RNAi and chemogenetic reporter co-regulation in primate striatal interneurons.
Journal
Gene therapy
ISSN: 1476-5462
Titre abrégé: Gene Ther
Pays: England
ID NLM: 9421525
Informations de publication
Date de publication:
02 2022
02 2022
Historique:
received:
22
10
2020
accepted:
22
04
2021
revised:
23
03
2021
pubmed:
21
5
2021
medline:
20
4
2022
entrez:
20
5
2021
Statut:
ppublish
Résumé
Using genetic tools to study the functional roles of molecularly specified neuronal populations in the primate brain is challenging, primarily because of specificity and verification of virus-mediated targeting. Here, we report a lentivirus-based system that helps improve specificity and verification by (a) targeting a selected molecular mechanism, (b) in vivo reporting of expression, and (c) allowing the option to independently silence all regional neural activity. Specifically, we modulate cholinergic signaling of striatal interneurons by shRNAmir and pair it with hM4Di_CFP, a chemogenetic receptor that can function as an in vivo and in situ reporter. Quantitative analyses by visual and deep-learning assisted methods show an inverse linear relation between hM4Di_CFP and ChAT protein expression for several shRNAmir constructs. This approach successfully applies shRNAmir to modulating gene expression in the primate brain and shows that hM4Di_CFP can act as a readout for this modulation.
Identifiants
pubmed: 34012109
doi: 10.1038/s41434-021-00260-y
pii: 10.1038/s41434-021-00260-y
pmc: PMC8856958
doi:
Types de publication
Journal Article
Research Support, N.I.H., Intramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
69-80Subventions
Organisme : Intramural NIH HHS
ID : ZIA MH002619
Pays : United States
Informations de copyright
© 2021. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.
Références
Edry E, Lamprecht R, Wagner S, Rosenblum K. Virally mediated gene manipulation in the adult CNS. Front Mol Neurosci. 2011;4:57. https://doi.org/10.3389/fnmol.2011.00057 .
doi: 10.3389/fnmol.2011.00057
pubmed: 22207836
pmcid: 3245970
El-Shamayleh Y, Horwitz GD. Primate optogenetics: progress and prognosis. Proc Natl Acad Sci USA. 2019. https://doi.org/10.1073/pnas.1902284116 .
doi: 10.1073/pnas.1902284116
pubmed: 31871196
pmcid: 6936537
Eldridge MA, Lerchner W, Saunders RC, Kaneko H, Krausz KW, Gonzalez FJ, et al. Chemogenetic disconnection of monkey orbitofrontal and rhinal cortex reversibly disrupts reward value. Nat Neurosci. 2016;19:37–9. https://doi.org/10.1038/nn.4192 .
doi: 10.1038/nn.4192
pubmed: 26656645
Lerchner W, Corgiat B, Der Minassian V, Saunders RC, Richmond BJ. Injection parameters and virus dependent choice of promoters to improve neuron targeting in the nonhuman primate brain. Gene Ther. 2014;21:233–41. https://doi.org/10.1038/gt.2013.75 .
doi: 10.1038/gt.2013.75
pubmed: 24401836
pmcid: 6563815
Salegio EA, Samaranch L, Kells AP, Forsayeth J, Bankiewicz K. Guided delivery of adeno-associated viral vectors into the primate brain. Adv Drug Deliv Rev. 2012;64:598–604. https://doi.org/10.1016/j.addr.2011.10.005 .
doi: 10.1016/j.addr.2011.10.005
pubmed: 22036906
Martel AC, Elseedy H, Lavigne M, Scapula J, Ghestem A, Kremer EJ, et al. Targeted transgene expression in cholinergic interneurons in the monkey striatum using canine adenovirus serotype 2 vectors. Front Mol Neurosci. 2020;13:76. https://doi.org/10.3389/fnmol.2020.00076 .
doi: 10.3389/fnmol.2020.00076
pubmed: 32499678
pmcid: 7242643
Nair RR, Blankvoort S, Lagartos MJ, Kentros C. Enhancer-driven gene expression (EDGE) enables the generation of viral vectors specific to neuronal subtypes. iScience. 2020;23:100888. https://doi.org/10.1016/j.isci.2020.100888 .
doi: 10.1016/j.isci.2020.100888
pubmed: 32087575
pmcid: 7033522
Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, et al. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Nat Neurosci. 2017;20:1172–9. https://doi.org/10.1038/nn.4593 .
doi: 10.1038/nn.4593
pubmed: 28671695
pmcid: 5529245
Jazayeri M, Lindbloom-Brown Z, Horwitz GD. Saccadic eye movements evoked by optogenetic activation of primate V1. Nat Neurosci. 2012;15:1368–70. https://doi.org/10.1038/nn.3210 .
doi: 10.1038/nn.3210
pubmed: 22941109
pmcid: 3458167
Nagai Y, Kikuchi E, Lerchner W, Inoue KI, Ji B, Eldridge MA, et al. PET imaging-guided chemogenetic silencing reveals a critical role of primate rostromedial caudate in reward evaluation. Nat Commun. 2016;7:13605. https://doi.org/10.1038/ncomms13605 .
doi: 10.1038/ncomms13605
pubmed: 27922009
pmcid: 5150653
Gonzales KK, Pare JF, Wichmann T, Smith Y. GABAergic inputs from direct and indirect striatal projection neurons onto cholinergic interneurons in the primate putamen. J Comp Neurol. 2013;521:2502–22. https://doi.org/10.1002/cne.23295 .
