Mechanisms of developmental buffering

发育缓冲机制

• 批准号: 10556357
• 负责人: James Tucker Nichols
• 金额: $ 45.61万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10556357
• 关键词: Address; Adult; Affect; Animals; Breeding; Buffers; Cells; Counseling; Craniofacial Abnormalities; DNA; DNA Sequence; DNA Sequence Alteration; Defect; Dental; Development; Developmental Process; Disease; Disease Management; Dorsal; Epigenetic Process; Equilibrium; Experimental Designs; Fingers; Fishes; Future; Gene Expression; Gene Expression Regulation; Gene Family; Genes; Genetic; Genetic Diseases; Genetic Variation; Genotype; Head; Health; Heritability; Human; Human Genetics; Human Genome; Individual; Inherited; Label; Measurement; Measures; Medical; Methods; Mission; Modeling; Mutation; National Institute of Dental and Craniofacial Research; Oral; Organism; Orthologous Gene; Pathway interactions; Penetrance; Phenotype; Reiterated Genes; Reporting; Research; Signal Transduction; Skeleton; Specific qualifier value; Statistical Data Interpretation; Symptoms; System; Testing; Tissues; Variant; Work; Zebrafish; craniofacial; disease diagnosis; disease phenotype; disorder prevention; epigenome; epigenomic profiling; epigenomics; falls; flexibility; gene interaction; genetic manipulation; genetic predictors; genetic variant; genome editing; genome sequencing; improved; member; mutant; myocyte-specific enhancer-binding-factor 2C; notch protein; novel therapeutic intervention; response; screening; skeletal; transcription factor; transcriptomic profiling; zebrafish genome

PROJECT SUMMARY/ABSTRACT: The long-term objective of this proposal is to understand genetic buffering, or how some individuals overcome the effects of a harmful genetic mutation. We generated a zebrafish model of buffering in order to understand how individuals who should have gotten sick are somehow able to live unaffected healthy lives. In our model, we used selective breeding to generate two strains of zebrafish that develop dramatically different head skeletons in response to the same harmful genetic mutation. In one strain, the mutation causes severe, lethal skeletal defects. Meanwhile, the other strain is remarkably buffered against the mutation. The buffered fish develop essentially normal head skeletons, surviving to be fertile, viable adults. We compare these strains to understand the natural buffering mechanisms that are present in some fish, and likely in some humans too. We hypothesize that buffering is due to factors that tune developmental processes to restore balance. For example, in Aim1 we will determine how DNA sequences which oppose the mutant gene can be turned down. In Aim 2 we examine how factors encoded in the DNA which perform the same function as the mutant gene can be turned up. In Aim 3 we will determine how changes that do not necessarily involve alterations in the DNA sequence can buffer the harmful mutation. These three specific aims test how interacting mechanisms function together to buffer development. We designed experiments to address these aims using state of the art methods like genome editing and sequencing, colorful cell and tissue labeling, quantitative measurements of large numbers of fish skeletons, and rigorous statistical analyses. The mechanisms we propose to study here are present in many developmental systems and organisms and therefore will likely be applicable in wide- ranging settings. This study of buffering mechanisms could lead to novel therapeutic approaches, buffering mechanisms might be manipulated in the future to manage disease symptoms. This work will also lead to a better understanding of the factors that make predicting genetic disease from gene sequences difficult. Thus, this line of research will potentially inform and improve medical practices, including genetic disease management, disease diagnosis, and counseling, falling squarely within the mission of the NIDCR to improve dental, oral and craniofacial health through research.

项目概要/摘要： 该提案的长期目标是了解遗传缓冲，或者某些个体如何克服 有害基因突变的影响。我们生成了一个斑马鱼缓冲模型以便理解 本来应该生病的人如何能够过上不受影响的健康生活。在我们的模型中， 我们通过选择性育种培育了两种斑马鱼，它们的头部发育截然不同 骨骼对相同的有害基因突变做出反应。在一种菌株中，突变会导致严重的、致命的 骨骼缺陷。与此同时，另一种菌株对突变有显着的缓冲作用。缓冲的鱼 发育出基本正常的头部骨骼，存活下来成为具有生育能力的、有活力的成年人。我们将这些菌株与 了解某些鱼类以及某些人类中可能存在的自然缓冲机制。我们 假设缓冲是由于调整发育过程以恢复平衡的因素造成的。为了 例如，在 Aim1 中，我们将确定如何关闭对抗突变基因的 DNA 序列。 在目标 2 中，我们研究 DNA 中编码的因子如何执行与突变基因相同的功能 可以打开。在目标 3 中，我们将确定如何进行不一定涉及变更的变更 DNA序列可以缓冲有害的突变。这三个具体目标测试了如何相互作用的机制 共同发挥作用以缓冲发展。我们设计了实验来利用最先进的技术来实现这些目标 基因组编辑和测序、彩色细胞和组织标记、定量测量等方法 大量的鱼骨骼和严格的统计分析。我们建议在这里研究的机制 存在于许多发育系统和生物体中，因此可能适用于广泛的 测距设置。这项对缓冲机制的研究可能会带来新的治疗方法，缓冲 未来可能会操纵机制来控制疾病症状。这项工作也将导致 更好地了解导致从基因序列预测遗传病变得困难的因素。因此， 这一系列的研究将有可能为包括遗传疾病在内的医疗实践提供信息并改善医疗实践 管理、疾病诊断和咨询，完全属于 NIDCR 的使命，以改善 通过研究促进牙齿、口腔和颅面健康。