Utility of Human Organoids for Safety and Efficiency Evaluations of Genome Editing Therapeutics

项目概述本申请中提出的研究将推进人体细胞在3D器官模型中进行体细胞基因组编辑的体外安全性和有效性测试。对于基因组编辑，人类有机化合物有可能成为一种理想的工具，因为治疗的目标是人类基因组，而人类基因组不能在任何其他物种中复制。有机化合物在产量和人类副作用的可预测性方面也有优势。然而，重要的是要在这种情况下测试有机类化合物的实用性和价值，以证明这是一种使研究新药成为可能的技术。为了实现这一目标，我们将生产一组不同的有机体，代表人类的肾、肝、脑、肺、视网膜和/或心脏，作为基因编辑应用程序非常感兴趣的重要器官系统。对于我们“盘中的身体”中的每一个器官谱系，我们将演示衡量剪辑率和副作用的测试。将对这些分析进行优化，以建立具有分析稳定性、重复性和分析范围的可量化测量的参考标准。有机数据集将与使用类似基因编辑技术的合作团队在并行工作中产生的数据集进行比较。其目的是展示在人类器官培养中的安全性和有效性分析，以及在其他系统中的互补评估，作为一种易处理的范例，以支持将基因组编辑疗法推进到人类临床试验。为了最大限度地发挥影响，我们将重点放在广泛适用于多器官系统中的各种基因组编辑疗法的分析上。来源于人类多能干细胞的有机类化合物具有许多关键的组织特征，包括复杂排列的多种细胞类型，并表达与稀有人群相关的特定疾病表型。出于监管方面的考虑，迫切需要确定它们的保真度和预测能力。在这些研究中，我们将展示人类器官与其他临床前模型之间的一致性和协同性。因此，提出的具体目标是通过评估人体器官中不良事件的剂量依赖效率来降低治疗性基因组编辑方法的风险。总的来说，这些研究将产生在人类器官中进行基因组编辑的模型，其结果可以与同源模型进行比较，以建立一个可以应用于一系列组织和疾病的调节范式。

PROJECT SUMMARY The studies proposed in this application will advance in vitro safety and efficiency testing for somatic cell genome editing in human cells in 3D organoid models. For genome editing, human organoids have the potential to be an ideal tool, as the therapeutic target is the human genome, which cannot be replicated in any other species. Organoids also have advantages for throughput and predictivity of human side effects. It is important, however, to test the utility and value of organoids in this context, for this to be demonstrated as an enabling technology for investigational new drugs. To achieve this, we will produce a set of diverse organoids representing human kidney, liver, brain, lung, retina, and/or heart as vital organ systems of great interest to gene editing applications. For each organ lineage in our 'body in a dish', we will demonstrate assays to measure editing rates as well as side effects. These assays will be optimized to establish reference standards with quantifiable measurements of assay stability, reproducibility, and analytical range. Organoid datasets will be compared with datasets produced in parallel efforts by collaborating teams using similar gene editing technologies. The objective is to demonstrate safety and efficiency assays in human organoid cultures in conjunction with complementary assessments in other systems as a tractable paradigm to support the advancement of genome editing therapeutics to human clinical trials. To maximize impact, we will focus on assays that will be broadly useful for a wide variety of genome editing therapeutics, in multiple organ systems. Organoids derived from human pluripotent stem cells possess many key features of tissues, including diverse cell types in sophisticated arrangements, and express specific disease phenotypes associated with rare populations. For regulatory consideration, there is a critical need to determine their fidelity and prediction capacity. In these studies, we will demonstrate concordance and synergy between human organoids and other preclinical models. Thus, the Specific Aim proposed is to de-risk therapeutic genome editing approaches by assessing dose-dependent efficiency with adverse events in human organoids. Collectively, these studies will produce models of genome editing in human organoids with outcomes that can be compared to orthologous models to establish a regulatory paradigm which can be applied to a range of tissues and diseases.