NIAAA - National Institute on Alcohol Abuse and Alcoholism Grants

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Excessive alcohol consumption is thought to be the third leading preventable cause of death in the United States. But these estimates of the deleterious effect of alcohol are largely based off self-reports of alcohol consumption. Concerningly, because we and others have shown that clinical self-reports are unreliable, while others have shown that the estimates of daily alcohol consumption by CDC are off by at least three-fold, the true cost of excessive drinking may be much higher. In fact, it is possible that excessive alcohol consumption is the leading preventable cause of death. We have developed a methylation sensitive digital PCR (MSdPCR) test called the Alcohol T Score (ATS) that quantifies chronic alcohol consumption. In several peer reviewed studies, we have shown that the ATS significantly outperforms the carbohydrate deficient transferrin (CDT) test, the current industry gold standard, in predicting chronic heavy alcohol consumption (HAC; ≥ 6 drinks/day for 8 weeks) and strongly predicts death in those with severe coronary heart disease. We recently examined an epidemiologically sound sample of DNA and data from 4910 subjects (3:1 mix of lung cancer cases and controls) who participated in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial with the ATS. We found that the ATS was a powerful predictor of mortality in smokers with (n=92) and without lung cancer (n=402). In fact, in both groups, the effects of alcohol consumption on mortality greatly outweighed those of smoking. Taken together with our prior findings, these new findings strongly suggest that heavy drinking, not smoking is the largest preventable cause of mortality in the United States. Unfortunately, this sample was largely White and nonsmoking, which limits generalizability of the results. Therefore, in this high-risk R43 study, we propose to extend our ATS determinations to all non-smoking subjects in this group (n=612) and all of those of non-White ancestry (n=985) from our existing cohort of 5000 subjects from the PLCO cohort. We hypothesize that the ATS will predict mortality broadly across both sexes, and in all ancestries whether or not they smoke. This project is innovative because heavy alcohol consumption (HAC) is often missed by clinicians. It is high impact because HAC may be a significant, if not the largest preventable cause of death in the United States. The team is well prepared to conduct the research including statisticians and a leading expert on DNA methylation. It is highly feasible because the assay already exists and we have both the DNA and the data. As a direct result of this research, we will determine the frequency and effect of HAC on a broad epidemiologically sound, cross section of the American public. As an indirect outcome, we will demonstrate the potential use case for conducting this rapid, easy to perform MSdPCR test as part of medical assessments of those presenting for lung or perhaps any other cancer screening while gathering needed data for eventual approval of the ATS by the FDA as a test for HAC.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Scaling up AUD Information Resources for Provider Education: The Behavioral Health Counseling Competencies AUD Training Suite Project Summary The alcohol treatment gap is a multi-dimensional phenomenon that requires intervention related to public awareness, access to specialty care, and improvement in the quality and quantity of the broader, behavioral health workforce. The current proposal targets the latter via the development of a training product designed to increase provider knowledge and skill related to alcohol misuse and AUD. This Phase I/II Fast Track STTR proposal will create a didactic and interactive simulation training that will facilitate provider use of the content and tools within NIAAA’s key dissemination resource - Rethinking Drinking. The product will incorporate two additional elements. First, content delivery will be grounded in an NIAAA-funded training program targeting four core competencies of behavioral health counseling: 1) developing a behavior change relationship, 2) providing psychoeducation, 3) setting and monitoring behavior change goals, and 4) providing behavioral skills training. Second, training technology will include proprietary conversation-simulation technology that has a 15-year track record in changing provider behaviors. The team includes the small business applicant, the primary academic partner, and the conversation simulation company. Additional content expertise will be provided by a panel of subject matter experts. The team are established independent investigators, small business award recipients, and long-standing collaborators, which enhances the feasibility and probability of success in meeting the following Specific Aims. Phase I will establish feasibility and technical merit via the development, expert panel review, and end user piloting of the first of the four-module training program – Alcohol and you, establishing a behavior change relationship. Phase II will develop Module 2 How much is too much? providing psychoeducation, Module 3 Thinking about change, goal setting and monitoring, and Module 4 Setting drinking boundaries, providing behavioral skills training and conduct a pilot randomized trial (N = 55) comparing the training suite to a dissemination as usual control using a pre-post, standardized patient design. Commercialization will occur throughout the award period and particularly in year three. The proposed project merges team expertise and technical support to create a novel training product that can increase the quality and quantity of the behavioral health workforce, thereby contributing to reductions in the alcohol treatment gap.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT SUMMARY / ABSTRACT Hazardous alcohol consumption remains a significant public health issue in the U.S., responsible for over 140,000 deaths annually. Despite its profound impact, only about 10% of individuals diagnosed with alcohol use disorder (AUD) receive any form of treatment, underscoring a substantial treatment gap. This SBIR Phase I proposal seeks to address this gap by enhancing the "Step Away Alcohol Manager" (SAAM), a mobile health (mHealth) intervention developed by Here and Now Systems, LLC (HNS). SAAM builds on the success of the Step Away app, which has demonstrated significant reductions in alcohol use —50-60% reductions in four pilot trials. However, sustaining long-term user engagement remains a challenge, with only 40% of users continuing to engage with the app after six months and some modules being used infrequently. This project aims to integrate cutting-edge artificial intelligence (AI) and biometric technologies to enhance personalization, improve engagement, and increase the app’s overall effectiveness in promoting long-term behavior change. Specifically, we will incorporate a multi-agent generative AI (GenAI) and large language model (LLM) to provide users with empathic, real-time conversations and strategies to manage drinking-related challenges such as difficulty sleeping, stress, and cravings. The AI will initiate interactions during users' high-risk times for drinking and during daily check-ins, providing proactive and responsive support. Additionally, we will integrate facial scan technology which will monitor users’ stress levels, sleep quality, and overall health in real time. This biometric data will be used to tailor feedback and enhance the relevance of the AI-driven interactions, further personalizing the user experience. The Phase I project has three specific aims: (1) Develop SAAM with advanced AI-driven conversational capabilities and real-time biometric feedback. The chatbot will deliver evidence-based responses based on trusted sources, ensuring it adheres to scientifically sup ported interventions. (2) Conduct end-user testing with 20 participants who meet AUDIT criteria for hazardous drinking to evaluate the app’s content, usability, and the quality of the personalized messages generated by the A I. We will use validated tools such as the Client Satisfaction Questionnaire (CSQ) and System Usability Scale (SUS), with benchmarks of 24+ on the CSQ and 70+ on the SUS. We will also utilize qualitative research methodology to determine themes related to SAAM’s strengths and limitations, which will be used to inform needed modifications. (3) Refine SAAM based on feedback from these reviews, addressing any usability issues, content gaps, and engagement barriers to create a polished, user-friendly app ready for Phase II evaluation. Through this Phase I project, SAAM has the potential to significantly expand access to effective, evidence-based interventions for alcohol use disorder by leveraging advanced AI and biometric technologies to provide real-time, personalized support. This innovative approach will address the critical public health need for more accessible and engaging alcohol treatment solutions, offering a novel, scalable intervention with the potential for widespread adoption.