Pilot Core

Name: Song Young Park, PhD, Associate Professor, School of Health and Kinesiology, UNO

Approved Budget: $25,000

Abstract: Loss of muscle function and reduced exercise tolerance, which are, in part mediated by alterations in the capacity to transport and use oxygen, are two hallmarks of aging. Binge drinking has been increasing among older adults over the last two decades and chronic alcohol misuse can lead to a clinical condition, chronic alcohol myopathy, which is also characterized by a decline in muscle mass, muscle weakness, and increased fatigability. Alcohol misuse may thus exacerbate the age-related decline in oxygen transport and muscle function, and, in turn, accelerates the loss of physical function. Accordingly, the objective of this pilot project is to examine the influence of aging on the acute effects of binge drinking on O2 transport and skeletal muscle function. Our pre-clinical study will test the central hypothesis that age exacerbates the alcohol-mediated impairment of contractile, metabolic, and vascular functions.

Name: Christopher Cummings, MD, PhD, Assistant Professor & Clinical Geneticist, Pediatrics (Genetics)

 Approved Budget: $25,000

 Abstract: Fetal alcohol spectrum disorders (FASD) result from prenatal alcohol exposure and are a common cause of developmental and behavioral abnormalities. However, due to an incomplete understanding of the mechanism, targeted therapies are unavailable currently. DNA methylation is known to be disrupted in patients with FASD, but other related aspects of the epigenetic regulatory network remain unexplored. In this study, we propose to study chromatin accessibility, a separate but related epigenetic feature to DNA methylation, in neurologically relevant model systems, namely a neural progenitor cell line and a mouse model of FASD. We will also profile gene expression differences in these systems, to tie abnormalities in chromatin regulation to alterations in underlying gene expression. Together, these studies will provide a more comprehensive understanding of the mechanism of alcohol on the developing neural system, and aid in the identification of potential therapeutic targets through future studies supported by this study’s data.

Name: Siddappa Byrareddy, PhD, Vice Chair for Research, Professor, PEN

Approved Budget: $25,000

 Abstract: The combination of alcohol use and HIV infection has been shown to negatively impact the immune system, potentiallyhindering the effectiveness of currently available antiretroviral therapies. Understanding the interplay between alcohol use and HIV reservoirs is a critical area of study that can inform better treatment strategies for people living with HIV (PWH) suffering from alcohol use disorders (AUD). There is knowledge of gaps, and it has been hypothesizing that alcohol abuse may influence the size of viral reservoirs in various tissues and organs, particularly within the central nervous system (CNS).

The above observations are significant because the penetration of antiretrovirals in the CNS is often poor, leading to increased viral replication and the establishment of reservoirs. To investigate these differences, we propose to utilize cryoarchived samples from a well-established rhesus macaque model of alcohol dependence. This approach will allow us to delineate the impact of alcohol on viral reservoirs across diverse tissues and organs. The data obtained from this study will serve as proof of principle regarding the influence of alcohol on viral reservoirs. Additionally, it will generate preliminary data necessary for an R01 proposal to the NIH aimed at a comprehensive understanding of AUD-mediated immune alterations in the context of HIV cure research efforts.

Name: Sundararajan Mahalingam, PhD, Instructor, Department of Biochemistry and Molecular Biology

 Approved Budget: $25,000

 Abstract: Our previous data has shown that chronic binge alcohol exposure to mice decreases hepatic lysosome-associated membrane protein type 2a (LAMP2A) content, indicating dysregulation of chaperone-mediated autophagy (CMA). CMA-promoting pharmacological agents can maintain/restore CMA activity in ethanol-exposed liver cells and reduce LD accumulation. Selectively activating CMA with pharmacological agents like atypical retinoid 7 (AR7), guanidine retinoid 2 (GR2), QX77, CA77.1, and CA 39, have been shown to be good pharmacological CMA activators in different models. Preliminary studies with AR7 shows promising results in alleviating EtOH-induced fatty liver by increasing CMA-dependent LD breakdown, via increasing LAMP2A expression. However, few reports have been suggested that AR7 is not suitable for in vivo applications. Here we propose using QX77, a novel potent CMA activator derived from AR7 that activates CMA by antagonizing retinoic acid receptor-a (RARa) signaling through its binding to ligand-binding domain and triggers the expression of key CMA components. In addition, QX77 induces the upregulation of LAMP2A (a rat limiting protein in CMA) and Rab11 (facilitate LAMP2A trafficking to lysosome) expressions. Therefore, here, we propose to test the potential of a novel CMA activator, QX77, in enhancing chaperone-mediated autophagy to alleviate ethanol-induced fatty liver, both in vitro and in vivo experimental conditions.

