Genes and compounds that increase type VII collagen expression as potential treatments for dystrophic epidermolysis bullosa

Research output: Contribution to journalArticlepeer-review

Abstract

Dystrophic epidermolysis bullosa (DEB) is a skin-blistering disease caused by mutations in COL7A1, which encodes type VII collagen (C7). There is no cure for DEB, but previous work has shown potential therapeutic benefit of increased production of even partially functional C7. Genome-wide screens using CRISPR-Cas9 have enabled the identification of genes involved in cancer development, drug resistance and other genetic diseases, suggesting that they could be used to identify drivers of C7 production. A keratinocyte C7 reporter cell line was created and used in a genome-wide CRISPR activation (CRISPRa) screen to identify genes and pathways that increase C7 expression. The CRISPRa screen results were used to develop a targeted drug screen to identify compounds that upregulate C7 expression. The C7_tdTomato cell line was validated as an effective reporter for detection of C7 upregulation. The CRISPRa screen identified DENND4B and TYROBP as top gene hits plus pathways related to calcium uptake and immune signalling in C7 regulation. The targeted drug screen identified several compounds that increase C7 expression in keratinocytes, of which kaempferol, a plant flavonoid, also significantly increased C7 mRNA and protein in DEB patient cells.

Original languageEnglish (US)
Pages (from-to)1065-1075
Number of pages11
JournalExperimental Dermatology
Volume31
Issue number7
DOIs
StatePublished - Jul 2022

Bibliographical note

Funding Information:
We thank our colleagues in the Tolar laboratory for their review and feedback on the draft manuscript. We thank the University of Minnesota Flow Cytometry Resource centre for their help with FACS. Dr. Thompson is partly funded by NIA Training Grant T32-AG029796 and the Children’s Cancer Research Fund Emerging Scientist Award. Drs. Tolar and Albert are partly supported by NIH grant R21-HG010380. Dr. Tolar is partly supported by NIH/NIAMS grant R01-AR063070. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Funding Information:
We thank our colleagues in the Tolar laboratory for their review and feedback on the draft manuscript. We thank the University of Minnesota Flow Cytometry Resource centre for their help with FACS. Dr. Thompson is partly funded by NIA Training Grant T32‐AG029796 and the Children’s Cancer Research Fund Emerging Scientist Award. Drs. Tolar and Albert are partly supported by NIH grant R21‐HG010380. Dr. Tolar is partly supported by NIH/NIAMS grant R01‐AR063070. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Publisher Copyright:
© 2022 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keywords

  • CRISPR-cas systems
  • kaempferol
  • keratinocytes
  • precision medicine
  • rare diseases

PubMed: MeSH publication types

  • Journal Article

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