Development of double-targeted vectors for long-term vascular expression in vivo

开发用于体内长期血管表达的双靶向载体

• 批准号: 7653126
• 负责人: David Terry Curiel
• 金额: $ 58.68万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7653126
• 关键词: Adenovirus Vector; Adenoviruses; Antibodies; Biology; Blood Vessels; Capsid; Cardiovascular Diseases; Cell surface; Clinical; Clinical Trials; Data; Development; Disease; Disseminated Malignant Neoplasm; Dose-Limiting; Endothelium; Engineering; Gene Delivery; Gene Expression; Gene Transduction Agent; Generations; Genes; Hepatic; Hepatotoxicity; Human; Hypoxia; Injection of therapeutic agent; Liver; Lung; Mediating; Methodology; Property; Pulmonary Hypertension; Recombinant Antibody; Rodent Model; Safety; System; Technology; Therapeutic; Tissues; Translations; Vascular Diseases; base; clinical application; clinically relevant; disease phenotype; gene therapy; gutless adenoviral vector; immunogenic; immunogenicity; in vivo; large scale production; practical application; public health relevance; pulmonary arterial hypertension; transgene expression; vector

DESCRIPTION (provided by applicant): To create a clinically relevant vector for vascular-based diseases, we must advance three pivotal efforts. First, it is critical to formally correct an established disease phenotype via systemic administration of a vector that de-targets the liver and thus reduces adenovirus (Ad) associated liver toxicity. Second, development of a safer, less immunogenic Ad vector capable of long-term gene expression will be required to realize human clinical translation of a gene therapy for diseases involving endothelium. To this end, we will combine our current targeting technology with "gutless" Ad vectors, known to provide long-term gene expression in vivo as a result of their reduced immunogenicity. Finally, to further increase the clinical utility of our vector, we propose the development of a single component targeted Ad vector compatible with large scale production and FDA approval. In this regard, we have recently developed the methodology to incorporate functional single chain antibody (scFv) targeting moieties into the Ad capsid resulting in highly selective gene delivery to specific cell surface markers. This key technological development will allow us to extend our established targeting paradigm to a single component vector with genetically incorporated targeting. We hypothesize that the combination of these technologies will allow the realization of a clinically relevant gene therapy vector for diseases requiring systemic injection and significant pulmonary endothelium involvement. Our specific aims are thus: Specific Aim 1: To evaluate the ability of double-targeted Ad vectors to correct disease involving the pulmonary endothelium. Specific Aim 2: To create targeted gutless Ad vectors capable of long term gene expression in the pulmonary endothelium. Specific Aim 3: To create a single-component, targeted gutless Ad employing genetically incorporated recombinant antibodies to target the pulmonary endothelium. The successful completion of these specific aims will provide important and timely data regarding the efficacy of endothelial targeted gene therapy and lay the groundwork for other endothelial based diseases. Further, our studies will clearly establish therapeutic gain associated with decreased vector immunogenicity and long term gene expression. Finally, the studies proposed herein will provide a critical step toward a safer, single component vector compatible with large-scale production and clinical translation. PUBLIC HEALTH RELEVANCE: Adenovirus (Ad) based vectors are widely used in clinical trials due to their well characterized biology and positive safety profile. However, the majority of clinical applications employing Ad based vectors utilize local administration due to limitations associated with first generation Ad vectors. In this proposal, we seek to engineer a safer, less immunogenic Ad vector capable of long-term gene expression.

描述（由申请人提供）：为了创建针对血管疾病的临床相关载体，我们必须推进三项关键工作。首先，通过全身施用一种载体来正式纠正已确定的疾病表型至关重要，这种载体可以使肝脏脱靶，从而减少腺病毒（Ad）相关的肝脏毒性。其次，需要开发一种更安全、免疫原性较低、能够长期基因表达的Ad载体，以实现针对内皮疾病的基因疗法的人体临床转化。为此，我们将把我们目前的靶向技术与“无胆”Ad载体相结合，众所周知，由于其免疫原性降低，可以在体内提供长期基因表达。最后，为了进一步提高我们的载体的临床实用性，我们建议开发一种与大规模生产和 FDA 批准兼容的单组分靶向 Ad 载体。在这方面，我们最近开发了一种方法，将功能性单链抗体 (scFv) 靶向部分整合到 Ad 衣壳中，从而将高度选择性的基因递送到特定的细胞表面标记。这一关键技术的发展将使我们能够将我们已建立的靶向范式扩展到具有基因整合靶向的单组分载体。我们假设这些技术的结合将能够实现临床相关的基因治疗载体，用于治疗需要全身注射和显着肺内皮参与的疾病。因此，我们的具体目标是： 具体目标 1：评估双靶向 Ad 载体纠正肺内皮疾病的能力。具体目标 2：创建能够在肺内皮细胞中长期表达基因的靶向无肠 Ad 载体。具体目标 3：利用基因整合的重组抗体来靶向肺内皮，创建单组分、靶向无肠广告。这些具体目标的成功完成将为内皮靶向基因治疗的功效提供重要且及时的数据，并为其他基于内皮的疾病奠定基础。此外，我们的研究将清楚地确定与载体免疫原性降低和长期基因表达相关的治疗增益。最后，本文提出的研究将为实现与大规模生产和临床转化兼容的更安全、单组分载体迈出关键一步。 公共卫生相关性：基于腺病毒 (Ad) 的载体因其良好的生物学特征和积极的安全性而广泛用于临床试验。然而，由于第一代 Ad 载体的局限性，大多数使用基于 Ad 的载体的临床应用均采用局部给药。在本提案中，我们寻求设计一种更安全、免疫原性更低、能够长期基因表达的 Ad 载体。