor Subtype; Tumor-infiltrating immune cells; Vaccination; Validation; Variant; anti-tumor immune response; anticancer research; combinatorial; disease heterogeneity; drug candidate; effective therapy; flexibility; genome editing; genomic locus; improved; in vivo; innovation; insight; loss of function mutation; mosaic analysis; mouse model; mutant; neoantigens; neoplasm immunotherapy; novel; novel strategies; novel therapeutics; pre-clinical; pressure; programmed cell death protein 1; programs; receptor binding; recombinase; recombinase-mediated cassette exchange; resistance mechanism; small molecule inhibitor; stem cells; targeted treatment; therapy resistant; tool; translational goal; 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 recombinase-mediated cassette exchange (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) Targeting immunotherapy-resistance with DC vaccination and PD-1/CSF-1R inhibition. 2) Overcoming drug-induced resistance to intrinsic apoptosis in 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 and small molecule inhibitor pressure. 3) Adaptive immunotherapy to target the H3.3G34 mutation in pediatric 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） 胶质母细胞瘤是成年人中最常见的原发性脑肿瘤，是所有人类癌症中最致命的一种。 该疾病需要创新的方法和更有效的治疗方法。治疗抗性呈现 基本障碍，并鉴于恶性神经胶质瘤的异质性，潜水员的结合 可能需要采取方法来克服它。为了实现这一目标，我们需要更好地了解通用 疾病的多样性和异质性，以及概括人类的临床前动物模型 胶质瘤异质性。一种最先进的方法论 - 双重重组酶介导的摩萨克分析 盒式磁带阵列（Gestalt）的合成目标阵列（Gestalt）的盒式盒式交换（MADR）和基因组编辑 - 解决了这一需求。 MADR为单拷贝躯体翻译提供了灵活的手段（或与 CRISPR/CAS9）。 Madr-Gestalt提供了一套工具来研究癌症的新兴异质性 通过提供一致的单拷贝，遗传框架来表达多重的形成和复发 “个性化”患者基因。围产期电穿孔用于将基因传递给脑干和 祖细胞，并避免多个复制插入，这是一种单拷贝变换插入体内的新技术 双重重组酶（MADR）员工双重重组酶介导的马赛克分析 盒式交换以高效插入单个遗传基因座的效率。建模功能丧失 突变，CRISPR/CAS9介导的基因编辑也已纳入MADR-GESTALT系统。一些 可以扩展和利用MADR的区域，以增强UCLA脑癌内的特定项目 孢子此外，它将独立验证Madr-Gestalt对更广泛癌症的实用性 研究社区。该提案利用了IMAT资助的技术，可以增强三个正在进行的项目 在我们的孢子计划中：1）针对DC疫苗和PD-1/CSF-1R的免疫治疗抗性 抑制。 2）克服胶质母细胞瘤中药物诱导的对固有凋亡的抗性。马德尔将习惯 使用神经胶质瘤和格式塔开发合成性免疫能力小鼠将用于评估条形码 有或没有治疗的多样性，以进一步了解免疫治疗和小的肿瘤演变 分子抑制剂压力。 3）适应性免疫疗法以针对小儿H3.3G34突变 胶质母细胞瘤。 Madr-Gestalt模型将用于检查特定组蛋白的机制 突变会影响G34R突变胶质母细胞瘤的肿瘤发生和免疫疗法。

项目成果

The clinical trials landscape for glioblastoma: is it adequate to develop new treatments?

• DOI: 10.1093/neuonc/noy027
• 发表时间: 2018-08-01
• 期刊: NEURO-ONCOLOGY
• 影响因子: 15.9
• 作者: Vanderbeek, Alyssa M.;Rahman, Rifaquat;Alexander, Brian M.
• 通讯作者: Alexander, Brian M.

Has Toxicity Testing Moved into the 21st Century? A Survey and Analysis of Perceptions in the Field of Toxicology.

• DOI: 10.1289/ehp1435
• 发表时间: 2017-08-30
• 期刊: Environmental health perspectives
• 影响因子: 10.4
• 作者: Zaunbrecher V;Beryt E;Parodi D;Telesca D;Doherty J;Malloy T;Allard P
• 通讯作者: Allard P

Molecular profiling of stem cell-derived retinal pigment epithelial cell differentiation established for clinical translation.

• DOI: 10.1016/j.stemcr.2022.05.005
• 发表时间: 2022-06-14
• 期刊: STEM CELL REPORTS
• 影响因子: 5.9
• 作者: Petrus-Reurer, Sandra;Lederer, Alex R.;Baque-Vidal, Laura;Douagi, Iyadh;Pannagel, Belinda;Khven, Irina;Aronsson, Monica;Bartuma, Hammurabi;Wagner, Magdalena;Wrona, Andreas;Efstathopoulos, Paschalis;Jaberi, Elham;Willenbrock, Hanni;Shimizu, Yutaka;Villaescusa, J. Carlos;Andre, Helder;Sundstrom, Erik;Bhaduri, Aparna;Kriegstein, Arnold;Kvanta, Anders;La Manno, Gioele;Lanner, Fredrik
• 通讯作者: Lanner, Fredrik

Cytokines Produced by Dendritic Cells Administered Intratumorally Correlate with Clinical Outcome in Patients with Diverse Cancers.

• DOI: 10.1158/1078-0432.ccr-17-2707
• 发表时间: 2018-08-15
• 期刊: Clinical cancer research : an official journal of the American Association for Cancer Research
• 作者: Subbiah V;Murthy R;Hong DS;Prins RM;Hosing C;Hendricks K;Kolli D;Noffsinger L;Brown R;McGuire M;Fu S;Piha-Paul S;Naing A;Conley AP;Benjamin RS;Kaur I;Bosch ML
• 通讯作者: Bosch ML

Perfusion and diffusion MRI signatures in histologic and genetic subtypes of WHO grade II-III diffuse gliomas.

• DOI: 10.1007/s11060-017-2506-9
• 发表时间: 2017-08
• 期刊: Journal of neuro-oncology
• 影响因子: 3.9
• 作者: Leu K;Ott GA;Lai A;Nghiemphu PL;Pope WB;Yong WH;Liau LM;Cloughesy TF;Ellingson BM
• 通讯作者: Ellingson BM