Doctoral Dissertation Research: Cold adaptation and gene expression

博士论文研究：冷适应与基因表达

• 批准号: 2316990
• 负责人: Christopher Schmitt
• 金额: $ 2.72万
• 依托单位: Trustees of Boston University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2316990&HistoricalAwards=false
• 关键词: Doctoral; Dissertation; Research; Cold; adaptation

In the past ~450,000 years, members of the genus Homo migrated multiple times from equatorial Africa to more seasonal and cold climates. Non-shivering thermogenesis – a process by which brown fat generates heat – was likely a key part of the success of our genus in adapting to these environments and helping to facilitate these geographic expansions. The product of the gene UCP1, a protein called thermogenin, drives this process. Nonhuman primates have also expanded from Africa into colder latitudes over the past ~300,000 years. Previously, the investigators found signals of selection in the regulation of this gene in some nonhuman primates. In this doctoral dissertation research project, the investigators validate and expand those findings by using innovative methods and previously collected samples. Students from groups underrepresented in STEM disciplines receive training and professional development through this project.Using an established protocol newly validated for specific species, the investigators chemically convert archived wild nonhuman primate fibroblast cell lines into chemically induced brown adipose (ciBA) cells. Outcomes of the conversion are verified by quantifying RNA expression of UCP1 (which is uniquely expressed in brown adipose) via RT-qPCR, with Oil Red O staining and immunofluorescence staining to observe lipid droplet formation and the completeness of culture conversion. The ciBA are then exposed to norepinephrine to stimulate a cold response in the cell lines, and UCP1 gene expression with and without the cold response are compared using bulk RNA sequencing. Results are used to test the prediction of a derived allele dosage effect in UCP1 expression across four loci, such that each derived allele, previously found to have appeared at higher frequencies in colder climates, will be associated with a significant increase in UCP1 expression. Analyses consider potential covariates like sex, BMI, and body weight. Validation of the investigators previous results suggest that the NST system has undergone positive selection in response to colder climates in nonhuman primates and inform our understanding of this process in our own genus.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在过去的〜450，000年中，Homo属的成员多次从非洲的赤道迁移到季节性和寒冷的气候。非动摇的生热发生 - 棕色脂肪产生热量的过程 - 可能是我们属在适应这些环境并有助于促进这些地理扩张的成功中成功的关键部分。该基因UCP1的乘积是一种称为热生蛋白的蛋白质，可驱动这一过程。在过去的30万年中，非人类灵长类动物也从非洲扩展为较冷的纬度。以前，研究人员发现了一些非人类私人中该基因调节中选择的信号。在这个博士学位论文研究项目中，研究人员通过使用创新方法和先前收集的样本来验证和扩展这些发现。来自STEM学科中代表性不足的群体的学生通过该项目接受培训和专业发展。使用针对特定物种的既定方案，研究人员化学转化的存档的野生非人类灵长类动物成纤维细胞系进入化学诱导的棕色脂肪（CIBA）细胞。通过量化UCP1的RNA表达（在棕色脂肪中唯一表达）通过RT-QPCR来验证转化率的结果，并具有油红O染色和免疫荧光染色，以观察脂质液滴的形成以及培养转化的完整性。然后将CIBA暴露于去甲肾上腺素以刺激细胞系中的冷反应，并使用大量RNA测序比较带有冷反应的UCP1基因表达。结果用于测试四个基因座UCP1表达中衍生等位基因剂量效应的预测，因此每个衍生的等位基因先前发现在寒冷气候下以较高的频率出现，将与UCP1表达的显着增加有关。分析考虑性，BMI和体重等潜在的协变量。对研究人员的验证先前的结果表明，NST系统已经理解了对非人类隐私的寒冷气候的积极选择，并在我们自己的属中了解了我们对这一过程的理解。该奖项反映了NSF的法定任务，并被认为是通过基金会的知识分子优点和更广泛的影响审查标准来通过评估来获得的支持。