A high-content drug screening strategy to identify protein level modulators for genetic diseases: A proof-of-principle in autosomal dominant leukodystrophy.


Journal

Human mutation
ISSN: 1098-1004
Titre abrégé: Hum Mutat
Pays: United States
ID NLM: 9215429

Informations de publication

Date de publication:
01 2021
Historique:
received: 21 05 2020
revised: 09 10 2020
accepted: 24 11 2020
pubmed: 1 12 2020
medline: 1 4 2022
entrez: 30 11 2020
Statut: ppublish

Résumé

In genetic diseases, the most prevalent mechanism of pathogenicity is an altered expression of dosage-sensitive genes. Drugs that restore physiological levels of these genes should be effective in treating the associated conditions. We developed a screening strategy, based on a bicistronic dual-reporter vector, for identifying compounds that modulate protein levels, and used it in a pharmacological screening approach. To provide a proof-of-principle, we chose autosomal dominant leukodystrophy (ADLD), an ultra-rare adult-onset neurodegenerative disorder caused by lamin B1 (LMNB1) overexpression. We used a stable Chinese hamster ovary (CHO) cell line that simultaneously expresses an AcGFP reporter fused to LMNB1 and a Ds-Red normalizer. Using high-content imaging analysis, we screened a library of 717 biologically active compounds and approved drugs, and identified alvespimycin, an HSP90 inhibitor, as a positive hit. We confirmed that alvespimycin can reduce LMNB1 levels by 30%-80% in five different cell lines (fibroblasts, NIH3T3, CHO, COS-7, and rat primary glial cells). In ADLD fibroblasts, alvespimycin reduced cytoplasmic LMNB1 by about 50%. We propose this approach for effectively identifying potential drugs for treating genetic diseases associated with deletions/duplications and paving the way toward Phase II clinical trials.

Identifiants

pubmed: 33252173
doi: 10.1002/humu.24147
doi:

Substances chimiques

Lamin Type B 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

102-116

Informations de copyright

© 2020 Wiley Periodicals LLC.

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Auteurs

Elisa Giorgio (E)

Department of Medical Sciences, Medical Genetics Unit, University of Torino, Turin, Italy.
Department of Molecular Medicine, University of Pavia, Pavia, Italy.

Emanuela Pesce (E)

UOC Genetica Medica, IRCCS Istituto Giannina Gaslini, Genova, Italy.

Elisa Pozzi (E)

Department of Medical Sciences, Medical Genetics Unit, University of Torino, Turin, Italy.

Elvira Sondo (E)

UOC Genetica Medica, IRCCS Istituto Giannina Gaslini, Genova, Italy.

Marta Ferrero (M)

Department of Medical Sciences, Medical Genetics Unit, University of Torino, Turin, Italy.

Cristina Morerio (C)

UOC Laboratorio di Genetica Umana, IRCCS Istituto Giannina Gaslini, Genova, Italy.

Giusy Borrelli (G)

Department of Medical Sciences, Medical Genetics Unit, University of Torino, Turin, Italy.

Edoardo Della Sala (E)

Department of Medical Sciences, Medical Genetics Unit, University of Torino, Turin, Italy.

Martina Lorenzati (M)

Department of Neuroscience Rita Levi Montalcini and Neuroscience Institute Cavalieri Ottolenghi, University of Torino, Orbassano, Torino, Italy.

Pietro Cortelli (P)

Department of Biomedical and Neuromotor Sciences, University of Bologna, Bologna, Italy.
IRCCS Istituto delle Scienze Neurologiche di Bologna, Bellaria Hospital, Bologna, Italy.

Annalisa Buffo (A)

Department of Neuroscience Rita Levi Montalcini and Neuroscience Institute Cavalieri Ottolenghi, University of Torino, Orbassano, Torino, Italy.

Nicoletta Pedemonte (N)

UOC Genetica Medica, IRCCS Istituto Giannina Gaslini, Genova, Italy.

Alfredo Brusco (A)

Department of Medical Sciences, Medical Genetics Unit, University of Torino, Turin, Italy.
Medical Genetics Unit, Città della Salute e della Scienza University Hospital, Turin, Italy.

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