Poster Presentation 35th Lorne Cancer Conference 2023

Systematic loss of function screens identify SMAD2 dervied circRNA as a target for WNT/β-catenin dependent cancers (#262)

Matthew Neve 1 , Ling Liu 1 , Laura Perlaza-Jimenez 1 , Azelle Hawdon 1 , Simon Conn 2 , Jennifer Zenker 1 , Pablo Tomayo 3 , Gregory Goodall 4 , Joseph Rosenbluh 1
  1. Monash University, Melbourne, Victoria, Australia
  2. Flinders University, Adelaide, South Australia, Australiua
  3. University of California, San Diego, San Diego, California
  4. Centre for Cancer Biology SA Pathology and University of South Australia, Adelaide, South Australia

Circular RNAs (circRNAs) are covalently closed single stranded RNAs that are produced by RNA back-splicing. Recent profiling demonstrates that circRNA expression is cell-type specific and dysregulated in many cancers. Of the thousands of known circRNAs, only a small number have been implicated as functional. This makes circRNA a functional engima and we lack a systematic understanding of the cancer associated signaling pathways that are regulated by circRNAs. Here, we generated a pooled shRNA lentiviral library targeting the back-splice junction of 3,354 human circRNAs that are expressed at low to high levels. We used this library for loss of function proliferation screens in a panel of 18 cancer cell lines from four tissue types (colon, pancreas, skin and brain) that harbour mutations leading to constitutive activity of defined pathways (Wnt/beta-catenin or MAPK). Using this dataset, we identify pathway specific and common-essential functional circRNAs using two-class comparisons of signaling pathway active and inactive cell lines. We validated these observations with a secondary screen and uncovered a role for circSMAD2, a novel regulator of the WNT/beta-catenin pathway. Our screening and validation assays demonstrate that circSMAD2 suppression leads to reduced proliferation and increased cell death only in cancer cells that are dependent on Wnt/beta-catenin signaling. Additionally, we show that circSMAD2 knockdown results in reduced beta-catenin protein stability. Our work adds a circular dimension to the regulation of cancer signalling pathways and identifies circSMAD2 as novel therapeutic target for Wnt/beta-catenin dependent cancers.