INTRODUCTION: Romani people have a high prevalence of kidney failure. This study examined a Romani cohort for pathogenic variants in the COL4A3, COL4A4, and COL4A5 genes that are affected in Alport syndrome (AS), a common cause of genetic kidney disease, characterized by hematuria, proteinuria, end-stage kidney failure, hearing loss, and eye anomalies. MATERIALS AND METHODS: The study included 57 Romani from different families with clinical features that suggested AS who underwent next-generation sequencing (NGS) of the COL4A3, COL4A4, and COL4A5 genes, and 83 family members. RESULTS: In total, 27 Romani (19%) had autosomal recessive AS caused by a homozygous pathogenic c.1598G>A, p.Gly533Asp variant in COL4A4 (n = 20) or a homozygous c.415G>C, p.Gly139Arg variant in COL4A3 (n = 7). For p.Gly533Asp, 12 (80%) had macroscopic hematuria, 12 (63%) developed end-stage kidney failure at a median age of 22 years, and 13 (67%) had hearing loss. For p.Gly139Arg, none had macroscopic hematuria (p = 0.023), three (50%) had end-stage kidney failure by a median age of 42 years (p = 0.653), and five (83%) had hearing loss (p = 0.367). The p.Gly533Asp variant was associated with a more severe phenotype than p.Gly139Arg, with an earlier age at end-stage kidney failure and more macroscopic hematuria. Microscopic hematuria was very common in heterozygotes with both p.Gly533Asp (91%) and p.Gly139Arg (92%). CONCLUSION: These two founder variants contribute to the high prevalence of kidney failure in Czech Romani. The estimated population frequency of autosomal recessive AS from these variants and consanguinity by descent is at least 1:11,000 in Czech Romani. This corresponds to a population frequency of autosomal dominant AS from these two variants alone of 1%. Romani with persistent hematuria should be offered genetic testing.

Department of Biology and Medical Genetics 2nd Faculty of Medicine Charles University and Motol University Hospital Prague Czechia

Department of Biomedical Sciences Faculty of Medicine University of Ostrava Ostrava Czechia

Department of Clinical and Molecular Pathology and Medical Genetics University Hospital Ostrava Ostrava Czechia

Department of Medical Biophysics Faculty of Medicine and Dentistry Palacký University Olomouc Olomouc Czechia

Department of Medical Genetics and Genomics University Hospital Brno Brno Czechia

Department of Medical Genetics Faculty of Medicine in Plzeň Charles University and University Hospital Plzeň Plzeň Czechia

Department of Medical Genetics Gennet s r o Liberec Czechia

Department of Medical Genetics Hospital České Budějovice a s České Budějovice Czechia

Department of Medical Genetics Krajská zdravotní a s Masaryk Hospital in Ústí nad Labem Ústí nad Labem Czechia

Department of Medical Genetics Thomayer University Hospital Prague Czechia

Department of Medical Genetics University Hospital Hradec Králové Hradec Králové Czechia

Department of Medicine The University of Melbourne Royal Melbourne Hospital Melbourne Australia

Department of Pediatrics 2nd Faculty of Medicine Charles University and Motol University Hospital Prague Czechia

Department of Pediatrics 3rd Faculty of Medicine Charles University and University Hospital Královské Vinohrady Prague Czechia

Department of Pediatrics Hospital Česká Lípa Česká Lípa Czechia

Institute of Clinical and Molecular Pathology Faculty of Medicine and Dentistry Palacký University Olomouc Olomouc Czechia

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacký University Olomouc Olomouc Czechia

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Jais J, Knebelmann B, Giatras I, Marchi M, Rizzoni G, Renieri A, et al. X-linked Alport syndrome: natural history in 195 families and genotype- phenotype correlations in males. J Am Soc Nephrol. (2000) 11:649–57. 10.1681/ASN.V114649 PubMed DOI

Hudson B, Tryggvason K, Sundaramoorthy M, Neilson E. Alport’s syndrome, Goodpasture’s syndrome, and type IV collagen. N Engl J Med. (2003) 348:2543–56. 10.1056/NEJMra022296 PubMed DOI

Kashtan C, Ding J, Garosi G, Heidet L, Massella L, Nakanishi K, et al. Alport syndrome: a unified classification of genetic disorders of collagen IV alpha345: a position paper of the Alport Syndrome Classification Working Group. Kidney Int. (2018) 93:1045–51. 10.1016/j.kint.2017.12.018 PubMed DOI

Papazachariou L, Papagregoriou G, Hadjipanagi D, Demosthenous P, Voskarides K, Koutsofti C, et al. Frequent COL4 mutations in familial microhematuria accompanied by later-onset Alport nephropathy due to focal segmental glomerulosclerosis. Clin Genet. (2017) 92:517–27. 10.1111/cge.13077 PubMed DOI

