Intracellular accumulation of AAV6 genomes governs microglia reactivity and the inflammatory response in retinal gene therapy
- 1Nantes University, CHU de Nantes, INSERM UMR 1089, TaRGeT Translational Research in Gene Therapy, F 44200 Nantes, France
Abstract
Adeno-associated viral (AAV) vectors possess immense potential for gene therapy applications. However, microglia can mount an immune response against AAV vectors. Yet, microglia are therapeutic targets since they play a driving role in the progression of neurodegenerative diseases. Thus, it is critical to understand the cellular processes driving the microglia-mediated immune response against AAV vectors. To this end, we modelled AAV6 induced inflammation upon retinal gene therapy in minipigs. Upon AAV6 administration, the IBA1+ cells demonstrated a morphological transition to the amoeboid form, indicating microglial reactivity. We also observed the invasion of peripheral immune cells in the retina. To elucidate the underlying molecular mechanisms, we transduced human iPSC derived microglia-like cells (iMGLs) in vitro with AAV6 and AAV8 respectively. Only AAV6 transduced iMGLs exhibited morphological features and a transcriptomic signature of microglial reactivity. The magnitude of intracellular AAV6 viral genomes was significantly higher as compared to the AAV8 condition. The elevated levels of intracellular AAV6 genomes induced a DNA damage response in AAV6-transduced iMGLs, which is a key feature of neuroinflammation-linked microglial reactivity. Thus, our work highlights the impact of intracellular viral genome accumulation on microglial reactivity and its implications for retinal gene therapy associated inflammation.
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BibTeX
@misc{pereira2026intracellular,
title = {{Intracellular accumulation of AAV6 genomes governs microglia reactivity and the inflammatory response in retinal gene therapy}},
author = {A. Pereira and J.-B. Ducloyer and J. Demilly and E. Toublanc and L. Libeau and B. Robert and A. Mellet and D. Augereau and L. Comtesse and T. Larcher and A. Lebreton and P. Socheleau and M. Grard and M. Journou and C. Couzinie and V. Blouin and M. Weber and G. Le Meur and C. Le Guiner and O. Adjali and T. Cronin},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.09.757886},
url = {https://www.biorxiv.org/content/10.64898/2026.10.09.757886v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Intracellular accumulation of AAV6 genomes governs microglia reactivity and the inflammatory response in retinal gene therapy
AU - A. Pereira
AU - J.-B. Ducloyer
AU - J. Demilly
AU - E. Toublanc
AU - L. Libeau
AU - B. Robert
AU - A. Mellet
AU - D. Augereau
AU - L. Comtesse
AU - T. Larcher
AU - A. Lebreton
AU - P. Socheleau
AU - M. Grard
AU - M. Journou
AU - C. Couzinie
AU - V. Blouin
AU - M. Weber
AU - G. Le Meur
AU - C. Le Guiner
AU - O. Adjali
AU - T. Cronin
PY - 2026
DA - 2026/10/09
PB - bioRxiv
DO - 10.64898/2026.10.09.757886
UR - https://www.biorxiv.org/content/10.64898/2026.10.09.757886v1
AB - Adeno-associated viral (AAV) vectors possess immense potential for gene therapy applications. However, microglia can mount an immune response against AAV vectors. Yet, microglia are therapeutic targets since they play a driving role in the progression of neurodegenerative diseases. Thus, it is critical to understand the cellular processes driving the microglia-mediated immune response against AAV vectors. To this end, we modelled AAV6 induced inflammation upon retinal gene therapy in minipigs. Upon AAV6 administration, the IBA1+ cells demonstrated a morphological transition to the amoeboid form, indicating microglial reactivity. We also observed the invasion of peripheral immune cells in the retina. To elucidate the underlying molecular mechanisms, we transduced human iPSC derived microglia-like cells (iMGLs) in vitro with AAV6 and AAV8 respectively. Only AAV6 transduced iMGLs exhibited morphological features and a transcriptomic signature of microglial reactivity. The magnitude of intracellular AAV6 viral genomes was significantly higher as compared to the AAV8 condition. The elevated levels of intracellular AAV6 genomes induced a DNA damage response in AAV6-transduced iMGLs, which is a key feature of neuroinflammation-linked microglial reactivity. Thus, our work highlights the impact of intracellular viral genome accumulation on microglial reactivity and its implications for retinal gene therapy associated inflammation.
ER -