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高友鹤 北京师范大学 生命科学学院 Urimarker 2015 迎接尿标志物时代的到来 高友鹤 北京师范大学 生命科学学院 Urimarker 2015

为什么说现在是尿标志物研究的历史机遇？

前人没有想到吗？

Urinalysis 4000 B.C. Third milestone in the history of medicine

Physiologist JA Armstrong "From a liquid window through which physicians felt they could view the body's inner workings, urine led to the beginnings of laboratory medicine."

"As the role of physicians became elevated, the importance of urinary diagnosis became exaggerated (17th century), the uses of uroscopy had spiraled far beyond the edge of reason."

当前的状况

Urine/Blood in Biomarker Studies pubmed - (((blood) OR serum) OR plasma) AND biomarker pubmed -  (urine) AND biomarker year count urine/blood 2013 3436 302 8.79% 2012 20759 1401 6.75% 2011 19996 1369 6.85% 2010 19097 1288 6.74% 2009 17733 1056 5.95% 2008 16993 1057 6.22% 2007 16509 952 5.77% 2006 15186 918 6.05% 2005 15389 860 5.59% 2004 14208 767 5.40% 2003 13417 696 5.19% Up to 2013-5-29

什么原因尿液的研究不再受到重视？

Individual differences

基线都不平， 我们找错地方了？

我们想找的标志物是什么呢？

My Definition Biomarker is the measurable change associated with a physiological or pathophysiological process. blog.sciencenet.cn/home.php?mod=space&uid=244733&do=blog&id=725226

Biomarker=Change

Homeostasis mechanisms =minimal changes =less biomarkers In Blood Homeostasis mechanisms =minimal changes =less biomarkers

Accumulate all the changes = Lots of biomarkers In Urine Accumulate all the changes = Lots of biomarkers

Change removal Several organs are important in removing waste from the body. The lungs remove carbon dioxide. The liver converts excess protein into urea. The skin provides a surface for small amounts of water and salt to move out of the body. The kidneys remove unwanted substances such as urea, excess water and salt. Boardworks Ltd 2009

Changes of proteins induced by anticoagulants can be more sensitively detected in urine rather than plasma 李梦林明天报告

Individual differences

尿液能刻画复杂的生理状况 吴建强明天报告

生理状态下基本不变的， 如果发生了变化， 可以作为潜在疾病标志物。 2009 Proteomics Clinical Application

尿液也应该能刻画出早期微小的病理生理学变化

血、尿标志物比较的几个例子

Tianfu Wu et al @ UT Southwestern Medical Center Molecular & Cellular Proteomics (2013) 12:1170–1179

Payne, SR et al @ Seattle and Germany Prostate (2009) 69: 1257–1269

Not just within urinary tract This is the first study to demonstrate that analysis of urinary MMPs may be useful in determining disease status in a variety of human cancers, both within and outside of the urinary tract. MMP: 72KD 92KD >150KD Moses MA et al @ Harvard Med School CANCER RESEARCH (1998) 58, 1395-1399

Plasma and Urinary Desmosine as biomarkers for COPD Huang J et al@ UK Thorax (2012) 67: 502-508

Smith ER et al. @ Harvard Med School Clin Cancer Res 2008;14:2378-2386 Urine of brain tumor patients contains significantly higher levels of MMP-2, MMP-9, and MMP-9/NGAL compared with control samples. Urine of brain tumor patients contains significantly higher levels of MMP-2, MMP-9, and MMP-9/NGAL compared with control samples. Smith ER et al. @ Harvard Med School Clin Cancer Res 2008;14:2378-2386 ©2008 by American Association for Cancer Research

Smith ER et al. @ Harvard Med School Clin Cancer Res 2008;14:2378-2386 The loss of urinary MMPs after resection of a brain tumor, demonstrating that tumor presence is related to increased urinary MMP activity and removal of that tumor correlates with subsequent clearing of detectable urinary MMP activity. The loss of urinary MMPs after resection of a brain tumor, demonstrating that tumor presence is related to increased urinary MMP activity and removal of that tumor correlates with subsequent clearing of detectable urinary MMP activity. A, urine from a patient with a brain tumor (preop) contains MMP-2 and MMP-9 with strong zones of clearance indicating enzyme activity in the urine when the tumor is present (as seen in the preoperative magnetic resonance imaging; B). One year after a gross total resection of the tumor, with no residual/recurrent disease seen on the 1 y follow-up scan, the urinary zymogram reveals no observable MMP activity. The tumor of this patient is shown in the top axial T1 postgadolinium magnetic resonance imaging (B; arrow) with the 1 y follow-up scan next to (C), showing no tumor in the resection cavity at the time that the postoperative urine was analyzed. Smith ER et al. @ Harvard Med School Clin Cancer Res 2008;14:2378-2386 ©2008 by American Association for Cancer Research

Before Cerebral Ischemia in Rats 安满霞明天报告 Luigi Sironi et al @Italy Stroke. 2001;32:753-760.

尿液标志物的难点

Disadvantages as Biomarker Source Big variation Need more cases to validate

Way to solve the problem More sample Higher throughput MS More analyzing power

Problem with samples Very diluted Protein degradation

( Urimem

Urinary Proteins on Membrane

Changed the face of medicine “Hard as it is to believe today, a single concept developed by Dr. Henry Plummer  (1874 – 1936) at the beginning of the 20th century changed the face of medicine.” http://www.mayoclinic.org/tradition-heritage/medical-records.html

Changed the face of medicine The concept was a centralized medical record, stored in a single repository, and capable of traveling with the patient. (1907) Now paperless filmless chartless http://www.mayoclinic.org/tradition-heritage/medical-records.html

Biobanking’s future: Long-term at room temperature 张繁霜明天报告

Risk You Don’t Want to Take Others Discovery and validation in Blood Hard and expensive Patent Take your clue, validate in urine Easy and Cheap New patent

Tianfu Wu et al @ UT Southwestern Medical Center Molecular & Cellular Proteomics (2013) 12:1170–1179

Roadmap Minimizing confounding factors Using animal models (of disease) to study the effects in urine （袁媛明天报告） Validate in human samples

要考虑药物的影响 李逊斗明天报告

Urine smells good!

It tastes even better!

男怕入错行！

女更怕！

博士后招聘中

孙伟 各种体液的比较 邵晨 从尿液标志物数据库看尿生物标志物研究现状 李梦林 引入血液的变化在尿液中的反映 李逊斗 药物对尿蛋白质组的影响 赵敏迪 尿蛋白质组学的现状及尿蛋白质组的个体差异 贾露露 寻找器官特异标志物的一个策略 袁媛 肾脏疾病动物模型的尿蛋白质组 张繁霜 尿液的蛋白和核酸的膜保存 秦伟伟 膜上蛋白的洗脱方法 吴建强 尿液对生理功能的反映 安满霞 尿液对脑部疾病的反映

Acknowledgement Menglin Li Lulu Jia Liu Liu Department of Nephrology, PUMC Hospital: Mingxi Li, Xuejiao Liu Funding from Project 973

Thanks! Youhe Gao (高友鹤) 科学网博客，Linkedin gaoyouhe@bnu.edu.cn