IL-23 and IL-17A mRNA-targeted oligonucleotide therapeutics in autoimmune uveitis

促炎症细胞因子IL-23和IL-17A在自身免疫性葡萄膜炎(AU)中起重要作用。 目前AU的治疗方法包括免疫抑制剂，包括全身皮质类固醇， 伴随着许多具有挑战性的毒性。作为一种新的细胞因子基因表达调控 机制，我们发现microRNA(MiRNA)miR4661-3p与mRNA-1协同作用。 稳定蛋白HUR以提高IL-17A的mRNA和蛋白水平。利用这一不同寻常的 作为潜在治疗靶点的miRNA介导的增强机制 Treateutics和耶鲁大学合作开发了一种靶点阻滞剂(TSB) 特异性阻断miR466l-3p与IL-17A 3‘UTR结合的寡核苷酸，导致 在体外和体内，转录产物衰退并降低IL-17A蛋白水平。这包括我们的 TSB在大鼠自身免疫性葡萄膜炎模型中的初步疗效数据。IL-23、α-干扰素的mRNA水平 IL-17A上游的细胞因子也在miR466l过表达的细胞中增强。这一阶段 I SBIR将提供miR4661-细胞因子mRNA相互作用特异性TSB的概念证明 寡核苷酸单独或联合应用将是治疗AU的一种新的有效方法。我们会这样做的 通过：(1)设计、验证和评价miR4661-IL-23 3‘非编码区TSB的特异性； (2)优化基础(大鼠)模型的IL-17A mRNA特异性TSB； IL-17A和IL-23TSB单独及联合应用对大鼠AU的疗效评价 模型，通过临床评分，组织病理学和房水水平的TSB和相关 细胞因子。这一第一阶段SBIR项目可实现的里程碑是&gt；临床减少50% 和组织病理学AU参数，从而提供这种新方法的治疗益处 自身免疫性葡萄膜炎。

The pro-inflammatory cytokines IL-23 and IL-17A play critical roles in autoimmune uveitis (AU). Current AU treatments include immunosuppressants, including systemic corticosteroids, accompanied by many challenging toxicities. As a novel cytokine gene expression control mechanism, we found that the microRNA (miRNA) miR466l-3p cooperates with the mRNA- stabilizing protein HuR to augment IL-17A mRNA and protein levels. To exploit this unusual miRNA-mediated enhancing mechanism as a potential therapeutic target, TargetSite Therapeutics and Yale University have collaboratively generated a target site blocker (TSB) oligonucleotide that specifically blocks miR466l-3p binding to the IL-17A 3’UTR, leading to transcript decay and decreased IL-17A protein levels both in vitro and in vivo. This includes our preliminary TSB efficacy data in the rat autoimmune uveitis model. The mRNA levels of IL23, a cytokine upstream of IL-17A, are also augmented in miR466l-overexpressing cells. This Phase I SBIR will provide proof-of-concept that miR466l – cytokine mRNA interaction-specific TSB oligos, either singly or in combination, will be a novel effective treatment for AU. We will do so by: (1) designing, validating and evaluating specificity of a miR466l – IL-23 3’UTR TSB; (2) optimizing the IL-17A mRNA-specific TSB for the foundational (rat) model; and (3) assessing the efficacy of the IL-17A and IL-23 TSBs, singly and in combination, in the rat AU model, by clinical scoring, histopathology and aqueous humor levels of the TSBs and relevant cytokines. The achievable milestone for this Phase I SBIR project is >50% reduction in clinical and histopathologic AU parameters, thereby offering therapeutic benefit of this novel approach in autoimmune uveitis.