Quantitative morphology of induced phenotypes in cultured cells and tissues

培养细胞和组织中诱导表型的定量形态学

• 批准号: 8336691
• 负责人: Ilya Goldberg
• 金额: $ 35.16万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8336691
• 关键词: Age; Aging; Aging-Related Process; Animal Model; Appearance; Biological Assay; Biological Markers; Biological Process; Biology of Aging; Caenorhabditis elegans; Candidate Disease Gene; Cell Line; Cell Lineage; Cells; Cellular Morphology; Collaborations; Collection; Computer software; Controlled Environment; Development; Disease; ES Cell Line; Gene Family; Genes; Genetic; Goals; Grouping; Growth; Human; Image; Individual; Libraries; Longevity; Methodology; Methods; Microarray Analysis; Modeling; Molecular; Molecular Probes; Molecular Profiling; Morphology; Mus; Pathway interactions; Phase; Phenotype; Physiological; Population; Publications; Publishing; RNA Interference; Screening procedure; Sorting - Cell Movement; Staging; Techniques; Time; Tissues; Transcription factor genes; Visual; Work; aging gene; aging population; base; embryonic stem cell; functional group; gene function; high throughput screening; image processing; molecular marker; research study; stem cell differentiation; tissue/cell culture; transcription factor

We have continued our work in collaboration with Dr. Minoru Ko (LG-NIA) to characterize developmental pathways induced in mouse embryonic stem cells using transcription factor manipulation. We were able to establish a differentiation assay based on morphology alone, using phase-contrast imaging without specific markers. We were able to show that differentiation can be tracked over time, and that morphological similarities between differentiating cells are established early and maintained for several days of differentiation. An important milestone is that we were able to establish a conditions where cell lines showed consistent grouping by gene function over several independent experiments. Additionally, we were able to expand this analysis to compare the morphologies of 56 cell lines. These results are being prepared for publication. We have continued work characterizing the molecular basis of morphological age-state transitions during the C. elegans life-span. In previous published work we used WND-CHARM to identify distinct morphological aging states in C. elegans. We used this technique to sort worms based on their age state during a transition period where an aging population is evenly divided between individuals in Stage I and Stage II. The worms were identical genetically, by chronological age, by growth conditions, and by visual appearance, and could only be sorted into age-states using WND-CHARM. Micro-array experiments performed on these two sub-populations revealed several hundred genes with significantly altered expression profiles. By comparing our gene lists with those from other aging studies in C. elegans, we were able to identify several gene families and functional groups that were unique to our study, as well as several important aging genes that were identified previously. The results of this study are being prepared for publication. In the next stage of this study, we will begin to screen through our gene list using RNAi libraries to shift the timing of the Stage I - Stage II transition.

我们继续与 Minoru Ko 博士 (LG-NIA) 合作，研究利用转录因子操作在小鼠胚胎干细胞中诱导的发育途径。 我们能够仅基于形态学建立一种分化测定法，使用没有特定标记的相差成像。 我们能够证明分化可以随着时间的推移进行追踪，并且分化细胞之间的形态学相似性在早期就已建立并维持了几天的分化。 一个重要的里程碑是，我们能够建立一种条件，使细胞系在几次独立实验中按基因功能显示出一致的分组。 此外，我们还能够扩展此分析以比较 56 种细胞系的形态。 这些结果正在准备发表。 我们继续研究线虫生命周期中形态年龄状态转变的分子基础。在之前发表的工作中，我们使用 WND-CHARM 来识别秀丽隐杆线虫不同的形态衰老状态。 我们使用这种技术根据过渡时期的年龄状态对蠕虫进行分类，其中老龄化人口在第一阶段和第二阶段的个体之间平均分配。 这些蠕虫在基因、年龄、生长条件和视觉外观上都是相同的，并且只能使用 WND-CHARM 将其分类为年龄状态。 对这两个亚群进行的微阵列实验揭示了数百个表达谱显着改变的基因。 通过将我们的基因列表与线虫其他衰老研究中的基因列表进行比较，我们能够识别出我们研究中特有的几个基因家族和功能组，以及之前发现的几个重要的衰老基因。 这项研究的结果正在准备发表。 在本研究的下一阶段，我们将开始使用 RNAi 文库筛选基因列表，以改变第一阶段到第二阶段过渡的时间。

项目成果

Progression analysis and stage discovery in continuous physiological processes using image computing.

• DOI: 10.1155/2010/107036
• 发表时间: 2010
• 期刊: EURASIP journal on bioinformatics & systems biology
• 作者: Shamir L;Rahimi S;Orlov N;Ferrucci L;Goldberg IG
• 通讯作者: Goldberg IG

CENP-V is required for centromere organization, chromosome alignment and cytokinesis.

• DOI: 10.1038/emboj.2008.175
• 发表时间: 2008-10-08
• 期刊: EMBO JOURNAL
• 影响因子: 11.4
• 作者: Tadeu, Ana Mafalda Baptista;Ribeiro, Susana;Johnston, Josiah;Goldberg, Ilya;Gerloff, Dietlind;Earnshaw, William C.
• 通讯作者: Earnshaw, William C.

Early detection of radiographic knee osteoarthritis using computer-aided analysis.

• DOI: 10.1016/j.joca.2009.04.010
• 发表时间: 2009-10
• 期刊: Osteoarthritis and cartilage
• 影响因子: 7
• 作者: Shamir L;Ling SM;Scott W;Hochberg M;Ferrucci L;Goldberg IG
• 通讯作者: Goldberg IG