Andersen Brian M, Faust Akl Camilo, Wheeler Michael A, Li Zhaorong, Diebold Martin, Kilian Michael, Rone Joseph M, Misra Aditya, Kenison Jessica E, Lee Joon-Hyuk, Lee Hong-Gyun, Polonio Carolina M, Merrell David, Weiss Jakob H, Godinez Lillie, Piester Gavin, Illouz Tomer, Ye Jessica J, Ghia Arianna, Martinez Jazmin, Chung Elizabeth N, Srun Lena, Farrenkopf Daniel, Flausino Lucas E, Schüle Anton M, Sanmarco Liliana M, Giovannoni Federico, Fehrenbacher Luca, Charabati Marc, Gutiérrez-Vázquez Cristina, Cusick Margaret M, Prabhakar Prem S, Bossi Connor C, Lapinskas Emily, Nowarski Roni, Getz Gad, Ligon Keith L, Prinz Marco, Chiocca E Antonio, Reardon David A, Quintana Francisco J
Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
Department of Neurology, Veterans Affairs Medical Center, Harvard Medical School, Jamaica Plain, MA, USA.
Nature. 2025 Jun 25. doi: 10.1038/s41586-025-09191-9.
Glioblastoma (GBM) is the most lethal primary brain malignancy. Immunosuppression in the GBM tumour microenvironment (TME) is an important barrier to immune-targeted therapies, but our understanding of the mechanisms of immune regulation in the GBM TME is limited. Here we describe a viral barcode interaction-tracing approach to analyse TME cell-cell communication in GBM clinical samples and preclinical models at single-cell resolution. We combine it with single-cell and bulk RNA-sequencing analyses, human organotypic GBM cultures, in vivo cell-specific CRISPR-Cas9-driven genetic perturbations as well as human and mouse experimental systems to identify an annexin A1-formyl peptide receptor 1 (ANXA1-FPR1) bidirectional astrocyte-GBM communication pathway that limits tumour-specific immunity. FPR1 inhibits immunogenic necroptosis in tumour cells, and ANXA1 suppresses NF-κB and inflammasome activation in astrocytes. ANXA1 expression in astrocytes and FPR1 expression in cancer cells are associated with poor outcomes in individuals with GBM. The inactivation of astrocyte-glioma ANXA1-FPR1 signalling enhanced dendritic cell, T cell and macrophage responses, increasing infiltration by tumour-specific CD8 T cells and limiting T cell exhaustion. In summary, we have developed a method to analyse TME cell-cell interactions at single-cell resolution in clinical samples and preclinical models, and used it to identify bidirectional astrocyte-GBM communication through ANXA1-FPR1 as a driver of immune evasion and tumour progression.
胶质母细胞瘤（GBM）是最致命的原发性脑恶性肿瘤。GBM肿瘤微环境（TME）中的免疫抑制是免疫靶向治疗的重要障碍，但我们对GBM TME中免疫调节机制的了解有限。在此，我们描述了一种病毒条形码相互作用追踪方法，以单细胞分辨率分析GBM临床样本和临床前模型中的TME细胞间通讯。我们将其与单细胞和批量RNA测序分析、人器官型GBM培养、体内细胞特异性CRISPR-Cas9驱动的基因扰动以及人和小鼠实验系统相结合，以确定一种膜联蛋白A1-甲酰肽受体1（ANXA1-FPR1）双向星形胶质细胞-GBM通讯途径，该途径限制肿瘤特异性免疫。FPR1抑制肿瘤细胞中的免疫原性坏死性凋亡，而ANXA1抑制星形胶质细胞中的NF-κB和炎性小体激活。星形胶质细胞中ANXA1的表达和癌细胞中FPR1的表达与GBM患者的不良预后相关。星形胶质细胞-胶质瘤ANXA1-FPR1信号的失活增强了树突状细胞、T细胞和巨噬细胞的反应，增加了肿瘤特异性CD8 T细胞的浸润并限制了T细胞耗竭。总之，我们开发了一种方法，以单细胞分辨率分析临床样本和临床前模型中的TME细胞间相互作用，并利用它来确定通过ANXA1-FPR1的双向星形胶质细胞-GBM通讯是免疫逃逸和肿瘤进展的驱动因素。
