A Macrophages-Enriched Head and Neck Tumor Spheroid Model to Study Foslip® Behavior in Tumor Microenvironment

Aurélie Francois; Luca Dirheimer; Alicia Chateau; Henri-Pierre Lassalle; Ilya Yakavets; Lina Bezdetnaya

doi:10.2147/IJN.S427350

A Macrophages-Enriched Head and Neck Tumor Spheroid Model to Study Foslip^® Behavior in Tumor Microenvironment

Int J Nanomedicine. 2023 Nov 9:18:6545-6562. doi: 10.2147/IJN.S427350. eCollection 2023.

Authors

Aurélie Francois^{1

2}, Luca Dirheimer^{1

2}, Alicia Chateau², Henri-Pierre Lassalle^{1

2}, Ilya Yakavets³, Lina Bezdetnaya^{1

2}

Affiliations

¹ Research Department, Institut de Cancérologie de Lorraine, Vandoeuvre-lès-Nancy, France.
² Centre de Recherche en Automatique de Nancy, Centre National de la Recherche Scientifique, UMR 7039, Université de Lorraine, Vandoeuvre-lès-Nancy, France.
³ Department of Chemistry, University of Toronto, Toronto, Canada.

Abstract

Purpose: The tumor microenvironment (TME) is composed of various stromal components, including immune cells such as tumor-associated macrophages (TAMs), which play a crucial role in cancer initiation and progression. TAMs can exhibit either a tumor-suppressive M1 or a tumor-promoting M2 phenotype. First, we aimed to develop a 3D human heterotypic model consisting of head and neck squamous cell carcinoma (HNSCC) cells and different subtypes of macrophages to replicate the interactions between immune cells and cancer cells. We further investigated the behavior of Foslip^®, a liposomal formulation of temoporfin, using a macrophage-enriched 3D model.

Methods: Monocytes were differentiated into M1 and M2 macrophages, which represent two distinct subtypes. Following histological and molecular characterization, these macrophages were used to establish a 3D spheroid model of HNSCC enriched with either polarized macrophages or conditioned media. Flow cytometry and fluorescence microscopy were used to assess the accumulation and distribution of Foslip^®. The cytotoxic effect of Foslip^®-mediated photodynamic therapy (PDT) was evaluated using flow cytometry.

Results: We developed heterotypic spheroids characterized by a mixed phenotype of evenly distributed macrophages. In this 3D co-culture model, both M1 and M2 macrophages showed significantly higher accumulation of Foslip^® compared to the cancer cells. Although this differential accumulation did not drastically affect the overall PDT efficiency, spheroids generated with conditioned media exhibited a significant enhancement in photo-induced cell death, suggesting that the microenvironment could modulate the response to Foslip^®-PDT.

Conclusion: 3D models of HNSCC cells and macrophages provide valuable insights into the complex response of HNSCC cells to PDT using Foslip^® in vitro. This model can be used to screen immunomodulatory nanomedicines targeting TAMs in solid head and neck tumors, either alone or in combination with standard therapies.

Keywords: Foslip; head and neck cancer; macrophage polarization; photodynamic therapy; tumor spheroids.

MeSH terms

Cell Line, Tumor
Culture Media, Conditioned / pharmacology
Head and Neck Neoplasms* / drug therapy
Head and Neck Neoplasms* / pathology
Humans
Macrophages
Squamous Cell Carcinoma of Head and Neck / drug therapy
Squamous Cell Carcinoma of Head and Neck / pathology
Tumor Microenvironment*

Substances

temoporfin
Culture Media, Conditioned