Noninvasive Focal Transgene Delivery with Viral Neuronal Tracers in the Marmoset Monkey

Overview

Journal Cell Rep Methods

Specialty Cell Biology

Date 2024 Feb 15

PMID 38359822

Authors

T Vincenza Parks

Diego Szczupak

Sang-Ho Choi

David J Schaeffer

Affiliations

Soon will be listed here.

Abstract

We establish a reliable method for selectively delivering adeno-associated viral vectors (AAVs) across the blood-brain barrier (BBB) in the marmoset without the need for neurosurgical injection. We focally perturbed the BBB (∼1 × 2 mm) in area 8aD of the frontal cortex in four adult marmoset monkeys using low-intensity transcranial focused ultrasound aided by microbubbles. Within an hour of opening the BBB, either AAV2 or AAV9 was delivered systemically via tail-vein injection. In all four marmosets, fluorescence-encoded neurons were observed at the site of BBB perturbation, with AAV2 showing a sparse distribution of transduced neurons when compared to AAV9. The results are compared to direct intracortical injections of anterograde tracers into area 8aD and similar (albeit sparser) long-range connectivity was observed. With evidence of transduced neurons specific to the region of BBB opening as well as long-distance tracing, we establish a framework for focal noninvasive transgene delivery to the marmoset brain.

Citing Articles

Focused ultrasound widely broadens AAV-delivered Cas9 distribution and activity.

Gumusgoz E, Kasiri S, Youssef I, Verma M, Chopra R, Villarreal Acha D Gene Ther. 2025; .

PMID: 39893321 DOI: 10.1038/s41434-025-00517-w.

Adeno-associated virus vector delivery to the brain: Technology advancements and clinical applications.

Ye D, Chukwu C, Yang Y, Hu Z, Chen H Adv Drug Deliv Rev. 2024; 211:115363.

PMID: 38906479 PMC: 11892011. DOI: 10.1016/j.addr.2024.115363.

Focused ultrasound-mediated delivery of viral neuronal tracers in marmosets.

Owusu-Yaw B, Todd N Cell Rep Methods. 2024; 4(2):100719.

PMID: 38412835 PMC: 10921033. DOI: 10.1016/j.crmeth.2024.100719.

References

Burgess A, Shah K, Hough O, Hynynen K . Focused ultrasound-mediated drug delivery through the blood-brain barrier. Expert Rev Neurother. 2015; 15(5):477-91. PMC: 4702264. DOI: 10.1586/14737175.2015.1028369. View

Murrey D, Naughton B, Duncan F, Meadows A, Ware T, Campbell K . Feasibility and safety of systemic rAAV9-hNAGLU delivery for treating mucopolysaccharidosis IIIB: toxicology, biodistribution, and immunological assessments in primates. Hum Gene Ther Clin Dev. 2014; 25(2):72-84. PMC: 4124586. DOI: 10.1089/humc.2013.208. View

Hori Y, Clery J, Selvanayagam J, Schaeffer D, Johnston K, Menon R . Interspecies activation correlations reveal functional correspondences between marmoset and human brain areas. Proc Natl Acad Sci U S A. 2021; 118(37). PMC: 8449355. DOI: 10.1073/pnas.2110980118. View

Schaeffer D, Gilbert K, Ghahremani M, Gati J, Menon R, Everling S . Intrinsic functional clustering of anterior cingulate cortex in the common marmoset. Neuroimage. 2018; 186:301-307. DOI: 10.1016/j.neuroimage.2018.11.005. View

Lin M, Takahashi Y, Huo B, Hanada M, Nagashima J, Hata J . A high-throughput neurohistological pipeline for brain-wide mesoscale connectivity mapping of the common marmoset. Elife. 2019; 8. PMC: 6384052. DOI: 10.7554/eLife.40042. View

Weber T . Anti-AAV Antibodies in AAV Gene Therapy: Current Challenges and Possible Solutions. Front Immunol. 2021; 12:658399. PMC: 8010240. DOI: 10.3389/fimmu.2021.658399. View

Abe H, Tani T, Mashiko H, Kitamura N, Hayami T, Watanabe S . Axonal Projections From the Middle Temporal Area in the Common Marmoset. Front Neuroanat. 2018; 12:89. PMC: 6218423. DOI: 10.3389/fnana.2018.00089. View

