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Functional Restoration of Amyotrophic Lateral Sclerosis Patient-Derived Mesenchymal Stromal Cells Through Inhibition of DNA Methyltransferase

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Abstract

Alteration of DNA methylation is highly associated with aging and neurodegenerative disorders, such as amyotrophic lateral sclerosis (ALS). Remedying these aberrant methylation patterns may serve to improve these diseases. Previously, we reported that human bone marrow mesenchymal stromal cells isolated from ALS patients (ALS-MSCs) have functionally decreased stem cell potency, and excessively express DNA methyltransferases (DNMTs). In this study, we examined the correlation between excessive DNMT expression and functional decline in ALS-MSCs. The DNMT inhibitor RG108 was used for this. RG108-treated ALS-MSCs exhibit increased expression of the anti-senescence genes TERT, VEGF, and ANG, and decreased expression of the senescence-related genes ATM and p21. The activity of SA-β-galactosidase and the expression of senescence proteins p53 and p16 were reduced in RG108-treated ALS-MSCs. The abilities of cell migration and protection against oxidative damage were improved in the treated ALS-MSCs. In neuronal differentiation experiments, the treated MSCs more effectively differentiated into neuron-like cells. These results suggest that ALS-MSC function can be restored by inhibiting excessively expressed DNMTs, an approach that may ultimately provide better efficacy in stem cell therapy.

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Abbreviations

ALS:

Amyotrophic lateral sclerosis

ANG:

Angiogenin

ATM:

Ataxia telangiectasia mutated

BHA:

Butyalated hydroxyanisole

BM-MSCs:

Bone marrow mesenchymal stromal cells

DMSO:

Dimethyl sulfoxide

DNMTs:

DNA methyltransferases

FBS:

Fetal bovine serum

MMPs:

Metalloproteinase

MSCs:

Mesenchymal stromal cells

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PBS:

Phosphate-buffered saline

PCR:

Polymerase chain reaction

RIPA:

Radioimmunoprecipitation assay

SA-β-gal staining:

Senescence-associated beta-galactosidase staining

TERT:

Telomerase

VEGF:

Vascular endothelial growth factor

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Acknowledgments

This work was supported by a grant from the Korea Healthcare Technology R&D Project, Ministry for Health, Welfare & Family Affairs, Republic of Korea (A120203), a grant from the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2010-0010431).

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Authors and Affiliations

  1. Department of Biology, College of Natural Science, Chosun University, 309 Pilmun-daero, Dong-gu, Gwangju, 501-759, Korea

    Youn Seo Oh & Goang-Won Cho

  2. Department of Life Science, BK21-Plus Research Team for Bioactive Control Technology, Chosun University, Gwangju, 501-759, Korea

    Youn Seo Oh & Goang-Won Cho

  3. Department of Neurology, Hanyang University College of Medicine, Seoul, Korea

    Seung Hyun Kim

  4. Cell Therapy Center, Hanyang University Hospital, Seoul, Korea

    Seung Hyun Kim

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  1. Youn Seo Oh
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Correspondence to Goang-Won Cho.

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Oh, Y.S., Kim, S.H. & Cho, GW. Functional Restoration of Amyotrophic Lateral Sclerosis Patient-Derived Mesenchymal Stromal Cells Through Inhibition of DNA Methyltransferase. Cell Mol Neurobiol 36, 613–620 (2016). https://doi.org/10.1007/s10571-015-0242-2

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