Differential Expression Of Inflammatory Genes Research Articles

IL-1β enhances cell viability and decreases 5-FU sensitivity in novel colon cancer cell lines derived from African American patients.

In the U.S., African Americans (AAs) present with the highest incidence and mortality rates for Colorectal Cancer (CRC). When compared to Caucasian American (CA) patients, AAs also have reduced response to the first line standard of care chemotherapeutic agent 5-Fluorouracil (5-FU). Previously, we observed differential gene expression between the two populations, suggesting that colon tumors from AA patients display a decreased antitumor immune response and an increased expression of genes encoding proteins involved in inflammatory processes, such as Interleukin-1β (IL-1β). Here, we investigate the role of IL-1β in modifying chemotherapeutic response and altering expression of proteins in novel AA and well-established CA colon cancer cell lines. RNA sequencing analysis was performed to detect expression of genes involved in inflammation in AA and CA colon cancer cells. The effects of IL-1β on 5-FU response was evaluated by assessing cell viability (MTS assay) and apoptosis (flow cytometry analysis) following treatment with 5-FU alone or in combination with the cytokine. Further, we used an IL-1 receptor antagonist (IL-1Ra) to inhibit IL-1β-induced effects on 5-FU sensitivity and NF-kB pathway activation. AA colon cancer cell lines present significant increase in expression of genes IL1R2 (373-fold change (FC), IRAK1 (3.24 FC), IKBKB, (5.33 FC) NF-KB IA (5.95 FC), MYD88, (3.72 FC), IRAK3 (161 FC), TRAF5 (4.1 FC). A significant decrease in the response to 5-FU treatment, as well as a significant increase in phosphorylation of IκBα and secretion of IL-8, was seen following IL-1β treatment, in both AA and CA cell lines. Finally, treatment with IL-1Ra was able to reverse the effects induced by IL-1β, by increasing the cells sensitivity to 5-FU. IL-1Ra also inhibited phosphorylation of IκBα and IL-8 secretion. Our results suggest a differential expression of inflammatory genes and proteins that might regulate the different response to IL-1β between AA and CA colon cancer cell lines. Our data also demonstrates that IL-1β is involved in modulating 5-FU response in both AA and CA colon cancer cell lines. Further investigation of these mechanisms might help elucidate the differences seen in incidence, mortality and response to therapy in AA colon cancer patients.

Contribution of diabetes mellitus to periodontal inflammation during orthodontic tooth movement.

This study aims to clarify the effects of diabetes mellitus (DM) on inflammatory profile during orthodontic tooth movement (OTM) and explore potential mechanisms. OTM models were established in healthy (Ctrl) and DM rats for 0, 3, 7 or 14 days. The tooth movement distance and bone structural parameters were analyzed through micro-CT. The bone resorption activity and periodontal inflammation status were evaluated through histological staining. RNA sequencing was performed to detect differentially expressed genes in force loading-treated periodontal ligament fibroblasts (PDLFs) with or without high glucose. The differential expression of inflammatory genes associated with NOD-like receptor family pyrin domain containing 3 (NLRP3) between groups was tested in vitro and in vivo. DM caused remarkable reduction of alveolar bone height and density around the moved tooth, corresponding with the higher bone resorption activity and inflammatory scores of DM group. For force loading-treated PDLFs, high glucose induced the activation of inflammatory pathways, including NLRP3. Elevated expression of NLRP3 and cascade molecules (Caspase-1, GSDMD, and IL-1β) were validated by RT-qPCR, Western blot, and immunohistochemistry staining. DM alters the inflammatory status of periodontium and affects tissue reconstruction during OTM. NLRP3 inflammasome may involve in diabetes-induced periodontal changes.

Physical Activity Moderates Inflammatory Gene Expression in Rheumatoid Arthritis

Background: While general population studies have shown associations between greater physical activity and lower inflammatory markers, effects of physical activity on systemic inflammation in immune-mediated inflammatory diseases such as rheumatoid arthritis (RA) are unknown. We examined whether physical activity associates with differential expression of inflammatory genes in RA. Methods: Data derived from an observational RA cohort. Physical activity was measured with the GT9X ActiGraph Link for 7 consecutive 24-hour periods. Clinical data and peripheral whole blood for RNA sequencing were collected. Physical activity groups were defined by the highest and lowest tertiles of moderate/vigorous activity (metabolic equivalent level ≥2.00). RNA extraction, library preparation, Illumina sequencing, and gene expression data processing were performed using established methods. Genes differentially expressed in the most versus least active groups were identified using DESeq2 with the Independent Hypothesis Weighting (IHW) multiple testing procedure and adjusting for sex, age, and race/ethnicity. Ingenuity Pathway Analysis (IPA) Canonical Pathway and Upstream Regulator Analyses were employed on differentially expressed genes with a P<0·1 ranked by log2-fold change to identify biological pathways affected by physical activity. Findings: 35 participants were included (mean age 56±12 years; 91% female; race/ethnicity 31% white, 9% African American, 9% Asian, 40% Hispanic). 767 genes were differentially expressed (padj<0.1) between high versus low activity groups. The high activity group exhibited downregulation of innate and adaptive immune signaling pathways implicated in RA pathogenesis including CD40, STAT3, TREM-1, IL-17a, IL-8, toll-like receptor and interferon signaling. Upstream cytokine activation states demonstrated lower TNF-alpha and interferon among individuals in the high vs. low activity group. Interpretation: RA patients who were more physically active had lower expression of genes involved in innate and adaptive immune signaling compared to those who were more sedentary, providing mechanistic evidence that physical activity may confer protective effects on RA disease outcomes. Funding: This work is supported by NIH/NIAMS R01 AR069616 and NIH/NHLBI K23HL138461-01A1. Additional support from the Rheumatology Research Foundation. Declaration of Interest: None to declare. Ethical Approval: The UCSF Institutional Review Board approved all procedures (UCSF IRB protocol #17-21790) and all participants provided written informed consent.

