Identifying activators of interferon regulatory factors for neuroprotection

 描述（由申请人提供）中风是美国发病率和死亡率的主要原因。然而，不到20%的患者有资格接受目前批准的组织纤溶酶原激活剂干预或血栓切除术。我们寻求开发新的治疗方法，以减少大脑缺血性损伤导致的损伤和功能障碍的程度，这是一个显著未满足的医疗需求领域。我们发现干扰素调节因子（IRF）介导的基因转录可能代表了一种内源性神经保护机制，与缺血性损伤的减少有关。使用中风的细胞和小鼠模型，我们已经证明在缺血性损伤后施用已知诱导IRF介导的基因转录的化合物显著降低损伤的程度。这些结果表明，IRF转录因子的激活， 中风可能是治疗中风患者的可行的治疗干预。该STTR项目的最终目标是鉴定对小鼠和人类具有最小脱靶免疫活性和特异性的IRF激活剂，用于卒中的临床前开发。我们的第一阶段申请的具体目标是开发和验证一个高通量筛选平台，以确定临床上可行的IRF激活化合物，以进一步开发。我们提出以下目标：目标1：使用人THP 1双重ISRE/NFκB报告细胞系和384孔板，进行高通量初步筛选试验，以鉴定强效和选择性IRF激活剂。目标二：在含有双重ISRE/NFkB报告基因的小鼠J774细胞中进行高通量二次筛选测定，以评估来自Aim 1的初次命中的跨物种活性。目标3：通过评估人和小鼠脑源性细胞中的一组下游IRF诱导基因，验证二次命中的靶点和物种选择性。

 DESCRIPTION (provided by applicant) Stroke is a leading cause of morbidity and mortality in the United States. However less than 20% of patients are eligible for the current approved interventions of tissue plasminogen activator or thrombectomy. We seek to develop new therapeutics to reduce the extent of damage and functional impairment resulting from ischemic injury to the brain, an area of significant unmet medical need. We have found that interferon regulatory factor (IRF) mediated gene transcription may represent an endogenous mechanism of neuroprotection that is associated with a reduction in ischemic injury. Using both cell and mouse models of stroke we have demonstrated that administration of compounds following the ischemic insult, that are known to induce IRF mediated gene transcription significantly reduces the extent of damage. These results indicate that activation of IRF transcription factors following stroke may be a viable therapeutic intervention for the treatment of stroke patients. The ultimate goal of this STTR program is to identify IRF activators with minimal off-target immune activity and specificity for both mouse and human for preclinical development for stroke. Our specific goal for this Phase I application is to develop and validate a high-throughput screening platform to identify clinically viable IRF activating compounds for further development. We propose the following aims: Aim 1: Validate a high-throughput primary screening assay to identify potent and selective IRF activators using a human THP1 dual ISRE/NFκB reporter cell line and 384-well format. Aim 2: Validate a high-throughput secondary screening assay in mouse J774 cells containing dual ISRE/NFkB reporters to evaluate cross-species activity of primary hits from Aim 1. Aim 3: Verify the target and species selectivity of secondary hits by evaluating a panel of downstream IRF-inducible genes in human and mouse brain-derived cells.