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RNA Expression Profiles of Vascular Smooth Muscle Cells Isolated from Mice with and without Bmal1 Deletion

ABSTRACT: To investigate the transcriptional changes in vascular smooth muscle cells (VSMC) induced by diabetic condition medium via Bmal1, we isolated VSMCs from three pairs of Bmal1 flox/flox (wild-type, WT) and Bmal1 Δ/Δ (Bmal1 knockout) littermates. The cells were cultured in diabetic condition medium for seven days. Total RNA was extracted using the RNeasy Mini Plus Kit (Qiagen) and analyzed via RNA sequencing (RNA-seq) to profile transcriptomic changes.

ORGANISM(S): Mus musculus

PROVIDER: GSE290156 | GEO | 2025/02/25

REPOSITORIES: GEO

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Project description:Background: Ectopic vascular calcifications represent a major clinical problem associated with cardiovascular disease and mortality. However, the mechanisms underlying pathological vascular calcifications are largely unknown hampering the development of therapies to tackle this life threatening medical condition. Results: In order to gain insight into the genes and mechanisms driving this pathological calcification process we analyzed the transcriptional profile of calcifying vascular smooth muscle cells (C-VSMCs). These profiles were compared to differentiating osteoblasts, cells that constitute their physiological calcification counterparts in the body. Overall the transcriptional program of C-VSMC and osteoblasts did not overlap. Several genes, some of them relevant for bone formation, were distinctly modulated by C-VSMCs which did not necessarily lose their smooth muscle cell markers while calcifying. Bioinformatics gene clustering and correlation analysis disclosed limited bone-related mechanisms being shared by two cell types. Extracellular matrix (ECM) and biomineralization genes represented common denominators between pathological vascular and physiological bone calcifications. These genes constitute the strongest link between these cells and represent potential drivers for their shared end-point phenotype. Conclusions: the analyses support the hypothesis that VSMC trans-differentiate into C-VSMCs keeping their own identity while using mechanisms that osteoblasts use to mineralize. The data provide novel insights into groups of genes and biological processes shared in MSC and VSMC osteogenic differentiation. The distinct gene regulation between C-VSMC and osteoblasts might hold clues to find cell-specific pathway modulations, opening the possibility to tackle undesired vascular calcifications without disturbing physiologic bone formation and vice versa. Total RNA obtained from hMSC and hVSMC cultured in osteogenic differentiation medium supplemented with 1.8 mM Ca2+ for 0, 2, 8, 12 or 25 days respectively. For each timepoint 3 replicates were used, with exception for day 0 where 4 replicates were collected.

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RNA Expression Profiles of Vascular Smooth Muscle Cells Isolated from Mice with and without Bmal1 Deletion

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