doi: 10.1002/cne.23295
pubmed: 23296794
pmcid: 3983787
Santamaria J, Khalfallah O, Sauty C, Brunet I, Sibieude M, Mallet J, et al. Silencing of choline acetyltransferase expression by lentivirus-mediated RNA interference in cultured cells and in the adult rodent brain. J Neurosci Res. 2009;87:532–44. https://doi.org/10.1002/jnr.21866 .
doi: 10.1002/jnr.21866
pubmed: 18803282
Bonaventura J, Eldridge MAG, Hu F, Gomez JL, Sanchez-Soto M, Abramyan AM, et al. High-potency ligands for DREADD imaging and activation in rodents and monkeys. Nat Commun. 2019;10:4627. https://doi.org/10.1038/s41467-019-12236-z .
doi: 10.1038/s41467-019-12236-z
pubmed: 31604917
pmcid: 6788984
Qian H, Kang X, Hu J, Zhang D, Liang Z, Meng F, et al. Reversing a model of Parkinson’s disease with in situ converted nigral neurons. Nature. 2020;582:550–6. https://doi.org/10.1038/s41586-020-2388-4 .
doi: 10.1038/s41586-020-2388-4
pubmed: 32581380
pmcid: 7521455
Stegmeier F, Hu G, Rickles RJ, Hannon GJ, Elledge SJ. A lentiviral microRNA-based system for single-copy polymerase II-regulated RNA interference in mammalian cells. Proc Natl Acad Sci USA. 2005;102:13212–7. https://doi.org/10.1073/pnas.0506306102 .
doi: 10.1073/pnas.0506306102
pubmed: 16141338
pmcid: 1196357
Halova A, Janoutova J, Ewerlingova L, Janout V, Bonczek O, Zeman T, et al. CHAT gene polymorphism rs3810950 is associated with the risk of Alzheimer’s disease in the Czech population. J Biomed Sci. 2018;25:41. https://doi.org/10.1186/s12929-018-0444-2 .
doi: 10.1186/s12929-018-0444-2
pubmed: 29759072
pmcid: 5950140
Liu YP, Haasnoot J, ter Brake O, Berkhout B, Konstantinova P. Inhibition of HIV-1 by multiple siRNAs expressed from a single microRNA polycistron. Nucleic Acids Res. 2008;36:2811–24. https://doi.org/10.1093/nar/gkn109 .
doi: 10.1093/nar/gkn109
pubmed: 18346971
pmcid: 2396423
Rhesus Monkey Genome Project: https://www.hgsc.bcm.edu/non-human-primates/rhesus-monkey-genome-project .
Falk T, Mai D, Bensch R, Cicek O, Abdulkadir A, Marrakchi Y, et al. U-Net: deep learning for cell counting, detection, and morphometry. Nat Methods. 2019;16:67–70. https://doi.org/10.1038/s41592-018-0261-2 .
doi: 10.1038/s41592-018-0261-2
pubmed: 30559429
Eckenstein F, Sofroniew MV. Identification of central cholinergic neurons containing both choline acetyltransferase and acetylcholinesterase and of central neurons containing only acetylcholinesterase. J Neurosci. 1983;3:2286–91.
doi: 10.1523/JNEUROSCI.03-11-02286.1983
Gonzales KK, Smith Y. Cholinergic interneurons in the dorsal and ventral striatum: anatomical and functional considerations in normal and diseased conditions. Ann NY Acad Sci. 2015;1349:1–45. https://doi.org/10.1111/nyas.12762 .
doi: 10.1111/nyas.12762
pubmed: 25876458
Misgeld T, Burgess RW, Lewis RM, Cunningham JM, Lichtman JW, Sanes JR. Roles of neurotransmitter in synapse formation: development of neuromuscular junctions lacking choline acetyltransferase. Neuron. 2002;36:635–48. https://doi.org/10.1016/s0896-6273(02)01020-6 .
doi: 10.1016/s0896-6273(02)01020-6
pubmed: 12441053
Alexeyev MF, Fayzulin R, Shokolenko IN, Pastukh V. A retro-lentiviral system for doxycycline-inducible gene expression and gene knockdown in cells with limited proliferative capacity. Mol Biol Rep. 2010;37:1987–91. https://doi.org/10.1007/s11033-009-9647-7 .
doi: 10.1007/s11033-009-9647-7
pubmed: 19655272
Xie C, Chen YL, Wang DF, Wang YL, Zhang TP, Li H, et al. SgRNA expression of CRIPSR-Cas9 system based on MiRNA polycistrons as a versatile tool to manipulate multiple and tissue-specific genome editing. Sci Rep. 2017;7:5795. https://doi.org/10.1038/s41598-017-06216-w .
doi: 10.1038/s41598-017-06216-w
pubmed: 28724960
pmcid: 5517485
Fredericks JM, Dash KE, Jaskot EM, Bennett TW, Lerchner W, Dold G, et al. Methods for mechanical delivery of viral vectors into rhesus monkey brain. J Neurosci Methods. 2020;339:108730. https://doi.org/10.1016/j.jneumeth.2020.108730 .
doi: 10.1016/j.jneumeth.2020.108730
pubmed: 32302596
pmcid: 7238764
Saunders RC, Aigner TG, Frank JA. Magnetic-resonance-imaging of the rhesus-monkey brain—use for stereotaxic neurosurgery. Exp Brain Res. 1990;81:443–6.
doi: 10.1007/BF00228139