Phosphoproteins in extracellular vesicles as candidate markers for breast cancer
The state of protein phosphorylation can be a key determinant of cellular physiology such as early-stage cancer, but the development of phosphoproteins in biofluids for disease diagnosis remains elusive. Here we demonstrate a strategy to isolate and identify phosphoproteins in extracellular vesicles...
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| Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 114; no. 12; pp. 3175 - 3180 |
|---|---|
| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
National Academy of Sciences
21.03.2017
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| Subjects | |
| Online Access | Get full text |
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| Abstract | The state of protein phosphorylation can be a key determinant of cellular physiology such as early-stage cancer, but the development of phosphoproteins in biofluids for disease diagnosis remains elusive. Here we demonstrate a strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs isolated from small volumes of plasma samples. Using label-free quantitative phosphoproteomics, we identified 144 phosphoproteins in plasma EVs that are significantly higher in patients diagnosed with breast cancer compared with healthy controls. Several biomarkers were validated in individual patients using paralleled reaction monitoring for targeted quantitation. This study demonstrates that the development of phosphoproteins in plasma EV as disease biomarkers is highly feasible and may transform cancer screening and monitoring. |
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| AbstractList | Protein phosphorylation is a major regulatory mechanism for many cellular functions, but no phosphoprotein in biofluids has been developed for disease diagnosis because of the presence of active phosphatases. This study presents a general strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs from small volumes of plasma samples and more than 100 phosphoproteins in plasma EVs that are significantly higher in patients diagnosed with breast cancer as compared with healthy controls. This study demonstrates that the development of phosphoproteins in plasma EVs as disease biomarkers is highly feasible and may transform cancer screening and monitoring.
The state of protein phosphorylation can be a key determinant of cellular physiology such as early-stage cancer, but the development of phosphoproteins in biofluids for disease diagnosis remains elusive. Here we demonstrate a strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs isolated from small volumes of plasma samples. Using label-free quantitative phosphoproteomics, we identified 144 phosphoproteins in plasma EVs that are significantly higher in patients diagnosed with breast cancer compared with healthy controls. Several biomarkers were validated in individual patients using paralleled reaction monitoring for targeted quantitation. This study demonstrates that the development of phosphoproteins in plasma EV as disease biomarkers is highly feasible and may transform cancer screening and monitoring. The state of protein phosphorylation can be a key determinant of cellular physiology such as early-stage cancer, but the development of phosphoproteins in biofluids for disease diagnosis remains elusive. Here we demonstrate a strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs isolated from small volumes of plasma samples. Using label-free quantitative phosphoproteomics, we identified 144 phosphoproteins in plasma EVs that are significantly higher in patients diagnosed with breast cancer compared with healthy controls. Several biomarkers were validated in individual patients using paralleled reaction monitoring for targeted quantitation. This study demonstrates that the development of phosphoproteins in plasma EV as disease biomarkers is highly feasible and may transform cancer screening and monitoring. The state of protein phosphorylation can be a key determinant of cellular physiology such as early-stage cancer, but the development of phosphoproteins in biofluids for disease diagnosis remains elusive. Here we demonstrate a strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs isolated from small volumes of plasma samples. Using label-free quantitative phosphoproteomics, we identified 144 phosphoproteins in plasma EVs that are significantly higher in patients diagnosed with breast cancer compared with healthy controls. Several biomarkers were validated in individual patients using paralleled reaction monitoring for targeted quantitation. This study demonstrates that the development of phosphoproteins in plasma EV as disease biomarkers is highly feasible and may transform cancer screening and monitoring.The state of protein phosphorylation can be a key determinant of cellular physiology such as early-stage cancer, but the development of phosphoproteins in biofluids for disease diagnosis remains elusive. Here we demonstrate a strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs isolated from small volumes of plasma samples. Using label-free quantitative phosphoproteomics, we identified 144 phosphoproteins in plasma EVs that are significantly higher in patients diagnosed with breast cancer compared with healthy controls. Several biomarkers were validated in individual patients using paralleled reaction monitoring for targeted quantitation. This study demonstrates that the development of phosphoproteins in plasma EV as disease biomarkers is highly feasible and may transform cancer screening and monitoring. |
| Author | Hsu, Chuan-Chih Chen, I-Hsuan Tao, W. Andy Pan, Li Zhu, Jian-Kang Andaluz, Hillary Xue, Liang Iliuk, Anton B. Wendt, Michael K. Paez, Juan Sebastian Paez |
| Author_xml | – sequence: 1 givenname: I-Hsuan surname: Chen fullname: Chen, I-Hsuan organization: Department of Biochemistry, Purdue University, West Lafayette, IN 47907 – sequence: 2 givenname: Liang surname: Xue fullname: Xue, Liang organization: Department of Biochemistry, Purdue University, West Lafayette, IN 47907 – sequence: 3 givenname: Chuan-Chih surname: Hsu fullname: Hsu, Chuan-Chih organization: Department of Biochemistry, Purdue University, West Lafayette, IN 47907 – sequence: 4 givenname: Juan Sebastian Paez surname: Paez fullname: Paez, Juan Sebastian Paez organization: Department of Biochemistry, Purdue University, West Lafayette, IN 47907 – sequence: 5 givenname: Li surname: Pan fullname: Pan, Li organization: Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN 47907 – sequence: 6 givenname: Hillary surname: Andaluz fullname: Andaluz, Hillary organization: Department of Chemistry, Purdue University, West Lafayette, IN 47907 – sequence: 7 givenname: Michael K. surname: Wendt fullname: Wendt, Michael K. organization: Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN 47907 – sequence: 8 givenname: Anton B. surname: Iliuk fullname: Iliuk, Anton B. organization: Department of Innovations, Tymora Analytical Operations, West Lafayette, IN 47906 – sequence: 9 givenname: Jian-Kang surname: Zhu fullname: Zhu, Jian-Kang organization: Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai 201602, China – sequence: 10 givenname: W. Andy surname: Tao fullname: Tao, W. Andy organization: Purdue Center for Cancer Research, Purdue University, West Lafayette, IN 47907 |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/28270605$$D View this record in MEDLINE/PubMed |
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| Copyright | Volumes 1–89 and 106–114, copyright as a collective work only; author(s) retains copyright to individual articles Copyright National Academy of Sciences Mar 21, 2017 |
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| Keywords | phosphoproteins mass spectrometry extracellular vesicles proteomics biomarker |
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| Notes | SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 14 ObjectType-Article-1 ObjectType-Feature-2 content type line 23 Contributed by Jian-Kang Zhu, February 1, 2017 (sent for review November 1, 2016; reviewed by Natalie G. Ahn, Bernd Bodenmiller, and Jim Bruce) Author contributions: I.-H.C., L.P., A.B.I., J.-K.Z., and W.A.T. designed research; I.-H.C., L.X., C.-C.H., H.A., and A.B.I. performed research; I.-H.C., J.S.P.P., M.K.W., J.-K.Z., and W.A.T. analyzed data; and I.-H.C. and W.A.T. wrote the paper. Reviewers: N.G.A., University of Colorado; B.B., University of Zurich; and J.B., University of Washington. |
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| Title | Phosphoproteins in extracellular vesicles as candidate markers for breast cancer |
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