Savige J, Huang M, Dabrera M, Shukla K, Gibson J. Genotype-phenotype correlations for pathogenic COL4A3–COL4A5 variants in X-linked, autosomal recessive, and autosomal dominant Alport syndrome. Front Med. (2022) 9:865034. 10.3389/fmed.2022.865034 PubMed DOI PMC

Kluka V, Tischler V, Vereb J. [Alportś syndrome in Eastern Slovakia.] (Slovak). CS Pediat. (1986) 41:569–73. PubMed

Kolvek G, Podracka L, Rosenberger J, Stewart R, van Dijk J, Reijneveld S. Kidney diseases in Roma and non-Roma children from eastern Slovakia: are Roma children more at risk?. Int J Public Health. (2014) 59:1023–6. 10.1007/s00038-014-0609-z PubMed DOI

Zeman C, Depken D, Senchina D. Roma health issues: a review of the literature and discussion. Ethn Health. (2003) 8:223–49. 10.1080/1355785032000136434 PubMed DOI

Kalaydjieva L, Gresham D, Calafell F. Genetic studies of the Roma (gypsies): a review. BMC Med Genet. (2001) 2:5. 10.1186/1471-2350-2-5 PubMed DOI PMC

Sakhuja V, Sud K. End-stage renal disease in India and Pakistan: burden of disease and management issues. Kidney Int Suppl. (2003) 83:S115–8. 10.1046/j.1523-1755.63.s83.24.x PubMed DOI

Agarwal S, Srivastava R. Chronic kidney disease in India: challenges and solutions. Nephron Clin Pract. (2009) 111:c197–203. 10.1159/000199460 PubMed DOI

Lightstone L. Preventing renal disease: the ethnic challenge in the United Kingdom. Kidney Int Suppl. (2003) 83:S135–8. 10.1046/j.1523-1755.63.s83.29.x PubMed DOI

Barker D, Pruchno C, Jiang X, Atkin C, Stone E, Denison J, et al. A mutation causing Alport syndrome with tardive hearing loss is common in the western United States. Am J Hum Genet. (1996) 58:1157–65. PubMed PMC

Barker D, Denison J, Atkin C, Gregory M. Common ancestry of three Ashkenazi-American families with Alport syndrome and COL4A5 R1677Q. Hum Genet. (1997) 99:681–4. 10.1007/s004390050429 PubMed DOI

Webb B, Brandt T, Liu L, Jalas C, Liao J, Fedick A, et al. A founder mutation in COL4A3 causes autosomal recessive Alport syndrome in the Ashkenazi Jewish population. Clin Genet. (2014) 86:155–60. 10.1111/cge.12247 PubMed DOI

Zurowska A, Bielska O, Daca-Roszak P, Jankowski M, Szczepańska M, Roszkowska-Bjanid D, et al. Mild X-linked Alport syndrome due to the COL4A5 G624D variant originating in the Middle Ages is predominant in Central/East Europe and causes kidney failure in midlife. Kidney Int. (2021) 99:1451–8. 10.1016/j.kint.2020.10.040 PubMed DOI

Gibson J, Fieldhouse R, Chan M, Sadeghi-Alavijeh O, Burnett L, Izzi V, et al. Genomics England research consortiumprevalence estimates of predicted pathogenic COL4A3-COL4A5 variants in a population sequencing database and their implications for Alport syndrome. J Am Soc Nephrol. (2021) 32:2273–90. 10.1681/ASN.2020071065 PubMed DOI PMC

Stehlikova K, Skalova D, Zidkova J, Mrazova L, Vondravek P, Mazanec R, et al. Autosomal recessive limb-girdle muscular dystrophies in the Czech Republic. BMC Neurol. (2014) 14:154. 10.1186/s12883-014-0154-7 PubMed DOI PMC

Richards S, Aziz N, Bale S, Bick D, Das S, Gastier-Foster J, et al. ACMG laboratory quality assurance committee. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American college of medical genetics and genomics and the association for molecular pathology. Genet Med. (2015) 17:405–24. 10.1038/gim.2015.30 PubMed DOI PMC

Liu J, Wei X, Li A, Cui Y, Xia X, Qin W, et al. Novel mutations in COL4A3, COL4A4, and COL4A5 in Chinese patients with Alport Syndrome. PLoS One. (2017) 12:e0177685. 10.1371/journal.pone.0177685 PubMed DOI PMC

