Skip to main content
SpringerLink
Log in

Variable X-chromosome inactivation and enlargement of pericentral glutamine synthetase zones in the liver of heterozygous females with OTC deficiency

  • Original Article
  • Published:
Virchows Archiv Aims and scope Submit manuscript

Abstract

Ornithine transcarbamylase (OTC) deficiency is an X-linked disorder that causes recurrent and life-threatening episodes of hyperammonemia. The clinical picture in heterozygous females is highly diverse and derives from the genotype and the degree of inactivation of the mutated X chromosome in hepatocytes. Here, we describe molecular genetic, biochemical, and histopathological findings in the livers explanted from two female patients with late-onset OTC deficiency. Analysis of X-inactivation ratios by DNA methylation-based assays showed remarkable intra-organ variation ranging from 46:54 to 82:18 (average 70:30, n = 37), in favor of the active X chromosome carrying the mutation c.583G>C (p.G195R), in the first patient and from 75:25 to 90:10 (average 82:18, n = 20) in favor of the active X chromosome carrying the splicing mutation c.663+1G>A in the second patient. The X-inactivation ratios in liver samples correlated highly with the proportions of OTC-positive hepatocytes calculated from high-resolution image analyses of the immunohistochemically detected OTC in frozen sections that was performed on total area > 5 cm2. X-inactivation ratios in blood in both female patients corresponded to the lower limit of the liver values. Our data indicate that the proportion of about 20–30% of hepatocytes expressing the functional OTC protein is not sufficient to maintain metabolic stability. X-inactivation ratios assessed in liver biopsies taken from heterozygous females with X-linked disorders should not be considered representative of the whole liver.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Yu W, Lin Y, Yao J, Huang W, Lei Q, Xiong Y, Zhao S, Guan KL (2009) Lysine 88 acetylation negatively regulates ornithine carbamoyltransferase activity in response to nutrient signals. J Biol Chem 274:13669–13675. https://doi.org/10.1074/jbc.M901921200

    Article  CAS  Google Scholar 

  2. Hallows WC, Yu W, Smith BC, Devires MK, Ellinger JJ, Someya S, Shortreed MR, Prolla T, Markley JL, Smith LM, Zhao S, Guan KL, Denu JM (2011) Sirt3 promotes the urea cycle and fatty acid oxidation during dietary restriction. Mol Cell 41:139–149. https://doi.org/10.1016/j.molcel.2011.01.002

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  3. Nakagawa T, Lomb DJ, Haigis MC, Guarente L (2009) SIRT5 deacetylates carbamoyl phosphate synthetase 1 and regulates the urea cycle. Cell 137:560–570. https://doi.org/10.1016/j.cell.2009.02.026

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Gebhardt R, Matz-Soja M (2014) Liver zonation: novel aspects of its regulation and its impact on homeostasis. World J Gastroenterol 20:8491–8504. https://doi.org/10.3748/wjg.v20.i26.8491

    Article  PubMed  PubMed Central  Google Scholar 

  5. Bahar Halpern K, Shenhav R, Matcovitch-Natan O et al (2017) Single-cell spatial reconstruction reveals global division of labour in the mammalian liver. Nature 542:352–356. https://doi.org/10.1038/nature21065

    Article  CAS  Google Scholar 

  6. Torre C, Perret C, Colnot S (2010) Molecular determinants of liver zonation. Prog Mol Biol Transl Sci 97:127–150. https://doi.org/10.1016/B978-0-12-385233-5.00005-2

    Article  PubMed  CAS  Google Scholar 

  7. Cavicchi C, Donati M, Parini R et al (2014) Sudden unexpected fatal encephalopathy in adults with OTC gene mutations—clues for early diagnosis and timely treatment. Orphanet J Rare Dis 9:105. https://doi.org/10.1186/s13023-014-0105-9

    Article  PubMed  PubMed Central  Google Scholar 

  8. Yorifuji T, Muroi J, Uematsu A, Tanaka K, Kiwaki K, Endo F, Matsuda I, Nagasaka H, Furusho K (1998) X-inactivation pattern in the liver of a manifesting female with ornithine transcarbamylase (OTC) deficiency. Clin Genet 54:349–353

    Article  PubMed  CAS  Google Scholar 

  9. Lichter-Konecki U, Caldovic L, Morizono H, et al (2016) Ornithine transcarbamylase deficiency. In: Pagon RA, Adam MP, Ardinger HH, et al., editors. GeneReviews® [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2017. Avalaible from: https://www.ncbi.nlm.nih.gov/books/NBK154378/

  10. Storkanova G, Vlaskova H, Chuzhanova N, Zeman J, Stranecky V, Majer F, Peskova K, Luksan O, Jirsa M, Hrebicek M, Dvorakova L (2013) Ornithine carbamoyltransferase deficiency: molecular characterization of 29 families. Clin Genet 84:552–559. https://doi.org/10.1111/cge.12085

