Human genetic variation in PPARGC1B has been associated with adiposity, but the genetic variants that affect PPARGC1B expression have not been experimentally determined. Here, guided by previous observational data, we used CRISPR/Cas9 to scarlessly edit the alleles of the candidate causal genetic variant rs10071329 in a human brown adipocyte cell line (hBAs). Switching the rs10071329 genotype from A/A to G/G enhanced PPARGC1B expression throughout the adipogenic differentiation, identifying rs10071329 as a cis-eQTL. The higher PPARGC1B expression in G/G cells coincided with greater accumulation of triglycerides, and higher expression of mitochondria-encoded genes, but without significant effects on adipogenic marker expression. Furthermore, G/G cells had improved basal- and norepinephrine-stimulated mitochondrial respiration, possibly relating to enhanced mitochondrial gene expression. The G/G cells also exhibited increased norepinephrine-stimulated glycerol release, indicating improved lipolysis. Altogether, our results showed that rs10071329 is a cis-eQTL, with the G/G genotype conferring enhanced PPARGC1B expression, with consequent improved mitochondrial function and response to norepinephrine in brown adipocytes. This genetic variant, and as yet undetermined eQTLs, at PPARGC1B could prove useful in genotype-based precision medicine for obesity treatment.

中文翻译：

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rs10071329 的人类遗传变异与肥胖相关性状相关，调节 PPARGC1B 表达并改变棕色脂肪细胞功能

PPARGC1B 的人类遗传变异与肥胖有关，但影响 PPARGC1B 表达的遗传变异尚未通过实验确定。在此，在先前观察数据的指导下，我们使用 CRISPR/Cas9 无痕编辑人类棕色脂肪细胞系 (hBA) 中候选因果遗传变异 rs10071329 的等位基因。将 rs10071329 基因型从 A/A 转换为 G/G 增强了整个脂肪形成分化过程中 PPARGC1B 的表达，将 rs10071329 鉴定为顺式 eQTL。G/G 细胞中较高的 PPARGC1B 表达与甘油三酯的较高积累和线粒体编码基因的较高表达相一致，但对脂肪形成标志物表达没有显着影响。此外，G/G 细胞改善了基础和去甲肾上腺素刺激的线粒体呼吸，这可能与增强的线粒体基因表达有关。G/G 细胞还表现出去甲肾上腺素刺激的甘油释放增加，表明脂肪分解得到改善。总而言之，我们的结果表明 rs10071329 是一个顺式 eQTL，其 G/G 基因型赋予 PPARGC1B 表达增强，从而改善棕色脂肪细胞中的线粒体功能和对去甲肾上腺素的反应。PPARGC1B 的这种遗传变异以及尚未确定的 eQTL 可能在基于基因型的肥胖治疗精准医学中发挥作用。