The blood system serves as a key model for cell differentiation and cancer. It is orchestrated by precise spatiotemporal expression of crucial transcription factors. One of the key master regulators in the hematopoietic systems is PU.1. Reduced levels of PU.1 are characteristic for human acute myeloid leukemia (AML) and are known to induce AML in mouse models. Here, we demonstrate that transcriptional downregulation of PU.1 is an active process involving an alternative promoter in intron 3 that is induced by RUNX transcription factors driving non-coding antisense transcription. Core binding factor (CBF) fusions, RUNX1-ETO and CBFβ-MYH11 in t(8;21) and inv(16) AML, respectively, activate the PU.1 antisense promoter that results in a shift from sense towards antisense transcription and myeloid differentiation blockade. In patients with CBF-AML, we found that an elevated antisense/sense transcript and promoter accessibility ratio represents a hallmark compared to normal karyotype AML or healthy CD34+ cells. Competitive interaction of an enhancer with the proximal or the antisense promoter forms a binary on/off switch for either myeloid or T-cell development. Leukemic CBF fusions thus utilize a physiologic mechanism employed by T-cells to decrease sense transcription. Our study is the first example of a sense/antisense promoter competition as a crucial functional switch for gene expression perturbation by oncogenes. Hence, this disease mechanism reveals a previously unknown Achilles heel for future precise therapeutic targeting of oncogene-induced chromatin remodeling.
Emiel van der Kouwe, Gerwin Heller, Akos Czibere, John A Pulikkan, Christiane Agreiter, Lucio H Castilla, Ruud Delwel, Annalisa Di Ruscio, Alexander K Ebralidze, Maurizio Forte, Florian Grebien, Elizabeth Heyes, Lukas Kazianka, Julian Klinger, Christoph Kornauth, Trang Le, Karin Lind, Inês Amorim Monteiro Barbosa, Tea Pemovska, Alexander Sebastian Pichler, Ann-Sofie Schmolke, Carmen Marita Schweicker, Heinz Sill, Wolfgang R Sperr, Andreas Spittler, Sridevi Surapally, Bon Q Trinh, Peter Valent, Katrina Vanura, Robert S Welner, Johannes Zuber, Daniel G Tenen, Philipp B Staber