Study On Etiology,Diagnosis And Treatment Of Infertility Caused By Primary Ciliary Dyskinesia

OBJECTIVE: To screen out the pathogenic genes associated with Primary ciliary dyskinesia?PCD?family,to clarify the differences in clinical manifestations between PCD family members,to demonstrate the correlation between severe asthenoteratozoospermia and sperm capacitation reduction in PCD patient,to apply assisted reproductive technology to treat infertility in PCD patient.CONTENTS: High-throughput gene microarrays sequencing was used to identify gene mutations in PCD patient and family members,the changes of cilia ultrastructure and different clinical manifestations between family members were observed.The expression of CatSper1,a key protein for sperm capacitation,was detected.Hypo-osmotic swelling test?HOST?was used to select sperm for intracytoplasmic sperm injection?ICSI?treatment.METHODS: Blood samples from PCD patients and family members were collected and detected for gene mutation by exon group capture and second generation sequencing,and the results were confirmed by Sanger sequencing.Sperm and bronchial cilia were collected and the ultrastructure was observed by transmission electron microscopy?TEM?.Respiratory function,CT,MRI and endoscopy were performed to identify the respiratory diseases of patients.Western blot was used to detect the expression of CatSper1 in the tail of sperm.Pre-genetic counseling was conducted for PCD infertility patient with severe teratozoospermia and a improved HOST was used to select sperm for ICSI treatment.RESULTS: 1.Severe sperm abnormalities were observed under optical microscopy,the tail was curled and enclosed in the plasma membrane.TEM showed the ultrastructure of outer dense fiber?ODF?,microtubules,radial spoke,outer dynein arm?ODA?and inner dynein arm?IDA?were gradually disordered from head to tail.2.The pulmonary function of the patients was obviously limited,with severe obstructive pulmonary ventilation dysfunction accompanied by diffuse dysfunction,severe small airway obstruction and severe decrease of pulmonary function.3.Imageology findings were as follows: paranasal sinusitis,mastoiditis,total visceral mirror transposition and bronchiectasis;Fiberoptic bronchoscopy showed extensivecongestion and edema of bronchial mucosa,increased secretion and inverted bronchoscopic image;TEM showed that the bronchial cilia were gradually disordered from distal to proximal segments,and there were significant differences in TEM expression among family members.4.Two previously unidentified suspected pathogenic mutations were detected in CCDC 40: c.1259 delA?p.Gln420 Glnfs X3;Het?and EX17₂0 DEL?Het?.5.Compared with normal semen and non-PCD asthenospermia,the expression of CatSper1 protein in proband was significantly down-regulated.CONCLUSION: 1.The novel mutations changed the sequence of CCDC40 protein,resulting in the ultrastructural defects of "9+2" cilia?ODF,microtubules,radial spoke,ODA and IDA are abnormal?,leading to the primary infertility in proband and a series of other PCD-related clinical manifestations.2.The abnormality of CCDC40 protein induced by the novel mutation caused a great change in sperm flagellum structure,which is the pathological basis of teratozoospermia of proband.3.The mutation of CCDC40 resulted in various ultrastructural phenotypes and great heterogeneity of cilia.There were also great heterogeneity in bronchoscopy,imageology and clinical manifestations.4.The expression of CatSper1 in proband was significantly down-regulated,suggesting that severe flagellar deformity may affect CatSper1 expression.5.Modified HOST+ICSI can achieve satisfactory clinical outcomes for PCD patients with severe infertility.