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Lack of global epigenetic methylation defects in CBS deficient mice

  • Original Article
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Journal of Inherited Metabolic Disease

Abstract

Cystathionine β-synthase (CBS) deficiency is a recessive inborn error of metabolism in which patients have extremely elevated plasma total homocysteine and have clinical manifestations in the vascular, visual, skeletal, and nervous systems. Homocysteine is an intermediary metabolite produced from the hydrolysis of S-adenosylhomocysteine (SAH), which is a by-product of methylation reactions involving the methyl-donor S-adenosylmethionine (SAM). Here, we have measured SAM, SAH, DNA and histone methylation status in an inducible mouse model of CBS deficiency to test the hypothesis that homocysteine-related phenotypes are caused by inhibition of methylation due to elevated SAH and reduced SAM/SAH ratio. We found that mice lacking CBS have elevated cellular SAH and reduced SAM/SAH ratios in both liver and kidney, but this was not associated with alterations in the level of 5-methylcytosine or various histone modifications. Using methylated DNA immunoprecipitation in combination with microarray, we found that of the 241 most differentially methylated promoter probes, 89 % were actually hypermethylated in CBS deficient mice. In addition, we did not find that changes in DNA methylation correlated well with changes in RNA expression in the livers of induced and uninduced CBS mice. Our data indicates that reduction in the SAM/SAH ratio, due to loss of CBS activity, does not result in overall hypomethylation of either DNA or histones.

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Acknowledgments

This work was funded in part by the following grants from the National Institutes of Health: NIH (CA06927 and R01GM098772), and an appropriation from the Commonwealth of Pennsylvania. We also thank the Genomics and Laboratory Animal Facilities of Fox Chase Cancer Center for their assistance. We also acknowledge secretarial support by Ms. Kathy Ireton.

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Authors and Affiliations

  1. Cancer Biology Program, Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA, 19111, USA

    Hyung-Ok Lee, Liqun Wang, Yin-Ming Kuo, Sapna Gupta, Yue-sheng Li, Andrew J. Andrews & Warren D. Kruger

  2. Biostatisitics Program, Fox Chase Cancer Center, Philadelphia, PA, USA

    Michael J. Slifker

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  1. Hyung-Ok Lee
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  2. Liqun Wang
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  3. Yin-Ming Kuo
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  8. Warren D. Kruger
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Correspondence to Warren D. Kruger.

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Competing interest

Hyung-Ok Lee, Liqun Wang, Yin-Ming Kuo, Sapna Gupta, Michael J. Slifker, Yue-sheng Li, Andrew J. Andrews, and Warren D. Kruger all declare that that they have no conflict of interest.

All institutional and national guidelines for the care and use of laboratory animals were followed.

No human subjects were used in these studies.

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Responsible editor: Viktor Kozich

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Supplemental Fig. 1

The correlation between tHcy and tissue SAM, SAH, and SAM/SAH ratios in the liver and kidney. P values show slope significant in linear regression analysis (PPTX 3926 kb)

Supplemental Fig. 2

MeDIP-Chip analysis. (a) Histogram showing the LogFC of all the probes. The solid curve line shows the expected normal distribution centered around the mean. (b) Same as A, but log scale to better show the enrichment of probes with –LogFC values. (c) Scatter plot showing the relationship between the LogFC of the 241 probes with P < 0.01 and the corresponding LogFC of their RNAs. Regression line with 95 % confidence interval is shown. (d) Scatter plot of LogFC of the RNA 20 most differentially regulated genes and the LogFC of all the methylated probes within those genes (PPTX 201 kb)

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Lee, HO., Wang, L., Kuo, YM. et al. Lack of global epigenetic methylation defects in CBS deficient mice. J Inherit Metab Dis 40, 113–120 (2017). https://doi.org/10.1007/s10545-016-9958-5

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  • DOI: https://doi.org/10.1007/s10545-016-9958-5

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