Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities...Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities.PCD is typically resulted from variants in genes encoding assembly or structural proteins that are indispensable for the movement of motile cilia.Here,we identified a novel nonsense mutation,c.466G>T,in cilia-and flagella-associated protein 300(CFAP300)resulting in a stop codon(p.Glu156*)through whole-exome sequencing(WES).The proband had a PCD phenotype with laterality defects and immotile sperm flagella displaying a combined loss of the inner dynein arm(IDA)and outer dynein arm(ODA).Bioinformatic programs predicted that the mutation is deleterious.Successful pregnancy was achieved through intracytoplasmic sperm injection(ICSI).Our results expand the spectrum of CFAP300 variants in PCD and provide reproductive guidance for infertile couples suffering from PCD caused by them.展开更多
Primary ciliary dyskinesia(PCD)is a genetically heterogeneous disorder characterized by impaired motility of cilia and flagella.Mutations in cilia-and flagella-associated protein 300(CFAP300)are associated with human ...Primary ciliary dyskinesia(PCD)is a genetically heterogeneous disorder characterized by impaired motility of cilia and flagella.Mutations in cilia-and flagella-associated protein 300(CFAP300)are associated with human PCD and male infertility;however,the underlying pathogenic mechanisms remain poorly understood.In a consanguineous Chinese family,we identified a homozygous CFAP300 loss-of-function variant(c.304delC)in a proband presenting with classical PCD symptoms and severe sperm abnormalities,including dynein arm deficiency and acrosomal malformation,as confirmed by transmission electron microscopy(TEM).Histological analysis revealed multiple morphological abnormalities of the sperm flagella in CFAP300-mutant individual,whereas immunofluorescence demonstrated markedly reduced CFAP300 expression in the spermatozoa of the proband.Furthermore,tandem mass tag(TMT)-based quantitative proteomics showed that the CFAP300 mutation reduced key spermatogenesis proteins(e.g.,sperm flagellar 2[SPEF2],solute carrier family 25 member 31[SLC25A31],and A-kinase anchoring protein 3[AKAP3])and mitochondrial ATP synthesis factors(e.g.,SLC25A31,cation channel sperm-associated 3[CATSPER3]).It also triggered abnormal increases in autophagy-related proteins and signaling mediator phosphorylation.These molecular alterations are likely to contribute to progressive deterioration of sperm ultrastructure and function.Notably,successful pregnancy was achieved via intracytoplasmic sperm injection(ICSI)using the proband’s sperm.Overall,this study expands the known CFAP300 mutational spectrum and offers novel mechanistic insights into its role in spermatogenesis.展开更多
基金supported by the China Postdoctoral Science Foundation Grant(2023M734294)Jiangsu Provincial Medical Key Discipline Cultivation Unit(JSDW202215)the National Natural Science Foundation of China(No.82001618).
文摘Primary ciliary dyskinesia(PCD)is a clinically rare,genetically and phenotypically heterogeneous condition characterized by chronic respiratory tract infections,male infertility,tympanitis,and laterality abnormalities.PCD is typically resulted from variants in genes encoding assembly or structural proteins that are indispensable for the movement of motile cilia.Here,we identified a novel nonsense mutation,c.466G>T,in cilia-and flagella-associated protein 300(CFAP300)resulting in a stop codon(p.Glu156*)through whole-exome sequencing(WES).The proband had a PCD phenotype with laterality defects and immotile sperm flagella displaying a combined loss of the inner dynein arm(IDA)and outer dynein arm(ODA).Bioinformatic programs predicted that the mutation is deleterious.Successful pregnancy was achieved through intracytoplasmic sperm injection(ICSI).Our results expand the spectrum of CFAP300 variants in PCD and provide reproductive guidance for infertile couples suffering from PCD caused by them.
基金supported by the National Natural Science Foundation of China(No.82101682)the Foundation of Education Department of Anhui Province for Outstanding Young Teachers(gxgwfx2022009).
文摘Primary ciliary dyskinesia(PCD)is a genetically heterogeneous disorder characterized by impaired motility of cilia and flagella.Mutations in cilia-and flagella-associated protein 300(CFAP300)are associated with human PCD and male infertility;however,the underlying pathogenic mechanisms remain poorly understood.In a consanguineous Chinese family,we identified a homozygous CFAP300 loss-of-function variant(c.304delC)in a proband presenting with classical PCD symptoms and severe sperm abnormalities,including dynein arm deficiency and acrosomal malformation,as confirmed by transmission electron microscopy(TEM).Histological analysis revealed multiple morphological abnormalities of the sperm flagella in CFAP300-mutant individual,whereas immunofluorescence demonstrated markedly reduced CFAP300 expression in the spermatozoa of the proband.Furthermore,tandem mass tag(TMT)-based quantitative proteomics showed that the CFAP300 mutation reduced key spermatogenesis proteins(e.g.,sperm flagellar 2[SPEF2],solute carrier family 25 member 31[SLC25A31],and A-kinase anchoring protein 3[AKAP3])and mitochondrial ATP synthesis factors(e.g.,SLC25A31,cation channel sperm-associated 3[CATSPER3]).It also triggered abnormal increases in autophagy-related proteins and signaling mediator phosphorylation.These molecular alterations are likely to contribute to progressive deterioration of sperm ultrastructure and function.Notably,successful pregnancy was achieved via intracytoplasmic sperm injection(ICSI)using the proband’s sperm.Overall,this study expands the known CFAP300 mutational spectrum and offers novel mechanistic insights into its role in spermatogenesis.
