The Platelet Transcriptome and Organ Failure After Injury: Discovering Molecular Biomarkers and Preventative Targets through Interrogating Novel RNA Modifications

项目摘要/摘要 作为一名创伤外科医生-科学家和早期研究人员，在NIGMS指导职业生涯的第五年 损伤后血小板功能的翻译研究发展奖，我发展了专业知识， 基础设施，并在证明创伤后早期血小板功能障碍方面做出了重大贡献 常见的，由生理变化驱动的，并与后来的器官衰竭发展有关。这是 这很重要，因为伤害仍然是世界范围内主要的死亡原因，但初步护理的进展已经发生了变化 损伤相关的发病率和死亡率的负担从损伤后早期到后期，留下了两个重要的 挑战：1)确定最初和持续器官衰竭风险最高的受伤幸存者 导致长期发病率和晚期死亡率，以及2)发现早期治疗的新靶点以预防 损伤后器官衰竭的发展。 在血栓性炎症中，血小板通过病理生物学作用导致器官衰竭。在类似的疾病中，血小板 转录学提高了对机制的理解，并推动了对基于血小板的探索 治疗学。血小板巨核细胞衍生的核糖核酸(RNAs)在健康中是稳定的，但被 疾病中的生理信号。重要的是，血小板是无核的，缺乏RNA合成，因此提供了 人类生物学中关于RNA修饰的最干净的转录切割学观点。因此，我的建议旨在重新- 想象一下，我们是如何通过专注于 用血小板转录组探讨早期患者器官衰竭的预测和预防 存活下来，并确定可以在模型系统中测试的RNA修饰靶点 解决这些知识差距：1)确定严重损伤和 在纵向患者研究中，它与器官衰竭的发展和解决的关系；2)定义 血小板RNA修饰对严重创伤患者和急性期患者血小板功能的影响 体内修饰的健康血小板；3)开发体外和体外模型系统，以操纵临床相关 血小板RNA修饰作为体内模型和临床测试的最终通道。我将使用可行性(下一步 世代测序、核糖体足迹分析)和新颖(亚群测序、体外转基因 血小板模型系统)识别新的分子生物标记物和预防靶点的方法 损伤后器官衰竭的发生发展，并创新我们对生理驱动的RNA的理解 在最佳急性生理变化下通过理想的血小板无核生物学进行修饰 受伤。这项从K奖到R奖的过渡建议是NIGMS的任务重点，通过使用基础研究来提高 了解生物过程，并为疾病诊断、治疗和治疗的进展奠定基础 以及在“伤害”范围内的预防(药理学和生理学)。

Project Summary/Abstract As a trauma surgeon-scientist and early stage investigator in the 5th year of an NIGMS mentored career development award for the translational study of post-injury platelet function, I have developed expertise, infrastructure, and made significant contributions to demonstrating that early platelet dysfunction after injury is common, driven by changing physiology, and associated with later development of organ failure. This is important because injury remains a leading cause of death worldwide, but advances in initial care have shifted the burden of injury-related morbidity and mortality from early after injury to later, leaving two important challenges:1) identifying injured survivors at highest risk of developing initial and sustained organ failure leading to long-term morbidity and late mortality, and 2) discovering novel targets for early therapies to prevent development of organ failure after injury. Platelets contribute to organ failure through pathobiology in thromboinflammation. In similar diseases, platelet transcriptomics has improved mechanistic understandings and driven exploration of platelet-based therapeutics. Platelet megakaryocyte derived ribonucleic acids (RNAs) are stable in health, but modified by physiological signals in disease. Importantly, platelets are anucleate and lack RNA synthesis, thus providing the cleanest transcriptomic view of RNA modification in human biology. As such, my proposal seeks to re- imagine how we understand, measure, and intervene on early alterations in platelets after injury by focusing on the platelet transcriptome to explore prediction and prevention of organ failure among patients who initially survive their injuries, and to identify RNA modification targets that can be tested in model systems by addressing these knowledge gaps: 1) Identify the early platelet transcriptional landscape of severe injury and its relationship to development and resolution of organ failure in longitudinal patient studies; 2) Define the effect of platelet RNA modifications on platelet function in samples from patients with severe injury and in ex vivo modified healthy platelets; 3) Develop ex vivo and in vitro model systems to manipulate clinically relevant platelet RNA modifications as ultimate conduits to in vivo models and clinical testing. I will use feasible (next generation sequencing, ribosome footprint profiling), and novel (sub-population sequencing, ex vivo transgenic platelet model systems) methods to identify novel molecular biomarkers and preventative targets of development of organ failure after injury, and innovate our understanding of physiologically driven RNA modification through the ideal anucleate biology of platelets under the optimal acute physiologic changes of injury. This K to R award transition proposal is NIGMS mission-focused by using basic research to increase understanding of biological processes and lay the foundation for advances in disease diagnosis, treatment, and prevention within the area of ‘Injury’ (Pharmacological and Physiological Sciences Branch).