Date of Award
12-18-2024
Degree Type
Thesis
Degree Name
Master of Interdisciplinary Studies (MIS)
Department
Biomedical Sciences
First Advisor
Chunying Li
Second Advisor
Ping Song
Abstract
Introduction: Pathogenic transformation of vascular smooth muscle cells (VSMCs) into osteochondrogenic phenotypes is a key feature of advanced atherosclerotic cardiovascular disease and is driven by transcription factors, like SOX9 and RUNX2. Here, we explore the previously uncharacterized role of SMYD2, a histone lysine methyltransferase, in VSMC phenotypic switching during atherosclerotic plaque formation.
Methods: C3H/10T1/2 (10T1/2) cells, wildtype (WT) and Smyd2-depleted (Smyd2KO), were cultured and treated with 5 mmol/L inorganic phosphate (Pi) for 24 or 72 hours, followed by Western blot (WB) analysis. For RUNX2 inhibition, 10T1/2 cells were treated with CADD522 and 5 mmol/L Pi for 8, 24, 48, or 72 hours, with protein expression analyzed by WB. Co-immunoprecipitation (Co-IP) was performed on WT or Smyd2 KO 10T1/2 cells. Immunoprecipitation was conducted using an antibody against RUNX2, followed by immunoblotting with a SUMO-2/3 antibody to assess RUNX2 SUMOylation. For SUMO inhibition, 10T1/2 cells were treated with ginkgolic acid and 5 mmol/L Pi for 24 hours, with protein expression analyzed by WB.
Results: Western blotting revealed lower RUNX2 protein levels in Smyd2KO 10T1/2 cells. Co-IP demonstrated increased SUMOylation of RUNX2 in Smyd2KO cells compared to wild-type cells. RUNX2 inhibition with CADD522 reduced SMYD2, while SUMOylation inhibition with ginkgolic acid had no effect on SMYD2 and RUNX2 expression.
Conclusion: Smyd2 promotes atherosclerotic plaque formation by regulating SUMOylation of essential transcription factors involved in VSMC osteochondrogenic phenotypic switching. Targeting SMC-specific Smyd2 and SUMOylation pathways may offer a novel therapeutic strategy in preventing vascular calcification in atherosclerosis.
Recommended Citation
Ilse, Nadia M., "SMYD2 Promotes Atherosclerotic Plaque Calcification by Regulating SUMOylation in Vascular Smooth Muscle Cells." Thesis, Georgia State University, 2024.
https://scholarworks.gsu.edu/biomedical_theses/16