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Abstract: Alcohol consumption adversely affects up to 28% of hospitalized patients and contributes to a loss of 133 million disability-adjusted life years and 5.3% of worldwide deaths each year. As alcohol use has substantial health and economic impact, much attention has been directed toward the numerous adverse health outcomes in patients with unhealthy alcohol use. Alcohol use disorder (AUD) occurs on a spectrum from mild to severe and is precipitated by binge drinking and heavy alcohol use. We and others have shown that AUD is a modifiable perioperative risk factor and is present in up to 18% of surgical patients. Despite its significant clinical impact, AUD is often overlooked in the design of perioperative care plans. Although AUD affects 9% of the US population, less than one in ten individuals with AUD receives any treatment. Barriers to treatment are multiple and include scarce care availability, limited access, and social stigma. Although anesthesiologists routinely provide guideline-concordant treatment for non-operative medical conditions such as coronary artery disease, AUD-specific care is rarely provided, even to high-risk patients. Thus, there is a compelling opportunity to integrate AUD screening and treatment into routine perioperative care. Our central hypotheses are that AUD health services initiated in the perioperative period will 1) leverage the significant resources made available to perioperative care in the US, 2) forge novel synergistic alliances between previously disconnected healthcare settings, and 3) break down barriers to access to care for broad populations. A multi-institutional team of anesthesiologists, biostatisticians, psychologists, and psychiatrists with expertise in AUD treatment and novel clinical trial designs will lead the Leveraging Alcohol Use Disorder Screening for Treatment in Routine Perioperative Care: AllUsCare proposal and proposes three aims: 1) Leverage our existing EHR-integrated Alcohol Use Disorders Identification Test-Concise (AUDIT-C) screening tool to increase provision of AUD-specific perioperative care, 2) Conduct a two-center prospective observational cohort study to assess patient acceptability of interventions, feasibility of outcome data collection, and optimum outcome measures for a future pragmatic trial, and 3) Optimize perioperative AUD intervention bundles most likely to be effective in a future pragmatic randomized factorial cluster trial. This R34 planning grant will lay the groundwork for identifying the most effective health service intervention bundles in surgical patients at high risk for AUD. To maximize inclusion of broad populations, we will conduct our study in two centers that serve inner-city and rural populations and will deliberately include patients aged 12 years and older. At the conclusion of AllUsCare, we will have established the research team, designed an AUD intervention bundle most likely to be effective in a future multi-center pragmatic trial, demonstrated a single-IRB governed uniform data collection and entry process, and confirmed the acceptability and feasibility of the future pragmatic trial.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Project Summary/Abstract American Indian and Alaska Native (AIAN) communities experience significant alcohol-related problems, further propagated by obstacles to beginning effective, high-quality treatment for alcohol use disorder (AUD). Precision medicine approaches to AUD treatment may make treatment more efficient and effective by individually tailoring AUD treatments to common etiological and maintenance mechanisms underlying AUD. For example, recent precision medicine efforts have identified that individuals who drink primarily in the context of reward (i.e., drinking for enhancement and social interaction) may respond better to naltrexone, whereas those who drink primarily in the context of relief (i.e., drinking to relieve negative physical and affective states) may respond better to acamprosate. However, extant measures used to classify reward and relief drinking tendencies do not account for the contextual factors associated with these constructs in AIAN peoples. Identifying relevant measures of reward and relief drinking tendencies in AIAN peoples may inform precision medicine research and reduce burden related to alcohol problems in AIAN. The proposed work will utilize a multi-method approach grounded in a community-engaged research framework to develop a tailored measure of reward and relief drinking by refining and expanding on several existing measures of reward and relief drinking and assess the construct validity of the newly developed measure in AIAN peoples. First, we will recruit AIAN people with AUD (N=20) to examine the content validity of three existing measures of reward and relief drinking and use cognitive interviewing methods to refine existing items and/or develop additional relevant items. Second, we will administer these items and any participant-identified additional items to a geographically varied sample (N=100) of AIAN with AUD. Using these data, we will then examine structural validity by identifying the hierarchical contributions of individual items to respective reward and relief drinking factors by conducting nonparametric item response theory (IRT) analyses. Lastly, we will utilize the refined measure to identify reward and relief drinking subgroups and examine comparative associations with convergent constructs to assess external validity. Ultimately, the proposed research will produce a construct-valid measure of reward and relief drinking informed by the lived experiences of AIAN peoples with AUD. The tailoring of an AIAN-specific measure of reward and relief drinking will play a direct role in improving alcohol problems in this population.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Project Summary Alcohol use disorder (AUD) is a prevalent, affecting over 10% of Americans, with only a fraction of individuals seeking treatment. Chronic alcohol exposure induces maladaptive changes in neuronal physiology, particularly within neural circuits that mediate reward and consumption behaviors, hallmarks of AUD that render treatment difficult and increases the likelihood of relapse. Despite substantial progress in understanding neurotransmitter- mediated changes in AUD, the role of neuropeptides such as orexin (hypocretin) remains underexplored. Orexin is a hypothalamic neuropeptide that contributes to a number of physiological conditions, including regulation of arousal, feeding, and stress. Orexinergic neurons from the lateral hypothalamic area (LHA) innervate a number of brain regions implicated in AUD, such as the ventral tegmental area (VTA) and nucleus accumbens (NAc). In my preliminary data, I have found that chronic voluntary EtOH consumption leads to activation of orexinergic neurons in the LHA in male and female C57BL/6J mice. Furthermore, I have demonstrated that I can track orexinergic activity in a free-moving mouse while it is consuming EtOH through the use of fiber photometry and the orexin peptide sensor, OxLight1. This proposal aims to investigate the impact of alcohol dependence on orexinergic circuits, particularly the LHA-VTA pathway, and its role in excessive alcohol consumption. Using cutting-edge