SULFATED HYALURONAN: AN IMMUNE MODULATOR OF THE TUMOR MICROENVIRONMENT?

SPINELLI, FIORELLA; VITALE, DAIANA; GUARISE, CRISTIAN; ICARDI, ANTONELA; DEL DAGO, DAIANA; PLUDA, STEFANO; GALESSO, DEVIS; ALANIZ, LAURA

Cleveland, Ohio

Conferencia; Conference on Advances in Basic, Applied and Clinical Science of Hyaluronan; 2017

International Society for Hyaluronan Sciences (ISHAS)

Introduction. Non-Small Cell Lung Cancer (NSCLC) is the most common type of lung cancer, having a mid-low rate of survival in different stages. Since there is no single best treatment, it is necessary to find a successful therapeutically option that complement the current protocols. Chemically modified hyaluronan, such as Sulfated hyaluronan (sHA), are a possible adjuvant therapy due to their proved action to: (i) inhibit HAses1, (ii) have antitumor and antiangiogenic activity in prostate and bladder cancer cells2, 3 and (iii) modulate endothelial cells behavior, blocking the action of VEGF4. However, direct effect in immune cells was not elucidated yet. Further investigations are needed to understand the immune and angiogenic signals triggered by sHA in the context of the tumor microenvironment. Aims. Evaluate the antitumoral action of sulfated hyaluronan with two different sulfation degrees (sHA1 and sHA3) on NSCLC cell line as well as the phenotype polarization of macrophages derived from PBMCs in the tumor context.Materials and methods. sHA were synthesized from the tetrabutyl ammonium salt of HA in Fidia Farmaceutici S.p.A. (Abano Terme, PD, Italy) (sHA1 MW=28 kDa and sHA3 MW=66 kDa). Human NSCLC cell line (H1299) was treated with different concentrations (0, 20, 100 and 1000 ug/ml) of sHA1 or sHA3. Macrophages derived from THP-1 cell line and human PBMCs, were treated with sHA3 with either H1299 tumor lysate or the conditioned media . Flow cytometry was used to measure apoptosis (Annexin V-APC) and to characterize macrophages (HLA II, CD80, CD163, CD206). Trypan Blue and MTT Assay were performed to evaluate cell viability. VEGF and TNF-α biosynthesis was measured by ELISA assay. Results. The treatment with sHA3 1000 ug/ml increased apoptosis levels in H1299, this effect was not showed with sHA1 in the same concentration. The cell viability of PBMCs derived macrophages wasn?t affected by sHA3 treatments with or without H1299 tumor lysates (TL) or supernatant from tumor (conditioned media: CM). Even though the expression of M1 and M2 cell surface markers (HLA, CD80, CD206) was not affected by sHA3 treatments in the tumor context, VEGF biosynthesis of macrophages diminished significantly in the presence of sHA3 1000 ug/ml plus TL but not of TL alone (724,4±221,4 and 1766±46,75 pg/mL respectively). Preliminary results show an increase in proinflammatory TNF synthesis in the presence of sHA3 in a dose- dependent manner. Discussion. A direct antitumor activity by sHA3 was observed due to apoptosis induction in NSCLC tumor cells. Even more, sHA3 could modulate immune activation by affecting the behavior of macrophages, reducing their angiogenic capability and enhancing their pro-inflammatory action. Funding Sources. GLYCAN 645756, European Commission. CONICET. Instituto Nacional del Cancer. References. 1 T. Isoyama, et al., Glycobiology, 2006, 16, 11-21. 2 A. Benitez, et al., Cancer research, 2011, 71, 4085-4095. 3 A. R. Jordan, et al., Oncotarget, 2016. 4 D. K. Lim, et al., Biomaterials, 2016, 77, 130-138.