3D patient-derived organoids: A viable and complementary model to study neural system dysfunction in Alzheimer's Disease

3D 患者来源的类器官：研究阿尔茨海默病神经系统功能障碍的可行且互补的模型

• 批准号: 2641001
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2641001
• 关键词: 3D; patient; derived; organoids; viable

Alzheimer's Disease (AD) is a devastating disorder for which effective disease-modifying treatments are yet to be identified. Recent evidence from our laboratory indicates that neural circuit dysfunction emerges very early in AD, preceding cognitive deficits and shaping subsequent disease progression, ultimately leading to neurodegeneration and dementia. Neural circuits in AD exhibit early neuronal hyperexcitability which transitions to hypoactivity in later disease, as a result of the pathological effects of Amyloid beta and tau (the hallmark proteins that accumulate in AD), respectively. Mouse models have been crucial to dissecting these circuit-level changes with high spatiotemporal resolution while providing remarkable insights into potential pathological mechanisms and candidate therapeutic targets. Nevertheless, clinical translation of promising therapies in preclinical models (e.g., reduction/removal of Amyloid beta) has so far proven unviable, highlighting the limitations of animal-models as well as the challenge in disambiguating human-disease relevant pathological alterations. Current research investigating changes in neural circuit function in AD is typically conducted in mice that carry mutations/transgenes or manifest proteinopathies which reproduce phenotypes observed in human disorders. Such experiments (often of moderate severity) are highly complex, invasive, and of low-throughput, as it is often necessary to perform surgeries and implant cranial apparatus and inject biosensors/implant electrodes to enable high-resolution in-vivo recordings. These challenges emphasise the need to identify and validate complimentary and alternative models that more faithfully recapitulate human brain circuits in AD and beneficially address the use of animal models in AD research.Technical advances in patient-derived induced pluripotent stem cell (iPSC) methodologies have recently emerged that provide an auxiliary approach with which to model human disease that can be studied and modulated under tightly controlled experimental conditions. Notably, our collaborator on this proposal, Prof. Selina Wray at UCL, building on work funded by the NC3Rs (through CRACK-IT in 2013 on the Untangle project, https://nc3rs.org.uk/crackit/untangle), has successfully generated long-term viable and homogeneous cerebral organoids from familial AD and tauopathy patients which form complex neuronal circuits and permit the study of disease-relevant circuit-level changes in human-associated neural components. Here, we propose to uniquely integrate this expertise with that of the Busche Laboratory's in state-of-art structural and functional recording techniques to study neuronal circuits in AD (i.e., Neuropixels multi-channel electrophysiology, two-photon calcium imaging, 3D lightsheet microscopy). In so doing, we aim to leverage and combine the cutting-edge methods established by both groups to establish a radically novel platform to gain transformative insights into AD circuit function, promote clinical translation, and significantly reduce reliance on animal models. Patient-derived iPSC cultures will have the complete disease-relevant genetic profile, and the proposed high-resolution recording techniques will permit the detailed investigation of disease mechanisms in relevant cell types and overlying circuits, and provide a novel opportunity to interrogate the early cellular and molecular changes that lead to the development of clinical AD. With the increasing recognition of the importance of early neural circuit dysfunction in AD, and a growing number of laboratories worldwide utilising animal models to understand the mechanistic underpinnings of neuronal circuitry in AD, we anticipate that validation of our organoid model will lead to a substantial replacement of 25% of existing animal work, not only in our laboratory but also, subsequently, in the ~20 laboratories worldwide (equating to ~300 animals/year) which study circuit dysfunction in AD.

阿尔茨海默病 (AD) 是一种毁灭性的疾病，目前尚未确定有效的疾病缓解治疗方法。我们实验室的最新证据表明，神经回路功能障碍在 AD 的早期就出现，先于认知缺陷并影响随后的疾病进展，最终导致神经变性和痴呆。 AD 中的神经回路表现出早期神经元过度兴奋，在后期疾病中会转变为活动减退，这分别是由于淀粉样蛋白 β 和 tau（AD 中积累的标志蛋白）的病理作用的结果。小鼠模型对于以高时空分辨率剖析这些电路水平的变化至关重要，同时提供对潜在病理机制和候选治疗靶点的深刻见解。然而，迄今为止，临床前模型中有前途的疗法（例如减少/去除β淀粉样蛋白）的临床转化已被证明是不可行的，这凸显了动物模型的局限性以及消除人类疾病相关病理改变的挑战。目前研究 AD 中神经回路功能变化的研究通常是在携带突变/转基因或表现出再现人类疾病中观察到的表型的蛋白质病的小鼠中进行的。此类实验（通常为中等严重程度）非常复杂、具有侵入性且通量低，因为通常需要进行手术和植入颅骨装置并注射生物传感器/植入电极以实现高分辨率体内记录。这些挑战强调需要识别和验证互补和替代模型，以更忠实地再现 AD 中的人类大脑回路，并有益地解决 AD 研究中动物模型的使用问题。最近出现的患者来源诱导多能干细胞 (iPSC) 方法的技术进步，为模拟人类疾病提供了一种辅助方法，可以在严格控制的实验条件下研究和调节。值得注意的是，我们这项提案的合作者，伦敦大学学院的 Selina Wray 教授，在 NC3Rs 资助的工作（2013 年通过 CRACK-IT 的 Untangle 项目，https://nc3rs.org.uk/crackit/untangle）的基础上，成功地从家族性 AD 和 tau 病患者中生成了长期可行且均质的大脑类器官，这些患者形成了复杂的神经元回路 并允许研究人类相关神经组件中与疾病相关的回路水平变化。在这里，我们建议将这一专业知识与 Busche 实验室最先进的结构和功能记录技术相结合，以研究 AD 中的神经元回路（即 Neuropixels 多通道电生理学、双光子钙成像、3D 光片显微镜）。在此过程中，我们的目标是利用和结合两个小组建立的尖端方法，建立一个全新的平台，以获得对 AD 回路功能的变革性见解，促进临床转化，并显着减少对动物模型的依赖。患者来源的 iPSC 培养物将具有完整的疾病相关遗传图谱，所提出的高分辨率记录技术将允许详细研究相关细胞类型和覆盖回路的疾病机制，并提供一个新的机会来探究导致临床 AD 发展的早期细胞和分子变化。随着人们越来越认识到 AD 早期神经回路功能障碍的重要性，以及世界各地越来越多的实验室利用动物模型来了解 AD 神经元回路的机制基础，我们预计，我们的类器官模型的验证将导致 25% 的现有动物工作的实质性替代，不仅在我们的实验室，而且随后在全球约 20 个实验室（相当于约 300 个实验室）中进行。 动物/年）研究 AD 中的回路功能障碍。