Nuclear Localization of Huntingtin mRNA Is Specific to Cells of Neuronal Origin
Open accessAbstract
Huntington's disease (HD) is a monogenic neurodegenerative disorder representing an ideal candidate for gene silencing with oligonucleotide therapeutics (i.e., antisense oligonucleotides [ASOs] and small interfering RNAs [siRNAs]). Using an ultra-sensitive branched fluorescence in situ hybridization (FISH) method, we show that ∼50% of wild-type HTT mRNA localizes to the nucleus and that its nuclear localization is observed only in neuronal cells. In mouse brain sections, we detect Htt mRNA predominantly in neurons, with a wide range of Htt foci observed per cell. We further show that siRNAs and ASOs efficiently eliminate cytoplasmic HTT mRNA and HTT protein, but only ASOs induce a partial but significant reduction of nuclear HTT mRNA. We speculate that, like other mRNAs, HTT mRNA subcellular localization might play a role in important neuronal regulatory mechanisms.
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A fraction of Htt mRNA localizes to the nucleus, making it more resistant to RNAi-based silencing.
“The extent of HTT protein silencing was always greater than Htt mRNA (> 99% vs ~70%) (Fig. 2c,d) because a fraction of Htt mRNA localizes to the nucleus, making it more resistant to RNAi-based silencing29.”
From A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system (2019) · cites this paper as [29]
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References 24 · Cited by 64
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- Alterman JF, Coles AH, Hall LM, Aronin N, Khvorova A, and Didiot MC (2017). A high-throughput assay for mRNA silencing in primary cortical neurons in vitro with oligonucleotide therapeutics. Biol. Protoc. 7, e2501. PMC5621760 28966945
- Blair JD, Hockemeyer D, Doudna JA, Bateup HS, and Floor SN (2017). Widespread translational remodeling during human neuronal differentiation. Cell Rep. 21, 2005– 2016. 29141229 PMC5759054
- Bolte S, and Cordelières FP (2006). A guided tour into subcellular colocalization analysis in light microscopy. J. Microsc. 224, 213– 232. 17210054
- Castanotto D, Lin M, Kowolik C, Wang L, Ren XQ, Soifer HS, Koch T, Hansen BR, Oerum H, Armstrong B, (2015). A cytoplasmic pathway for gapmer antisense oligonucleotide-mediated gene silencing in mammalian cells. Nucleic Acids Res. 43, 9350– 9361. 26433227 PMC4627093
- Crooke ST, Witztum JL, Bennett CF, and Baker BF (2018). RNA- targeted therapeutics. Cell Metab. 27, 714– 739. 29617640
- de Mezer M, Wojciechowska M, Napierala M, Sobczak K, and Krzyzo- siak WJ (2011). Mutant CAG repeats of Huntingtin transcript fold into hairpins, form nuclear foci and are targets for RNA interference. Nucleic Acids Res. 39, 3852– 3863. 21247881 PMC3089464
- Keeler AM, Sapp E, Chase K, Sottosanti E, Danielson E, Pfister E, Stoica L, DiFiglia M, Aronin N, and Sena-Esteves M (2016). Cellular analysis of silencing the Huntington’s disease gene using AAV9 mediated delivery of artificial micro RNA into the striatum of Q140/Q140 mice. J. Huntingtons Dis. 5, 239– 248. 27689620
- Kordasiewicz HB, Stanek LM, Wancewicz EV, Mazur C, McAlonis MM, Pytel KA, Artates JW, Weiss A, Cheng SH, Shihabuddin LS, (2012). Sustained therapeutic reversal of Huntington’s disease by transient repression of huntingtin synthesis. Neuron 74, 1031– 1044. 22726834 PMC3383626
- Landwehrmeyer GB, McNeil SM, Dure LS 4th, Ge P, Aizawa H, Huang Q, Ambrose CM, Duyao MP, Bird ED, Bonilla E, (1995). Huntington’s disease gene: regional and cellular expression in brain of normal and affected individuals. Ann. Neurol. 37, 218– 230. 7847863
- Ollion J, Cochennec J, Loll F, Escude C, and Boudier T (2013). TANGO: a generic tool for high-throughput 3D image analysis for studying nuclear organization. Bioinformatics 29, 1840– 1841. 23681123 PMC3702251
- Otsu N (1979). Athreshold selection method from gray-level histograms. IEEE Trans. Syst. Man Cybern. 9, 62– 66.
