Silica Nanocapsule-Mediated Nonviral Delivery of CRISPR Base Editor mRNA and Allele Specific sgRNA for Gene Correction in Leber Congenital Amaurosis

二氧化硅纳米胶囊介导的 CRISPR 碱基编辑器 mRNA 和等位基因特异性 sgRNA 非病毒传递用于 Leber 先天性黑蒙的基因校正

• 批准号: 10668166
• 负责人: SHAOQIN GONG
• 金额: $ 186.61万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-16 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10668166
• 关键词: 3-Dimensional; Acceleration; Alleles; Animals; Biological Products; Biomanufacturing; Blindness; Cells; Cellular Assay; Chemicals; Chemistry; Childhood; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Data; Development; Dose; Drug Packaging; Engineering; Eye; FDA approved; Formulation; Future; Genes; Good Manufacturing Process; Guide RNA; Human; Immune; In Vitro; Inherited; Investigational Drugs; Investigational New Drug Application; Kir7.1 channel; Knowledge; Lead; Leber' s amaurosis; Ligands; Mediating; Messenger RNA; Methods; Modality; Mus; Particle Size; Patients; Pharmaceutical Preparations; Point Mutation; Prevalence; Process; Production; Property; Quality Control; Rattus; Research Contracts; Research Methodology; Retina; Retinal Diseases; Running; Safety; Silicon Dioxide; Specificity; Structure of retinal pigment epithelium; Surface; Technology Transfer; Testing; Therapeutic Effect; Toxic effect; Toxicology; Translations; Tretinoin; Validation; Vision; base editing; base editor; biological systems; biomaterial compatibility; cGMP production; cell type; clinical translation; functional restoration; gene correction; gene therapy; genome editing; genotoxicity; in vivo; induced pluripotent stem cell; inward rectifier potassium channel; lead candidate; manufacture; manufacturing facility; manufacturing scale-up; meetings; mouse model; mutation correction; nanocapsule; nonhuman primate; novel therapeutics; pre-Investigational New Drug meeting; pre-clinical; preclinical study; process optimization; programs; rare condition; safety study; scale up; small molecule; subretinal injection; success; therapeutic genome editing

PROJECT SUMMARY/ABSTRACT - LEAD TRAILBLAZER PROJECT 1 Leber Congenital Amaurosis (LCA) is a rare but severe form of pediatric blindness. One subtype, LCA16, is caused by several single-point mutations in the KCNJ13 gene, which encodes the inwardly-rectifying potassium channel Kir7.1 in the retina. There is no FDA-approved treatment for ultra-rare conditions such as LCA16. The objective of Lead Project 1 is to develop a new LCA16 gene therapy utilizing a CRISPR base editor (BE) delivered to the retinal pigment epithelium (RPE) via a nonviral silica nanocapsule (SNC). The SNCs possess many desirable properties, including high delivery efficiency, versatile surface chemistry for ligand conjugation, small particle sizes, good biocompatibility, and scalable production. In preliminary studies, we show that the SNC can transiently deliver a wide range of biologics, including a BE to RPE cells in mice and LCA16 patient-derived induced pluripotent stem cell (iPSC)-RPE. Within five years, we seek to optimize and validate a lead candidate, SNC-101, progress to scaled-up and CGMP production of the product to enable non-human primate (NHP) studies and file an Investigational New Drug (IND) application to the FDA for base editing therapy of LCA16. In Aim 1, we will generate a preclinical validation package with a lead SNC formulation for W53* KCNJ13 correction. We will first optimize the amount of ATRA targeting ligand (RPE cell-specific) and the modified KCNJ13 sgRNA in vitro via the Human Cell Assays Core and in vivo in a W53* LCA16 mouse model. We will then develop a scale-up production process for the optimized SNC (i.e., SNC101) for preclinical studies. We plan to complete one INTERACT meeting at the end of this aim. In Aim 2, we will determine the gene correction efficiency, dosing, and toxicity in mouse models and NHPs. We will comprehensively evaluate the immune, structural, and functional consequences of subretinal delivery of SNC-101 through our W53* LCA16 mouse model. Using our Large Animal Core, we will also perform dose-escalation safety studies in NHPs. Alongside the species comparison, the study will ascertain storage stability, dosing, and toxicity profiles in detail, an important milestone to support an IND with the FDA. In Aim 3, we will conduct IND enabling studies of SNC-101 for W53* KCNJ13 correction. A pre-IND meeting package will be submitted to FDA CBER with the help of our Regulatory Core. Waisman Biomanufacturing will develop a full-scale engineering run to be used in GLP toxicology study in rats and NHPs. Concurrent with the toxicology study, one entire CGMP batch will be manufactured. We will submit an IND application to the FDA in the final year. To date, no nonviral genome editing therapeutic has reached an IND in the eye. Success here with SNC-101 would pave the way for this new therapeutic modality. Finally, the knowledge gained along this development path will accelerate the translation of other nonviral genome editing leads, including SNC-201 and TAGE-101, in the Follower Projects of the CRISPR Vision Program.

