TY - JOUR
T1 - The engineered AAV2-HBKO promotes non-invasive gene delivery to large brain regions beyond ultrasound targeted sites
AU - Kofoed, Rikke Hahn
AU - Noseworthy, Kate
AU - Wu, Kathleen
AU - Sivadas, Shuruthisai
AU - Stanek, Lisa
AU - Elmer, Bradford
AU - Hynynen, Kullervo
AU - Shihabuddin, Lamya S.
AU - Aubert, Isabelle
N1 - Funding Information:
The authors thank Kristina Mikloska, Shawna Rideout, and Viva Chan for their expertise on managing the MRIgFUS instruments and preparing the animals during the MRIgFUS procedures. We acknowledge Dr. Nathalie Vacaresse for her inputs during manuscript writing. For imaging, we thank the Centre for Flow Cytometry & Microscopy and facility manager Paul Oleynik at Sunnybrook Research Institute for access to the Zeiss Z1 Axio Observer confocal spinning disk. We are grateful to the Microscopy and Imaging Laboratory and facility manager Dr. Lindsey Fiddes at the University of Toronto for guidance and access to the Zeiss Z1 Axio Observer confocal spinning disk and Zeiss Axio Scan.Z1. We thank Dr. Yutaka Amemiya at the Genomics Core Facility, Sunnybrook Research Institute, for digital droplet PCR analysis, and histotechnologist Petia Stefanova, MSc, at the histology facility, Sunnybrook Research Institute, for cryostat sectioning. Salary support was provided by the Alzheimer Society Research Program (post-doctoral fellowship 19-10 to R.H.K.) and the Carlsberg Internationalisation Fellowship (#CF20-0379 to R.H.K.). This research was undertaken, in part, thanks to funding from the Canada Research Chairs Program ( CRC Tier 1 in Brain Repair and Regeneration to I.A.). This work was funded by Sanofi iAwards, Canadian Institutes of Health Research (to I.A.: 137064 , 166184 , and 168906 and to K.H.: 154272 ), National Institutes of Biomedical Imaging and Bioengineering of the National Institutes of Health (to K.H.: RO1-EB003268. ), and Temerty Chair in Focused Ultrasound Research (to K.H.). Additional funding was received through Sunnybrook Foundation with thanks to the FDC Foundation, WB Family Foundation, Gerald and Carla Connor, andfrom the Canada First Research Excellence Fund, Medicine by Design, University of Toronto Fellowship (to K.N.), Margaret and Howard GAMBLE Research Grant (to K.N.), and Branch Out Neurological Foundation Graduate Research Grant (to K.N.).
Funding Information:
The authors thank Kristina Mikloska, Shawna Rideout, and Viva Chan for their expertise on managing the MRIgFUS instruments and preparing the animals during the MRIgFUS procedures. We acknowledge Dr. Nathalie Vacaresse for her inputs during manuscript writing. For imaging, we thank the Centre for Flow Cytometry & Microscopy and facility manager Paul Oleynik at Sunnybrook Research Institute for access to the Zeiss Z1 Axio Observer confocal spinning disk. We are grateful to the Microscopy and Imaging Laboratory and facility manager Dr. Lindsey Fiddes at the University of Toronto for guidance and access to the Zeiss Z1 Axio Observer confocal spinning disk and Zeiss Axio Scan.Z1. We thank Dr. Yutaka Amemiya at the Genomics Core Facility, Sunnybrook Research Institute, for digital droplet PCR analysis, and histotechnologist Petia Stefanova, MSc, at the histology facility, Sunnybrook Research Institute, for cryostat sectioning. Salary support was provided by the Alzheimer Society Research Program (post-doctoral fellowship 19-10 to R.H.K.) and the Carlsberg Internationalisation Fellowship (#CF20-0379 to R.H.K.). This research was undertaken, in part, thanks to funding from the Canada Research Chairs Program (CRC Tier 1 in Brain Repair and Regeneration to I.A.). This work was funded by Sanofi iAwards, Canadian Institutes of Health Research (to I.A.: 137064, 166184, and 168906 and to K.H.: 154272), National Institutes of Biomedical Imaging and Bioengineering of the National Institutes of Health (to K.H.: RO1-EB003268.), and Temerty Chair in Focused Ultrasound Research (to K.H.). Additional funding was received through Sunnybrook Foundation with thanks to the FDC Foundation, WB Family Foundation, Gerald and Carla Connor, andfrom the Canada First Research Excellence Fund, Medicine by Design, University of Toronto Fellowship (to K.N.), Margaret and Howard GAMBLE Research Grant (to K.N.), and Branch Out Neurological Foundation Graduate Research Grant (to K.N.). R.H.K. L.S. B.E. K.H. L.S.S. and I.A. designed the study. R.H.K. performed the animal experiments. R.H.K. K.N. K.W. and S.S. processed and analyzed the tissue. The manuscript was drafted and figures created by R.H.K. K.N. K.W. and S.S. and edited by I.A. All authors have proof-read and approved the manuscript. L.S. and L.S.S. were paid employees of Sanofi when most of the work was done. B.E. is a paid employee of Sanofi.
