Analysis And Potential Functional Characterization Of Long Noncoding RNAs Associated With Familial Acne Inversa With NCSTN Mutation

Background:Acne inverse(Acne inversa,AI)is a chronic recurrent inflammatory skin disease that mainly involves apocrine glands.AI can be divided into two types:familial and sporadic and the pathogenesis of familial AI is related to the mutation of γsecretase gene.At present,the most known mutation site is NCSTN subunit of γsecretase.As long noncoding RNA,lncRNA is associated with a variety of inflammatory skin diseases.such as psoriasis,atopic dermatitis and so on.However,little is known about the expression profile and regulatory function of lncRNA in familial AI patients with NCSTN mutation.Therefore,the purpose of this study is to sequence lncRNA in skin tissues between familial AI patients with NCSTN mutation and normal controls by second-generation sequencing technique,and to carry out bioinformatics analysis to explore the regulatory function of lncRNA in familial AI patients,and to find the possible pathogenesis of familial AI.Methods:1.The skin tissues between 3 familial AI patients with NCSTN mutation and 4 normal controls were selected.After the extraction of RNA,the differentially expressed lncRNAs and mRNAs in familial AI patients were identified by second-generation sequencing,and qRT-PCR was performed to validate the 8 lncRNAs with the most significant differential expression.2.The differentially expressed mRNAs targeted by markedly differentially expressed lncRNAs was analyzed by GO and KEGG analysis,to speculate the pathways and regulatory functions that they may be involved in.3.Based on the results of previous studies on the differentially expressed miRNAs between patients with familial AI and normal controls,we integrated the differentially expressed miRNA-mRNA pairs and miRNA-lncRNA pairs from database and constructed a complex lncRNA-miRNA-mRNA regulatory network to study the CeRNA regulation mechanism of lncRNA.4.Constructed the NCSTN keratinocyte-specific knockout mice model and implemented second-generation sequencing.Then,sequence conservation analysis of lncRNAs between patients with familial AI patients with NCSTN mutation and NCSTNΔKC mice was carry out to investigate the orthologues of lncRNAs.Results:1.In total,359 lncRNAs and 1863 mRNAs were differentially expressed between the two groups,The results of qRT-PCR verification showed that the expression trend of the screened lncRNAs was consistent with that observed in the secondgeneration sequencing data.2.GO and KEGG analysis revealed that the dysregulated mRNAs targeted by lncRNAs were mainly associated with the immune regulation,Staphylococcus aureus infection and B cell receptor signalling pathways.3.Previous studies showed that 20 miRNAs were differentially expressed between familial AI patients and normal controls.The lncRNA-miRNA-mRNA coexpression network contained 265 network pairs comprising 55 dysregulated lncRNAs,11 miRNAs and 74 mRNAs.Furthermore,four important lncRNA-miRNA-mRNA CeRNA subnetworks were found.4.Conservation analysis of the differentially expressed lncRNAs between familial AI patients with NCSTN mutation and NCSTNΔKC mice identified 6 lncRNAs with sequence conservation.Conclusion:1.There was a significant difference in lncRNA expression between familial AI patients with NCSTN mutation and normal people.2.Cell phagocytosis,immune regulation and B cell receptor signal pathway may play an important role in familial AI.3.lncRNA regulates gene expression through the formation of gene expression regulatory network with miRNA and mRNA,so as to affect the important link of AI pathogenesis.4.The homologous lncRNAs between familial AI patients and NCSTNΔKC mice may participate in apoptosis,proliferation and skin barrier function.All these findings provide a direction for exploring the regulatory mechanisms underlying the progression of familial AI patients with NCSTN mutation.