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Cited by 2Virus-based gene therapy researchRNA Interference and Gene DeliveryBarrier Structure and Function Studies

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Bridge protein-mediated viral targeting of cells expressing endogenous μ-opioid G protein-coupled receptors in the mouse and monkey brain

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Kamaguchi, Riki, Amemori, Satoko, Amemori, Ken-ichi, Osakada, Fumitaka·2025·Neuroscience Research
DOI 10.1016/j.neures.2025.02.007

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Abstract

Targeting specific cell types is essential for understanding their functional roles in the brain. Although genetic approaches enable cell-type-specific targeting in animals, their application to higher mammalian species, such as nonhuman primates, remains challenging. Here, we developed a nontransgenic method using bridge proteins to direct viral vectors to cells endogenously expressing μ-opioid receptors (MORs), a G protein-coupled receptor. The bridge protein comprises the avian viral receptor TVB, the MOR ligand β-endorphin (βed), and an interdomain linker. EnvB-enveloped viruses bind to the TVB component, followed by the interaction of βed with MORs, triggering viral infection in MOR-expressing cells. We optimized the secretion signals, domain arrangements, and interdomain linkers of the bridge proteins to maximize viral targeting efficiency and specificity. Alternative configurations incorporating different ligands and viral receptors also induced viral infection in MOR-expressing cells. The optimized βed-f2-TVB bridge protein with EnvB-pseudotyped lentiviruses induced infection in MOR-expressing cells in the striatum of mice and monkeys. An intersectional approach combining βed-f2-TVB with a neuron-specific promoter refined cell-type specificity. This study establishes the foundation for the rational bridge protein design and the feasibility of targeting G protein-coupled receptors beyond tyrosine kinase receptors, thereby expanding targetable cell types in the brain and throughout the body.

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Research works printed but not linked: 84

