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Developmental alterations in Huntington's disease neural cells and pharmacological rescue in cells and mice

Nature Neuroscience volume 20, pages 648660 (2017) | Download Citation

Abstract

Neural cultures derived from Huntington's disease (HD) patient-derived induced pluripotent stem cells were used for 'omics' analyses to identify mechanisms underlying neurodegeneration. RNA-seq analysis identified genes in glutamate and GABA signaling, axonal guidance and calcium influx whose expression was decreased in HD cultures. One-third of gene changes were in pathways regulating neuronal development and maturation. When mapped to stages of mouse striatal development, the profiles aligned with earlier embryonic stages of neuronal differentiation. We observed a strong correlation between HD-related histone marks, gene expression and unique peak profiles associated with dysregulated genes, suggesting a coordinated epigenetic program. Treatment with isoxazole-9, which targets key dysregulated pathways, led to amelioration of expanded polyglutamine repeat-associated phenotypes in neural cells and of cognitive impairment and synaptic pathology in HD model R6/2 mice. These data suggest that mutant huntingtin impairs neurodevelopmental pathways that could disrupt synaptic homeostasis and increase vulnerability to the pathologic consequence of expanded polyglutamine repeats over time.

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Acknowledgements

We thank the patients and their families for their essential contributions to this research. We also thank E. Cattaneo and J. Arjomond for discussions of the data, D. Merry for critique of the manuscript and data, S. Svendsen for editorial assistance, J. Dunn for technical culture assistance, G. Vatine (Cedars-Sinai Medical Center, Los Angeles) for the iPSC-derived oligodendrocyte precursors, M. Godoy and G. Gowing (Cedars-Sinai Medical Center, Los Angeles) for the rat muscle and R. Barrett (Cedars-Sinai Medical Center, Los Angeles) for the definitive endoderm positive control. We also thank F. Bennet and Ionis Pharmaceuticals for providing the HTT ASO. Primary support for this work was from NIH NS078370 (L.M.T., C.N.S., J.F.G., M.E.M., C.A.R. and S.F.) and from the CHDI Foundation (J.M.C, P.J.K. and N.D.A.). Additional support was provided by NIH: U54 NS091046 NeuroLINCS center (L.M.T., C.N.S., E.F.); NIH NS089076 (L.M.T., D.E.H., E.F.); P50NS16367 (HD Center Without Walls); NIH R01GM089903 (E.F.); NIH NS101996-01 (S.F.); NIH R01NS084298 (B.S.); American Heart Association, CIRM and NRSA fellowships (R.G.L.); the Hereditary Disease Foundation (V.B.M.); the Taube-Koret Center and the Hellman Family Foundation (S.F.); the UCI Institute for Clinical and Translational Science (L.M.T.); Huntington's Disease Society of America (L.L.S); HD CARE (L.M.T.) and the NIH Biotechnology Training Program Fellowship (T32GM008334, A.J.K.). Additional support was provided by grants from the Ministerio de Economia y Competitividad (SAF 2014-57160-R to JA; SAF2015-66505-R to J.M.C.), from the ISCIII-Subdirección General de Evaluación and European Regional Development Fund (ERDF) (RETICS to JMC (RD12/0019/0002; Red de Terapia Celular); ADVANCE(CAT) with the support of ACCIÓ (Catalonia Trade & Investment; Generalitat de Catalunya) and the European Community under the Catalonian ERDF operational program 2014-2020), Spain, from the European Union FP7 (P.J.K. and N.D.A.) and from the Ser Cymru Life Sciences & Health Network in Drug Discovery Programme (M.W.S.). This work was made possible, in part, through access to the Genomic High Throughput Facility Shared Resource of the Cancer Center Support Grant (CA-62203) at the University of California, Irvine. Support also included computing resources from National Science Foundation grant DB1-0821391 and sequencing support from National Institutes of Health grant P30-ES002109.

Author information

Author notes

Affiliations

  1. Department of Biological Chemistry, University of California, Irvine, Irvine, California, USA.

    • Ryan G Lim
    •  & Leslie M Thompson
  2. Department of Psychiatry and Human Behavior, University of California, Irvine, Irvine, California, USA.

    • Lisa L Salazar
    •  & Leslie M Thompson
  3. F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

    • Daniel K Wilton
    •  & Beth Stevens
  4. Department of Neurobiology and Behavior, University of California, Irvine, Irvine, California, USA.

    • Alvin R King
    • , Delaram Sharifabad
    • , Sara T Winokur
    •  & Leslie M Thompson
  5. UCI MIND, University of California, Irvine, Irvine, California, USA.

    • Jennifer T Stocksdale
    • , Alice L Lau
    • , Jack C Reidling
    • , Malcolm S Casale
    •  & Leslie M Thompson
  6. Sue and Bill Gross Stem Cell Center, University of California, Irvine, Irvine, California, USA.

