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EACR The Tumour Ecosystem: Cellular Interactions and Therapeutic Opportunities Conference, Bergamo, Italy, 19 - 21 March 2024, pp.1, (Summary Text)
Background: Non-small cell lung cancer (NSCLC) is diagnosed at an advanced stage due to rapid metastasis with poor prognosis requiring drug candidates targeting tumor microenvironment. Our group previously revealed antitumoral effect of cannabinoid 1 (CB1) agonist Arachidonoylcyclopropylamide (ACPA) on NSCLC through PI3K/Akt/JNK (PCT/TR2020/050618). Herein, we hypothesized that ACPA might have an antitumoral potential to reverse transformation of human normal lung fibroblasts (hNLF) to cancer-associated fibroblast (CAF) phenotype through NSCLC-derived-exosomes in vitro.
Methods: Effective dose of ACPA-administered A549-luc NSCLC-exosomes were isolated by ultracentrifugation, characterized by BCA for protein concentration, Zeta Sizer for particle size (PS), zeta potential (ZP), polydispersity index (PDI), flow cytometry (FCM) for CD9/CD63/CD81, and TEM for size. Real-time qPCR assessed miR-21/miR-23 expressions in control and ACPA-treated NSCLC-exosomes. MTT and ELISA assessed effectivity of ACPA-treated NSCLC cell-derived-exosomes on proliferative index and secretome profile of LL24 hNLFs.
Results: Control and ACPA-treated NSCLC-exosomes had a total protein concentration of 312 and 461 µg/ml, respectively. PS, ZP and PDI of control and ACPA-treated NSCLC-exosomes were 251 and 317 nm, -12 and -23 mV, and 0.498 and 0.563 respectively. CD9, CD63 and CD81 were detected as 96.93%, 98.35% and 77.33% in control exosomes, whereas, 95.75%, 95.86% and 70.16%, respectively, in ACPA-treated cell exosomes. The mean diameters of control and ACPA-treated NSCLC-exosomes were 147 nm and 132 nm. ACPA-treated NSCLC-exosomes presented lower miR-21 and miR-23 expressions, inhibited proliferation and IL-6/8 secretion of LL24 fibroblasts.
Conclusion: This study reveals the inhibition of transformation of hNLFs to CAF phenotype through ACPA-administered NSCLC-derived exosome delivery for the first time. Non-psychoactive low dose of ACPA might present a potential chemotherapeutic potency on CAF transformation through NSCLC-exosome release in tumor microenvironment.
(*) Hacettepe University Scientific Research Projects Coordination Unit (#TSA-2023-20427) and Scientific and Technological Research Council of Turkey (TUBITAK) (partially #122S459) supported this study.