Collaborative Research: Ideas Lab: The Role of Extracellular RNA in Intercellular and Interkingdom Communication

合作研究：创意实验室：细胞外 RNA 在细胞间和王国间通讯中的作用

• 批准号: 2243533
• 负责人: Kendall Corbin
• 金额: $ 85.63万
• 依托单位: University of Kentucky Research Foundation
• 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: 2023-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2243533&HistoricalAwards=false
• 关键词: Collaborative; Research; Ideas; Lab; Role

This project is a collaboration among seven laboratories with diverse and complementary expertise. The overarching goal of the project is to understand the role of extracellular RNA (exRNA) in communication between cells and in shaping the community of microbes, especially bacteria, that live on and inside plants, insects, and humans. These collections of microbes are often referred to as microbiomes and are critical for the health of plants and animals, including humans. A healthy microbiome promotes a healthy immune system, but how healthy microbiomes are maintained is poorly understood. This project will test the hypothesis that RNA secreted by host cells plays a central role in maintaining health both through communication among cells and by modifying the microbiome. RNA is best known for its key role in protein production inside cells, such as in RNA-based COVID vaccines. However, not all RNA encodes proteins, and cells produce more non-coding RNA than coding RNA, some of which is actively pushed into the environment by cells. This secreted RNA appears to be taken up by other cells, including bacteria and fungi, where it could potentially impact their growth. Understanding how exRNAs shape communication between cells and organisms will enable manipulation of exRNA communication in both agriculture and medicine, which will lead to development of environmentally friendly pesticides, as well as treatments that promote formation of healthy microbiomes in both plants and animals. This knowledge will also enable development of diagnostic and therapeutic tools for early detection and/or treatment of disease.All cellular organisms secrete RNAs. The functions of these extracellular RNAs (exRNAs), however, are poorly understood. Two likely functions are intercellular and interkingdom communication. Open questions abound in exRNA biology: how are exRNAs selected for secretion, how are they targeted to recipient cells, and what are their roles in normal health and organismal fitness? Arabidopsis leaf exRNA isolates are highly enriched in the post-transcriptional modification N6-methyladenosine (m6A) (as compared to total cellular RNA) suggesting that post-transcriptional modifications may tag specific RNAs for export. Consistent with this, human exosomal microRNAs are enriched with m6A (relative to cytosolic microRNAs). Interestingly, a large number of mammalian small non-coding RNAs (ncRNAs) that localize to the external cell surface were recently found to harbor specific sialylated glycan modifications. These observations suggest that specific RNA modifications tag RNAs for cellular export and direct entry into appropriate recipient cells. This project will 1) test the hypothesis that exRNAs have specific features marking them for secretion and uptake, both within and among species, 2) determine how exRNAs are transferred from signaler to receiver cells, and 3) assess the impact of exRNA on microbiome health and composition through examining human gut, insect gut, and leaf surface models.This project was co-funded by the Directorate for Biological Sciences, and the Plant Genome Research Program and the Plant Biotic Interactions Program in the Division of Integrative Organismal Systems.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.

该项目是七个具有多样化和互补专业知识的实验室之间的合作。该项目的首要目标是了解细胞外 RNA (exRNA) 在细胞间通讯和塑造微生物群落中的作用，特别是生活在植物、昆虫和人类体内的细菌。这些微生物集合通常被称为微生物组，对于包括人类在内的植物和动物的健康至关重要。健康的微生物组可以促进健康的免疫系统，但人们对如何维持健康的微生物组知之甚少。该项目将验证宿主细胞分泌的 RNA 通过细胞间通讯和改变微生物组在维持健康方面发挥核心作用的假设。 RNA 因其在细胞内蛋白质生产中的关键作用而闻名，例如在基于 RNA 的新冠疫苗中。然而，并非所有 RNA 都编码蛋白质，细胞产生的非编码 RNA 比编码 RNA 更多，其中一些被细胞主动推入环境中。这种分泌的 RNA 似乎被其他细胞吸收，包括细菌和真菌，可能会影响它们的生长。了解 exRNA 如何塑造细胞和生物体之间的通讯，将能够在农业和医学中操纵 exRNA 通讯，从而开发环境友好型农药，以及促进植物和动物中健康微生物组形成的治疗方法。这些知识还将有助于开发早期检测和/或治疗疾病的诊断和治疗工具。所有细胞生物体都会分泌 RNA。然而，人们对这些细胞外 RNA (exRNA) 的功能知之甚少。两个可能的功能是细胞间和界间通讯。 exRNA 生物学中存在大量悬而未决的问题：如何选择 exRNA 进行分泌，它们如何靶向受体细胞，以及它们在正常健康和机体健康中的作用是什么？拟南芥叶 exRNA 分离物高度富集转录后修饰 N6-甲基腺苷 (m6A)（与总细胞 RNA 相比），表明转录后修饰可能标记特定的 RNA 以供输出。与此一致的是，人外泌体 microRNA 富含 m6A（相对于胞质 microRNA）。有趣的是，最近发现大量定位于细胞外表面的哺乳动物小非编码 RNA (ncRNA) 具有特定的唾液酸化聚糖修饰。这些观察结果表明，特定的 RNA 修饰标记 RNA，用于细胞输出并直接进入适当的受体细胞。该项目将 1) 检验 exRNA 具有标记其在物种内部和物种间分泌和摄取的特定特征的假设，2) 确定 exRNA 如何从信号细胞转移到受体细胞，以及 3) 通过检查人类肠道、昆虫肠道和叶子表面模型来评估 exRNA 对微生物组健康和组成的影响。该项目由生物科学理事会、植物基因组研究计划和植物研究所共同资助 综合有机系统部门的生物相互作用计划。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。