Patients with multiple morphological abnormalities of the sperm flagella due to DNAH1 mutations have a good prognosis following intracytoplasmic sperm injection.
Does DNAH1 status influence intracytoplasmic sperm injection (ICSI) outcomes for patients with multiple morphological abnormalities of the sperm flagella (MMAF)?



Despite a highly abnormal morphology, sperm from MMAF patients with DNAH1 mutations have a low aneuploidy rate and good nuclear quality, leading to good embryonic development following ICSI and a high pregnancy rate.



Teratozoospermia represents a heterogeneous group including a wide range of phenotypes. Among all these qualitative defects, a flagellar phenotype called MMAF is characterized by a mosaic of morphological abnormalities of the flagellum, including coiled, bent, irregular, short or/and absent flagella, mainly due to the absence of the axonemal central pair microtubules. We previously demonstrated that homozygous mutations in the DNAH1 gene, encoding an inner arm heavy chain dynein, are frequently found in patients with MMAF (28% of the patients from the initial cohort). Numerous studies have reported an increased rate of aneuploidy and a poor sperm nuclear quality related to sperm flagellar abnormalities, which could impede ICSI outcome. Moreover, success rates after ICSI may be influenced by the type of ultrastructural flagellar defects and/or by the gene defects carried by the patients.



This retrospective cohort study included 6 infertile males with MMAF due to deleterious homozygous DNAH1 mutations and their respective spouses, who underwent 9 ISCI cycles, with 16 embryos being transferred. ICSI results were compared with two control populations of 13 MMAF men without DNAH1 mutations and an aged-matched control group of 1431 non-MMAF couples. All ICSI attempts took place between 2000 and 2012.



Clinical and biological data were collected from patients treated for infertility at the CPSR les Jasmins in Tunis (Tunisia). We compared the ICSI outcomes obtained with couples including DNAH1 mutated and nonmutated patients and non-MMAF couples. For the analysis of the chromosomal status, fluorescence in situ hybridization (FISH) analyses were performed on sperm cells from 3 DNAH1-mutated patients and from 29 fertile control subjects. Sperm chromatin condensation and DNA fragmentation were evaluated using aniline blue staining and TUNEL assays, respectively, on sperm cells from 3 DNAH1-mutated men and 6 fertile controls.



There was a significantly increased proportion of disomy XY and 18 in sperm from DNAH1 mutated patients compared with fertile controls (1.52 versus 0.28%, P = 0.0001 and 0.64 versus 0.09%, P = 0.0001). However, there were no statistically significant differences among sperm from the two groups in their frequencies of either 13, 21, XX or YY disomy or diploidy. Measures of DNA compaction and fragmentation demonstrated a good nuclear sperm quality among DNAH1 mutated men. The overall fertilization, pregnancy and delivery rates of couples including DNAH1 mutated men were of 70.8, 50.0 and 37.5%, respectively. There were no statistically significant differences in any of these parameters compared with the two control groups (P > 0.05).



A limitation of this study is the small number of DNAH1-mutated patients available and the low number of genes identified in MMAF. Further genetic studies are warranted to identify other MMAF-inducing genes to better characterize the genetic etiology of the MMAF phenotype and to improve the management of patients diagnosed with flagellar defects.



MMAF patients with DNAH1 mutations have low aneuploidy rates and good nuclear sperm quality, explaining the high pregnancy rate obtained with these patients. Good ICSI results were obtained for both MMAF groups (DNAH1 mutated and nonmutated), suggesting that patients presenting with asthenozoospermia due to flagellar defects have a good ICSI prognosis irrespective of their genotype. The majority of MMAF cases currently remain idiopathic with no genetic cause yet identified. In depth genetic analysis of these patients using next generation sequencing should reveal new causal genes. Subsequent genotype phenotype analyses could improve advice and care provided to MMAF patients.



