CRISPR Activation Screening Identifies VGLL3-TEAD1-RUNX1/3 as a Transcriptional Complex for PD-L1 Expression

J Immunol. 2022 Sep 1;209(5):907-915. doi: 10.4049/jimmunol.2100917. Epub 2022 Aug 3.


The PD-L1/2-PD-1 immune checkpoint is essential for the proper induction of peripheral tolerance and limits autoimmunity, whereas tumor cells exploit their expression to promote immune evasion. Many different cell types express PD-L1/2, either constitutively or upon stimulation, but the factors driving this expression are often poorly defined. In this study, using genome-wide CRISPR activation screening, we identified three factors that upregulate PD-L1 expression: GATA2, MBD6, and transcription cofactor vestigial-like protein 3 (VGLL3). VGLL3 acts as a transcriptional regulator, and its expression induced PD-L1 in many different cell types. Conversely, loss of VGLL3 impaired IFN-γ-induced PD-L1/2 expression in human keratinocytes. Mechanistically, by performing a second screen to identify proteins acting in concert with VGLL3, we found that VGLL3 forms a complex with TEAD1 and RUNX1/3 to drive expression of PD-L1/2. Collectively, our work identified a new transcriptional complex controlling PD-L1/2 expression and suggests that VGLL3, in addition to its known role in the expression of proinflammatory genes, can balance inflammation by upregulating the anti-inflammatory factors PD-L1 and PD-L2.

MeSH terms

  • B7-H1 Antigen*
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics
  • Core Binding Factor Alpha 2 Subunit / genetics
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Humans
  • Immune Evasion
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Programmed Cell Death 1 Receptor* / genetics
  • TEA Domain Transcription Factors
  • Transcription Factors / genetics


  • B7-H1 Antigen
  • CD274 protein, human
  • Core Binding Factor Alpha 2 Subunit
  • DNA-Binding Proteins
  • Nuclear Proteins
  • Programmed Cell Death 1 Receptor
  • RUNX1 protein, human
  • TEA Domain Transcription Factors
  • TEAD1 protein, human
  • Transcription Factors
  • VGLL3 protein, human