通过对细胞形态的关联影响来发现代谢性疾病基因相互作用。
Discovering metabolic disease gene interactions by correlated effects on cellular morphology.
机构信息
Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
University of Cambridge Metabolic Research Laboratories, Institute of Metabolic Science, Addenbrooke's Hospital, Cambridge, CB2 0QQ, UK.
出版信息
Mol Metab. 2019 Jun;24:108-119. doi: 10.1016/j.molmet.2019.03.001. Epub 2019 Mar 13.
OBJECTIVE
Impaired expansion of peripheral fat contributes to the pathogenesis of insulin resistance and Type 2 Diabetes (T2D). We aimed to identify novel disease-gene interactions during adipocyte differentiation.
METHODS
Genes in disease-associated loci for T2D, adiposity and insulin resistance were ranked according to expression in human adipocytes. The top 125 genes were ablated in human pre-adipocytes via CRISPR/CAS9 and the resulting cellular phenotypes quantified during adipocyte differentiation with high-content microscopy and automated image analysis. Morphometric measurements were extracted from all images and used to construct morphologic profiles for each gene.
RESULTS
Over 10 morphometric measurements were obtained. Clustering of the morphologic profiles accross all genes revealed a group of 14 genes characterized by decreased lipid accumulation, and enriched for known lipodystrophy genes. For two lipodystrophy genes, BSCL2 and AGPAT2, sub-clusters with PLIN1 and CEBPA identifed by morphological similarity were validated by independent experiments as novel protein-protein and gene regulatory interactions.
CONCLUSIONS
A morphometric approach in adipocytes can resolve multiple cellular mechanisms for metabolic disease loci; this approach enables mechanistic interrogation of the hundreds of metabolic disease loci whose function still remains unknown.
目的
外周脂肪扩张能力受损是导致胰岛素抵抗和 2 型糖尿病（T2D）发病的原因之一。本研究旨在鉴定脂肪细胞分化过程中的新的疾病基因相互作用。
方法
根据人类脂肪细胞中的表达情况，对与 T2D、肥胖和胰岛素抵抗相关的疾病相关基因座中的基因进行排序。通过 CRISPR/CAS9 技术使人类前脂肪细胞中的前 125 个基因失活，并通过高内涵显微镜和自动图像分析在脂肪细胞分化过程中对所得的细胞表型进行定量。从所有图像中提取形态学测量值，并用于构建每个基因的形态学图谱。
结果
获得了超过 10 项形态学测量值。对所有基因的形态图谱进行聚类分析，发现了一组 14 个基因，其特征是脂质积累减少，并且富含已知的脂肪营养不良基因。对于两个脂肪营养不良基因 BSCL2 和 AGPAT2，通过形态相似性鉴定的 PLIN1 和 CEBPA 亚群通过独立实验得到了验证，这是新的蛋白质-蛋白质和基因调控相互作用。
结论
脂肪细胞中的形态学方法可以解析代谢疾病基因座的多种细胞机制；这种方法能够对数百个代谢疾病基因座的功能进行机制研究，而这些基因座的功能目前仍不清楚。
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