GAPDH Controls Transcript Stability, Protein Translation and Drives Proliferation in Acute Myeloid Leukemia [GAPDH_KD_RNAseq]

Dysregulation of RNA binding proteins (RBPs) is a hallmark in cancerous cells. In acute myeloid leukemia (AML) RBPs are key regulators of tumor proliferation. While classical RBPs have defined RNA binding domains, RNA recognition and function in AML by non-canonical RBPs (ncRBPs) remains unclear. Given the inherent complexity of targeting AML broadly, our goal was to uncover potential ncRBP candidates critical for AML survival using a CRISPR/Cas-based screening. We identified the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a pro-proliferative factor in AML cells. Based on cross-linking and immunoprecipitation (CLIP) we are defining the global targetome detecting novel RNA targets mainly located within 5’UTR including GAPDH, RPL13a, PKM and ENO1. The knockdown of GAPDH unveiled genetic pathways related to ribosome biogenesis, translation initiation, and regulation. Moreover, we were able to demonstrate a stabilizing effect through the GAPDH binding to target transcripts including its own mRNA, unveiling GAPDH’s pivotal role in cancer. The present findings provide new insights on the pathophysiological role of GAPDH in AML.