- Patange S, Girvan M, and Larson DR (2018). Single-cell systems biology: probing the basic unit of information flow. Curr. Opin. Syst. Biol. 8, 7– 15. 29552672 PMC5851483
- Richner M, Victor MB, Liu Y, Abernathy D, and Yoo AS (2015). MicroRNA-based conversion of human fibroblasts into striatal medium spiny neurons. Nat. Protoc. 70, 1543– 1555. PMC12176329 26379228
- Sapp E, Valencia A, Li X, Aronin N, Kegel KB, Vonsattel JP, Young AB, Wexler N, and DiFiglia M (2012). Native mutant huntingtin in human brain: evidence for prevalence of full-length monomer. J. Biol. Chem. 287, 13487– 13499. 22375012 PMC3339996
- Schneider CA, Rasband WS, and Eliceiri KW (2012). NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671– 675. 22930834 PMC5554542
- Seluanov A, Vaidya A, and Gorbunova V (2010). Establishing primary adult fibroblast cultures from rodents. J. Vis. Exp, 2033. PMC3185624 20972406
- Tang J, Yoo AS, and Crabtree GR (2013). Reprogramming human fibroblasts to neurons by recapitulating an essential microRNA-chromatin switch. Curr. Opin. Genet. Dev. 23, 591– 598. 24035011 PMC3951511
- The Huntington’s Disease Collaborative Research Group (1993). A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell 72, 971– 983. 8458085
- Urbanek MO, Fiszer A, and Krzyzosiak WJ (2017). Reduction of Huntington’s disease RNA foci by CAG repeat-targeting reagents. Front. Cell. Neurosci. 11, 82. 28400719 PMC5368221
- Wang F, Flanagan J, Su N, Wang LC, Bui S, Nielson A, Wu X, Vo HT, Ma XJ, and Luo Y (2012). RNAscope: a novel in situ RNA analysis platform for formalin-fixed, paraffin-embedded tissues. J. Mol. Diagn. 14, 22– 29. 22166544 PMC3338343
- Zack GW, Rogers WE, and Latt SA (1977). Automatic measurement of sister chromatid exchange frequency. J. Histochem. Cytochem. 25, 741– 753. 70454
- Authors
- Didiot, M.-C., Ferguson, C. M., Ly, S., Coles, A. H., Smith, A. O., Bicknell, A. A., Hall, L. M., Sapp, E., Echeverria, D., Pai, A. A., DiFiglia, M., Moore, M. J., Hayward, L. J., Aronin, N., & Khvorova, A.
- Affiliations
- University of Massachusetts Chan Medical School (US)
- Massachusetts General Hospital (US)
- Corresponding author
- Didiot, Marie-Cécile
- Khvorova, Anastasia
- ORCID
- Venue
- Cell Reports
- Publisher
- Elsevier BV
- Published
- 2018-09
- Volume
- 24
- Issue
- 10
- Pages
- 2553-2560.e5
- ISSN
- 2211-1247 (ISSN-L)
- PMC
- PMC6245656
- OpenAlex
- W2891861470
- Cited by
- 64
- References
- 24
- Citations by year
- 2026: 6 · 2025: 5 · 2024: 10 · 2023: 21 · 2022: 9 · 2021: 3 · 2020: 6 · 2019: 3 · 2017: 1
- FWCI
- 1.37
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Cell Reports (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- The Journal of the American Medical Association (JAMA) Network (American Medical Association) (submittedVersion, cc-by-nc-nd)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- NIH (R01 NS104022-01)
- CHDI Foundation (A-5038)
- HDSA
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Genetic Neurodegenerative Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Keywords
- Huntington's disease, neurons, antisense oligonucleotides, siRNA, gene silencing, subcellular mRNA localization, mouse brain
- MeSH terms
- Animals, Cell Nucleus, Cells, Cultured, Female, HeLa Cells, Humans, Huntington Disease, Neurons, RNA, Messenger, Oligonucleotides, Antisense, In Situ Hybridization, Fluorescence, Trinucleotide Repeat Expansion, Gene Silencing, RNA, Small Interfering, Mice
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