Construction of an Integrated Immune - Vascular Brain - Chip as a Platform for the Study, Drug Screening, and Treatments of Alzheimer's Disease

摘要 阿尔茨海默病(AD)是一种使人衰弱的大脑疾病，其人力和经济成本都令人震惊。而当 遗传学研究越来越多地发现与阿尔茨海默病相关的多态，但仍没有清晰的图景 分子和细胞参与者以及每个参与者对AD的贡献程度。基因和 阿尔茨海默病的分子复杂性以及缺乏在人体组织中实验性地分解它的技术 制约疗法发现及其成功应用于临床的瓶颈。vbl.使用 我们最近开发了一种体外血脑屏障(IBBB)，并将其应用于发现 导致脑淀粉样血管病(CAA)遗传易感性的机制。与临床研究相同， 我们发现，CAA和AD的最强遗传风险因素APOE4显著增加了淀粉样蛋白 在我们的iBBB中的证词。我们的工程组织的易操纵性使我们能够剖析细胞原因。 这种疾病的危害。我们发现仅在周细胞中APOE4的表达就足以增加脑血管 淀粉样蛋白堆积。精确定位介导CAA风险的因果细胞然后使分子和 建立了潜在机制并揭示了新的治疗机会的生化研究 降低CAA和潜在AD的遗传风险。在这里，我们将在成功的基础上，使用iBBB作为 支架；我们将整合神经元、少突胶质细胞和小胶质细胞，以在其上生成微整合的大脑 芯片(miBrain-Chip)。在UG3 Aim1.1中，我们将建立代表健康和疾病的miBrain芯片 通过包含最先进的生物聚合物的迭代优化来了解人脑的状态 以及麻省理工学院罗伯特·兰格实验室的工程专业知识。UG3 Aim1.2将从基因上进行整合和验证 神经元活动的编码调制器和报告器使miBrain芯片能够研究神经元如何 活动受到影响，进而影响AD的发病机制。UG3 AIM2将模拟病理 阿尔茨海默病在男性和女性SAID IPSC系队列中的AD进展 匹配的大脑样本、临床病史和基因组序列。我们将建立计算模型 描述了导致终态的转录、细胞动力学和组织学转变 死后AD的大脑。这些纵向病理图谱来自不同基因的健康和悲伤 个人将获得对AD发展的机械性洞察，并创建一个发现和功效的平台 治疗药物的筛选。我们假设AD的潜在机制受到以下因素的显著影响 遗传变异性。在UH3中，我们将建立APOE4致病机制(UH3Aim1)和 然后确定一组临床前和临床AD药物的疗效、毒性和治疗窗口 使用等基因APOE3和APOE4miBrain芯片(UH3AIM2)。我们的多式联运战略将阐明如何 基因变异影响AD的发病机制和治疗反应，开辟了治疗AD的新途径 迅速发现药物并将有效的治疗方法转化为临床。

Abstract Alzheimer's disease (AD) is a debilitating brain disorder, with staggering human and financial cost. While genetic studies are increasingly identifying polymorphisms that correlate with AD, there still is no clear picture of the molecular and cellular players and the extent to which each contributes to AD. The genetic and molecular complexity of AD and the lack of technology for experimentally unraveling it in human tissues create a bottleneck constricting the discovery of therapeutics and their successful translation into the clinic. Using human iPSCs we recently developed an in vitro blood-brain barrier (iBBB) and deployed it to discover mechanisms causing genetic predisposition to cerebral amyloid angiopathy (CAA). Identical to clinical studies, we found that APOE4, the strongest genetic risk factor for CAA and AD significantly increased amyloid deposition in our iBBB. The tractability of our engineered tissues then enabled dissection of the cellular causes of the disease. We found expression of APOE4 in pericytes alone was sufficient to increase cerebral vascular amyloid accumulation. Pinpointing the causal cells mediating CAA risk then enabled molecular and biochemical studies that established the underlying mechanism and revealed new therapeutic opportunities for mitigating genetic risk of CAA and potentially AD. Here, we will build upon our success, using the iBBB as a scaffold; we will incorporate neurons, oligodendrocytes, and microglia to generate a micro-integrated brain on a chip (miBrain-chip). In UG3 Aim1.1 we will establish miBrain-chips that represent healthy and diseased states of the human brain through iterative rounds of optimization that incorporate state-of-the-art biopolymers and engineering expertise from Robert Langer's lab at MIT. UG3 Aim1.2 will integrate and validate genetically encoded modulators and reporters of neuronal activity enabling the miBrain-chip to investigate how neuronal activity is influenced, and in turn, influences AD pathogenesis. UG3 Aim2 will model the pathological progression of AD in miBrain-chips across cohort of male and female sAD iPSC lines for which we have matched brains samples, clinical history, and genomic sequences. We will build computational models describing the transcriptional, cellular-dynamics and histological transformations that lead up to the end-states of post-mortem AD brains. These longitudinal pathological maps from genetically diverse healthy and sAD individuals will yield mechanistic insight into AD development and create a platform for discovery and efficacy screening of therapeutics. We hypothesize that the mechanisms underlying AD are significantly influenced by genetic variability. In UH3 we will establish the mechanisms underlying APOE4 pathogenesis (UH3 Aim1) and then ascertain the efficacy, toxicity, and therapeutic window of a panel of preclinical and clinical AD drugs using isogenic APOE3 and APOE4 miBrain-chips (UH3 Aim2). Our multimodal strategy will shed light on how genetic variation influences AD pathogenesis and therapeutic response, opening up new avenues for expeditious drug discovery and translation of effective therapeutics to the clinic.