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Targeting G-quadruplex DNA as cognitive function therapy for ATR-X syndrome

Nature Medicine volume 24, pages 802–813 (2018)Cite this article

Abstract

Alpha-thalassemia X-linked intellectual disability (ATR-X) syndrome is caused by mutations in ATRX, which encodes a chromatin-remodeling protein. Genome-wide analyses in mouse and human cells indicate that ATRX tends to bind to G-rich sequences with a high potential to form G-quadruplexes. Here, we report that Atrx mutation induces aberrant upregulation of Xlr3b expression in the mouse brain, an outcome associated with neuronal pathogenesis displayed by ATR-X model mice. We show that ATRX normally binds to G-quadruplexes in CpG islands of the imprinted Xlr3b gene, regulating its expression by recruiting DNA methyltransferases. Xlr3b binds to dendritic mRNAs, and its overexpression inhibits dendritic transport of the mRNA encoding CaMKII-α, promoting synaptic dysfunction. Notably, treatment with 5-ALA, which is converted into G-quadruplex-binding metabolites, reduces RNA polymerase II recruitment and represses Xlr3b transcription in ATR-X model mice. 5-ALA treatment also rescues decreased synaptic plasticity and cognitive deficits seen in ATR-X model mice. Our findings suggest a potential therapeutic strategy to target G-quadruplexes and decrease cognitive impairment associated with ATR-X syndrome.

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Fig. 1: ATRX regulates Xlr3b gene expression.
Fig. 2: Xlr3b is found in neuronal RNA granules.
Fig. 3: Aberrant Xlr3b expression perturbs synaptic plasticity by inhibiting dendritic Camk2a mRNA transport.
Fig. 4: 5-ALA represses Xlr3b transcription with Pol II recruitment by modifying the G-quadruplex structure.
Fig. 5: Treatment with 5-ALA counteracts cognitive deficits seen in AtrxΔE2 mice.

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Acknowledgements

We thank I. Kitajima for kindly providing AtrxΔE2 mice, deposited in the RIKEN BioResource Center (RBRC04937), H. Shimbo for kindly assisting in cell culture, D. Picketts for kindly providing the Atrx cDNA (pEGFP-C2-ATRX-HA) plasmid, N. Berube for kindly providing the ATRX shRNA (pSUPER-shATRX1) plasmid and K. Kosik for kindly providing the GFP-MS2-nls and MS2-binding site–CaMKII-α 3′ UTR plasmids. This research was supported by the Practical Research Project for Rare/Intractable Diseases from the Japan Agency for Medical Research and Development (AMED; N.S., K.K., H.T., N.O., H.S., K.F. and T.W.). This work was also supported by MEXT/JSPS KAKENHI (grant numbers 16K08265 and 25110705) to N.S.

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Authors and Affiliations

  1. Department of Biofunctional Analysis Laboratory of Molecular Biology, Gifu Pharmaceutical University, Gifu, Japan

    Norifumi Shioda

  2. Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan

    Yasushi Yabuki, Kouya Yamaguchi, Misaki Onozato & Kohji Fukunaga

  3. Department of Chemistry, Graduate School of Science, Kyoto University, Kyoto, Japan

    Yue Li & Hiroshi Sugiyama

  4. Division of Genetics, Kanagawa Children’s Medical Center, Yokohama, Japan

    Kenji Kurosawa

  5. Department of Evolutionary Studies of Biosystems, School of Advanced Sciences, SOKENDAI (The Graduate University for Advanced Studies), Hayama, Japan

    Hideyuki Tanabe

  6. Department of Medical Genetics, Osaka Women’s and Children’s Hospital, Osaka, Japan

    Nobuhiko Okamoto

  7. Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan

    Takumi Era

  8. Department of Medical Ethics and Medical Genetics, Graduate School of Medicine, Kyoto University, Kyoto, Japan

    Takahito Wada

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  1. Norifumi Shioda
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Contributions

N.S., Y.Y., K.Y., M.O. and Y.L. performed the experiments. K.K., H.T., N.O., T.E., H.S. and T.W. provided critical advice. N.S. and K.F. wrote the manuscript and designed the study. All authors discussed the results and commented on the manuscript.

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Correspondence to Norifumi Shioda, Takahito Wada or Kohji Fukunaga.

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Supplementary Text and Figures

Supplementary Figures 1–13 and Supplementary Tables 1–5

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Supplementary Video 1

Time-lapse imaging of GFP-MS2-labeled CaMKIIα mRNA (GFP-CaMKIIα 3′ UTR) in a proximal dendrite of a cultured WT neuron

Supplementary Video 2

Time-lapse imaging of GFP-MS2-labeled CaMKIIα mRNA (GFP-CaMKIIα 3′ UTR) in a distal dendrite of a cultured WT neuron

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Shioda, N., Yabuki, Y., Yamaguchi, K. et al. Targeting G-quadruplex DNA as cognitive function therapy for ATR-X syndrome. Nat Med 24, 802–813 (2018). https://doi.org/10.1038/s41591-018-0018-6

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