DREADD Agonist 21 Is an Effective Agonist for Muscarinic-Based DREADDs <i>in Vitro</i> and <i>in Vivo</i>

Thompson, Karen; Khajehali, Elham; Bradley, Sophie J.; Navarrete, Jovana; Huang, Xi Ping; Slocum, Samuel T.; Jin, Jian; Liu, Jing; Xiong, Yan; Olsen, Reid H.J.; DiBerto, Jeffrey F.; Boyt, Kristen M.; Pina, Melanie M.; Pati, Dipanwita; Molloy, Colin; Bundgaard, Christoffer; Sexton, Patrick M.; Kash, Thomas L.; Krashes, Michael J.; Christopoulos, Arthur; Roth, Bryan L.; Tobin, Andrew B.

doi:10.1021/acsptsci.8b00012

Cited by 146 publications

(134 citation statements)

References 51 publications

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“…Since virus transduction and expression profile is different between non-human primates and rodents, the expression profile of AAV-PHP.S needs to be validated in non-human primates (36). Additional work is also required to engineer more PNS specific AAVs and to optimize DREADDs (37) and their corresponding ligands (38) for increasing transduction efficiency and regulating dosing.…”

Section: Discussionmentioning

confidence: 99%

Intra-articular AAV-PHP.S mediated chemogenetic targeting of knee-innervating dorsal root ganglion neurons alleviates inflammatory pain in mice

Chakrabarti

Pattison

Doleschall³

et al. 2020

Preprint

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24Objective: 25 Joint pain is the major clinical symptom of arthritis that affects millions of people. 26Controlling the excitability of knee-innervating dorsal root ganglion (DRG) neurons (knee 27 neurons) could potentially provide pain relief. Therefore, our objective was to evaluate 28 whether the newly engineered adeno-associated virus (AAV) serotype, AAV-PHP.S, can 29 deliver functional artificial receptors to control knee neuron excitability following intra-30 articular knee injection. 31 Methods: 32AAV-PHP.S virus packaged with dTomato fluorescent protein and either excitatory (Gq) or 33 inhibitory (Gi) designer receptors activated by designer drugs (DREADDs) was injected 34 into the knee joint of adult mice. Labelling of DRG neurons by AAV-PHP.S from the knee 35 was evaluated using immunohistochemistry. Functionality of Gq-and Gi-DREADDs was 36 evaluated using whole-cell patch clamp electrophysiology on acutely cultured DRG 37 neurons. Pain behavior in mice was assessed using a digging assay, dynamic weight bearing 38 and rotarod, before and after intra-peritoneal administration of the DREADD activator, 39Compound 21. 40 Results: 41We show that AAV-PHP.S can deliver functional genes into the DRG neurons when 42 injected into the knee joint in a similar manner to the well-established retrograde tracer, fast 43 blue. Short-term activation of AAV-PHP.S delivered Gq-DREADD increases excitability 44 of knee neurons in vitro, without inducing overt pain in mice when activated in vivo. By 45 contrast, in vivo Gi-DREADD activation alleviated complete Freund's adjuvant mediated 46 3 knee inflammation-induced deficits in digging behavior, with a concomitant decrease in 47 knee neuron excitability observed in vitro. 48 Conclusions 49We describe an AAV-mediated chemogenetic approach to specifically control joint pain, 50 which may be utilized in translational arthritic pain research.

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Section: Discussionmentioning

confidence: 99%

Intra-articular AAV-PHP.S mediated chemogenetic targeting of knee-innervating dorsal root ganglion neurons alleviates inflammatory pain in mice

Chakrabarti

Pattison

Doleschall³

et al. 2020

Preprint

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“…The chemical structure, potency values (EC 50 /pEC 50 ) and binding affinities (K i /pK i ) of currently used DREADDs agonists: clozapine‐ N ‐oxide, clozapine, compound 21, JHU37152, JHU37160, JHU37107, perlapine, olanzapine, varenicline, and salvinorin B. References: (1) Bonaventura et al (), (2) Vardy et al (), (3) Thompson et al (), (4) Jendryka et al (), (5) Magnus et al (), (6) Weston et al ()…”

Section: Introductionmentioning

confidence: 99%

“…Given the aforementioned unfavourable characteristics of CNO, recent efforts led to screening of several novel compounds that have been suggested as potent DREADD‐ligands that could replace CNO (Chen et al, ). One of the first CNO alternatives, compound 21 (C21, Figure ), displayed greater selectivity for activating hM3Dq than the native muscarinic receptors (Chen et al, ) and has been claimed to be an effective agonist for muscarinic‐based DREADDs both in vitro and in vivo (Thompson et al, ). However, Bonaventura et al recently demonstrated that C21 has low affinity and potency towards DREADDs in vivo and poor BBB penetrance (Bonaventura et al, ).…”

Section: Introductionmentioning

confidence: 99%

Combining designer receptors exclusively activated by designer drugs and neuroimaging in experimental models: A powerful approach towards neurotheranostic applications

Peeters

Missault

Keliris

et al. 2020

British J Pharmacology

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The combination of chemogenetics targeting specific brain cell populations with in vivo imaging techniques provides scientists with a powerful new tool to study functional neural networks at the whole‐brain scale. A number of recent studies indicate the potential of this approach to increase our understanding of brain function in health and disease. In this review, we discuss the employment of a specific chemogenetic tool, designer receptors exclusively activated by designer drugs, in conjunction with non‐invasive neuroimaging techniques such as PET and MRI. We highlight the utility of using this multiscale approach in longitudinal studies and its ability to identify novel brain circuits relevant to behaviour that can be monitored in parallel. In addition, some identified shortcomings in this technique and more recent efforts to overcome them are also presented. Finally, we discuss the translational potential of designer receptors exclusively activated by designer drugs in neuroimaging and the promise it holds for future neurotheranostic applications.