tools, including the OxLight1 biosensor for real-time orexin monitoring, optogenetics for pathway- specific neuronal silencing, and whole-cell patch-clamp electrophysiology, this study will elucidate the functional changes in orexinergic activity in male and female mice following chronic intermittent ethanol (CIE) exposure. The innovative approach includes sex-specific and within subject analyses and behaviorally guided experiments to address critical gaps in the field. Aim 1 of this proposal will determine how silencing LHA orexinergic neurons projecting to the VTA modulates voluntary alcohol consumption in ethanol-dependent mice. Aim 2 of this proposal will assess functional adaptations in the LHA orexinergic circuits to regions implicated in AUD following ethanol dependence and their impact on voluntary alcohol consumption. The expected outcomes will reveal how chronic alcohol exposure alters orexinergic signaling, contributing to excessive drinking and relapse. These findings will enhance our understanding of AUD's neural mechanisms, identify potential sex differences, and detect therapeutic strategies targeting orexinergic pathways. This research holds significant translational potential, advancing efforts to mitigate the global burden of AUD.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Project Summary/Abstract This project proposes to investigate the underlying neural mechanisms within the nucleus accumbens (NAc) driving cue-elicited reward seeking during aversion-resistant alcohol drinking (ARD), defined as drinking despite negative consequences or “compulsive alcohol drinking”, using optogenetics and simultaneous in-vivo electrophysiology and optogenetics in male and female Long-Evans rats. Environmental reward predicting cues provide a source of motivation for reward-seeking which may be heightened following extended alcohol use, resulting in maladaptive, ARD, a defining feature of alcohol use disorder (AUD). Despite the well- established role of the NAc in mediating cue-elicited reward seeking, motivation, and ARD, the neural signature within the NAc during cue and alcohol access in ARD is not known, which greatly limits prevention and treatment of AUD. The NAc is innervated by glutamatergic projections from the paraventricular nucleus of the thalamus (PVT). This projection is thought to be relevant for processing motivational conflict and preventing unproductive reward seeking, while sparing normal affective behavior. However, the PVT-to-NAc pathway’s role in modulating NAc encoding during ARD or how the activity of PVT-to-NAc neurons alter ARD is not known. Thus, the central hypotheses of this project are 1) the NAc encodes cue and outcome information related to ARD. Specifically, neural activity in the NAc responds to reward (alcohol access) predictive cues and to alcohol rewards in a discriminative stimulus task and that over continued alcohol use, this activity increases in correlation with the development of ARD and 2) the neuronal activity in the NAc is causally related to an excitatory projection from the PVT that acts to prevent ARD initially, but erodes over time resulting in ARD. This project will assess the neural mechanisms of cue-elicited reward seeking and ARD during a discriminative stimulus task using both optogenetic circuit manipulations and simultaneous optogenetics and awake-and- behaving electrophysiology recordings. The training plan for this project is curated in an ideal research environment in the Department of Neuroscience at the University of Minnesota which will provide training in cutting edge neuroscience techniques, professional development, and research ethics that will amass an ideal training experience to facilitate my career as an independent alcohol research scientist. The specific research hypotheses are 1) Optogenetic inhibition of PVT-to-NAc neurons at cue presentation and alcohol reward will cause ARD in otherwise aversion-sensitive rats. 2) Optogenetic excitation of PVT-to-NAc at cue presentation and alcohol reward will reduce ARD, causing aversion-sensitivity, in otherwise ARD rats. 3) Ensembles of neurons within the NAc encode cue and alcohol rewards during ARD. This encoding is facilitated by the PVT- to-NAc projection whereby PVT-to-NAc inhibition will increase NAc encoding of cues and alcohol rewards and PVT-to-NAc excitation will reduce this encoding within the NAc. Completion of the proposed work will elucidate understanding of the neural mechanisms of ARD and will aid in the prevention and treatment of AUD.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT SUMMARY The ability to adaptively and appropriately respond to changes in the environment is a crucial behavior. It is cognitive flexibility that enables this adaptive responding. An inability to adaptively shift behavior (cognitive inflexibility) despite negative outcomes is observed in various psychiatric disorders and is a key diagnostic criterion of alcohol use disorder (AUD). The long-term objective of this work is to elucidate how chronic alcohol and stress promote AUD through cognitive inflexibility. In humans, AUD is accompanied by aberrant frontal cortex function alongside deficits in cognitive flexibility. A significant body of work, including that from our lab, has demonstrated impaired cognitive flexibility in animal models following chronic alcohol which can be exacerbated by stress exposure. However, the frontal cortical neuroadaptations that mediate these alcohol and stress induced cognitive impairments remain unresolved. This is a critical gap given that stress is a key risk factor in perseverative relapse for individuals with AUD. My central hypothesis is that functional network changes across the dorsomedial frontal cortex underlie cognitive flexibility impairments following repeated alcohol and stress exposure. Moreover, I hypothesize that changes in modulatory norepinephrine inputs to this region are critical contributors to these flexibility deficits. To test these hypotheses, I will investigate the mechanisms underlying the negative impact of chronic alcohol and stress on cognitive flexibility using a novel attentional set shifting task. In Aim 1 I will use in vivo calcium imaging to investigate alcohol- and stress- associated changes in dorsomedial frontal cortex dynamics underlying behavioral impairment. In Aim 2, I will determine whether chronic alcohol and stress functionally disrupts noradrenergic regulation of the dorsomedial frontal cortex using in situ hybridization readouts and pharmacological interventions to recover cognition. These proposed studies will provide ample training in cutting-edge techniques and facilitate my professional development in the alcohol research field. At the same time, this work will advance our understanding of the impact of long-term alcohol and stress exposure on cortical function and identify potential mechanisms for therapeutic interventions.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT ABSTRACT Fetal alcohol spectrum disorders (FASD) can occur in a person when they are exposed to alcohol before birth and are characterized by impairments in physical, cognitive, and behavioral functions. Common signs and symptoms associated with FASD include low body weight, abnormal facial features, poor coordination, hyperactive behaviors, difficulty with attention, and difficulty in school. There is also growing research that individuals with FASD have sensory processing deficits that can impact self-regulation and adaptive behavior. Sensory enriched occupational therapy intervention may improve the neurological sensory processing deficits experienced by individuals with FASD, which may lead to lifelong improvements in self-regulation and adaptive behavior. Here, we propose a Phase 1 clinical trial to assess the following: 1) characterize changes in brain white matter microstructure of sensory integration in 6 to 8-year old children with FASD following intervention; 2) characterize changes in sensory processing, self-regulation, and adaptive behavior; and 3) develop a conceptual model of mediation, whereby a sensory enriched intervention alters brain sensory white matter networks, leading to improvements in sensory processing, self-regulation, and adaptive behavior in children with FASD. We predict that children with FASD who receive a sensory enriched intervention will demonstrate positive changes in white matter neural architecture when compared to controls, and that we will see improvements in sensory processing, self-regulation, and adaptive behaviors. This two phase R61/R33 will allow us the time to adapt and assess the study procedures and intervention to inform the development of future larger clinical trials.