Collaborative Research: Role of Endothelial Cell Activation in Hypoxia Tolerance of an Elite Diver, the Weddell Seal

合作研究：内皮细胞激活在精英潜水员威德尔海豹耐缺氧中的作用

• 批准号: 2020706
• 负责人: Allyson Hindle
• 金额: $ 56.88万
• 依托单位: University of Nevada Las Vegas
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2020706&HistoricalAwards=false
• 关键词: Collaborative; Research; Role; Endothelial; Cell

This award is funded in whole or in part under the American Rescue Plan Act of 2021 (Public Law 117-2). Part I: Non-technical description: The Weddell seal is an iconic Antarctic species and a superb diver, swimming down to 2,000 feet and staying underwater for up to 45 minutes. However, as for any mammal, the low oxygen concentrations in the blood during diving and the recovery once back at the surface are challenges that need to be overcome making their diving ability something unique that has fascinated scientists for decades. This research project will evaluate the underlying processes in Weddell seal’s physiology that protects this species from the consequences of diving. The work will combine laboratory experiments where cells that line the blood vessels will be exposed to conditions of low oxygen, similar to those that will be measured in diving seals in Antarctica. The investigarors will test a new idea that several short-term dives, performed before a long dive, allows seals to condition themselves. Measurements on the chemical compounds released to the blood during dives, combined with experiments on the genes that regulate them will provide clues on the biochemical pathways that help the seals tolerate these extreme conditions. The project allows for documentation of individual seal dives and provisioning of such information to the broader science community that seeks to study these seals, educating graduate and undergraduate students and a post-doctoral researcher and producing a science-outreach comic book for middle-school students to illustrate the project's science activities, goals and outcomes. Part II: Technical description: The Weddell seal is a champion diver with high natural tolerance for low blood oxygen concentration (hypoxemia) and inadequate blood supply (ischemia). The processes unique to this species protects their tissues from inflammation and oxidative stress observed in other mammalian tissues exposed to such physiological conditions. This project aims to understand the signatures of the processes that protect seals from inflammation and oxidant stress, using molecular, cellular and metabolic tools. Repetitive short dives before long ones are hypothesized to precondition seal tissues and activate the protective processes. The new aspect of this work is the study of endothelial cells, which sense changes in oxygen and blood flow, providing a link between breath-holding and cellular function. The approach is one of laboratory experiments combined with 2-years of field work in an ice camp off McMurdo Station in Antarctica. The study is structured by three main objectives: 1) laboratory experiments with arterial endothelial cells exposed to changes in oxygen and flow to identify molecular pathways responsible for tolerance of hypoxia and ischemia using several physiological, biochemical and genomic tools including CRSPR/Cas9 knochout and knockdown approaches. 2) Metabolomic analyses of blood metabolites produced by seals during long dives. And 3) Metabolomic and genomic determinations of seal physiology during short dives hypothesized to pre-condition tolerance responses. In the field, blood samples will be taken after seals dive in an isolated ice hole and its diving performance recorded. It is expected that the blood will contain metabolites that can be related to molecular pathways identified in lab experiments. Expert collaborators will provide field support, with the ice camp, dive hole for the seals, and telemetry associated with the seals’ dives. The project builds upon previous NSF-funded projects where the seal genome and cellular resources were produced. Undergraduate researchers will be recruited from institutional programs with a track record of attracting underrepresented minorities and a minority-serving institution. To further increase polar literacy training and educational impacts, the field team will include a blog where field experiences are shared and comic book preparation with an artist designed for K-12 students and public outreach.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.

该奖项是根据2021年《美国救援计划法》（公法117-2）全部或部分资助的。第一部分：非技术描述：Weddell Seal是一种标志性的南极物种，是一位出色的潜水员，游泳至2,000英尺，在水下停留长达45分钟。但是，至于任何哺乳动物，潜水期间血液中的氧气浓度低，而恢复后的恢复是需要克服的挑战，这使他们的潜水能力使他们的潜水能力变得独特，使科学家着迷了数十年。该研究项目将评估Weddell Seal生理学中的基本过程，该过程保护该物种免受潜水后果的影响。这项工作将结合实验室实验，在该实验实验中，将血管围绕低氧条件的细胞与在南极洲潜水密封中测量的细胞相似。研究人员将测试一个新想法，即在长时间潜水之前进行几次短期潜水，可以使密封能够调节自己。在潜水过程中释放给血液的化合物的测量，结合对其调节它们的基因的实验，将为生化途径提供线索，以帮助密封剂耐受这些极端条件。该项目允许记录单个密封潜水并向更广泛的科学界提供此类信息，该社区旨在研究这些印章，教育研究生和本科生以及博士后研究人员，并为中学学生制作一本科学 - 偶像漫画书，以说明项目的科学活动，目标和成果。第二部分：技术描述：Weddell密封是一名冠军潜水员，对低血氧浓度（低氧血症）和血液供应不足（缺血）具有很高的自然耐受性。该物种独有的过程可以保护其组织免受炎症和在暴露于这种物理条件的其他哺乳动物组织中观察到的炎症和氧化应激。该项目旨在了解使用分子，细胞和代谢工具来保护密封免受注射和氧化物应力的过程的特征。重复的短潜水在长时间的短潜水中被假设到前提密封组织并激活保护过程。这项工作的新方面是对内皮细胞的研究，这些细胞感知氧气和血流的变化，从而提供了呼吸持续和细胞功能之间的联系。该方法是实验室实验之一，结合了南极洲麦克默多车站（McMurdo Station）附近的冰训练营的2年野外工作。该研究由三个主要目标结构：1）实验室实验的动脉内皮细胞暴露于氧气和流量变化的动脉内皮细胞，以鉴定使用几种物理，生化和基因组的耐受性低氧和缺血的分子途径。包括CRSPR/CAS9 Knochout和敲低方法在内的工具。 2）长潜水过程中密封产生的血液代谢产物的代谢组分析。 3）在短潜水中对海豹生理的代谢组和基因组测定，假设条件前耐受性反应。在该领域，将在海豹潜水进入一个孤立的冰洞并记录其潜水性能之后采集血液样本。预计血液将含有可能与实验实验中鉴定的分子途径有关的代谢产物。专家合作者将提供现场支持，并提供冰训练营，海豹的潜水洞以及与海豹突然潜水有关的遥测。该项目建立在先前由NSF资助的项目上，在该项目中生产了密封基因组和细胞资源。本科研究人员将从具有吸引代表性不足的少数民族和少数族裔服务机构的往绩中招募。为了进一步增加极地识字培训和教育影响，现场团队将包括一个博客，与专为K-12学生和公众推广设计的艺术家共享现场经验并准备漫画书。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。