https://doi.org/10.4081/aiua.2024.12464
CFTR Exon 10 deleterious mutations in patients with congenital bilateral absence of vas deferens in a cohort of Pakistani patients
Submitted: March 16, 2024
Accepted: April 5, 2024
Published: October 2, 2024
Accepted: April 5, 2024
Abstract Views: 1236
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Supplementary: 152
Supplementary: 152
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All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Authors
Congenital bilateral absence of vas deferens (CBAVD) is a urological syndrome of Wolffian ducts and is responsible for male infertility and obstructive azoospermia. This study is designed to explore the integrity of exon 10 of CFTR and its role in male infertility in a cohort of CBVAD patients in Pakistan. Genomic DNA was extracted from 17 male patients with CBAVD having clinical symptoms, and 10 healthy controls via phenol-chloroform method. Exon 10 of the CFTR gene was amplified, using PCR with specific primers and DNA screening was done by Sanger sequencing. Sequencing results were analyzed using freeware Serial Cloner, SnapGene, BioEdit and FinchTV. Furthermore, bioinformatics tools were used to analyze the mutations and their impact on the protein function and stability. We have identified 4 mutations on exon 10 of CFTR in 6 out of 17 patients. Two of the mutations were missense variants V456A, K464E, and the other two were silent mutations G437G, S431S. The identified variant V456A was present in 4 of the studied patients. Whereas, the presence of K464E in our patients further weighs on the crucial importance for its strategic location to influence the gene function at post-transcriptional and protein level. Furthermore, Polyphen-2 and SIFT analyze the mutations as harmful and deleterious. The recurrence of V456A and tactically conserved locality of K464E are evidence of their potential role in CBAVD patients and in male infertility. The data can contribute in developing genetic testing and treatment of CBAVD.
Cai H, Qing X, Niringiyumukiza JD, et al. CFTR variants and renal abnormalities in males with congenital unilateral absence of the vas deferens (CUAVD): a systematic review and meta-analysis of observational studies. Genet Med. 2019;21:826-36.
DOI: https://doi.org/10.1038/s41436-018-0262-7
Halder A, Pandey D. CFTR gene variants in Indian congenital bilateral absence of vas deferens & its relevance in genetic counselling. Indian J Med Res. 2020;152:535-7.
DOI: https://doi.org/10.4103/ijmr.IJMR_649_20
Cai Z, Li H. Congenital bilateral absence of the vas deferens. Front Genet. 2022;13:775123.
DOI: https://doi.org/10.3389/fgene.2022.775123
Ferlin A, Dipresa S, Delbarba A, et al. Contemporary genetics-based diagnostics of male infertility. Expert Rev Mol Diagn. 2019;19:623-33.
DOI: https://doi.org/10.1080/14737159.2019.1633917
Daudin M, Bieth E, Bujan L, et al. Congenital bilateral absence of the vas deferens: clinical characteristics, biological parameters, cystic fibrosis transmembrane conductance regulator gene mutations, and implications for genetic counseling. Fertil Steril. 2000;74:1164-74.
DOI: https://doi.org/10.1016/S0015-0282(00)01625-3
Mieusset R, Bieth E, Daudin M, et al. Male partners of infertile couples with congenital unilateral absence of the vas deferens are mainly non-azoospermic. Andrology. 2020;8:645-53.
DOI: https://doi.org/10.1111/andr.12749
Bieth E, Hamdi SM, Mieusset R. Genetics of the congenital absence of the vas deferens. Hum Genet. 2021;140:59-76.
DOI: https://doi.org/10.1007/s00439-020-02122-w
Casals T, Bassas L, Egozcue S, et al. Heterogeneity for mutations in the CFTR gene and clinical correlations in patients with congenital absence of the vas deferens. Hum Reprod. 2000;15:1476-83.
DOI: https://doi.org/10.1093/humrep/15.7.1476
Akinsal EC, Baydilli N, Dogan ME, Ekmekcioglu O. Comorbidity of the congenital absence of the vas deferens. Andrologia. 2018;50:e12994.
DOI: https://doi.org/10.1111/and.12994
Li C-Y, Jiang L-Y, Chen W-Y, et al. CFTR is essential for sperm fertilizing capacity and is correlated with sperm quality in humans. Human Reprod. 2010;25:317-27.
DOI: https://doi.org/10.1093/humrep/dep406
Gaillard DA, Carre-Pigeon F, Lallemand A. Normal vas deferens in fetuses with cystic fibrosis. J Urol. 1997;158:1549-52.
DOI: https://doi.org/10.1016/S0022-5347(01)64278-2
Hwang TC, Yeh JT, Zhang J, et al. Structural mechanisms of CFTR function and dysfunction. J Gen Physiol. 2018;150:539-70.
DOI: https://doi.org/10.1085/jgp.201711946
Shishido H, Yoon JS, Yang Z, Skach WR. CFTR trafficking mutations disrupt cotranslational protein folding by targeting biosynthetic intermediates. Nat Commun. 2020;11:4258.
DOI: https://doi.org/10.1038/s41467-020-18101-8
Morris-Rosendahl DJ, Edwards M, McDonnell MJ, et al. Whole-gene sequencing of CFTR reveals a high prevalence of the intronic variant c.3874-4522A>G in cystic fibrosis. Am J Respir Crit Care Med. 2020;201:1438-41.
DOI: https://doi.org/10.1164/rccm.201908-1541LE
Indika NLR, Vidanapathirana DM, Dilanthi HW, et al. Phenotypic spectrum and genetic heterogeneity of cystic fibrosis in Sri Lanka. BMC Med Genet. 2019;20:89.
