Targeting carbonic anhydrases in Alzheimer's disease

靶向碳酸酐酶治疗阿尔茨海默病

基本信息

批准号：
10374878
负责人：
Silvia Fossati
金额：
$ 45.23万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-15 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10374878
关键词：
3xTg-AD mouse AD transgenic mice Acetazolamide Age Age-Months Aging Altitude Altitude Sickness Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease patient Alzheimer&apos s disease therapy Amyloid Amyloid beta-Protein Amyloidosis Animal Model Behavioral Bicarbonates Biochemical Blood Vessels Brain CASP3 gene Carbon Dioxide Carbonic Anhydrase II Carbonic Anhydrase Inhibitors Caspase Cell Death Cell model Cells Cellular Stress Cerebral Amyloid Angiopathy Cerebral Edema Cerebrum Cessation of life Chronic Clinical Clinical Trials Data Dementia Deposition Development Disease Dose Drug usage Endothelial Cells Endothelium Energy-Generating Resources Enzymes Event Exhibits FDA approved Failure Family Functional disorder Future Glaucoma Goals Human Hydrogen Peroxide Immunohistochemistry Impaired cognition In Vitro Learning Link Mediating Membrane Potentials Memory Memory impairment Methazolamide Mitochondria Modeling Mus Nerve Degeneration Neurofibrillary Tangles Neuroglia Neurons Pathologic Pathology Pathway interactions Pharmacological Treatment Prevention Production Protons Reactive Oxygen Species Reporting Research Respiration Seizures Senile Plaques Synapses Tauopathies Testing Therapeutic Time Toxic effect Transgenic Mice Transgenic Model Translating analog behavioral phenotyping brain cell carbonate dehydratase cerebral amyloidosis cognitive function cytochrome c disorder control drug repurposing experimental study in vitro testing in vivo mitochondrial dysfunction mitochondrial membrane mouse model neurofibrillary tangle formation neurovascular novel novel therapeutics preservation prevent protective effect tau Proteins

项目摘要

Project Summary Mitochondria are considered one of the early targets in Alzheimer’s disease (AD). Preserving mitochondrial function can be a key strategy to prevent energetic failure in the AD brain and to inhibit the production of reactive oxygen species (ROS) and activation of caspases, thereby reducing neuronal, glial and vascular cell dysfunction and death, and hindering the progression of AD pathology. Our previous studies and others demonstrated that amyloid beta (Aβ) causes brain cell death through mitochondrial dysfunction and release of cytochrome C (CytC). Our preliminary data revealed that methazolamide (MTZ) and acetazolamide (ATZ), two carbonic anhydrase inhibitors (CAIs) highly active on mitochondrial carbonic anhydrases (CAs) are able to prevent mitochondrial dysfunction and CytC release in brain vascular and neuronal cells and in animal models of cerebral amyloidosis, through the inhibition of mitochondrial H2O2 production and mitochondrial depolarization. CAIs such as MTZ and ATZ are currently FDA approved for glaucoma, high altitude sickness, seizures, and other clinical indications. Previous studies reported increases in CAs expression in the aging and AD brain, confirming the relevance of CAs as a novel and previously unexplored target for AD therapy. We hypothesize that CA inhibition prevents mitochondrial dysfunction and cell death mechanisms induced by both Aβ and tau in vitro and in vivo, ameliorating neurovascular pathology and cognitive impairment in AD models. In Aim 1, we will perform a pharmacological treatment with the CAIs in a transgenic mouse model presenting both amyloidosis and tauopathy. We will feed 3xTg-AD transgenic mice with the two CAIs. We will start treating mice prior to onset of pathology (from 6 months of age), and when pathology is already present (from 11 months of age). Mitochondrial function, ROS production, caspase activation, cell death, amyloid and tau deposition will be evaluated in the mouse brain biochemically and by immunohistochemistry. Cognitive function will be evaluated behaviorally. In Aim 2, we will assess in vitro the efficacy of CAIs for prevention of tau-mediated mitochondrial dysfunction, using human neuronal, glial and endothelial cells in culture challenged with phosphorylated (pathological) tau species. In Aim 3, we will validate mitochondrial carbonic anhydrases as important targets associated to AD pathology and susceptible to modulation by amyloid and tau challenge. We will assess expression levels and specific localization of CA-VA, -VB and -II in age-matched AD and healthy human brains. We will silence mitochondrial CAs in endothelial, glial and neuronal cells to test the hypothesis that CA silencing will prevent mitochondrial dysfunction pathways and cell death induced by Aβ and tau pathologic species similarly to CA inhibition. Goal of the proposed line of research is to validate CAs as a target for amyloidosis and tauopathy and translate this study to a clinical trial to repurpose CAIs for AD and other dementias in the close future.

项目概要线粒体被认为是阿尔茨海默病 (AD) 的早期目标之一。功能可能是防止 AD 大脑能量衰竭和抑制产生的关键策略活性氧 (ROS) 和半胱天冬酶的激活，从而减少神经、神经胶质和血管细胞的活性功能障碍和死亡，并阻碍 AD 病理学的进展。 β 淀粉样蛋白 (Aβ) 通过线粒体功能障碍和释放我们的初步数据显示，醋甲唑胺 (MTZ) 和乙酰唑胺 (ATZ) 是两种细胞色素 C (CytC)。对线粒体碳酸酐酶 (CA) 高度活跃的碳酸酐酶抑制剂 (CAI) 能够预防脑血管和神经元细胞以及动物模型中的线粒体功能障碍和 CytC 释放通过抑制线粒体 H2O2 的产生和线粒体来治疗脑淀粉样变性 MTZ 和 ATZ 等 CAI 目前已获 FDA 批准用于治疗青光眼、高原反应、癫痫发作和其他临床迹象表明，随着年龄的增长，CAs 的表达增加。 AD 大脑，证实了 CA 作为 AD 治疗的新型且先前未探索的靶点的相关性。我们认为 CA 抑制可以防止线粒体功能障碍和细胞死亡机制在体外和体内均由 Aβ 和 tau 诱导，改善神经血管病理学和认知能力在目标 1 中，我们将使用 CAI 进行逻辑药物治疗。呈现淀粉样变性和 tau 蛋白病的转基因小鼠模型我们将饲喂 3xTg-AD 转基因小鼠。我们将在小鼠发病前（从 6 个月大）开始治疗具有两种 CAI 的小鼠，以及当病理已经存在时（从 11 个月大起）。小鼠大脑中的激活、细胞死亡、淀粉样蛋白和 tau 沉积将通过生化方法进行评估在目标 2 中，我们将在体外评估认知功能。使用人类神经元、胶质细胞，CAI 预防 tau 介导的线粒体功能障碍的功效在目标 3 中，我们将用磷酸化（病理性）tau 蛋白攻击培养物中的内皮细胞。验证线粒体碳酸酐酶是与 AD 病理学相关的重要靶标我们将评估表达水平和特异性对淀粉样蛋白和 tau 蛋白调节的敏感性。 CA-VA、-VB 和 -II 在年龄匹配的 AD 和健康人脑中的定位我们将沉默线粒体。内皮细胞、神经胶质细胞和神经元细胞中的 CA，以检验 CA 沉默会阻止线粒体的假设 Aβ 和 tau 病理物种诱导的功能障碍途径和细胞死亡与 CA 抑制类似。拟议研究的目标是验证 CA 作为淀粉样变性和 tau 蛋白病的靶点，并转化为这项研究是一项临床试验，旨在在不久的将来重新利用 CAI 治疗 AD 和其他痴呆症。