title: Impact of protein stability, cellular localization, and abundance on proteomic detection of tumor-derived proteins in plasma.
abstract: Tumor-derived, circulating proteins are potentially useful as biomarkers for detection of cancer, for monitoring of disease progression, regression and recurrence, and for assessment of therapeutic response. Here we interrogated how a protein's stability, cellular localization, and abundance affect its observability in blood by mass-spectrometry-based proteomics techniques. We performed proteomic profiling on tumors and plasma from two different xenograft mouse models. A statistical analysis of this data revealed protein properties indicative of the detection level in plasma. Though 20% of the proteins identified in plasma were tumor-derived, only 5% of the proteins observed in the tumor tissue were found in plasma. Both intracellular and extracellular tumor proteins were observed in plasma; however, after normalizing for tumor abundance, extracellular proteins were seven times more likely to be detected. Although proteins that were more abundant in the tumor were also more likely to be observed in plasma, the relationship was nonlinear: Doubling the spectral count increased detection rate by only 50%. Many secreted proteins, even those with relatively low spectral count, were observed in plasma, but few low abundance intracellular proteins were observed. Proteins predicted to be stable by dipeptide composition were significantly more likely to be identified in plasma than less stable proteins. The number of tryptic peptides in a protein was not significantly related to the chance of a protein being observed in plasma. Quantitative comparison of large versus small tumors revealed that the abundance of proteins in plasma as measured by spectral count was associated with the tumor size, but the relationship was not one-to-one; a 3-fold decrease in tumor size resulted in a 16-fold decrease in protein abundance in plasma. This study provides quantitative support for a tumor-derived marker prioritization strategy that favors secreted and stable proteins over all but the most abundant intracellular proteins.
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.
authors.forename: Qiaojun
authors.initials: Q
authors.lastname: Fang
authors.forename: Kian
authors.initials: K
authors.lastname: Kani
authors.forename: Vitor M
authors.initials: VM
authors.lastname: Faca
authors.forename: Wenxuan
authors.initials: W
authors.lastname: Zhang
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhang
authors.forename: Anjali
authors.initials: A
authors.lastname: Jain
authors.forename: Sam
authors.initials: S
authors.lastname: Hanash
authors.forename: David B
authors.initials: DB
authors.lastname: Agus
authors.forename: Martin W
authors.initials: MW
authors.lastname: McIntosh
authors.forename: Parag
authors.initials: P
authors.lastname: Mallick
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use; D014408:Biomarkers, Tumor / Q000378:metabolism; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm; D005260:Female; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism*; D055550:Protein Stability; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D012878:Skin Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D001798:Blood Proteins; D009363:Neoplasm Proteins; D020543:Proteome