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Genetic Characterization of a Cohort of Italian Patients with Congenital Adrenal Hyperplasia Due to 21-Hydroxylase Deficiency

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Abstract

Introduction

Defects in the steroid 21-hydroxylase gene (CYP21A2) cause 21-hydroxylase deficiency (21OHD), the main cause of congenital adrenal hyperplasia (CAH). The disease shows a broad spectrum of clinical forms, ranging from severe or classical (salt wasting, SW, and simple virilizing, SV), to mild late onset or nonclassical (NC). 21OHD affects 1 in 15,000 in its severe classic form and 1 in 200–1000 in its mild NC form. There are many studies reporting the frequency of CYP21A2 pathogenic variants in different populations; however, few of them provide comprehensive information about Italian patients. Here, we present genetic results from a cohort of 245 unrelated Italian individuals with clinical diagnosis of CAH due to 21OHD.

Methods

A specific polymerase chain reaction (PCR) protocol combined with Sanger sequencing was used for CYP21A2 analysis. The multiplex ligation-dependent probe amplification (MLPA) assay was employed for copy number variation (CNV) determination.

Results

One hundred fourteen (46.5%) of the index cases had the NC form, 57 (23.3%) had the SV form, and 74 (30.2%) presented the SW form of the disease. The most prevalent variant found in NC patients was the p.Val282Leu (51.3%), while the most frequent variants in the classical form were p.Ile173Asn (8.6%) and c.293-13C>G (26.0%). In our study, the frequency of large rearrangements was 15.3%, with CAH-X alleles representing 40% of all DEL/CONV. In addition, 12 alleles carried rare variants, and 1 had a novel variant p.(Arg342Gln).

We observed phenotype-genotype correlation in 94.7% of cases. A complete concordance was observed in Groups 0 (enzyme activity completely impaired) where all patients had the SW form as expected. In Group A (0–1% residual enzyme activity), 78.4% of patients had the anticipated SW form while 21.6% were diagnosed with the SV form. Within Group B (~ 2% residual enzyme activity), 93.4% of patients exhibited SV form and 6.5% SW disease. Finally, 92.6% and 7.4% of patients belonging to Group C (enzyme partially impaired to ~ 20–60% residual activity) exhibited NC and SV phenotypes, respectively.

Conclusion

This work, representing a comprehensive genetic study, expanded the CYP21A2 variants spectrum of Italian patients with 21OHD and could be helpful in prenatal diagnosis and genetic counseling.

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Authors and Affiliations

  1. Clinical Chemistry, Biochemistry and Molecular Biology Operations (UOC), Agostino Gemelli Foundation University Hospital IRCCS, Largo A. Gemelli 8, 00168, Rome, Italy

    Paola Concolino, Alessia Perrucci, Cinzia Carrozza & Andrea Urbani

  2. Department of Basic Biotechnological Sciences, Intensivological and Perioperative Clinics, Catholic University of Sacred Heart, Rome, Italy

    Cinzia Carrozza & Andrea Urbani

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  1. Paola Concolino
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Correspondence to Paola Concolino.

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Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Conflict of Interest

PC, AP, CC, and AU have no conflicts of interest that are directly relevant to the content of this article.

Ethical Approval

Ethical review and approval were waived for this study, due to the diagnostic aim of the investigation. All procedures were performed in accordance with the ethical standards of the Ethics Committee of Fondazione Policlinico Universitario A. Gemelli IRCCS, Rome.

Informed Consent

Informed consent was obtained from all subjects involved in the study.

Availability of Data and Material

The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.

Authors’ Contributions

PC and AP performed the diagnostic investigations and prepared manuscript. CC and AU writing—review and editing. All authors have read and approved the final article.

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Concolino, P., Perrucci, A., Carrozza, C. et al. Genetic Characterization of a Cohort of Italian Patients with Congenital Adrenal Hyperplasia Due to 21-Hydroxylase Deficiency. Mol Diagn Ther 27, 621–630 (2023). https://doi.org/10.1007/s40291-023-00666-x

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