Targeting glioblastoma stem-like cells with custom-designed viral vectors

用定制设计的病毒载体靶向胶质母细胞瘤干细胞

• 批准号: 10021240
• 负责人: Darin J Falk
• 金额: $ 15.63万
• 依托单位: LACERTA THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10021240
• 关键词: Address; Adult; Apoptotic; Astrocytes; Biological; Biological Assay; Biology; CD44 gene; Capsid; Cell surface; Cells; Clinical Trials; Collaborations; Custom; DNA cassette; Data; Dependovirus; Development; Directed Molecular Evolution; Environment; Exhibits; Expression Profiling; Florida; Flow Cytometry; Gene Expression; Genetic; Genetic Determinism; Genomics; Glioblastoma; Goals; Heterogeneity; In Vitro; Individual; Injections; Inter-tumoral heterogeneity; Lead; Libraries; Malignant - descriptor; Malignant neoplasm of brain; Modeling; Molecular; Pathway interactions; Patients; Phase; Platelet Factor 4; Population; Population Heterogeneity; Preclinical Testing; Probability; Process; Radiation; Reagent; Recurrence; Regimen; Safety; Serotyping; Signal Pathway; Specificity; Stem cells; Systems Biology; Technology; Time; Toxic effect; Treatment Failure; Universities; Variant; Viral; Viral Vector; Xenograft Model; Xenograft procedure; adeno-associated viral vector; base; chemoradiation; clinical application; clinical development; combinatorial; computational platform; computerized tools; design; efficacy study; gene therapy; improved; in vivo; in vivo evaluation; interdisciplinary approach; knock-down; lead optimization; mouse model; new technology; novel; novel therapeutics; pharmacokinetics and pharmacodynamics; pre-clinical; preclinical efficacy; programs; research clinical testing; small hairpin RNA; stem-like cell; therapeutic development; tool; transduction efficiency; tumor; tumor progression; tumorigenesis; vector

Glioblastoma (GBM) is the most lethal form of adult brain cancers with a median survival of <15 months despite aggressive standard chemoradiation. GBM are formed by GBM stem-like cells (GSCs) - a major contributor to tumor recurrence and a natural focus for therapeutic development. There are two main reasons responsible for treatment failure: 1) high intra- and inter-tumor cellular and molecular heterogeneity with multiple subclones possessing distinct genetic determinants; 2) GSCs exhibit multiple redundant signaling pathways requiring simultaneous targeting of overlapping pathways. We have invented and biologically validated a novel tandem computational platform, GeneRep-nSCORE that integrates large-scale gene expression profiles with genomic changes to identify common founding alterations or master regulators of GSCs that span a large number, if not all, GSC subclones within and across GBM tumors. We discovered such a core set of four common master regulators in GCSs that are outstanding targets for clinical development. Expression of these four factors was sufficient to reprogram normal astrocytes to GSCs, whereas their depletion profoundly abrogated GSCs, and thus tumor development in vivo, in all eight lines of patient-derived GSCs of varied genetic and molecular backgrounds examined to date. The goal of this application is to develop a customized set of Adeno-associated virus (AAV)-based genetic tools to target the whole spectrum of GSCs (Phase I) for the purpose of delivering targeting constructs to deplete the four common master regulators responsible for malignant transformation and proliferation in GSCs (Phase II). The specific objectives of this proposal are: (i) using directed evolution and available combinatorial AAV capsid library, and for the first time, introducing a dynamic mode of administration of a library reagent over the time course of tumor progression, to greatly increase the probability of identifying novel AAV variants specifically targeting slow-, and fast-cycling GSCs in patient-derived xenograft models (PDX) (Phase I); (ii) To design and validate a panel of AAV vectors that express shRNAs targeting core master regulators of GSCs to identify leads for preclinical testing; (iii) To optimize modes of viral delivery, pharmacokinetics and pharmacodynamics parameters, and safety and toxicity in normal and PDX treated with lead targeting AAV cassettes; and (iv) Based on these results, tools and basic DMPK data created, to conduct preclinical efficacy studies in PDX treated with lead targeting AAV cassettes either alone or in combination with standard chemoradiotherapy (Phase II) to prepare for an investigative new drug application for clinical testing in patients with GBM, and for commercial development of this novel technology.

胶质母细胞瘤（GBM）是成年脑癌的最致命形式，中位生存期<15个月 尽管有积极的标准化学放疗。 GBM由GBM茎样细胞（GSC）形成 - 主要 肿瘤复发和治疗发育的自然重点。有两个主要原因 负责治疗失败：1）高肿瘤内和肿瘤间细胞和分子异质性 多个具有不同遗传决定因素的亚克隆； 2）GSC表现出多种冗余信号传导 需要同时定位重叠途径的途径。我们已经发明和生物学 验证了一个新颖的串联计算平台，Generep-Nscore，该平台积分大规模基因 具有基因组变化的表达曲线，以识别常见的基础改变或主要调节剂 跨越大量的GSC（如果不是全部）GSC子克隆和跨GBM肿瘤的GSC。我们发现了这样的 GCS中四个普通主调节器的核心集是临床开发的杰出目标。 这四个因素的表达足以将正常星形胶质细胞重新编程为GSC，而它们 在所有八行患者衍生的行中，耗竭深刻废除了GSC，因此在体内发育 迄今为止研究的各种遗传和分子背景的GSC。 该应用程序的目的是开发一组定制的腺相关病毒（AAV）的遗传 针对全GSC（I阶段）的工具，目的是将目标结构传递给 耗尽负责GSC中恶性转化和增殖的四个普通主调节器 （第二阶段）。该提案的具体目标是：（i）使用定向进化和可用组合 AAV CAPSID库，也是第一次引入库试剂的动态管理方式 在肿瘤进展的时间过程中，大大增加了识别新型AAV变体的可能性 专门针对患者衍生异种移植模型（PDX）（I阶段）中的慢速和快速循环GSC； （ii）到 设计和验证一组AAV矢量，以将GSC的核心主调节器表达为shRNA 识别临床前测试的潜在客户； （iii）优化病毒传递，药代动力学和 药效学参数以及用铅靶向AAV处理的正常和PDX的安全性和毒性 盒式磁带； （iv）基于这些结果，工具和基本DMPK数据，以执行临床前功效 用铅靶向AAV盒单独或与标准组合处理的PDX研究 化学放疗（II阶段）为患者的临床测试做准备 使用GBM，以及这项新技术的商业开发。