View the latest RFA

Examples of pilot or feasibility projects include:

• Initial support for developing novel, innovative, cutting-edge research, translation, prevention, or intervention approaches.
• Adapting and evaluating proven research tools or techniques for new applications or delivery systems.
• Obtaining preliminary data, mining existing state or national datasets, or pursuing critical data gaps.
• Supporting investigators from other fields of study to apply their expertise to alcohol use/misuse challenges.
• Developing new mechanisms for external or multicenter collaborative partnerships to address emerging alcohol use/misuse concerns.
• Exploring new directions that represent significant departures from established approaches, yet have the potential to yield great impacts.

Name: Nicole Zabik, PhD, Post-Doc Research Associate, MMI Administration Research

Approved Budget: $25,000

Abstract: Alcohol use disorders (AUDs) are a debilitating condition that affect 30% of Americans. Relapse is common – making long-term recovery difficult. A primary reason for high relapse rates is the development of anxiety during early abstinence that triggers alcohol use. That is, individuals engage in negative reinforcement by using alcohol to relieve anxiety that develops during abstinence. The ‘alcohol exposome’ is also linked to relapse, likely due to its influence on negative reinforcement behaviors. Negative reinforcement and the alcohol exposome contribute to mechanisms of recovery and relapse, underscoring their importance in behavioral and neural mechanisms of AUD. To address this, we translated a well-validated animal model that measures individual differences in negative reinforcement to determine (1) negative reinforcement neural circuits and behaviors and (2) the impact of the alcohol exposome on negative reinforcement. Defining these mechanisms is a crucial step towards identifying novel targets for preventing relapse in AUDs.

Name: Armen, Petrosyan, MD, PhD, Associate Professor, Department of Biochemistry and Molecular Biology

Approved Budget: $25,000

Abstract: Radiation therapy (RT) is integral in treating prostate cancer (PCa), but challenges persist, particularly in pelvic regions where unintentional radiation induces significant fibrosis in the surrounding organs and tissues. PCa survivors often exhibit increased alcohol consumption, impacting RT effectiveness; however, the mechanism is enigmatic. Both RT and alcohol cause scattering of Golgi, stress of the endoplasmic reticulum (ER), and activation of the ATF6 branch of ER stress response (ERSR). This proposal studies the impact of alcohol and irradiation co-treatment on secretion of adiponectin (APN), the important glycoprotein that is secreted by primary mouse adipocytes and protects fibroblasts from radiation-induced cell death, myofibroblast formation, and senescence. We hypothesize that alcohol treatment significantly accelerates the damaging effect of radiation on Golgi, leading to a) alteration of APN glycosylation and secretion in adipocytes and b) accelerated autophagy in fibroblasts followed by activation of ATF6-mediate ERSR. Ultimately, this accelerates transformation of fibroblasts into myofibroblasts.

