Research Department, Institut de Cancérologie de Lorraine, Vandoeuvre-lès-Nancy, France.
Centre de Recherche en Automatique de Nancy, Centre National de la Recherche Scientifique, UMR 7039, Université de Lorraine, Vandoeuvre-lès-Nancy, France.
Int J Nanomedicine. 2023 Nov 9;18:6545-6562. doi: 10.2147/IJN.S427350. eCollection 2023.
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.
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.
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.
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.
肿瘤微环境（TME）由各种基质成分组成，包括肿瘤相关巨噬细胞（TAMs）等免疫细胞，它们在癌症的发生和发展中起着关键作用。TAMs 可表现为抑瘤性 M1 或促瘤性 M2 表型。首先，我们旨在开发一种由头颈部鳞状细胞癌（HNSCC）细胞和不同亚型的巨噬细胞组成的 3D 人异质模型，以复制免疫细胞与癌细胞之间的相互作用。我们进一步研究了脂质体载体制剂 temoporfin（Foslip）在富含巨噬细胞的 3D 模型中的行为。
单核细胞分化为 M1 和 M2 巨噬细胞，这两种巨噬细胞代表两种截然不同的亚型。在进行组织学和分子特征分析后，将这些巨噬细胞用于建立富含极化巨噬细胞或条件培养基的 HNSCC 3D 球体模型。使用流式细胞术和荧光显微镜评估 Foslip 的积累和分布。使用流式细胞术评估 Foslip 介导的光动力疗法（PDT）的细胞毒性作用。
我们开发了具有均匀分布的巨噬细胞混合表型的异质球体。在这种 3D 共培养模型中，M1 和 M2 巨噬细胞对 Foslip 的积累明显高于癌细胞。尽管这种差异积累并没有极大地影响整体 PDT 效率，但用条件培养基生成的球体显示出光诱导细胞死亡的显著增强，这表明微环境可以调节对 Foslip-PDT 的反应。
HNSCC 细胞和巨噬细胞的 3D 模型提供了有价值的见解，可了解体外使用 Foslip 进行 PDT 时 HNSCC 细胞的复杂反应。该模型可用于筛选针对实体头颈部肿瘤中 TAMs 的免疫调节纳米药物，无论是单独使用还是与标准疗法联合使用。
