Clinical Evaluation of a Custom Gene Panel as a Tool for Precision Male Infertility Diagnosis by Next-Generation Sequencing

Cannarella, Rossella; Precone, Vincenza; Busetto, Gian Maria; Renzo, Gian Carlo Di; Gerli, Sandro; Manara, Elena; Dautaj, Astrit; Bertelli, Matteo; Calogero, Aldo E.

doi:10.3390/life10100242

Cited by 17 publications

(8 citation statements)

References 59 publications

(73 reference statements)

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“…The FLRT3 variant found in Patient 4 has already been described in the literature as a rare missense variant of unknown association with CHARGE syndrome [ 41 ]. The CHD7 variant, already described by our group [ 42 ], is a VUS of difficult interpretation, but molecular modeling suggests a potential pathogenic role. If it were true, Patient 4 would be the first case of reversal cHH associated with a CHD7 variant.…”

Section: Discussionmentioning

confidence: 99%

Genetic Analysis of Patients with Congenital Hypogonadotropic Hypogonadism: A Case Series

Cannarella

Gusmano

Condorelli

et al. 2023

IJMS

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Congenital hypogonadotropic hypogonadism (cHH)/Kallmann syndrome (KS) is a rare genetic disorder with variable penetrance and a complex inheritance pattern. Consequently, it does not always follow Mendelian laws. More recently, digenic and oligogenic transmission has been recognized in 1.5–15% of cases. We report the results of a clinical and genetic investigation of five unrelated patients with cHH/KS analyzed using a customized gene panel. Patients were diagnosed according to the clinical, hormonal, and radiological criteria of the European Consensus Statement. DNA was analyzed using next-generation sequencing with a customized panel that included 31 genes. When available, first-degree relatives of the probands were also analyzed to assess genotype–phenotype segregation. The consequences of the identified variants on gene function were evaluated by analyzing the conservation of amino acids across species and by using molecular modeling. We found one new pathogenic variant of the CHD7 gene (c.576T>A, p.Tyr1928) and three new variants of unknown significance (VUSs) in IL17RD (c.960G>A, p.Met320Ile), FGF17 (c.208G>A, p.Gly70Arg), and DUSP6 (c.434T>G, p.Leu145Arg). All were present in the heterozygous state. Previously reported heterozygous variants were also found in the PROK2 (c.163del, p.Ile55*), CHD7 (c.c.2750C>T, p.Thr917Met and c.7891C>T, p.Arg2631*), FLRT3 (c.1106C>T, p.Ala369Val), and CCDC103 (c.461A>C, p.His154Pro) genes. Molecular modeling, molecular dynamics, and conservation analyses were performed on three out of the nine variants identified in our patients, namely, FGF17 (p.Gly70Arg), DUSP6 (p.Leu145Arg), and CHD7 p.(Thr917Met). Except for DUSP6, where the L145R variant was shown to disrupt the interaction between β6 and β3, needed for extracellular signal-regulated kinase 2 (ERK2) binding and recognition, no significant changes were identified between the wild-types and mutants of the other proteins. We found a new pathogenic variant of the CHD7 gene. The molecular modeling results suggest that the VUS of the DUSP6 (c.434T>G, p.Leu145Arg) gene may play a role in the pathogenesis of cHH. However, our analysis indicates that it is unlikely that the VUSs for the IL17RD (c.960G>A, p.Met320Ile) and FGF17 (c.208G>A, p.Gly70Arg) genes are involved in the pathogenesis of cHH. Functional studies are needed to confirm this hypothesis.

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Section: Discussionmentioning

confidence: 99%

Genetic Analysis of Patients with Congenital Hypogonadotropic Hypogonadism: A Case Series

Cannarella

Gusmano

Condorelli

et al. 2023

IJMS

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“…One can expect that next-generation sequencing (NGS)-based approaches might facilitate this task. Although NGS analysis of custom gene panels was able to identify genetic events possibly involved in idiopathic male infertility development [ 24 , 25 ], the majority of the cases remained unexplained. This suggests the possible involvement of other, non-genetic, factors in the pathogenesis of idiopathic male infertility.…”

Section: Accumulating Evidence Points To the Involvement Of Ssc Niche...mentioning

confidence: 99%

Regenerative medicine for male infertility: A focus on stem cell niche injury models

Sagaradze¹,

Monakova²,

Basalova³

et al. 2022

Biomedical Journal

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“…Sanger sequencing was performed when target region coverage was less than 10 reads. All laboratory methods have been described elsewhere [11]. We searched international databases dbSNP (www.ncbi.nlm.nih.…”

Section: Bioinformatics and Genetic Analysismentioning

confidence: 99%

Analysis of 29 Targeted Genes for Non-obstructive Azoospermia: the Relationship Between Genetic Testing and Testicular Histology

Cannarella

Bertelli²,

Condorelli

et al. 2021

Preprint

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BACKGROUND. Few studies have evaluated the relationship between testicular histology and pathogenic variations of genes regulating spermatogenesis.AIM. To analyze the presence of potentially pathogenic variants of 29 candidate genes known to cause spermatogenic failure (SPGF) in patients with non-obstructive azoospermia (NOA) who underwent testicular histology.PATIENTS AND METHODS. Sixty patients with NOA referred to the Department of Transfusion Medicine and Transplantation Biology, University Hospital Center Zagreb, Croatia, for testicular biopsy were consecutively assessed for eligibility. Twelve patients were excluded from the study because they had Klinefelter syndrome (n=1), Yq microdeletions (n=6), testicular trauma (n=2), or in-situ germ cell neoplasia (n=3). Therefore, 48 patients were considered eligible and included in this study. They were divided into three groups: those who had cryptorchidism (n=9), those with varicocele (n=14), and those with idiopathic NOA (n=25). All included patients underwent blood withdrawal for next-generation sequencing analysis and gene sequencing.RESULTS. We found a possible genetic cause in 4 patients with idiopathic NOA (16%) and in 2 with cryptorchidism (22%). No pathogenic or possibly pathogenic mutations were identified in patients with varicocele. Variants of undetermined significance (VUS) were found in 11 patients with idiopathic NOA (44%), 3 with cryptorchidism (33%), and 8 patients with varicocele (57%). VUSs of the USP9Y gene were the most frequently as they were found in 14 out of 48 patients (29%). In particular, the VUS USP9Y c.7434+14del was found in 11 patients. They showed varied histological pictures, including Sertoli cell-only syndrome, mixed atrophy, and hypospermatogenesis, regardless of cryptorchidism or varicocele. No direct correlation was found between the gene mutation/variant and the testicular histological picture. CONCLUSION. Different mutations of the same gene cause various testicular histological pictures. These results suggest that it is not the gene itself but the type of mutation/variation that determines the testicular histology picture. Based on the data presented above, it remains challenging to design a genetic panel with prognostic value for the outcome of testicular sperm extraction in patients with NOA.

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Clinical Evaluation of a Custom Gene Panel as a Tool for Precision Male Infertility Diagnosis by Next-Generation Sequencing

Cited by 17 publications

References 59 publications

Genetic Analysis of Patients with Congenital Hypogonadotropic Hypogonadism: A Case Series

Genetic Analysis of Patients with Congenital Hypogonadotropic Hypogonadism: A Case Series

Regenerative medicine for male infertility: A focus on stem cell niche injury models

Analysis of 29 Targeted Genes for Non-obstructive Azoospermia: the Relationship Between Genetic Testing and Testicular Histology

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