Name: Mathilda Willoughby, PhD, Post-Doc Research Associate, Department of Biochemistry and Molecular Biology

Approved Budget: $25,000

Abstract: Alcohol-associated liver disease (ALD) is the leading cause of alcohol related mortality, which strongly supports an urgent need to fully characterize and prevent irreversible liver damage. Chronic ethanol (EtOH) consumption leads to steatosis, an accumulation of lipid droplets (LDs). The over-accumulation of LDs plays a principal role in progression of ALD by promoting inflammation, cirrhosis, and cancer. However, the molecular mechanisms by which EtOH disrupts LD catabolism are not fully understood. Fortunately, EtOH has been shown to disrupt endocytic trafficking, a pathway that closely overlaps with lipid catabolism. The current proposed studies will endeavor to more fully understand how chronic alcohol consumption affects endocytic trafficking and the potential consequences related to LD catabolism in hepatocytes. Specifically, we will investigate the endocytic protein, Rab5, under the influence of cellular EtOH, whereby we will map its subcellular distribution and characterize the regulation of Rab5 post-translational modifications. These studies will provide preliminary data for future grant applications (F32) to develop novel target therapeutic interventions for ALD.

Name: Seema Singh, PhD, Instructor, Pharmacology and Exp Neuroscience

Approved Budget: $25,000

Abstract: Almost half of the people living with HIV (PLWH) are alcohol abusers and have a higher prevalence (2–3-fold) of developing alcohol use disorders (AUD), which remains a major public health concern. This pilot study aims to investigate the molecular mechanisms underlying neuroinflammation in the context of AUD in HIV-infected individuals. We recently identified that astrocyte-specific inflammasome NLRP6 (NOD-like receptor family pyrin domain containing 6) protein plays a critical role in HIV Tat and substance misuse-mediated neuroinflammation and is epigenetically regulated by miRNA-339. Based on the premise that alcohol use exacerbates HIV- associated neuroinflammation, we aim to explore the epigenetic regulation of NLPR6-mediated neuroinflammation in HIV Tat and ethanol-exposed astrocytes. The findings of this study could have important implications for the future development of therapeutic interventions aimed at mitigating neuroinflammation in AUD in HIV-1 infected individuals and, thus, relate to the ACORN P50 grant mission.

View the latest RFA

Examples of pilot or feasibility projects include:

• Initial support for developing novel, innovative, cutting-edge research, translation, prevention, or intervention approaches.
• Adapting and evaluating proven research tools or techniques for new applications or delivery systems.
• Obtaining preliminary data, mining existing state or national datasets, or pursuing critical data gaps.
• Supporting investigators from other fields of study to apply their expertise to alcohol use/misuse challenges.
• Developing new mechanisms for external or multicenter collaborative partnerships to address emerging alcohol use/misuse concerns.
• Exploring new directions that represent significant departures from established approaches, yet have the potential to yield great impacts.

Micah Schott, PhD, Assistant Professor, Department of Biochemistry and Molecular Biology

Approved Budget (7-1-23 to 6-30-24): $25,000

Abstract: Alcohol-associated liver disease (ALD) is a growing public health concern, and new therapeutic strategies are urgently needed to prevent its progression. This proposal establishes a collaboration between the laboratories of Dr. Micah Schott and co-investigator Dr. Rebecca Oberley-Deegan to investigate cellular mechanisms underlying ALD and explore the potential of antioxidant therapy using the ROS-scavenging agent BuOE to mitigate liver damage. The accumulation of lipid droplets (LDs) in hepatocytes plays a central role in ALD progression, and the role of LDs in facilitating hepatocyte damage, specifically through lipid peroxidation, is poorly understood. This project aims to define the impact of lipolysis on lipid peroxidation and cell damage in hepatocytes exposed to EtOH. Furthermore, the efficacy of BuOE, a promising SOD-mimic antioxidant, in reducing ROS-driven hepatocyte damage in ALD will be investigated using in vitro cell models and in vivo mouse models that recapitulate key features of human ALD. This study will provide preliminary data for future grant proposals to develop novel therapeutic interventions for ALD.