  1. Adamczyk, 2023, Glial-neuronal interactions in neurological disorders: Molecular mechanisms and potential points for intervention, Int. J. Mol. Sci., 24, 6274, 10.3390/ijms24076274 · DOI
  2. Alfthan, 1995, Properties of a single-chain antibody containing different linker peptides, Protein Eng. Des. Sel., 8, 725, 10.1093/protein/8.7.725 · DOI
  3. Amiche, 1990, Characterisation and visualisation of [3H]dermorphin binding to mu opioid receptors in the rat brain. Combined high selectivity and affinity in a natural peptide agonist for the morphine (mu) receptor, Eur. J. Biochem., 189, 625, 10.1111/j.1432-1033.1990.tb15531.x · DOI
  4. Arai, 2001, Design of the linkers which effectively separate domains of a bifunctional fusion protein, Protein Eng., 14, 529, 10.1093/protein/14.8.529 · DOI
  5. Argos, 1990, An investigation of oligopeptides linking domains in protein tertiary structures and possible candidates for general gene fusion, J. Mol. Biol., 211, 943, 10.1016/0022-2836(90)90085-Z · DOI
  6. Arttamangkul, 2019, Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry, ELife, 8, 10.7554/eLife.49319 · DOI
  7. Atasoy, 2008, A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping, J. Neurosci., 28, 7025, 10.1523/JNEUROSCI.1954-08.2008 · DOI
  8. Bai, 2006, Improving the oral efficacy of recombinant granulocyte colony-stimulating factor and transferrin fusion protein by spacer optimization, Pharm. Res., 23, 2116, 10.1007/s11095-006-9059-5 · DOI
  9. Balliet, 1999, Production and characterization of a soluble, active form of Tva, the subgroup A avian sarcoma and leukosis virus receptor, J. Virol., 73, 3054, 10.1128/JVI.73.4.3054-3061.1999 · DOI
  10. Barnard, 2006, Avian sarcoma and leukosis virus-receptor interactions: from classical genetics to novel insights into virus-cell membrane fusion, Virology, 344, 25, 10.1016/j.virol.2005.09.021 · DOI
  11. Barnard, 2003, Alpharetrovirus envelope-receptor interactions, Curr. Top. Microbiol. Immunol., 281, 107
  12. Bian, 2005, Selective gene transfer to tumor cells by recombinant Newcastle Disease Virus via a bispecific fusion protein, Int. J. Oncol., 26, 431
  13. Boerger, 1999, Retroviral vectors preloaded with a viral receptor-ligand bridge protein are targeted to specific cell types, Proc. Natl. Acad. Sci. U. S. A., 96, 9867, 10.1073/pnas.96.17.9867 · DOI
  14. Branda, 2004, Talking about a revolution: the impact of site-specific recombinases on genetic analyses in mice, Dev. Cell, 6, 7, 10.1016/S1534-5807(03)00399-X · DOI
  15. Brimblecombe, 2017, The striosome and matrix compartments of the striatum: a path through the labyrinth from neurochemistry toward function, ACS Chem. Neurosci., 8, 235, 10.1021/acschemneuro.6b00333 · DOI
  16. Brockmeier, 2006, Systematic screening of all signal peptides from Bacillus subtilis: a powerful strategy in optimizing heterologous protein secretion in Gram-positive bacteria, J. Mol. Biol., 362, 393, 10.1016/j.jmb.2006.07.034 · DOI
  17. Chen, 2022, Engineered AAVs for non-invasive gene delivery to rodent and non-human primate nervous systems, Neuron, 110, 2242, 10.1016/j.neuron.2022.05.003 · DOI
  18. Chen, 2013, Fusion protein linkers: property, design and functionality, Adv. Drug Deliv. Rev., 65, 1357, 10.1016/j.addr.2012.09.039 · DOI
  19. Cheng, 1973, Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction, Biochem. Pharmacol., 22, 3099, 10.1016/0006-2952(73)90196-2 · DOI · Paper page
  20. Choi, 2011, Monosynaptic inputs to ErbB4-expressing inhibitory neurons in mouse primary somatosensory cortex, J. Comp. Neurol., 519, 3402, 10.1002/cne.22680 · DOI · Paper page
  21. Choi, 2010, Selective viral vector transduction of ErbB4 expressing cortical interneurons in vivo with a viral receptor-ligand bridge protein, Proc. Natl. Acad. Sci. U.S.A., 107, 16703, 10.1073/pnas.1006233107 · DOI
  22. Chuapoco, 2023, Adeno-associated viral vectors for functional intravenous gene transfer throughout the non-human primate brain, Nat. Nanotechnol., 18, 1241, 10.1038/s41565-023-01419-x · DOI
  23. Delfs, 1994, Expression of mu opioid receptor mRNA in rat brain: an in situ hybridization study at the single cell level, J. Comp. Neurol., 345, 46, 10.1002/cne.903450104 · DOI
  24. Ding, 2005, Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice, Cell, 122, 473, 10.1016/j.cell.2005.07.013 · DOI · Paper page
  25. Dobson, 2022, Antigen identification and high-throughput interaction mapping by reprogramming viral entry, Nat. Methods, 19, 449, 10.1038/s41592-022-01436-z · DOI
  26. Erbs, 2015, A mu–delta opioid receptor brain atlas reveals neuronal co-occurrence in subcortical networks, Brain Struct. Funct., 220, 677, 10.1007/s00429-014-0717-9 · DOI
  27. Fenno, 2014, Targeting cells with single vectors using multiple-feature Boolean logic, Nat. Methods, 11, 763, 10.1038/nmeth.2996 · DOI · Paper page