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Project Summary This interdisciplinary project addresses the pressing public health challenge of alcohol use disorder (AUD). AUD affects approximately 14% of adults in the United States alone, with profound societal and individual detrimental consequences. Despite its prevalence, effective treatments for AUD are limited, highlighting the urgent need for a better understanding of the underlying molecular mechanisms of this disorder. This project investigates the role of neurexin 3 (NRXN3) in AUD using rhesus macaque and mouse models. Our laboratory has shown that NRXN3, a presynaptic transmembrane protein, is downregulated in the prefrontal cortex of high ethanol consuming non-human primates. Given the central role of this protein in synaptic transmission and plasticity, processes known to underlie AUD-related behaviors, investigating its role in mediating the effects of alcohol in brain function is paramount. However, the specific role of NRXN3 in AUD remains poorly understood, with only a handful of published GWAS studies linking single nucleotide polymorphisms with different risk propensities for addiction. Here, we aim to elucidate the NRXNs' landscape in the cortex following chronic ethanol use and protracted abstinence, with particular focus on a specific transcript/isoform, Nrxn3β, and its role in synaptic plasticity. It also explores manipulating Nrxn3β transcripts in one of the major inhibitory neurons in the cortex, parvalbumin (PV) neurons, to modulate excitatory outputs to reward and motor centers and reduce ethanol intake. By employing cutting-edge molecular profiling techniques, advanced immunohistological analyses, and in vivo manipulation, we will examine changes in Nrxns’ expression and excitatory/inhibitory synaptic properties in key brain regions implicated in AUD, such as the prefrontal cortex. This research holds significant intellectual significance as it seeks to advance our understanding of the neurobiological basis of AUD and identify novel therapeutic targets. By uncovering the precise mechanisms through which Nrxn3β influences synaptic plasticity and alcohol-related behaviors, we aim to inform the development of targeted interventions to mitigate the adverse effects of AUD and improve treatment outcomes for affected individuals.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT SUMMARY/ABSTRACT Early life stress is a prominent risk factor for initiation of alcohol use and increased lifetime alcohol consumption. Both adolescent stress and alcohol use increase the risk for developing depression during adulthood, and both exposures negatively influence the neuroimmune system, resulting in a sensitized or exaggerated response to a neuroimmune challenge. However, in rodent models, the interactive effects of alcohol and stress co-exposure are not widely studied. Because there is a bidirectional relationship among adolescent stress and alcohol use, it is critical for these exposures to be studied independently as well as simultaneously to capture synergistic or opposing effects. Therefore, one overall goal of the current proposal is to utilize an exposure paradigm that captures both the independent effects of adolescent intermittent ethanol (AIE) and chronic corticosterone (CORT) consumption in the drinking water, a commonly used model to mimic the downstream effects of one primary component of chronic stress by a tractable means. One component of the neuroimmune system that sits at the nexus between chronic stress and ethanol exposure is the inflammasome, nucleotide-binding oligomerization domain-like (NOD-like) receptor protein 3 (NLRP3). The NLRP3 inflammasome is highly expressed in the resident immune cell of the brain, microglia, and NLRP3 inflammasome expression is increased following both stress and ethanol exposures, causing a sensitized cytokine response upon challenge. Increased cytokine expression has been implicated in dopaminergic dysfunction, such as decreased dopamine availability and downregulated dopamine receptor expression, which may explain AIE- or stress-induced dysfunction of reward and motivation pathways as well. Therefore, the overall goal of the proposed studies is to determine the independent and interactive effects of AIE and CORT on 1) neuroimmune function, measured by NRLP3 expression and microglial activation state, and 2) changes to the reward and motivational systems including behavioral measures such as sucrose preference for anhedonia and effort-based decision making (EBDM) for motivation, as well as dopaminergic dysfunction such as expression of ΔFosB, D1, D2, and DAT. Specifically, whether the NLRP3 inflammasome plays a mechanistic role as a neuroimmune target for future treatment. Overall, this proposal will provide essential scientific and professional development, so enabling me to reach my goal of becoming an independent researcher.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Abstract Rejuvenation Technologies Inc. (RTI) seeks to continue advancing the translation of a telomere extension biologic, TERT mRNA, which is delivered to the liver intravenously (i.v.) using hepatocyte-targeted lipid nanoparticles (LNPs) for the treatment of alcoholic hepatitis (AH). AH is an acute form of alcoholic liver disease (ALD) that represents a dire unmet medical need, as mortality within 1 month of presentation is 25–50%. Most AH patients exhibit advanced fibrosis/cirrhosis, contributing to acute-on-chronic liver failure. The only definitive treatments are steroids, which are ineffective at reducing patient mortality. However, there is strong evidence that short telomeres play a casual role in AH. Telomeres, the protective DNA tips of chromosomes shorten with each cell division; chronic liver injury due to excessive alcohol consumption drives compensatory hepatocyte proliferation resulting in rapid telomere shortening, hepatocytic senescence, and the secretion of senescence- associated secretory phenotype factors that activate hepatic stellate cells, causing them to become fibrogenic. Short telomeres limit the ability of hepatocytes to divide and the liver to regenerate, ultimately leading to liver failure. RTI’s team has invented a breakthrough treatment addressing shortening telomeres in AH patients comprising 1) the telomere-extending biologic telomerase (TERT) mRNA and 2) an LNP vehicle that delivers mRNA to the liver with high efficiency, transfecting >99% of hepatocytes at very low doses (0.05 mg/kg), with high transfection even in cirrhotic livers. Upon delivery to the cells, TERT mRNA is translated to functional TERT protein, and both are degraded within days. During this brief treatment, telomeres are extended sufficiently to reverse years of telomere shortening. After the TERT degrades, telomeres resume shortening normally. TERT mRNA does not immortalize cells, and TERT is not an oncogene. RTI’s TERT mRNA LNPs are a highly effective, low risk, revolutionary biologic to improve and extend the lives of AH patients. A prior NIAAA Phase II SBIR and an INTERACT meeting on AH with the FDA has led to this proposed Commercialization Readiness Pilot (CRP) that will culminate in an IND submission. The specific aims are: 1) Pharmacokinetic (PK) study of mRNA and select lipids in mice to measure rate of clearance; 2) Pre-treatment evaluation in mice; 3) GMP manufacturing, genotoxicity studies, and IND-enabling GLP toxicology studies in NHPs; and 4) Preparation of IND data package and FDA submission.