Department of Health Sciences, University of Genoa, via A. Pastore, 1, 16132, Genoa, Italy.
Hospital Policlinico San Martino-IRCCS-Genova, Genoa, Italy.
Drug Deliv Transl Res. 2018 Oct;8(5):1345-1354. doi: 10.1007/s13346-018-0535-3.
Oligonucleotide overloading results in type I interferonopathies such as the Aicardi-Goutiéres Syndrome, a progressive encephalopathy determined by an immune response against endogenous DNA/RNA molecules. No therapy targeting pathogenic mechanisms is available for affected patients. Accordingly, we set up an in vitro/in vivo experimental model aimed at reproducing the pathogenic mechanisms of type I interferonopathies, in order to develop an effective pharmacological modulation and toxicological alterations caused by intracranial delivery of encapsulated CpG. The in vitro model used Aicardi-Goutiéres Syndrome immortalized lymphocytes activated by interferon I and co-cultured with human astrocytes; lymphocyte neurotoxicity was attenuated by the calcineurin-inhibitor Tacrolimus and by the anti-interferon monoclonal antibody Sifalimumab. The in vivo model was set up in mice by subcutaneous injection of encapsulated CpG oligonucleotides; the immune-stimulating activity was demonstrated by cytometric analysis in the spleen. To mime pathogenesis of type I interferonopathies in the central nervous system, CpG oligonucleotides were administered intracranially in mice. In the brain, CpG overload induced a rapid activation of macrophage-like microglial cells and focal accumulation mononuclear cells. The subcutaneous administration of Tacrolimus and, more potently, Sifalimumab attenuated CpG-induced brain alterations. These findings shed light on molecular mechanisms triggered by oligonucleotides to induce brain damage. Monoclonal antibodies inhibiting interferon seem a promising therapeutic strategy to protect brain in type I interferonopathies.
寡核苷酸过载会导致 I 型干扰素病,如 Aicardi-Goutières 综合征,这是一种由内源性 DNA/RNA 分子引起的免疫反应所决定的进行性脑病。目前尚无针对致病机制的治疗方法。因此,我们建立了一个体外/体内实验模型,旨在复制 I 型干扰素病的致病机制,以便开发针对包裹 CpG 颅内给药引起的有效药物调节和毒性改变。体外模型使用干扰素 I 激活的 Aicardi-Goutières 综合征永生化淋巴细胞,并与人星形胶质细胞共培养;钙调神经磷酸酶抑制剂他克莫司和抗干扰素单克隆抗体 Sifalimumab 可减轻淋巴细胞神经毒性。体内模型通过皮下注射包裹的 CpG 寡核苷酸在小鼠中建立;在脾脏中的细胞计量分析证明了免疫刺激活性。为了模拟 I 型干扰素病在中枢神经系统中的发病机制,将 CpG 寡核苷酸颅内给药于小鼠。在大脑中,CpG 过载会迅速激活巨噬细胞样小胶质细胞,并导致单核细胞局灶性聚集。他克莫司和更有效的 Sifalimumab 皮下给药可减轻 CpG 诱导的脑改变。这些发现揭示了寡核苷酸引发脑损伤的分子机制。抑制干扰素的单克隆抗体似乎是保护 I 型干扰素病中大脑的一种有前途的治疗策略。