Noninvasive focal gene transfer of chemogenetic proteins in the primate brain.

The development of chemogenetic neuromodulators, including Designer Receptors Exclusively Activated by Designer Drugs (DREADDs), have enabled focally specific, long-lasting, and reversible neuromodulation in the primate brain. Although systemically delivered synthetic ligands allow for noninvasive actuation of chemogenetic receptors, direct intraparenchymal injection remains atop the available methods to precisely deliver chemogenetic payloads to a specific target of the brain. The requirement of trephination, however, is accompanied by inherent risks of infection, long recovery times, and often tissue damage with concomitant behavioral complications. When considering therapeutic injections, the requirement of transcranial surgery does not translate well to the clinic, especially when repeated administrations are required. Here, we leverage our recent development of transcranial focused ultrasound (tFUS) for noninvasive and focal delivery of adeno-associated viruses (AAVs) carrying excitatory Gq-DREADDs to frontal cortical targets (areas 6DR and 8aD) in the marmoset brain. Using [ ¹⁸ F]-fluorodeoxyglucose (FDG) positron emission tomography, we demonstrate significant increases in glucose metabolism at the site of viral delivery after administering the DREADD-specific agonist deschloroclozapine (DCZ), as compared to vehicle control. Focal neuronal DREADD expression was confirmed by immunohistochemistry at the site of opening. Through comparison of awake resting-state functional connectivity (whole brain connectivity with the sites of delivery) and structural connectivity (directly injected viral neuronal tracing at the sites of delivery) we demonstrate that the increase in glucose metabolism occurs at both mono- and polysynaptically connected brain regions. Taken together, these results demonstrate the ability to focally deliver excitatory chemogenetics without the need for surgery, allowing for activation of long-range frontal cortex circuits of the primate brain.