摘要
背景: 原发性高草酸尿1型(pH1)是丙氨酸乙醛酸转氨酶(AGXT)突变引起的一种遗传性疾病。其特征是肝脏乙醛酸代谢异常,导致内源性草酸盐过量产生,并在多个器官(主要是肾脏)沉积草酸盐。PH1患者常反复出现尿路结石,最终出现肾功能衰竭。除肝肾联合移植外,没有有效的治疗方法。 方法: 对pH1大鼠进行微量注射。收集尿液样本进行尿液分析。肾T发生分别进行Western blotting、定量PCR、AGT分析和组织学评价。 结果: 在本研究中,我们通过CRISPR/CAS9介导的大鼠线粒体定位agxt基因亚型的破坏,建立了一种新的pH1疾病模型。Agxt缺乏大鼠尿中草酸的排泄量高于野生动物。同时,突变大鼠出现结晶尿,肾小管轻度扩张,肾轻度纤维化。在饮用水中添加0.4%乙二醇(eg)后,突变大鼠排泄出更多的草酸,与野生动物相比出现严重的肾钙化。突变体中也发现炎症和纤维化相关基因的表达显著升高。 结论: 这些数据表明线粒体中的Agxt-缺乏会损害乙醛酸代谢,并导致大鼠PH1。这种大鼠菌株不仅是研究PH1发病机制和病理学的有用模型,而且是开发和评价创新药物和治疗学的重要工具。
关键词: 原发性高草酸尿1型,肾钙化,CRISPR/CAS9,肾脏,基因编辑,线粒体。
Current Molecular Medicine
Title:Generation of a Primary Hyperoxaluria Type 1 Disease Model Via CRISPR/Cas9 System in Rats
Volume: 18 Issue: 7
关键词: 原发性高草酸尿1型,肾钙化,CRISPR/CAS9,肾脏,基因编辑,线粒体。
摘要: Background: Primary hyperoxaluria type 1 (PH1) is an inherited disease caused by mutations in alanine-glyoxylate aminotransferase (AGXT). It is characterized by abnormal metabolism of glyoxylic acid in the liver leading to endogenous oxalate overproduction and deposition of oxalate in multiple organs, mainly the kidney. Patients of PH1 often suffer from recurrent urinary tract stones, and finally renal failure. There is no effective treatment other than combined liver-kidney transplantation.
Methods: Microinjection was administered to PH1 rats. Urine samples were collected for urine analysis. Kidney tissues were for Western blotting, quantitative PCR, AGT assays and histological evaluation.
Results: In this study, we generated a novel PH1 disease model through CRISPR/Cas9 mediated disruption of mitochondrial localized Agxt gene isoform in rats. Agxt-deficient rats excreted more oxalate in the urine than WT animals. Meanwhile, mutant rats exhibited crystalluria and showed a slight dilatation of renal tubules with mild fibrosis in the kidney. When supplied with 0.4% ethylene glycol (EG) in drinking water, mutant rats excreted greater abundance of oxalate and developed severe nephrocalcinosis in contrast to WT animals. Significantly elevated expression of inflammation- and fibrosisrelated genes was also detected in mutants.
Conclusion: These data suggest that Agxt-deficiency in mitochondria impairs glyoxylic acid metabolism and leads to PH1 in rats. This rat strain would not only be a useful model for the study of the pathogenesis and pathology of PH1 but also a valuable tool for the development and evaluation of innovative drugs and therapeutics.
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Cite this article as:
Generation of a Primary Hyperoxaluria Type 1 Disease Model Via CRISPR/Cas9 System in Rats, Current Molecular Medicine 2018; 18 (7) . https://dx.doi.org/10.2174/1566524019666181212092440
DOI https://dx.doi.org/10.2174/1566524019666181212092440 |
Print ISSN 1566-5240 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5666 |
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