ALANIZ Laura Daniela
congresos y reuniones científicas
SULFATED HYALURONAN: AN IMMUNE MODULATOR OF THE TUMOR MICROENVIRONMENT?
SPINELLI FIORELLA; VITALE DAIANA; GUARESI C; ICARDI A; DELDAGO D; PULDA S; GALLESO D; ALANIZ L
Conferencia; 11th International Conference Hyaluronan; 2017
International Society for Hyaluronan Sciences
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 shown 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 partially and fully sulfated hyaluronan (sHA1 and sHA3) in 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=28,1 kDa and sHA3 MW=66,4 kDa). Human NSCLC cell line (H1299) was treated with different concentrations (0, 20, 100 and 1000 ug/ml) of sHA1 o sHA3. Macrophages derived from THP-1 cell line and human PBMCs, were treated with sHA3 with or without 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 significant sHA3 1000 ug/ml plus TL in comparison to TL alone ( 1766,22). Preliminary results show an increase in proinflammatory TNF synthesis in dosis dependent manner of sHA3. Discussion. A direct antitumor action was observed by sHA3 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.