PT  - JOURNAL ARTICLE
AU  - Kristopher S. Raghavan
AU  - Ralph Francescone
AU  - Janusz Franco-Barraza
AU  - Jaye C. Gardiner
AU  - Débora Barbosa Vendramini-Costa
AU  - Tiffany Luong
AU  - Narges Pourmandi
AU  - Anthony Andren
AU  - Alison Kurimchak
AU  - Charline Ogier
AU  - James S. Duncan
AU  - Costas A. Lyssiotis
AU  - Lucia R. Languino
AU  - Edna Cukierman
TI  - NetrinG1<sup>+</sup> cancer-associated fibroblasts generate unique extracellular vesicles that support the survival of pancreatic cancer cells under nutritional stress
AID  - 10.1101/2021.11.21.469456
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.11.21.469456
4099  - http://biorxiv.org/content/early/2021/11/24/2021.11.21.469456.short
4100  - http://biorxiv.org/content/early/2021/11/24/2021.11.21.469456.full
AB  - It is projected that, in 5 years, pancreatic cancer will become the second deadliest cancer in the United States. A unique aspect of pancreatic ductal adenocarcinoma (PDAC) is its stroma; rich in cancer-associated fibroblasts (CAFs) and a dense CAF-generated extracellular matrix (ECM). This fibrous stroma, known as desmoplasia, causes the collapse of local blood vessels rendering a nutrient-deprived milieu. Hence, PDAC cells are nurtured by local CAF-secreted products, which include, among others, CAF-generated small extracellular vesicles (sEVs). It is well-accepted that upon culturing functionally tumor-promoting CAFs under pathophysiological-relevant conditions (e.g., within self-produced ECM), these cells express NetrinG1 (NetG1) and sustain endosomal pools rich in active α5β1-integrin; traits indicative of poor patient survival. We herein report that NetG1+ CAFs generate sEVs that rescue PDAC cells from nutrient-deprived induced apoptosis. Two unique sEVs, NetG1+ and α5β1-integrin+, were uncovered. The former constitutes cargo of CAF-generated exomeres, and the latter is detected in classic exosomes. Proteomic and metabolomic analyses showed that the sEV-dependent PDAC survival is, at least in part, dictated by the cargo packaged within sEVs in a NetG1-dependent manner. Indeed, despite producing a similar number of vesicles, selected key proteins and metabolites (e.g., glutamine) were incorporated within the unique sEVs. Finally, we found that NetG1 and α5β1-integrin were detected in sEVs collected from plasma of PDAC patients, while their concomitant levels were significantly lower in plasma of sex/age-matched healthy donors. The discovery of these tumor-supporting CAF sEVs opens a new investigative avenue in tumor-stroma interactions and stroma staging detection.Competing Interest StatementThe authors have declared no competing interest.