Myogenetic oligodeoxynucleotide (myoDN) suppresses proliferation and induces differentiation of vascular smooth muscle cells

Myogenetic oligodeoxynucleotide (myoDN) suppresses proliferation and induces differentiation of vascular smooth muscle cells

Mana Miyoshi1, Yuma Nihashi2, Koji Umezawa1,3, Takeshi Shimosato1,2,3, Tomohide Takaya1,2,3.

  1. Faculty of Agriculture, Shinshu University.
  2. Graduate School of Medicine, Science and Technology, Shinshu University.
  3. Institute for Biomedical Sciences, Shinshu University.

日本農芸化学会2021年度大会 (仙台), 2021/03/20 (口演).


Objective: Arteriosclerosis is a risk factor for cerebral and cardiovascular diseases that are the major causes of death in Japan. At atheroma of atherosclerosis, vascular smooth muscle cells (VSMCs) de-differentiate, migrate into intima, and abnormally proliferate to induce intimal thickening and vascular stenosis. In Moenckeberg arteriosclerosis, VSMCs calcify with a similar process to bone differentiation, which facilitates rupture of vascular structure. Therefore, regulation of VSMC differentiation is an important subject for prevention and therapy for arteriosclerosis. We recently identified the 18-base myogenetic oligodeoxynucleotide (myoDN), iSN04, that promotes skeletal and cardiac muscle differentiation by targeting nuclear phosphoprotein, nucleolin. In this study, we investigated the effect of iSN04 during VSMC differentiation.

Methods: Commercial human aortic SMCs (hAoSMCs) were cultured with 30 uM iSN04. Cell proliferation was examined by EdU staining and differentiation was investigated by qPCR. To study the effect of iSN04 on bone differentiation, murine osteoblast cell line MC3T3-E1 was utilized. The MC3T3-E1 cells induced differentiation with 10 or 30 uM iSN04 were subjected to alkaline phosphatase (ALP) staining to quantify bone differentiation.

Results: EdU staining showed that the ratio of proliferating hAoSMCs were significantly reduced by iSN04 treatment. qPCR results indicate that expression levels of α-actinin (ACTA2), SM22-α (TAGLN), and caldesmon (CALD1) that are differentiation markers for hAoSMCs were significantly upregulated by iSN04. ALP activities of MC3T3-E1 cells were dramatically decreased by iSN04, suggesting the inhibited bone differentiation.

Conclusion: The present study indicated that iSN04 suppresses proliferation and facilitates differentiation of VSMCs not only of skeletal muscle cells. In addition, iSN04 is expected to inhibit calcification of VSMCs because iSN04 downregulated bone differentiation of osteoblasts. iSN04 can be the nucleic acid drug targeting VSMCs to abrogate hyperproliferation in atherosclerosis and calcification in Moenckeberg arteriosclerosis.

Keywords: arteriosclerosis, myogenetic oligodeoxynucleotide, vascular smooth muscle cell