A Single-cell Platform for Analyzing the Peripheral Immune Response in Alzheimer’s and Alzheimer’s Related Diseases

随着第二阶段的提交，Isoplexis将开发一个自动化系统来评估合并的外围设备 T细胞和单核细胞的免疫应答对神经退行性疾病的鉴别和监测 包括阿尔茨海默氏症(AD)、额颞性痴呆(FTD)和多发性硬化症(MS)。监测疾病 进展和治疗对免疫反应的影响目前需要危险的脑脊液 取样。然而，中枢和外周免疫系统之间的沟通是存在的，我们 表明它需要一个复杂的单细胞检测系统来破译血液样本中的这些相互作用。 目前ADRD治疗的有限成功导致了两个关键观察：(1)靶向分子 不影响炎症反应或疾病进展的疾病贡献者，或(2) 靶向免疫治疗不能提供可测量的生产性结果，并引发不利的炎症 副作用。多重蛋白生物标记物已被证明在其他人类免疫疗法(例如癌症)中至关重要， 极大地提高了临床成功率，并激励了这项技术向ADRD的扩展。发展中 ADRD的这种生物标记物需要一种技术，该技术(1)在单个- 根据ADRD外周血细胞亚群分类的细胞平台，(2)分离敏感的外周免疫细胞 有必要建立相关的生物标记物疾病概况，以及(3)能够监测这些新出现的 治疗后的生物标志物。Isoplexis IsoCode的多功能强度指数(PSI)，使用30种分泌物 每个T细胞的蛋白质，已被证明是多种类型的免疫疗法中客观反应的相关性。在……里面 一项针对多发性硬化症患者样本的公开研究表明，Isoplexis平台具有强大的刺激性和 单核细胞炎性细胞因子分泌谱，与健康供者比较，以及这种分泌 治疗成功后，侧貌会大大减少。对AD外周T细胞的初步研究表明 与年龄匹配的供者相比，由独特的多功能亚群驱动的炎性特征增加。使用一个 专注于ND市场需求，我们希望将这项技术转化为完全可行的商业， 自动检测系统，极大地加快了ADRD疗法的行业发展。本第二阶段计划 支持在具有强大科学表现历史的临床环境中进行Beta测试。目标1：开发自动化的 “Flow cell”消耗品隔间，可并行捕获ADRD单核细胞和T细胞，并测量32- Plex单细胞细胞因子分泌。目标2：生产小型化、台式自动化流动池分析和 工作流系统，带有细胞增菌模块，为临床实验室提供人性化的样本答疑格式。 目标3：在UCI、UCSF和UConn试验中演示使用Isoplexis自动化的概念验证能力，并 神经退行性疾病比较研究中外周免疫细胞的多功能反应。

With this Phase II submission, IsoPlexis will develop an automated system to evaluate the combined peripheral immune response of T cells and monocyte cells to differentiate and monitor neurodegenerative diseases (ND) including Alzheimer’s (AD), Frontotemporal Dementia (FTD), and Multiple Sclerosis (MS). Monitoring disease progression and treatment impact on the immune response currently requires risky Cerebral Spinal Fluid sampling. However, the communication between the central and peripheral immune systems exists and we show it requires a sophisticated single-cell detection system to decipher these interactions from a blood sample. The limited success of current ADRD treatments has led to two key observations: (1) targeting molecular contributors to disease without impacting the inflammatory response or disease progression, or (2) immunotherapy targeting not providing measurably productive outcomes and inducing unfavorable inflammatory side effects. Multiplexed protein biomarkers have proven critical in other human immune therapies (e.g., cancer), drastically improving clinical success and inspiring the extension of this technology towards ADRD. Developing such biomarkers for ADRD requires a technology that (1) measures secreted protein polyfunctionality in a single- cell platform classified by subsets of ADRD peripheral blood cells, (2) isolates sensitive peripheral immune cells necessary to establish correlative biomarker disease profiles, and (3) is able to monitor these emerging biomarkers upon treatment. The IsoPlexis IsoCode’s polyfunctional strength index (PSI), using 30+ secreted proteins per T-cell, has proven to be a correlate of objective response in multiple types of immunotherapies. In a published study on MS patient samples, the IsoPlexis platform has demonstrated a strong stimulatory and inflammatory cytokine secretion profile from monocytes, compared to that of healthy donors, and this secretion profile is greatly reduced upon successful treatment. Initial studies of AD peripheral T cells have shown an increased inflammatory profile relative to age-matched donors driven by unique polyfunctional subsets. With a unique focus on the ND market need, we wish to transition this technology into a fully viable commercial, automated assay system to greatly expedite the industry’s development of ADRD therapies. This Phase II plan enables beta-testing in clinical settings with strong scientific performance history. Aim 1: Develop an automated “flow cell” consumable compartment, which captures ADRD monocytes and T cells in parallel and measures 32- plex single-cell cytokine secretions. Aim 2: Produce a miniaturized, benchtop automated flow cell analysis and workflow system, with cell enrichment module, to provide clinical labs user-friendly sample to answer format. Aim 3: Demonstrate proof-of-concept ability to use IsoPlexis automation at UCI, UCSF and UCONN trials and to profile polyfunctional response of peripheral immune cells in a comparative study of neurodegenerative diseases.