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Any instances of “women” have been changed to “females”. Information regarding gender discrepancies has been removed. Research has consistently demonstrated that alcohol use is a particularly salient risk factor for sexual assault among college females. Additionally, recent work on alcohol and cannabis co-use, particularly simultaneous co-use (i.e., use on the same occasion so effects are likely to overlap), has indicated that alcohol and cannabis co-use is associated with increased substance use-related negative consequences, including sexual assault victimization and revictimization among college females. However, limited empirical attention has been directed toward integrated alcohol and cannabis co-use and sexual assault protective behavioral strategies (PBS). This F31 application addresses an important gap in the literature in investigating PBS for substance use (alcohol and cannabis co-use) and sexual assault in a deliberately integrated fashion, in that it focuses explicitly on substance use-specific behavior attenuation that may be most useful for college females in mitigating risk for sexual assault. The long-term goal of the proposed research is to inform clinical assessment and intervention research focused on substance-use facilitated sexual assault prevention among young females. This three-phase study will provide the data necessary to develop and refine a novel alcohol and cannabis co-use PBS measure that is specific to the prevention of sexual assault among college females. Specific Aim 1 involves community-based participatory research (CBPR) to gather in-depth qualitative data on how college females use alcohol and cannabis PBS in the context of sexual assault prevention. Specific Aim 2 comprises the pilot testing of initially developed scale items elucidated from Aim 1 to finalize a new measure of co-use-related sexual assault PBS. Lastly, Specific Aim 3 will assess scale performance (i.e., reliability, validity) through investigating the scale’s associations with substance use and sexual assault victimization. This proposed study directly corresponds with NIAAA’s most recent strategic plan for fiscal years 2024-2028 in that its overarching aim is to better understand the effects of alcohol use and misuse among females (i.e., within the context of sexual assault) while informing the development of a tailored PBS measure to address their needs in research and clinical contexts. The proposed award is also consistent with NIAAA’s NOSI (NOT-OD-24-079) on health issues that affect young females, including the etiology, prevention, and treatment of alcohol misuse.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT SUMMARY/ABSTRACT: Adolescence is an important developmental window of heightened, experience-dependent neuroplasticity in brain regions that are involved in essential functions such as cognition. Adolescence is also a time when risky decision-making increases, including alcohol and other substance use, which can have harmful and lasting effects on the developing brain. Further, adverse life experiences in youth can have effects on both neuroplasticity and engagement in alcohol use. The links between adverse life experiences, neurodevelopmental trajectories of plasticity, and problematic alcohol use have not yet been well-defined, thus clouding our mechanistic understanding of these potentially interrelated processes. The proposed project will leverage the Adolescent Brain Cognitive Development Study to clarify these links through three specific aims. In Aim 1, we will explore whether adverse experiences during adolescence lead to alterations in neuroplasticity in cognitive brain regions. We will use latent change score models to examine whether adverse experiences—measured by the Life Events Scale (LES)—lead to later reductions in plasticity, as indicated by the amplitude of low-frequency fluctuations (ALFF), a resting state fMRI-based measure of plasticity. In Aim 2, we will tease apart the direction of impact between neuroplasticity and alcohol use during adolescence. Specifically, we will assess if individual differences in plasticity lead to alcohol use or if alcohol use impacts plasticity. We will apply multilevel and growth curve modeling to alcohol use measures from the Timeline Follow Back (TLFB) survey and to the neuroimaging measure of plasticity (ALFF). In Aim 3, we will explore whether the links between adverse life experiences during adolescence and engagement in alcohol use are causally mediated by neuroplasticity. We will apply causal mediation analyses to adversity data from the LES, alcohol use data from the TLFB, and ALFF plasticity data. Findings from this project will inform a mechanistic understanding of the impacts of early alcohol use on neurodevelopment and on the influence of brain-environment interactions on the etiology of youth alcohol use, thereby providing novel and specific targets for future interventions. This NIAAA F32 project is highly feasible in the proposed timeline and well-supported by the mentorship team (Drs. Lindsay Squeglia and Louise Mewton) and institution (Medical University of South Carolina). The proposed project will provide essential training for the candidate’s long-term career development heading toward an independent research career focused on applying developmental neuroscience methods to the study of adolescent alcohol use disorder (AUD). This study will provide the first step toward generating hypotheses for an NIH K-award grant aimed at testing mechanistically specific and neuroscience-informed interventions for youth AUD.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT SUMMARY/ABSTRACT Post-traumatic stress and alcohol use disorder (PTSD/AUD) are frequently comorbid and are a major US health burden. Individuals with comorbid PTSD/AUD manifest complex symptoms including greater risk for alcohol drinking and relapse, sleep disturbances, hyperarousal, drinking-related aggression, and increased suicidal ideations. Identification of the molecular mechanisms underlying PTSD/AUD disorder will aid the development of novel therapeutic strategies. In this proposal, my focus will be on two key components of the extended amygdala (EA), the central amygdala (CeA) and the bed nucleus stria terminalis (BNST). The CeA and BNST are regions essential for the regulation of alcohol consumption, stress, and anxiety. In the EA, overactive brain stress systems are hallmark feature of comorbid PTSD/AUD, primarily driven by the pro-stress neuropeptide, corticotrophin releasing factor (CRF), resulting in long-lasting negative emotional states. New research has highlighted that the neuropeptide somatostatin (SST) displays anxiolytic (anti-stress) and alcohol-reducing properties and may act to oppose the effects of CRF in the brain. Both CeA and BNST are abundant in SST/CRF, and blockade of CRF or activation of SST system reduces consumption of alcohol and anxiety-like behavior in various rodent models and species. In this proposal, I will apply a multidisciplinary approach (e.g., in situ hybridization, ex vivo electrophysiology, chemogenetics, viral gene transfer of SST, and site-specific behavioral pharmacology) to understand the role of SST system in the EA in PTSD/AUD comorbidity. My overarching hypothesis is that SST signaling may exert reductions of PTSD/AUD phenotypes counteracting an “overactive” CRF in the EA. During my K99 mentoring phase, I will receive critical training in in situ hybridization, and ex vivo electrophysiology to understand neuronal expression and synaptic function of SST/CRF systems in PTSD/AUD. During my independent R00 phase, I will continue to perform electrophysiology, and apply site-specific behavioral pharmacology combined with chemogenetics and viral expression of SST to examine the behavioral modulation of PTSD/AUD by SST/CRF systems. The goal of this MOSAIC K99/R00 is to enhance my training in in situ hybridization and electrophysiological methods to complement my prior training in addiction neuroscience. This award will also provide critical professional career development to guide me in building an independent research program centered around stress disorders and AUD. Collectively, the proposed work will provide novel insight into the mechanistic role of SST and its potential therapeutic use for PTSD/AUD.