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.

中文翻译：

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通过 LNP-CRISPR-Cas9 介导的乙醇酸氧化酶破坏对 1 型原发性高草酸尿症进行高效、安全的体内治疗


原发性高草酸尿症 1 型 （PH1） 是一种由 AGXT 基因突变引起的严重遗传代谢疾病，导致对乙醛酸代谢至关重要的酶出现缺陷。PH1 的特征是由于草酸盐积累过多而出现严重的、可能危及生命的表现，这会导致草酸钙晶体沉积在肾脏中，最终导致肾功能衰竭和系统性草酸中毒。现有的底物减少疗法，例如使用 siRNA 或腺相关病毒递送的 CRISPR-Cas9 抑制由 HAO1 编码的肝脏特异性乙醇酸氧化酶 （GO），要么需要重复给药，要么引起安全问题。为了解决这些限制，我们的研究采用脂质纳米颗粒 （LNP） 进行 CRISPR-Cas9 递送，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 系统表现出对 Hao1 基因的有效编辑，显着降低 GO 表达并降低治疗 PH1 小鼠的尿草酸盐水平。值得注意的是，这些影响持续了 12 个月，没有明显的脱靶效应、肝脏诱导的毒性或实质性的免疫反应，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。这些发现支持靶向 HAO1 的 LNP-CRISPR-Cas9 是一种有前途且更安全的 PH1 治疗替代方案，只需一次给药即可。