通过条形码扩增子测序绘制小鼠中常用腺相关病毒(AAV)变体的给药途径依赖性转导图谱。
Mapping administration route-dependent transduction profiles of commonly used AAV variants in mice by barcode amplicon sequencing.
作者信息
Abele Sarah, Alanis-Lobato Gregorio, Oti Martin, Rust Werner, Blazevic Dragica, Danner-Liskus Jenny, Mayer Christine, Zimmermann Gudrun, Zuckschwerdt Kai, Schönberger Tanja, Gross Peter, Lempp Charlotte, Michelfelder Stefan, Strobel Benjamin
机构信息
Global Drug Discovery Sciences, Boehringer Ingelheim Pharma GmbH & Co. KG, 88397 Biberach, Germany.
Global Computational Biology & Digital Sciences, Boehringer Ingelheim Pharma GmbH & Co. KG, 88397 Biberach, Germany.
出版信息
Mol Ther Methods Clin Dev. 2025 Apr 14;33(2):101468. doi: 10.1016/j.omtm.2025.101468. eCollection 2025 Jun 12.
The tissue transduction profiles and transgene expression efficiencies of adeno-associated viruses (AAVs) depend not only on the utilized capsid and dose but also on the administration route. Yet, despite the plethora of available natural and capsid-engineered variants, a comprehensive evaluation of the administration route dependency of AAV tropism has been lacking so far. Therefore, we here compared transduction and transgene expression profiles for 34 well-known AAV capsids following intravenous (i.v.) and intraperitoneal (i.p.) injection in male C57BL/6 mice by multiplexed biodistribution analyses based on AAV genome-barcoding. Readout on viral genome and transcript level confirmed pronounced liver targeting by most AAV variants after i.v. administration, as well as known tissue tropism for benchmark capsids (e.g., AAV-PHP.eB: brain and AAV2-ESGHGYF: lung). In contrast, i.p. administration generally decreased liver targeting, while concurrently increasing expression in other abdominal organs in a capsid-specific fashion. For example, AAV6.2 and AAV-DJ, which showed almost exclusive liver transduction after i.v. administration, displayed differential biodistribution profiles with enhanced expression in the diaphragm, adipose tissue, and pancreas when administered intraperitoneally. In summary, our data guide study design by enabling the selection of optimal vector and administration route combinations for refined tissue targeting approaches in preclinical experiments.
腺相关病毒(AAV)的组织转导谱和转基因表达效率不仅取决于所使用的衣壳和剂量,还取决于给药途径。然而,尽管有大量可用的天然和衣壳工程变体,但迄今为止,对AAV嗜性的给药途径依赖性缺乏全面评估。因此,我们在此通过基于AAV基因组条形码的多重生物分布分析,比较了34种著名AAV衣壳在雄性C57BL/6小鼠静脉内(i.v.)和腹腔内(i.p.)注射后的转导和转基因表达谱。病毒基因组和转录水平的读数证实,静脉注射后大多数AAV变体对肝脏有明显的靶向作用,以及已知的基准衣壳的组织嗜性(例如,AAV-PHP.eB:脑和AAV2-ESGHGYF:肺)。相比之下,腹腔注射通常会降低肝脏靶向性,同时以衣壳特异性方式增加其他腹部器官中的表达。例如,静脉注射后几乎只在肝脏转导的AAV6.2和AAV-DJ,腹腔注射时显示出不同的生物分布谱,在膈肌、脂肪组织和胰腺中的表达增强。总之,我们的数据通过为临床前实验中精细的组织靶向方法选择最佳载体和给药途径组合,为研究设计提供了指导。