SUMMARY. The elucidation of molecular genetic mechanisms in the development of rare inherited diseases is crucial in understanding the processes of pathogenesis, improving the diagnosis and treatment of patients. The aim of our study was to determine the nature of genetic defects in a patient who showed signs of sexual dysfunction in combination with neuromuscular pathology. The study included: clinical and genealogical examination, ultrasonographic examination, electroneuromyography, study of biochemical parameters in serum: hormones and enzymes, molecular cytogenetic study using the FISH method, whole exome sequencing of the patient's DNA analysis, bioinformatiс analysis. The patient showed signs of primary (hypergonadotropic) hypogonadism and polyneuropathic changes in the lower extremities (axonal-demyelinating). It was found that these clinical characteristics of the patient's phenotype do not agree with any description of the clinical phenotype registered in the OMIM International Database. According to the results of molecular cytogenetic study in the patient's karyotype, an unbalanced translocation of the Yp11.3 region containing the SRY gene to the short arm region of the X chromosome Xp22.33 with the formation of a derivative X chromosome was detected. On another side, the results of whole exome sequencing of the patient's genomic DNA revealed the presence of a homozygous mutation chr5: 131165096 C> G (c.110G> C, p.Arg37Pro) in the HINT1 gene. Thus, it can be concluded that the phenotype with signs of disorders of sex differentiation in combination with axonal neuropathy with myotonia is due to two independent pathogenetic factors - unbalanced translocation of the SRY gene sequence on the X chromosome, resulting in paternal spermatogenesis and inheritance of the HINT1 gene homozygous missence mutation (c.110G>C, p.Arg37Pro). These results support the effectiveness of a combination of molecular cytogenetic research methods and the results of high-performance WES to elucidate the molecular mechanisms of pathogenesis in patients with rare clinical phenotypes.
Keywords: disorder of sex differentiation, neuromuscular disorder, chromosomal unbalanced translocation, missense mutation, SRY, HINT1, Х chromosome
Full text and supplemented materials
References
Bieganowski, P., Garrison, P.N., Hodawadekar, S.C., Faye, G., Barnes, L.D., and Brenner, Ch., Show footnotes adenosine monophosphoramidase activity of Hint and Hnt1 supports function of Kin28, Ccl1, and Tfb3, Mech. Signal Transduction, 2002, vol. 277, no. 13, pp. 10852–10860.
Boaretto, F., Cacciavillani, M., Mostacciuolo, M.L., Spalletta, A., Piscosquito, G., Pareyson, D., Vazza, G., and Briani, C., Novel loss-of-function pathogenic variant of the HINT1 gene in a patient with distal motor axonal neuropathy without neuromyotonia, Muscle Nerve, 2015, vol. 52, no. 4, pp. 688–689. https://doi.org/10.1002/mus.24720
Braga, B., Gomes, N., Nishi, M., Freire, B., et al., Variants in 46, XY DSDRelated genes in syndromic and non-syndromic small for gestational age children with hypospadias, Sex. Dev., 2022, vol. 16, no. 1, pp. 27–33. https://doi.org/10.1159/000518091
Brenner, Ch., Hint, Fhit, and GalT: function, structure, evolution, and mechanism of three branches of the histidine triad superfamily of nucleotide hydrolases and transferases, Biochemistry, 2002, vol. 41, no. 29, pp. 9003–9014. https://doi.org/10.1021/bi025942q
Chou, T.F. and Wagner, C.R., Lysyl-tRNA synthetase-generated lysyl-adenylate is a substrate for histidine triad nucleotide binding proteins, J. Biol. Chem., 2007, vol. 282, no. 7, pp. 4719–4727. https://doi.org/10.1074/jbc.M610530200
Chou, T.F., Tikh, I.B., Horta, B.A., et al., Engineered monomeric human histidine triad nucleotide-binding protein 1 hydrolyzes fluorogenic acyl-adenylate and lysyl-tRNA synthetase-generated lysyl-adenylate, J. Biol. Chem., 2007, vol. 282, no. 20, pp. 15137–15147.
Eggers, S., Ohnesorg, T., and Sinclair, A., Genetic regulation of mammalian gonad development, Nat. Rev. Endocrinol., 2014, vol. 10, pp. 673–683.
Ergun-Longmire, B., Vinci, G., Alonso, L., Matthew, S., Tansil, S., Lin-Su, K., McElreavey, K., and New, M.I., Clinical, hormonal and cytogenetic evaluation of 46, XX males and review of the literature, J. Pediatr. Endocrinol. Metab., 2005, vol. 18, no. 8, pp. 739–748. https://doi.org/10.1515/jpem.2005.18.8.739
Estermann, M.A. and Smith, C.A., Applying single-cell analysis to gonadogenesis and DSDs (Disorders/Differences of Sex Development), Int. J. Mol. Sci., 2020, vol. 21, no. 18, art. ID 6614.
Ilaslan, E., Markosyan, R., Sproll, P., Stevenson, B.J., Sajek, M., Sajek, M.P., Hayrapetyan, H., Sarkisian, T., Livshits, L., Nef, S., Jaruzelska, J., and Kusz-Zamelczyk, K., The FKBP4 gene, encoding a regulator of the androgen receptor signaling pathway, is a novel candidate gene for androgen insensitivity syndrome, Int. J. Mol. Sci., 2020, vol. 21, no. 21, art. ID 8403. https://doi.org/10.3390/ijms21218403
Lamothe, S., Bernard, V., and Christin-Maitre, S., Gonad differentiation toward ovary, Ann. d’Endocrinol., 2020, vol. 81, nos. 2–3, pp. 83–88.