    Article  PubMed  CAS  Google Scholar 

  11. Musalkova D, Minks J, Storkanova G, Dvorakova L, Hrebicek M (2015) Identification of novel informative loci for DNA-based X-inactivation analysis. Blood Cells Mol Dis 54:210–216. https://doi.org/10.1016/j.bcmd.2014.10.001

    Article  PubMed  CAS  Google Scholar 

  12. Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R, Salzberg SL (2013) TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol 14:R36. https://doi.org/10.1186/gb-2013-14-4-r36

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  13. Li H (2011) A statistical framework for SNP calling, mutation discovery, association mapping and population genetical parameter estimation from sequencing data. Bioinformatics 27:2987–2993. https://doi.org/10.1093/bioinformatics/btr509

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  14. Tuchman M, Tsai MY, Holzkneccht RA, Brusilow SW (1989) Carbamyl phosphate synthetase and ornithine transcarbamylase activities in enzyme-deficient human liver measured by radiochromatography and correlated with outcome. Pediatr Res 26:77–82. https://doi.org/10.1203/00006450-198907000-00021

    Article  PubMed  CAS  Google Scholar 

  15. Hers HG (1964) Glycogen storage disease. Adv Metab Disord 13:1–44

    PubMed  CAS  Google Scholar 

  16. Allen RC, Zoghbi HY, Moseley AB, Rosenblatt HM, Belmont JW (1992) Methylation of HpaII and HhaI sites near the polymorphic CAG repeat in the human androgen-receptor gene correlates with X chromosome inactivation. Am J Hum Genet 51:1229–1239

    PubMed  PubMed Central  CAS  Google Scholar 

  17. Kogo T, Satoh Y, Kanazawa M, Yamamoto S, Takayanagi M, Ohtake A, Mori M, Niimi H (1998) Expression analysis of two mutant human ornithine transcarbamylases in COS-7 cells. J Hum Genet 43:54–58. https://doi.org/10.1007/s100380050037

    Article  PubMed  CAS  Google Scholar 

  18. Choi JH, Lee BH, Kim JH, Kim GH, Kim YM, Cho J, Cheon CK, Ko JM, Lee JH, Yoo HW (2015) Clinical outcomes and the mutation spectrum of the OTC gene in patients with ornithine transcarbamylase deficiency. J Hum Genet 60:501–507. https://doi.org/10.1038/jhg.2015.54

    Article  PubMed  CAS  Google Scholar 

  19. Kim GH, Choi JH, Lee HH, Park S, Kim SS, Yoo HW (2006) Identification of novel mutations in the human ornithine transcarbamylase (OTC) gene of Korean patients with OTC deficiency and transient expression of the mutant proteins in vitro. Hum Mutat 27:1159. https://doi.org/10.1002/humu.9465

    Article  PubMed  Google Scholar 

  20. Tuchman M, Plante RJ, McCann MT, Qureshi AA (1994) Seven new mutations in the human ornithine transcarbamylase gene. Hum Mutat 4:57–60. https://doi.org/10.1002/humu.1380040109

    Article  PubMed  CAS  Google Scholar 

  21. Rahayatri T, Uchida H, Sasaki K et al (2017) Hyperammonemia in ornithine transcarbamylase-deficient recipients following living donor liver transplantation from heterozygous carrier donors. Pediatr Transplant 21:e12848. https://doi.org/10.1111/petr.12848

    Article  CAS  Google Scholar 

  22. De Hoon B, Monkhorst K, Riegman P, Laven JS, Gribnau J (2015) Buccal swab as a reliable predictor for X inactivation ratio in inaccessible tissues. J Med Genet 52:784–790. https://doi.org/10.1136/jmedgenet-2015-103194

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Wakiya T, Sanada Y, Urahashi T, Ihara Y, Yamada N, Okada N, Egami S, Sakamoto K, Murayama K, Hakamada K, Yasuda Y, Mizuta K (2012) Living donor liver transplantation from an asymptomatic mother who was a carrier for ornithine transcarbamylase deficiency. Pediatr Transplant 16:E196–E200. https://doi.org/10.1111/j.1399-3046.2012.01716.x

    Article  PubMed  CAS  Google Scholar 

  24. Wakiya T, Sanada Y, Urahashi T, Ihara Y, Yamada N, Okada N, Ushijima K, Otomo S, Sakamoto K, Murayama K, Takayanagi M, Hakamada K, Yasuda Y, Mizuta K (2012) Impact of enzyme activity assay on indication in liver transplantation for ornithine transcarbamylase deficiency. Mol Genet Metab 105:404–407. https://doi.org/10.1016/j.ymgme.2011.12.019

    Article  PubMed  CAS  Google Scholar 

  25. Badizadegan K, Perez-Atayde AR (1997) Focal glycogenosis of the liver in disorders of ureagenesis: its occurrence and diagnostic significance. Hepatology 26:365–373. https://doi.org/10.1002/hep.510260217

    Article  PubMed  CAS  Google Scholar 

  26. Yaplito-Lee J, Chow CW, Boneh A (2013) Histopathological findings in livers of patients with urea cycle disorders. Mol Genet Metab 108:161–165. https://doi.org/10.1016/j.ymgme.2013.01.006

