Translational assessment of an FAK inhibitor for acute cerebroprotection

FAK 抑制剂急性脑保护作用的转化评估

• 批准号: 10673417
• 负责人: THEO HAGG
• 金额: $ 30.18万
• 依托单位: EAST TENNESSEE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10673417
• 关键词: Acute; Adjuvant; Affect; Agreement; Animals; Astrocytes; Behavioral; Blood; Blood Proteins; Brain; Brain Injuries; Clinical; Coagulation Process; Deposition; Diabetes Mellitus; Disease; Dose; Enzyme-Linked Immunosorbent Assay; Exclusion; Female; Filament; Genetic; Gonadal Steroid Hormones; Grant; Histologic; Histology; Human; Hypertension; Image; Inflammatory; Injury; Integrins; Interleukin-6; Intervention; Ischemia; Ischemic Stroke; Lead; Light; Magnetic Resonance Imaging; Malignant Neoplasms; Measures; Mediating; Methods; Modeling; Motor; Mus; National Institute of Neurological Disorders and Stroke; Neurologic Deficit; Outcome; Outcome Measure; Ovariectomy; PTK2 gene; Paper; Pharmaceutical Preparations; Phosphorylation; Plasma; Quality Control; Rattus; Reperfusion Therapy; Risk; Risk Factors; Rodent; Rodent Model; Safety; Sensorimotor functions; Sex Differences; Signal Transduction; Site; Specificity; Sprague-Dawley Rats; Stat3 Signaling Pathway; Stroke; Testing; Time; Tissues; Toxic effect; Vitronectin; Walking; Woman; Work; aged; brain tissue; cancer clinical trial; cerebroprotection; clinically relevant; comorbidity; cytokine; efficacy evaluation; improved outcome; inhibitor; mRNA Expression; male; pharmacologic; phase II trial; pre-clinical assessment; response; sex; small molecule; standard of care; stroke clinical trials; stroke outcome; stroke victims; support network; tissue injury; young adult

Summary. We have shown that a pharmacological FAK inhibition and astrocyte FAK deletion are cerebroprotective after ischemic MCAO stroke in female, but not male, mice. A systemic small molecule FAK inhibitor treatment was efficacious when started 6 h after reperfusion following an MCAO, as shown by motor function and histological analyses of the injury site. We propose the FAK inhibitor or one in clinical trials for cancer as a candidate for preclinical assessment by the SPAN test sites, including replication in young adult mice, in aged mice, in rats and animals with comorbidities. This finding is based on work over more than ten years in our lab after we discovered a new integrin-FAK-STAT3 signaling pathway that regulates cytokine expression. In female mice, plasma vitronectin (VTN) that leaks into the brain injury activates integrins to exacerbate the progressive injury over the first two days which is mediated in large part by acute pro- inflammatory IL-6 expression in the brain. Stroke induced plasma VTN levels in females only and the levels correlate with worse tissue injury. The VTN-induced IL-6 mechanism was not affected by ovariectomy suggesting that sex hormones are not involved. Using cre-lox mice, we identified astroglial FAK as the major driver of the detrimental IL-6 peak in female, but not male, mice. Moreover, FAK14 was cerebroprotective in WT females but not in VTN-/- female littermates or in males. Thus, we have identified a pleiotropic mechanism that can be inhibited by a drug downstream of a detrimental blood protein and irrespective of its levels. In Aim 1, the Hagg lab will determine a dose-response curve in mice for the two FAK inhibitors to define the lowest dose that has maximal efficacy, and their potency. Outcome measures will be FAK phosphorylation and cytokine expression in brain tissue at 24 h after intraluminal filament MCAO with reperfusion and for reducing functional deficits and brain injury size at 7 d. The test sites would receive the most promising of the two inhibitors after quality control for the compounds we receive from suppliers. For Aim 2a, the test sites would replicate our finding that the FAK inhibitor is cerebroprotective after MCAO in young adult C57BL/6J females and not males. Outcome measures are injury size, as measured by repeated MRI, and sensorimotor function tests over 30 days, as defined by the current SPAN. Aim 2b would test it in aged mice and Aim 2c in young adult rats. Depending on the outcome, mouse or rat models of the most common risk factor comorbidities of human stroke, hypertension and diabetes, will be tested in Aim 2d. To broaden the potential clinical impact, Aim 2e will test the FAK inhibitor in a clot-tPA reperfusion model. In Aim 3, our lab will determine whether high VTN levels caused by comorbidities are a risk factor for worse stroke outcomes by analyzing plasma and brains from test site rodents. We also expect the risk to be reduced by the FAK inhibitor. Key milestones will be the selection of the FAK inhibitor and its dosing, replication and weighing its potential clinical promise in light of showing cerebroprotective effects in different models tested by the testing sites. If successful, we will contribute a new intervention which targets a specific mechanism and is well-tolerated, for clinical stroke trials.

