An NFATc1/SMAD3/cJUN Complex Restricted to SMAD4-Deficient Pancreatic Cancer Guides Rational Therapies

Marie C Hasselluhn; Denise Schlösser; Lennart Versemann; Geske E Schmidt; Maria Ulisse; Joana Oschwald; Zhe Zhang; Feda Hamdan; Harry Xiao; Waltraut Kopp; Jessica Spitalieri; Christin Kellner; Carolin Schneider; Kristina Reutlinger; Sankari Nagarajan; Benjamin Steuber; Stephen A Sastra; Carmine F Palermo; Jennifer Appelhans; Hanibal Bohnenberger; Jovan Todorovic; Irina Kostyuchek; Philipp Ströbel; Aiko Bockelmann; Alexander König; Christoph Ammer-Herrmenau; Laura Schmidleitner; Silke Kaulfuß; Bernd Wollnik; Stephan A Hahn; Albrecht Neesse; Shiv K Singh; Holger Bastians; Maximilian Reichert; Ulrich Sax; Kenneth P Olive; Steven A Johnsen; Günter Schneider; Volker Ellenrieder; Elisabeth Hessmann

doi:10.1053/j.gastro.2023.10.026

An NFATc1/SMAD3/cJUN Complex Restricted to SMAD4-Deficient Pancreatic Cancer Guides Rational Therapies

Gastroenterology. 2024 Feb;166(2):298-312.e14. doi: 10.1053/j.gastro.2023.10.026. Epub 2023 Oct 31.

Authors

Marie C Hasselluhn¹, Denise Schlösser², Lennart Versemann², Geske E Schmidt³, Maria Ulisse³, Joana Oschwald³, Zhe Zhang³, Feda Hamdan⁴, Harry Xiao⁵, Waltraut Kopp³, Jessica Spitalieri³, Christin Kellner³, Carolin Schneider⁶, Kristina Reutlinger², Sankari Nagarajan⁷, Benjamin Steuber², Stephen A Sastra⁵, Carmine F Palermo⁵, Jennifer Appelhans⁸, Hanibal Bohnenberger⁹, Jovan Todorovic⁸, Irina Kostyuchek⁹, Philipp Ströbel⁸, Aiko Bockelmann², Alexander König², Christoph Ammer-Herrmenau³, Laura Schmidleitner¹⁰, Silke Kaulfuß¹¹, Bernd Wollnik¹², Stephan A Hahn¹³, Albrecht Neesse³, Shiv K Singh³, Holger Bastians¹⁴, Maximilian Reichert¹⁵, Ulrich Sax¹⁶, Kenneth P Olive⁵, Steven A Johnsen¹⁷, Günter Schneider¹⁸, Volker Ellenrieder¹⁹, Elisabeth Hessmann²⁰

Affiliations

¹ Department of Gastroenterology, Gastrointestinal Oncology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany; Department of Medicine, Division of Digestive and Liver Diseases, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, New York; Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York.
² Department of Gastroenterology, Gastrointestinal Oncology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany.
³ Department of Gastroenterology, Gastrointestinal Oncology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany; Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany.
⁴ Gene Regulatory Mechanisms and Molecular Epigenetics Laboratory, Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, Minnesota.
⁵ Department of Medicine, Division of Digestive and Liver Diseases, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, New York; Herbert Irving Comprehensive Cancer Center, Columbia University Irving Medical Center, New York, New York.
⁶ Department of General, Visceral and Pediatric Surgery, University Medical Center Goettingen, Goettingen, Germany.
⁷ Manchester Breast Centre and Manchester Cancer Research Centre, Division of Molecular and Cellular Function, School of Biological Sciences, Faculty of Biology, Medicine and Health, The University of Manchester, Manchester, United Kingdom.
⁸ Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany; Institute of Pathology, University Medical Center Goettingen, Goettingen, Germany.
⁹ Institute of Pathology, University Medical Center Goettingen, Goettingen, Germany.
¹⁰ Medical Clinic and Polyclinic II, Klinikum Rechts der Isar, Technical University Munich, Munich, Germany; Translational Pancreatic Research Cancer Center, Medical Clinic and Polyclinic II, Klinikum Rechts der Isar, Technical University Munich, Munich, Germany.
¹¹ Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany; Institute of Human Genetics, University Medical Center Goettingen, Goettingen, Germany.
¹² Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany; Institute of Human Genetics, University Medical Center Goettingen, Goettingen, Germany; Cluster of Excellence Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells, University of Goettingen, Germany.
¹³ Ruhr University Bochum, Faculty of Medicine, Department of Molecular Gastrointestinal Oncology, Bochum, Germany.
¹⁴ Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany; Department of Molecular Oncology, Section for Cellular Oncology, University Medical Center Goettingen, Goettingen, Germany.
¹⁵ Medical Clinic and Polyclinic II, Klinikum Rechts der Isar, Technical University Munich, Munich, Germany; Translational Pancreatic Research Cancer Center, Medical Clinic and Polyclinic II, Klinikum Rechts der Isar, Technical University Munich, Munich, Germany; German Cancer Consortium (a partnership between Deutsches Krebsforschungszentrum and University Hospital Klinikum Rechts der Isar), Munich, Germany; Center for Protein Assemblies, Technical University of Munich, Garching, Germany; Center for Organoid Systems and Tissue Engineering, Technical University Munich, Garching, Germany.
¹⁶ Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany; Department of Medical Informatics, University Medical Center Goettingen, Goettingen, Germany.
¹⁷ Department of General, Visceral and Pediatric Surgery, University Medical Center Goettingen, Goettingen, Germany; Robert Bosch Center for Tumor Diseases, Stuttgart, Germany.
¹⁸ Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany; Department of General, Visceral and Pediatric Surgery, University Medical Center Goettingen, Goettingen, Germany; Comprehensive Cancer Center, Lower Saxony, Goettingen and Hannover, Germany.
¹⁹ Department of Gastroenterology, Gastrointestinal Oncology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany; Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany; Comprehensive Cancer Center, Lower Saxony, Goettingen and Hannover, Germany.
²⁰ Department of Gastroenterology, Gastrointestinal Oncology and Endocrinology, University Medical Center Goettingen, Goettingen, Germany; Clinical Research Unit KFO5002, University Medical Center Goettingen, Goettingen, Germany; Comprehensive Cancer Center, Lower Saxony, Goettingen and Hannover, Germany. Electronic address: [email protected].