  28. Fenno, 2020, Comprehensive dual- and triple-feature intersectional single-vector delivery of diverse functional payloads to cells of behaving mammals, Neuron, 107, 836, 10.1016/j.neuron.2020.06.003 · DOI
  29. Freudl, 2018, Signal peptides for recombinant protein secretion in bacterial expression systems, Microb. Cell Fact., 17, 52, 10.1186/s12934-018-0901-3 · DOI · Paper page
  30. Gaudriault, 1997, Receptor-induced Internalization of Selective Peptidic μ and Δ Opioid Ligands, J. Biol. Chem., 272, 2880, 10.1074/jbc.272.5.2880 · DOI
  31. Gokce, 2016, Cellular taxonomy of the mouse striatum as revealed by single-cell RNA-seq, Cell Rep., 16, 1126, 10.1016/j.celrep.2016.06.059 · DOI · Paper page
  32. Graybiel, 2023, Striosomes and matrisomes: scaffolds for dynamic coupling of volition and action, Annu. Rev. Neurosci., 46, 359, 10.1146/annurev-neuro-121522-025740 · DOI
  33. Graybuck, 2021, Enhancer viruses for combinatorial cell-subclass-specific labeling, Neuron, 109, 1449, 10.1016/j.neuron.2021.03.011 · DOI · Paper page
  34. Han, 2017, Characterization of tree shrew (Tupaia belangeri) interleukin-6 and its expression pattern in response to exogenous challenge, Int. J. Mol. Med., 40, 1679
  35. He, 2021, Transcriptional and anatomical diversity of medium spiny neurons in the primate striatum, Curr. Biol., 31, 5473, 10.1016/j.cub.2021.10.015 · DOI
  36. Hioki, 2009, High-level transgene expression in neurons by lentivirus with Tet-Off system, Neurosci. Res., 63, 149, 10.1016/j.neures.2008.10.010 · DOI
  37. Horii, 2017, Efficient generation of conditional knockout mice via sequential introduction of lox sites, Sci. Rep., 7, 7891, 10.1038/s41598-017-08496-8 · DOI
  38. Hosohata, 1998, Endomorphin-1 and endomorphin-2 are partial agonists at the human mu-opioid receptor, Eur. J. Pharmacol., 346, 111, 10.1016/S0014-2999(98)00117-4 · DOI
  39. Huang, 2013, Genetic approaches to neural circuits in the mouse, Annu. Rev. Neurosci., 36, 183, 10.1146/annurev-neuro-062012-170307 · DOI · Paper page
  40. Hughes, 2024, A single-vector intersectional AAV strategy for interrogating cellular diversity and brain function, Nat. Neurosci., 27, 1400, 10.1038/s41593-024-01659-7 · DOI
  41. Johnston, 1990, Mechanisms of striatal pattern formation: conservation of mammalian compartmentalization, Brain Res. Dev. Brain Res., 57, 93, 10.1016/0165-3806(90)90189-6 · DOI
  42. Jüttner, 2019, Targeting neuronal and glial cell types with synthetic promoter AAVs in mice, non-human primates and humans, Nat. Neurosci., 22, 1345, 10.1038/s41593-019-0431-2 · DOI · Paper page
  43. Kaseniit, 2023, Modular, programmable RNA sensing using ADAR editing in living cells, Nat. Biotechnol., 41, 482, 10.1038/s41587-022-01493-x · DOI
  44. Kishi, 2014, Common marmoset as a new model animal for neuroscience research and genome editing technology, Dev. Growth Differ., 56, 53, 10.1111/dgd.12109 · DOI
  45. Kodera, 2023, Modeling the marmoset brain using embryonic stem cell-derived cerebral assembloids, Biochem. Biophys. Res. Commun., 657, 119, 10.1016/j.bbrc.2023.03.019 · DOI
  46. Kreitzer, 2008, Striatal plasticity and basal ganglia circuit function, Neuron, 60, 543, 10.1016/j.neuron.2008.11.005 · DOI
  47. Luo, 2018, Genetic dissection of neural circuits: a decade of progress, Neuron, 98, 865, 10.1016/j.neuron.2018.05.004 · DOI · Paper page
  48. Maduna, 2018, Microglia express Mu opioid receptor: insights from transcriptomics and fluorescent reporter mice, Front. Psychiatry, 9, 726, 10.3389/fpsyt.2018.00726 · DOI
  49. Mansour, 1994, Mu, delta, and kappa opioid receptor mRNA expression in the rat CNS: an in situ hybridization study, J. Comp. Neurol., 350, 412, 10.1002/cne.903500307 · DOI
  50. Masaki, 2022, Monosynaptic rabies virus tracing from projection-targeted single neurons, Neurosci. Res., 178, 20, 10.1016/j.neures.2022.01.007 · DOI
  51. Matsui, 1985, Solubilization and characterization of rat brain alpha 2-adrenergic receptor, J. Neurochem., 44, 1625, 10.1111/j.1471-4159.1985.tb08805.x · DOI
  52. Matsushima, 2023, Transcriptional vulnerabilities of striatal neurons in human and rodent models of Huntington’s disease, Nat. Commun., 14, 1, 10.1038/s41467-022-35752-x · DOI
  53. Matsuyama, 2015, Avian sarcoma leukosis virus receptor-envelope system for simultaneous dissection of multiple neural circuits in mammalian brain, Proc. Natl. Acad. Sci. U.S.A., 112, E2947, 10.1073/pnas.1423963112 · DOI
  54. McConalogue, 1999, Activation and internalization of the mu-opioid receptor by the newly discovered endogenous agonists, endomorphin-1 and endomorphin-2, Neuroscience, 90, 1051, 10.1016/S0306-4522(98)00514-4 · DOI
  55. Minami, 1995, Molecular biology of the opioid receptors: structures, functions and distributions, Neurosci. Res., 23, 121, 10.1016/0168-0102(95)00933-K · DOI
  56. Mores, 2019, Arrestin recruitment and signaling by G protein-coupled receptor heteromers, Neuropharmacology, 152, 15, 10.1016/j.neuropharm.2018.11.010 · DOI
