Postnatal and Prenatal Therapeutic Base Editing for Metabolic Diseases

项目总结 开发新的治疗方式的潜力已经给基因组编辑领域注入了活力，因为 它最早出现在20世纪90年代，特别是在演示了可编程基因组编辑之后 2012年由多个团体发布的CRISPR-CAS9。体外治疗已经取得了实质性进展。 基因组编辑在过去几年中在患者中的应用，最引人注目的是使用CAR-T免疫疗法治疗 癌症和对血红蛋白疾病的持久治疗。体内治疗应用的进展，即， 由于基因组传递过程中固有的技术挑战，体细胞基因组编辑速度一直较慢。 将工具编辑到身体中。截至撰写本文时，已发表的成功基因组的例子还很少。 编辑在灵长类动物(包括人类)体内进行，几乎所有的例子都涉及体细胞基因组 肝脏编辑：由脂质纳米粒(LNPs)、PCSK9和Angptl3递送的带有Cas9核酸酶的TTR 腺嘌呤碱基编辑由LNPs提供，而PCSK9与巨核酸酶由腺相关蛋白提供 病毒(AAV)载体。基因组编辑疗法的前景延伸到出生前和子宫内基因组 编辑有可能治疗导致重大发病率和死亡率的遗传性疾病 出生后不久。尽管到目前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、肺、心脏和其他器官中。 我们的整个计划寻求在这些早期成功的基础上，追求将产生重大影响的目标 在推进治疗性基因组编辑领域。我们的三个研究项目寻求发展基地编辑 针对肝脏的治疗，以治疗三种罕见的代谢性遗传病：苯丙酮尿症(PKU)， 遗传性酪氨酸血症1型(HT1)和粘多糖病1型(MPSI)。Lead Project 1将专注于 基于LNP的北京大学产后治疗，目标是在五年结束前提交IND申请 在资助期内，并在不久之后开始1/2期临床试验。项目2将侧重于基于LNP的产后 HT1的治疗，目的是提交IND申请并开始临床试验，以及产前治疗 HT1，目的是在五年资助期内进行临床前研究，以实现最终的IND 如果出生后临床试验被证明是成功的，则申请。项目3将侧重于以AAV为基础的产后和 MPSI的产前治疗，与项目2的目标相似。专注于独特的、专门的资源核心 为了实现这些目标，对小动物和大动物进行非目标编辑和宫内治疗将是不可或缺的。

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.