DOI: https://doi.org/10.1186/s12881-019-0815-x
Ferlin A, Stuppia L. Diagnostics of CFTR-negative patients with congenital bilateral absence of vas deferens: which mutations are of most interest? Expert Rev Mol Diagn. 2020;20:265-7.
DOI: https://doi.org/10.1080/14737159.2020.1707081
Gupta N, Sarkar S, Mehta P, et al. Polymorphisms in the HSF2, LRRC6, MEIG1 and PTIP genes correlate with sperm motility in idiopathic infertility. Andrologia. 2022;54:e14517.
DOI: https://doi.org/10.1111/and.14517
Ashkenazy H, Abadi S, Martz E, et al. ConSurf 2016: an improved methodology to estimate and visualize evolutionary conservation in macromolecules. Nucleic Acids Res. 2016;44:W344-50.
DOI: https://doi.org/10.1093/nar/gkw408
Venselaar H, Te Beek TA, Kuipers RK, et al. Protein structure analysis of mutations causing inheritable diseases. An e-Science approach with life scientist friendly interfaces. BMC Bioinformatics. 2010;11:548.
DOI: https://doi.org/10.1186/1471-2105-11-548
Giordano SH. Breast cancer in men. N Engl J Med. 2018;378:2311-20.
DOI: https://doi.org/10.1056/NEJMra1707939
Flanagan SE, Patch AM, Ellard S. Using SIFT and PolyPhen to predict loss-of-function and gain-of-function mutations. Genet Test Mol Biomarkers. 2010;14:533-7.
DOI: https://doi.org/10.1089/gtmb.2010.0036
Soegaard M, Kjaer SK, Cox M, et al. BRCA1 and BRCA2 mutation prevalence and clinical characteristics of a population-based series of ovarian cancer cases from Denmark. Clin Cancer Res. 2008;14:3761-7.
DOI: https://doi.org/10.1158/1078-0432.CCR-07-4806
Adzhubei I, Jordan DM, Sunyaev SR. Predicting functional effect of human missense mutations using PolyPhen-2. Curr Protoc Hum Genet. 2013;Chapter 7:Unit7.20.
DOI: https://doi.org/10.1002/0471142905.hg0720s76
Pulumati A, Pulumati A, Dwarakanath BS, et al. Technological advancements in cancer diagnostics: Improvements and limitations. Cancer Rep (Hoboken). 2023;6:e1764.
DOI: https://doi.org/10.1002/cnr2.1764
Pires DE, Ascher DB, Blundell TL. mCSM: predicting the effects of mutations in proteins using graph-based signatures. Bioinformatics. 2014;30:335-42.
DOI: https://doi.org/10.1093/bioinformatics/btt691
Lewis HA, Zhao X, Wang C, et al. Impact of the deltaF508 mutation in first nucleotide-binding domain of human cystic fibrosis transmembrane conductance regulator on domain folding and structure. J Biol Chem. 2005;280:1346-53.
DOI: https://doi.org/10.1074/jbc.M410968200
Ehrhardt A, Chung WJ, Pyle LC, et al. Channel gating regulation by the cystic fibrosis transmembrane conductance regulator (CFTR) first cytosolic loop. J Biol Chem. 2016;291:1854-65.
DOI: https://doi.org/10.1074/jbc.M115.704809
Uppaluri L, England S, Scanlin T. Clinical evidence that V456A is a cystic fibrosis causing mutation in South Asians. J Cyst Fibros. 2012;11:312-5.
DOI: https://doi.org/10.1016/j.jcf.2012.02.001
Ziedalski TM, Kao PN, Henig NR, et al. Prospective analysis of cystic fibrosis transmembrane regulator mutations in adults with bronchiectasis or pulmonary nontuberculous mycobacterial infection. Chest. 2006;130:995-1002.
DOI: https://doi.org/10.1378/chest.130.4.995
McCormick J, Green MW, Mehta G, et al. Demographics of the UK cystic fibrosis population: implications for neonatal screening. Eur J Hum Genet. 2002;10:583-90.
DOI: https://doi.org/10.1038/sj.ejhg.5200850
Tsui LC, Dorfman R. The cystic fibrosis gene: a molecular genetic perspective. Cold Spring Harb Perspect Med. 2013;3:a009472.
DOI: https://doi.org/10.1101/cshperspect.a009472
Lukacs GL, Verkman AS. CFTR: folding, misfolding and correcting the DeltaF508 conformational defect. Trends Mol Med. 2012;18:81-91.
DOI: https://doi.org/10.1016/j.molmed.2011.10.003
Veit G, Avramescu RG, Chiang AN, et al. From CFTR biology toward combinatorial pharmacotherapy: expanded classification of cystic fibrosis mutations. Mol Biol Cell. 2016;27:424-33.
DOI: https://doi.org/10.1091/mbc.e14-04-0935
Lakshminarayan R, Phillips BP, Binnian IL, et al. Pre-emptive quality control of a misfolded membrane protein by ribosome-driven effects. Curr Biol. 2020;30:854-64 e5.
DOI: https://doi.org/10.1016/j.cub.2019.12.060
Ikuma M, Welsh MJ. Regulation of CFTR Cl- channel gating by ATP binding and hydrolysis. Proc Natl Acad Sci USA. 2000;97:8675-80.
DOI: https://doi.org/10.1073/pnas.140220597
How to Cite
Bakhat , K., Mateen, I., Saif, H., Anwar, K., Sarfraz, S., Javaid, S., ur Rehman , K., Arshad, A., & Mustafa, M. (2024). CFTR Exon 10 deleterious mutations in patients with congenital bilateral absence of vas deferens in a cohort of Pakistani patients. Archivio Italiano Di Urologia E Andrologia, 96(3). https://doi.org/10.4081/aiua.2024.12464
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