  57. Mothes, 2000, Retroviral entry mediated by receptor priming and low pH triggering of an envelope glycoprotein, Cell, 103, 679, 10.1016/S0092-8674(00)00170-7 · DOI · Paper page
  58. Nagai, 2021, Behaviorally consequential astrocytic regulation of neural circuits, Neuron, 109, 576, 10.1016/j.neuron.2020.12.008 · DOI
  59. Nambu, 2023, Dynamic activity model of movement disorders: the fundamental role of the hyperdirect pathway, Mov. Disord., 38, 2145, 10.1002/mds.29646 · DOI
  60. Nectow, 2020, Viral tools for neuroscience, Nat. Rev. Neurosci., 21, 669, 10.1038/s41583-020-00382-z · DOI · Paper page
  61. Neda, 1991, Chemical modification of an ecotropic murine leukemia virus results in redirection of its target cell specificity, J. Biol. Chem., 266, 14143, 10.1016/S0021-9258(18)98654-0 · DOI
  62. Ohtsuka, 2018, i-GONAD: a robust method for in situ germline genome engineering using CRISPR nucleases, Genome Biol., 19, 25, 10.1186/s13059-018-1400-x · DOI
  63. Okigawa, 2021, Cell type- and layer-specific convergence in core and shell neurons of the dorsal lateral geniculate nucleus, J. Comp. Neurol., 529, 2099, 10.1002/cne.25075 · DOI
  64. Osakada, 2013, Design and generation of recombinant rabies virus vectors, Nat. Protoc., 8, 1583, 10.1038/nprot.2013.094 · DOI
  65. Osakada, 2011, New rabies virus variants for monitoring and manipulating activity and gene expression in defined neural circuits, Neuron, 71, 617, 10.1016/j.neuron.2011.07.005 · DOI
  66. Pan, 1987, Structural characterization of human interferon gamma. Heterogeneity of the carboxyl terminus, Eur. J. Biochem., 166, 145, 10.1111/j.1432-1033.1987.tb13494.x · DOI
  67. Paolicelli, 2022, Microglia states and nomenclature: a field at its crossroads, Neuron, 110, 3458, 10.1016/j.neuron.2022.10.020 · DOI · Paper page
  68. Park, 2019, Generation of genetically engineered non-human primate models of brain function and neurological disorders, Am. J. Primatol., 81, 10.1002/ajp.22931 · DOI
  69. Qian, 2022, Programmable RNA sensing for cell monitoring and manipulation, Nature, 610, 713, 10.1038/s41586-022-05280-1 · DOI
  70. Reinisová, 2008, A single-amino-acid substitution in the TvbS1 receptor results in decreased susceptibility to infection by avian sarcoma and leukosis virus subgroups B and D and resistance to infection by subgroup E in vitro and in vivo, J. Virol., 82, 2097, 10.1128/JVI.02206-07 · DOI
  71. Rinderknecht, 1984, Natural human interferon-gamma. Complete amino acid sequence and determination of sites of glycosylation, J. Biol. Chem., 259, 6790, 10.1016/S0021-9258(17)39797-1 · DOI
  72. Romero, 1999, Opioid peptide receptor studies. 12. Buprenorphine is a potent and selective mu/kappa antagonist in the [35S]-GTP-gamma-S functional binding assay, Synapse, 34, 83, 10.1002/(SICI)1098-2396(199911)34:2<83::AID-SYN1>3.0.CO;2-S · DOI
  73. Salimando, 2023, Human OPRM1 and murine Oprm1 promoter driven viral constructs for genetic access to μ-opioidergic cell types, Nat. Commun., 14, 5632, 10.1038/s41467-023-41407-2 · DOI
  74. Shinmyo, 2017, Folding of the cerebral cortex requires Cdk5 in upper-layer neurons in gyrencephalic mammals, Cell Rep., 20, 2131, 10.1016/j.celrep.2017.08.024 · DOI
  75. Smith, 2004, The mature avian leukosis virus subgroup A envelope glycoprotein is metastable, and refolding induced by the synergistic effects of receptor binding and low pH is coupled to infection, J. Virol., 78, 1403, 10.1128/JVI.78.3.1403-1410.2004 · DOI
  76. Snitkovsky, 2001, Targeting avian leukosis virus subgroup a vectors by using a TVA-VEGF bridge protein, J. Virol., 75, 1571, 10.1128/JVI.75.3.1571-1575.2001 · DOI
  77. Sofroniew, 2020, Astrocyte reactivity: subtypes, states, and functions in CNS innate immunity, Trends Immunol., 41, 758, 10.1016/j.it.2020.07.004 · DOI · Paper page
  78. Strebinger, 2023, Cell type-specific delivery by modular envelope design, Nat. Commun., 14, 5141, 10.1038/s41467-023-40788-8 · DOI
  79. Suzuki, 2020, Multiplex Neural Circuit Tracing With G-Deleted Rabies Viral Vectors, Front. Neural Circuits, 13, 1, 10.3389/fncir.2019.00077 · DOI
  80. Teufel, 2022, SignalP 6.0 predicts all five types of signal peptides using protein language models, Nat. Biotechnol., 40, 1023, 10.1038/s41587-021-01156-3 · DOI
  81. Trinh, 2004, Optimization of codon pair use within the (GGGGS)3 linker sequence results in enhanced protein expression, Mol. Immunol., 40, 717, 10.1016/j.molimm.2003.08.006 · DOI
  82. Watakabe, 2012, Visualization of cortical projection neurons with retrograde TET-off lentiviral vector, PLoS One, 7, 10.1371/journal.pone.0046157 · DOI
  83. Yoshizawa, 2018, Reward-predictive neural activities in striatal striosome compartments, eNeuro, 5, 10.1523/ENEURO.0367-17.2018 · DOI
  84. Yu, 2022, Engineered cell entry links receptor biology with single-cell genomics, Cell, 185, 4904, 10.1016/j.cell.2022.11.016 · DOI

