Machine Learning Augmented Discovery of AAV Capsids for Cell Type Specific Access into Human Neurons and Glia

项目摘要 人类大脑包含了令人惊讶的多样化的神经元和神经胶质细胞类型，分布在数十个 功能区。由大脑倡议细胞普查产生的单细胞基因组资源提供了 丰富的关于这种多样性的参考知识，包括存在于人类和非人类灵长类动物的细胞类型 大脑。然而，我们目前缺乏可扩展的技术来从基因上获取这些特定类型的细胞，这将 有必要促进我们对其功能的理解。 通过利用我们在病毒工程、单细胞分析、机器学习和非人类和 人类灵长类神经科学，我们建议开发一个可扩展的工具包，用于访问 人脑。具体地说，我们的U01应用程序将为细胞开发新型腺相关病毒(AAV)载体 使用机器学习的特定类型的基因传递增强衣壳文库的工程。在这笔试点拨款中， 我们寻求展示我们的方法的潜力，以支持制定一个全面的 脑细胞类型的设施，将使功能和翻译研究在广泛的范围内 的应用程序。 特别是，我们将应用这种方法来开发人类前脑细胞类型的AAV载体， 包括海马体。我们的工具将在使用分子的特异性验证后公开传播 和生理指标，并映射到来自相同大脑区域的现有单细胞基因组学资源 由大脑倡议细胞普查项目产生。 如果成功，我们的项目将建立一个可扩展的基础设施，用于在 原理非人类或人类灵长类动物大脑中的任何组织或细胞类型。

Project Summary The human brain contains an astonishing diversity of neuronal and glial cell types distributed across dozens of functional areas. Single cell genomics resources generated by the BRAIN Initiative Cell Census have offered a wealth of reference knowledge about this diversity, including cell types present in human and non-human primate brain. However, we currently lack scalable technologies to genetically access these specific cell types, which will be necessary to advance our understanding of their function. By leveraging our expertise in viral engineering, single cell analysis, machine learning, and non-human and human primate neuroscience, we propose to develop a scalable toolkit for accessing distinct cell types in the human brain. Specifically, our U01 application will develop novel adeno-associated viral (AAV) vectors for cell type-specific gene delivery using machine learning augmented engineering of capsid libraries. In this pilot grant, we seek to demonstrate the potential of our approach to support the development of a comprehensive Armamentarium for brain cell types that would enable functional and translational studies across a broad range of applications. In particular, we will apply this methodology to develop AAV vectors for cell types in the human forebrain, including the hippocampus. Our tools will be openly disseminated after validation for specificity using molecular and physiological metrics, and mapped onto existing single cell genomics resources from the same brain regions generated by the BRAIN Initiative Cell Census projects. If successful, our project will establish a scalable infrastructure for addressing and functionally investigating in principle any tissue or cell type in the non-human or human primate brain.