Combined blood/tissue analysis for cancer biomarker discovery: application to renal cell carcinoma.
A method that relies on subtractive tissue-directed shot-gun proteomics to identify tumor proteins in the blood of a patient newly diagnosed with cancer is described. To avoid analytical and statistical biases caused by physiologic variability of protein expression in the human population, this method was applied on clinical specimens obtained from a single patient diagnosed with nonmetastatic renal cell carcinoma (RCC). The proteomes extracted from tumor, normal adjacent tissue and preoperative plasma were analyzed using 2D-liquid chromatography-mass spectrometry (LC-MS). The lists of identified proteins were filtered to discover proteins that (i) were found in the tumor but not normal tissue, (ii) were identified in matching plasma, and (iii) whose spectral count was higher in tumor tissue than plasma. These filtering criteria resulted in identification of eight tumor proteins in the blood. Subsequent Western-blot analysis confirmed the presence of cadherin-5, cadherin-11, DEAD-box protein-23, and pyruvate kinase in the blood of the patient in the study as well as in the blood of four other patients diagnosed with RCC. These results demonstrate the utility of a combined blood/tissue analysis strategy that permits the detection of tumor proteins in the blood of a patient diagnosed with RCC.
Johann
Donald J
DJ
Wei
Bih-Rong
BR
Prieto
DaRue A
DA
Chan
King C
KC
Ye
Xiaying
X
Valera
Vladimir A
VA
Simpson
R Mark
RM
Rudnick
Paul A
PA
Xiao
Zhen
Z
Issaq
Haleem J
HJ
Linehan
W Marston
WM
Stein
Stephen E
SE
Veenstra
Timothy D
TD
Blonder
Josip
J
D014408:Biomarkers, Tumor / Q000032:analysis / Q000378:metabolism*; D002292:Carcinoma, Renal Cell / Q000097:blood* / Q000175:diagnosis; D002853:Chromatography, Liquid; D006801:Humans; D007680:Kidney Neoplasms / Q000097:blood* / Q000175:diagnosis; D013058:Mass Spectrometry
D014408:Biomarkers, Tumor