Ji Kyungmin, Schwenkel George J, Mattingly Raymond R, Sundararaghavan Harini G, Zhang Zheng Gang, Chopp Michael
Department of Neurology, Henry Ford Health, Detroit, MI 48202, USA.
Department of Pharmacology and Toxicology, Brody Medical School at East Carolina University, Greenville, NC 27834, USA.
Cancers (Basel). 2024 Jul 9;16(14):2498. doi: 10.3390/cancers16142498.
Plexiform neurofibromas (PNs) occur in about a half of neurofibromatosis type 1 (NF1) patients and have garnered significant research attention due to their capacity for growth and potential for malignant transformation. NF1 plexiform neurofibroma (pNF1) is a complex tumor composed of Schwann cell-derived tumor cells () and the tumor microenvironment (TME). Although it has been widely demonstrated that the TME is involved in the formation of neurofibromas, little is known about the effects of the TME on the subsequent progression of human pNF1. Elucidating the molecular interactions between tumor cells and the TME may provide new therapeutic targets to reduce the progression of pNF1. In the present study, we focused on the contributions of fibroblasts, the most abundant cell types in the TME, to the growth of pNF1. To simulate the TME, we used a three-dimensional (3D) coculture model of immortalized pNF1 tumor cells () and primary fibroblasts () derived from pNF1 patients. We performed live-cell imaging of 3D/4D (3D in real-time) cultures through confocal microscopy followed by 3D quantitative analyses using advanced imaging software. The growth of pNF1 spheroids in 3D cocultures with fibroblasts was significantly greater than that of pNF1 spheroids in 3D monocultures. An increase in the growth of pNF1 spheroids also occurred when they were cultured with conditioned media (CM) from fibroblasts. Moreover, fibroblast-derived CM increased the invasive outgrowth and further local invasion of pNF1 spheroids. Interestingly, when small extracellular vesicles (sEVs) were depleted from the fibroblast-derived CM, the stimulation of the growth of pNF1 spheroids was lost. Our results suggest that fibroblast-derived sEVs are a therapeutic target for reducing the growth of pNF1.
丛状神经纤维瘤（PNs）发生于约一半的1型神经纤维瘤病（NF1）患者中，因其生长能力和恶性转化潜力而受到了大量研究关注。NF1丛状神经纤维瘤（pNF1）是一种由施万细胞来源的肿瘤细胞（）和肿瘤微环境（TME）组成的复杂肿瘤。尽管已有广泛研究表明TME参与神经纤维瘤的形成，但对于TME对人pNF1后续进展的影响却知之甚少。阐明肿瘤细胞与TME之间的分子相互作用可能会提供新的治疗靶点，以减少pNF1的进展。在本研究中，我们聚焦于TME中最丰富的细胞类型——成纤维细胞对pNF1生长的作用。为了模拟TME，我们使用了永生化pNF1肿瘤细胞（）和源自pNF1患者的原代成纤维细胞（）的三维（3D）共培养模型。我们通过共聚焦显微镜对3D/4D（实时3D）培养物进行活细胞成像，随后使用先进的成像软件进行3D定量分析。与成纤维细胞共培养的3D条件下pNF1球体的生长显著大于3D单培养条件下pNF1球体的生长。当pNF1球体与成纤维细胞的条件培养基（CM）一起培养时，其生长也会增加。此外，成纤维细胞来源的CM增加了pNF1球体的侵袭性生长和进一步的局部侵袭。有趣的是，当成纤维细胞来源的CM中的小细胞外囊泡（sEVs）被耗尽时，对pNF1球体生长的刺激作用就消失了。我们的结果表明，成纤维细胞来源的sEVs是减少pNF1生长的一个治疗靶点。
