title: Phosphoproteins in extracellular vesicles as candidate markers for breast cancer.
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.
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN 47907.
authors.forename: I-Hsuan
authors.initials: IH
authors.lastname: Chen
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN 47907.
authors.forename: Liang
authors.initials: L
authors.lastname: Xue
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN 47907.
authors.forename: Chuan-Chih
authors.initials: CC
authors.lastname: Hsu
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN 47907.
authors.forename: Juan Sebastian Paez
authors.initials: JS
authors.lastname: Paez
authors.affiliation: Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN 47907.
authors.forename: Li
authors.initials: L
authors.lastname: Pan
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, IN 47907.
authors.forename: Hillary
authors.initials: H
authors.lastname: Andaluz
authors.affiliation: Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN 47907.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Wendt
authors.affiliation: Department of Innovations, Tymora Analytical Operations, West Lafayette, IN 47906.
authors.forename: Anton B
authors.initials: AB
authors.lastname: Iliuk
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN 47907; jkzhu@purdue.edu watao@purdue.edu.|Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907.|Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai 201602, China.
authors.forename: Jian-Kang
authors.identifier: 0000-0001-5134-731X
authors.initials: JK
authors.lastname: Zhu
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, IN 47907; jkzhu@purdue.edu watao@purdue.edu.|Department of Medicinal Chemistry & Molecular Pharmacology, Purdue University, West Lafayette, IN 47907.|Department of Chemistry, Purdue University, West Lafayette, IN 47907.|Purdue Center for Cancer Research, Purdue University, West Lafayette, IN 47907.
authors.forename: W Andy
authors.initials: WA
authors.lastname: Tao
mesh_terms: D015415:Biomarkers; D014408:Biomarkers, Tumor; D001798:Blood Proteins; D001943:Breast Neoplasms / Q000097:blood / Q000378:metabolism*; D016022:Case-Control Studies; D016000:Cluster Analysis; D019295:Computational Biology / Q000379:methods; D055354:Exosomes / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000378:metabolism*; D005260:Female; D006801:Humans; D010748:Phosphopeptides / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry; D057188:Workflow
chemical_list: D015415:Biomarkers; D014408:Biomarkers, Tumor; D001798:Blood Proteins; D010748:Phosphopeptides; D010750:Phosphoproteins; D020543:Proteome
keywords: biomarker; extracellular vesicles; mass spectrometry; phosphoproteins; proteomics