Liver as a source for thymidine phosphorylase replacement in mitochondrial neurogastrointestinal encephalomyopathy

PLoS One. 2014 May 6;9(5):e96692. doi: 10.1371/journal.pone.0096692. eCollection 2014.

Abstract

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare autosomal recessive mitochondrial disease associated with mutations in the nuclear TYMP gene. As a result, the thymidine phosphorylase (TP) enzyme activity is markedly reduced leading to toxic accumulation of thymidine and therefore altered mitochondrial DNA. MNGIE is characterized by severe gastrointestinal dysmotility, neurological impairment, reduced life expectancy and poor quality of life. There are limited therapeutic options for MNGIE. In the attempt to restore TP activity, allogenic hematopoietic stem cell transplantation has been used as cellular source of TP. The results of this approach on ∼ 20 MNGIE patients showed gastrointestinal and neurological improvement, although the 5-year mortality rate is about 70%. In this study we tested whether the liver may serve as an alternative source of TP. We investigated 11 patients (7M; 35-55 years) who underwent hepatic resection for focal disorders. Margins of normal liver tissue were processed to identify, quantify and localize the TP protein by Western Blot, ELISA, and immunohistochemistry, and to evaluate TYMP mRNA expression by qPCR. Western Blot identified TP in liver with a TP/GAPDH ratio of 0.9 ± 0.5. ELISA estimated TP content as 0.5 ± 0.07 ng/μg of total protein. TP was identified in both nuclei and cytoplasm of hepatocytes and sinusoidal lining cells. Finally, TYMP mRNA was expressed in the liver. Overall, our study demonstrates that the liver is an important source of TP. Orthotopic liver transplantation may be considered as a therapeutic alternative for MNGIE patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Blotting, Western
  • Duodenum / enzymology
  • Duodenum / metabolism
  • Duodenum / pathology
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Immunohistochemistry
  • Liver / enzymology
  • Liver / metabolism
  • Liver / pathology
  • Male
  • Middle Aged
  • Mitochondrial Encephalomyopathies / genetics
  • Mitochondrial Encephalomyopathies / pathology*
  • Muscle, Skeletal / enzymology
  • Muscle, Skeletal / metabolism
  • Muscle, Skeletal / pathology
  • RNA, Messenger / metabolism
  • Thymidine Phosphorylase / genetics
  • Thymidine Phosphorylase / metabolism*

Substances

  • RNA, Messenger
  • Thymidine Phosphorylase

Grants and funding

This work was partly supported by grants from the Italian Ministry of Health (Ricerca Finalizzata 2009 RF2009) and funds from University of Bologna (R. De G.). R. De G., V. T. and E. B. were the recipients of grants from “Fondazione Del Monte di Bologna e Ravenna”, Bologna, Italy. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.