Cancer-associated fibroblast-derived annexin A6+ extracellular vesicles support pancreatic cancer aggressiveness
Julie Leca, Sébastien Martinez, Sophie Lac, Jérémy Nigri, Véronique Secq, Marion Rubis, Christian Bressy, Arnauld Sergé, Marie-Noelle Lavaut, Nelson Dusetti, Céline Loncle, Julie Roques, Daniel Pietrasz, Corinne Bousquet, Stéphane Garcia, Samuel Granjeaud, Mehdi Ouaissi, Jean Baptiste Bachet, Christine Brun, Juan L. Iovanna, Pascale Zimmermann, Sophie Vasseur, Richard Tomasini
Julie Leca, Sébastien Martinez, Sophie Lac, Jérémy Nigri, Véronique Secq, Marion Rubis, Christian Bressy, Arnauld Sergé, Marie-Noelle Lavaut, Nelson Dusetti, Céline Loncle, Julie Roques, Daniel Pietrasz, Corinne Bousquet, Stéphane Garcia, Samuel Granjeaud, Mehdi Ouaissi, Jean Baptiste Bachet, Christine Brun, Juan L. Iovanna, Pascale Zimmermann, Sophie Vasseur, Richard Tomasini
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Research Article Cell biology Oncology

Cancer-associated fibroblast-derived annexin A6+ extracellular vesicles support pancreatic cancer aggressiveness

Abstract

The intratumoral microenvironment, or stroma, is of major importance in the pathobiology of pancreatic ductal adenocarcinoma (PDA), and specific conditions in the stroma may promote increased cancer aggressiveness. We hypothesized that this heterogeneous and evolving compartment drastically influences tumor cell abilities, which in turn influences PDA aggressiveness through crosstalk that is mediated by extracellular vesicles (EVs). Here, we have analyzed the PDA proteomic stromal signature and identified a contribution of the annexin A6/LDL receptor-related protein 1/thrombospondin 1 (ANXA6/LRP1/TSP1) complex in tumor cell crosstalk. Formation of the ANXA6/LRP1/TSP1 complex was restricted to cancer-associated fibroblasts (CAFs) and required physiopathologic culture conditions that improved tumor cell survival and migration. Increased PDA aggressiveness was dependent on tumor cell–mediated uptake of CAF-derived ANXA6+ EVs carrying the ANXA6/LRP1/TSP1 complex. Depletion of ANXA6 in CAFs impaired complex formation and subsequently impaired PDA and metastasis occurrence, while injection of CAF-derived ANXA6+ EVs enhanced tumorigenesis. We found that the presence of ANXA6+ EVs in serum was restricted to PDA patients and represents a potential biomarker for PDA grade. These findings suggest that CAF–tumor cell crosstalk supported by ANXA6+ EVs is predictive of PDA aggressiveness, highlighting a therapeutic target and potential biomarker for PDA.

Authors

Julie Leca, Sébastien Martinez, Sophie Lac, Jérémy Nigri, Véronique Secq, Marion Rubis, Christian Bressy, Arnauld Sergé, Marie-Noelle Lavaut, Nelson Dusetti, Céline Loncle, Julie Roques, Daniel Pietrasz, Corinne Bousquet, Stéphane Garcia, Samuel Granjeaud, Mehdi Ouaissi, Jean Baptiste Bachet, Christine Brun, Juan L. Iovanna, Pascale Zimmermann, Sophie Vasseur, Richard Tomasini

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Figure 1

Human PDA microdissection followed by mass spectrometry coupled to bioinformatics analyses identifies a new complex in stroma PDA.

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Human PDA microdissection followed by mass spectrometry coupled to bioin...
(A) Number of proteins (and representative total percentage) obtained after mass spectrometry analysis of stroma and tumor cell areas microdissected from frozen human PDA slides (patients, n = 4; microdissected areas ranged from 30 to 50 mm2). (B) Graphical representation explaining how the complex comprising ANXA6, LRP1, and TSP1 was highlighted. (C) Twenty-two proteins belong to the same network of “cytoplasmic membrane-bound vesicles” obtained in Supplemental Figure 1D. Proteins in yellow boxes are specific to the stromal compartment, and proteins in yellow boxes surrounded by a red line are complex proteins obtained. (D) Western blot analysis of ANXA6, LRP1, and TSP1 in human healthy pancreas (#1–3) and PDA (#1–6). Amido black staining served as loading control. (E) Western blot analysis of ANXA6, LRP1, and TSP1 in murine healthy pancreas (#1–4) and PDA (#1–4). Amido black staining served as loading control. (F and G) Coimmunoprecipitation of LRP1 with ANXA6 and TSP1 in protein extracts from human (F) healthy pancreas (#2, #4) and PDA (#1, #7) and from murine (G) healthy pancreas (#5, #6) and PDA (#1, #4). Total cell lysate and nonrelevant antibody (NR) were used as loading and negative control, respectively. Data are representative of 3 independent experiments. Throughout the article, each “#” represents 1 PDA patient or mouse or 1 healthy donor or mouse.