Micro RNA Analysis of the Ossification of Posterior Longitudinal Ligament (OPLL).
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ABSTRACT: Proteomics analysis was conducted using sera from 31 OPLL patients and 17 healthy individuals, detecting a significantly defective protein in the former as compared with the latter. The continuous and mixed types of OPLL were compared, demonstrating a lower level of the protein in the former than the latter. Clinically, the continuous type, i.e., the ossification of the entire longitudinal ligament, was frequently detected in severe cases.
Project description:Ossification of the posterior longitudinal ligament(OPLL) is a progressive abnormal calcification of the spinal ligament, which causes myelopathy and neurological symptoms. Both genetic and environmental factors play an important role in its occurrence and progress. In recent decades, many studies on OPLL have shown that it is a multifactorial disease. However, the role of circRNAs in the pathogenesis of OPLL is far from clear. In order to identify the transcriptional regulators of OPLL, we compared the expression of circRNAs in the posterior longitudinal ligament tissues from OPLL patients and healthy volunteers through microarray analysis. The analysis revealed a set of circRNAs specifically regulated in humans with heterotopic ossification of ligament tissue. These findings imply that circRNAs may play an important role in OPLL, which provides new ideas for the study of OPLL.
Project description:The initiation and/or progression of ossification of the posterior longitudinal ligament (OPLL) is associated with cyclic tensile strain, but the pathomechanism of OPLL remains unclear. Indian hedgehog (Ihh) and its related signaling are key factors in normal enchondral ossification. However, the relation of OPLL to Ihh is unclear. The purpose of this study is to investigate the contribution of mechanical strain to OPLL and the relation of Ihh to OPLL. Cultured posterior longitudinal ligament cells were subjected to 24 hours of cyclic tensile strain and then analyzed by microarray.
Project description:The initiation and/or progression of ossification of the posterior longitudinal ligament (OPLL) is associated with cyclic tensile strain, but the pathomechanism of OPLL remains unclear. Indian hedgehog (Ihh) and its related signaling are key factors in normal enchondral ossification. However, the relation of OPLL to Ihh is unclear. The purpose of this study is to investigate the contribution of mechanical strain to OPLL and the relation of Ihh to OPLL.
Project description:We found disease-specific proteins from serum proteomics of ossification of posterior longitudinal ligament, and made knockout mice. We discovered protein peculiar to a disease in serum of the ossification of posterior longitudinal ligament (OPLL), and, as a result of producing knockout mouse, we checked spinal ligament ossification and combination of diabetes and thought that it was a typical mouse and analyzed the sequence of the kidney organization.
Project description:Ossification of the posterior longitudinal ligament (OPLL) is formed by heterogeneous ossification of posterior longitudinal ligament. The patho-mechanism of OPLL is still largely unknown. For the rarity of the primary cell samples, here we immortalized the OPLL patient's posterior longitudinal ligament primary cells and normal patient's posterior longitudinal ligament primary cells. The transcriptome of the cells were compared and we found that the osteogenic ability of the OPLL ligament cells were inherited in the immortalized cells.
Project description:Ectopic bone formation is the chief characteristic of ossification of the posterior longitudinal ligament (OPLL). Emerging evidence has revealed that long non-coding RNAs (lncRNAs) can regulate the osteogenic differentiation of mesenchymal stem cells (MSCs), which are the main cells responsible for bone formation. However, the role of lncRNAs in the pathogenesis of OPLL remains unclear. In this study, 725 aberrantly expressed lncRNAs and 664 mRNAs in osteogenically differentiated MSCs from OPLL patients (OPLL MSCs) were identified by microarrays and confirmed by qRT-PCR assays. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed that the most enriched pathways included the p53, JAK-STAT and PI3K-Akt signaling pathways. The coexpression network showed the interactions between the aberrantly expressed lncRNAs and mRNAs in OPLL MSCs, and the potential targets and transcription factors of the lncRNAs were predicted. Our research demonstrated the aberrantly expressed lncRNA and mRNA and the potential regulatory networks involved in the ectopic bone formation of OPLL. These findings imply that lncRNAs may play a vital role in OPLL, which provides a new perspective on the pathogenesis of OPLL.
Project description:Ossification of the posterior longitudinal ligament (OPLL) is formed by heterogeneous ossification of posterior longitudinal ligament. The patho-mechanism of OPLL is still largely unknown. Recently, disorders of metabolism are thought to be the center of many diseases such as OPLL. Advanced glycation end product (AGE) are accumulated in many extracellular matrixes such as ligament fibers, and it can functions as cellular signal through its receptor (RAGE), contributing to various events such as atherosclerosis or oxidative stress. However, its role in OPLL formation is not yet known. Therefore, we performed high-through-put RNA sequencing on primary posterior longitudinal ligament cells treated with different doses of AGEs (1µM, 5µM and negative control), with or without BMP2 (1µM). mRNA profiles of Primary human posterior longitudinal ligament cells stimulated with various stimuli (Control, 1µM AGE-BSA, 5µM AGE-BSA, 1µM AGE-BSA with BMP2, 5µM AGE-BSA with BMP2) were generated by deep sequencing on Ion Proton
Project description:Ossification of the posterior longitudinal ligament (OPLL) is formed by heterogeneous ossification of posterior longitudinal ligament. The patho-mechanism of OPLL is still largely unknown. Recently, disorders of metabolism are thought to be the center of many diseases such as OPLL. Advanced glycation end product (AGE) are accumulated in many extracellular matrixes such as ligament fibers, and it can functions as cellular signal through its receptor (RAGE), contributing to various events such as atherosclerosis or oxidative stress. However, its role in OPLL formation is not yet known. Therefore, we performed high-through-put RNA sequencing on primary posterior longitudinal ligament cells treated with different doses of AGEs (1µM, 5µM and negative control), with or without BMP2 (1µM).
Project description:Ossification of the posterior longitudinal ligament (OPLL) of the spine is characterized by progressive ectopic bone formation in the spinal ligament. To identify the genes related to ectopic ossification of human spinal ligament affected by mechanical stress, analyses using cDNA microarray were carried out using cultured human spinal ligament cells that had been subjected to uniaxial cyclic stretching. cDNA microarrays revealed that ranges of distribution of both up- and down-regulated genes evoked by cyclic stretching were significantly wider in cells from than in non-OPLL cells. Keywords: mechanical-stress response
Project description:Ossification of the posterior longitudinal ligament (OPLL) is formed by heterogeneous ossification of posterior longitudinal ligament. The patho-mechanism of OPLL is still largely unknown. MicroRNAs are small nucleatides that function as regulators of gene expression in almost any biological process. However, few microRNAs are reported to have a role in the pathological process of OPLL. Therefore, we performed high-throughput microRNA sequencing and transcriptome sequencing of primary OPLL and PLL cells in order to decipher the interacting network of microRNAs in OPLL. MRNA and microRNA profiles were done using primary culture cells of human ossification of the posterior longitudinal ligament (OPLL) tissue and normal posterior longitudinal ligament (PLL) tissue.