A consortium effort to translate therapies for neurological diseases via an ex vivo organotypic platform

项目概要/摘要 大脑疾病仍然是一个巨大的医学挑战。新平台允许在 迫切需要结合患者组织和高通量容量的高保真系统来桥接 基于细胞的检测和全动物检测之间的差距。我们建议多机构共同努力， 使用器官型脑切片培养物开发和启动治疗性神经药物的翻译 （OBSC）平台。我们的OBSC模型利用活脑切片中现有的细胞和细胞外环境， 可以对脑组织进行快速的功能测试我们的多学科团队，其中跨越三个CTSA计划 hubs（密歇根大学、杜克大学和佛罗里达大学）开发了多种神经系统疾病模型，并显示了 OBSC平台发现疾病新机制和确定新治疗方法的有效性 化合物.我们开发了OBSC建模的技术创新，包括提高吞吐量 脑切片技术、基于病毒和生物射弹的疾病相关基因转导技术、靶基因 敲除、使用报告基因测定的实时监测和新鲜患者组织的掺入。我们现在寻求 利用OBSC平台的力量和我们团队的经验，扩大OBSC的效用 平台，并最终改善对患有脑部疾病的患者的护理。我们的多机构团队 将汇集和分享新的疾病模型，治疗药物的策划小组，独特的分子工具， 试剂盒和临床患者组织。在神经退行性疾病、脑癌和缺血性疾病领域， 疾病，我们将在平台上增加疾病相关的功能，筛选治疗药物，并使新的 基于免疫的方法，临床研究发展最快的领域，特别关注大脑 微环境实现我们目标的方法是：（i）我们将使用神经退行性变的OBSC模型 疾病发展的方法，用于跟踪细胞信号，使用实时光学成像，定义分子 研究人员正在研究介导疾病的炎症驱动因素的途径，并确定有效的新治疗剂。(ii)我们将 使用新鲜的患者活检样本和侵袭性脑癌的OBSC模型来表征细胞和 脑肿瘤的基因组异质性，抗癌免疫疗法的疗效，并进行药物筛选 来鉴定新的治疗药物。(iii)最后，我们将利用缺血性疾病的OBSC模型来开发 用于合并分子筛选的方法，研究小胶质细胞对卒中进展的影响，并鉴定 减少梗塞面积的新化合物。总之，我们的方法将创建一个可扩展的基础设施， 围绕OBSC技术，加速发现新的和有效的治疗策略，并启动 转化为最终的人类患者试验，以治疗多种大脑疾病。

Project Summary/Abstract Disorders of the brain remain an enormous medical challenge. New platforms that allow functional testing in high-fidelity systems that incorporate patient tissue and high-throughput capacity are urgently needed to bridge gap between cell-based assays and whole-animal testing. We propose a multi-institutional effort to identify, develop, and initiate translation of therapeutic neurological agents using the organotypic brain slice culture (OBSC) platform. Our OBSC models leverage existing cellular and extracellular milieu in the live brain slices to allow rapid, functional testing on brain tissue. Our multi-disciplinary team, which spans three CTSA Program hubs (UNC, Duke, and Univ. Florida), has developed multiple models for neurological disorders and shown the effectiveness of the OBSC platform to discover new mechanisms of disease and identify new therapeutic compounds. We have developed technological innovations for OBSC modeling, including elevated-throughput techniques for brain slicing, viral- and biolistic-based transduction of disease-relevant genes, targeted gene knockdown, real-time monitoring using reporter assays, and incorporation of fresh patient tissue. We now seek to leverage the power of the OBSC platform and experience of our team to broaden the utility of the OBSC platform and ultimately improve the care for patients suffering from brain disorders. Our multi-institutional team will bring together and share new disease models, curated panels of therapeutic agents, unique molecular tool kits, and clinical patient tissue. Within the areas of neurodegenerative disease, brain cancers and ischemic disease, we will add disease-relevant capabilities to the platform, screen therapeutic agents, and enable new immune-based approaches, the fastest-growing area of clinical research, with a particular focus on the brain microenvironment. Approaches to accomplish our aims are: (i) We will use OBSC models of neurodegenerative disease to develop methods for tracking of cell signaling using real-time optical imaging, define molecular pathways mediating inflammatory drivers of disease, and identify effective new therapeutic agents. (ii) We will use fresh patient biopsy samples and OBSC models of aggressive brain cancer to characterize he cellular and genomic heterogeneity of brain tumors, the efficacy of anti-cancer immune therapies, and perform drug screens to identify new therapeutic agents. (iii) Lastly, we will utilize OBSC models of ischemic disease to develop methods for pooled molecular screening, investigate the impact of microglia on stroke progression, and identify new compounds that reduce infarct size. Together, our approaches will create an expandable infrastructure built around OBSC technology, accelerate the discovery of new and effective therapeutic strategies, and initiate translation towards ultimate human patient trials to treat multiple disorders of the brain.