Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease

Edith L. Pfister; Natalie Dinardo; Erica Mondo; Florie Borel; Faith Conroy; Cara Fraser; Gwladys Gernoux; Xin Han; Danjing Hu; Emily Johnson; Lori Kennington; Pengpeng Liu; Suzanne J. Reid; Ellen Sapp; Petr Vodicka; Tim Kuchel; A. Jennifer Morton; David Howland; Richard Moser; Miguel Sena-Esteves; Guangping Gao; Christian Mueller; Marian Difiglia; Neil Aronin

doi:10.1089/hum.2017.199

Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease

Edith L. Pfister, Natalie Dinardo, Erica Mondo, Florie Borel, Faith Conroy, Cara Fraser, Gwladys Gernoux, Xin Han, Danjing Hu, Emily Johnson, Lori Kennington, Pengpeng Liu, Suzanne J. Reid, Ellen Sapp, Petr Vodicka, Tim Kuchel, A. Jennifer Morton, David Howland, Richard Moser, Miguel Sena-EstevesGuangping Gao, Christian Mueller, Marian Difiglia, Neil Aronin

Preclinical Imaging And Research Laboratories

Research output: Contribution to journal › Article › peer-review

72 Citations (Scopus)

Abstract

Huntington's disease (HD) is a fatal neurodegenerative disease caused by a genetic expansion of the CAG repeat region in the huntingtin (HTT) gene. Studies in HD mouse models have shown that artificial miRNAs can reduce mutant HTT, but evidence for their effectiveness and safety in larger animals is lacking. HD transgenic sheep express the full-length human HTT with 73 CAG repeats. AAV9 was used to deliver unilaterally to HD sheep striatum an artificial miRNA targeting exon 48 of the human HTT mRNA under control of two alternative promoters: U6 or CβA. The treatment reduced human mutant (m) HTT mRNA and protein 50-80% in the striatum at 1 and 6 months post injection. Silencing was detectable in both the caudate and putamen. Levels of endogenous sheep HTT protein were not affected. There was no significant loss of neurons labeled by DARPP32 or NeuN at 6 months after treatment, and Iba1-positive microglia were detected at control levels. It is concluded that safe and effective silencing of human mHTT protein can be achieved and sustained in a large-animal brain by direct delivery of an AAV carrying an artificial miRNA.

Original language	English
Pages (from-to)	663-673
Number of pages	11
Journal	Human Gene Therapy
Volume	29
Issue number	6
DOIs	https://doi.org/10.1089/hum.2017.199
Publication status	Published or Issued - Jun 2018

Keywords

AAV
Huntington's disease
RNAi
large animal models

ASJC Scopus subject areas

Molecular Medicine
Molecular Biology
Genetics

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10.1089/hum.2017.199

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Pfister, E. L., Dinardo, N., Mondo, E., Borel, F., Conroy, F., Fraser, C., Gernoux, G., Han, X., Hu, D., Johnson, E., Kennington, L., Liu, P., Reid, S. J., Sapp, E., Vodicka, P., Kuchel, T., Morton, A. J., Howland, D., Moser, R., ... Aronin, N. (2018). Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease. Human Gene Therapy, 29(6), 663-673. https://doi.org/10.1089/hum.2017.199

@article{29d05b8d97da4e13af712d673183ed41,

title = "Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease",

abstract = "Huntington's disease (HD) is a fatal neurodegenerative disease caused by a genetic expansion of the CAG repeat region in the huntingtin (HTT) gene. Studies in HD mouse models have shown that artificial miRNAs can reduce mutant HTT, but evidence for their effectiveness and safety in larger animals is lacking. HD transgenic sheep express the full-length human HTT with 73 CAG repeats. AAV9 was used to deliver unilaterally to HD sheep striatum an artificial miRNA targeting exon 48 of the human HTT mRNA under control of two alternative promoters: U6 or CβA. The treatment reduced human mutant (m) HTT mRNA and protein 50-80% in the striatum at 1 and 6 months post injection. Silencing was detectable in both the caudate and putamen. Levels of endogenous sheep HTT protein were not affected. There was no significant loss of neurons labeled by DARPP32 or NeuN at 6 months after treatment, and Iba1-positive microglia were detected at control levels. It is concluded that safe and effective silencing of human mHTT protein can be achieved and sustained in a large-animal brain by direct delivery of an AAV carrying an artificial miRNA.",

