Regulatory Mechanisms Governing Vertebral Segmentation

椎骨分割的调节机制

基本信息

批准号：
9316666
负责人：
Ertugrul M Ozbudak
金额：
$ 35.1万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9316666
关键词：
Address Affect Binding Binding Sites Cell Differentiation process Cell Proliferation Cells Circadian Rhythms Congenital Abnormality Defect Elements Embryonic Development Feedback Gene Expression Gene Family Genes Genetic Genetic Transcription Heterogeneous-Nuclear Ribonucleoproteins Human Informatin Malignant Neoplasms Maps Mass Spectrum Analysis Mathematics Measures Messenger RNA Mission Modeling Mus Mutation Ovary Pacemakers Pathway interactions Pattern Periodicity Process Protein Family Proteins RNA Binding RNA Decay RNA-Binding Proteins Role Segmentation Clock Pathway Stem cells Testing Time Tissue Therapy Tissues Transcription Repressor/Corepressor Ubiquitin United States National Institutes of Health Untranslated Regions Vertebral column Zebrafish base biological systems cancer therapy embryonic stem cell experimental study gain of function gene discovery in vivo loss of function mRNA Instability malformation multicatalytic endopeptidase complex nerve stem cell overexpression predictive modeling prevent protein degradation protein expression public health relevance spine bone structure transcription factor transcriptome sequencing tumor progression

项目摘要

DESCRIPTION (provided by applicant): Gene expression oscillations are widespread in biological systems. The segmentation clock is one such oscillator controlling segmentation of the vertebral column. Its disruption results in congenital vertebral defects in humans. We have built a mathematical pacemaker model that is based on a transcriptional-feedback loop. Our model predicts that a moderate increase in the stabilities of Her proteins or mRNAs should lengthen the period, while a further increase would abolish the oscillations. To establish the mechanisms of Her protein and mRNA turnover, and to elucidate the mechanism that transfers this periodic information to cells of the next- forming segments, we will: 1. Discover the post-translational mechanism that rapidly recycles oscillating proteins: We will perform loss-of- function experiments for proteins that are candidates to regulate rapid degradation of Her proteins. We will assess whether any of these genes regulate the degradation of the Her-family proteins. Finally, to test the prediction of our model, we will stabilize Her proteins to varying levels by reducing expression of proteins that trigger its degradation and measure corresponding changes in the segmentation process. 2. Discover the post-transcriptional mechanism that rapidly recycles oscillating RNAs: We will determine loss- of-function of which her-RNA-binding proteins stabilize her mRNAs and result in vertebral segmentation defects. We will test the prediction of our model by increasing the halflives of her mRNAs through reduced expression of RNA-binding proteins and determining how the segmentation process is affected. 3. Discover the information transfer mechanism from the segmentation clock to the segmentation machinery: We will perform time-resolved overexpression and loss-of-function experiments for the mesp gene to determine its impact on the segmentation process. We will determine the regulatory cascade starting with the segmentation clock, continuing through the dynamically expressed mesp transcription factor and ending with the formation of segment boundaries. Hes proteins also oscillate in neural progenitor cells, ovary cells and embryonic stem cells, where the oscillations appear to control the temporal switch from proliferation to differentiation. Gain-of-function of Hes proteins is correlated with cancer and their inhibition restores differentiation. Elucidating the dynamics of the Hes/Her oscillations during vertebral segmentation is significant not only for understanding and potentially preventing vertebral malformations, but also for developing approaches for controlled stem cell proliferation and differentiation in various tissues and therapies against cancer progression. Therefore, this application has strong relevance to the mission of the National Institute of Health.

描述（由申请人提供）：基因表达振荡在生物系统中广泛。分割时钟是椎骨列的控制分割的振荡器之一。它的破坏会导致人类先天性椎骨缺陷。我们已经建立了基于转录反馈循环的数学起搏器模型。我们的模型预测，她的蛋白质或mRNA的稳定性适度增加应延长时期，而进一步的增加将消除振荡。为了建立其蛋白质和mRNA转换的机制，并阐明将此周期性信息转移到下一个细分细胞的细胞的机制，我们将：1。发现迅速循环循环振荡蛋白质的翻译后机制：我们将对蛋白质造成损失的蛋白质，以调节蛋白质，以调节蛋白质的快速降级。我们将评估这些基因中的任何一个是否调节其家庭蛋白质的降解。最后，为了测试我们的模型的预测，我们将通过减少蛋白质的表达来稳定其蛋白质的水平，从而触发其降解并测量分割过程中的相应变化。 2。发现快速回收振荡RNA的转录后机制：我们将确定其损失的功能，其中Her-RNA结合蛋白可以稳定其mRNA并导致椎骨分割缺陷。我们将通过降低RNA结合蛋白的表达并确定分割过程的影响如何通过增加其mRNA的半镜来测试模型的预测。 3。发现从分割时钟到分割机械的信息传输机制：我们将对MESP基因执行时间分辨的过表达和功能丧失实验，以确定其对分割过程的影响。我们将从分段时钟开始，通过动态表达的MESP转录因子继续进行调节级联，并以段边界的形成结束。 HES蛋白还会在神经祖细胞，卵巢细胞和胚胎干细胞中振荡，其中振荡似乎控制了从增殖到分化的时间切换。 HES蛋白的功能收益与癌症及其抑制作用恢复分化相关。阐明在椎骨分割过程中HES/她的振荡动力学不仅对于理解和潜在预防椎骨畸形，而且对于开发用于各种组织和针对癌症进展的各种组织和疗法的受控方法的方法都非常重要。因此，该申请与国家卫生研究院的使命有很强的相关性。