Postnatal and Prenatal Therapeutic Base Editing for Metabolic Diseases

项目概要 自那时起，开发新型治疗方式的潜力就为基因组编辑领域注入了活力。 它首次出现于 20 世纪 90 年代，特别是自从可编程基因组编辑的演示之后 2012年多个研究小组研究CRISPR-Cas9，离体治疗取得实质性进展 过去几年基因组编辑在患者中的应用，最引人注目的是 CAR-T 免疫疗法 癌症和血红蛋白病的持久治疗。体内治疗应用的进展，即 由于基因组传递固有的技术挑战，体细胞基因组编辑的速度较慢 编辑工具进入正文。截至撰写本文时，已发表的成功基因组实例还很少 在灵长类动物（包括人类）体内进行编辑，几乎所有例子都涉及体细胞基因组 肝脏中的编辑：使用脂质纳米粒子 (LNP)、PCSK9 和 ANGPTL3 传递的 Cas9 核酸酶进行 TTR 具有由 LNP 提供的腺嘌呤碱基编辑器，以及具有由腺相关提供的大范围核酸酶的 PCSK9 病毒（AAV）载体。基因组编辑疗法的前景延伸至出生前，子宫内基因组 编辑有可能治疗在之前或之前导致显着发病率和死亡率的遗传疾病 出生后不久。尽管迄今为止仅限于小动物模型，但子宫内基因组编辑已被证明是有效的 在肝脏、肺、心脏和其他器官中。 我们的总体计划旨在以这些早期成功为基础，追求具有重大影响的目标 推进治疗性基因组编辑领域。我们的三个研究项目旨在开发碱基编辑 针对肝脏的疗法可治疗三种罕见的代谢遗传病：苯丙酮尿症 (PKU)、 遗传性酪氨酸血症 1 型 (HT1) 和粘多糖贮积症 1 型 (MPSI)。主导项目 1 将重点关注 基于LNP的PKU产后治疗，目标是5年末提交IND申请 资助期并随后开始1/2期临床试验。项目2将重点关注基于LNP的产后 HT1的治疗，旨在提交IND申请并开始临床试验，以及产前治疗 HT1，旨在在五年资助期内进行临床前研究，以实现最终的 IND 如果产后临床试验成功则可以申请。项目 3 将重点关注基于 AAV 的产后和 MPSI 的产前治疗，与项目 2 具有相似的目标。独特、专业的资源核心侧重于 为了实现这些目标，对小型和大型动物的脱靶编辑和子宫内治疗将是必不可少的。

PROJECT SUMMARY The potential for the development of novel therapeutic modalities has energized the genome editing field since it first emerged in the 1990s and especially since the demonstration of programmable genome editing with CRISPR-Cas9 by multiple groups in 2012. There has been substantial progress with ex vivo therapeutic applications of genome editing in patients in the past few years, most notably with CAR-T immunotherapies for cancer and with durable treatment of hemoglobinopathies. Progress with in vivo therapeutic applications, i.e., somatic cell genome editing, has been slower due to the technical challenges inherent in the delivery of genome- editing tools into the body. As of the time of this writing, there are few published examples of successful genome editing performed in vivo in primates (including humans), with almost all examples involving somatic genome editing in the liver: TTR with Cas9 nuclease delivered by lipid nanoparticles (LNPs), PCSK9 and ANGPTL3 with adenine base editors delivered by LNPs, and PCSK9 with meganucleases delivered by adeno-associated virus (AAV) vectors. The prospects for genome-editing therapies extend to before birth, with in utero genome editing having the potential to treat genetic diseases that result in significant morbidity and mortality before or shortly after birth. Although restricted to small animal models so far, in utero genome editing has proven effective in the liver, lungs, heart, and other organs. Our Overall Program seeks to build on these early successes, pursuing goals that that would be of major impact in advancing the field of therapeutic genome editing. Our three Research Projects seek to develop base-editing therapies targeting the liver in order to treat three rare metabolic genetic diseases: phenylketonuria (PKU), hereditary tyrosinemia type 1 (HT1), and mucopolysaccharidosis type 1 (MPSI). Lead Project 1 will focus on LNP-based postnatal treatment of PKU, with the aim to file an IND application by the end of the five-year funding period and begin a phase 1/2 clinical trial soon afterwards. Project 2 will focus on LNP-based postnatal treatment of HT1, with the aim to file an IND application and begin a clinical trial, and prenatal treatment of HT1, with the aim of performing preclinical studies during the five-year funding period to enable an eventual IND application if the postnatal clinical trial proves successful. Project 3 will focus on AAV-based postnatal and prenatal treatment of MPSI, with similar aims as Project 2. Unique, specialized Resource Cores focused on off-target editing and in utero treatment of small and large animals will be indispensable in achieving these aims.