Pedigree Analysis Of Primary Ciliary Dyskinesia Caused By Mutation In ODAD1 Gene

ObjectsIn this study,we analyzed the clinical manifestations,results of whale exone and sangex sequencing,splicing prediction software of members in a Chinese Han family with primary ciliary dyskinesia(PCD)to explore the genetic rules of PCD in this family.The pathogenic and mutaton site of the family were discovered,and thee pathogenicity of the mutation site was preliminarily verified so as to provide help for the clinical diagnosis and genetic comseling of the carrier.Methods1.The proband,her parents,siblings,daughter and other third generations of the family were selected as the study subjects.Their clinical information including age,gender,clinical symptoms,physical signs,imaging examinations was collected and the pedigree map was drawn.2.Peripheral blood samples were collected from most members of the family(12 members),and DNA extracted and subjected to whole ecome sequencing.3.WES and tollowed analysis indicated that several members of the family had matations of BDAD1 gene c.71-2A>C in q13.33 region of chromosome 19,among which the proband and his eldest brother were homozygous,and the proband’s mother,daughter and eldest brothers children were carriers.Combined with the clinical manitestations,the genotype and phenotype of the family members were separated,and this matation site was identified as the pathogenic site of PCD in the family.After checking dbSNP database,it was found that its rs number was 963154615,while 1000g2015aug_EAST esp6500siv2_EA5S ExAC EAS did not have this mutationAccording to ClinvarT OMIM and HGMD databases,no studies have been conducted on the pathogenicity of this mutation site and PCD.4.Sanger sequencing used to further verify the presence of the mutation in the proband.5.Hioinformatics software,included Human splicing finder(HSF)and Natgene2 server were used to predict the pathogenicity of this mutation,and the pathogenicity and related mechanisms were discussed based on the relevant information of the mutaiton site and the genetic rule in this family.Results1.The results of family investigation showed that the proband’s parents were inbred,their phenotype wss normal,and raised 1 son and 4 daughters.The proband and his eldest brother and second sister were PCD patients,all suffering from visceral inversion:bronchiectasis,and sinusitis,belonging to the PCD subtype Kartagener syndrome(KS).None of the third-generation individuals developed the disease.The genetic pattern of PCD in this family was determined to be antosomal recessive.2.WES and the results of data analysis indicated that several members of the family had a site mutation(c.71-2A>C)of ODAD 1 gene in q13.33 region of chromosome 19.The proband and her eldest brother were hamozygous,and the probands mother,daughter and eldest brother’s children were carners.Combined with the clinical manifestations,the genotype and phenotype of the family members were co-segregantion,and this site was identified as the pathogenic mutation of PCD in the family.After checking dbSNP database,we found that its rs number was 963 154615,while the mutation did not exist in 1000g2015aug_EAST esp6500siv2_EAS,or ExAC EAS.According to Clinvar,OMIM and HGMD databases,no studies have been conducted on the pathogenicity of this mutation site and PCD.Briefly,wefound a new mutation of PCD.3.The Sanger sequencing result verified the detection and confirmed that the proband carried the mutation.4.Human Splicing Finder saftware showed that the mutatian was located in the splice acceptor site of intron 3 of ODAD1 gene,which harmful and affected the splicing of precursor RNA.Netgence 2 server predicted that this mutation would cause the deletion of the splice receptor site in base no.1556 and affect the splicing of precursor RNA.Conclusions1.Several members of this family carry the mutation of ODAD1 gene c.71-2A>C in q13.33 regiom of chromosome 19,which is a new mutation site of PCD.2.The mutation is pathogenic and caused PCD by affecting the splicing of precursor RNA.