NIDA IRP Technology Development Initiative Featured Paper

A figure from this study. Copyright: Nature

DART.2: bidirectional synaptic pharmacology with thousandfold cellular specificity

Authors

Brenda C. Shields, Haidun Yan, Shaun S. X. Lim, Sasha C. V. Burwell, Celine M. Cammarata, Elizabeth A. Fleming, S. Aryana Yousefzadeh, Victoria Z. Goldenshtein, Elizabeth W. Kahuno, Purav P. Vagadia, Marie H. Loughran, Lei Zhiquan, Mark E. McDonnell, Miranda L. Scalabrino, Mishek Thapa, Tammy M. Hawley, Greg D. Field, Court Hull, Gary E. Schiltz, Lindsey L. Glickfeld, Allen B. Reitz & Michael R. Tadross

Paper presented by Dr. Rajtarun Madangopal and selected by the NIDA TDI Paper of the Month Committee.

Publication Brief Description

DART.2 advances the original DART.1 technology, which enabled cell-specific drug targeting by tethering drugs to the HaloTag protein, to allow precise investigation of how specific proteins in defined cell types and circuits drive behavior. With up to 3,000-fold enhanced targeting precision, DART.2 allows selective delivery of even potent drugs, such as GABA antagonists, to specific neurons while minimizing off-target effects. It is compatible with brain-wide drug delivery via the lateral ventricle, removing the need for localized infusions, and incorporates fluorescent tracers for real-time visualization and quantification of drug capture, ensuring precise dose control. Uniquely, DART.2 introduces bidirectional modulation, allowing activation or inhibition of neurotransmitter pathways within the same cell populations. This expanded DART.2 toolbox enables intersectional precision—targeting genetically defined postsynaptic cells alongside neurotransmitter-defined presynaptic partners. Future refinements will aim to address challenges in systemic delivery across the blood-brain barrier, improve reversibility, and broaden compatibility with various receptor types, furthering potential for finely tuned studies of the neural dynamics underlying behavior.

Shields, Brenda C; Yan, Haidun; Lim, Shaun S X; Burwell, Sasha C V; Cammarata, Celine M; Fleming, Elizabeth A; Yousefzadeh, S Aryana; Goldenshtein, Victoria Z; Kahuno, Elizabeth W; Vagadia, Purav P; Loughran, Marie H; Zhiquan, Lei; McDonnell, Mark E; Scalabrino, Miranda L; Thapa, Mishek; Hawley, Tammy M; Field, Greg D; Hull, Court; Schiltz, Gary E; Glickfeld, Lindsey L; Reitz, Allen B; Tadross, Michael R

DART.2: bidirectional synaptic pharmacology with thousandfold cellular specificity Journal Article

In: Nat Methods, vol. 21, no. 7, pp. 1288–1297, 2024, ISSN: 1548-7105.

Abstract | Links

@article{pmid38877316,

title = {DART.2: bidirectional synaptic pharmacology with thousandfold cellular specificity},

author = {Brenda C Shields and Haidun Yan and Shaun S X Lim and Sasha C V Burwell and Celine M Cammarata and Elizabeth A Fleming and S Aryana Yousefzadeh and Victoria Z Goldenshtein and Elizabeth W Kahuno and Purav P Vagadia and Marie H Loughran and Lei Zhiquan and Mark E McDonnell and Miranda L Scalabrino and Mishek Thapa and Tammy M Hawley and Greg D Field and Court Hull and Gary E Schiltz and Lindsey L Glickfeld and Allen B Reitz and Michael R Tadross},

url = {https://pubmed.ncbi.nlm.nih.gov/38877316/},

doi = {10.1038/s41592-024-02292-9},

issn = {1548-7105},

year  = {2024},

date = {2024-07-01},

urldate = {2024-07-01},

journal = {Nat Methods},

volume = {21},

number = {7},

pages = {1288--1297},

abstract = {Precision pharmacology aims to manipulate specific cellular interactions within complex tissues. In this pursuit, we introduce DART.2 (drug acutely restricted by tethering), a second-generation cell-specific pharmacology technology. The core advance is optimized cellular specificity-up to 3,000-fold in 15 min-enabling the targeted delivery of even epileptogenic drugs without off-target effects. Additionally, we introduce brain-wide dosing methods as an alternative to local cannulation and tracer reagents for brain-wide dose quantification. We describe four pharmaceuticals-two that antagonize excitatory and inhibitory postsynaptic receptors, and two that allosterically potentiate these receptors. Their versatility is showcased across multiple mouse-brain regions, including cerebellum, striatum, visual cortex and retina. Finally, in the ventral tegmental area, we find that blocking inhibitory inputs to dopamine neurons accelerates locomotion, contrasting with previous optogenetic and pharmacological findings. Beyond enabling the bidirectional perturbation of chemical synapses, these reagents offer intersectional precision-between genetically defined postsynaptic cells and neurotransmitter-defined presynaptic partners.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}