Fragment Length of Circulating Tumor DNA

Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor recurrence, and evaluate response to therapy solely through a non-invasive blood draw. However, detecting ctDNA against the normally occurring back...

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Vydáno v:	PLoS genetics Ročník 12; číslo 7; s. e1006162
Hlavní autoři:	Underhill, Hunter R., Kitzman, Jacob O., Hellwig, Sabine, Welker, Noah C., Daza, Riza, Baker, Daniel N., Gligorich, Keith M., Rostomily, Robert C., Bronner, Mary P., Shendure, Jay
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	United States Public Library of Science 01.07.2016 Public Library of Science (PLoS)
Témata:	Alleles Animals Biology and life sciences Biomarkers, Tumor - blood Carcinoma, Hepatocellular - genetics Carcinoma, Hepatocellular - metabolism Cell Line, Tumor Deoxyribonucleic acid DNA DNA sequencing DNA, Neoplasm - blood DNA, Neoplasm - genetics Experiments Genes Genetic aspects Glioblastoma - blood Glioblastoma - genetics Hep G2 Cells Humans Liver Neoplasms - genetics Liver Neoplasms - metabolism Lung cancer Lung Neoplasms - genetics Lung Neoplasms - metabolism Magnetic Resonance Imaging Male Medicine and Health Sciences Melanoma Melanoma - genetics Melanoma - metabolism Metastasis Methods Mutation Neoplasm Transplantation Proto-Oncogene Proteins B-raf - genetics Rats Research and Analysis Methods Studies Tumors
ISSN:	1553-7404, 1553-7390, 1553-7404
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Abstract	Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor recurrence, and evaluate response to therapy solely through a non-invasive blood draw. However, detecting ctDNA against the normally occurring background of cell-free DNA derived from healthy cells has proven challenging, particularly in non-metastatic solid tumors. In this study, distinct differences in fragment length size between ctDNAs and normal cell-free DNA are defined. Human ctDNA in rat plasma derived from human glioblastoma multiforme stem-like cells in the rat brain and human hepatocellular carcinoma in the rat flank were found to have a shorter principal fragment length than the background rat cell-free DNA (134-144 bp vs. 167 bp, respectively). Subsequently, a similar shift in the fragment length of ctDNA in humans with melanoma and lung cancer was identified compared to healthy controls. Comparison of fragment lengths from cell-free DNA between a melanoma patient and healthy controls found that the BRAF V600E mutant allele occurred more commonly at a shorter fragment length than the fragment length of the wild-type allele (132-145 bp vs. 165 bp, respectively). Moreover, size-selecting for shorter cell-free DNA fragment lengths substantially increased the EGFR T790M mutant allele frequency in human lung cancer. These findings provide compelling evidence that experimental or bioinformatic isolation of a specific subset of fragment lengths from cell-free DNA may improve detection of ctDNA.
AbstractList	Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor recurrence, and evaluate response to therapy solely through a non-invasive blood draw. However, detecting ctDNA against the normally occurring background of cell-free DNA derived from healthy cells has proven challenging, particularly in non-metastatic solid tumors. In this study, distinct differences in fragment length size between ctDNAs and normal cell-free DNA are defined. Human ctDNA in rat plasma derived from human glioblastoma multiforme stem-like cells in the rat brain and human hepatocellular carcinoma in the rat flank were found to have a shorter principal fragment length than the background rat cell-free DNA (134–144 bp vs. 167 bp, respectively). Subsequently, a similar shift in the fragment length of ctDNA in humans with melanoma and lung cancer was identified compared to healthy controls. Comparison of fragment lengths from cell-free DNA between a melanoma patient and healthy controls found that the BRAF V600E mutant allele occurred more commonly at a shorter fragment length than the fragment length of the wild-type allele (132–145 bp vs. 165 bp, respectively). Moreover, size-selecting for shorter cell-free DNA fragment lengths substantially increased the EGFR T790M mutant allele frequency in human lung cancer. These findings provide compelling evidence that experimental or bioinformatic isolation of a specific subset of fragment lengths from cell-free DNA may improve detection of ctDNA. During cell death, DNA that is not contained within a membrane (i.e., cell-free DNA) enters the circulation. Detecting cell-free DNA originating from solid tumors (i.e., circulating tumor DNA, ctDNA), particularly solid tumors that have not metastasized, has proven challenging due to the relatively abundant background of normally occurring cell-free DNA derived from healthy cells. Our study defines the subtle but distinct differences in fragment length between normal cell-free DNA and ctDNA from a variety of solid tumors. Specifically, ctDNA was overall consistently shorter than the fragment length of normal cell-free DNA. Subsequently, we showed that a size-selection for shorter cell-free DNA fragments increased the proportion of ctDNA within a sample. These results provide compelling evidence that development of techniques to isolate a subset of cell-free DNA consistent with the ctDNA fragment lengths described in our study may substantially improve detection of non-metastatic solid tumors. As such, our findings may have a direct impact on the clinical utility of ctDNA for the non-invasive detection and diagnosis of solid tumors (i.e., the “liquid biopsy”), monitoring tumor recurrence, and evaluating tumor response to therapy. Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor recurrence, and evaluate response to therapy solely through a non-invasive blood draw. However, detecting ctDNA against the normally occurring background of cell-free DNA derived from healthy cells has proven challenging, particularly in non-metastatic solid tumors. In this study, distinct differences in fragment length size between ctDNAs and normal cell-free DNA are defined. Human ctDNA in rat plasma derived from human glioblastoma multiforme stem-like cells in the rat brain and human hepatocellular carcinoma in the rat flank were found to have a shorter principal fragment length than the background rat cell-free DNA (134-144 bp vs. 167 bp, respectively). Subsequently, a similar shift in the fragment length of ctDNA in humans with melanoma and lung cancer was identified compared to healthy controls. Comparison of fragment lengths from cell-free DNA between a melanoma patient and healthy controls found that the BRAF V600E mutant allele occurred more commonly at a shorter fragment length than the fragment length of the wild-type allele (132-145 bp vs. 165 bp, respectively). Moreover, size-selecting for shorter cell-free DNA fragment lengths substantially increased the EGFR T790M mutant allele frequency in human lung cancer. These findings provide compelling evidence that experimental or bioinformatic isolation of a specific subset of fragment lengths from cell-free DNA may improve detection of ctDNA. Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor recurrence, and evaluate response to therapy solely through a non-invasive blood draw. However, detecting ctDNA against the normally occurring background of cell-free DNA derived from healthy cells has proven challenging, particularly in non-metastatic solid tumors. In this study, distinct differences in fragment length size between ctDNAs and normal cell-free DNA are defined. Human ctDNA in rat plasma derived from human glioblastoma multiforme stem-like cells in the rat brain and human hepatocellular carcinoma in the rat flank were found to have a shorter principal fragment length than the background rat cell-free DNA (134-144 bp vs. 167 bp, respectively). Subsequently, a similar shift in the fragment length of ctDNA in humans with melanoma and lung cancer was identified compared to healthy controls. Comparison of fragment lengths from cell-free DNA between a melanoma patient and healthy controls found that the BRAF V600E mutant allele occurred more commonly at a shorter fragment length than the fragment length of the wild-type allele (132-145 bp vs. 165 bp, respectively). Moreover, size-selecting for shorter cell-free DNA fragment lengths substantially increased the EGFR T790M mutant allele frequency in human lung cancer. These findings provide compelling evidence that experimental or bioinformatic isolation of a specific subset of fragment lengths from cell-free DNA may improve detection of ctDNA.Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor recurrence, and evaluate response to therapy solely through a non-invasive blood draw. However, detecting ctDNA against the normally occurring background of cell-free DNA derived from healthy cells has proven challenging, particularly in non-metastatic solid tumors. In this study, distinct differences in fragment length size between ctDNAs and normal cell-free DNA are defined. Human ctDNA in rat plasma derived from human glioblastoma multiforme stem-like cells in the rat brain and human hepatocellular carcinoma in the rat flank were found to have a shorter principal fragment length than the background rat cell-free DNA (134-144 bp vs. 167 bp, respectively). Subsequently, a similar shift in the fragment length of ctDNA in humans with melanoma and lung cancer was identified compared to healthy controls. Comparison of fragment lengths from cell-free DNA between a melanoma patient and healthy controls found that the BRAF V600E mutant allele occurred more commonly at a shorter fragment length than the fragment length of the wild-type allele (132-145 bp vs. 165 bp, respectively). Moreover, size-selecting for shorter cell-free DNA fragment lengths substantially increased the EGFR T790M mutant allele frequency in human lung cancer. These findings provide compelling evidence that experimental or bioinformatic isolation of a specific subset of fragment lengths from cell-free DNA may improve detection of ctDNA. Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor recurrence, and evaluate response to therapy solely through a non-invasive blood draw. However, detecting ctDNA against the normally occurring background of cell-free DNA derived from healthy cells has proven challenging, particularly in non-metastatic solid tumors. In this study, distinct differences in fragment length size between ctDNAs and normal cell-free DNA are defined. Human ctDNA in rat plasma derived from human glioblastoma multiforme stem-like cells in the rat brain and human hepatocellular carcinoma in the rat flank were found to have a shorter principal fragment length than the background rat cell-free DNA (134-144 bp vs. 167 bp, respectively). Subsequently, a similar shift in the fragment length of ctDNA in humans with melanoma and lung cancer was identified compared to healthy controls. Comparison of fragment lengths from cell-free DNA between a melanoma patient and healthy controls found that the BRAF V600E mutant allele occurred more commonly at a shorter fragment length than the fragment length of the wild-type allele (132-145 bp vs. 165 bp, respectively). Moreover, size-selecting for shorter cell-free DNA fragment lengths substantially increased the EGFR T790M mutant allele frequency in human lung cancer. These findings provide compelling evidence that experimental or bioinformatic isolation of a specific subset of fragment lengths from cell-free DNA may improve detection of ctDNA.
