Collaborative Research: Unraveling extracellular microRNA communication

这个项目的长期目标是更好地了解细胞之间的通信方式。这个项目的重点是通过microRNAs进行细胞间通信这一相对未被探索的过程。理解microRNA介导的细胞间通信的一个主要障碍是无法区分供体细胞和受体细胞产生的microRNA。该项目将采用一种创新的方法来绕过这一障碍。这项工作的成功完成将使人们更好地理解microRNA介导的细胞-细胞通信，并为研究包括人类在内的各种生物体的细胞-细胞相互作用提供一套新的资源和工具。microRNAs(MiRNAs)是一种在所有生理过程中都具有重要调节作用的非编码小RNA。MiRNAs被认为只在细胞内稳定，在细胞外被酶迅速降解。最近的研究表明情况并非如此：细胞可以通过多种机制输出miRNAs，影响隔壁或很远的其他细胞。过去的工作通常一次只关注一个miRNA，因为不可能区分一个细胞制造的miRNA分子和另一个细胞制造的miRNA分子。主要问题仍然没有得到回答：哪些miRNA被转移？细胞使用什么机制来转移它们？为了克服障碍并回答这些问题，将利用miRNA生物学、合成生物学和基因组编辑方面的专业知识。它将采用非哺乳动物系统的两个特征：标记RNA的原生动物酶，以及设计定制遗传电路和引入基因组修改的CRISPR/CA。将设计成标记RNA的细胞与那些不标记RNA的细胞相结合，将能够识别在细胞之间转移的miRNA物种。人类细胞中定制合成生物电路的实施和稳定集成将使对miRNA水平的可靠监测成为可能。选择性药物阻断细胞间通讯机制将有助于确定这种转移是如何发生的。基因组编辑可以精确地确定哪些基因参与了特定的转移机制，是细胞间miRNA交换的关键决定因素。长期目标是解决细胞之间如何相互通信的问题。拟议工作的成功完成将确定介导细胞间通信的miRNAs，并产生一套新的资源和工具，用于研究许多不同人类生物学模型中细胞之间的关系。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The long-term goal of this project is to better understand how cells communicate with each other. This project focuses on the relatively unexplored process of cell-cell communication via microRNAs. A major roadblock to understanding microRNA-mediated cell-cell communication has been the inability to distinguish microRNAs produced by donor and recipient cells. This project will employ an innovative approach to get around this roadblock. The successful completion of this work will lead to a better understanding of microRNA-mediated cell-cell communication and a new set of resources and tools for investigating cell-cell interactions in a variety of organisms, including humans.MicroRNAs (miRNAs) are small non-coding RNAs with significant regulatory roles in all physiological processes. miRNAs were thought to be stable only inside cells and rapidly degraded by enzymes when outside the cell. Recent work has demonstrated that this is not the case: cells can export miRNAs through multiple mechanisms, affecting other cells either next door or at great distances away. Past work has typically focused on one miRNA at a time because it is impossible to distinguish miRNA molecules made by one cell from those made by another. Major questions remain unanswered: Which miRNAs are transferred? What mechanisms do cells use to transfer them? To overcome hurdles and answer these questions, expertise in miRNA biology, synthetic biology and genome editing will be leveraged. Two features of non-mammalian systems will be adopted: a protozoan enzyme to label RNA, and CRISPR/Cas to engineer custom genetic circuits and introduce genome modifications. Combining cells engineered to label RNA with those that do not label RNA will enable the identification of miRNA species that are transferred between cells. Implementation and stable integration of custom synthetic biology circuits in human cells will permit the reliable monitoring of miRNA levels. Selective pharmacological blocking of inter-cellular communication mechanisms will facilitate the determination of how such transfer occurs. Genome editing will allow pinpointing which genes involved in specific transfer mechanisms are the critical determinants of miRNA exchange between cells. The long-term goal is to address questions about how cells communicate with each other. Successful completion of the proposed work will identify miRNAs that mediate cell-cell communication and generate a new set of resources and tools for investigating the relationship between cells in many different models of human biology.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.