Alfares Ahmad, Alfadhel Majid, Mujamammi Ahmed, Alotaibi Batoul, Albahkali Sarah, Al Balwi Mohammed, Benabdelkamel Hicham, Masood Afshan, Ali Rizwan, Almuaysib Amani, Al Mahri Saeed, Mohammad Sameer, Alanazi Ibrahim O, Alfadda Assim, AlGhamdi Saleh, Alrfaei Bahauddeen M
Department of Pediatrics, College of Medicine, Qassim University, Al-Qassim, Saudi Arabia.
Department of Pathology and Laboratory Medicine, King Abdulaziz Medical City, MNGHA, Riyadh, Saudi Arabia.
Front Cell Dev Biol. 2020 Jan 10;7:365. doi: 10.3389/fcell.2019.00365. eCollection 2019.
Very-long-chain acyl-coenzyme A dehydrogenase (VLCAD) is a coenzyme encoded by that converts very-long-chain fatty acids into energy. This process is disrupted by c.65C > A; p.Ser22 mutation. To clarify mechanisms by which this mutation leads to VLCAD deficiency, we evaluated differences in molecular and cellular functions between mesenchymal stem cells with normal and mutant VLCAD. Saudi Arabia have a high incidence of this form of mutation. Stem cells with mutant VLCAD were isolated from skin of two patients. Metabolic activity and proliferation were evaluated. The Same evaluation was repeated on normal stem cells introduced with same mutation by CRISPR. Mitochondrial depiction was done by electron microscope and proteomic analysis was done on patients' cells. Metabolic activity and proliferation were significantly lower in patients' cells. Introducing the same mutation into normal stem cells resulted in the same defects. We detected mitochondrial abnormalities by electron microscopy in addition to poor wound healing and migration processes in mutant cells. Furthermore, in a proteomic analysis, we identified several upregulated or downregulated proteins related to hypoglycemia, liver disorder, and cardiac and muscle involvement. We concluded experimental assays of mutant (c.65C > A; p.Ser22) contribute to severe neonatal disorders with hypoglycemia, liver disorder, and cardiac and muscle involvement.
极长链酰基辅酶A脱氢酶（VLCAD）是一种由[具体基因]编码的辅酶，可将极长链脂肪酸转化为能量。该过程因c.65C>A；p.Ser22突变而受到干扰。为阐明这种突变导致VLCAD缺乏的机制，我们评估了具有正常和突变VLCAD的间充质干细胞在分子和细胞功能上的差异。沙特阿拉伯这种形式的突变发生率很高。从两名患者的皮肤中分离出具有突变VLCAD的干细胞。评估了代谢活性和增殖情况。对通过CRISPR引入相同突变的正常干细胞重复进行相同评估。通过电子显微镜进行线粒体描绘，并对患者细胞进行蛋白质组分析。患者细胞的代谢活性和增殖明显较低。将相同突变引入正常干细胞会导致相同缺陷。除了突变细胞中伤口愈合和迁移过程不佳外，我们通过电子显微镜检测到线粒体异常。此外，在蛋白质组分析中，我们鉴定出了几种与低血糖、肝脏疾病以及心脏和肌肉受累相关的上调或下调蛋白质。我们得出结论，对突变体（c.65C>A；p.Ser22）的实验分析导致了伴有低血糖、肝脏疾病以及心脏和肌肉受累的严重新生儿疾病。
