Author:
Lehmann-Werman Roni,Neiman Daniel,Zemmour Hai,Moss Joshua,Magenheim Judith,Vaknin-Dembinsky Adi,Rubertsson Sten,Nellgård Bengt,Blennow Kaj,Zetterberg Henrik,Spalding Kirsty,Haller Michael J.,Wasserfall Clive H.,Schatz Desmond A.,Greenbaum Carla J.,Dorrell Craig,Grompe Markus,Zick Aviad,Hubert Ayala,Maoz Myriam,Fendrich Volker,Bartsch Detlef K.,Golan Talia,Ben Sasson Shmuel A.,Zamir Gideon,Razin Aharon,Cedar Howard,Shapiro A. M. James,Glaser Benjamin,Shemer Ruth,Dor Yuval
Abstract
Minimally invasive detection of cell death could prove an invaluable resource in many physiologic and pathologic situations. Cell-free circulating DNA (cfDNA) released from dying cells is emerging as a diagnostic tool for monitoring cancer dynamics and graft failure. However, existing methods rely on differences in DNA sequences in source tissues, so that cell death cannot be identified in tissues with a normal genome. We developed a method of detecting tissue-specific cell death in humans based on tissue-specific methylation patterns in cfDNA. We interrogated tissue-specific methylome databases to identify cell type-specific DNA methylation signatures and developed a method to detect these signatures in mixed DNA samples. We isolated cfDNA from plasma or serum of donors, treated the cfDNA with bisulfite, PCR-amplified the cfDNA, and sequenced it to quantify cfDNA carrying the methylation markers of the cell type of interest. Pancreatic β-cell DNA was identified in the circulation of patients with recently diagnosed type-1 diabetes and islet-graft recipients; oligodendrocyte DNA was identified in patients with relapsing multiple sclerosis; neuronal/glial DNA was identified in patients after traumatic brain injury or cardiac arrest; and exocrine pancreas DNA was identified in patients with pancreatic cancer or pancreatitis. This proof-of-concept study demonstrates that the tissue origins of cfDNA and thus the rate of death of specific cell types can be determined in humans. The approach can be adapted to identify cfDNA derived from any cell type in the body, offering a minimally invasive window for diagnosing and monitoring a broad spectrum of human pathologies as well as providing a better understanding of normal tissue dynamics.
Funder
Juvenile Diabetes Research Foundation International
HHS | NIH | National Institute of Diabetes and Digestive and Kidney Diseases
Israel Science Foundation
Sir Zalman Cowen Universities Fund
Deutsche Forschungsgemeinschaft
Sokya Pancreatic cancer research program
Teva Pharmaceutical Industries
Publisher
Proceedings of the National Academy of Sciences
Cited by
477 articles.
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