Elucidation of the Role of FAM210B in Mitochondrial Metabolism and Erythropoiesis
Elucidation of the Role of FAM210B in Mitochondrial Metabolism and Erythropoiesis
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DOI：
10.1128/mcb.00143-22
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发表时间：
2022-11
影响因子：
5.3
通讯作者：
Chie Suzuki;T. Fujiwara;Hiroki Shima;K. Ono;Kei Saito;Hiroki Kato;Koichi Onodera;S. Ichikawa;N. Fukuhara;Y. Onishi;Hisayuki Yokoyama;Y. Nakamura;K. Igarashi;H. Harigae
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文献类型：
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作者：
Chie Suzuki;T. Fujiwara;Hiroki Shima;K. Ono;Kei Saito;Hiroki Kato;Koichi Onodera;S. Ichikawa;N. Fukuhara;Y. Onishi;Hisayuki Yokoyama;Y. Nakamura;K. Igarashi;H. Harigae
线粒体在红系分化过程中起着重要而特异的作用。最近，编码线粒体内膜蛋白的FAM 210 B已被鉴定为加塔-1的新靶点，以及促红细胞生成素诱导基因。线粒体在红系分化过程中起着重要而特殊的作用。最近，编码线粒体内膜蛋白的FAM 210 B已被鉴定为加塔-1的新靶点，以及促红细胞生成素诱导基因。FAM 210 B蛋白参与调节线粒体代谢和血红素生物合成，但其具体功能尚不清楚。在这里，我们使用CRISPR/Cas9方法在人诱导多能干细胞衍生的红系祖细胞（HiDEP）中产生内源性FAM 210 B的敲除和敲低。有趣的是，在FAM 210 B耗尽的细胞中，红系分化更明显，这导致正染性成红细胞的频率增加和嗜碱性/嗜多染性成红细胞的频率降低。综合代谢物分析和功能分析表明，在FAM 210 B缺失的HiDEP细胞中，氧消耗速率和NAD（NAD+）/NADH比率显著降低，而乳酸产生显著增加，表明FAM 210 B参与成红细胞线粒体能量代谢。最后，我们从稳定表达His/生物素标记的FAM 210 B的K562细胞中纯化了FAM 210 B相互作用蛋白。His/生物素纯化的材料的质谱分析表明与线粒体ATP酶的多个亚基，如亚基α（ATP 5A）和β（ATP 5 B）的相互作用。我们的研究结果表明，FAM 210 B通过调节线粒体能量代谢对红系细胞分化有重要作用。我们的研究结果提供了深入了解造血失调的病理生理学。
Mitochondria play essential and specific roles during erythroid differentiation. Recently, FAM210B, encoding a mitochondrial inner membrane protein, has been identified as a novel target of GATA-1, as well as an erythropoietin-inducible gene. ABSTRACT Mitochondria play essential and specific roles during erythroid differentiation. Recently, FAM210B, encoding a mitochondrial inner membrane protein, has been identified as a novel target of GATA-1, as well as an erythropoietin-inducible gene. While FAM210B protein is involved in regulate mitochondrial metabolism and heme biosynthesis, its detailed function remains unknown. Here, we generated both knockout and knockdown of endogenous FAM210B in human induced pluripotent stem-derived erythroid progenitor (HiDEP) cells using CRISPR/Cas9 methodology. Intriguingly, erythroid differentiation was more pronounced in the FAM210B-depleted cells, and this resulted in increased frequency of orthochromatic erythroblasts and decreased frequencies of basophilic/polychromatic erythroblasts. Comprehensive metabolite analysis and functional analysis indicated that oxygen consumption rates and the NAD (NAD+)/NADH ratio were significantly decreased, while lactate production was significantly increased in FAM210B deletion HiDEP cells, indicating involvement of FAM210B in mitochondrial energy metabolism in erythroblasts. Finally, we purified FAM210B-interacting protein from K562 cells that stably expressed His/biotin-tagged FAM210B. Mass spectrometry analysis of the His/biotin-purified material indicated interactions with multiple subunits of mitochondrial ATP synthases, such as subunit alpha (ATP5A) and beta (ATP5B). Our results suggested that FAM210B contributes prominently to erythroid differentiation by regulating mitochondrial energy metabolism. Our results provide insights into the pathophysiology of dysregulated hematopoiesis.