Prospective Evaluation of Chloride Channel-Targeted Therapy for Alzheimer's disease

摘要 本申请是为了响应特别利益通知（NOSI）而提交的，该通知被标识为NOT-AG- 22-025.阿尔茨海默病（Alzheimer's disease，AD）是最常见的痴呆症，也是导致残疾的主要原因之一 和老年人的死亡。它对社会造成了重大的经济影响。临床AD的患病率 11.3%（美国610万人），预计到2060年这一数字将上升到1400万。因此，治疗 管理或延迟AD的过程可以显着减少日益增加的医疗负担。但我们 经过40年的研发努力，仍然缺乏有效的干预措施。FDA的第一个疾病修饰淀粉样蛋白 靶向治疗Aduhelm减少了患者大脑中的淀粉样蛋白沉积，但尚未显示出影响 临床结果，如认知能力下降的进展。这表明，迫切需要 就像我们有很多治疗癌症的方法一样，cGAS-sting 信号传导被认为是神经病理生理学的关键决定因素，因为在 阿尔茨海默病或相关痴呆症。然而，目前没有可用的cGAS或STING抑制剂 在临床阶段。目前正在开发具有良好药物特性的临床可行的抑制剂。 在我们的母公司奖中，亚细胞氯报告基因和细胞器化学型指纹 技术可以帮助我们研究细胞器氯的生理作用，这是研究细胞器氯的关键。 开发研究不足的氯离子通道靶向治疗。在我们的初步研究中，我们使用了非- 选择性氯离子通道阻断剂诱导全细胞氯离子失调。我们发现非- 选择性氯离子通道阻断剂抑制cGAS-STING信号传导的剂量- 依赖性方式而不引起细胞死亡。我们假设细胞/细胞器氯离子起着 在cGAS-STING信号通路中的重要作用。阿尔茨海默病中cGAS-STING信号的升高 通过氯离子通道靶向干扰氯离子水平可以抑制疾病。我们提出了一个 一项前瞻性研究，旨在评估氯离子通道靶向治疗阿尔茨海默病的合格性， cGAS-STING途径的衰减。在目标1中，我们将绘制cGAS-STING期间的氯稳态图。 激活和阿尔茨海默病。目的2重点研究氯离子在cGAS-STING中的作用 途径，并确定氯离子通道靶向治疗抑制cGAS-STING升高的能力。 阿尔茨海默病的发病机制拟议的研究整合了细胞器氯化物成像，氯化物 生理学研究（cGAS-STING）和氯离子通道靶向治疗阿尔茨海默病。在 在拟议的研究结束时，我们预计将了解细胞氯在cGAS-STING途径中的作用 并前瞻性评估氯离子通道靶向治疗抑制cGAS升高的资格， STING信号在阿尔茨海默病中的作用为氯生理学研究奠定了基础， 阿尔茨海默病和相关神经系统疾病的氯离子通道靶向治疗的验证。

ABSTRACT This application is being submitted in response to the Notice of Special Interest (NOSI) identified as NOT-AG- 22-025. As the most common form of dementia, Alzheimer's disease (AD) is one of the major causes of disability and death among older people. It causes significant economic impacts on society. The prevalence of clinical AD was 11.3% (6.1 million people in US) and this number is predicted to rise to 14 million in 2060. Therefore, therapy that manages or delays AD’s process could significantly reduce the increasing healthcare burden. However, we still lack effective interventions for AD after 40 years of R&D efforts. The FDA’s first disease-modifying, amyloid- targeted therapy, Aduhelm, reduces amyloid deposits in the patient's brain but it has not yet been shown to affect clinical outcomes such as progression of cognitive decline. It indicates that there is an urgent need to have more options for treating Alzheimer’s disease, just as we have many treatments for cancer. cGAS-STING signaling is recognized as a crucial determinant of neuropathophysiology as the elevated signal is observed in Alzheimer's disease or related dementia. However, there are currently no cGAS or STING inhibitors available in clinical stage. Work is underway to develop clinically viable inhibitors with good drug properties. In our parent award, the subcellular chloride reporters and the organellar chemotype fingerprinting techniques could help us to investigate the physiological role of organellar chloride, which is the key to the development of under-studied, chloride channel-targeted therapy. In our preliminary study, we used non- selective chloride channel blockers to induce the whole cell chloride dysregulation. We found that the non- selective chloride channel blockers inhibited the cytosolic-DNA stimulated cGAS-STING signaling in a dose- dependent manner without causing cell death. We hypothesize that cellular/organellar chloride plays an important role in the cGAS-STING signaling pathway. The elevated cGAS-STING signaling in Alzheimer's disease could be inhibited by disturbing the chloride levels via chloride channel targeting. We propose a prospective study, to evaluate the eligibility of chloride channel-targeted therapy for Alzheimer's disease via the attenuation of cGAS-STING pathway. In Aim 1, we will map the chloride homeostasis during the cGAS-STING activation and in Alzheimer's disease. Aim 2 focuses on investigating the role of chloride in cGAS-STING pathway and determine the ability of chloride channel-targeted therapy to suppress the elevated cGAS-STING pathway in Alzheimer's disease. The proposed research integrates organellar chloride imaging, chloride physiology investigation (cGAS-STING), and chloride channel-targeted therapy for Alzheimer’s disease. At the end of the proposed study, we anticipate understanding the role of cellular chloride in cGAS-STING pathway and prospectively evaluate the eligibility of chloride channel-targeted therapy to dampen the elevated cGAS- STING signaling in Alzheimer's disease. It will lay the foundation for chloride physiology and prospective validation of chloride channel-targeted therapy for Alzheimer's disease and related neurological disorders.