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Choline Ameliorates Disease Phenotypes in Human iPSC Models of Rett Syndrome

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Abstract

Rett syndrome (RTT) is a postnatal neurodevelopmental disorder that primarily affects girls. Mutations in the methyl-CpG-binding protein 2 (MECP2) gene account for approximately 95 % of all RTT cases. To model RTT in vitro, we generated induced pluripotent stem cells (iPSCs) from fibroblasts of two RTT patients with different mutations (MECP2 R306C and MECP2 1155Δ32) in their MECP2 gene. We found that these iPSCs were capable of differentiating into functional neurons. Compared to control neurons, the RTT iPSC-derived cells had reduced soma size and a decreased amount of synaptic input, evident both as fewer Synapsin 1-positive puncta and a lower frequency of spontaneous excitatory postsynaptic currents. Supplementation of the culture media with choline rescued all of these defects. Choline supplementation may act through changes in the expression of choline acetyltransferase, an important enzyme in cholinergic signaling, and also through alterations in the lipid metabolite profiles of the RTT neurons. Our study elucidates the possible mechanistic pathways for the effect of choline on human RTT cell models, thereby illustrating the potential for using choline as a nutraceutical to treat RTT.

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Acknowledgments

We thank W. Y. Leong for technical support and members of the Goh and Augustine labs for sharing reagents and expertise, and J. Ching and K. T. Fridianto from the Duke-NUS Metabolomics Facility for processing the samples and assistance in curating the data for lipid metabolomics.

Funding

This work was primarily supported by Abbott Nutrition, Cognition Center of Excellence, Singapore R&D, and also partially supported by the Academic Center of Excellence (ACE) research award from GlaxoSmithKline (GSK), the National Research Foundation Singapore under its Competitive Research Program (NRF 2008 NRF-CRP 002-082), the National Research Medical Council (NMRC)—Collaborative Research Programme Grant (CBRG)—(NMRC/CBRG/0094/2015) and the Ministry of Education (MOE) TiER 2 (MOE2015-T2-1-022) to EG.

Author Contributions

EC characterized the iPSC-derived NPCs and neurons, performed lipid-profiling experiments and analysis, Western blots, and analysis for ChAT, prepared all figures, and wrote the manuscript. GM reprogrammed and characterized the iPSCs and the iPSC-derived NPCs and neurons. SY did all electrophysiological recordings and analyses. DM characterized differentiation properties of iPSCs and did Western blots and analysis for ChAT. FR provided critical inputs to the design of the project. GA designed electrophysiology experiments and provided critical input to data analysis and manuscript. EG initiated and directed the entire study, designed experiments, analyzed data, and wrote the manuscript.

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    Authors and Affiliations

    1. Neuroscience Academic Clinical Programme, Duke-NUS Medical School, 8 College Road, Singapore, 169857, Singapore

      Eunice W. M. Chin, Guillaume Marcy, Dongliang Ma & Eyleen L. K. Goh

    2. Department of Research, National Neuroscience Institute, Singapore, 308433, Singapore

      Eunice W. M. Chin, Dongliang Ma & Eyleen L. K. Goh

    3. NUS Graduate School of Integrative Sciences and Engineering, National University of Singapore, Singapore, 117597, Singapore

      Eunice W. M. Chin

    4. Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore, Singapore

      Su-In Yoon & George J. Augustine

    5. Abbott Nutrition Research and Development, Columbus, Ohio, USA

      Francisco J. Rosales

    6. Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117597, Singapore

      Eyleen L. K. Goh

    7. KK Research Center, KK Women’s and Children’s Hospital, Singapore, 229899, Singapore

      Eyleen L. K. Goh

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    1. Eunice W. M. Chin
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    Correspondence to George J. Augustine or Eyleen L. K. Goh.

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    Eunice W. M. Chin, Guillaume Marcy, Su-In Yoon have contributed equally to the work.

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    Chin, E.W.M., Marcy, G., Yoon, SI. et al. Choline Ameliorates Disease Phenotypes in Human iPSC Models of Rett Syndrome. Neuromol Med 18, 364–377 (2016). https://doi.org/10.1007/s12017-016-8421-y

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