Extracellular Vesicles from Pancreatic Cancer Stem Cells Lead an Intratumor Communication Network (EVNet) to fuel tumour progression

Carolina F Ruivo; Nuno Bastos; Barbara Adem; Ines Batista; Cecilia Duraes; Carlos A Melo; Stephanie A Castaldo; Francisco Campos-Laborie; Pedro Moutinho-Ribeiro; Barbara Morão; Ana Costa-Pinto; Soraia Silva; Hugo Osorio; Sergio Ciordia; Jose Luis Costa; David Goodrich; Bruno Cavadas; Luisa Pereira; Tony Kouzarides; Guilherme Macedo; Rui Maio; Fatima Carneiro; Marília Cravo; Raghu Kalluri; Jose Carlos Machado; Sonia A Melo

doi:10.1136/gutjnl-2021-324994

Extracellular Vesicles from Pancreatic Cancer Stem Cells Lead an Intratumor Communication Network (EVNet) to fuel tumour progression

Gut. 2022 Jan 10;71(10):2043-2068. doi: 10.1136/gutjnl-2021-324994. Online ahead of print.

Authors

Carolina F Ruivo^#^{1

2

3}, Nuno Bastos^#^{1

2

3}, Barbara Adem^{1

2

3}, Ines Batista^{1

2

3}, Cecilia Duraes^{1

2}, Carlos A Melo⁴, Stephanie A Castaldo^{1

5}, Francisco Campos-Laborie⁴, Pedro Moutinho-Ribeiro^{6

7}, Barbara Morão⁸, Ana Costa-Pinto^{1

2}, Soraia Silva^{1

2}, Hugo Osorio^{1

2

6}, Sergio Ciordia⁹, Jose Luis Costa^{1

2

6}, David Goodrich¹⁰, Bruno Cavadas^{1

2}, Luisa Pereira^{1

2}, Tony Kouzarides⁴, Guilherme Macedo^{6

7}, Rui Maio^{8

11

12}, Fatima Carneiro^{1

2

6

7}, Marília Cravo^{11

13}, Raghu Kalluri¹⁴, Jose Carlos Machado^{1

2

6}, Sonia A Melo^{1

2

6}

Affiliations

¹ i3S Instituto de Investigação e Inovação em Saúde, University of Porto, Porto, Portugal.
² IPATIMUP Institute of Molecular Pathology and Immunology, University of Porto, Porto, Portugal.
³ ICBAS Instituto de Ciencias Biomédicas Abel Salazar, University of Porto, Porto, Portugal.
⁴ Gurdon Institute, Cambridge, Cambridgeshire, UK.
⁵ Department of Oncology, VIB-KU Leuven Center for Cancer Biology, Leuven, Belgium.
⁶ FMUP Faculty of Medicine University of Porto, Porto, Portugal.
⁷ CHUSJ Centro Hospitalar Universitário de São João, Porto, Portugal.
⁸ Hospital Beatriz Ângelo, Loures, Portugal.
⁹ Proteomics Facility, Spanish National Center for Biotechnology, Madrid, Spain.
¹⁰ Roswell Park Cancer Institute, Buffalo, New York, USA.
¹¹ Hospital da Luz, Lisbon, Portugal.
¹² NOVA Medical School, Lisbon, Portugal.
¹³ FMUL Faculty of Medicine University of Lisbon, Lisbon, Portugal.
¹⁴ Cancer Biology, University Texas MD Anderson Cancer Center, Houston, Texas, USA.

^# Contributed equally.

Abstract

Objective: Intratumor heterogeneity drives cancer progression and therapy resistance. However, it has yet to be determined whether and how subpopulations of cancer cells interact and how this interaction affects the tumour.

Design: We have studied the spontaneous flow of extracellular vesicles (EVs) between subpopulations of cancer cells: cancer stem cells (CSC) and non-stem cancer cells (NSCC). To determine the biological significance of the most frequent communication route, we used pancreatic ductal adenocarcinoma (PDAC) orthotopic models, patient-derived xenografts (PDXs) and genetically engineered mouse models (GEMMs).

Results: We demonstrate that PDAC tumours establish an organised communication network between subpopulations of cancer cells using EVs called the EVNet). The EVNet is plastic and reshapes in response to its environment. Communication within the EVNet occurs preferentially from CSC to NSCC. Inhibition of this communication route by impairing Rab27a function in orthotopic xenographs, GEMMs and PDXs is sufficient to hamper tumour growth and phenocopies the inhibition of communication in the whole tumour. Mechanistically, we provide evidence that CSC EVs use agrin protein to promote Yes1 associated transcriptional regulator (YAP) activation via LDL receptor related protein 4 (LRP-4). Ex vivo treatment of PDXs with antiagrin significantly impairs proliferation and decreases the levels of activated YAP.Patients with high levels of agrin and low inactive YAP show worse disease-free survival. In addition, patients with a higher number of circulating agrin⁺ EVs show a significant increased risk of disease progression.

Conclusion: PDAC tumours establish a cooperation network mediated by EVs that is led by CSC and agrin, which allows tumours to adapt and thrive. Targeting agrin could make targeted therapy possible for patients with PDAC and has a significant impact on CSC that feeds the tumour and is at the centre of therapy resistance.

Keywords: carcinogenesis; cell biology; molecular carcinogenesis; pancreatic cancer.

Abstract

Grants and funding