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 为了满足改善基因编辑输送系统的需求，我们提出构建嵌合“ Adaav” 由腺病毒（AD）组成的矢量，以及与帽子的多种相关病毒（AAV）的媒介 表面。我们计划采用Spytag/Spycatcher技术将AAV偶联到AD Capsid上。值得注意 在这方面，广告能够选择性地靶向某些组织（例如HUAD5的肝脏） 转导功能。此外，我们的小组已经能够设计出广告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Capsid Engineering AAV也可以选择性地靶向所需的组织。因此，Adaavs可以提供卓越的目标 通过工程广告纤维和AAV帽的综合效果（它们与目标相同 组织类型）。作为概念验证，我们计划用ADAAV靶向T细胞。另外，因为AAV携带 单链DNA，它们可以提供单链供体模板，已知可以增强编辑 同源性修复效率（HDR）。在ADAAV中，CAS蛋白可以由AD编码 基因组虽然单链DNA模板可以体现在AAV的基因组中。依靠 HDR的单链供体模板的优点，以及依赖于高拷贝数的捐助者 由与每个广告相关的多个AAV携带的模板，此设计可能会大大增加编辑 效率。最后，阿达夫（Adaav）将具有较大的包装能力，因为它由 AD的容量（已经很高）和AAV的容量。由于这些因素，我们建议Adaavs 可能形成一个强大而多才多艺的新输送系统，用于基因编辑疗法，以克服许多 与现有方法相关的限制。我们高度原始的ADAAV交付系统将极大地增强 现有的CRISPR-CAS基因编辑疗法的多功能性通过规避了几个关键障碍 更广泛的适用性。 Adaav的设计特征有可能使其成为理想的向量 基因编辑递送的挑战，从而广泛提高了基于基因编辑的一般可行性 疗法。