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Akimov SS, Jiang M, Kedaigle AJ, et al. Immortalized striatal precursor neurons from Huntington's disease patient-derived iPS cells as a platform for target identification and screening for experimental therapeutics. Hum Mol Genet. 2021;30(24):2469-2487doi: 10.1093/hmg/ddab200.
Akimov, S. S., Jiang, M., Kedaigle, A. J., Arbez, N., Marque, L. O., Eddings, C. R., Ranum, P. T., Whelan, E., Tang, A., Wang, R., DeVine, L. R., Talbot, C. C., Cole, R. N., Ratovitski, T., Davidson, B. L., Fraenkel, E., & Ross, C. A. (2021). Immortalized striatal precursor neurons from Huntington's disease patient-derived iPS cells as a platform for target identification and screening for experimental therapeutics. Human molecular genetics, 30(24), 2469-2487. https://doi.org/10.1093/hmg/ddab200
Akimov, Sergey S, et al. "Immortalized striatal precursor neurons from Huntington's disease patient-derived iPS cells as a platform for target identification and screening for experimental therapeutics." Human molecular genetics vol. 30,24 (2021): 2469-2487. doi: https://doi.org/10.1093/hmg/ddab200
Akimov SS, Jiang M, Kedaigle AJ, Arbez N, Marque LO, Eddings CR, Ranum PT, Whelan E, Tang A, Wang R, DeVine LR, Talbot CC, Cole RN, Ratovitski T, Davidson BL, Fraenkel E, Ross CA. Immortalized striatal precursor neurons from Huntington's disease patient-derived iPS cells as a platform for target identification and screening for experimental therapeutics. Hum Mol Genet. 2021 Nov 30;30(24):2469-2487. doi: 10.1093/hmg/ddab200. PMID: 34296279; PMCID: PMC8643509.
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Hum Mol Genet. 2021 Nov 30;30(24):2469-2487. doi: 10.1093/hmg/ddab200.

Immortalized striatal precursor neurons from Huntington's disease patient-derived iPS cells as a platform for target identification and screening for experimental therapeutics.

Human molecular genetics

Sergey S Akimov, Mali Jiang, Amanda J Kedaigle, Nicolas Arbez, Leonard O Marque, Chelsy R Eddings, Paul T Ranum, Emma Whelan, Anthony Tang, Ronald Wang, Lauren R DeVine, Conover C Talbot, Robert N Cole, Tamara Ratovitski, Beverly L Davidson, Ernest Fraenkel, Christopher A Ross

Affiliations

  1. Division of Neurobiology, Department of Psychiatry, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
  2. Department of Biological Engineering, Computational and Systems Biology Graduate Program, Massachusetts Institute of Technology, Cambridge, MA 02142, USA.
  3. The Department of Pathology and Laboratory Medicine, The University of Pennsylvania, The Raymond G Perelman Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
  4. Mass Spectrometry and Proteomics Facility, Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
  5. The Johns Hopkins School of Medicine, Institute for Basic Biomedical Sciences, Baltimore, MD 21205, USA.
  6. The Department of Pathology and Laboratory Medicine, The University of Pennsylvania, Philadelphia, PA 19104, USA.
  7. Department of Neurology, Neuroscience and Pharmacology, The Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.

PMID: 34296279 PMCID: PMC8643509 DOI: 10.1093/hmg/ddab200

Abstract

We have previously established induced pluripotent stem cell (iPSC) models of Huntington's disease (HD), demonstrating CAG-repeat-expansion-dependent cell biological changes and toxicity. However, the current differentiation protocols are cumbersome and time consuming, making preparation of large quantities of cells for biochemical or screening assays difficult. Here, we report the generation of immortalized striatal precursor neurons (ISPNs) with normal (33) and expanded (180) CAG repeats from HD iPSCs, differentiated to a phenotype resembling medium spiny neurons (MSN), as a proof of principle for a more tractable patient-derived cell model. For immortalization, we used co-expression of the enzymatic component of telomerase hTERT and conditional expression of c-Myc. ISPNs can be propagated as stable adherent cell lines, and rapidly differentiated into highly homogeneous MSN-like cultures within 2 weeks, as demonstrated by immunocytochemical criteria. Differentiated ISPNs recapitulate major HD-related phenotypes of the parental iPSC model, including brain-derived neurotrophic factor (BDNF)-withdrawal-induced cell death that can be rescued by small molecules previously validated in the parental iPSC model. Proteome and RNA-seq analyses demonstrate separation of HD versus control samples by principal component analysis. We identified several networks, pathways, and upstream regulators, also found altered in HD iPSCs, other HD models, and HD patient samples. HD ISPN lines may be useful for studying HD-related cellular pathogenesis, and for use as a platform for HD target identification and screening experimental therapeutics. The described approach for generation of ISPNs from differentiated patient-derived iPSCs could be applied to a larger allelic series of HD cell lines, and to comparable modeling of other genetic disorders.

© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For Permissions, please email: [email protected].

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