Laboratory of Experimental Neurology and Neuroimmunology, Second Department of Neurology, AHEPA University Hospital, Thessaloniki, Greece.
Department of Neuroscience, Central Clinical School, Monash University, Prahran, Victoria, Australia.
Brain Pathol. 2022 Jul;32(4):e13040. doi: 10.1111/bpa.13040. Epub 2021 Nov 29.
Experimental autoimmune encephalomyelitis (EAE) is a basic and reliable model used to study clinical and pathological hallmarks of multiple sclerosis (MS) in rodents. Several studies suggest neural precursor cells (NPCs) as a significant research tool while reporting that transplanted NPCs are a promising therapeutic approach to treating neurological disorders, such as MS. The main objective was to approach a preclinical, in vivo scenario of oligodendrogenesis with NPCs, targeting the main chronic demyelinated lumbosacral milieu of EAE, via the least invasive delivery method which is lumbar puncture. We utilized MOG peptide to induce EAE in C57BL/6 mice and prior to the acute relapse, we intervened with either the traceable GFP cellular therapy or saline solution in the intrathecal space of their lumbar spine. A BrdU injection, which enabled us to monitor endogenous proliferation, marked the endpoint 50 days post-induction (50 dpi). Neuropathology with high-throughput, triple immunofluorescent, and transmission electron microscopy (TEM) data were extracted and analyzed. The experimental treatment attenuated the chronic phase of EAE (50 dpi; score <1) following an acute, clinical relapse. Myelination and axonal integrity were rescued in the NPC-treated animals along with suppressed immune populations. The differentiation profile of the exogenous NPCs and endogenous BrdU cells was location-dependent where GFP -rich areas drove undifferentiated phenotypes toward the oligodendrocyte lineage. In situ oligodendrocyte enrichment was demonstrated through increased (p < 0.001) gap junction channels of Cx32 and Cx47, reliable markers for proliferative oligodendroglia syncytium. TEM morphometric analysis ultimately manifested an increased g-ratio in lumbosacral fibers of the recovered animals (p < 0.001). Herein, we suggest that a single, lumbar intrathecal administration of NPCs capacitated a viable cellular load and resulted in clinical and pathological amelioration, stimulating resident OPCs to overcome the remyelination failure in EAE demyelinating locale.
实验性自身免疫性脑脊髓炎 (EAE) 是一种基础且可靠的模型，用于在啮齿动物中研究多发性硬化症 (MS) 的临床和病理学特征。有几项研究表明神经前体细胞 (NPC) 是一种重要的研究工具，同时报告说移植 NPC 是治疗神经紊乱（如 MS）的一种很有前途的治疗方法。主要目标是通过最微创的递送方法（即腰椎穿刺）接近 NPC 的少突胶质前体细胞的临床前、体内情景，针对 EAE 的主要慢性脱髓鞘腰骶环境。我们利用 MOG 肽在 C57BL/6 小鼠中诱导 EAE，并在急性复发之前，在其腰椎的鞘内空间中用可追踪的 GFP 细胞治疗或生理盐水进行干预。BrdU 注射使我们能够监测内源性增殖，并在诱导后 50 天（50 dpi）标记终点。提取和分析了具有高通量、三重免疫荧光和透射电子显微镜 (TEM) 数据的神经病理学。实验治疗减轻了急性临床复发后的 EAE 慢性期（50 dpi；评分 <1）。在 NPC 治疗的动物中，髓鞘形成和轴突完整性得到了恢复，免疫群体受到抑制。外源性 NPC 和内源性 BrdU 细胞的分化谱与位置有关，其中富含 GFP 的区域将未分化表型推向少突胶质细胞谱系。通过增加（p <0.001）缝隙连接通道 Cx32 和 Cx47 证明了原位少突胶质细胞富集，这是增殖性少突胶质细胞合胞体的可靠标志物。TEM 形态计量分析最终表明，恢复动物腰骶纤维中的 g-比值增加（p <0.001）。在此，我们认为 NPC 的单次腰椎鞘内给药可提供可行的细胞负荷，并导致临床和病理改善，刺激驻留的 OPC 克服 EAE 脱髓鞘部位的再髓鞘失败。