Publisher Copyright:
© 2022 The Author(s)
PY - 2022/12/8
Y1 - 2022/12/8
N2 - Magnetic resonance imaging-guided focused ultrasound combined with microbubbles injected in the bloodstream (MRIgFUS) temporarily increases the permeability of the blood-brain barrier (BBB), which facilitates the entry of intravenously administered adeno-associated viruses (AAVs) from the blood to targeted brain areas. To date, the properties of the AAVs used for MRIgFUS delivery resulted in cell transduction limited to MRIgFUS-targeted sites. Considering future clinical applications, strategies are needed to deliver genes to multiple locations and large brain volumes while creating minimal BBB modulation. Here we combine MRIgFUS with a vector that has enhanced biodistribution following brain entry, AAV2-HBKO, to mediate broad gene delivery to targeted brain regions at levels with potential therapeutic relevance. Expression of a reporter gene was achieved in 13% and 21% of all neurons present in the striatum and thalamus, respectively, while targeting only 28% of the brain regions with MRIgFUS. Compared with AAV9, MRIgFUS-mediated delivery of AAV2-HBKO showed greater diffusion in the brain and a higher percentage of the neurons expressing the transgene. MRIgFUS AAV2-HBKO gene delivery to the brain has the potential to reach levels that are functionally and clinically relevant, and this even when using relatively low intravenous AAV dosages, compared with what is currently used in clinical trials.
AB - Magnetic resonance imaging-guided focused ultrasound combined with microbubbles injected in the bloodstream (MRIgFUS) temporarily increases the permeability of the blood-brain barrier (BBB), which facilitates the entry of intravenously administered adeno-associated viruses (AAVs) from the blood to targeted brain areas. To date, the properties of the AAVs used for MRIgFUS delivery resulted in cell transduction limited to MRIgFUS-targeted sites. Considering future clinical applications, strategies are needed to deliver genes to multiple locations and large brain volumes while creating minimal BBB modulation. Here we combine MRIgFUS with a vector that has enhanced biodistribution following brain entry, AAV2-HBKO, to mediate broad gene delivery to targeted brain regions at levels with potential therapeutic relevance. Expression of a reporter gene was achieved in 13% and 21% of all neurons present in the striatum and thalamus, respectively, while targeting only 28% of the brain regions with MRIgFUS. Compared with AAV9, MRIgFUS-mediated delivery of AAV2-HBKO showed greater diffusion in the brain and a higher percentage of the neurons expressing the transgene. MRIgFUS AAV2-HBKO gene delivery to the brain has the potential to reach levels that are functionally and clinically relevant, and this even when using relatively low intravenous AAV dosages, compared with what is currently used in clinical trials.
KW - AAV2-HBKO
KW - adeno-associated virus
KW - blood-brain barrier
KW - focused ultrasound
KW - gene delivery
KW - microbubbles
UR - http://www.scopus.com/inward/record.url?scp=85139322804&partnerID=8YFLogxK
U2 - 10.1016/j.omtm.2022.09.011
DO - 10.1016/j.omtm.2022.09.011
M3 - Journal article
C2 - 36284767
AN - SCOPUS:85139322804
SN - 2329-0501
VL - 27
SP - 167
EP - 184
JO - Molecular Therapy Methods and Clinical Development
JF - Molecular Therapy Methods and Clinical Development
ER -