Laššuthová, P., Brožková, D.Š., Krůtová, M., Neupauerová, J., Haberlová, J., Mazanec, R., Dvořáčková, N., Goldenberg, Z., and Seeman, P., Pathogenic variants in HINT1 are one of the most frequent causes of hereditary neuropathy among Czech patients and neuromyotonia is rather an underdiagnosed symptom, Neurogenetics, 2015, vol. 16, no. 1, pp. 43–54. https://doi.org/10.1007/s10048-014-0427-8
Madeiro, B. de A.C.S., Peeters, K., Santos de Lima, E.L., Amor-Barris, S., Els, De Vriendt, Jordanova, A., Cartaxo Muniz, M.T., da Cunha Correia, C., HINT1 founder mutation causing axonal neuropathy with neuromyotonia in South America: A case report, Mol. Genet. Genomic Med., 2021, vol. 9, no. 10, art. ID e1783.https://doi.org/10.1002/mgg3.1783
Majzoub, A., Arafa, M., Starks, Ch., Elbardisi, H., Said, S.Al., and Sabanegh, E., 46,XX karyotype during male fertility evaluation; case series and literature review, Asian J. Androl., 2017, vol. 19, no. 2, pp. 168–172.
Martínez de LaPiscina, I., Mahmoud, R.A., Sauter, K.-S., Esteva, I., Alonso, M., Costa, I., Rial-Rodriguez, J.M., Rodríguez-Estévez, A., Vela, A., Castano, L., and Flück, C.E., Variants of STAR, AMH and ZFPM2/ FOG2 may contribute towards the broad phenotype observed in 46,XY DSD patients with heterozygous variants of NR5A1, Int. J Mol Sci., 2020, vol. 21, no. 22, art. ID 8554.
Nøstvik, M., Kateta, S.M., Schönewolf-Greulich, B., Barth, A.A.M., Boschann, F., Doummar, D., Haack, T.B., Keren, B., Livshits, L.A., Mei, D., Park, J., Pisano, T., Prouteau, C., Umair, M., Waqas, A., Ziegler, A., Guerrini, R., Moller, R.S., and Tümer, Z., Clinical and molecular delineation of PUS3-associated neurodevelopmental disorders, Clin. Genet., 2021, vol. 100, no. 5, pp. 628–633. https://doi.org/10.1111/cge.14051
Queralt, R., Madrigal, I., Vallecillos, M.A., Morales, C., Ballescá, J.-L., Oliva, R., Soler, A., Sánchez, A., and Margarit, E., Atypical XX male with the SRY gene located at the long arm of chromosome 1 and a 1qter microdeletion, Am. J. Med. Genet., 2008, vol. 146A, no. 10, pp. 1335–1340.
Rayevsky, A., Sirokha, D., Samofalova, D., Lozhko, D., Gorodna, O., Prokopenko, I., Livshits, L., Functional effects in silico prediction for androgen receptor ligand-binding domain novel I836S mutation, Life, 2021, vol. 11, no. 7, art. ID 659. https://doi.org/10.3390/life11070659
Shchagina, O.A., Milovidova, T.B., Murtazina, A.F., Rudenskaya, G.E., Nikitin, S.S., Dadali, E.L., and Polyakov, A.V., HINT1 gene pathogenic variants: the most common cause of recessive hereditary motor and sensory neuropathies in Russian patient, Mol. Biol. Rep., 2019, vol. 47, pp. 1331–1337. https://doi.org/10.1007/s11033-019-05238-z
Shu-Chin, Chien, Yueh-Chun, Li, Ming, Ho, Pei-Ching, Hsu, Ru-Hsiou, Teng, Wei-De, Lin, Fuu-Jen, Tsai, and Chyi-Chyang, Lin, Rare rearrangements: A “jumping satellite” in one family and autosomal location of the SRY gene in an XX male, Am. J. Med. Genet., 2009, vol. 149A, no. 12, art. ID 2775–2781.
Sirokha, D., Gorodna, O., Vitrenko, Y., Zelinska, N., Ploski, R., Nef, S., Jaruzelska, J., Kusz-Zamelczyk, K., and Livshits, L., A novel WT1 mutation identified in a 46,XX testicular/ovotesticular DSD patient results in the retention of intron 9, Biology, 2021, vol. 10, no. 12, art. ID 1248. https://doi.org/10.3390/biology10121248
Soloviov, O.O., Livshits, G.B., Podlesnaya, S.S., and Livshits, L.A., Implementation of the quantitative Real-Time PCR for the molecular-genetic diagnostics of spinal muscular atrophy, Biopolym. Cell, 2010, vol. 26, no. 1, pp. 51–55.
Weiske, J. and Huber, O., The histidine triad protein Hint1 interacts with Pontin and Reptin and inhibits TCF–β-catenin-mediated transcription, J. Cell Sci., 2005, vol. 118, no. 14, pp. 3117–3129. https://doi.org/10.1242/jcs.02437
Zhao, H., Race, V., Matthijs, G., De Jonghe, P., Robberecht, W., Lambrechts, D., and Van Damme, P., Exome sequencing reveals HINT1 pathogenic variants as a cause of distal hereditary motor neuropathy, Eur. J. Hum. Genet., 2014, vol. 22, no. 6, pp. 847–850. https://doi.org/10.1038/ejhg.2013.231
Zimoń, M., Baets, J., Almeida-Souza, L., et al., Loss-of-function mutations in HINT1 cause axonal neuropathy with neuromyotonia, Nat. Genet., 2012, vol. 44, no. 10, pp. 1080–1083. https://doi.org/10.1038/ng.2406