Distinct infectivity and neutralization antibody responses in the highly homologous AAV Go.1 and AAV5
IntroductionGoat-derived adeno-associated virus (AAV) vectors, such as AAV Go.1, represent a novel platform for gene therapy due to their unique origin and potential advantages in transduction efficiency and immune evasion. However, their therapeutic potential and biological properties remain undere...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2025-04-01
|
| Series: | Frontiers in Medicine |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmed.2025.1554449/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849735687968915456 |
|---|---|
| author | Mei Li Haixiao Ma Yang Wu Yunling Gao Jie Wang Jie Wang Hanbing Wang |
| author_facet | Mei Li Haixiao Ma Yang Wu Yunling Gao Jie Wang Jie Wang Hanbing Wang |
| author_sort | Mei Li |
| collection | DOAJ |
| description | IntroductionGoat-derived adeno-associated virus (AAV) vectors, such as AAV Go.1, represent a novel platform for gene therapy due to their unique origin and potential advantages in transduction efficiency and immune evasion. However, their therapeutic potential and biological properties remain underexplored.MethodsIn this study, we developed a recombinant AAV (rAAV) Go.1 by replacing the goat AAV rep gene with the standard AAV2-rep gene to improve packaging efficiency. We compared the transduction efficiency of rAAV Go.1 with that of AAV5, a closely related serotype with 95% genome similarity, both in vitro and in vivo. Additionally, we assessed immune evasion properties by evaluating resistance to neutralization using sera from rAAV5-immunized mice and human volunteers. To further enhance transduction efficiency, we introduced site-specific mutations in the VP1 unique (VP1u) region and VP1/2 common region.ResultsThe rep gene modification led to a significantly higher packaging efficiency for rAAV Go.1 compared to the original goat AAV. rAAV Go.1 exhibited markedly higher transduction efficiency than AAV5 in both in vitro and in vivo models. Furthermore, rAAV Go.1 demonstrated a 4-fold increase in resistance to neutralization by sera from rAAV5-immunized mice. A study involving 20 healthy volunteers revealed that high-titer neutralizing antibodies had a more pronounced inhibitory effect on rAAV5 compared to rAAV Go.1. Mutagenesis studies identified key modifications that enhanced viral properties: K32R, K91R, and K122R mutations in the VP1u region significantly improved viral production, while K137R (VP1u) enhanced transduction efficiency in vitro and in vivo.DiscussionOur findings highlight the potential of rAAV Go.1 as an improved gene therapy vector with superior transduction efficiency and enhanced immune evasion. The identified VP1 mutations further optimize viral properties, making rAAV Go.1 a promising candidate for future therapeutic applications. |
| format | Article |
| id | doaj-art-94a8eab60daf49fda7b8afe0547ec83e |
| institution | DOAJ |
| issn | 2296-858X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Medicine |
| spelling | doaj-art-94a8eab60daf49fda7b8afe0547ec83e2025-08-20T03:07:28ZengFrontiers Media S.A.Frontiers in Medicine2296-858X2025-04-011210.3389/fmed.2025.15544491554449Distinct infectivity and neutralization antibody responses in the highly homologous AAV Go.1 and AAV5Mei Li0Haixiao Ma1Yang Wu2Yunling Gao3Jie Wang4Jie Wang5Hanbing Wang6Department of Anesthesiology, First People Hospital of Foshan, Foshan, ChinaInstitute of Neuroscience and Brain Diseases; Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, ChinaState Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences-Wuhan National Laboratory for Optoelectronics, Wuhan, ChinaInstitute of Neuroscience and Brain Diseases; Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, ChinaInstitute of Neuroscience and Brain Diseases; Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, ChinaDepartment of Radiology, Songjiang Research Institute, Shanghai Key Laboratory of Emotions and Affective Disorders, Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, ChinaDepartment of