Postnatal and Prenatal Therapeutic Base Editing for Metabolic Diseases

项目摘要 自2010年以来，开发新型治疗方式的潜力为基因组编辑领域注入了活力。 它最早出现在20世纪90年代，特别是自从可编程基因组编辑的演示以来， CRISPR-Cas9在2012年被多个小组研究。离体治疗已经取得了实质性进展， 在过去几年中，基因组编辑在患者中的应用，最值得注意的是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， 体细胞基因组编辑，由于基因组递送中固有的技术挑战， 编辑工具到身体。在撰写本文时，很少有成功的基因组 在灵长类动物（包括人类）体内进行的编辑，几乎所有的例子都涉及体细胞基因组 肝脏中的编辑：脂质纳米颗粒（LNP）、PCSK 9和ANGPTL 3递送的Cas9核酸酶的TTR 具有由LNP递送的腺嘌呤碱基编辑器，以及具有由腺相关病毒载体递送的大范围核酸酶的PCSK 9。 病毒（AAV）载体。基因组编辑疗法的前景延伸到出生前，子宫内基因组 编辑有可能治疗导致显著发病率和死亡率的遗传疾病， 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 在肝脏肺心脏和其他器官中 我们的总体计划旨在建立在这些早期成功的基础上，追求将产生重大影响的目标 in advancing推进the field领域of therapeutic治疗genome基因编辑.我们的三个研究项目旨在开发基础编辑 靶向肝脏的疗法，以治疗三种罕见的代谢遗传疾病：苯丙酮尿症（PKU）， 遗传性酪氨酸血症1型（HT 1）和粘多糖样沉积症1型（MPSI）。项目1将侧重于 以LNP为基础的PKU产后治疗，目的是在五年结束前提交IND申请 并在之后不久开始1/2期临床试验。项目2将侧重于基于LNP的产后 治疗HT 1，目的是提交IND申请并开始临床试验，以及产前治疗 HT 1，目的是在五年资助期内进行临床前研究，以实现最终的IND 如果产后临床试验证明成功，则申请。项目3将侧重于基于AAV的产后和 MPSI的产前治疗，目标与项目2相似。独特、专业的资源核心， 脱靶编辑和对小型和大型动物的子宫内治疗对于实现这些目标是必不可少的。

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.