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。我们已经表明，药理学FAK抑制和星形胶质细胞FAK缺失是雌性的缺血性MCA中风后，但不是雄性小鼠的脑脑保护。全身小分子fak MCAO后6小时开始，抑制剂治疗是有效的，如马达所示损伤部位的功能和组织学分析。我们提出了FAK抑制剂或在临床试验中的一种癌症作为跨越测试地点的临床前评估的候选者，包括年轻人的复制在老年小鼠中，有合并症的大鼠和动物的小鼠。这一发现是基于十多次的工作在我们发现了一种调节细胞因子的新的整合素fak-stat3信号传导途径之后，我们在实验室里的几年表达。在雌性小鼠中，泄漏到脑损伤的血浆玻璃体（VTN）激活整联蛋白至加剧了最初两天的进行性损伤大脑中的炎症IL-6表达。中风诱导女性的血浆VTN水平和水平与较差的组织损伤相关。 VTN诱导的IL-6机制不受卵巢切除术的影响表明性激素不涉及。使用Cre-lox小鼠，我们确定了星形胶质体FAK为主要女性而不是雄性小鼠的有害IL-6峰的驱动器。此外，FAK14在 wt女性，但不在VTN - / - 女性同窝室或男性中。因此，我们已经确定了一种多效机制这可以被有害血蛋白下游的药物抑制，而其水平不管其水平如何。目标 1，HAGG实验室将确定两个FAK抑制剂的小鼠中的剂量反应曲线以定义最低具有最大功效的剂量及其效力。结果指标将是FAK磷酸化和腔内丝MCAO后24小时内的细胞因子表达，并减少 7 d时功能缺陷和脑损伤大小。测试站点将获得两者中最有希望的我们从供应商那里收到的化合物的质量控制后的抑制剂。对于AIM 2A，测试站点将复制我们的发现，即FAK抑制剂在MCAO之后在年轻的成人C57BL/6J女性中受到脑外保护而不是男性。结果指标是损伤的大小，通过重复MRI和感觉运动功能测量测试超过30天，如当前跨度所定义。 AIM 2B将在老年小鼠中对其进行测试，而AIM 2C在Young中进行测试成年大鼠。取决于最常见危险因素合并症的结果，鼠标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AIM 3中，我们的实验室将确定是否高合并症引起的VTN水平是通过分析血浆和血浆和测试现场啮齿动物的大脑。我们还期望FAK抑制剂降低风险。关键的里程碑将选择FAK抑制剂及其给药，复制并权衡其潜在的临床承诺在测试位点测试的不同模型中显示脑保护作用的光。如果成功，我们将为临床中风试验做出针对特定机制并具有良好耐受性的新干预措施。