Differential expression of pro-inflammatory and anti-inflammatory genes of layer chicken bursa after experimental infection with infectious bursal disease virus

Infectious bursal disease (IBD) is one of the most prevalent infectious diseases caused by IBD virus (IBDV), which results in bursal necrosis and immunosuppression that cause severe damage to the immune system in chickens. Cytokines are important mediators and regulators of both types of host responses. In the present study, layer chickens were artificially challenged with IBDV, and the differential expression of inflammatory genes was explored by using quantitative real-time PCR, which offered basic data for further study of IBDV pathogenesis. Data showed that after IBDV infection, the virus load in the bursa of Fabricius (BF) peaked at 96h and then gradually decreased. Compared with those of the negative-infected group, the mRNA expression levels of pro-inflammatory cytokines (interleukin [IL]-1β, IL-6, IL-7, IL-8, tumor necrosis factor [TNF]-α, transforming growth factor [TGF]-β) and anti-inflammatory cytokine IL-10 in the infected group increased to varying degrees at 12 to 192h, respectively. Furthermore, the IL-1β mRNA expression peaked at 48h; the mRNA transcript levels of IL-6, IL-8, and IL-10 were the highest at 96h; TNF-α mRNA expression peaked at 120h; the IL-7 mRNA expression peaked at 144h; and the TGF-β mRNA transcript level was the highest at 192h. Taken together, these observations indicated that along with the change pattern of IBDV proliferation in BF, the mRNA expression of cytokines (IL-1β, IL-6, IL-7, IL-8, IL-10, TNF-α, TGF-β) obviously increased, and the kinetics of each of these cytokines was different. The kinetics of IL-6/IL-10 mRNA expression ratio was significantly positively correlated with that of the virus load. These results suggest that IBDV infection seriously interferes with the natural immune response mediated by inflammatory cytokines in chickens.

Differential transcriptome expression in human nucleus accumbens as a function of loneliness.

Loneliness is associated with impaired mental and physical health. Studies of lonely individuals reported differential expression of inflammatory genes in peripheral leukocytes and diminished activation in brain reward regions such as nucleus accumbens, but could not address gene expression in the human brain. Here, we examined genome-wide RNA expression in postmortem nucleus accumbens from donors (N = 26) with known loneliness measures. Loneliness was associated with 1 710 differentially expressed transcripts from 1 599 genes (DEGs; FDR p < 0.05, fold-change ≥ |2|, controlling for confounds) previously associated with behavioral processes, neurological disease, psychological disorders, cancer, organismal injury, and skeletal and muscular disorders, as well as networks of upstream RNA regulators. Furthermore, a number of DEGs were associated with Alzheimer’s disease genes (which was correlated with loneliness in this sample, although gene expression analyses controlled for AD diagnosis). These results identify novel targets for future mechanistic studies of gene networks in nucleus accumbens and gene regulatory mechanisms across a variety of diseases exacerbated by loneliness.

Diet high in retinoic acid controls M1/M2 activation phenotypes in macrophages and protects from monocrotaline-induced pulmonary fibrosis

Monocrotaline (MCT) causes endothelial damage and monocyte/macrophage infiltration in the lungs, effects that further aggravate the destructive process via excessive release of potent inflammatory mediators. We used cDNA GEArray to analyze differential expression of inflammatory genes in alveolar macrophages, which may be attributed to MCT-induced disease. Most of the genes regulated in response to MCT were consistent with the M2 phenotype of alternatively activated macrophages, suggesting their role in pulmonary fibrosis. A diet high in retinoic acid (RA) prevented MCT-induced monocyte/macrophage infiltration and severe lung injury. At the transcriptional level, RA reprogrammed the inflammatory expression patterns induced in macrophages by MCT. The identified sets of RA-regulated genes associate alveolar macrophages with classical M1 activation phenotype. Collectively, our GEArray data provide new insights into regulatory role of RA nutritional status in the transcriptional modulation of inflammatory responses in alveolar macrophages, and improve our understanding of cytokine/chemokine imbalance in the pathogenesis of pulmonary fibrosis.