Goertsen D, Flytzanis N, Goeden N, Chuapoco M, Cummins A, Chen Y . AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset. Nat Neurosci. 2021; 25(1):106-115. DOI: 10.1038/s41593-021-00969-4. View

Burman K, Bakola S, Richardson K, Reser D, Rosa M . Patterns of afferent input to the caudal and rostral areas of the dorsal premotor cortex (6DC and 6DR) in the marmoset monkey. J Comp Neurol. 2014; 522(16):3683-716. DOI: 10.1002/cne.23633. View

10.

Alonso A, Reinz E, Leuchs B, Kleinschmidt J, Fatar M, Geers B . Focal Delivery of AAV2/1-transgenes Into the Rat Brain by Localized Ultrasound-induced BBB Opening. Mol Ther Nucleic Acids. 2013; 2:e73. PMC: 3586801. DOI: 10.1038/mtna.2012.64. View

11.

Stavarache M, Petersen N, Jurgens E, Milstein E, Rosenfeld Z, Ballon D . Safe and stable noninvasive focal gene delivery to the mammalian brain following focused ultrasound. J Neurosurg. 2018; 130(3):989-998. DOI: 10.3171/2017.8.JNS17790. View

12.

Kofoed R, Dibia C, Noseworthy K, Xhima K, Vacaresse N, Hynynen K . Efficacy of gene delivery to the brain using AAV and ultrasound depends on serotypes and brain areas. J Control Release. 2022; 351:667-680. DOI: 10.1016/j.jconrel.2022.09.048. View

13.

Majka P, Rosa M, Bai S, Chan J, Huo B, Jermakow N . Unidirectional monosynaptic connections from auditory areas to the primary visual cortex in the marmoset monkey. Brain Struct Funct. 2018; 224(1):111-131. PMC: 6373361. DOI: 10.1007/s00429-018-1764-4. View

14.

Parks T, Szuzupak D, Choi S, Alikaya A, Mou Y, Silva A . Noninvasive disruption of the blood-brain barrier in the marmoset monkey. Commun Biol. 2023; 6(1):806. PMC: 10397190. DOI: 10.1038/s42003-023-05185-3. View

15.

Rosa M, Palmer S, Gamberini M, Burman K, Yu H, Reser D . Connections of the dorsomedial visual area: pathways for early integration of dorsal and ventral streams in extrastriate cortex. J Neurosci. 2009; 29(14):4548-63. PMC: 6665729. DOI: 10.1523/JNEUROSCI.0529-09.2009. View

16.

Abbott C, Percival K, Martin P, Grunert U . Amacrine and bipolar inputs to midget and parasol ganglion cells in marmoset retina. Vis Neurosci. 2012; 29(3):157-68. DOI: 10.1017/S095252381200017X. View

17.

Trinh D, Nash J, Goertz D, Hynynen K, Bulner S, Iqbal U . Microbubble drug conjugate and focused ultrasound blood brain barrier delivery of AAV-2 SIRT-3. Drug Deliv. 2022; 29(1):1176-1183. PMC: 9004516. DOI: 10.1080/10717544.2022.2035855. View

18.

Wang S, Olumolade O, Sun T, Samiotaki G, Konofagou E . Noninvasive, neuron-specific gene therapy can be facilitated by focused ultrasound and recombinant adeno-associated virus. Gene Ther. 2014; 22(1):104-10. PMC: 4294560. DOI: 10.1038/gt.2014.91. View

19.

Bakken T, Jorstad N, Hu Q, Lake B, Tian W, Kalmbach B . Comparative cellular analysis of motor cortex in human, marmoset and mouse. Nature. 2021; 598(7879):111-119. PMC: 8494640. DOI: 10.1038/s41586-021-03465-8. View

20.

Karakatsani M, Wang S, Samiotaki G, Kugelman T, Olumolade O, Acosta C . Amelioration of the nigrostriatal pathway facilitated by ultrasound-mediated neurotrophic delivery in early Parkinson's disease. J Control Release. 2019; 303:289-301. PMC: 6618306. DOI: 10.1016/j.jconrel.2019.03.030. View