项目总结/摘要-领先的Trailblazer项目1 Leber先天性黑蒙（LCA）是一种罕见但严重的儿童失明形式。其中一种亚型LCA 16， 由KCNJ 13基因中的几个单点突变引起，该基因编码内向整流钾 视网膜Kir7.1通道。目前还没有FDA批准的治疗超罕见疾病的方法，如LCA 16。的 牵头项目1的目标是利用CRISPR碱基编辑器（BE）开发新的LCA 16基因疗法。 通过非病毒二氧化硅纳米胶囊（SNC）递送至视网膜色素上皮（RPE）。SNC拥有 许多所需的性质，包括高递送效率，用于配体缀合的通用表面化学， 粒径小，生物相容性好，可规模化生产。在初步研究中，我们表明，SNC 可以瞬时递送广泛的生物制剂，包括BE至小鼠中的RPE细胞和LCA 16患者来源的 诱导多能干细胞（iPSC）-RPE。在五年内，我们寻求优化和验证领先的候选人， SNC-101，该产品的规模扩大和CGMP生产进展，以使非人灵长类动物（NHP） 研究并向FDA提交研究性新药（IND）申请，用于LCA 16的基础编辑治疗。在 目标1，我们将生成一个临床前验证包，其中包含用于W53* KCNJ 13校正的主要SNC制剂。 我们将首先优化ATRA靶向配体（RPE细胞特异性）和修饰的KCNJ 13 sgRNA的量， 通过人细胞测定核心进行体外试验，并在W53* LCA 16小鼠模型中进行体内试验。然后我们将开发一个 优化SNC的按比例放大生产工艺（即，SNC 101）进行临床前研究。我们计划完成 一个互动会议结束时，这一目标。在目标2中，我们将确定基因校正效率、剂量， 以及小鼠模型和NHP的毒性。我们将全面评估免疫，结构， 通过我们的W53* LCA 16小鼠模型视网膜下递送SNC-101的功能后果。使用我们 大型动物中心，我们还将在NHP中进行剂量递增安全性研究。除了物种 相比之下，该研究将详细确定储存稳定性，剂量和毒性特征，这是一个重要的里程碑 支持FDA的IND申请在目标3中，我们将进行SNC-101用于W53* KCNJ 13的IND启动研究 纠正一下在我们的监管核心的帮助下，将向FDA CBER提交IND前会议包。 Waisman Biomanufacturing将开发一个全规模工程运行，用于大鼠GLP毒理学研究 NHPs。在毒理学研究的同时，将生产一个完整的CGMP批次。我们将提交 在最后一年向FDA提交IND申请。到目前为止，还没有非病毒基因组编辑治疗达到 IND在眼睛里。SNC-101的成功将为这种新的治疗方式铺平道路。最后 沿着这条发展道路获得的知识将加速其他非病毒基因组编辑的翻译 包括SNC-201和TAGE-101在内的CRISPR愿景项目的后续项目。