In vivo delivery of Ab-directed CRISPR ribonucleoproteins for anal cancer immunotherapy

项目总结： 今年在美国，预计将有9760人被诊断为肛门癌。食道癌的5年生存率 按标准护理化疗，转移性肛门癌仅占30%左右。FDA还没有批准过任何方案- 化疗失败后批准，单剂免疫仅显示有限的益处 检查站封锁(ICB)。迫切需要新的治疗方案来改善患者的预后。目标是 这个直接到第二阶段的项目是开发Tage-201，一种瘤内注射的抗体(Ab)- 指导CRISPR基因编辑程序敲除TME免疫细胞中的ADAR，以便与ICB结合使用。 这种新的生物是基于Spotlight Treateutics公司专有的靶向活性基因编辑器(TICE) 站台。Tage是一种非病毒、非纳米颗粒的给药方式，由设计的CRISPR-Cas9组成 核糖核蛋白(RNP)与抗体融合，在体内实现细胞靶向传递和基因敲除。阿达尔 (作用于RNA的腺苷脱氨酶)干扰刺激dsRNA传感通路和I型干扰素 反应，这可以驱动肿瘤微环境(TME)的状态转变为免疫许可， 克服ICB产生最佳抗肿瘤免疫反应的主要障碍。Tage启用 ADAR的基因敲除，尽管有令人信服的临床前证据，但很难用药。初步 用优化的抗体和CRISPR-Cas与小鼠替代原型Tage-201的数据表明，Adar 基因敲除联合ICB治疗介导同基因小鼠肿瘤的全身性抗肿瘤反应 不单独对ICB做出响应的型号。此直接到第二阶段的项目旨在1)确定TACGE-201引线 通过优化RNP架构和引导RNA组件候选，2)生成临床前数据包 小鼠肿瘤直接局部注射后TME免疫细胞Adar基因敲除的实验研究 Tage-201的替代物(msTAGE-201)，在多种小鼠和PDX肿瘤模型中的抗肿瘤效果， 符合ADAR生物学的药效学效应，以及药代动力学和毒性研究 展示可接受的安全概况，以及3)与FDA举行互动会议，以介绍 临床前、可制造性评估和初步配方数据包和IND使能 学习计划。在成功完成直通二期项目后，Tage-201开发项目 候选人将准备启动IND使能研究，随后是临床试验，目标是提供 为目前未接受FDA批准的治疗的患者群体提供有效的治疗选择 改善总体结果和生活质量的战略。

PROJECT SUMMARY: This year in the US, 9,760 people are expected to be diagnosed with anal cancer. The 5-year survival rate for metastatic anal cancer is only ~30% with standard of care chemotherapy. No regimens have been FDA- approved after chemotherapy has failed and only limited benefit has been shown with single agent immune checkpoint blockade (ICB). New treatment options are urgently needed to improve patient outcomes. The goal of this Direct to Phase II project is to develop TAGE-201, an intratumorally administered Antibody (Ab)- directed CRISPR gene editor to knockout ADAR in TME immune cells for use in combination with ICB. This new biologic is based on Spotlight Therapeutics’ proprietary Targeted Active Gene Editor (TAGE) platform. TAGE is a non-viral, non-nanoparticle delivery modality, comprised of an engineered CRISPR-Cas9 ribonucleoprotien (RNP) fused to an Ab, enabling cell-targeted delivery and gene knockout in vivo. ADAR (adenosine deaminase acting on RNA) disruption stimulates dsRNA sensing pathways and a type I interferon response, which can drive a shift the state of the tumor microenvironments (TMEs) to be immune permissive, overcoming a major barrier to generating an optimal anti-tumor immune response with ICB. TAGE enables gene knockout of ADAR, which has been difficult to drug despite compelling preclinical evidence. Preliminary data with a murine surrogate prototype of TAGE-201, with optimized Ab and CRISPR-Cas, suggests Adar knockout in conjunction with ICB treatment mediates systemic anti-tumor responses in syngeneic murine tumor models that do not respond to ICB alone. This Direct to Phase II project aims to 1) identify a TAGE-201 lead candidate by optimizing RNP architecture and guide RNA components, 2) generate a preclinical data package demonstrating Adar knockout in TME immune cells of a murine tumor after direct local injection with murine surrogate of TAGE-201 (msTAGE-201), anti-tumor efficacy in multiple murine and PDX tumor models, pharmacodynamics effects that align with ADAR biology, and pharmacokinetics and toxicity studies that demonstrates an acceptable safety profile, and 3) conduct an INTERACT meeting with the FDA to present the preclinical, manufacturability assessment, and preliminary formulation data packages and an IND-enabling study plan. Upon successful completion of the Direct to Phase II project, the TAGE-201 development candidate will be poised to initiate IND-enabling studies, followed by clinical trials, with the goal to provide an effective treatment option for a population of patients that currently do not have FDA-approved therapeutic strategies to improve overall outcomes and quality of life.