Exosome kinetics in vivo: cell type and drug specific effects

项目摘要 外切体已被报道为疾病和药物反应的生物标志物。它们是运输囊泡， 由细胞分泌，并将microRNAs、mRNA和蛋白质运送到其他细胞；它们可以转运分子- 通过血液循环清除到其他局部组织或远处组织。外切体和靶标 细胞含有一些表面分子(例如凝集素蛋白)，这些分子似乎会影响细胞的速度(动力学)和位置 外切体摄取。这些递送说明会因疾病或药物治疗的影响而改变 结果是外切体摄取改变。因此，了解这些变化可能会阐明新的生物标志物 与疾病和药物反应有关。总的来说，这些数据表明关键的未知数是 1)是什么因素改变了外切体动力学；2)这些因素是如何改变递送指令的。在这个项目中， 我将专注于了解分泌细胞的变化如何改变分泌的外切体的动力学。 申请者的长期目标是改进外体生物标记物的合理使用，以预测疾病和 治疗反应。了解外切体动力学有助于优化外切体生物标志物VALIDA。 TION研究设计并可能导致阐明生物标记物关联的机制的研究。 中心假设是来自不同细胞类型的分泌外切体具有不同的外切体动力学和 疾病细胞和正常细胞的药物治疗改变了分泌的外切体的动力学。目标1将 测定不同细胞类型分泌的外切体在体内的动力学。目标2将决定 药物处理细胞外切体的体内动力学参数。在完成这些研究后，美联社- Plicant希望开发出能够识别血液中外切体的动力学参数的动力学模型。这位K08男士 Tored临床科学家研究职业发展奖详细介绍了一项为期4年的培训计划，旨在满足- PLISH 4个主要目标：(1)学习进行活体外切体动力学所需的技术和概念 研究和推进临床药理学领域；(2)培训高级药理学的概念和应用 生物信息学和外切体动力学参数的数据处理促进临床药理研究领域的发展 (3)获得将基本发现转化为合作的经验，以及(4)发展必要的专业技能- 萨里希望有一个成功而独立的学术生涯。印第安纳大学医学院 承诺通过培训年轻研究人员促进卫生研究的发展。 临床药理学分部资金充足，高度合作，拥有强大的研究项目 成人、儿科和产科药物遗传学、药物代谢和药物相互作用。申请人的列车- ING将包括导师和由NIH资助的具有专业知识的调查人员组成的教职员工咨询委员会 在动力学、生物信息学、基于生理学的计算建模和循环外显体生物学方面。通过 此次K08奖项的完成，我将开发出高质量的出版物，卓有成效的合作，独立 (R01)和合作研究资金，以及我的导师和教师顾问的反馈。

Project Summary Exosomes have been reported as biomarkers of disease and drug response. They are transport vesicles that are secreted from cells and deliver microRNAs, mRNA, and proteins to other cells; they can transport mole- cules to other local tissues, or to distant tissues through blood circulation. Both the exosomes and the target cells contain some surface molecules (e.g. lectin proteins) that appear to impact the rate (kinetics) and site of exosome uptake. These delivery instructions are changed by the effects of disease or drug treatment and re- sult in altered exosome uptake. Thus, understanding these changes may elucidate new biomarkers that are associated with both diseases and drug responses. Collectively, these data indicate that critical unknowns are 1) what factors alter exosome kinetics and 2) how do those factors alter the delivery instructions. In this project, I will focus on understanding how alterations to the secreting cells alter the kinetics of the secreted exosomes. The applicant's long-term goal is to improve the rational use of exosomal biomarkers to predict disease and treatment response. Understanding the exosome kinetics would help to optimize exosomal-biomarker valida- tion study designs and may lead to studies that elucidate the mechanisms underlying biomarker associations. The central hypothesis is that secreted exosomes from different cell types have different exosome kinetics and that the drug treatment of diseased and normal cells changes the kinetics of the secreted exosomes. Aim 1 will determine the in vivo exosome kinetics of exosomes secreted by different cell types. Aim 2 will determine the in vivo kinetic parameters of exosomes derived from drug treated cells. By completion of these studies, the ap- plicant expects to develop kinetic models that discern kinetic parameters of exosomes in blood. This K08 Men- tored Clinical Scientist Research Career Development Award details a 4-year training plan designed to accom- plish 4 main objectives: (1) learn the techniques and concepts required to conduct in vivo exosome kinetic studies and advance the field of clinical pharmacology, (2) train in the concepts and application of advanced bioinformatics and data processing of exosome kinetic parameters to advance the field of clinical pharmacolo- gy, (3) gain experience translating basic findings into collaborations, and (4) develop professional skills neces- sary for a successful and independent academic career. The Indiana University School of Medicine (IUSM) has made a commitment to promoting the growth of health research through the training of young investigators. The Division of Clinical Pharmacology is well-funded, highly collaborative, with strong research projects in adult, pediatric, and obstetric pharmacogenetics, drug metabolism, and drug interactions. The applicant's train- ing will include mentors and a faculty advisory committee consisting of NIH-funded investigators with expertise in kinetics, bioinformatics, physiologically-based computational modeling, and circulating exosome biology. By the completion of this K08 award, I will develop high-quality publications, fruitful collaborations, independent (R01) and collaborative research funding with feedback from my mentors and faculty advisors.