Mimicking Human EED Variants in Drosophila: A Promising Strategy to Analyse Human EED Variant Function

AbstractThe Polycomb Repressive Complex 2 is an epigenetic reader/writer that methylates histone H3K27. Rare germline partial loss of function (pLoF) variants in core members of the complex (EZH2, EED, SUZ12) cause overgrowth and intellectual disability syndromes, whereas somatic variants are implicated in cancer. However, up to 1% of the general population will have a rare variant in one of these genes, most of which would be classified as variants of uncertain significance (VoUS). Towards screening these VoUS for partial LoF alleles that may contribute to disease, here we report functional assays inDrosophilato interrogateEEDmissense variants. We mimicked the amino acid change(s) ofEEDvariants into itsDrosophilaortholog,esc, and tested their function. Known likely benign variants functioned wildtype and known pathogenic variants were LoF. We further demonstrate the utility of this calibrated assay as a scalable approach to assist clinical interpretation of humanEEDVoUS.