Global Gene Expression Profiling and Alternative Splicing Events during the Chondrogenic Differentiation of Human Cartilage Endplate-Derived Stem Cells.

Jin Shang,Huan Liu,Lei Shangguan,Xin Fan,Yue Zhou

doi:10.1155/2015/604972

Abstract

Low back pain (LBP) is a very prevalent disease and degenerative disc diseases (DDDs) usually account for the LBP. However, the pathogenesis of DDDs is complicated and difficult to elucidate. Alternative splicing is a sophisticated regulatory process which greatly increases cellular complexity and phenotypic diversity of eukaryotic organisms. In addition, the cartilage endplate-derived stem cells have been discovered and identified by our research group. In this paper, we continue to investigate gene expression profiling and alternative splicing events during chondrogenic differentiation of cartilage endplate-derived stem cells. We adopted Affymetrix Human Transcriptome Array 2.0 (HTA 2.0) to compare the transcriptional and splicing changes between the control and differentiated samples. RT-PCR and quantitative PCR are used to validate the microarray results. The GO and KEGG pathway analysis was also performed. After bioinformatics analysis of the data, we detected 1953 differentially expressed genes. In terms of alternative splicing, the Splicing Index algorithm was used to select alternatively spliced genes. We detected 4411 alternatively spliced genes. GO and KEGG pathway analysis also revealed several functionally involved biological processes and signaling pathways. To our knowledge, this is the first study to investigate the alternative splicing mechanisms in chondrogenic differentiation of stem cells on a genome-wide scale.

Highlights

Low back pain (LBP) is one of the most prevalent diseases which needs medical advice and results in chronic disabilities [1]
Since the necessary nutrients have to diffuse across the intervertebral cartilage endplate (CEP) to supply the intervertebral discs (IVDs), many researchers speculate that CEP degeneration play critical roles in the initiation and development of degenerative disc diseases (DDDs) [8, 9]
The proteoglycan loss from CEP is closely related to proteoglycan loss in nucleus pulposus (NP), in turn resulting in DDD [13]

Summary

Introduction

Low back pain (LBP) is one of the most prevalent diseases which needs medical advice and results in chronic disabilities [1]. Degenerative disc disease (DDD) is a common reason for LBP [3]. Since the necessary nutrients have to diffuse across the intervertebral cartilage endplate (CEP) to supply the intervertebral discs (IVDs), many researchers speculate that CEP degeneration play critical roles in the initiation and development of DDD [8, 9]. The CEP degeneration has several manifestations, such as proteoglycan loss [10], CEP calcification [11], and ECM synthesis defects [12]. Calcification or sclerosis of CEP reduced the diffusion ability of nutrient molecules into adjacent disc, leading to DDD [14]. It is crucial to illuminate the mechanisms of CEP degeneration and DDD for developing effective therapies

Methods

Results

Conclusion