    Article  PubMed  CAS  Google Scholar 

  27. Wu H, Luo J, Yu H, Rattner A, Mo A, Wang Y, Smallwood PM, Erlanger B, Wheelan SJ, Nathans J (2014) Cellular resolution maps of X chromosome inactivation: implications for neural development, function, and disease. Neuron 81:103–119. https://doi.org/10.1016/j.neuron.2013.10.051

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Hedberg Oldfors C, Máthé G, Thomson K, Tulinius M, Karason K, Östman-Smith I, Oldfors A (2015) Early onset cardiomyopathy in females with Danon disease. Neuromuscul Disord 25:493–501. https://doi.org/10.1016/j.nmd.2015.03.005

    Article  PubMed  Google Scholar 

  29. Miles L, Heubi JE, Bove KE (2005) Hepatocyte glycogen accumulation in patients undergoing dietary management of urea cycle defects mimics storage disease. J Pediatr Gastroenterol Nutr 40:471–476

    Article  PubMed  CAS  Google Scholar 

  30. Carpentieri D, Barnhart MF, Aleck K, Miloh T, deMello D (2015) Lysinuric protein intolerance in a family of Mexican ancestry with a novel SLC7A7 gene deletion. Case report and review of the literature. Mol Genet Metab Rep 2:47–50. https://doi.org/10.1016/j.ymgmr.2014.12.005

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Burrage L, Madan S, Li X, Jiang M, Cela R, Finegold M, Nagamani S, Bali D, Lee B. Argininosuccinate lyase deficiency: a hepatic glycogen storage disorder? Poster presented at ASHG 2016 Meeting. 2016 October 18–22; Vancouver, Canada. Retrieved from http://www.ashg.org/2016meeting/listing/PosterSessions.shtml, 27th March 2017

  32. Shiojiri N, Imai H, Goto S, Ohta T, Ogawa K, Mori M (1997) Mosaic pattern of ornithine transcarbamylase expression in spfash mouse liver. Am J Pathol 151:413–421

    PubMed  PubMed Central  CAS  Google Scholar 

  33. Matsuda I, Tanase S (1997) The ornithine transcarbamylase (OTC) gene: mutations in 50 Japanese families with OTC deficiency. Am J Med Genet 71:378–383

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The authors would like to thank the patients and their families. We acknowledge the help and advice provided by Marketa Novakova, Michaela Hnizdova Bouckova, Michaela Fialova, Gabriela Storkanova, Ondrej Luksan, Karolina Peskova, and particularly, by Lenka Piherova and Jakub Sikora.

Funding

This study was supported by the projects GA UK No. 42314 and No. 580716 from the Grant Agency of Charles University in Prague, and also by projects SVV 260367, UNCE 204011/2012, Progres Q26, and MH CZ-DRO VFN 64165.

Author information

Authors and Affiliations

  1. Research Unit for Rare Diseases, Department of Pediatrics and Adolescent Medicine, First Faculty of Medicine, Charles University and General University Hospital in Prague, Ke Karlovu 2, 128 08, Prague 2, Czech Republic

    Dita Musalkova, Martin Reboun, Jitka Sokolova, Jakub Krijt, Jitka Honzikova, Jiri Gurka, Lenka Dvorakova & Martin Hrebicek

  2. Department of Clinical and Transplant Pathology, Institute for Clinical and Experimental Medicine (IKEM), Prague, Czech Republic

    Eva Sticova

  3. Institute of Histology and Embryology, First Faculty of Medicine, Charles University, Prague, Czech Republic

    Jiri Gurka

  4. Laboratory of Experimental Hepatology, Institute for Clinical and Experimental Medicine (IKEM), Prague, Czech Republic

    Magdalena Neroldova & Milan Jirsa

  5. Department of Pediatrics and Adolescent Medicine, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic

    Tomas Honzik & Jiri Zeman

Authors
  1. Dita Musalkova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Eva Sticova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Martin Reboun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jitka Sokolova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jakub Krijt
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jitka Honzikova
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jiri Gurka
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Magdalena Neroldova
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Tomas Honzik
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Jiri Zeman
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Milan Jirsa
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Lenka Dvorakova
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Martin Hrebicek
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Dita Musalkova or Martin Hrebicek.

Ethics declarations

The study was approved by the local ethics committee and conducted in agreement with institutional guidelines. Written informed consent was obtained from all adult study participants. On behalf of the patient written informed consent was obtained from her parents.

Conflict of interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

ESM 1

(PDF 168 kb)

ESM 2

(PDF 704 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Musalkova, D., Sticova, E., Reboun, M. et al. Variable X-chromosome inactivation and enlargement of pericentral glutamine synthetase zones in the liver of heterozygous females with OTC deficiency. Virchows Arch 472, 1029–1039 (2018). https://doi.org/10.1007/s00428-018-2345-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00428-018-2345-x

Keywords

Search

Navigation