A Novel Vector Platform to Actualize T Cell Modification In Vivo

一种在体内实现 T 细胞修饰的新型载体平台

• 批准号: 10663022
• 负责人: David Terry Curiel
• 金额: $ 42.76万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-04 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10663022
• 关键词: Ablation; Address; Adenoviruses; Advanced Development; BCAR1 gene; Capsid; Cells; Chimera organism; Chronic; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Dependovirus; Development; Disease; Engineering; Fiber; Fostering; Gene Delivery; Gene Transfer; Generations; Genes; Genetic; Genome; Human; Immunotherapy; In Situ; Intervention; Liver; Locales; Malignant Neoplasms; Methods; Modification; Mus; Organ; Patients; Serotyping; Simian Adenoviruses; Single-Stranded DNA; Sum; Surface; System; T-Lymphocyte; Techniques; Technology; Time; Tissues; Transgenes; Tropism; Viral; Virus; Virus Diseases; base editing; chimeric antigen receptor; chimeric antigen receptor T cells; cost; design; flexibility; gene delivery system; improved; in vivo; mouse model; neoplastic; novel; novel strategies; particle; practical application; repaired; technology platform; therapeutic genome editing; therapy outcome; transduction efficiency; vector

ABSTRACT To address the need for improved gene editing delivery systems, we propose constructing a chimeric “AdAAV” vector consisting of an adenovirus (Ad) with multiple adeno-associated viruses (AAVs) conjugated to its capsid's surface. We plan to employ the SpyTag/SpyCatcher technology to conjugate AAVs onto the Ad capsid. Of note in this regard, Ads are able to selectively target certain tissues (such as the liver in the case of huAd5) with high transduction efficiencies. Furthermore, our group has been able to engineer Ad fibers to facilitate tissue-specific targeting of T cells. In addition, by choosing an appropriate AAV serotype, and/or through capsid engineering AAVs can also selectively target desired tissues as well. AdAAVs may therefore provide superior targeting through the combined effects of both engineered Ad fiber and AAV capsid (which are matched to target the same tissue type). As a proof-of-concept, we plan to target T cells with AdAAVs. In addition, because AAVs carry single-stranded DNA, they can provide single-stranded donor templates, which are known to enhance the editing efficiency of homology directed repair (HDR). Within an AdAAV, the Cas protein can be encoded by the Ad genome while the single-stranded DNA template can be embodied within the genome of the AAVs. By virtue of the advantages of single-stranded donor templates for HDR, and by virtue of the high copy number of donor templates carried by the multiple AAVs associated with each Ad, this design might substantially increase editing efficiency. Finally, the AdAAV would possess a large packaging capacity since it would consist of a sum of the Ad's capacity (which is already high) and the AAV's capacity. Due to these factors, we suggest that AdAAVs may form a powerful and versatile new delivery system for gene editing therapies which overcomes many of the limitations associated with existing approaches. Our highly original AdAAV delivery system will greatly enhance the versatility of existing CRISPR-Cas gene editing therapies by circumventing several key obstacles to their broader applicability. The design features of AdAAV will potentially make it an ideal vector by which to address the challenges of gene editing delivery and thereby broadly enhance the general feasibility of gene editing-based therapies.

抽象的 为了满足改进基因编辑传递系统的需求，我们建议构建嵌合“AdAAV” 由腺病毒 (Ad) 和与其衣壳缀合的多个腺相关病毒 (AAV) 组成的载体 表面。我们计划采用 SpyTag/SpyCatcher 技术将 AAV 结合到 Ad 衣壳上。值得注意的是 在这方面，广告能够选择性地针对某些组织（例如 huAd5 的肝脏），具有高 转导效率。此外，我们的团队已经能够设计Ad纤维以促进组织特异性 T 细胞的靶向。此外，通过选择合适的 AAV 血清型和/或通过衣壳工程 AAV 还可以选择性地靶向所需的组织。因此，AdAAV 可能提​​供卓越的靶向性 通过工程化 Ad 纤维和 AAV 衣壳（它们与相同的目标相匹配）的综合作用 组织类型）。作为概念验证，我们计划用 AdAAV 靶向 T 细胞。此外，由于 AAV 携带 单链 DNA，他们可以提供单链供体模板，已知可以增强编辑 同源定向修复（HDR）的效率。在 AdAAV 中，Cas 蛋白可以由 Ad 编码 基因组，而单链 DNA 模板可以包含在 AAV 的基因组内。凭借 HDR单链供体模板的优点，并凭借供体的高拷贝数 与每个广告关联的多个 AAV 携带的模板，这种设计可能会大大增加编辑量 效率。最后，AdAAV 将拥有很大的包装容量，因为它将包含以下总和： 广告的容量（已经很高）和 AAV 的容量。由于这些因素，我们建议 AdAAV 可能会形成一个强大且多功能的基因编辑疗法新传递系统，克服许多问题 与现有方法相关的局限性。我们高度原创的AdAAV传输系统将大大增强 现有 CRISPR-Cas 基因编辑疗法的多功能性，克服了其应用的几个关键障碍 更广泛的适用性。 AdAAV 的设计特征将有可能使其成为解决以下问题的理想载体： 基因编辑交付的挑战，从而广泛增强基于基因编辑的总体可行性 疗法。