Erythroid-Specific Expression of LIN28A Is Sufficient for Robust Gamma-Globin Gene and Protein Expression in Adult Erythroblasts.
Erythroid-Specific Expression of LIN28A Is Sufficient for Robust Gamma-Globin Gene and Protein Expression in Adult Erythroblasts.
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DOI：
10.1371/journal.pone.0144977
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Miller JL
中科院分区：
文献类型：
--
作者：
Lee YT;de Vasconcellos JF;Byrnes C;Kaushal M;Rabel A;Tumburu L;Allwardt JM;Miller JL
提高成年人胎儿血红蛋白 (HbF) 水平仍然是血液学研究的一个活跃领域。在这里，我们探讨了红细胞特异性 LIN28A 表达在调节培养的成红细胞中的 γ-珠蛋白基因表达和 HbF 水平方面的作用。为此，生产了慢病毒转导载体，其中 LIN28A 表达由人 KLF1 或 SPTA1 基因的红细胞特异性基因启动子区域驱动。具有连锁的嘌呤霉素抗性标记的 LIN28A 的转基因表达仅限于红系谱系，如红系集落的选择性存活（大于所有集落的 95%）所证明的。成红细胞 LIN28A 过表达 (LIN28A-OE) 不会显着影响增殖或抑制分化。在 LIN28A-OE 细胞中证实总 let-7 microRNA 水平被抑制超过 70%。 γ-珠蛋白 mRNA 和蛋白质表达增加，HbF 水平达到 30-40%。这些数据表明，针对 LIN28A 表达的成红细胞足以增加成人成红细胞中的胎儿血红蛋白表达。
Increasing fetal hemoglobin (HbF) levels in adult humans remains an active area in hematologic research. Here we explored erythroid-specific LIN28A expression for its effect in regulating gamma-globin gene expression and HbF levels in cultured adult erythroblasts. For this purpose, lentiviral transduction vectors were produced with LIN28A expression driven by erythroid-specific gene promoter regions of the human KLF1 or SPTA1 genes. Transgene expression of LIN28A with a linked puromycin resistance marker was restricted to the erythroid lineage as demonstrated by selective survival of erythroid colonies (greater than 95% of all colonies). Erythroblast LIN28A over-expression (LIN28A-OE) did not significantly affect proliferation or inhibit differentiation. Greater than 70% suppression of total let-7 microRNA levels was confirmed in LIN28A-OE cells. Increases in gamma-globin mRNA and protein expression with HbF levels reaching 30–40% were achieved. These data suggest that erythroblast targeting of LIN28A expression is sufficient for increasing fetal hemoglobin expression in adult human erythroblasts.