Dysregulation of Cells Cycle and Apoptosis in Human Induced Pluripotent Stem Cells Chondrocytes Through p53 Pathway by HT-2 Toxin: An in vitro Study.

Yanan Zhang,Xiong Guo,Mikko J Lammi,Sijia Tan,Feng’E Zhang,Huan Liu,Xialu Lin,Peilin Meng

doi:10.3389/fgene.2021.677723

Abstract

Kashin–Beck disease (KBD) mainly damages growth plate of adolescents and is susceptible to both gene and gene–environmental risk factors. HT-2 toxin, which is a primary metabolite of T-2 toxin, was regarded as one of the environmental risk factors of KBD. We used successfully generated KBD human induced pluripotent stem cells (hiPSCs) and control hiPSCs, which carry different genetic information. They have potential significance in exploring the effects of HT-2 toxin on hiPSC chondrocytes and interactive genes with HT-2 toxin for the purpose of providing a cellular disease model for KBD. In this study, we gave HT-2 toxin treatment to differentiating hiPSC chondrocytes in order to investigate the different responses of KBD hiPSC chondrocytes and control hiPSC chondrocytes to HT-2 toxin. The morphology of HT-2 toxin-treated hiPSC chondrocytes investigated by transmission electron microscope clearly showed that the ultrastructure of organelles was damaged and type II collagen expression in hiPSC chondrocytes was downregulated by HT-2 treatment. Moreover, dysregulation of cell cycle was observed; and p53, p21, and CKD6 gene expressions were dysregulated in hiPSC chondrocytes after T-2 toxin treatment. Flow cytometry also demonstrated that there were significantly increased amounts of late apoptotic cells in KBD hiPSC chondrocytes and that the mRNA expression level of Fas was upregulated. In addition, KBD hiPSC chondrocytes presented stronger responses to HT-2 toxin than control hiPSC chondrocytes. These findings confirmed that HT-2 is an environmental risk factor of KBD and that p53 pathway interacted with HT-2 toxin, causing damaged ultrastructure of organelles, accelerating cell cycle in G1 phase, and increasing late apoptosis in KBD hiPSC chondrocytes.

Highlights

Kashin–Beck disease (KBD) is an endemic articular disease and mainly affects people living in southeast Siberia of Russia, the northern region of North Korea, and the long narrow zone from northeastern to southwestern of China (Yu et al, 2019b; Wang et al, 2020)
We used healthy and KBD human induced pluripotent stem cells (hiPSCs) cell lines to try to find out the differences in response to environmental risk factor of KBD, HT-2 toxin
In order to compare the toxicity of HT-2 toxin on differentiating control hiPSC chondrocytes and KBD hiPSC chondrocytes, we first adopted transmission electron microscopy (TEM) to observe the ultrastructural changes of cellular organelles after HT-2 toxin exposure

Summary

Introduction

Kashin–Beck disease (KBD) is an endemic articular disease and mainly affects people living in southeast Siberia of Russia, the northern region of North Korea, and the long narrow zone from northeastern to southwestern of China (Yu et al, 2019b; Wang et al, 2020). KBD mainly damages the epiphyseal growth plate and causes necrosis and apoptosis of chondrocytes in the deep zone of the cartilage (Guo et al, 2014). The damaged epiphyseal growth plate and chondrocytes mostly result in short stature and a secondary chronic osteoarthropathy presenting enlarged and deformed joints (Xiong, 2001). Articular cartilage damage may be achieved by the models, they do not produce multiple foci in the deep zone but extend to the full depth of cartilage (Wang et al, 2011). The etiological research of KBD has been hampered due to lack of animal and cellular disease models

Methods

Results

Conclusion