Presentation Date
13-10-2022 12:00 AM
Description
Background: With recent advances in antiretroviral therapy, people living with HIV (PLWH) now have a near-normal life expectancy. As such, PLWH experience aging-related comorbidities, such as metabolic disorders and frailty earlier in life than the general population. At-risk alcohol use is twice as likely in PLWH compared to the general population and alcoholic myopathy occurs in 40- 60% of people with an alcohol use disorder. Previous studies demonstrate that chronic binge alcohol (CBA) in simian immunodeficiency virus (SIV)-infected rhesus macaques causes decreased myoblast differentiation and downregulation of microRNA-206 (miR-206) in skeletal muscle. Additionally, lower miR-206 expression is associated with increased expression of myocyte enhancing factor 2C (MEF2C), a transcription factor important for differentiation. Hypothesis: We hypothesize that increasing miR-206 expression in CBA-derived primary macaque myoblasts will increase differentiation. Methods: Eight SIV-infected female rhesus macaques received either water (VEH) or CBA (13- 14 g/kg/week) for 14.5 months. Three months into either VEH or CBA treatment, animals were vaginally infected with SIVmac251, and antiretroviral therapy was started 2.5 months later. Animals were sacrificed after 9 months, and primary cells were derived from vastus lateralis skeletal muscle tissue 24 hours after the last dose of alcohol (blood alcohol=0 mM). Myoblasts at passage 4 (VEH & CBA) were proliferated for three days. Transfection with either miR-206 or miR scramble as a control occurred on day 0 of differentiation creating four groups: VEH+scramble, CBA+scramble, VEH+miR-206, and CBA+miR-206. Cells were allowed to differentiate for five days before analysis. Expression levels of miR-206 were determined using qRT-PCR. Fusion index was calculated by determining the percentage of myonuclei that have fused into myotubes after HEMA 3 staining. Results: Two-way ANOVA revealed increased miR-206 levels (about 150 fold, p
Recommended Citation
Bergeaux, Peter; Bourgeois, Brianna; Molina, Patricia E.; and Simon, Liz, "Chronic Binge Alcohol Impairs Myoblast Differentiation: Role of microRNA-206" (2022). Medical Student Research Poster Symposium. 89.
https://digitalscholar.lsuhsc.edu/sommrd/2022MRD/Posters/89
Included in
Chronic Binge Alcohol Impairs Myoblast Differentiation: Role of microRNA-206
Background: With recent advances in antiretroviral therapy, people living with HIV (PLWH) now have a near-normal life expectancy. As such, PLWH experience aging-related comorbidities, such as metabolic disorders and frailty earlier in life than the general population. At-risk alcohol use is twice as likely in PLWH compared to the general population and alcoholic myopathy occurs in 40- 60% of people with an alcohol use disorder. Previous studies demonstrate that chronic binge alcohol (CBA) in simian immunodeficiency virus (SIV)-infected rhesus macaques causes decreased myoblast differentiation and downregulation of microRNA-206 (miR-206) in skeletal muscle. Additionally, lower miR-206 expression is associated with increased expression of myocyte enhancing factor 2C (MEF2C), a transcription factor important for differentiation. Hypothesis: We hypothesize that increasing miR-206 expression in CBA-derived primary macaque myoblasts will increase differentiation. Methods: Eight SIV-infected female rhesus macaques received either water (VEH) or CBA (13- 14 g/kg/week) for 14.5 months. Three months into either VEH or CBA treatment, animals were vaginally infected with SIVmac251, and antiretroviral therapy was started 2.5 months later. Animals were sacrificed after 9 months, and primary cells were derived from vastus lateralis skeletal muscle tissue 24 hours after the last dose of alcohol (blood alcohol=0 mM). Myoblasts at passage 4 (VEH & CBA) were proliferated for three days. Transfection with either miR-206 or miR scramble as a control occurred on day 0 of differentiation creating four groups: VEH+scramble, CBA+scramble, VEH+miR-206, and CBA+miR-206. Cells were allowed to differentiate for five days before analysis. Expression levels of miR-206 were determined using qRT-PCR. Fusion index was calculated by determining the percentage of myonuclei that have fused into myotubes after HEMA 3 staining. Results: Two-way ANOVA revealed increased miR-206 levels (about 150 fold, p
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