Transcriptome analysis reveals specific changes in osteoarthritis synovial fibroblasts

Rey, Manuel; Usategui, Alicia; Izquierdo, Elena; Cañete, Juan D.; Blanco, Francisco J.; Criado, Gabriel; Pablos, José L.

doi:10.1136/annrheumdis-2011-200281

Cited by 39 publications

(27 citation statements)

References 41 publications

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“…Subsequent functional studies showed that LBH expression in FLS is regulated by platelet‐derived growth factor, a major growth factor that contributes to synovial lining hyperplasia in RA . Low levels of LBH expression, which have previously been documented for RA FLS , regulates the cell cycle and can contribute to a hyperproliferative state of RA FLS. Understanding why low LBH expression levels could participate in RA pathogenesis can provide insight into disease mechanisms and possibly novel therapeutic targets.…”

Section: Discussionmentioning

confidence: 93%

LBH Gene Transcription Regulation by the Interplay of an Enhancer Risk Allele and DNA Methylation in Rheumatoid Arthritis

Hammaker

Whitaker

Maeshima

et al. 2016

Arthritis & Rheumatology

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Objective To identify non-obvious therapeutic targets for rheumatoid arthritis, we performed an integrative analysis incorporating multiple ‘omics data and the ENCODE database for potential regulatory regions. This analysis identified the Limb Bud and Heart development (LBH) gene, which has risk alleles associated with RA/celiac disease and lupus, and can regulate cell proliferation in RA. We identified a novel LBH transcriptional enhancer with an RA-risk allele (rs906868 G (risk, Ref) /T) 6kb upstream of the LBH gene with a differentially methylated locus. The confluence of three regulatory elements, rs906868, an RA differentially methylated locus and a putative enhancer, led us to investigate their effect on LBH regulation in fibroblast-like synoviocytes (FLS). Methods We cloned the 1.4kb putative enhancer with either the rs906868 Ref allele or SNP variant into reporter constructs. The constructs were methylated in vitro and transfected into cultured FLS by nucleofection. Results We found that both variants increased transcription, thereby confirming the region’s enhancer function. Unexpectedly, the transcriptional activity of the Ref risk allele was significantly lower than the SNP variant and is consistent with low LBH levels as a risk factor for aggressive FLS behavior. Using RA FLS lines with homozygous Ref or SNP allele, we confirmed that homozygous Ref lines expressed lower LBH mRNA levels than the SNP lines. Methylation significantly reduced enhancer activity for both alleles, indicating that enhancer function is dependent on its methylation status. Conclusion This study shows how the interplay between genetics and epigenetics can affect expression of LBH in rheumatoid arthritis.

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Section: Discussionmentioning

confidence: 93%

LBH Gene Transcription Regulation by the Interplay of an Enhancer Risk Allele and DNA Methylation in Rheumatoid Arthritis

Hammaker

Whitaker

Maeshima

et al. 2016

Arthritis & Rheumatology

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“…For each analysis, eight raw fluorescence data files with gpr extension, obtained after scanning and corresponding to four exposed cultures and four control cultures, were submitted to GeneSpring software GX11 (Agilent Technologies) as follows. Concerning the statistics methodology, we used a widespread method for determining the significance change of gene expression [32]. The raw data were first normalized using the percentile shift 75 normalization method.…”

Section: Methodsmentioning

confidence: 99%

Intestinal toxicity evaluation of TiO2 degraded surface-treated nanoparticles: a combined physico-chemical and toxicogenomics approach in caco-2 cells