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Abstract: Alcohol-Associated Liver Disease (ALD) is a major public health problem, accounting for nearly half of all liver-related deaths in the United States. Despite the clear benefits of alcohol abstinence, achieving and maintaining abstinence remains a significant challenge for many ALD patients. Contingency Management (CM), an evidence-based behavioral intervention that provides tangible rewards for positive behavior change, has shown promise in promoting abstinence in alcohol use disorders. However, the application of CM in ALD patients has been limited, partly due to challenges in monitoring alcohol use over extended periods. This study proposes a novel approach that integrates CM with phosphatidylethanol (PEth) testing, an innovative alcohol biomarker with a longer detection window, to promote alcohol abstinence in ALD patients in the hepatology clinic. The specific aims are: 1) to establish the feasibility and acceptability of PEth-based CM for alcohol use in the hepatology clinic for ALD patients; 2) to evaluate the preliminary efficacy of a PEth-based CM intervention in achieving alcohol abstinence; and 3) to explore sociodemographic and clinical factors that are associated with treatment response (alcohol abstinence and alcohol reduction). The study will conduct a pilot randomized controlled trial (REINFORCE Trial) to compare the effectiveness of a 12-week PEth-based CM intervention against a control group receiving treatment as usual in a cohort of ALD patients with alcohol use in the hepatology clinic. The primary outcome will be alcohol abstinence, with secondary outcomes including alcohol reduction, treatment engagement, liver disease severity, and health-related quality of life. Exploratory analyses will examine potential moderators of treatment response. This innovative study addresses a critical gap in the treatment of alcohol use in ALD patients and has the potential to significantly advance scientific knowledge, improve clinical practice, and reduce the public health burden of alcohol-related liver disease. The findings will inform the development of personalized treatment approaches and guide the design of future large-scale clinical trials.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Project Summary/Abstract My professional career goal is to conduct independent research in life sciences defining complex biological systems, including and specifically alcohol-associated liver disease (ALD). ALD is a progressive disease that can lead to irreversible and fatal liver damage in the form of inflammation, fibrosis, cirrhosis, and cancer. Unfortunately, therapeutic treatments for ALD are limited. This is in part due to the lack of understanding of ALD progression at the cellular level. The onset of ALD is represented by a reversible steatotic liver stage where lipid droplets (LDs) excessively accumulate within hepatocytes, the highly metabolizing cell type of the liver. This observation suggests chronic alcohol consumption prevents the breakdown of LDs. Previous studies have shown alcohol disrupts endocytic vesicle trafficking proteins, including Rab5, and LD catabolism; however, these cellular mechanisms of perturbation are poorly understood. Therefore, this proposal aims to elucidate the molecular mechanisms in which alcohol affects Rab5 and endocytic trafficking during ALD progression. Preliminary studies show alcohol disrupts Rab5 subcellular localization without affecting Rab5 activity. Altogether, the central hypothesis is alcohol disrupts Rab5 prenylation, a post-translational modification, to drive its altered localization. This will be tested by two specific aims. Aim 1 will define the impact of alcohol on Rab5 subcellular localization and remodeling of the endosome proteome. Aim 2 will determine the impact of alcohol on global changes in protein prenylation (prenyl-proteome) including Rab5. To accomplish these aims, I will acquire training in confocal microscopy and mass spectrometry. This new training will broaden my current skillsets regarding yeast cell culture and genetics, small molecule trafficking and signaling, the use and design of fluorescent biosensors, and inter-organelle communication. My strong advisory team, including my sponsor and co-sponsor Dr. Micah Schott and Dr. Carol Casey, respectively, one collaborator, and five other faculty have significant expertise in the proposed topics and the desired training objectives. The microscopy and mass spectrometry core facilities, and research environment at UNMC will enhance this research and training. Together, I will have the expertise and training needed to successfully accomplish these aims. The results gained from the proposed research will provide a mechanistic understanding of disrupted endocytic trafficking by alcohol in ALD.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT SUMMARY Background. Firefighters are repeatedly exposed to potentially traumatic events and many experience posttraumatic stress (PTS) symptoms as a result. Over 50% of firefighters report past-month heavy alcohol use and many use alcohol to “self-medicate” from symptoms of hyperarousal, intense negative affect, and insomnia that follow traumatic events. Transcutaneous auricular neurostimulation (tAN) of the vagus nerve has shown promise for alleviating symptoms of PTS and opioid withdrawal symptoms but its effects have yet to be tested in a sample of firefighters with alcohol use disorders (AUD) and co-occurring PTS symptoms. Aims. We will collaborate with a firefighter Community Advisory Board to develop a culturally informed, Phase II randomized clinical trial (RCT) protocol testing tAN as an intervention that may reduce PTS symptoms driving heavy alcohol use for firefighters with AUD (Aim 1; Year 1). We will then conduct a feasibility trial of the protocol (Aim 2; Year 2; N = 20) using the Sparrow device—a patient-administered, ear worn device that delivers electrical stimulation to branches of the vagus and trigeminal nerves—to evaluate the acceptability and feasibility of the protocol, and the appropriateness of the tAN intervention for firefighters with PTS/AUD. Using data from the feasibility study, we will complete an FDA pre-submission with the revised trial protocol to support a PTS/AUD indication for Sparrow Ascent, finalize the protocol with FDA feedback, and write an R01 application for a fully powered Phase II RCT (Aim 3; Year 3). Methods. The feasibility trial will recruit firefighters who engage in heavy drinking, have at least a mild AUD, and report above-average PTS symptoms. Participants will use the Sparrow device for at least 1 hour/day for 5 days/week throughout the 4-week treatment phase, but may use the device more often if they wish. Participants in the active tAN condition (n = 10) will receive therapeutic stimulation from Sparrow; active sham participants (n = 10) will feel stimulation at the trigeminal site only, but below a therapeutic level. They will complete daily measures of traumatic event exposure (number and type), PTS symptom severity, alcohol use (standard drinks) and motivations (trauma-related drinking), and audio journals capturing their experience with symptoms and the device (e.g., helpfulness). Additionally, wearable sensors will record heart rate, heart rate variability, and sleep data throughout the intervention phase. Assessments immediately following treatment (Week 4) and one month later (Week 8) will measure protocol feasibility and tAN’s clinical effects outcomes including insomnia, mental health symptoms, and quality of life. Outcomes. The primary feasibility outcome is the mean of protocol ratings on the Feasibility of Intervention Measure. Secondary feasibility outcomes include ratings of protocol acceptability (Acceptability of Intervention Measure) and Sparrow Ascent appropriateness (Intervention Appropriateness Measure). Exploratory clinical outcomes include reductions in PTS symptom severity scores and heavy drinking events.