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Translational regulation contributes to the secretory response of chondrocytic cells following exposure to Interleukin-1B

ABSTRACT: Chondrocytes undergo changes to their protein translational capacity during osteoarthritis progression, but a study of how disease-relevant signals affect chondrocyte protein translation at the transcriptome level has not previously been performed. In this study we describe how the inflammatory cytokine IL-1B rapidly affects protein translation in the chondrosarcoma cell line SW1353. Using ribosome profiling we demonstrate that IL-1B induced altered translation of inflammatory-associated transcripts through differential translation and the use of multiple open reading frames. Proteomic analysis of the cellular layer and the conditioned media of these cells identified that proteins which were differentially translated were most readily detected in the secretome. We have produced combined ribosome profiling and proteomic datasets which provide a valuable resource in understanding the processes that are occuring during cytokine stimulation of chondrocytic cells.

OTHER RELATED OMICS DATASETS IN: MSV000084104

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Fibroblast

DISEASE(S): Disease Free

SUBMITTER: Ben McDermott

LAB HEAD: Dr Simon Tew

PROVIDER: PXD012985 | Pride | 2019-07-11

REPOSITORIES: Pride

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Translational regulation contributes to the secretory response of chondrocytic cells following exposure to interleukin-1β.

McDermott Benjamin T BT Peffers Mandy J MJ McDonagh Brian B Tew Simon R SR

The Journal of biological chemistry 20190712 35

Osteoarthritis is a chronic disease characterized by the loss of articular cartilage in synovial joints through a process of extracellular matrix destruction that is strongly associated with inflammatory stimuli. Chondrocytes undergo changes to their protein translational capacity during osteoarthritis, but a study of how disease-relevant signals affect chondrocyte protein translation at the transcriptomic level has not previously been performed. In this study, we describe how the inflammatory c ...[more]

PMID: 31300557

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Project description:IL-1B is an important cytokine that is often found to be up-regulated during osteoarthritic and rheumatoid joint diseases. It is viewed as a catabolic factor, inducing enzymes that allow for the degradation of the cartilage extracellular matrix and also has essential roles as an autocrine and paracrine factor in fibronectin fragment-mediated degradation. It can also reduce the synthesis of the major cartilage components, type II collagen and aggrecan. On the other hand, IL-1B also has the ability to induce the growth and morphogenic factor BMP-2. During joint diseases, IL-1B is synthesized by both synovial cells and chondrocytes. Addition of IL-1 biological antagonists such as IL-1 receptor antagonists can suppress cartilage degradation in vitro. Thus, the production of IL-1B could act as the first step in mediating a cascade of other mediators in cartilage which could be relevant to the fate of the cartilage. In order to obtain a global picture of the effect of IL-1B production on human adult articular chondrocytes, we analyzed changes in gene expression induced by IL-1B by microarray analysis. We found that IL-1B has a diverse effect on gene expression profile in chondrocytes. One of the predominant responses that we observed in adult human articular chondrocytes on exposure to IL-1B is a dramatic increase in a large set of chemokines and other genes related to the inflammatory cascade. Keywords: Gene response to IL-1B (10 ng/ml) Cartilage was obtained from adult human tissue donors with above the knee amputations due to chondrosarcoma or traumatic injury or from autopsy. Chondrocytes were isolated following established protocols, maintained in high density, and treated with IL-1B (10 ng/ml). Chondrocytes treated with buffer only served as the untreated control. The experiment was carried out in duplicate. Total RNA was extracted from these chondrocytes, labeled with fluorophores (Cy3 or Cy5) and analyzed for expression changes using the Human Operon/Qiagen v3.0 oligonucleotide array. The analysis was repeated with the fluorophore dyes exchanged between the untreated and experimental RNAs.

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