Shibiao Wan, PhD, Assistant Professor, Department of Genetics, Cell Biology and Anatomy

Approved Budget (7-1-23 to 6-30-24): $25,000

Abstract: Alcohol use disorder (AUD) is a type of addiction with adverse health, occupational, and social consequences. Existing neuroimaging studies indicate that AUD is a chronic relapsing brain disease, and it is related to aberrant functional connectivity (FC) in a unifying triple network. However, these studies have following limitations: (1) the results might be statistically unreliable due to the insufficiency of multiple-comparison correction when dealing with millions of dimensions of brain networks; (2) the biomarkers studied were all within triple networks, whereas few attentions were paid to other brain networks; and (3) complex patterns in a multi-variate matter could not be detected. To address these concerns, we propose to develop a machine learning (ML) framework to automatically identify FC biomarkers from both within- and between-brain networks (including both the triple network and other brain networks) for AUD diagnosis. Our proposed ML model is extensible to study other addictions and neurological disorders.

Siva S Koganti, PhD, Instructor, Department of Internal Medicine, Gastroenterology & Hepatology Division

Approved Budget (7-1-23 to 6-30-24): $25,000

Abstract: About 48% of HIV-infected individuals are alcohol abusers. Alcohol exposure tremendously potentiates HIV-induced liver fibrosis. The mechanisms behind these events are unclear. Emerging pieces of evidence point to the role of microRNAs (miRs) in EVs as the regulators of hepatic stellate cell (HSC) activation. We recently demonstrated that differentially expressed miRs in extracellular vesicles (EVs) released from alcohol-treated HIV-infected hepatocytes target mRNAs in HSC to regulate the activation of pro-fibrotic genes and liver fibrosis progression. In this context, we propose to functionally validate differentially expressed miRs and to target specific miRs with anti-miR treatment to attenuate fibrogenesis in alcohol-related and HIV-induced liver disease, which improves the quality of treatment and longevity in people living with HIV1 (PLWH) also having Alcohol Use Disorder (AUD) This illustrates the mechanism by which double-exposure to the virus (HIV) and ethanol regulates liver fibrosis progression. Thus, this study fully corresponds to the ACORN P50 grant mission.

Daniel R. Anderson, MD, Associate professor, Department of Internal Medicine- Cardiology

 Approved Budget (10-1-23 to 6-30-24): $20,000

 Abstract: Cardiovascular disease (CVD) is the leading cause of death worldwide. Recently, it has been demonstrated that ethanol consumption is an independent risk factor for the development of many forms of CVD. Unfortunately, the mechanisms by which ethanol causes CVD are not well understood. Our work will determine the electrophysiologic, biochemical, and molecular mechanisms by which EtOH and its metabolic by-products, acetaldehyde and malondialdehyde, affect cardiomyocytes. We hypothesize that ethanol and its oxidative metabolites disrupt the normal function and electrophysiology of cardiomyocytes, predisposing individuals to cardiovascular disease. To test this hypothesis, we propose two aims: Aim 1. Determine the effects of ethanol and its metabolites on the activity of cardiomyocytes. Aim 2. Determine the mechanisms by which by-products of ethanol metabolism alter cardiomyocyte function. Using human inducible pluripotent stem cells differentiated into beating cardiomyocytes, we will determine the effects of treatment with ethanol, acetaldehyde, and malondialdehyde on cardiomyocytes.

Amar B. Singh, PhD, Professor, Department of Biochemistry and Molecular Biology

 Approved Budget (10-1-23 to 6-30-24): $20,000

Abstract: Alcoholic Pancreatitis (AP) is a potentially fatal inflammation of the pancreas; however, molecular understanding of its pathobiology is poor, which has a strong bearing on the clinical management. Dysregulated barrier function promotes inflammation. Tight junctions (TJs) help regulate the barrier function. The claudin family of proteins constitutes TJs where expression of claudin-2 increases during intestinal inflammation. Remarkably, claudin-2 is mutated in patients with AP and associates with an increased risk of AP. However, role of claudin-2 in AP is not known. In preliminary studies using mouse model of AP, we found significant upregulation in claudin-2 expression. This increase matched with simultaneous increases in expression of phospho-NFkB and phospho-Stat3. Based on these findings and the known association of claudin-2 with inflammation, we propose a causal role for claudin-2 in promoting AP, which will be tested in current proposal. We anticipate the outcome from proposed studies to be novel and clinically relevant.