Wednesday, January 15, 2025
1/15/2025
Project Summary Alcohol use disorder (AUD) is a prevalent psychiatric condition in the United States, imposing a huge cost on society and contributing to thousands of deaths each year. While many of the behavioral changes associated with the disorder have been characterized, the underlying physiological conditions, such as neuroinflammation and transcriptomic alterations, require further study. This proposal presents a novel hypothesis for testing the potential mechanism(s) coordinating these molecular and behavioral changes. Integrating previous literature, I hypothesize that the long non-coding RNA (lncRNA) Malat1 may mediate neuroinflammatory responses to ethanol and ethanol consumption. Non-coding RNAs are continually being recognized as integral players in several basic cellular functions. Malat1 has been shown to coordinate transcriptional regulation of gene expression and inflammatory processes, with multiple studies suggesting a link between Malat1 and the high mobility group box 1 (Hmgb1)/Toll-like receptor 4 (Tlr4) neuroinflammatory cascade. In this research proposal, I want to determine if Malat1 regulates expression of the Hmgb1/Tlr4 pathway in response to ethanol, particularly in astrocytes, a CNS cell-type that regulates neuroimmune responses and plays a pivotal role in controlling behavior. The first aim of this proposal will utilize CRISPR/Cas9 to knockdown Malat1 expression in primary astrocyte cultures and assess ethanol-induced changes in neuroimmune gene expression and the production of cytokines. The second aim will create a novel floxed Malat1 conditional knockout mouse which will be bred to a line of mice expressing the recombinase enzyme Cre selectively in astrocytes. Offspring of this mating will possess astrocyte-specific knockout of Malat1. Assessing the ethanol-induced neuroimmune response and ethanol drinking behavior of these animals will provide insight into the role of both Malat1 and astrocytes in regulating the underlying pathology of AUD. This project is an important step in understanding the functional contribution of the lncRNA Malat1 in AUD and how some of the physiological phenotypes of the disorder may be perpetuated. With few effective treatments currently available for individuals suffering from AUD, results obtained from the proposed studies may point to new prevention strategies and therapeutic targets. Additionally, this project will greatly enhance my graduate training, as it will give me the opportunity to design and perform hypothesis driven research in an exciting new area of research and master innovative techniques. Ultimately, the training provided by this Ruth L. Kirschstein National Research Service Award (F31) will aid in my scientific training as a graduate student, as well as in my professional development as an independent investigator.
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