Özgün Bir Genetik Protokol İle Erkek İnfertil Hastalarda Y Mikrodelesyonlarının İncelenmesi

Yıl 2025, Cilt: 47 Sayı: 1, 110 - 127, 17.01.2025

Aysen Akhundova , Sezin Canbek , Burcu Turkgenc

Öz

Düzenli, korunmasız vajinal cinsel ilişkiden 12 ay veya daha uzun süre sonra gebe kalamama, infertilite olarak bilinmektedir. Oligozospermi ve azospermiye bağlı erkek infertilitesi tanılı hastaların %30'unda infertilite, genetik nedenlerle ilişkilidir. İdiyopatik azospermide (%15-20) ve idiyopatik oligozoospermide (%7-10), Y kromozom mikrodelesyonları görülmektedir. Komple veya parsiyel delesyon durumu, hastanın canlı sperm üretimini etkilemektedir. Çalışmamızda, EAA/EMQN 2023 klavuzu önerileri dikkate alınarak, özgün bir genetik protokol ile erkek infertil hastalarda parsiyel ve komple Y mikrodelesyonlarının incelenmesi amaçlandı. Çalışmada çoğu azospermili, 45 hastaya ait DNA materyali, 1 sağlıklı kadın ve 1 sağlıklı erkek kontrol DNA’sı, Y kromozomu AZF komple ve parsiyel mikrodelesyon tayinini gerçekleştirmek üzere, altın standart olarak görülen iki aşamalı multipleks PZR metodolojisini kullanıldı. Çalışmamızda erkek infertilitesi tanısı almış 45 erkek hastada Y mikrodelesyon oranı %11,1 olarak belirlenmiştir. Delesyon belirlenen toplam 5 hastanın 2’sinde (Hasta 21 ve 31), Y kromozomu terminal heterokromatin bölgenin (Sy160) de kaybıyla eşlik eden parsiyel AZFb+c (%40), 1’inde (Hasta 23) Y kromozomu terminal heterokromatin bölgenin (Sy160) korunduğu komple AZFc (%20), 2 hastada (Hasta 25 ve 35) ise komple AZFa+b+c (%40) mikrodelesyonu saptanmıştır. Komple AZFa+b+c delesyonları saptanan Hasta 25 ve Hasta 35’de ZFY/ZFX ve SRY (sY14) pozitif olarak değerlendirilmiştir. Hasta 35’in karyotip analiz sonucu 46,XX erkek olarak tespit edilmiştir. Güncel 2023 EAA/EMQN kılavuzları dikkate alınarak geliştirdiğimiz benzersiz genetik protokol ile, erkek infertil hastalarda, Y mikrodelesyonlarının yüksek hassasiyet ve doğrulukla incelenmesi sağlanmıştır ve önceki protokollere göre avantajlar sağlamaktadır. AZF komple ve parsiyel delesyonların tanısı prognostik öneme sahiptir ve hastanın tedavi seçeneklerini büyük oranda etkiler.

Anahtar Kelimeler

Azospermi, AZF, erkek infertilitesi, Y kromozom mikrodelesyon, multipleks PZR

Etik Beyan

Etik Kurul Onayı: Çalışma Üsküdar Üniversitesi Girişimsel Olmayan Araştırmalar Etik Kurulu Başkanlığı tarafından onaylanmıştır (Karar no: 61351342/ OCAK 2024-96, Tarih: 31.01.2024). Onam: Yazarlar, çalışmaya dahil edilen tüm gönüllülerden bilgilendirilmiş onam alındığını beyan etmişlerdir.

Proje Numarası

No project grant was received for the study.

Teşekkür

Bu çalışma, Aysen Akhundova’nın yüksek lisans tez çalışması olarak Burcu Türkgenç danışmanlığında gerçekleştirilmiştir. Laboratuvar deneyleri, Üsküdar Üniversitesi Tıp Fakültesi Tıbbi Genetik Labı’nda geçekleştirilmiş olup lab sorumluları Prof. Dr. Muhsin Konuk ve Lab. Uzm. Tayfun Gözler’e, laboratuvarı kullanımımıza açtıkları ve ihtiyacımız olan eksik malzemelerin temininde desteklerinden ötürü çok teşekkür ederiz.

An Original Genetic Protocol For The Investigation of Y Microdeletions In Male Infertile Patients

Öz

The inability to conceive during 12 months or more of consistent, unprotected vaginal sex is known as infertility. In 30% of patients diagnosed with male infertility due to oligozoospermia and azoospermia, infertility is related to genetic causes. Y chromosome microdeletions are seen in idiopathic azoospermia (15-20%) and idiopathic oligozoospermia (7-10%). Complete or partial deletion status affects the patient's production of viable sperm. In this study we aimed to investigate partial and complete Y microdeletions in male infertile patients with a unique genetic protocol, taking into account the recommendations of the EAA/EMQN 2023 guidelines. In this study, DNA material from 45 patients, most of whom had azoospermia, 1 healthy female, and 1 healthy male control DNA were analyzed for Y chromosome AZF complete and partial microdeletions using the gold standard two-step multiplex PCR methodology. In our study, the Y microdeletion rate was 11.1% in 45 male patients diagnosed with male infertility. Of a total of 5 patients with deletions, 2 (Patients 21 and 31) had partial AZFb+c (40%) with loss of the Y chromosome terminal heterochromatin region (Sy160), 1 (Patient 23) had complete AZFc (20%) with preservation of the Y chromosome terminal heterochromatin region (Sy160), and 2 (Patients 25 and 35) had complete AZFa+b+c (40%) microdeletions. ZFY/ZFX and SRY (sY14) were preserved in Patient 25 and Patient 35 with complete AZFa+b+c deletions. The karyotype analysis result of Patient 35 was 46, XX male. A unique genetic protocol, based on the current 2023 EAA/EMQN guidelines, has been developed to examine Y microdeletions in male infertile patients with high sensitivity and accuracy and provides advantages over previous protocols.

Anahtar Kelimeler

Azoospermia, AZF, male infertility, Y chromosome microdeletion, multiplex PCR

Etik Beyan

Ethics Committee Approval: The study was approved by the Üsküdar University Non-Interventional Research Ethics Committee (Decision no: 61351342/ OCAK 2024-96, Date: 31.01.2024). Informed Consent: The authors declared that informed consent was obtained from all volunteers included in the study.

Destekleyen Kurum

Uskudar University, Medical Faculty, Medical Genetics laboratory

Proje Numarası

No project grant was received for the study.

Teşekkür

This study was conducted under the supervision of Burcu Turkgenc as Aysen Akhundova's master's thesis. Laboratory experiments were carried out in the Uskudar University Faculty of Medicine Medical Genetics Laboratory and we would like to thank Prof. Dr. Muhsin Konuk and Lab Specialist Tayfun Gözler for opening the laboratory for our use and for their support in providing the required materials we needed.

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