et al. 2012

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BackgroundTitanium dioxide (TiO2) nanoparticles (NPs) are widely used due to their specific properties, like UV filters in sunscreen. In that particular case TiO2 NPs are surface modified to avoid photocatalytic effects. These surface-treated nanoparticles (STNPs) spread in the environment and might release NPs as degradation residues. Indeed, degradation by the environment (exposure to UV, water and air contact …) will occur and could profoundly alter the physicochemical properties of STNPs such as chemistry, size, shape, surface structure and dispersion that are important parameters for toxicity. Although the toxicity of surface unmodified TiO2 NPs has been documented, nothing was done about degraded TiO2 STNPs which are the most likely to be encountered in environment. The superoxide production by aged STNPs suspensions was tested and compared to surface unmodified TiO2 NPs. We investigated the possible toxicity of commercialized STNPs, degraded by environmental conditions, on human intestinal epithelial cells. STNPs sizes and shape were characterized and viability tests were performed on Caco-2 cells exposed to STNPs. The exposed cells were imaged with SEM and STNPs internalization was researched by TEM. Gene expression microarray analyses were performed to look for potential changes in cellular functions.ResultsThe production of reactive oxygen species was detected with surface unmodified TiO2 NPs but not with STNPs or their residues. Through three different toxicity assays, the STNPs tested, which have a strong tendency to aggregate in complex media, showed no toxic effect in Caco-2 cells after exposures to STNPs up to 100 μg/mL over 4 h, 24 h and 72 h. The cell morphology remained intact, attested by SEM, and internalization of STNPs was not seen by TEM. Moreover gene expression analysis using pangenomic oligomicroarrays (4x 44000 genes) did not show any change versus unexposed cells after exposure to 10 μg/ mL, which is much higher than potential environmental concentrations.ConclusionsTiO2 STNPs, degraded or not, are not harmful to Caco-2 cells and are unlikely to penetrate the body via oral route. It is likely that the strong persistence of the aluminium hydroxide layer surrounding these nanoparticles protects the cells from a direct contact with the potentially phototoxic TiO2 core.

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“…Many investigators have explored gene expression changes in OA by transcriptomic profiling of human joint tissues (). While these studies have yielded important information on candidate genes in advanced OA, there are drawbacks to using patient tissue for expression profiling.…”

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confidence: 99%

Transcriptomics of Wild‐Type Mice and Mice Lacking ADAMTS‐5 Activity Identifies Genes Involved in Osteoarthritis Initiation and Cartilage Destruction

Bateman

Rowley

Belluoccio

et al. 2013

Arthritis & Rheumatism

110

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Objective. To identify changes in gene expression in mice with osteoarthritis (OA) in order to explore the mechanisms of the disease.Methods. Gene expression profiling was performed in cartilage from mice with surgically induced OA. We used wild-type (WT) mice and Adamts5⌬cat mice, in which ADAMTS-5 activity is lacking and aggrecan loss and cartilage erosion are inhibited, to distinguish gene expression changes that are independent of ADAMTS-5 activity and cartilage breakdown. Mechanical instability was introduced into the knee joints of 10-week-old male mice via surgical destabilization of the medial meniscus (DMM). Cartilage from the developing lesion in the destabilized medial meniscus and corresponding regions in sham-operated joints was harvested by microdissection at 1, 2, and 6 weeks postsurgery, and RNA was extracted, amplified, and hybridized to wholegenome microarrays.Results. Several previously identified OA-related genes, including Ptgs2, Crlf1, and Inhba, and novel genes, such as Phdla2 and Il11, were up-regulated in both WT mice and Adamts5⌬cat mice, indicating that they are independent of ADAMTS-5 activity. The altered expression of other genes, including Col10a1, the sentinel marker of cartilage hypertrophy, and Wnt/␤-catenin pathway genes, required ADAMTS-5 activity. Cell death pathway genes were dysregulated, and Tp53, Foxo4, and Xbp1 endoplasmic reticulum-stress transcriptional networks were activated. Analysis of degradome genes identified up-regulation of many proteases, including Mmp3, Capn2, and the novel cartilage proteases Prss46 and Klk8. Comparison with other studies identified 16 genes also dysregulated in rat and human OA as priorities for study.Conclusion. We have identified, for the first time, several genes that have an ADAMTS-5-independent role in OA, identifying them as possible OA initiation candidates. This work provides new insights into the sequence of gene dysregulation and the molecular basis of cartilage destruction in OA.

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Transcriptome analysis reveals specific changes in osteoarthritis synovial fibroblasts

Cited by 39 publications

References 41 publications

LBH Gene Transcription Regulation by the Interplay of an Enhancer Risk Allele and DNA Methylation in Rheumatoid Arthritis

LBH Gene Transcription Regulation by the Interplay of an Enhancer Risk Allele and DNA Methylation in Rheumatoid Arthritis

Intestinal toxicity evaluation of TiO2 degraded surface-treated nanoparticles: a combined physico-chemical and toxicogenomics approach in caco-2 cells

Transcriptomics of Wild‐Type Mice and Mice Lacking ADAMTS‐5 Activity Identifies Genes Involved in Osteoarthritis Initiation and Cartilage Destruction

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