Third-generation rabies viral vectors allow nontoxic retrograde targeting of projection neurons with greatly increased efficiency

Jin, Lei; Sullivan, Heather A.; Zhu, Mulangma; Lea, Nicholas E.; Lavin, Thomas K.; Fu, Xin; Matsuyama, Makoto; Hou, YuanYuan; Feng, Guoping; Wickersham, Ian R.

doi:10.1016/j.crmeth.2023.100644

Cited by 3 publications

(4 citation statements)

References 68 publications

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“…Data distribution was assumed to be normal, but this was not formally tested. No statistical methods were used to predetermine sample sizes, but our sample sizes are similar to those reported in previous publications 50 and are as large as practical given the large number of conditions tested.…”

Section: Methodsmentioning

confidence: 71%

“…Perhaps notably, our finding that ∆GL RVs do not spread when not complemented by L expression in trans , even when G is provided in trans , indicates that deletion of either L or G alone is sufficient to prevent transsynaptic spread. This suggests the possibility of a third-generation monosynaptic tracing system, based on single-deletion-mutant ∆L viruses 50 complemented by expression in trans of just L.…”

Section: Discussionmentioning

confidence: 99%

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Long-term labeling and imaging of synaptically connected neuronal networks in vivo using double-deletion-mutant rabies viruses

Jin,

Sullivan,

Zhu

et al. 2024

Nat Neurosci

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Rabies-virus-based monosynaptic tracing is a widely used technique for mapping neural circuitry, but its cytotoxicity has confined it primarily to anatomical applications. Here we present a second-generation system for labeling direct inputs to targeted neuronal populations with minimal toxicity, using double-deletion-mutant rabies viruses. Viral spread requires expression of both deleted viral genes in trans in postsynaptic source cells. Suppressing this expression with doxycycline following an initial period of viral replication reduces toxicity to postsynaptic cells. Longitudinal two-photon imaging in vivo indicated that over 90% of both presynaptic and source cells survived for the full 12-week course of imaging. Ex vivo whole-cell recordings at 5 weeks postinfection showed that the second-generation system perturbs input and source cells much less than the first-generation system. Finally, two-photon calcium imaging of labeled networks of visual cortex neurons showed that their visual response properties appeared normal for 10 weeks, the longest we followed them.

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Section: Methodsmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 99%

Long-term labeling and imaging of synaptically connected neuronal networks in vivo using double-deletion-mutant rabies viruses

Jin,

Sullivan,

Zhu

et al. 2024

Nat Neurosci

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“…One limitation of the experimental platform is the neurotoxic potential associated with the use of rabies virus for retrograde tracing over time. [91][92][93] This issue underscores the importance of utilizing and further adapting less toxic rabies-based labeling strategies for glioblastoma, 93,94 to enable longer observation periods without adverse effects on neuronal health. Further, rabies virus-mediated retrograde tracing did not label all synaptic inputs in previous work, illustrating that the labeled connected TUM are possibly still an underestimation of the entire neuronal connectome of glioblastoma.…”

Section: Limitations Of the Studymentioning

confidence: 99%

Characterizing and targeting glioblastoma neuron-tumor networks with retrograde tracing

Tetzlaff,

Reyhan,

Bengtson

et al. 2024

Preprint

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Glioblastomas are heterogeneous brain tumors, notorious for their invasive behavior and resistance to therapy. Neuron-to-glioma synapses have been identified to promote glioblastoma invasion and proliferation. However, a comprehensive characterization of tumor-connected neurons has been hampered by a lack of technologies. Here, we adapted retrograde tracing with a modified rabies virus system to characterize and manipulate connected neuron-tumor networks. Glioblastoma rapidly integrated into neural circuits across the brain engaging in widespread functional communication, with acetylcholinergic and glutamatergic neurons driving glioblastoma progression. We uncovered patient-specific and tumor cell state-dependent differences in synaptogenic gene expression driving neuron-tumor connectivity and subsequent invasivity. Importantly, radiotherapy enhanced neuron-tumor connectivity by increased neuronal activity. In turn, simultaneous inhibition of AMPA receptors and radiotherapy showed increased therapeutic effects, indicative of a role for neuron-to-glioma synapses in contributing to therapeutic resistance. Lastly, rabies-mediated genetic ablation of tumor-connected neurons halted glioblastoma progression, offering a novel viral strategy to target glioblastoma. Together, this study provides a comprehensive framework for basic research and clinical translation of synaptic neuron-cancer interactions to target glioblastoma.

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“…In this issue of Cell Reports Methods , Jin and colleagues describe the synthesis and validation of a “third generation” B19-based rabies viral vector called RVΔL ( Figure 1 ). 11 RVΔL builds on the best features of its predecessors, specifically low toxicity (like RVΔGL) and high viral titer (like RVΔG). Specifically, the authors show that when only L is deleted from the rabies genome, it gains a major growth advantage over RVΔGL when propagated on complementing cells in vitro, reaching a viral titer similar to first generation RVΔG rabies.…”

Section: Main Textmentioning

confidence: 99%

Third-generation rabies viral vectors have low toxicity and improved efficiency as retrograde labeling tools

Schwarz

2023

Cell Reports Methods

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Third-generation rabies viral vectors allow nontoxic retrograde targeting of projection neurons with greatly increased efficiency

Cited by 3 publications

References 68 publications

Long-term labeling and imaging of synaptically connected neuronal networks in vivo using double-deletion-mutant rabies viruses

Long-term labeling and imaging of synaptically connected neuronal networks in vivo using double-deletion-mutant rabies viruses

Characterizing and targeting glioblastoma neuron-tumor networks with retrograde tracing

Third-generation rabies viral vectors have low toxicity and improved efficiency as retrograde labeling tools

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