A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts.
A new physiological medium uncovers biochemical and cellular alterations in Lesch-Nyhan disease fibroblasts.
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DOI：
10.1186/s10020-023-00774-8
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发表时间：
2024-01-03
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Lesch-Nyhan病(LND)是由次黄嘌呤鸟嘌呤磷酸核糖基转移酶(HGprt)基因缺陷引起的一种严重的神经系统疾病，HGprt参与嘌呤的补救合成。为了弥补这一缺陷，从头合成嘌呤的生物合成途径被加速。大多数研究都没有发现从患者身上获得的培养细胞中有任何持续的嘌呤核苷酸异常。最近的研究表明，新生途径的中间体5-氨基咪唑-4-甲酰胺核苷-5-ʹ-单磷酸(ZMP)在含有生理水平(25nM)叶酸(FA)的LND成纤维细胞中积累，这与普通细胞培养液中的FA水平(2200nM)有很大的不同。然而，RPMI和其他标准介质含有许多非生理性水平的许多营养物质，对细胞代谢有很大影响，并不能准确概括细胞在体内的行为。我们准备了一种新的含有包括维生素在内的所有营养物质的生理水平的培养基来研究可能已经被非生理性介质使用所掩盖的LND成纤维细胞的潜在变化。我们量化了不同培养条件下ZMP的积累，并评估了两个已知的ZMP靶蛋白(AMPK和ADSL)的活性，叶酸载体SLC19A1的mRNA表达，可能的线粒体变化和LND成纤维细胞的功能后果。与对照组相比，与对照组相比，用Platmax-PV维持的LND成纤维细胞表现出代谢适应，如更高的糖酵解能力，叶酸载体SCL19A1的表达增加，以及功能变化，如线粒体潜力降低和细胞迁移减少。这些变化可以通过高水平的叶酸逆转，这表明叶酸补充剂可能是一种潜在的LND治疗方法。一种完整的生理细胞培养液揭示了Lesch-Nyhan病的新变化。这项工作强调了在研究代谢紊乱时使用生理细胞培养条件的重要性。网上版载有补充材料，可在10.1186/s10020023-00774-8查阅。
Lesch-Nyhan disease (LND) is a severe neurological disorder caused by the genetic deficiency of hypoxanthine–guanine phosphoribosyltransferase (HGprt), an enzyme involved in the salvage synthesis of purines. To compensate this deficiency, there is an acceleration of the de novo purine biosynthetic pathway. Most studies have failed to find any consistent abnormalities of purine nucleotides in cultured cells obtained from the patients. Recently, it has been shown that 5-aminoimidazole-4-carboxamide riboside 5ʹ-monophosphate (ZMP), an intermediate of the de novo pathway, accumulates in LND fibroblasts maintained with RPMI containing physiological levels (25 nM) of folic acid (FA), which strongly differs from FA levels of regular cell culture media (2200 nM). However, RPMI and other standard media contain non-physiological levels of many nutrients, having a great impact in cell metabolism that does not precisely recapitulate the in vivo behavior of cells. We prepared a new culture medium containing physiological levels of all nutrients, including vitamins (Plasmax-PV), to study the potential alterations of LND fibroblasts that may have been masked by the usage of non-physiological media. We quantified ZMP accumulation under different culture conditions and evaluated the activity of two known ZMP-target proteins (AMPK and ADSL), the mRNA expression of the folate carrier SLC19A1, possible mitochondrial alterations and functional consequences in LND fibroblasts. LND fibroblasts maintained with Plasmax-PV show metabolic adaptations such a higher glycolytic capacity, increased expression of the folate carrier SCL19A1, and functional alterations such a decreased mitochondrial potential and reduced cell migration compared to controls. These alterations can be reverted with high levels of folic acid, suggesting that folic acid supplements might be a potential treatment for LND. A complete physiological cell culture medium reveals new alterations in Lesch-Nyhan disease. This work emphasizes the importance of using physiological cell culture conditions when studying a metabolic disorder. The online version contains supplementary material available at 10.1186/s10020-023-00774-8.
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DOI：
10.1073/pnas.2025932118
发表时间：
2021-09-14
影响因子：
11.1
作者：
Drake JC;Wilson RJ;Laker RC;Guan Y;Spaulding HR;Nichenko AS;Shen W;Shang H;Dorn MV;Huang K;Zhang M;Bandara AB;Brisendine MH;Kashatus JA;Sharma PR;Young A;Gautam J;Cao R;Wallrabe H;Chang PA;Wong M;Desjardins EM;Hawley SA;Christ GJ;Kashatus DF;Miller CL;Wolf MJ;Periasamy A;Steinberg GR;Hardie DG;Yan Z
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Yan Z
影响因子：
5.9
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Bell S;McCarty V;Peng H;Jefri M;Hettige N;Antonyan L;Crapper L;O'Leary LA;Zhang X;Zhang Y;Wu H;Sutcliffe D;Kolobova I;Rosenberger TA;Moquin L;Gratton A;Popic J;Gantois I;Stumpf PS;Schuppert AA;Mechawar N;Sonenberg N;Tremblay ML;Jinnah HA;Ernst C
通讯作者：
Ernst C
影响因子：
4.1
作者：
Guigas, Bruno;Taleux, Nellie;Hue, Louis
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Hue, Louis
DOI：
10.3390/antiox11010097
发表时间：
2021-12-30
期刊：
Antioxidants (Basel, Switzerland)
影响因子：
--
作者：
Golikov MV;Karpenko IL;Lipatova AV;Ivanova ON;Fedyakina IT;Larichev VF;Zakirova NF;Leonova OG;Popenko VI;Bartosch B;Kochetkov SN;Smirnova OA;Ivanov AV
通讯作者：
Ivanov AV
DOI：
10.1038/s41577-021-00524-z
发表时间：
2021-09
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
Decout A;Katz JD;Venkatraman S;Ablasser A
通讯作者：
Ablasser A