The CRISPR Vision Program: Nonviral Genome Editing Platforms to Treat Inherited Retinal Channelopathies

项目总结/摘要-总体 基因组编辑社区正在庆祝2022年具有里程碑意义的CRISPR论文发表十周年。 对该领域的许多人来说，值得注意的是编辑精度和多功能性方面的巨大技术进步， 沿着而来的是诺贝尔奖和数十亿美元的投资。然而，有一种明显的感觉， 这些进展没有以足够的速度转化为有价值的药物。体内基因编辑仍面临 这是一个巨大的挑战，特别是在安全性，有效性和交付方面。尽管制订 EDIT-101是一种携带Cas9的病毒载体，用于治疗一种罕见的视网膜疾病（BRILLIANCE试验）， 对于基因组编辑治疗计划的诊所来说，这条道路对于其他人来说可能是具有挑战性的。 致力于开发人类疗法的领域。病毒策略有几个局限性，涉及免疫 对矢量元素的反应和编辑器的终身表达，提高 偏离目标的问题较新的编辑器，如基础编辑器，不能容易地包装到常见的病毒载体中， 病毒载体制造的供应链具有挑战性。我们试图克服病毒的局限性， 通过使用称为二氧化硅纳米胶囊（SNC）和靶向的新型非病毒递送系统， Active Gene Editors（TAGE）.它们可以高效地将基因组编辑器输送到视网膜， 达到10-70%的水平，这是最好的非病毒交付编辑的眼睛和可比的， 病毒输送系统。基于我们强有力的已发表和初步数据，我们建议开发非病毒基因， 编辑产品，以治疗最好的疾病（BD）和利伯先天性黑蒙（LCA），两种疾病影响 视网膜色素上皮的离子通道（即，RPE通道病）。我们的团队跨越学术界和 工业，将追求以下目标。在总体目标1中，我们评估了基因组编辑器的临床准备情况 通过瞬时、局部和非病毒递送靶向有丝分裂后细胞。到目前为止，在肝脏之外，只有 病毒编辑器已经达到了用于体内编辑的IND。在这里，我们严格评估SNC的潜力， 用于治疗LCA和BD的TAGE非病毒递送系统。我们将最大的努力投入到 领导项目1，在SNC中开发基本编辑器。该项目将在五年内达到IND， 为其他项目提供协同效应。在总体目标2中，我们创建了一个可以解决许多罕见疾病的平台 通过使用简单的临床前测试流水线生产不同的导向器，整体 目标3，我们为SCGE联盟和更广泛的基因组编辑领域提供了一套调控相互作用， 阐明了新基因编辑疗法的临床发展路径。最后，通过我们在BD的经验， 我们希望了解在开发体细胞基因组编辑器的情况下， 存在合适的动物模型。因为大多数已知的眼部病理性突变没有合适的 动物模型，分享这些知识将对随后的基因组编辑线索产生很大影响。

PROJECT SUMMARY/ABSTRACT – OVERALL The genome editing community is celebrating the ten-year anniversary of landmark CRISPR papers in 2022. What is notable to many in the field is tremendous technological advances in editing precision and versatility, along with a Nobel Prize and billions of dollars of investment. However, there is a palpable sense that these advances have not been translated into valuable drugs at a sufficient rate. In vivo gene editing still faces substantial challenges, especially when it comes to safety, efficacy, and delivery. While the development of EDIT-101, a viral vector carrying Cas9 to treat a rare retinal disorder (BRILLIANCE trial), provided a clear path to the clinic for a genome editing therapeutic program, this path can be challenging to follow for others in the field interested in developing human therapeutics. Viral strategies have several limitations involving an immune response to vector elements and prolonged expression of the editor for the lifetime of the patient, heightening off-target concerns. Newer editors like base editors cannot be readily packaged into common viral vectors, plus there is a challenging supply chain for viral vector manufacturing. We seek to overcome the limitations with viral delivery systems by using novel nonviral delivery systems termed the Silica NanoCapsule (SNC) and Target Active Gene Editors (TAGE). They can efficiently deliver genome editors to the retina with high efficiency, reaching levels of 10-70% that are among the best for nonviral delivery of editors to the eye and comparable to viral delivery systems. Based on our strong published and preliminary data we propose to develop nonviral gene editing products to treat Best Disease (BD) and Leber Congenital Amaurosis (LCA), two diseases affecting ion channels of the retinal pigment epithelium (i.e., RPE channelopathies). Our team, spanning academia and industry, will pursue the following aims. In Overall Aim 1, we evaluate the clinical readiness of a genome editor targeting a post-mitotic cell through transient, localized, and nonviral delivery. To date, outside of the liver, only viral editors have reached an IND for in vivo editing. Here, we rigorously evaluate the potential of our SNC and TAGE nonviral delivery systems for the treatment of LCA and BD. We invest our most significant effort into the Lead Project 1 that develops a base editor within a SNC. This project will reach an IND within five years and provide synergy for other projects. In Overall Aim 2, we create a platform that can address many rare diseases of the eye by streamlined manufacturing of different guides with a simple pipeline for preclinical testing. In Overall Aim 3, we provide the SCGE Consortium and broader genome editing field a set of regulatory interactions that clarifies development path to the clinic for new gene editing therapies. Finally, through our experience with BD, we expect to learn about the regulatory path in developing somatic cell genome editors for scenarios when no suitable animal model exists. Because the majority of known pathological mutations in the eye have no suitable animal models, sharing this knowledge will have a large impact on subsequent genome editing leads.