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.
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:<p>Background: Chronic obstructive pulmonary disease (COPD) is a progressive inflammatory disorder with rising global morbidity and mortality. Emerging evidence suggests that systemic metabolic alterations, particularly dyslipidemia, contribute to COPD pathogenesis. However, the mechanisms linking lipid dysregulation to pulmonary inflammation and tissue injury remain poorly defined.</p><p>Methods: Untargeted metabolomic profiling was performed on plasma samples from healthy individuals and patients with COPD to identify disease associated metabolic alterations. A high-cholesterol diet (HCD) mouse model, with or without chronic cigarette smoke exposure, was used to examine the impact of systemic cholesterol elevation on lung structure and inflammation. Complementary in vitro studies using THP-1 derived and bone marrow derived macrophages were conducted to investigate mitochondrial function, ROS generation, and downstream signaling pathways. Transcriptomic analyses were employed to identify key molecular mediators.</p><p>Results: Plasma metabolomics revealed significant dysregulation of lipid metabolism in COPD, with elevated cholesterol levels correlating inversely with lung function. In vivo, HCD feeding induced pulmonary inflammation and further exacerbated cigarette smoke induced lung tissue destruction. In macrophages, combined cholesterol loading and cigarette smoke extraction treatment disrupted mitochondrial integrity, reduced respiratory capacity, and increase ROS generation. Elevated ROS levels upregulated PPIA, which in turn activated NF-κB signaling and promoted IL-1β secretion. Silencing PPIA or inhibiting ROS significantly attenuated NF-κB activation and cytokine release. Consistent with these findings, increased PPIA expression and NF-κB phosphorylation were observed in lungs of HCD-fed, cigarette smoke exposed mice, and PPIA levels were elevated in bronchoalveolar lavage fluid from COPD patients.</p>
Project description:We analyzed gene expression profiling of lung tissue to define molecular pathway of COPD using recent RNA sequencing technology.Lung tissue was obtained from 98 COPD subjects and 91 subjects with normal spirometry. RNA isolated from these samples was processed with RNA-seq using HiSeq 2000. Gene expression measurements were calculated using Cufflinks software. Differentially expressed genes and isoforms were chosen using t-test. Some of differentially expressed genes were validated by quantitative real-time PCR. Examination of lung tissue in COPD patients versus normal control
Project description:Chronic inflammatory and immune dysregulation are critical drivers in the development and progression of chronic obstructive pulmonary disease (COPD). Posttranslational modifications, such as glycosylation of Immunoglobulin G (IgG), modulates systemic inflammatory homeostasis. This study aims to profile plasma IgG glycopeptides (IgGPs) in COPD patients to uncover new insights into its pathogenesis and to identify novel biomarkers. Plasma IgG N-glycopeptides from 90 COPD patients, 45 clinical defined early COPD (CECOPD) patients and 90 healthy individuals were analyzed using an integrated platform that combines Fe3O4@PDA@DETA nanospheres enrichment with high-resolution mass spectrometry measurement. Correlations between IgG N-glycoforms and clinical parameters were assessed to explore underlying mechanisms of COPD progression. Disease-specific IgGPs were identified in both ECOPD and COPD cohorts. Notably, IgG glyco-pattern, rather than IgG levels, changed with disease progression. Early COPD patients showed decreased bisection and increased site-specific afucosylated galactosylation and fucosylation of IgG, indicating an anti-inflammatory state. In contrast, COPD patients gave increased inflammation, characterized by reduced galactosylation and sialylation. Interestingly, a subset of healthy controls displayed IgGPs patterns similar to early COPD, possibly reflecting the impact of substantial smoking exposure and associated immune responses. These findings suggest that plasma IgG glycosylation could serve as a potential biomarker for early COPD diagnosis, providing valuable insights into immune system changes during disease progression.