Walton Chaska C, Wang Ellen, Lee Suckwon, Siebrand Cynthia J, Bergo Nicholas J, Mayeri Zachary, Andersen Julie K
Buck Institute for Research on Aging, 8001 Redwood Blvd, Novato, CA, 94945, USA.
SENS Research Foundation, Mountain View, CA, 94041, USA.
Alzheimers Res Ther. 2025 Jul 26;17(1):176. doi: 10.1186/s13195-025-01822-7.
Therapeutic successes in mouse models of Alzheimer’s disease (AD) largely fail to translate into clinical trials, with experimental drugs rarely validated in human models before being administered to humans. To address this, we developed an accessible method to culture commercially available primary human neurons and astrocytes, along with an amyloid-beta 1–42 (Aβ)-based in vitro AD model. This system enables to reliably culture primary neurons to mature stages of development, essential to model the adult brain and neurodegenerative diseases such as AD. The absence of a blood–brain barrier (BBB) in this model permits evaluation of drug mechanisms of action independently of BBB permeability, thereby informing the feasibility of developing fully BBB-penetrant therapeutics for CNS interventions based on validated senolytic pathways. Using this platform, we evaluated two senolytic regimens previously shown to be effective in AD mouse models: Navitoclax (NAV), which targets the Bcl-2 family of anti-apoptotic proteins, and the dasatinib–quercetin (DQ) cocktail, which inhibits tyrosine kinases and AKT signaling, among other pathways. We also assess the natural killer cell line NK92 to model emerging immune-mediated senescent cell ablation therapies. In synaptically mature cultures, we show that NK92 cells preferentially—but not exclusively—targeted Aβ-treated neurons and astrocytes with senescent-like phenotypes. DQ demonstrated a safe profile for human neurons, but Navitoclax exhibited non-selective neurotoxicity. These findings highlight potential risks associated with developing BBB-permeable therapies based on the mechanisms of NAV and NK cell-mediated cytotoxicity. Our work underscores the critical need for human-relevant models in the AD drug-development pipeline to improve safety and clinical translatability.
The online version contains supplementary material available at 10.1186/s13195-025-01822-7.
阿尔茨海默病（AD）小鼠模型的治疗成功在很大程度上未能转化为临床试验，实验药物在用于人体之前很少在人体模型中得到验证。为了解决这个问题，我们开发了一种可获取的方法来培养市售的原代人神经元和星形胶质细胞，以及一种基于β淀粉样蛋白1-42（Aβ）的体外AD模型。该系统能够可靠地将原代神经元培养至发育成熟阶段，这对于模拟成人大脑和诸如AD等神经退行性疾病至关重要。该模型中不存在血脑屏障（BBB），允许独立于BBB通透性评估药物作用机制，从而为基于已验证的衰老细胞溶解途径开发用于中枢神经系统干预的完全BBB穿透性治疗药物的可行性提供信息。使用这个平台，我们评估了之前在AD小鼠模型中显示有效的两种衰老细胞溶解方案：靶向抗凋亡蛋白Bcl-2家族的Navitoclax（NAV），以及抑制酪氨酸激酶和AKT信号传导等途径的达沙替尼-槲皮素（DQ）鸡尾酒。我们还评估了自然杀伤细胞系NK92，以模拟新兴的免疫介导的衰老细胞消融疗法。在突触成熟的培养物中，我们表明NK92细胞优先但非排他性地靶向具有衰老样表型的Aβ处理的神经元和星形胶质细胞。DQ对人神经元显示出安全的特征，但Navitoclax表现出非选择性神经毒性。这些发现突出了基于NAV机制和NK细胞介导的细胞毒性开发BBB穿透性疗法的潜在风险。我们的工作强调了在AD药物开发流程中建立与人类相关模型以提高安全性和临床可转化性的迫切需求。
在线版本包含可在10.1186/s13195-025-01822-7获取的补充材料。