Metadata

Authors
Kamaguchi, R., Amemori, S., Amemori, K.-I., & Osakada, F.
Affiliations
  • Nagoya University (JP)
  • Japan Society for the Promotion of Science (JP)
  • Kyoto University (JP)
  • Japan Science and Technology Agency (JP)
Corresponding author
Osakada, Fumitaka
ORCID
Venue
Neuroscience Research
Publisher
Elsevier BV
Published
2025-04
Volume
213
Pages
35-50
ISSN
0168-0102 (ISSN-L)
DOI
10.1016/j.neures.2025.02.007
OpenAlex
W4407438968
Cited by
2
References
7
Citations by year
2026: 2
FWCI
1.18
Type
journal article
Language
en
Open access
Licence
Locations
  • Neuroscience Research (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • Japan Agency for Medical Research and Development (JP24wm0625110h)
  • Japan Science and Technology Agency (JPMJCR1851, JPMJPR14F6)
  • Japan Society for the Promotion of Science (21K07259, 21H05168, 20K21476, 18H02706, 21J40030, 20H03555)
  • Naito Foundation
  • Suzuken Memorial Foundation
  • Takeda Science Foundation
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
Keywords
viral vector targeting, mu-opioid receptor, G protein-coupled receptors, beta-endorphin, striatum, non-human primates
MeSH terms
Animals, Brain, Genetic Vectors, Humans, Mice, Inbred C57BL, Neurons, Lentivirus, Receptors, Opioid, mu, Mice, HEK293 Cells

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CoChat. (2026). Citation check: Bridge protein-mediated viral targeting of cells expressing endogenous μ-opioid G protein-coupled receptors in the mouse and monkey brain. https://app.cochat.ai/papers/10.1016/j.neures.2025.02.007
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Kamaguchi, R., Amemori, S., Amemori, K.-I., & Osakada, F. (2025). Bridge protein-mediated viral targeting of cells expressing endogenous μ-opioid G protein-coupled receptors in the mouse and monkey brain. Neuroscience Research. https://doi.org/10.1016/j.neures.2025.02.007