PMID: 37913894
DOI: 10.1053/j.gastro.2023.10.026

Abstract

Background & aims: The highly heterogeneous cellular and molecular makeup of pancreatic ductal adenocarcinoma (PDAC) not only fosters exceptionally aggressive tumor biology, but contradicts the current concept of one-size-fits-all therapeutic strategies to combat PDAC. Therefore, we aimed to exploit the tumor biological implication and therapeutic vulnerabilities of a clinically relevant molecular PDAC subgroup characterized by SMAD4 deficiency and high expression of the nuclear factor of activated T cells (SMAD4^-/-/NFATc1^High).

Methods: Transcriptomic and clinical data were analyzed to determine the prognostic relevance of SMAD4^-/-/NFATc1^High cancers. In vitro and in vivo oncogenic transcription factor complex formation was studied by immunoprecipitation, proximity ligation assays, and validated cross model and species. The impact of SMAD4 status on therapeutically targeting canonical KRAS signaling was mechanistically deciphered and corroborated by genome-wide gene expression analysis and genetic perturbation experiments, respectively. Validation of a novel tailored therapeutic option was conducted in patient-derived organoids and cells and transgenic as well as orthotopic PDAC models.

Results: Our findings determined the tumor biology of an aggressive and chemotherapy-resistant SMAD4^-/-/NFATc1^High subgroup. Mechanistically, we identify SMAD4 deficiency as a molecular prerequisite for the formation of an oncogenic NFATc1/SMAD3/cJUN transcription factor complex, which drives the expression of RRM1/2. RRM1/2 replenishes nucleoside pools that directly compete with metabolized gemcitabine for DNA strand incorporation. Disassembly of the NFATc1/SMAD3/cJUN complex by mitogen-activated protein kinase signaling inhibition normalizes RRM1/2 expression and synergizes with gemcitabine treatment in vivo to reduce the proliferative index.

Conclusions: Our results suggest that PDAC characterized by SMAD4 deficiency and oncogenic NFATc1/SMAD3/cJUN complex formation exposes sensitivity to a mitogen-activated protein kinase signaling inhibition and gemcitabine combination therapy.

Keywords: MEK inhibition; NFATc1; Pancreatic Cancer; RRM1/2; Stratification.

MeSH terms

Carcinoma, Pancreatic Ductal* / drug therapy
Carcinoma, Pancreatic Ductal* / genetics
Carcinoma, Pancreatic Ductal* / metabolism
Cell Line, Tumor
Gemcitabine
Humans
Mitogen-Activated Protein Kinases / metabolism
Pancreatic Neoplasms* / drug therapy
Pancreatic Neoplasms* / genetics
Pancreatic Neoplasms* / metabolism
Smad3 Protein / metabolism
Smad4 Protein / genetics
Smad4 Protein / metabolism

Substances

Gemcitabine
Smad4 Protein
Mitogen-Activated Protein Kinases
SMAD4 protein, human
SMAD3 protein, human
Smad3 Protein