Audience	Academic
Author	Hellwig, Sabine Rostomily, Robert C. Baker, Daniel N. Welker, Noah C. Shendure, Jay Kitzman, Jacob O. Gligorich, Keith M. Underhill, Hunter R. Bronner, Mary P. Daza, Riza
AuthorAffiliation	1 Department of Pediatrics, Division of Medical Genetics, University of Utah, Salt Lake City, Utah, United States of America 6 ARUP Laboratories, Salt Lake City, Utah, United States of America 4 Department of Genome Sciences, University of Washington, Seattle, Washington, United States of America 3 Department of Neurological Surgery, University of Washington, Seattle, Washington, United States of America Brigham and Women's Hospital, UNITED STATES 2 Department of Radiology, University of Utah, Salt Lake City, Utah, United States of America 5 Department of Human Genetics, University of Michigan, Ann Arbor, Michigan, United States of America 7 Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America
AuthorAffiliation_xml	– name: 5 Department of Human Genetics, University of Michigan, Ann Arbor, Michigan, United States of America – name: Brigham and Women's Hospital, UNITED STATES – name: 1 Department of Pediatrics, Division of Medical Genetics, University of Utah, Salt Lake City, Utah, United States of America – name: 4 Department of Genome Sciences, University of Washington, Seattle, Washington, United States of America – name: 6 ARUP Laboratories, Salt Lake City, Utah, United States of America – name: 7 Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America – name: 2 Department of Radiology, University of Utah, Salt Lake City, Utah, United States of America – name: 3 Department of Neurological Surgery, University of Washington, Seattle, Washington, United States of America
Author_xml	– sequence: 1 givenname: Hunter R. surname: Underhill fullname: Underhill, Hunter R. – sequence: 2 givenname: Jacob O. surname: Kitzman fullname: Kitzman, Jacob O. – sequence: 3 givenname: Sabine surname: Hellwig fullname: Hellwig, Sabine – sequence: 4 givenname: Noah C. surname: Welker fullname: Welker, Noah C. – sequence: 5 givenname: Riza surname: Daza fullname: Daza, Riza – sequence: 6 givenname: Daniel N. orcidid: 0000-0002-0513-6893 surname: Baker fullname: Baker, Daniel N. – sequence: 7 givenname: Keith M. surname: Gligorich fullname: Gligorich, Keith M. – sequence: 8 givenname: Robert C. surname: Rostomily fullname: Rostomily, Robert C. – sequence: 9 givenname: Mary P. orcidid: 0000-0001-5574-4072 surname: Bronner fullname: Bronner, Mary P. – sequence: 10 givenname: Jay surname: Shendure fullname: Shendure, Jay
BackLink	https://www.ncbi.nlm.nih.gov/pubmed/27428049$$D View this record in MEDLINE/PubMed
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Snippet	Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor... Malignant tumors shed DNA into the circulation. The transient half-life of circulating tumor DNA (ctDNA) may afford the opportunity to diagnose, monitor...
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SubjectTerms	Alleles Animals Biology and life sciences Biomarkers, Tumor - blood Carcinoma, Hepatocellular - genetics Carcinoma, Hepatocellular - metabolism Cell Line, Tumor Deoxyribonucleic acid DNA DNA sequencing DNA, Neoplasm - blood DNA, Neoplasm - genetics Experiments Genes Genetic aspects Glioblastoma - blood Glioblastoma - genetics Hep G2 Cells Humans Liver Neoplasms - genetics Liver Neoplasms - metabolism Lung cancer Lung Neoplasms - genetics Lung Neoplasms - metabolism Magnetic Resonance Imaging Male Medicine and Health Sciences Melanoma Melanoma - genetics Melanoma - metabolism Metastasis Methods Mutation Neoplasm Transplantation Proto-Oncogene Proteins B-raf - genetics Rats Research and Analysis Methods Studies Tumors
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Title	Fragment Length of Circulating Tumor DNA
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