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

基本信息

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
项目类别：
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

项目摘要

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.

概括。我们已经证明，药理学 FAK 抑制和星形胶质细胞 FAK 缺失是对雌性小鼠（而非雄性小鼠）缺血性 MCAO 中风后具有脑保护作用。系统性小分子FAK MCAO 再灌注后 6 小时开始抑制剂治疗是有效的，如运动表现所示损伤部位的功能和组织学分析。我们建议使用 FAK 抑制剂或一种正在进行临床试验的抑制剂癌症作为 SPAN 测试站点临床前评估的候选者，包括在年轻人中的复制小鼠、老年小鼠、大鼠和患有合并症的动物。这一发现是基于十多项研究在我们的实验室发现了调节细胞因子的新整合素-FAK-STAT3 信号通路多年后表达。在雌性小鼠中，血浆玻连蛋白（VTN）渗入脑损伤处，激活整合素，加剧前两天的进行性损伤，这在很大程度上是由急性前体介导的大脑中炎症性 IL-6 的表达。仅女性中风诱发的血浆 VTN 水平和水平与更严重的组织损伤相关。 VTN诱导IL-6机制不受卵巢切除术的影响表明性激素不参与其中。使用 cre-lox 小鼠，我们确定星形胶质细胞 FAK 是主要的雌性小鼠而非雄性小鼠中有害 IL-6 峰值的驱动因素。此外，FAK14 在以下情况下具有脑保护作用： WT 雌性，但不在 VTN-/- 雌性同窝仔或雄性中。因此，我们已经确定了一种多效性机制它可以被有害血液蛋白下游的药物抑制，无论其水平如何。瞄准 1、Hagg实验室将确定两种FAK抑制剂在小鼠中的剂量反应曲线，以确定最低剂量具有最大功效的剂量及其效力。结果测量将是 FAK 磷酸化和管腔内丝状 MCAO 再灌注后 24 小时脑组织中细胞因子的表达 7天时的功能缺陷和脑损伤大小。测试地点将获得两个中最有希望的对我们从供应商处收到的化合物进行质量控制后的抑制剂。对于目标 2a，测试地点将重复我们的发现，即 FAK 抑制剂在年轻成年 C57BL/6J 女性的 MCAO 后具有脑保护作用而不是男性。结果指标包括通过重复 MRI 测量的损伤大小和感觉运动功能根据当前 SPAN 的定义，测试时间超过 30 天。 Aim 2b 将在老年小鼠中进行测试，Aim 2c 将在年轻小鼠中进行测试成年大鼠。根据结果，最常见危险因素合并症的小鼠或大鼠模型人类中风、高血压和糖尿病将在 Aim 2d 中进行测试。为了扩大潜在的临床影响， Aim 2e 将在凝块-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来自试验场啮齿动物的大脑。我们还预计 FAK 抑制剂会降低风险。关键里程碑将选择 FAK 抑制剂及其剂量、复制并权衡其潜在的临床前景鉴于测试点测试的不同模型显示出脑保护作用。如果成功的话，我们将为临床卒中试验提供一种针对特定机制且耐受性良好的新干预措施。