Combination of treatment with hyaluronan plus doxorubicin on hematopoietic and solid tumor cells affects endothelial cell behavior independently of VEGF expression

DAIANA VITALE; ANTONELLA ICARDI; FIORELLA SPINELLI; INA SEVIC; GIANINA DEMARCHI; ILARIA CAON; DAVIDE VIGETTI; ALBERTO PASSI; CAROLINA CRISTINA; LAURA ALANIZ

Conferencia; 13th conference of the International Society for Hyaluronan Sciences; 2021

ISHAS

Hyaluronan (HA) is one of the main glycosaminoglycans of extracellular matrices and it is concentrated in tissues with high cell proliferation and migration rates (1). In several pathologies as cancer, HA expression is altered and it becomes fragmented into low-molecular-weight (LMW) forms, affecting mechanisms associated with cell proliferation, invasion, angiogenesis and multidrug resistance (2,3). We analyzed the effect of LMW HA on the response of T-lymphoma and mammary adenocarcinoma cell lines to the antineoplastic drug doxorubicin (DOX). Even more, we investigated if combination with HA and DOX treatment modifies the behavior of endothelial cells and consequently angiogenic response in both types of tumors. To alter tumor extracellular matrix, EL4 and MDA-MB-231 cells were first treated with LMW HA, and after 12 hours, DOX was added to perform a co-treatment during 12 hours. LMW HA altered DOX accumulation and modulated the expression of ATP-binding cassette transporters only in T-cell lymphoma cells in a conventional 2D culture model. In contrast, no changes in drug accumulation were observed in breast cancer cells. To further approximate the breast cancer model, we performed a 3D-culture strategy with LMW HA and DOX treatment. In that model, we observed a lower accumulation of DOX in the presence of LMW HA together with lower apoptosis rates, which demonstrated the role of HA in a 3D tumor context. When we evaluated the effect on angiogenic mechanisms, the supernatant from tumor cells treated with DOX exhibited a pro-angiogenic effect on endothelial cells. Combination of LMW HA + DOX increased migration and vessel formation in endothelial cells. This effect was independent of vascular endothelial growth factor but related to fibroblast growth factor-2 expression. Besides, we observed a pro-angiogenic effect on endothelial cells during combination of LMW HA + DOX treatment in the in vivo murine models of T-lymphoma and breast cancer, confirmed with higher migration of endothelial cells and vessel formation in tumor microenvironment. Our results demonstrated for the first time that HA is a potential modulator of DOX response involving drug efflux and angiogenic processes and consequently providing an adverse tumor stroma during chemotherapy.References: 1. F Spinelli et al ? Hyaluronan in the Tumor Microenvironment (2020) - Tumor Microenvironment - pp 67-83 - ISBN: 978-3-030-40146-72. T Chanmee et al- Cancer Letters (2016) 28, 375(1):20-30; doi: 10.1016/j.canlet.2016.02.031.3. A Passi et al - The FEBS journal (2019) 286, (15):2937-2949; doi: 10.1111/febs.14847.