A novel in vitro spheroid model enables multi-modality assessment of the anti-tumoral immune response in pancreatic neuroendocrine neoplasms

#3471

Introduction: Patients harbouring pancreatic neuroendocrine neoplasms (PNENs) often present as a non-resectable disease due to distant metastases, hence, efficacy of the current medical intervention is limited. Activation of the mammalian target of rapamycin (mTOR) pathway is a known tumorigenic driver in PNENs. mTOR inhibition by Everolimus (EVE) delays PNEN progression, but the mechanism is not clear. EVE is also used as an immunosuppressant, thus we hypothesized that it may suppress the anti-tumoral immune response.

Aim(s): To develop an in vitro model of PNEN that will imitate the anti-tumoral immune response and allow assessment such response with isolated treatment with mTOR inhibitors.

Materials and methods: BON-1 and MRC-5 spheroids were developed and were co-incubated with leukocytes from healthy donors. Leukocytes invasion to the spheroids were assessed by immunofluorescence (IF), using CD45 and CD3 antibodies. Synaptophysin and vimentin stainings were used to identify BON-1 and MRC-5, respectively. Nucleic acids were stained by 4',6-diamidino-2-phenylindole‎. The effect of the interventions on the spheroids' viability was assessed using CellTiter-Glo 3D Cell Viability Assay.

Conference:

Presenting Author:

Authors: Nasirov S, Zaig E, Mor-Cohen R, Telerman A, Tirosh A,

Keywords: PNEN, spheroid, Everolimus, mTOR,