Anesthesiology, First People Hospital of Foshan, Foshan, ChinaIntroductionGoat-derived adeno-associated virus (AAV) vectors, such as AAV Go.1, represent a novel platform for gene therapy due to their unique origin and potential advantages in transduction efficiency and immune evasion. However, their therapeutic potential and biological properties remain underexplored.MethodsIn this study, we developed a recombinant AAV (rAAV) Go.1 by replacing the goat AAV rep gene with the standard AAV2-rep gene to improve packaging efficiency. We compared the transduction efficiency of rAAV Go.1 with that of AAV5, a closely related serotype with 95% genome similarity, both in vitro and in vivo. Additionally, we assessed immune evasion properties by evaluating resistance to neutralization using sera from rAAV5-immunized mice and human volunteers. To further enhance transduction efficiency, we introduced site-specific mutations in the VP1 unique (VP1u) region and VP1/2 common region.ResultsThe rep gene modification led to a significantly higher packaging efficiency for rAAV Go.1 compared to the original goat AAV. rAAV Go.1 exhibited markedly higher transduction efficiency than AAV5 in both in vitro and in vivo models. Furthermore, rAAV Go.1 demonstrated a 4-fold increase in resistance to neutralization by sera from rAAV5-immunized mice. A study involving 20 healthy volunteers revealed that high-titer neutralizing antibodies had a more pronounced inhibitory effect on rAAV5 compared to rAAV Go.1. Mutagenesis studies identified key modifications that enhanced viral properties: K32R, K91R, and K122R mutations in the VP1u region significantly improved viral production, while K137R (VP1u) enhanced transduction efficiency in vitro and in vivo.DiscussionOur findings highlight the potential of rAAV Go.1 as an improved gene therapy vector with superior transduction efficiency and enhanced immune evasion. The identified VP1 mutations further optimize viral properties, making rAAV Go.1 a promising candidate for future therapeutic applications.https://www.frontiersin.org/articles/10.3389/fmed.2025.1554449/fullgene therapyrAAV Go.1cell transfectionneutralizing antibodypackaging efficiency |
| spellingShingle | Mei Li Haixiao Ma Yang Wu Yunling Gao Jie Wang Jie Wang Hanbing Wang Distinct infectivity and neutralization antibody responses in the highly homologous AAV Go.1 and AAV5 Frontiers in Medicine gene therapy rAAV Go.1 cell transfection neutralizing antibody packaging efficiency |
| title | Distinct infectivity and neutralization antibody responses in the highly homologous AAV Go.1 and AAV5 |
| title_full | Distinct infectivity and neutralization antibody responses in the highly homologous AAV Go.1 and AAV5 |
| title_fullStr | Distinct infectivity and neutralization antibody responses in the highly homologous AAV Go.1 and AAV5 |
| title_full_unstemmed | Distinct infectivity and neutralization antibody responses in the highly homologous AAV Go.1 and AAV5 |
| title_short | Distinct infectivity and neutralization antibody responses in the highly homologous AAV Go.1 and AAV5 |
| title_sort | distinct infectivity and neutralization antibody responses in the highly homologous aav go 1 and aav5 |
| topic | gene therapy rAAV Go.1 cell transfection neutralizing antibody packaging efficiency |
| url | https://www.frontiersin.org/articles/10.3389/fmed.2025.1554449/full |
| work_keys_str_mv | AT meili distinctinfectivityandneutralizationantibodyresponsesinthehighlyhomologousaavgo1andaav5 AT haixiaoma distinctinfectivityandneutralizationantibodyresponsesinthehighlyhomologousaavgo1andaav5 AT yangwu distinctinfectivityandneutralizationantibodyresponsesinthehighlyhomologousaavgo1andaav5 AT yunlinggao distinctinfectivityandneutralizationantibodyresponsesinthehighlyhomologousaavgo1andaav5 AT jiewang distinctinfectivityandneutralizationantibodyresponsesinthehighlyhomologousaavgo1andaav5 AT jiewang distinctinfectivityandneutralizationantibodyresponsesinthehighlyhomologousaavgo1andaav5 AT hanbingwang distinctinfectivityandneutralizationantibodyresponsesinthehighlyhomologousaavgo1andaav5 |