Storey H, Savige J, Sivakumar V, Abbs S, Flinter F. COL4A3/COL4A4 mutations and features in individuals with autosomal recessive Alport syndrome. J Am Soc Nephrol. (2013) 24:1945–54. 10.1681/ASN.2012100985 PubMed DOI PMC

Fokkema I, Taschner P, Schaafsma G, Celli J, Laros J, den Dunnen JT. LOVD v.2.0: the next generation in gene variant databases. Hum Mutat. (2011) 32:557–63. 10.1002/humu.21438 PubMed DOI

Wiszniewska J, Bi W, Shaw C, Stankiewicz P, Kang S, Pursley A, et al. Combined array CGH plus SNP genome analyses in a single assay for optimized clinical testing. Eur J Hum Genet. (2014) 22:79–87. 10.1038/ejhg.2013.77 PubMed DOI PMC

Romové, v Èeské republice. Czech radio [Romani in Czech Republic] (Czech) (1997–2023). Available online at: http://romove.radio.cz/cz/clanek/18884 (Accessed August 31, 2023).

Kolvek G, Rosicova K, Rosenberger J, Podracka L, Stewart R, Nagyova I, et al. End-stage renal disease among Roma and non-Roma: ROMA are at risk. Int J Public Health. (2012) 57:751–4. 10.1007/s00038-012-0365-x PubMed DOI

Savige J, Lipska-Zietkiewicz B, Watson E, Hertz J, Deltas C, Mari F, et al. Guidelines for genetic testing and management of Alport syndrome. Clin J Am Soc Nephrol. (2022) 17:143–54. 10.2215/CJN.04230321 PubMed DOI PMC

Dagher H, Wang Y, Fassett R, Savige J. Three novel COL4A4 mutations resulting in stop codons and their clinical effects in autosomal recessive Alport syndrome. Hum Mutat. (2002) 20:321–2. 10.1002/humu.9065 PubMed DOI

Oka M, Nozu K, Kaito H, Fu X, Nakanishi K, Hashimura Y, et al. Natural history of genetically proven autosomal recessive Alport syndrome. Pediatr Nephrol. (2014) 29:1535–44. 10.1007/s00467-014-2797-4 PubMed DOI

Lee J, Nozu K, Choi D, Kang H, Ha I, Cheong H. Features of autosomal recessive Alport syndrome: a systematic review. J Clin Med. (2019) 8:178. 10.3390/jcm8020178 PubMed DOI PMC

Kamiyoshi N, Nozu K, Fu X, Morisada N, Nozu Y, Ye M, et al. Genetic, clinical, and pathologic backgrounds of patients with autosomal dominant Alport syndrome. Clin J Am Soc Nephrol. (2016) 11:1441–9. 10.2215/CJN.01000116 PubMed DOI PMC

Matthaiou A, Poulli T, Deltas C. Prevalence of clinical, pathological and molecular features of glomerular basement membrane nephropathy caused by COL4A3 or COL4A4 mutations: a systematic review. Clin Kidney J. (2020) 13:1025–36. PubMed PMC

Furlano M, Martinez V, Pybus M, Arce Y, Crespi J, Venegas M, et al. Clinical and genetic features of autosomal dominant Alport syndrome: a cohort study. Am J Kidney Dis. (2021) 78:560–70. 10.1053/j.ajkd.2021.02.326 PubMed DOI

Gibson J, Huang M, Shenelli Croos Dabrera M, Shukla K, Rothe H, Hilbert P, et al. Genotype-phenotype correlations for COL4A3-COL4A5 variants that result in Gly substitutions in Alport syndrome. Sci Rep. (2022) 12:2722. 10.1038/s41598-022-06525-9 PubMed DOI PMC

Bouwer S, Angelicheva D, Chandler D, Seeman P, Tournev I, Kalaydjieva L. Carrier rates of the ancestral Indian W24X mutation in GJB2 in the general gypsy population and individual subisolates. Genet Test. (2007) 11:455–8. 10.1089/gte.2007.0048 PubMed DOI

Safka Brozkova D, Lastuvkova J, Stepankova H, Krutova M, Trkova M, Myska P, et al. DFNB49 is an important cause of non-syndromic deafness in Czech Roma patients but not in the general Czech population. Clin Genet. (2012) 82:579–82. 10.1111/j.1399-0004.2011.01817.x PubMed DOI

Safka Brozkova D, Varga L, Uhrova Meszarosova A, Slobodova Z, Skopkova M, Soltysova A, et al. Variant c.2158-2A>G in MANBA is an important and frequent cause of hereditary hearing loss and beta-mannosidosis among the Czech and Slovak Roma population- evidence for a new ethnic-specific variant. Orphanet J Rare Dis. (2020) 15:222. 10.1186/s13023-020-01508-3 PubMed DOI PMC