Dados do Trabalho

Título

Development of a 3D co-culture model for lung cancer lines and lymphocytes under anti-PD-1 and anti-PD-L1 treatment

Introdução

Despite the great therapeutic advances brought by immune checkpoint inhibitors (anti-CTLA-4 and anti-PD-1/PD-L1), the response rate for non-small cell lung cancer (NSCLC) is only 20%. The development of methodologies to study the interaction between the immune system and the tumor microenvironment allow the study of the interaction between cell populations, under conditions similar to the physiological one, being, a great promise for studies involving the combination of different therapies.

Objetivo

To establish 3D co-culture models involving NSCLC cell lines and peripheral blood lymphocytes from healthy individuals and to evaluate their interaction when treated with immunological checkpoint inhibitors.

Métodos

Initially, a comprehensive screening will be carried out to access PD-L1 expression in different NSCLC cell lines through Western Blotting technique. This assessment aims to identify positive, negative and intermediate expression for PD-L1. For the establishment of the co-culture model, cell lines will be co-cultured alongside CD8+ T lymphocytes, which will be isolated and activated through magnetic beads. Subsequent to co-culturing, these cells will be subjected to anti-PD1 and anti-PD-L1 treatment in order to evaluate their behavior and responses towards these inhibitors. To achieve this, cell viability (MTS), metabolic profile, immunophenotypic profile and immunological mediators (flow cytometry) were evaluated.

Resultados

Our findings revealed distinct PD-L1 expression patterns across the examined cell lines. A549 and H292 WT cell lines exhibited negligible PD-L1 expression, while H358 displayed significantly elevated levels. Meanwhile, H1975, HCC 827, SKMES1 and LCOR105 cell lines demonstrated intermediate PD-L1 expression. In terms of spheroid formation, H292 WT, A549, H358, HCC 827 and SKMES1 cell lines exhibited successful spheroid formation across all tested concentrations. However, spheroid formation was not observed in H1975 and LCOR105 cell lines. Notably, co-culture models involving H292 WT demonstrated an enhanced lymphocyte-tumor cell ratio of 3:1, underscoring its potential for facilitating improved interactions between these cell types.

Conclusões

The utilization of co-culture models merging tumor cell lines and lymphocytes emerges as a promising avenue with substantial potential for expediting drug screening and novel drug development endeavors.

Palavras-chave

Immunotherapy, co-culture, response to checkpoint inhibitors.

Financiador do resumo

This research was funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Brazil, grant number 2019/07111-9 and by institutional funding of Barretos Cancer Hospital (Brazil). LIMA, J.A. was a recipient of a scholarship from Barretos Cancer Hospital. Arantes, LMRB. was a recipient of a scholarship from FAPESP (2021/04100-6) and a recipient of a Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Produc-tivity (Brazil) fellowship.