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT SUMMARY The progression of alcohol use disorder (AUD) is characterized by the emergence of inflexible behaviors, such as compulsive drinking, that exacerbate negative outcomes. Substantial evidence links behavioral inflexibility to hypoactivity of the prefrontal cortex, suggesting that deficient cognitive control over behavior drives compulsive drinking. Despite this, current AUD treatments do not address cognitive dysfunction, leaving inflexible behaviors unchecked. Importantly, chemogenetic activation of the dorsomedial prefrontal cortex prevents the progression of compulsive-like drinking in rats. This identifies a promising new avenue for therapeutic development: prefrontocortical disinhibition. Coupled with existing findings, my preliminary data suggest that negative allosteric modulation of α5-subunit containing GABAA receptors (α5GABAAR) may restore behavioral flexibility through disinhibitory action in the prefrontal cortex. To test this, I will implement an operant reversal-learning task to evaluate goal-directed behavior, a core component of behavioral flexibility (Aim 1), and a quinine-challenge to evaluate inflexible, aversion-resistant drinking (Aim 2). Both aims will utilize systemic administration of an α5GABAAR negative allosteric modulator, to model clinical application, and local infusion to directly interrogate the role of prefrontocortical α5GABAARs. Successful completion of these aims will validate prefrontocortical α5 subunit-containing GABAA receptors as a promising, pro-cognitive drug target addressing inflexible behaviors in alcohol use disorder and other neuropsychiatric conditions characterized by cognitive dysfunction. This project expands upon the PI’s existing expertise to provide training on animal behavior and in vivo pharmacological manipulations, which will optimize technical development as an independent researcher examining the cortical circuits driving behavioral flexibility.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

PROJECT SUMMARY/ABSTRACT Alcohol Use Disorder (AUD) and Post-Traumatic Stress Disorder (PTSD) frequently co-occur, exacerbating symptom severity and complicating treatment outcomes. AUD impairs stress regulation and heightens vulnerability to PTSD, while PTSD promotes maladaptive alcohol consumption as a coping mechanism. Both disorders involve GABAergic and glutamatergic signaling disruptions, resulting in hyperexcitability within the medial prefrontal cortex (mPFC). Acute alcohol exposure enhances inhibitory tone through GABAergic mechanisms, but chronic use reduces inhibition and amplifies excitation, paralleling the elevated excitatory activity seen in PTSD. The dysfunction of somatostatin-expressing interneurons, which maintain inhibitory tone, contributes to this shared pathophysiology. This proposal hypothesizes that stress exposure disrupts excitation-inhibition (E-I) balance in the mPFC driving increased alcohol consumption and heightened motivation for alcohol use, and that targeting metabotropic glutamate receptor 7 (mGluR7), a presynaptic regulator of glutamatergic transmission, can restore this balance. Using single prolonged stress (SPS) to model stress alongside alcohol exposure paradigms such as drinking- in-the-dark (DID) and operant self-administration, I will employ electrophysiology, dual-color fiber photometry, and behavioral assays to quantify E-I balance changes and evaluate the efficacy of AMN082, a mGluR7 allosteric agonist, in reducing pathological alcohol use and PTSD-like behaviors. In Aim 1, I will assess the impact of stress on alcohol consumption in binge drinking. Aim 2 will examine the effects of stress on motivation for alcohol use in mice with prolonged alcohol exposure. Despite known sex differences in AUD neurobiology, the interaction between sex and altered synaptic transmission in comorbid AUD/PTSD remains poorly understood and will be explicitly investigated. Through these aims, I will explore the shared circuit mechanisms between AUD and PTSD and distinct E-I balance differences associated with binge drinking versus long-term alcohol use. This study seeks to identify novel intervention strategies targeting E-I balance, offering dual-benefit approaches to mitigate alcohol use and PTSD symptoms and ultimately improve treatment outcomes. This proposed project will be conducted in the labs of Drs. Jane Taylor and Alicia Che, within Yale University's Departments of Psychiatry and Psychology, and provide me with expertise in electrophysiology, fiber photometry, and behavior, essential for successfully achieving the project's aims. Yale University and the Interdepartmental Neuroscience Program offer a robust scientific community that fosters opportunities for professional development in scientific communication and mentoring. The training and mentorship provided through this fellowship will equip me to become a successful independent researcher focused on understanding the interactions between AUD and PTSD.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Abstract Alcohol abuse is a common and destructive behavioral pattern, accounting for roughly 178,000 deaths per year in 2020 and 2021 (Centers for Disease Control and Prevention, 2024). We know that people generally use alcohol to control physical pain (Brennan et al., 2011; Riley & King, 2009), so much so that pain is understood to be a significant risk factor for alcohol use disorder (e.g., Boissoneault et al., 2019; Jakubczyk et al., 2016; Witkiewitz & Vowles, 2018). We also know that “overdrinking” alcohol is often associated with impulsivity and self-regulatory failure (Bates & Labouvie, 1995; Grau & Ortet, 1999; Hustad et al., 2009). Thus, contextual factors that enhance pain experiences and/or diminish self-regulatory restraint may, in effect, increase one's risk of abusing alcohol. One particular type of social experience—social rejection—may be especially powerful in terms of increasing problem drinking behavior, due to its association with both pain and poorer self-control. Furthermore, people with anxious attachment—a trait distinguished by fear of rejection and a general hypersensitization towards threats (Ainsworth et al., 1978; Bretherton & Parke, 1992)—may be especially vulnerable to the influence of rejection on their alcohol use. Anxious attachment is even considered a significant risk factor for substance use disorders (SUD; e.g., Unterrainer et al., 2017). To date, no research exists on the mechanisms underlying the association between social rejection and “in- the-moment” alcohol use, nor vulnerability factors that enhance the association between rejection and alcohol use. With this application, we propose two studies to explore the link between social rejection and drinking behavior. For Study 1, we will conduct an ecologically valid, longitudinal daily diary study with 188 young adults (aged 21 – 25) to examine how fluctuations in perceptions of rejection are associated with alcohol use, whether attachment anxiety strengthens that association, and whether that association is explained by pain experience and self-regulatory failure. In Study 2, we will recruit 152 carefully screened participants (aged 21 – 25) to participate in a double-blind between-subjects experiment to address the causal nature of the associations expected in Study 1. Findings are expected to contribute to the next step in the research program: developing and testing interventions to reduce impulsive drinking following social rejection.