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
目的:探寻精子鞭毛多发形态异常(MMAF)可能的致病基因。方法:通过对1例典型的MMAF患者进行全外显子组测序(WES),分析可能的致病基因;运用扫描电镜和透射电镜观察MMAF患者精液样本,明确其鞭毛超微结构特点;通过精子免疫荧光技术分析cilia...目的:探寻精子鞭毛多发形态异常(MMAF)可能的致病基因。方法:通过对1例典型的MMAF患者进行全外显子组测序(WES),分析可能的致病基因;运用扫描电镜和透射电镜观察MMAF患者精液样本,明确其鞭毛超微结构特点;通过精子免疫荧光技术分析cilia and flagella-associated protein 65(CFAP65)在精子发生过程中的表达模式。结果:该例患者存在CFAP65基因的一个纯合致病性突变c.2675G>A(p.Trp892*);扫描电镜发现该患者精子具有典型的MMAF特征,即表现为无尾,折尾,卷尾,短尾或不规则尾巴;透射电镜发现患者精子鞭毛"9+2"结构缺失和紊乱:精子鞭毛纤维鞘组装异常,伴有中心微管缺失和动力蛋白臂缺失。细胞免疫荧光提示该CFAP65基因在小鼠各级生殖细胞均有表达。结论:CFAP65基因参与了精子鞭毛结构的组装,其突变可引起MMAF表型而导致男性不育。展开更多
As a specific type of asthenoteratozoospermia,multiple morphological abnormalities of the sperm flagella(MMAF)is characterized by composite abnormalities,including absent,short,coiled,angulation,and irregular-caliber ...As a specific type of asthenoteratozoospermia,multiple morphological abnormalities of the sperm flagella(MMAF)is characterized by composite abnormalities,including absent,short,coiled,angulation,and irregular-caliber flagella.Mutations in cilia-and flagella-associated protein 43(CFAP43)are one of the main causative factors of MMAF established to date.To identify whether there are other CFAP43 mutations related to MMAF and to determine the clinical outcomes of assisted reproductive technology for patients with MMAF harboring different mutations,we recruited and screened 30 MMAF-affected Chinese men using a 22-gene next-generation sequencing panel.After systematic analysis,seven mutations in CFAP43,including five novel mutations and two previously reported mutations,were identified from four families and related to MMAF in an autosomal recessive pattern.Papanicolaou staining,immunofluorescence,and electronic microscopy further clarified the semen characteristics a nd abnormal sperm morphologies,including disorganized axonemal and peri-axonemal structures,of the CFAP43-deficient men.The female partners of two patients were pregnant after undergoing assisted reproductive technology through intracytoplasmic sperm injection,and one of them successfully gave birth to a healthy boy.This study significantly expands the mutant spectrum of CFAP43,and together with the available information regarding male infertility and MMAF,provides new information for the genetic diagnosis and counseling of MMAF in the future.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
目的探讨CFAP43或CFAP44基因突变致精子鞭毛多发形态异常(multiple morphological abnormalities of the flagella,MMAF)患者行卵胞质内单精子注射(intracytoplasmic sperm injection,ICSI)助孕的临床结局。方法回顾性队列研究分析2014...目的探讨CFAP43或CFAP44基因突变致精子鞭毛多发形态异常(multiple morphological abnormalities of the flagella,MMAF)患者行卵胞质内单精子注射(intracytoplasmic sperm injection,ICSI)助孕的临床结局。方法回顾性队列研究分析2014年9月至2020年7月期间于安徽医科大学第一附属医院妇产科生殖医学中心就诊的121例MMAF男性不育症患者的临床资料和基因检测结果,纳入9例CFAP43或CFAP44基因突变MMAF患者,5例MMAF患者(P3、P5、P7、P8和P9)选择ICSI助孕治疗,统计并分析这5例患者ICSI助孕的临床结局。结果Sanger测序证实9例MMAF患者携带CFAP43或CFAP44基因双等位基因突变,其中3例患者的突变位点以往未曾报道,分别为CFAP43基因的新发纯合突变(c.4132delC:p.Arg1378Glufs*10)和新发复合杂合突变(c.3938G>A:p.Arg1313Gln;c.4342G>A:p.Glu1448Lys)以及CFAP44基因的新发复合杂合突变(c.1718C>A:p.Pro573His;c.4075G>A:p.Glu1359Lys)。5例MMAF患者夫妇接受5个ICSI周期,已生育4个健康亲生子代。CFAP43或CFAP44基因突变MMAF患者组ICSI受精率为76.47%(39/51),5例患者中临床妊娠3例,活产3例。与DNAH1基因突变MMAF患者组和严重少弱精子症患者组相比,CFAP43或CFAP44基因突变MMAF患者ICSI助孕结局差异均无统计学意义(均P>0.05)。结论CFAP43或CFAP44基因突变会导致精子严重的鞭毛畸形和运动能力下降,是MMAF的重要病因。ICSI技术可以有效地解决CFAP43或CFAP44基因突变MMAF患者的生育难题。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.
