Font Size: a A A

Research On The Potential Pathogenic Genes Of Dermatosis Papulosa Nigra Based On Whole Exome Sequencing

Posted on:2023-10-04Degree:MasterType:Thesis
Country:ChinaCandidate:X L WangFull Text:PDF
GTID:2544306614482064Subject:Dermatology and venereology
Abstract/Summary:PDF Full Text Request
BackgroundDermatosis Papulosa Nigra(DPN)is a common benign skin disease on the Fitzpatrick scale IV–VI.The typical clinical lesions are multiple black or brownish-black rounded on the face.Flat papules with no fusion between the skin mounds,about 1 to 5 mm in diameter,and smooth surface[1,2,3].It is easy to be misdiagnosed clinically,and it is often misdiagnosed as seborrheic keratosis,syringoma,flat wart,glossy lichen and other diseases[3,4].The pathogenesis of the disease is currently unclear.The clinical treatment methods are relatively simple,limited to invasive methods such as liquid nitrogen,freezing,carbon dioxide laser and surgery,and there are also obvious side effects such as pigmentation and scarring[4,5].With the gradual improvement of people’s pursuit of a better life and facial aesthetics,the aesthetic disfigurement caused by the disease often causes huge psychological burdens and family pressures on patients.Understanding its occurrence and development mechanism will help in the management and medical practice throughout the process.In recent years,whole exome sequencing(WES)has been widely used in clinical practice,providing an important reference for clinicians in the diagnosis and treatment of tumor and non-tumor diseases[6,7,8].Whole exon sequencing contains about 20,000 genes.The exon regions in the genome are enriched by liquid-phase hybridization capture technology or multiplex PCR amplification technology,and then high-throughput sequencing technology(next generation sequencing)is used to enrich the exon regions in the genome.,NGS)for gene locus detection on the enriched sequences.It can cover more than 98%of point mutations and the vast majority of copy number variants in a very wide range,providing a new detection method for finding disease-related pathogenic mutation genes[8,9].At present,the pathogenic genes related to the pathogenesis of DPN have not been identified,and further related research is needed to fully understand the molecular mechanism of DPN pathogenesis.ObjectiveThe clinical data of DPN cases were collected,and the characteristics of DPN clinical skin lesions,dermoscopy and pathological sections of skin lesions were described in detail.Improve the awareness of DPN disease and reduce the misdiagnosis rate of DPN in clinical practice.Through WES and bioinformatics analysis,we can deeply understand the pathogenesis of the disease at the molecular gene level,search for specific potential DPN pathogenic genes,and find potentially effective therapeutic targets for the follow-up to fundamentally inhibit the occurrence of the disease.Molecular theory basis.In addition,the discovery of germ cell mutations and somatic mutations that affect the structural and functional variation of the protein also provides a molecular biological genetic theoretical basis for the follow-up study of the pathogenesis and genetic mechanism of DPN.MethodsThe clinic collects relevant clinical information of DPN patients,skin lesions manifestations,dermoscopic manifestations,and skin pathological characteristics.And make records,statistical summary of relevant results.Extract 10 DPN patients with clinical manifestations and histopathological diagnosis,extract peripheral blood and extract DNA,at the same time extract the DNA of patients’skin lesions,perform WES on-machine sequencing,and compare the reference genome through quality control of the sequencing data,detection of mutation sites,identification of somatic mutations and germ cell mutations,annotation of mutation sites,comparison with disease gene mutation-related databases,screening of new somatic mutation sites in the whole exon region of DPN;and further screening for potential potential Disease-causing gene mutation sites.For the potential pathogenic gene mutation site,use bioinformatics analysis to predict the impact of the site on the protein structure,function and the signaling pathway where the protein is located.Results1.The clinic collects relevant clinical information of DPN patients,skin lesion manifestations,dermoscopic manifestations,and skin pathological characteristics.And make records,statistical summary of relevant results.2.DNA from skin lesions and peripheral blood of 10 DPN patients was extracted and sequenced by WES on the computer.The data were analyzed by bioinformatics software such as GATK,Annovar,Poly-Phen2,LRT,Mutation Taster and SIFT,including the quality of the sequencing data.Control,compare the reference genome,detect mutation sites,identify somatic mutations and germ cell mutations,annotate mutation sites,and then filter according to disease gene mutation-related databases such as Clin VAR,gnoma AD,OMMIN,etc.,and finally screen out all exons on DPN Regions of somatic potential pathogenic gene mutation sites.3.For the potential pathogenic gene mutation site,use bioinformatics analysis to predict the impact of the site on the protein structure,function and the signaling pathway where the protein is located.Conclusion1)In this study,through the clinical collection of case data,the distribution of DPN skin lesions,the dermoscopic definition and the characteristic manifestations of pathological sections were described in detail.It is helpful to reduce clinical misdiagnosis,to have a comprehensive and in-depth understanding of DPN,to improve the diagnosis rate of DPN,and to choose the right time for intervention.2)This paper reports for the first time the somatic gene mutation map of DPN patients in Asian population,revealing new gene mutation information of the disease.3)14 candidate DPN pathogenic genes including HRAS,CDC27,PHTF1,FGFR3,RBM12,CNTNAP5,HNMT,PHF5A,UBE2H,PTMA,DDX3X,CHIC1,PTEN and PUF60 were found by screening.Finally,a new non-synonymous mutation in HRAS gene was identified,and HRAS may be a potential pathogenic gene mutation of DPN.4)In this study,through bioinformatics analysis,it was found that DPN patients had abnormal signaling pathways such as EGFR.This also suggests that HRAS may be involved in the development of DPN.
Keywords/Search Tags:Dermatosis Papulosa Nigra, whole-exome sequencing, gene mutation, bioinformatics analysis, HRAS
PDF Full Text Request
Related items