None of the authors have any competing interest. This work is part of the project 'Identification and Characterization of Genes Involved in Infertility (ICG2I)', funded by the program GENOPAT 2009 from the French Research Agency (ANR) and the MAS-Flagella project, financed by the French ANR and the Direction Générale de l'Offre de Soins (DGOS).
Université Grenoble Alpes, Grenoble F-38000, France Equipe 'Genetics Epigenetics and Therapies of Infertility', Institut Albert Bonniot (IAB), INSERM U1209, CNRS UMR 5309, Grenoble F-38000, France CHU de Grenoble, UF de Génétique Chromosomique, Grenoble F-38000, France.
Clémentine
C
Wambergue
Polyclinique les Jasmins, Centre d'Aide Médicale à la Procréation, Centre Urbain Nord, 1003 Tunis, Tunisia.
Raoudha
R
Zouari
Polyclinique les Jasmins, Centre d'Aide Médicale à la Procréation, Centre Urbain Nord, 1003 Tunis, Tunisia.
Selima
S
Fourati Ben Mustapha
Université Grenoble Alpes, Grenoble F-38000, France Equipe 'Genetics Epigenetics and Therapies of Infertility', Institut Albert Bonniot (IAB), INSERM U1209, CNRS UMR 5309, Grenoble F-38000, France CHU de Grenoble, UF de Biologie de la procréation, Grenoble F-38000, France.
Guillaume
G
Martinez
CHU de Grenoble, UF de Génétique Chromosomique, Grenoble F-38000, France.
Françoise
F
Devillard
Université Grenoble Alpes, Grenoble F-38000, France Equipe 'Genetics Epigenetics and Therapies of Infertility', Institut Albert Bonniot (IAB), INSERM U1209, CNRS UMR 5309, Grenoble F-38000, France CHU de Grenoble, UF de Biologie de la procréation, Grenoble F-38000, France.
Sylviane
S
Hennebicq
Université Grenoble Alpes, Grenoble F-38000, France Equipe 'Genetics Epigenetics and Therapies of Infertility', Institut Albert Bonniot (IAB), INSERM U1209, CNRS UMR 5309, Grenoble F-38000, France CHU de Grenoble, UF de Génétique Chromosomique, Grenoble F-38000, France.
Véronique
V
Satre
Université Grenoble Alpes, Grenoble F-38000, France CHU de Grenoble, UF de Biologie de la procréation, Grenoble F-38000, France.
Sophie
S
Brouillet
Polyclinique les Jasmins, Centre d'Aide Médicale à la Procréation, Centre Urbain Nord, 1003 Tunis, Tunisia.
Lazhar
L
Halouani
Polyclinique les Jasmins, Centre d'Aide Médicale à la Procréation, Centre Urbain Nord, 1003 Tunis, Tunisia.
Ouafi
O
Marrakchi
Polyclinique les Jasmins, Centre d'Aide Médicale à la Procréation, Centre Urbain Nord, 1003 Tunis, Tunisia.
Mounir
M
Makni
Polyclinique les Jasmins, Centre d'Aide Médicale à la Procréation, Centre Urbain Nord, 1003 Tunis, Tunisia.
Habib
H
Latrous
Polyclinique les Jasmins, Centre d'Aide Médicale à la Procréation, Centre Urbain Nord, 1003 Tunis, Tunisia.
Mahmoud
M
Kharouf
CHU de Grenoble, UF de Génétique Chromosomique, Grenoble F-38000, France.
Florence
F
Amblard
Université Grenoble Alpes, Grenoble F-38000, France Equipe 'Genetics Epigenetics and Therapies of Infertility', Institut Albert Bonniot (IAB), INSERM U1209, CNRS UMR 5309, Grenoble F-38000, France.
Christophe
C
Arnoult
Université Grenoble Alpes, Grenoble F-38000, France Equipe 'Genetics Epigenetics and Therapies of Infertility', Institut Albert Bonniot (IAB), INSERM U1209, CNRS UMR 5309, Grenoble F-38000, France CHU de Grenoble, UF de Biochimie Génétique et Moléculaire, Grenoble F-38000, France pray@chu-grenoble.fr.
Pierre F
PF
Ray
Université Grenoble Alpes, Grenoble F-38000, France Equipe 'Genetics Epigenetics and Therapies of Infertility', Institut Albert Bonniot (IAB), INSERM U1209, CNRS UMR 5309, Grenoble F-38000, France CHU de Grenoble, UF de Génétique Chromosomique, Grenoble F-38000, France.
Charles
C
Coutton
D000328:Adult; D054468:Axoneme / Q000235:genetics* / Q000648:ultrastructure; D053938:DNA Fragmentation; D004398:Dyneins / Q000235:genetics*; D005260:Female; D005407:Flagella / Q000648:ultrastructure; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D020287:In Situ Nick-End Labeling; D007248:Infertility, Male / Q000235:genetics* / Q000628:therapy; D008297:Male; D009154:Mutation; D054315:Oocyte Retrieval; D010062:Ovulation Induction; D011247:Pregnancy; D018873:Pregnancy Rate; D011379:Prognosis; D012189:Retrospective Studies; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000002:abnormalities*; D016896:Treatment Outcome
C000598989:DNAH1 protein, human; D004398:Dyneins
DNAH1; FISH; ICSI; MMAF; flagellum; genetics; male infertility; teratozoospermia