总结。我们已经证明，药物对FAK的抑制和星形胶质细胞FAK的缺失 雌性小鼠脑缺血后的脑保护作用，但雄性小鼠无此作用。一种系统性小分子FAK 运动试验显示，在大脑中动脉闭塞后再灌注6小时后开始使用抑制剂是有效的。 损伤部位的功能及组织学分析。我们建议使用FAK抑制剂或在临床试验中使用 SPAN测试点将癌症作为临床前评估的候选对象，包括年轻人的复制 小鼠，老年小鼠，大鼠和患有共病的动物。这一发现是基于十多年来的工作 在我们的实验室中，我们发现了一条新的整合素-FAK-STAT3信号通路来调节细胞因子 表情。在雌性小鼠中，泄漏到脑损伤中的血浆玻璃连结蛋白(VTN)激活整合素以 在头两天加重进行性损伤，这在很大程度上是由急性亲和力调节的 炎症性IL-6在脑内的表达。仅女性卒中诱发的血浆VTN水平和 与更严重的组织损伤有关。卵巢切除不影响VTN诱导的IL-6机制 这表明性激素与此无关。在cre-lox小鼠身上，我们发现星形胶质细胞FAK是主要的 雌性小鼠体内有害的IL-6峰值的驱动因素，而雄性小鼠则不然。此外，FAK14还具有脑保护作用。 WT雌性，但不在VTN-/-雌性窝产仔或雄性。因此，我们确定了一种多效性机制。 这可以被有害血液蛋白下游的药物抑制，而无论其水平如何。在AIM 1，Hagg实验室将在小鼠中确定两种FAK抑制剂的剂量-反应曲线，以确定最低 具有最大疗效的剂量及其效力。结果指标将是FAK磷酸化和 MCAO再灌流后24小时脑组织细胞因子的表达及意义 7天时的功能缺陷和脑损伤大小。测试地点将收到两个地点中最有希望的 对我们从供应商那里收到的化合物进行质量控制后使用抑制剂。对于AIM 2a，测试地点将 重复我们的发现，FAK抑制剂对年轻成年C57BL/6J女性大脑中动脉闭塞后具有脑保护作用 而不是男性。结果：通过重复的核磁共振测量损伤的大小和感觉运动功能。 测试持续30天，由当前跨度定义。目标2b将在老年小鼠身上进行测试，目标2c将在年轻小鼠身上进行测试 成年大鼠。根据结果，最常见的危险因素并存的小鼠或大鼠模型 人类中风、高血压和糖尿病将在Aim 2d进行测试。为了扩大潜在的临床影响， Aim 2E将在血栓-tPA再灌注模型中测试FAK抑制剂。在目标3中，我们的实验室将确定是否高 通过分析血浆和脑脊液中的VTN水平，发现由合并症引起的VTN水平是导致更差的中风预后的危险因素 来自试验场啮齿动物的大脑。我们还预计FAK抑制剂可以降低风险。关键里程碑将 FAK抑制剂的选择及其剂量、复制和权衡其潜在的临床前景 在测试现场测试的不同型号中显示出脑保护效果。如果成功，我们将 为临床中风试验贡献一种针对特定机制且耐受性良好的新干预措施。