    • Leslie M Thompson
  7. Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona Spain; August Pi i Sunyer Biomedical Research Institute (IDIBAPS), Barcelona, Spain; and Networked Biomedical Research Centre for NeuroDegenerative Disorders (CIBERNED), Barcelona, Spain.

    • Mónica Pardo
    • , A Gerardo García Díaz-Barriga
    • , Marco Straccia
    • , Phil Sanders
    • , Jordi Alberch
    •  & Josep M Canals
  8. Gladstone Institutes and the Taube/Koret Center of Neurodegenerative Disease Research, Roddenberry Stem Cell Research Program, San Francisco, California, USA.

    • Julia A Kaye
    • , Mariah Dunlap
    • , Lisa Jo
    • , Hanna May
    • , Elliot Mount
    • , Kelly Haston
    •  & Steven Finkbeiner
  9. Sandia National Laboratories, Livermore, California, USA.

    • Cliff Anderson-Bergman
  10. Departments of Neurology and Physiology, University of California, San Francisco, San Francisco, California, USA.

    • Steven Finkbeiner
  11. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Amanda J Kedaigle
    • , Ferah Yildirim
    • , Pamela Milani
    •  & Ernest Fraenkel
  12. Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Theresa A Gipson
    • , Christopher W Ng
    •  & David E Housman
  13. Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Theresa A Gipson
    • , Christopher W Ng
    •  & David E Housman
  14. School of Biomedical Sciences, Cardiff University, Cardiff, UK.

    • Nicholas D Allen
    •  & Paul J Kemp
  15. Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.

    • Ranjit Singh Atwal
    • , Marta Biagioli
    • , James F Gusella
    •  & Marcy E MacDonald
  16. Division of Neurobiology, Department of Psychiatry, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.

    • Sergey S Akimov
    • , Nicolas Arbez
    • , Jacqueline Stewart
    •  & Christopher A Ross
  17. Departments of Neurology, Neuroscience and Pharmacology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.

    • Christopher A Ross
  18. The Board of Governors Regenerative Medicine Institute and Department of Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, California, USA.

    • Virginia B Mattis
    • , Colton M Tom
    • , Loren Ornelas
    • , Anais Sahabian
    • , Lindsay Lenaeus
    • , Berhan Mandefro
    • , Dhruv Sareen
    •  & Clive N Svendsen

Consortia

  1. The HD iPSC Consortium

Authors

    Contributions

    Designed the experiments: R.G.L., L.L.S., D.K.W., J.C.R., S.T.W., L.M.T., J.A., J.M.C., N.D.A., P.J.K., J.A.K., S.F., F.Y., D.E.H., E.F., J.F.G., M.E.M., S.S.A., N.A., C.A.R., V.B.M. and C.N.S. Generated iPSC lines in study: L.O., A.S., L.L., B.M. and D. Sareen. iPSC culture and neuronal differentiation: V.B.M., L.L.S., A.R.K., J.T.S., C.M.T., S.S.A., J.A.K., H.M. and M.D. Carried out experiments: R.G.L. and T.A.G., RNA-seq; L.L.S., Isx-9 qPCR and NEUROD1 overexpression; A.L.L., M.P., A.G.G.D.-B., M.S. and P.S., mouse neurodevelopment studies; A.G.G.D.-B., comparison between mouse and human data; V.B.M. and C.M.T., cell counts; V.B.M., immunocytochemistry; F.Y., R.S.A. and M.B., ChIP; S.S.A., Cell Titer-Glo cell survival assay; S.S.A., N.A. and L.L.S., NEUROD1 knockdown; N.A. and J.S., cell culture and transfection of mouse primary neurons and nuclear condensation assay; S.S.A., Western analysis; E.M., J.A.K., M.D. and H.M., Isx-9 neuron assays; D.K.W., Isx-9 synaptic assays; J.C.R. and D. Sharifabad, mouse Isx-9 studies. Analyzed the data: R.G.L., L.L.S., D.K.W., B.S., J.C.R., M.S.C., S.T.W., L.M.T., J.A.K., M.D., H.M., L.J., D.K.W., C.A.-B., S.F., A.J.K., T.A.G., F.Y., C.W.N., P.M., D.E.H., E.F., J.F.G., M.E.M., S.S.A., N.A., C.A.R., V.B.M. and C.N.S. Wrote the manuscript: R.G.L., L.L.S., D.K.W., J.C.R., S.T.W., L.M.T., J.M.C., N.D.A., P.J.K., J.A.K., K.H., S.F., A.J.K., T.A.G., P.M., D.E.H., E.F., M.E.M., J.F.G., S.S.A., C.A.R., V.B.M. and C.N.S. A list of authors by individual consortium group appears in the Supplementary Note.

    Competing interests

    The author declare no competing financial interests.

    Corresponding author

    Correspondence to Leslie M Thompson.

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    https://doi.org/10.1038/nn.4532

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