Jiang Yanhong, Chen Shuanghong, Hsiao Shenlin, Zhang Haokun, Xie Da, Wang Zi Jun, Ren Wendan, Liu Mingyao, Liao Jiaoyang, Wu Yuxuan
Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai 200241, China; Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai 200241, China.
YolTech Therapeutics, Shanghai 201109, China.
Mol Ther. 2025 Jan 8;33(1):104-118. doi: 10.1016/j.ymthe.2024.10.003. Epub 2024 Oct 9.
Primary hyperoxaluria type 1 (PH1) is a severe genetic metabolic disorder caused by mutations in the AGXT gene, leading to defects in enzymes crucial for glyoxylate metabolism. PH1 is characterized by severe, potentially life-threatening manifestations due to excessive oxalate accumulation, which leads to calcium oxalate crystal deposits in the kidneys and, ultimately, renal failure and systemic oxalosis. Existing substrate reduction therapies, such as inhibition of liver-specific glycolate oxidase (GO) encoded by HAO1 using siRNA or CRISPR-Cas9 delivered by adeno-associated virus, either require repeated dosing or have raised safety concerns. To address these limitations, our study employed lipid nanoparticles (LNPs) for CRISPR-Cas9 delivery to rapidly generate a PH1 mouse model and validate the therapeutic efficacy of LNP-CRISPR-Cas9 targeting the Hao1 gene. The LNP-CRISPR-Cas9 system exhibited efficient editing of the Hao1 gene, significantly reducing GO expression and lowering urinary oxalate levels in treated PH1 mice. Notably, these effects persisted for 12 months with no significant off-target effects, liver-induced toxicity, or substantial immune responses, highlighting the approach's safety and specificity. Furthermore, the developed humanized mouse model validated the efficacy of our therapeutic strategy. These findings support LNP-CRISPR-Cas9 targeting HAO1 as a promising and safer alternative for PH1 treatment with a single administration.
1型原发性高草酸尿症（PH1）是一种严重的遗传性代谢紊乱疾病，由AGXT基因突变引起，导致乙醛酸代谢关键酶缺陷。PH1的特征是由于草酸盐过度积累而出现严重的、可能危及生命的表现，这会导致草酸钙晶体在肾脏中沉积，最终导致肾衰竭和全身性草酸osis。现有的底物还原疗法，如使用小干扰RNA（siRNA）或腺相关病毒递送的CRISPR-Cas9抑制HAO1编码的肝脏特异性乙醇酸氧化酶（GO），要么需要重复给药，要么引发了安全担忧。为了解决这些局限性，我们的研究采用脂质纳米颗粒（LNP）递送CRISPR-Cas9，以快速生成PH1小鼠模型，并验证靶向Hao1基因的LNP-CRISPR-Cas9的治疗效果。LNP-CRISPR-Cas9系统对Hao1基因表现出高效编辑，显著降低了治疗的PH1小鼠的GO表达并降低了尿草酸水平。值得注意的是，这些效果持续了12个月，没有明显的脱靶效应、肝脏诱导的毒性或实质性免疫反应，突出了该方法的安全性和特异性。此外，开发的人源化小鼠模型验证了我们治疗策略的疗效。这些发现支持将靶向HAO1的LNP-CRISPR-Cas9作为一种有前景且更安全的单次给药治疗PH1的替代方法。
