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The treatment of obstructive azoospermia by intracytoplasmic sperm injection

Traitement de l’azoospermie obstructive par injection Intracytoplasmique d’un spermatozoïde

Abstract

Intracytoplasmic sperm injection (ICSI) allows the treatment of virtually every type of male infertility. Unlike in vitro fertilization (IVF), its success does not depend on sperm concentration, motility or morphology and most of the physical barriers to fertilisation are by-passes. Since ICSI does not require strongly motile sperm, its use has now been expanded to incorporate immature sperm from the testes and epididymides. Successful fertilisation, pregnancies and healthy babies have all been reported. However, concerns about the safety of ICSI remain due to its short clinical history and the lack of testing on animal models.

Male fertility potential for assisted reproduction by ICSI cannot be measured by conventional parameters. Sperm DNA integrity is increasingly recognised as a more useful indicator. Studies have shown that sperm with higher levels of DNA damage have lower fertilisation rates after IVF and ICSI. Sperm with DNA damage above a certain threshold are associated with a longer time to conceive in otherwise apparently fertile couples and a higher miscarriage rate. DNA damage has been shown to be associated with impaired embryo cleavage. Our group has shown that sperm DNA from testicular sperm is less fragmented than that from epididymal sperm and suggest its preferred use in ICSI.

In addition to nuclear (n) DNA we also assessed the quality of mitochondrial (mt) DNA from testicular sperm from men with obstructive azoospermia undergoing ICSI. We observed that couples achieving a pregnancy had both less mtDNA deletions and less nDNA fragmentation. We found inverse relationships between pregnancy and sperm mtDNA deletion numbers, size and nDNA fragmentation. No relationships were observed with fertilisation rates. With this knowledge, we designed an algorithm for the prediction of pregnancy based on the quality of sperm nDNA and mtDNA.

Each year 40,000 men have a vasectomy in the UK but every year 2500 request a reversal to begin a second family. For such men, vasectomy reversal has recently been replaced in part by testicular biopsy via fine-needle testicular sperm aspiration (TESA) or percutaneous epididymal sperm aspiration (PESA) performed at an outpatient clinic and subsequently used in ICSI. Since these were previously fertile men it has been assumed that they had ‘fertile’ sperm. However the assited conception success rates of these mens partners has not been assessed until recently. We have shown a significant reduction in the clinical pregnancy rates in the partners of men who had had a vasectomy ≥10yrs previously. There is also evidence to suggest that spermatogenesis is significantly impaired in vasectomised men. Marked decreases in spermatocytes, spermatids and spermatozoa have been observed. We have found this to be associated with concomitant increases in apoptotic markers, such as Fas, FasL and Bax. The quality of the remaining sperm is also compromised. Sperm DNA from vasectomized men shows substantial damage which increases with time after surgery. This new use of ICSI will be discussed.

Resume

L’injection intracytoplasmique d’un spermatozoïde (ICSI) permet de traiter virtuellement n’importe quel type d’infécondité masculine. A la différence de la Fécondation in vitro (FIV), son succès ne dépend pas de la concentration, de la mobilité ou de la morphologie des spermatozoïdes et la plupart des barrières physiques à la fécondation sont ainsi contournées. Puisque l’ICSI ne requiert pas de spermatozoïdes fortement mobiles, son usage a été maintenant élargi à l’utilisation des spermatozoïdes immatures des testicules et des épididymes. Une fécondation réussie, des grossesses, et des enfants en bonne santé ont tous été rapportés. Toutefois, persistent des inquiétudes à propos de l’innocuité de l’ICSI qui sont dues à sa brève existence clinique et à l’absence d’expérimentation sur des modèles animaux.

Le potentiel de fécondité masculine pour la reproduction assistée par ICSI ne peut pas être mesuré par les paramètres conventionnels. L’intégrité de l’ADN spermatique est, de façon grandissante, reconnue comme un indicateur plus utile. Des études ont montré que les spermatozoïdes ayant des taux élevés d’altération de l’ADN, ont des taux de fécondation plus faibles en FIV et en ICSI. Les spermatozoïdes avec une altération de l’ADN au-dessus d’un certain seuil sont associés à une augmentation du délai pour concevoir chez des couples par ailleurs apparemment féconds, et à un taux élevé de fausses-couches. On a montré que l’altération de l’ADN était liée à une segmentation embryonnaire défectueuse. Notre groupe a rapporté que l’ADN de spermatozoïdes provenant du testicule était moins fragmenté que l’ADN de spermatozoïdes provenant de l’épididyme, ce qui suggère l’utilisation préférée des premiers en ICSI.

En plus de l’ADN nucléaire (n), nous avons aussi évalué la qualité de l’ADN mitochondrial (mt) dans les spermatozoïdes testiculaires d’hommes avec azoospermie obstructive en cours d’ICSI.

Nous avons observé que les couples qui obtenaient une grossesse, avaient à la fois moins de délétions de l’ADNmt et moins de fragmentation de l’ADNn. Nous avons trouvé une relation inverse entre grossesse d’une part, et d’autre part, nombres de délétion de l’ADNmet et taille et fragmentation de l’ADNn des spermatozoïdes. Aucune relation n’a été observée avec les taux de fécondation. Avec ces données, nous avons établi un algorithme pour la prédiction de grossesses, basé sur la qualité de l’ADN, et l’ADNmt des spermatozoïdes.

Chaque année 40 000 hommes ont une vasectomie au Royaume Uni, mais chaque année 2500 demandent une reperméabilisation pour fonder une seconde famille. Pour de tels hommes, la réversion de la vasectomie a récemment été remplacée en partie par la biopsie testiculaire, au moyen de l’aspiration de spermatozoïdes testiculaires à l’aiguille fine (TESA) ou de l’aspiration percutanée de spermatozoïdes épididymaires (PESA), réalisée en ambulatoire et utilisée ensuite en ICSI. Puisque ces hommes étaient auparavant des hommes féconds, on les a considérés comme ayant des spermatozoïdes “féconds”. Toutefois, les taux de succès en conception assistée des partenaires de ces hommes n’ont été évalués que récemment. Nous avons montré une réduction significative des taux de grossesse clinique chez les partenaires des hommes qui avaient eu une vasectomie plus de 10 ans auparavant. II existe aussi des éléments qui suggèrent que la spermatogenèse est altérée de façon significative chez les hommes vasectomisés. Une diminution importante des spermatocytes, des spermatides et des spermatozoïdes a été observée. Nous avons trouvé que celle-ci était associée à une augmentation concomitante des marqueurs de l’apoptose, comme le Fas, FasL et Bax. La qualité des spermatozoïdes qui restent est aussi compromise. L’ADN des spermatozoïdes d’hommes vasectomisés présente des altérations substantielles qui augmentent avec le temps après la chirurgie.

L’auteur discute cette nouvelle indication de l’ICSI.

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Correspondence to Sheena E. M. Lewis.

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Lewis, S.E.M. The treatment of obstructive azoospermia by intracytoplasmic sperm injection. Androl. 16, 28–38 (2006). https://doi.org/10.1007/BF03034829

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