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“…However, drawbacks of CNO include poor brain penetrance, action as 74 a substrate for P-glycoprotein, and reverse metabolism to its parent compound (Gomez et deschloroclozapine -possess better brain penetrance and higher affinity and potency for 81 DREADD receptors than CNO. All of these actuators have been tested in rodent systems 82 (Gomez et al, 2017; Thompson et al, 2018;Weston et al, 2018) and low-dose clozapine 83 (Raper et al, 2019) and deschloroclozapine (Nagai et al, 2019) have been tested in nonhuman 84 primates. Clozapine and olanzapine are atypical antipsychotics designed for clinical use, and 85 deschloroclozapine is a metabolite of clozapine; each has high affinity for DREADD receptors 86 (Gomez et al, 2017;Weston et al, 2018;Nagai et al, 2019).…”

Section: Introduction 66mentioning

confidence: 99%

“…These drugs 308 exhibit greater DREADD activation than clozapine at lower concentrations, indicating that they 309 are more potent agonists at DREADD receptors. In contrast, another actuator drug, Compound 310 21, exhibits similar potency to CNO at the DREADD receptor; however, this potency is lower 311 than both J52 and J60 and thus may not be practical for applications in nonhuman primates 312 (Chen et al, 2015;Thompson et al, 2018;Bonaventura et al, 2019). Recently, it was reported 313 that low-dose Compound 21 also produced metabolic changes in brain activity (FDG uptake) 314 while an equipotent dose of clozapine produced no changes (Bonaventura et al, 2019).…”

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confidence: 99%

Chemogenetic actuator drugs impair prefrontal cortex-dependent working memory in rhesus monkeys

Upright

Baxter

2019

Preprint

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27The most common chemogenetic neuromodulatory system, Designer Receptors Exclusively 28 Activated by Designer Drugs (DREADDs), uses a non-endogenous actuator ligand to activate a 29 modified muscarinic acetylcholine receptor that is no longer sensitive to acetylcholine. It is 30 crucial in studies using these systems to test the potential effects of DREADD actuators prior to 31 any DREADD transduction, so that effects of DREADDs can be attributed to the chemogenetic 32 system rather than the actuator drug. We investigated working memory performance after 33 injections of three DREADD agonists, clozapine, olanzapine, and deschloroclozapine, in male 34 rhesus monkeys tested in a spatial delayed response task. Performance at 0.1 mg/kg clozapine 35 and 0.1 mg/kg deschloroclozapine did not differ from mean performance after vehicle in any of 36 the four subjects. Administration of 0.2 mg/kg clozapine impaired working memory function in 37 three of the four monkeys. Two monkeys were impaired after administration of 0.1 mg/kg 38 olanzapine and two monkeys were impaired after the 0.3 mg/kg dose of deschloroclozapine. We 39 speculate that the unique neuropharmacology of prefrontal cortex function makes the primate 40 prefrontal cortex especially vulnerable to off-target effects of DREADD actuator drugs with 41 affinity for endogenous monoaminergic receptor systems. These findings underscore the 42 importance of within-subject controls for DREADD actuator drugs to confirm that effects 43 following DREADD receptor transduction are not due to the actuator drug itself, as well as 44 validating the behavioral pharmacology of DREADD actuator drugs in the specific tasks under 45 study. 46 47 48 49 50 51 52 Significance Statement 53 Chemogenetic technologies, such as Designer Receptors Exclusively Activated by Designer 54Drugs (DREADDs), allow for precise and remote manipulation of neuronal circuits. In the 55 present study, we tested monkeys in a spatial delayed response task after injections of three 56 actuator drugs -clozapine, olanzapine, and deschloroclozapine. We found that monkeys 57 showed significant working memory impairments after 0.2 mg/kg clozapine, 0.1 mg/kg 58 olanzapine, and 0.3 mg/kg deschloroclozapine compared to vehicle performance. In monkeys 59 that showed impairments, these deficits were particularly apparent at longer delay periods. It is 60 imperative to validate the drugs and dosages in the particular behavioral test to ensure any 61 behavior after DREADD transduction can be attributed to activation of the receptors and not 62 administration of the actuator drug itself. 63 64

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DREADD Agonist 21 Is an Effective Agonist for Muscarinic-Based DREADDs in Vitro and in Vivo

Cited by 146 publications

References 51 publications

Intra-articular AAV-PHP.S mediated chemogenetic targeting of knee-innervating dorsal root ganglion neurons alleviates inflammatory pain in mice

Intra-articular AAV-PHP.S mediated chemogenetic targeting of knee-innervating dorsal root ganglion neurons alleviates inflammatory pain in mice

Combining designer receptors exclusively activated by designer drugs and neuroimaging in experimental models: A powerful approach towards neurotheranostic applications

Chemogenetic actuator drugs impair prefrontal cortex-dependent working memory in rhesus monkeys

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