Department of Pediatric Urology, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Children's Stone Treatment Center of National Health and Family Planning Commission of the People's Republic of China, Shanghai, People's Republic of China.
Am J Physiol Renal Physiol. 2021 Mar 1;320(3):F475-F484. doi: 10.1152/ajprenal.00514.2020. Epub 2021 Jan 25.
Primary hyperoxaluria type 1 (PH1) is a severe inherited disorder caused by a genetic defect in alanine-glyoxylate aminotransferase (), which results in recurrent urolithiasis and renal failure. Animal models that precisely reflect human PH1 phenotypes are lacking. We aimed to develop a novel PH1 rat model and study the mechanisms involved in PH1 deterioration. One cell stage Sprague-Dawley embryos were injected with the CRISPR/Cas9 system to introduce a Q84X mutation in . Liver tissues were harvested to determine expression. Urine oxalate, crystals, and electrolyte levels in and wild-type (WT) littermates were evaluated. Kidney tissues were used for Pizzolato staining and kidney injury evaluation. Data showed that mRNA and protein were absent in rats. At 4 and 24 wk, rats displayed 2.1- and 2.9-fold higher urinary oxalate levels, respectively, compared with WT littermates. As a result, calcium oxalate (CaOx) crystals in urine were revealed in all rats but in none of the WT rats. We also observed bladder stones in 36.4% of rats, of which 44.4% had renal CaOx deposition. Moreover, the elevated serum urea and creatinine levels indicated the impaired renal function in rats. Further investigation revealed significantly increased expression of inflammation-, necroptosis-, and fibrosis-related genes in the kidneys of rats with spontaneous renal CaOx deposition, indicating that these pathways are involved in PH1 deterioration. Collectively, these results suggest that this rat model has broad applicability in mechanistic studies and innovative therapeutics development for PH1 and other kidney stone diseases. Primary hyperoxaluria type 1 is a severe inherited disorder that results in recurrent urolithiasis and renal failure. We generated an alanine-glyoxylate aminotransferase () nonsense mutant rat model that displayed an early onset of hyperoxaluria, spontaneous renal CaOx precipitation, bladder stone, and kidney injuries. Our results suggest an interaction of renal CaOx crystals with the activation of inflammation-, fibrosis-, and necroptosis-related pathways. In all, the rat strain has broad applicability in mechanistic studies and the development of innovative therapeutics.
原发性高草酸尿症 1 型（PH1）是一种由丙氨酸-乙醛酸氨基转移酶（）遗传缺陷引起的严重遗传性疾病，导致反复尿路结石和肾衰竭。缺乏准确反映人类 PH1 表型的动物模型。我们旨在开发一种新型 PH1 大鼠模型，并研究 PH1 恶化涉及的机制。将 Sprague-Dawley 胚胎的单细胞期注射 CRISPR/Cas9 系统，以在中引入 Q84X 突变。收集肝组织以确定的表达。评估和野生型（WT）同窝仔鼠的尿草酸盐、晶体和电解质水平。使用肾组织进行 Pizzolato 染色和肾损伤评估。数据显示，在 PH1 大鼠中不存在的 mRNA 和蛋白。在 4 和 24 周时，与 WT 同窝仔鼠相比，PH1 大鼠的尿草酸盐水平分别高出 2.1 和 2.9 倍。结果，所有 PH1 大鼠的尿液中均显示出钙草酸钙（CaOx）晶体，但在 WT 大鼠中均未显示。我们还观察到 36.4%的 PH1 大鼠存在膀胱结石，其中 44.4%的大鼠有肾 CaOx 沉积。此外，血清尿素和肌酐水平升高表明 PH1 大鼠的肾功能受损。进一步研究表明，在自发肾 CaOx 沉积的 PH1 大鼠肾脏中，炎症、坏死性凋亡和纤维化相关基因的表达显著增加，表明这些途径参与了 PH1 的恶化。总之，这些结果表明，该大鼠模型在 PH1 和其他肾结石疾病的机制研究和创新治疗药物开发中具有广泛的适用性。原发性高草酸尿症 1 型是一种严重的遗传性疾病，导致反复尿路结石和肾衰竭。我们生成了一种丙氨酸-乙醛酸氨基转移酶（）无义突变大鼠模型，该模型表现出早发性高草酸尿症、自发性肾 CaOx 沉淀、膀胱结石和肾脏损伤。我们的结果表明，肾 CaOx 晶体与炎症、纤维化和坏死性凋亡相关途径的激活相互作用。总之，该 PH1 大鼠品系在机制研究和创新治疗药物的开发中有广泛的适用性。
