MMPs and NETs are detrimental in CNS-tuberculosis with MMP inhibitor doxycycline improving murine CNS-tuberculosis survival (human)

Despite effective anti-tuberculous treatment (ATT), up to 40% of patients with central nervous system tuberculosis (CNS-TB) suffer permanent neurological deficits and death. The immunopathological mechanism of CNS-TB remains undefined, hindering the development of new CNS-TB therapies. We here report the spatial transcriptomics profiling of human CNS-TB and demyelination CNS tissues using laser capture microdissection RNA sequencing (LCM-RNA Seq). Human CNS-TB and demyelination samples exhibited distinct gene expression profiles based on cellular phenotypes. Genes involved in biological processes relates to cytokine responses, cell-substrate adhesion, extracellular matrix organization, inflammatory and immune responses are highly expressed in both inflammatory and granuloma tissues in contrast to demyelination and normal tissues. Down-regulated genes are associated with CNS-related processes such as synapse and cell junction organisation, myelination, neuron development and differentiation. Overall design: Clinical formalin-fixed paraffin-embedded (FFPE) samples were retrieved from human CNS-TB (n=6) and demyelination control (n=4). All tissue sections with 7 µm thickness each were stained with hematoxylin prior to LCM-RNA Seq. Target regions including granuloma, inflammatory region and adjacent normal regions were selected and microdissected for library construction. The libraries were sequenced and analysed.

尽管接受有效的抗结核治疗（anti-tuberculous treatment, ATT），仍有高达40%的中枢神经系统结核（central nervous system tuberculosis, CNS-TB）患者会遗留永久性神经功能缺损甚至死亡。目前中枢神经系统结核的免疫病理机制尚未明确，这阻碍了新型中枢神经系统结核治疗手段的研发。本研究通过激光捕获显微切割RNA测序（laser capture microdissection RNA sequencing, LCM-RNA Seq），对人类中枢神经系统结核及脱髓鞘中枢神经系统组织开展了空间转录组谱分析。人类中枢神经系统结核与脱髓鞘样本基于细胞表型呈现出显著不同的基因表达谱。相较于脱髓鞘组织与正常组织，炎症组织与肉芽肿组织中高表达的基因主要参与细胞因子应答、细胞-基质黏附、细胞外基质组织、炎症与免疫应答等生物学过程。而下调表达的基因则与中枢神经系统相关的生物学过程密切相关，包括突触与细胞连接构建、髓鞘形成、神经元发育与分化等。总体实验设计：本研究收集了人类中枢神经系统结核患者（n=6）与脱髓鞘对照受试者（n=4）的福尔马林固定石蜡包埋（formalin-fixed paraffin-embedded, FFPE）组织样本。所有厚度为7微米的组织切片均在进行激光捕获显微切割RNA测序前用苏木精染色。研究人员选取了包括肉芽肿区域、炎症区域及邻近正常区域在内的目标区域，进行显微切割以构建测序文库。随后对构建完成的文库进行测序与数据分析。