文摘目的:探寻精子鞭毛多发形态异常(MMAF)可能的致病基因。方法:通过对1例典型的MMAF患者进行全外显子组测序(WES),分析可能的致病基因;运用扫描电镜和透射电镜观察MMAF患者精液样本,明确其鞭毛超微结构特点;通过精子免疫荧光技术分析cilia and flagella-associated protein 65(CFAP65)在精子发生过程中的表达模式。结果:该例患者存在CFAP65基因的一个纯合致病性突变c.2675G>A(p.Trp892*);扫描电镜发现该患者精子具有典型的MMAF特征,即表现为无尾,折尾,卷尾,短尾或不规则尾巴;透射电镜发现患者精子鞭毛"9+2"结构缺失和紊乱:精子鞭毛纤维鞘组装异常,伴有中心微管缺失和动力蛋白臂缺失。细胞免疫荧光提示该CFAP65基因在小鼠各级生殖细胞均有表达。结论:CFAP65基因参与了精子鞭毛结构的组装,其突变可引起MMAF表型而导致男性不育。
基金the Natural Science Foundation of Chongqing(CSTC2021JCYJ-MSXMX0722)Chongqing Health Center for Women and Children(2020YJMS01).
文摘As a specific type of asthenoteratozoospermia,multiple morphological abnormalities of the sperm flagella(MMAF)is characterized by composite abnormalities,including absent,short,coiled,angulation,and irregular-caliber flagella.Mutations in cilia-and flagella-associated protein 43(CFAP43)are one of the main causative factors of MMAF established to date.To identify whether there are other CFAP43 mutations related to MMAF and to determine the clinical outcomes of assisted reproductive technology for patients with MMAF harboring different mutations,we recruited and screened 30 MMAF-affected Chinese men using a 22-gene next-generation sequencing panel.After systematic analysis,seven mutations in CFAP43,including five novel mutations and two previously reported mutations,were identified from four families and related to MMAF in an autosomal recessive pattern.Papanicolaou staining,immunofluorescence,and electronic microscopy further clarified the semen characteristics a nd abnormal sperm morphologies,including disorganized axonemal and peri-axonemal structures,of the CFAP43-deficient men.The female partners of two patients were pregnant after undergoing assisted reproductive technology through intracytoplasmic sperm injection,and one of them successfully gave birth to a healthy boy.This study significantly expands the mutant spectrum of CFAP43,and together with the available information regarding male infertility and MMAF,provides new information for the genetic diagnosis and counseling of MMAF in the future.
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.
文摘目的探讨CFAP43或CFAP44基因突变致精子鞭毛多发形态异常(multiple morphological abnormalities of the flagella,MMAF)患者行卵胞质内单精子注射(intracytoplasmic sperm injection,ICSI)助孕的临床结局。方法回顾性队列研究分析2014年9月至2020年7月期间于安徽医科大学第一附属医院妇产科生殖医学中心就诊的121例MMAF男性不育症患者的临床资料和基因检测结果,纳入9例CFAP43或CFAP44基因突变MMAF患者,5例MMAF患者(P3、P5、P7、P8和P9)选择ICSI助孕治疗,统计并分析这5例患者ICSI助孕的临床结局。结果Sanger测序证实9例MMAF患者携带CFAP43或CFAP44基因双等位基因突变,其中3例患者的突变位点以往未曾报道,分别为CFAP43基因的新发纯合突变(c.4132delC:p.Arg1378Glufs*10)和新发复合杂合突变(c.3938G>A:p.Arg1313Gln;c.4342G>A:p.Glu1448Lys)以及CFAP44基因的新发复合杂合突变(c.1718C>A:p.Pro573His;c.4075G>A:p.Glu1359Lys)。5例MMAF患者夫妇接受5个ICSI周期,已生育4个健康亲生子代。CFAP43或CFAP44基因突变MMAF患者组ICSI受精率为76.47%(39/51),5例患者中临床妊娠3例,活产3例。与DNAH1基因突变MMAF患者组和严重少弱精子症患者组相比,CFAP43或CFAP44基因突变MMAF患者ICSI助孕结局差异均无统计学意义(均P>0.05)。结论CFAP43或CFAP44基因突变会导致精子严重的鞭毛畸形和运动能力下降,是MMAF的重要病因。ICSI技术可以有效地解决CFAP43或CFAP44基因突变MMAF患者的生育难题。
