Investigations into extended amygdalar cholecystokinin for cocaine-related sleep disturbances and sleep-dependent cocaine memories

The National Institute on Drug Abuse awarded The Leland Stanford Junior University $153,560 on January 19, 2027, under the Drug Use and Addiction Research Programs (CFDA 93.279) to investigate the role of cholecystokinin-expressing neurons in the bed nucleus of the stria terminalis in regulating sleep homeostasis and sleep-dependent cocaine memories. The research examines how cocaine withdrawal induces sleep disturbances and how dysregulated sleep patterns contribute to impaired decision-making and persistent substance abuse. The study employs bulk calcium imaging to measure in vivo activity of these neurons during cocaine withdrawal while simultaneously recording sleep-wake patterns via polysomnography, with planned chemogenetic inhibition experiments in subsequent phases. Performance occurs in Palo Alto, California, with a period of performance extending through August 31, 2029. This is a Project Grant, a form of assistance that supports basic and translational research in substance use prevention, treatment, and addiction outcomes.

Abstract

Sleep is an integral homeostatic process that allows for physiological recovery and supports emotional and cognitive processes. Sleep disturbances have been shown to produce adverse effects on memory consolidation, which is a critical mechanism that mediates decision-making and other behaviors when awake. Dysregulated sleep patterns are commonly seen in individuals with a substance use disorder. IT is also well-established that substance dependence is, in part, the result of maladaptive decisions to seek and take drugs. Thus, impairments in decision-making seen in substance use disorders that lead to aberrant drug use may potentially be due to disturbances in sleep homeostasis and sleep-mediated drug memories. Identifying a multifunctional neural substrate involved in regulating sleep, memory, and substance use would be key to furthering our understanding of how these behavioral interactions give rise to persistent substance abuse and inform future treatment approaches. The bed nucleus of stria terminalis (bnst) contains a variety of distinct, neuropeptide-expressing neurons that serve functions in a range of behaviors, including stress and emotional arousal. The proposed study will investigate the involvement of a heavily understudied subpopulation of gabaergic neurons in the bnst, those expressing cholecystokinin (cckbnst), for regulating sleep homeostasis and sleep-dependent drug memories. Using bulk calcium imaging to measure in vivo activity of cckbnst neurons aligned to simultaneous polysomnographic recordings to monitor sleep-wake patterns, the first aim will examine the activity profile of cckbnst neurons during cocaine withdrawal-induced dysregulated sleep, and how these sleep and neuronal activity dynamics evolve across discrete withdrawal time points. The second aim will employ chemogenetic inhibition of cckbnst neurons to determine their necessity in promoting sleep under normal and disrupted sleep homeostasis. This will reveal a potential mechanism by which cckbnst neurons are contributing to sleep-wake regulation. The third and final aim will elucidate the involvement of cckbnst neurons in cocaine-seeking via sleep-dependent drug memories by examining neuronal reactivation patterns following exposure to a cocaine- related context and using targeted optogenetic manipulations during sleep-mediated memory consolidation to either strengthen or weaken cocaine seeking. Collectively, the outcomes from this proposal will provide novel and valuable insight about the role of cckbnst neurons in maladaptive drug seeking and serve as impetus for further research characterizing this promising neural substrate in the development of treatments for substance dependence, targeting sleep disturbances and memory processes underpinning CUE-induced relapse.