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Sexual and gender minority persons face substantial health inequities, including high rates of unhealthy alcohol use (UAU). High rates of UAU are a consequence of a lack of social safety and social stressors due to systemic oppression that are oftentimes supportive of greater UAU. Despite the important influence of primary partners on UAU and demonstrated efficacy of couples-based treatment for alcohol use disorder in heterosexual couples, the vast majority of alcohol interventions with sexual and gender minority populations have been individually focused. Based on our conceptual and empirical understanding of UAU, we recently completed a rigorous iterative development and pilot testing process for a 3-session couples-based alcohol intervention (called “ReACH2Gether) for cisgender male couples in which at least one partner was receiving care for HIV. In a one-arm pilot trial, the intervention, which was delivered entirely by videoconferencing and allowed for a range of alcohol reduction goals for both members of the couple, was feasible and highly acceptable. Participants showed meaningful and significant changes in alcohol use and key couples-related mechanisms of change. Based on our successful findings, we propose to adapt ReACH2Gether to meet the diverse needs of sexual and gender minority couples and to provide additional content suggested by our pilot work. Guided by a community advisory board (CAB) that represents the diversity of sexual and gender identities who will participate in the project, we will use a two-step sequence of treatment development and evaluation. Stage 1a includes: (1) working with our CAB to adapt our existing couples-based intervention to be inclusive of diverse sexual and gender identities and to incorporate additional content guided by interdependence and social safety theories, (2) conducting a one-arm pilot trial with 12 couples to refine the manual and study procedures, and (3) analyzing exit interviews to finalize refinements to the intervention. Stage 1b consists of a pilot randomized controlled trial with 30 sexual and gender minority adults with UAU and their partners (total N = 60) to test feasibility, acceptability, and potential promise of the intervention. Couples will be randomized to ReACH2Gether or brief advice on alcohol and couples communication (with ReACH2Gether offered after 12 weeks). Alcohol use and mechanisms of behavior change will be assessed at 6-, 12-, and 18-week follow-ups. We will share study results with our CAB and conduct a focus group with key informants, who are involved in delivering behavioral healthcare to sexual gender minority populations, in order to assess the appropriateness and feasibility of deploying ReACH2Gether in their practice contexts. The proposed research plan is well aligned with the prioritization of sexual and gender minority communities for health disparities research and NIH’s strategic plan to advance research on the health of sexual and gender minorities. Findings will provide the necessary groundwork to examine intervention efficacy and implementation processes in a future, large-scale, randomized hybrid effectiveness-implementation trial.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

Alcohol consumption is a major risk factor for cerebrovascular complications and neuroinflammation, yet the underlying mechanism of alcohol-induced cerebrovascular damage remains unclear. Recent studies suggest that lysophosphatidic acid (LPA) plays a role in blood-brain barrier (BBB) dysfunction through Rho kinase signaling. This proposal presents evidence that heavy alcohol consumption (HAC) enhances cerebrovascular autotaxin levels and decreases lipid phosphate phosphatase-3 (LPP3) expression in brain microvascular endothelial cells, leading to increased LPA signaling in the cerebrovasculature. Reactive oxygen species (ROS) increase in the cerebrovasculature following HAC, promoting autotaxin expression through the redox-sensitive transcription factor NFAT and suppressing LPP3 through miR-92a expression. The proposal aims to investigate the role of autotaxin and LPP3 expression in HAC using cellular, mitochondrial, pharmacological, and molecular biological approaches. The proposed studies will test the hypothesis that the LPA axis critically regulates mitochondrial redox balance, endothelial activation, and neuroinflammation via aggravating BBB damage in HAC. The three specific aims include assessing the effect of alcohol on mitochondrial function and redox balance, evaluating the ability of LPP3 to regulate the alcohol-LPA axis on BBB and neuroinflammation, and assessing the neuroprotective effect of autotaxin blockade to mitigate the impact of HAC in the cerebrovasculature. This proposal presents the first evidence that heavy alcohol consumption (HAC) enhances cerebrovascular LPA signaling, leading to neuroinflammation via aggravating BBB damage, and suggests autotaxin inhibition as a potential therapy for alcohol-induced cerebrovascular damage and LPA signaling. The findings of this study could improve our understanding of the mechanism underlying alcohol-induced cerebrovascular injury and provide a foundation for developing targeted therapies to treat these conditions.

NIAAA - National Institute on Alcohol Abuse and Alcoholism

3. Abstract describing research plan for R33 The R33 phase will investigate the mechanistic effects of exogenous ketone supplementation (KS; 25g) on brain nicotinamide adenine dinucleotide (NAD+) concentrations and neurocognitive function during acute alcohol exposure in healthy adult drinkers. Building on the R21 findings demonstrating that 25g KS significantly reduced breath and blood alcohol concentrations and that 7T downfield proton magnetic resonance spectroscopy (¹H- MRS) scans reliably quantify the cerebral NAD+ peak at 9.3 ppm, the R33 will test whether ketone-induced modulation of NAD+-dependent metabolism alters central neurobiological and behavioral responses to alcohol. Forty healthy men and women (ages 21-50), who report at least one recent heavy drinking episode but do not meet criteria for alcohol use disorder, will participate in a randomized, counterbalanced, two-period crossover study. Participants will complete two experimental sessions separated by a washout period. In one session, participants will receive 25g KS; in the other, a taste-matched placebo. Thirty minutes following beverage ingestion, participants will undergo multimodal 7 Tesla MRI scanning, including structural imaging, baseline NAD+ quantification using downfield ¹H-MRS, and functional MRI during a Stop Signal Task (SST) to assess inhibitory control. Alcohol will then be administered outside the scanner to achieve a target breath alcohol concentration (BrAC) of 0.08%. Participants will return immediately to the scanner for repeated NAD+ ¹H-MRS acquisition, SST functional MRI, and exploratory spectroscopy to quantify brain alcohol and additional metabolites (e.g., β-hydroxybutyrate, glutamate). Venous blood samples will be collected to quantify alcohol, NAD+, and ketone concentrations. The primary hypothesis is that KS will increase brain NAD+ concentrations relative to placebo and attenuate alcohol-induced reductions in NAD+ levels. The primary outcome will be the interaction between treatment (KS vs placebo) and alcohol (pre- vs post-alcohol) on NAD+ concentration at the 9.3 ppm peak. Secondary hypotheses are that KS will attenuate alcohol-induced impairments in inhibitory control (indexed by stop-signal reaction time) and modulate blood oxygenation level-dependent (BOLD) activation within fronto-temporal inhibitory control networks. Exploratory analyses will examine associations between blood and brain NAD+ levels, alcohol pharmacokinetics, and cognitive performance, and will assess potential moderation by sex and genetic variation in alcohol-metabolizing enzymes. This study is innovative in combining peripheral pharmacokinetic assessment, high-field NAD+ spectroscopy, and task-based functional neuroimaging within a controlled alcohol challenge paradigm. By directly examining NAD+-dependent mechanisms in vivo, the R33 phase will provide mechanistic insight into how ketone supplementation may alter alcohol metabolism and sensitivity at both peripheral and central levels. Findings will advance understanding of cellular and molecular mechanisms of alcohol tolerance and may identify NAD+- dependent metabolic modulation as a novel target for prevention or intervention strategies for alcohol misuse and alcohol use disorder. 4. Aims page revision The Specific Aims have been modified in a minor way from the original peer-reviewed and approved application. In the originally proposed R33 phase, participants were to receive either 10g or 25g of ketone supplement (KS), with final dose selection contingent upon R21 findings. Based on the R21 results, which demonstrated that 25g KS significantly reduced breath and blood alcohol concentrations (p<0.05) whereas 10g did not produce significant pharmacokinetic effects, the R33 phase will utilize the 25 g dose exclusively. This modification reflects data-driven optimization consistent with the original study design and milestone framework. The overall scientific premise, hypotheses, and mechanistic focus on NAD+-dependent modulation of alcohol sensitivity remain unchanged. No other aims, hypotheses, or outcome measures have been altered.