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Novel DNA aptamers that bind to mutant huntingtin and modify its activity

Shin, Baehyun and Jung, Roy and Oh, Hyejin and Owens, Gwen E. and Lee, Hyeongseok and Kwak, Seung and Lee, Ramee and Cotman, Susan L. and Lee, Jong-Min and MacDonald, Marcy E. and Song, Ji-Joon and Vijayvargia, Ravi and Seong, Ihn Sik (2018) Novel DNA aptamers that bind to mutant huntingtin and modify its activity. Molecular Therapy - Nucleic Acids, 11 . pp. 416-428. ISSN 2162-2531. PMCID PMC5992459. https://resolver.caltech.edu/CaltechAUTHORS:20180319-141833525

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Abstract

The CAG repeat expansion that elongates the polyglutamine tract in huntingtin is the root genetic cause of Huntington’s Disease (HD), a debilitating neurodegenerative disorder. This seemingly slight change to the primary amino acid sequence alters the physical structure of the mutant protein and alters its activity. We have identified a set of G-quadruplex-forming DNA aptamers (MS1, MS2, MS3, MS4) that bind mutant huntingtin proximal to lysines K2932/K2934 in the carboxyl-terminal CTD-II domain. Aptamer-binding to mutant huntingtin, abrogated the enhanced polycomb repressive complex 2 (PRC2) stimulatory-activity conferred by the expanded polyglutamine tract. In HD, but not normal, neuronal progenitor cells (NPC), MS3 aptamer co-localized with endogenous mutant huntingtin and was associated with significantly decreased PRC2 activity. Furthermore, MS3 transfection protected HD NPC against starvation-dependent stress with increased ATP. Therefore, DNA aptamers can preferentially target mutant huntingtin and modulate a gain of function endowed by the elongated polyglutamine segment. These mutant huntingtin binding aptamers provide novel molecular tools for delineating the effects of the HD mutation and encourage mutant huntingtin structure-based approaches to therapeutic development.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.omtn.2018.03.008DOIArticle
https://www.sciencedirect.com/science/article/pii/S2162253118300386PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5992459PubMed CentralArticle
Additional Information:© 2018 The Authors. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Received 28 December 2017, Accepted 13 March 2018, Available online 16 March 2018. We thank the members of the Cotman, Song, and Seong laboratories for suggestions and discussions. We are grateful to Dr. Ross Tomaino (the Taplin Mass spectrometry facility) for excellent technical support. This work was supported by the CHDI Foundation Inc [JML, MEM and ISS]; the National Institutes of Health/National Institute of Neurological Disorders and Stroke [R01 NS079651 to ISS]; and a Global Research Laboratory grant from National Foundation of Korea [NRF-2016K1A1A2912057 to ISS and JS]. Author Contributions: B.S., R.V., M.E.M. and I.S.S. developed the concept. B.S., R.J., H.O., G.E.O., H.L., S.L.C., J.M.L., J.S., R.V., S.K., R.L. and I.S.S. designed the experiments and analyzed the data. B.S., R.J. and R.V. produced and purified proteins. R.V. performed aptamer screening using LC science platform and ELISA validation experiments and R.V. and J.M.L. analyzed the data. G.E.O. performed ThT fluorescence assay. R.V. performed the aptamer binding experiment with caspase 6 cleavage. H.L., J.S. performed the aptamer binding experiment using SLM reaction and B.S. analyzed the data. R.J. and B.S. validated MS aptamer binding sites with KD mutants. R.V. performed huntingtin interaction and cell-free activity experiments with PRC2. B.S. performed the binding experiment of MS3 with NPC lysates and bead-based PRC2 assay. B.S., H.O. and S.L.C. performed immunofluorescence experiments and analyzed the confocal image data. B.S., R.J., H.L., J.S., R.V. and I.S.S. wrote the paper and significantly revised by S.K., R.L. and M.E.M. The authors declare no competing financial interests.
Funders:
Funding AgencyGrant Number
CHDI FoundationUNSPECIFIED
NIHR01 NS079651
National Research Foundation of KoreaNRF-2016K1A1A2912057
PubMed Central ID:PMC5992459
Record Number:CaltechAUTHORS:20180319-141833525
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180319-141833525
Official Citation:Baehyun Shin, Roy Jung, Hyejin Oh, Gwen E. Owens, Hyeongseok Lee, Seung Kwak, Ramee Lee, Susan L. Cotman, Jong-Min Lee, Marcy E. MacDonald, Ji-Joon Song, Ravi Vijayvargia, Ihn Sik Seong, Novel DNA Aptamers that Bind to Mutant Huntingtin and Modify Its Activity, Molecular Therapy - Nucleic Acids, Volume 11, 2018, Pages 416-428, ISSN 2162-2531, https://doi.org/10.1016/j.omtn.2018.03.008. (http://www.sciencedirect.com/science/article/pii/S2162253118300386)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:85364
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:26 Mar 2018 17:34
Last Modified:03 Oct 2019 19:30

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