Introduction: Electronic cigarettes pose a serious emerging threat to cardiovascular health. The use of electronic nicotine delivery systems (ENDS) such as electronic cigarettes (e-cigs) have increased exponentially, especially among adolescents. Emerging data suggest e-cig use has been implicated in promoting atherosclerosis. This work explores the mechanism of e-cig exposure accelerated the atherosclerotic lesion development and vascular remodeling. Methods: SMC-lineage tracing mice (n=16) on a ApoE null hyperlipidemic background were put on high fat diet for 12 weeks and exposed to pod-based e-cigs (Juul) three times a week for 12 weeks in a whole-body chamber (inExpose, Scireq) and compared to WT mice exposed to air (n=16). The aortic sinus was dissected and digested, FACS was performed to sort for live single cells, then single-cell RNA-Seq (sc-RNAseq) and ATAC-seq were performed on the 10X Genomics platform. Analyses were completed with Seurat and Signac tools. The osteogenic load was assessed by histology with alkaline phosphatase (AP) enzymatic assay using the Ferangi Blue Chromogen kit. Results: scRNA-Seq analysis revealed that e-cig accelerated the SMC transition to a chondrogenic phenotype (CMC). We found a significantly increased proportion of chondromyocytes (CMC) expressing specific markers such as Col2a1 and Acan in the e-cig exposed mice compared to control. The scATAC-seq profiles showed that e-cig exposed mice develop a cell cluster that enriched for markers related to nicotine, glutamatergic and dopaminergic pathways. The genes highly enriched for chromatin accessibility in this cluster included Grin2a , a gene that encodes a N-methyl-D-aspartate receptor (NMDAR) GluN2A, which was found to be expressed in the fibromyocyte and CMC population of the scRNA-seq. Consistent with our finding from the scRNA-Seq data showing a greater CMC proportion in the e-cig exposed mice, we found the relative AP-stained area to be larger in the exposed group than in the WT group. Conclusion: Overall, these data indicate that e-cig exposure can promote atherosclerosis through adverse SMC phenotypic modulation. Moreover, e-cig exposure induces NMDAR activation that may be implicated in SMC modulation to a chondrogenic phenotype.