ZFIN ID: ZDB-PUB-200403-190
Applying genome-wide CRISPR-Cas9 screens for therapeutic discovery in facioscapulohumeral muscular dystrophy
Lek, A., Zhang, Y., Woodman, K.G., Huang, S., DeSimone, A.M., Cohen, J., Ho, V., Conner, J., Mead, L., Kodani, A., Pakula, A., Sanjana, N., King, O.D., Jones, P.L., Wagner, K.R., Lek, M., Kunkel, L.M.
Date: 2020
Source: Science Translational Medicine   12(536): (Journal)
Registered Authors: Kunkel, Louis M., Lek, Angela
Keywords: none
MeSH Terms:
  • Animals
  • CRISPR-Cas Systems/genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics
  • Homeodomain Proteins/genetics
  • Homeodomain Proteins/metabolism
  • Humans
  • Muscular Dystrophy, Facioscapulohumeral*/genetics
  • Muscular Dystrophy, Facioscapulohumeral*/therapy
  • Zebrafish/genetics
  • Zebrafish/metabolism
PubMed: 32213627 Full text @ Sci. Transl. Med.
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ABSTRACT
The emergence of CRISPR-Cas9 gene-editing technologies and genome-wide CRISPR-Cas9 libraries enables efficient unbiased genetic screening that can accelerate the process of therapeutic discovery for genetic disorders. Here, we demonstrate the utility of a genome-wide CRISPR-Cas9 loss-of-function library to identify therapeutic targets for facioscapulohumeral muscular dystrophy (FSHD), a genetically complex type of muscular dystrophy for which there is currently no treatment. In FSHD, both genetic and epigenetic changes lead to misexpression of DUX4, the FSHD causal gene that encodes the highly cytotoxic DUX4 protein. We performed a genome-wide CRISPR-Cas9 screen to identify genes whose loss-of-function conferred survival when DUX4 was expressed in muscle cells. Genes emerging from our screen illuminated a pathogenic link to the cellular hypoxia response, which was revealed to be the main driver of DUX4-induced cell death. Application of hypoxia signaling inhibitors resulted in increased DUX4 protein turnover and subsequent reduction of the cellular hypoxia response and cell death. In addition, these compounds proved successful in reducing FSHD disease biomarkers in patient myogenic lines, as well as improving structural and functional properties in two zebrafish models of FSHD. Our genome-wide perturbation of pathways affecting DUX4 expression has provided insight into key drivers of DUX4-induced pathogenesis and has identified existing compounds with potential therapeutic benefit for FSHD. Our experimental approach presents an accelerated paradigm toward mechanistic understanding and therapeutic discovery of a complex genetic disease, which may be translatable to other diseases with well-established phenotypic selection assays.
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