Dao Nyesiga Gillian, Haslund-Vinding Jeppe Lohfert, Budde Josephine, Lange Josefine Føns, Blum Nadja, Dukstaite Kotryna, Ohlsson Lars, Mathiesen Tiit, Woetmann Anders, Vilhardt Frederik
Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, 205 06 Malmo, Sweden.
Department of Neurosurgery, Copenhagen University Hospital-Rigshospitalet, 2100 Copenhagen, Denmark.
Cancers (Basel). 2024 Nov 25;16(23):3942. doi: 10.3390/cancers16233942.
Current challenges in meningioma treatment, including post-surgical complications and cognitive impairments, highlight the need for new treatment alternatives. Immunological interventions have shown promise. However, there is a knowledge gap in characterizing infiltrating immune cells in meningioma and their interplay. Further studies on immune cells in single-cell suspensions from digested meningioma tissues could identify targetable mechanisms for non-surgical treatment options with fewer side effects. This study aimed to optimize a protocol for faster digestion of meningioma tissues into viable single-cell suspensions and to identify infiltrating immune cell populations.
We modified a commercial kit intended for whole skin dissociation to digest resected meningioma tissues into viable single-cell suspensions. Tumor-infiltrating immune cell populations were characterized using flow cytometry.
Flow cytometry analyses revealed that the digested tissue was composed of viable immune cells, including predominantly CD14 macrophages and CD3 T cells, with minor populations of CD56 NK cells and CD19 B cells. In both of the two patient samples tested, half of the tumor-associated macrophages were TIM-3, with a small proportion co-expressing CD83. Women were more likely to have a lower proportion of immune cells, B cells, and NK cells. Female patients with a high proportion of immune cells had a higher proportion of macrophages.
We successfully optimized a protocol for generating single-cell suspensions with viable immune cells from meningioma tissues, revealing infiltrating antigen-presenting cells with an immunosuppressive phenotype, and lymphocytes. This short protocol allows advanced analyses of tumor-infiltrating cells using techniques such as single-cell RNA sequencing and flow cytometry, which require live, dissociated cells.
脑膜瘤治疗目前面临的挑战,包括术后并发症和认知障碍,凸显了对新治疗方案的需求。免疫干预已显示出前景。然而,在表征脑膜瘤中浸润的免疫细胞及其相互作用方面存在知识空白。对消化后的脑膜瘤组织单细胞悬液中的免疫细胞进行进一步研究,可能会确定副作用较少的非手术治疗选择的可靶向机制。本研究旨在优化一种方案,以便更快地将脑膜瘤组织消化成有活力的单细胞悬液,并确定浸润的免疫细胞群体。
我们修改了一种用于全皮肤解离的商业试剂盒,以将切除的脑膜瘤组织消化成有活力的单细胞悬液。使用流式细胞术对肿瘤浸润免疫细胞群体进行表征。
流式细胞术分析显示,消化后的组织由有活力的免疫细胞组成,主要包括CD14巨噬细胞和CD3 T细胞,还有少量的CD56 NK细胞和CD19 B细胞。在测试的两个患者样本中,一半的肿瘤相关巨噬细胞为TIM-3,一小部分共表达CD83。女性的免疫细胞、B细胞和NK细胞比例更可能较低。免疫细胞比例高的女性患者巨噬细胞比例更高。
我们成功优化了一种从脑膜瘤组织生成含有有活力免疫细胞的单细胞悬液的方案,揭示了具有免疫抑制表型的浸润抗原呈递细胞和淋巴细胞。这个简短的方案允许使用单细胞RNA测序和流式细胞术等技术对肿瘤浸润细胞进行深入分析这些技术需要活的、解离的细胞。