keywords = "AAV, Huntington's disease, RNAi, large animal models",

author = "Pfister, {Edith L.} and Natalie Dinardo and Erica Mondo and Florie Borel and Faith Conroy and Cara Fraser and Gwladys Gernoux and Xin Han and Danjing Hu and Emily Johnson and Lori Kennington and Pengpeng Liu and Reid, {Suzanne J.} and Ellen Sapp and Petr Vodicka and Tim Kuchel and Morton, {A. Jennifer} and David Howland and Richard Moser and Miguel Sena-Esteves and Guangping Gao and Christian Mueller and Marian Difiglia and Neil Aronin",

year = "2018",

month = jun,

doi = "10.1089/hum.2017.199",

language = "English",

volume = "29",

pages = "663--673",

journal = "Human Gene Therapy",

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Pfister, EL, Dinardo, N, Mondo, E, Borel, F, Conroy, F, Fraser, C, Gernoux, G, Han, X, Hu, D, Johnson, E, Kennington, L, Liu, P, Reid, SJ, Sapp, E, Vodicka, P, Kuchel, T, Morton, AJ, Howland, D, Moser, R, Sena-Esteves, M, Gao, G, Mueller, C, Difiglia, M & Aronin, N 2018, 'Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease', Human Gene Therapy, vol. 29, no. 6, pp. 663-673. https://doi.org/10.1089/hum.2017.199

TY - JOUR

T1 - Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease

AU - Pfister, Edith L.

AU - Dinardo, Natalie

AU - Mondo, Erica

AU - Borel, Florie

AU - Conroy, Faith

AU - Fraser, Cara

AU - Gernoux, Gwladys

AU - Han, Xin

AU - Hu, Danjing

AU - Johnson, Emily

AU - Kennington, Lori

AU - Liu, Pengpeng

AU - Reid, Suzanne J.

AU - Sapp, Ellen

AU - Vodicka, Petr

AU - Kuchel, Tim

AU - Morton, A. Jennifer

AU - Howland, David

AU - Moser, Richard

AU - Sena-Esteves, Miguel

AU - Gao, Guangping

AU - Mueller, Christian

AU - Difiglia, Marian

AU - Aronin, Neil

PY - 2018/6

Y1 - 2018/6

N2 - Huntington's disease (HD) is a fatal neurodegenerative disease caused by a genetic expansion of the CAG repeat region in the huntingtin (HTT) gene. Studies in HD mouse models have shown that artificial miRNAs can reduce mutant HTT, but evidence for their effectiveness and safety in larger animals is lacking. HD transgenic sheep express the full-length human HTT with 73 CAG repeats. AAV9 was used to deliver unilaterally to HD sheep striatum an artificial miRNA targeting exon 48 of the human HTT mRNA under control of two alternative promoters: U6 or CβA. The treatment reduced human mutant (m) HTT mRNA and protein 50-80% in the striatum at 1 and 6 months post injection. Silencing was detectable in both the caudate and putamen. Levels of endogenous sheep HTT protein were not affected. There was no significant loss of neurons labeled by DARPP32 or NeuN at 6 months after treatment, and Iba1-positive microglia were detected at control levels. It is concluded that safe and effective silencing of human mHTT protein can be achieved and sustained in a large-animal brain by direct delivery of an AAV carrying an artificial miRNA.

AB - Huntington's disease (HD) is a fatal neurodegenerative disease caused by a genetic expansion of the CAG repeat region in the huntingtin (HTT) gene. Studies in HD mouse models have shown that artificial miRNAs can reduce mutant HTT, but evidence for their effectiveness and safety in larger animals is lacking. HD transgenic sheep express the full-length human HTT with 73 CAG repeats. AAV9 was used to deliver unilaterally to HD sheep striatum an artificial miRNA targeting exon 48 of the human HTT mRNA under control of two alternative promoters: U6 or CβA. The treatment reduced human mutant (m) HTT mRNA and protein 50-80% in the striatum at 1 and 6 months post injection. Silencing was detectable in both the caudate and putamen. Levels of endogenous sheep HTT protein were not affected. There was no significant loss of neurons labeled by DARPP32 or NeuN at 6 months after treatment, and Iba1-positive microglia were detected at control levels. It is concluded that safe and effective silencing of human mHTT protein can be achieved and sustained in a large-animal brain by direct delivery of an AAV carrying an artificial miRNA.

KW - AAV

KW - Huntington's disease

KW - RNAi

KW - large animal models

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U2 - 10.1089/hum.2017.199

DO - 10.1089/hum.2017.199

M3 - Article

C2 - 29207890

AN - SCOPUS:85044582983

SN - 1043-0342

VL - 29

SP - 663

EP - 673

JO - Human Gene Therapy

JF - Human Gene Therapy

IS - 6

ER -

Artificial miRNAs Reduce Human Mutant Huntingtin Throughout the Striatum in a Transgenic Sheep Model of Huntington's Disease

Abstract

Keywords

ASJC Scopus subject areas

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