The Study Of Associated Genes And Molecular Biological Characteristics In SLE With Multisystem Damage Pathogenisis

Part IA pilot study of gene testing of osteonecrosis of the femoral head in systemic lupus erythematosus using targeted next-generation sequencingBackgroundPrevious publications indicated that genetic predisposition might play important roles in the onset of osteonecrosis of the femoral head(ONFH)in systemic lupus erythematosus(SLE).For example,some gene loci such as CR2(complement C3d receptor 2),NOS3(nitric oxide synthase 3),COL2A1(collagen type ? alpha 1 chain),PTPN22(protein tyrosine phosphatase non-receptor type 22)and TRPV4(transient receptor potential cation channel subfamily V member 4)were reported to be involved in this process.Therefore,we investigated whether the risk of ONFH in SLE is associated with single nucleotide variations(SNVs)in these five genes.MethodsSNVs in the CR2,NOS3,COL2A1,PTPN22 and TRPV4 genes were examined by using FastTarget and Illumina Miseq sequencing technologies in 49 cases of SLE with ONFH.BWA was used to align the sequencing reads to hgl9 and GATK and Varscan programs were used to perform SNV calling.PolyPhen-2,SIFT,and MutationTaster assessed the functional effects of non-synonymous SNVs.ResultsSix of the 49 patients were confirmed to have low frequency SNVs,including one patient with SNVs in NOS3(exon6:c.814G>A:p.E272K and exon7:c.814G>A:p.E272K.),four in COL2A1(rs41263847:exon29:c.1913C>T:p.T638I,exon28:c.1706C>T:p.T569I,and rs371445823:exon8:c.580G>A:p.A194T,exon7:c.373G>A:p.A125T)and one in CR2(rs45573035:exon2:c.200C>G:p.T67S).ConclusionsThe onset of ONFH in SLE might be associated with the identified SNVs in NOS3,COL2A1,and CR2.These findings may have important pharmacogenetic implications.However,the detailed mechanisms of the associations need to be determined in further studies.Part ?Collagen triple helix repeat containing-1:a novel biomarker associated with disease activity in Systemic lupus erythematosusBackgroundCollagen triple helix repeat containing-1(CTHRC1)is highly upregulated in solid tumors,and many studies uncovered the effect of CTHRC1 on bone mass,myelination,synoviocytes and collagen synthesis in keloid fibroblasts.Additionally,CTHRC1 may be involved in the progres-sion of rheumatoid arthritis(RA).The objective of this article is to investigate whether the aberrant expression of collagen triple helix repeat containing-1(CTHRC1)from patients with systemic lupus erythematosus(SLE)could contribute to the pathogenesis of lupus.MethodsWe divided SLE patients into active groups(Systemic Lupus Erythematosus Disease Activity Index(SLEDAI)? 6)and inactive groups(SLEDAI score<6).Serum concentrations of CTHRC1,interferon alpha,interleukin(IL)-28A and IL-28B were determined using an enzyme-linked immunosorbent assay in a group of 40 patients with SLE.Results were compared with those from 23 healthy controls.ResultsSerum CTHRC1 protein levels were higher in patients with SLE compared with healthy controls.Patients with active disease displayed higher CTHRC1 levels compared with those with inactive disease as well.There was a positive association between serum CTHRC1 levels and SLEDAI and erythrocyte sedimentation rate,and a negative correlation with complement 3 and 4.Moreover,serum CTHRC1 levels were higher in SLE patients with arthritis and anemia compared with patients without the above-mentioned manifestations.ConclusionsThese findings indicate CTHRC1 probably plays an important part in the pathogenesis of SLE,and is positively associated with disease activity,while it also likely refers to the development of arthritis and anemia in SLE.Therefore,CTHRC1 may provide a novel research target and shed new light on the pathogenesis and therapy of SLE.Part ?MicroRNA-223 down regulate the NLRP3 expression of PBMC in the pathogenesis of SLEBackgroundThe innate immune system and the adaptive immune system play an important role in the pathogenesis of(Systemic lupus erythematosus,SLE)?Studies have found that MicroRNA-223(miR-223)have important effect in the development of immune system and immune related disease?So far there is little research about miR-223 take part in the pathogenesis of SLE,and no about it regulate the NLRP3 in the pathogenesis of SLE.To determinate the mRNA levels of MicroRNA-223 and NLRP3 inflammasome components(NLRP3,ASC and Caspase-1)in PBMC from patients with SLE and to explore its clinical significance.MethodsReal time PCR was used to detect the mRNA levels of miR-223 and NLRP3 inflammasome components in PBMC from 35 SLE patients and 30 healthy controls and evaluate its correlations with clinical and laboratory parameters(anti-dsDNA,anti-AnuA,ESR and SLEDAI).ResultsCompared with healthy controls,mRNA levels of miR-223 and Caspase-1 were higher in the PBMC from SLE patients,but the NLRP3 and ASC mRNA expression levels were in the opposite trend.Correlation analysis results showed that there was a negative correlation of mRNA levels between the miR-223 and NLRP3,but the miR-223 had no correlation with ASC and Caspase-1.The mRNA levels of miR-223 were positively correlated with anti-dsDNA,SLEDAI and ESR,but had no correlation with anti-AnuA.There was no significant difference in the expression of miR-223 in PBMC between LN patients and SLE patients without renal impairment.ConclusionIn PBMC from SLE patients,miR-223 and mRNA levels of Caspase-1 were increased,while mRNA levels of NLRP3 and ASC were decreased,and the level of miR-223 was related to the disease activity.Negative regulation of NLRP3 by miR-223 plays an important role in the pathogenesis of SLE.