Cancer promoting transcriptional enhancers controlled by the EMT-activator ZEB1

• Applicants: Professor Dr. Thomas Brabletz; Privatdozentin Dr. Simone Brabletz
• Subject Area: Pathology ; Hematology, Oncology
• Term: from 2019 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 416775465

• Final Report Year: 2023

Final Report Abstract

To date, the vast majority of cancer related deaths is caused by the formation of distant metastases. Thus, it is of utmost importance to better understand and predict the metastatic process. During the last two decades we and others could identify cancer cell plasticity, mediated by the embryonic program of Epithelial to Mesenchymal Transition (EMT) and its reverse process (MET), to be a crucial trait driving the metastatic cascade. Our work focuses on the transcription factor ZEB1, a major EMT inducer and known repressor of epithelial genes. We have shown that in aggressive cancers, ZEB1 can turn into a transcriptional activator of tumor-promoting genes. Lately, gene activation via distant transcriptional enhancers emerged as an important metastasis promoting mechanism. Active enhancers are even considered as prognostic and potentially diagnostic tumor markers. Within this project, we have shown that ZEB1, in cooperation with YAP and AP1, functions in enhancer activation. We characterized ZEB1 dependent enhancer regions on a genome wide level and annotated those enhancers to the respective regulated genes. One of the top regulated genes was NT5E (CD73), an immune dampening factor that is in focus as a potential target for novel immune modulating anti-tumour therapies. We could experimentally prove the ZEB1 dependent, enhancer mediated regulation of NT5E and found ZEB1 and NT5E expression to be strongly correlated in breast cancer patient samples with high expression indicating lower survival probability. We thus identified a novel aspect of ZEB1 dependent tumour promoting functions which also represents a targetable Achilles heel, since NT5E inhibiting antibodies are already available. Further, we now have a large list of ZEB1 dependent enhancers and corresponding genes that will be further explored and will be useful to identify novel pro-metastatic enhancers and genes that might serve as potential prognostic, diagnostic or therapeutic tools. In summary, we consider the project as very successful. Data of project was integrated in six publications in high ranked journals (five original and one review article). Over the years of funding we generated a huge fundus of data, which was the basis for a number of follow-up third party funded projects. These include projects in the newly granted SFB/TRR 305, a DFG-project granted for the SPP 2306, as well as a DFG granted Reinhart Kosseleck Project.

Publications

• Genome‐wide cooperation of EMT transcription factor ZEB 1 with YAP and AP ‐1 in breast cancer. The EMBO Journal, 39(17).
  Feldker, Nora; Ferrazzi, Fulvia; Schuhwerk, Harald; Widholz, Sebastian A.; Guenther, Kerstin; Frisch, Isabell; Jakob, Kathrin; Kleemann, Julia; Riegel, Dania; Bönisch, Ulrike; Lukassen, Sören; Eccles, Rebecca L.; Schmidl, Christian; Stemmler, Marc P.; Brabletz, Thomas & Brabletz, Simone
  
• Dynamic EMT: a multi‐tool for tumor progression. The EMBO Journal, 40(18).
  Brabletz, Simone; Schuhwerk, Harald; Brabletz, Thomas & Stemmler, Marc P.
  
• Pancreas morphogenesis and homeostasis depends on tightly regulated Zeb1 levels in epithelial cells. Cell Death Discovery, 7(1).
  Lasierra, Losada María; Pauler, Melissa; Vandamme, Niels; Goossens, Steven; Berx, Geert; Leppkes, Moritz; Schuhwerk, Harald; Brabletz, Simone; Brabletz, Thomas & Stemmler, Marc P.
  
• The EMT transcription factor ZEB1 blocks osteoblastic differentiation in bone development and osteosarcoma. The Journal of Pathology, 254(2), 199-211.
  Ruh, Manuel; Stemmler, Marc P.; Frisch, Isabell; Fuchs, Kathrin; van Roey, Ruthger; Kleemann, Julia; Roas, Maike; Schuhwerk, Harald; Eccles, Rebecca L.; Agaimy, Abbas; Baumhoer, Daniel; Berx, Geert; Müller, Fabian; Brabletz, Thomas & Brabletz, Simone
  
• The transcription factor ZEB1 regulates stem cell self-renewal and cell fate in the adult hippocampus. Cell Reports, 36(8), 109588.
  Gupta, Bhavana; Errington, Adam C.; Jimenez-Pascual, Ana; Eftychidis, Vasileios; Brabletz, Simone; Stemmler, Marc P.; Brabletz, Thomas; Petrik, David & Siebzehnrubl, Florian A.
  
• The EMT transcription factor ZEB1 governs a fitness-promoting but vulnerable DNA replication stress response. Cell Reports, 41(11), 111819.
  Schuhwerk, Harald; Kleemann, Julia; Gupta, Pooja; van Roey, Ruthger; Armstark, Isabell; Kreileder, Martina; Feldker, Nora; Ramesh, Vignesh; Hajjaj, Yussuf; Fuchs, Kathrin; Mahapatro, Mousumi; Hribersek, Mojca; Volante, Marco; Groenewoud, Arwin; Engel, Felix B.; Ceppi, Paolo; Eckstein, Markus; Hartmann, Arndt; Müller, Fabian ... & Brabletz, Thomas