Automated image analysis of nuclear shape: What can we learn from a prematurely aged cell?
Automated image analysis of nuclear shape: What can we learn from a prematurely aged cell?
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DOI：
10.18632/aging.100434
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发表时间：
2012-02-01
期刊：
影响因子：
5.2
通讯作者：
Cao, Kan
中科院分区：
文献类型：
--
作者：
Driscoll, Meghan K.;Albanese, Jason L.;Cao, Kan
过早衰老的疾病，哈钦森-吉尔福德早衰综合征（HGPS），是由突变核纤层蛋白A，影响核支架。HGPS的表型标志是核起泡。有趣的是，在健康个体的衰老细胞中也观察到了类似的核起泡。最近的研究表明，用雷帕霉素（mTOR通路的抑制剂）治疗可减少HGPS成纤维细胞的核起泡。然而，每个细胞群内起泡的程度差异很大，这使得手动盲计数具有挑战性和主观性。在这里，我们展示了一种新型的，自动化的和高通量的核形状分析，定量测量曲率，面积，周长，偏心率和其他指标的核形态的大群体的细胞。我们检测了用雷帕霉素和RAD 001（雷帕霉素的类似物）处理的HGPS成纤维细胞。我们的分析表明，用RAD 001和雷帕霉素处理减少了核起泡，与盲计数对照一致。此外，我们发现，雷帕霉素治疗减少了核的面积，但保持离心率不变。我们的细胞核形状分析为细胞群体提供了一个无偏见的多维“指纹”，可用于量化治疗效果和分析细胞老化。
The premature aging disorder, Hutchinson-Gilford progeria syndrome (HGPS), is caused by mutant lamin A, which affects the nuclear scaffolding. The phenotypic hallmark of HGPS is nuclear blebbing. Interestingly, similar nuclear blebbing has also been observed in aged cells from healthy individuals. Recent work has shown that treatment with rapamycin, an inhibitor of the mTOR pathway, reduced nuclear blebbing in HGPS fibroblasts. However, the extent of blebbing varies considerably within each cell population, which makes manual blind counting challenging and subjective. Here, we show a novel, automated and high throughput nuclear shape analysis that quantitatively measures curvature, area, perimeter, eccentricity and additional metrics of nuclear morphology for large populations of cells. We examined HGPS fibroblast cells treated with rapamycin and RAD001 (an analog to rapamycin). Our analysis shows that treatment with RAD001 and rapamycin reduces nuclear blebbing, consistent with blind counting controls. In addition, we find that rapamycin treatment reduces the area of the nucleus, but leaves the eccentricity unchanged. Our nuclear shape analysis provides an unbiased, multidimensional "fingerprint" for a population of cells, which can be used to quantify treatment efficacy and analyze cellular aging.