Dedifferentiation of smooth muscle cells in intracranial aneurysms and its potential contribution to the pathogenesis

Mieko Oka,Isao Ono,Takakazu Kawamata,Hirohiko Imai,Satoshi Shimo,Yu Abekura,Kazuhiko Nozaki,Nobuhiko Ohno,Mika Kushamae,Hirokazu Koseki,Akitsugu Kawashima,Kampei Shimizu,Haruka Miyata,Tomohiro Aoki

doi:10.1038/s41598-020-65361-x

Abstract

Smooth muscle cells (SMCs) are the major type of cells constituting arterial walls and play a role to maintain stiffness via producing extracellular matrix. Here, the loss and degenerative changes of SMCs become the major histopathological features of an intracranial aneurysm (IA), a major cause of subarachnoid hemorrhage. Considering the important role of SMCs and the loss of this type of cells in IA lesions, we in the present study subjected rats to IA models and examined how SMCs behave during disease progression. We found that, at the neck portion of IAs, SMCs accumulated underneath the internal elastic lamina according to disease progression and formed the intimal hyperplasia. As these SMCs were positive for a dedifferentiation marker, myosin heavy chain 10, and contained abundant mitochondria and rough endoplasmic reticulum, SMCs at the intimal hyperplasia were dedifferentiated and activated. Furthermore, dedifferentiated SMCs expressed some pro-inflammatory factors, suggesting the role in the formation of inflammatory microenvironment to promote the disease. Intriguingly, some SMCs at the intimal hyperplasia were positive for CD68 and contained lipid depositions, indicating similarity with atherosclerosis. We next examined a potential factor mediating dedifferentiation and recruitment of SMCs. Platelet derived growth factor (PDGF)-BB was expressed in endothelial cells at the neck portion of lesions where high wall shear stress (WSS) was loaded. PDGF-BB facilitated migration of SMCs across matrigel-coated pores in a transwell system, promoted dedifferentiation of SMCs and induced expression of pro-inflammatory genes in these cells in vitro. Because, in a stenosis model of rats, PDGF-BB expression was expressed in endothelial cells loaded in high WSS regions, and SMCs present nearby were dedifferentiated, hence a correlation existed between high WSS, PDGFB and dedifferentiation in vivo. In conclusion, dedifferentiated SMCs presumably by PDGF-BB produced from high WSS-loaded endothelial cells accumulate in the intimal hyperplasia to form inflammatory microenvironment leading to the progression of the disease.

Highlights

Smooth muscle cells (SMCs) are the major type of cells constituting arterial walls and play a role to maintain stiffness via producing extracellular matrix
Macrophages and endothelial cells were immunonegative for smooth muscle α-actin (SMA), and SMA-immunopositive smooth muscle cells (SMCs) in tunica media were not stained in immunocontrol samples without 1st antibody (Figure S3)
Almost all SMCs in the intimal hyperplasia contained a large amount of mitochondria or rough endoplasmic reticulum (rER) and its number was significantly larger than that in SMCs present in the media of intracranial arteries (Median; 0.2 × 106, 0.8 × 106 in Fig. 2C left panel; 2.1%, 14.7% in Fig. 2C right panel) (Fig. 2B,C, Figure S4), suggesting the activation of SMCs at the intimal hyperplasia

Summary

Introduction

Smooth muscle cells (SMCs) are the major type of cells constituting arterial walls and play a role to maintain stiffness via producing extracellular matrix. At the neck portion of IAs, SMCs accumulated underneath the internal elastic lamina according to disease progression and formed the intimal hyperplasia. Dedifferentiated SMCs presumably by PDGF-BB produced from high WSS-loaded endothelial cells accumulate in the intimal hyperplasia to form inflammatory microenvironment leading to the progression of the disease. We examined the change of SMCs over time and the contribution to the pathological condition during the progression of IA lesions using a rat model

Methods

Results

Discussion

Conclusion