scRNA-seq和蛋白質組學揭示了m2樣巨噬細胞在惡性膠質瘤中關鍵作用

2023年5月，福建醫科大學附屬醫院神經外科;福建醫科大學附屬醫院福建省婦兒醫院婦科;莆田學院附屬醫院神經外科;福州平潭綜合實驗區醫院 (Department of Neurosurgery, Fujian Medical University Union Hospital,Fuzhou, China；Fujian Medical University, Fuzhou, China；Department of Gynaecology, Fujian Provincial Maternity and Children's Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, China；Department of Neurosurgery, Affiliated Hospital of Putian University, Putian,China；Pingtan Comprehensive Experimental Area Hospital, Fuzhou, China) Guiting You老師研究團隊在《CNS Neuroscience & Therapeutics》上發表論文：

“scRNA-seq and proteomics reveal the distinction of M2-like macrophages between primary and recurrent malignant glioma and its critical role in the recurrence"

“scRNA-seq和蛋白質組學揭示了m2樣巨噬細胞在原發性和復發性惡性膠質瘤中的差異及其在復發中的關鍵作用"

Abstract：

Aims: Tumor-associated macrophages (TAMs) in the immune microenvironment play an important role in the increased drug resistance and recurrence of malignant glioma, but the mechanism remains incompletely inventoried. The focus of this study was to investigate the distinctions of M2-like TAMs in the immune microenvironment between primary and recurrent malignant glioma and its influence in the recurrence.

Methods: We employed single-cell RNA sequencing to construct a single-cell atlas for a total of 23,010 individual cells from 6 patients with primary or recurrent malignant glioma and identified 5 cell types, including TAMs and malignant cells. Immunohistochemical techniques and proteomics analysis were performed to investigate the role of intercellular interaction between malignant cells and TAMs in the recurrence of malignant glioma.

Results: Six subgroups of TAMs were annotated and M2-like TAMs were found to increase in recurrent malignant glioma significantly. A pseudotime trajectory and a dynamic gene expression profiling during the recurrence of malignant glioma were reconstructed. Up-regulation of several cancer pathways and intercellular interaction-related genes are associated with the recurrence of malignant glioma. Moreover, the M2-like TAMs can activate the PI3K/Akt/HIF-1α/CA9 pathway in the malignant glioma cells via SPP1-CD44-mediated intercellular interaction. Interestingly, high expression of CA9 can trigger the immunosuppressive response in the malignant glioma, thus promoting the degree of malignancy and drug resistance.

Conclusion: Our study uncovers the distinction of M2-like TAMs between primary and recurrent glioma, which offers unparalleled insights into the immune microenvironment of primary and recurrent malignant glioma.

摘要：

目的:免疫微環境中的腫瘤相關巨噬細胞(Tumor-associated macrophages, tam)在惡性膠質瘤耐藥增加和復發中發揮重要作用，但其機制尚不清楚。本研究的重點是探討原發性和復發性惡性膠質瘤免疫微環境中m2樣tam的差異及其對復發的影響。

方法:采用單細胞RNA測序技術，構建來自6例原發性或復發性惡性膠質瘤患者的23,010個細胞的單細胞圖譜，鑒定出包括tam細胞和惡性細胞在內的5種細胞類型。通過免疫組織化學技術和蛋白質組學分析，研究惡性細胞與tam之間的細胞間相互作用在惡性膠質瘤復發中的作用。

結果:6個tam亞組被注釋，發現復發性惡性膠質瘤中m2樣tam顯著增加。重建惡性膠質瘤復發期間的假時間軌跡和動態基因表達譜。幾種腫瘤通路和細胞間相互作用相關基因的上調與惡性膠質瘤的復發有關。此外，m2樣tam可以通過spp1 - cd44介導的細胞間相互作用激活惡性膠質瘤細胞中的PI3K/Akt/HIF-1α/CA9通路。有趣的是，在惡性膠質瘤中，CA9的高表達可引發免疫抑制反應，從而促進惡性程度和耐藥。

結論:研究人員的研究揭示了原發性和復發性膠質瘤之間m2樣tam的差異，為原發性和復發性惡性膠質瘤的免疫微環境提供了見解。

該論文中，人膠質瘤U87細胞及其慢病毒轉染細胞的體外培養是使用Ausbian特級胎牛血清完成的。