Identifying activators of interferon regulatory factors for neuroprotection

 描述(申请人提供)中风是美国发病率和死亡率的主要原因。然而，只有不到20%的患者有资格接受目前批准的组织型纤溶酶原激活剂或血栓切除术的干预。我们寻求开发新的治疗方法，以减少脑缺血损伤造成的损害和功能障碍的程度，这是一个重要的未得到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，与减少缺血性损伤有关。使用细胞和小鼠中风模型，我们已经证明，在缺血性损伤后给予化合物，已知可以诱导IRF介导的基因转录，显著降低损害的程度。这些结果表明，IRF转录因子的激活 卒中可能是治疗中风患者的一种可行的治疗干预措施。该STTR计划的最终目标是为中风的临床前开发寻找具有最小非靶标免疫活性和对人类和小鼠的特异性的IRF激活剂。我们这一阶段应用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，以便进一步开发。我们提出了以下目标：目的1：验证一种高通量的初步筛选方法，以使用人THP1Dual ISRE/NFκB报告细胞系和384-well格式来鉴定有效和选择性的irf激动子。目的2：验证在含有双重ISRE/NFkB报告基因的小鼠J774细胞中的高通量二次筛选方法，以评估来自目标1的一次HITS的跨物种活性。目标3：通过评估人和小鼠脑源性细胞中一组IRF诱导的下游基因，验证二次HITS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.