Absolute Quantitation of Met Using Mass Spectrometry for Clinical Application: Assay Precision, Stability, and Correlation with MET Gene Amplification in FFPE Tumor Tissue

被引:48
作者
Catenacci, Daniel V. T. [1 ]
Liao, Wei-Li [2 ]
Thyparambil, Sheeno [2 ]
Henderson, Les [1 ]
Xu, Peng [1 ]
Zhao, Lei [3 ]
Rambo, Brittany [1 ]
Hart, John [3 ]
Xiao, Shu-Yuan [3 ]
Bengali, Kathleen [2 ]
Uzzell, Jamar [2 ]
Darfler, Marlene [2 ]
Krizman, David B. [2 ]
Cecchi, Fabiola [4 ]
Bottaro, Donald P. [4 ]
Karrison, Theodore [5 ]
Veenstra, Timothy D. [2 ]
Hembrough, Todd [2 ]
Burrows, Jon [2 ]
机构
[1] Univ Chicago, Dept Med, Sect Hematol & Oncol, Chicago, IL 60637 USA
[2] OncoPlex Diagnost Inc, Rockville, MD USA
[3] Univ Chicago, Dept Pathol, Chicago, IL 60637 USA
[4] NCI, Urol Oncol Branch, NIH, Bethesda, MD 20892 USA
[5] Univ Chicago, Dept Hlth Sci, Chicago, IL 60637 USA
关键词
HEPATOCYTE GROWTH-FACTOR; STORED PARAFFIN SLIDES; CELL LUNG-CANCER; C-MET; GASTRIC-CANCER; SIGNALING PATHWAY; KINASE INHIBITOR; THERAPEUTIC TARGET; PROTEIN EXPRESSION; ADENOCARCINOMA;
D O I
10.1371/journal.pone.0100586
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: Overexpression of Met tyrosine kinase receptor is associated with poor prognosis. Overexpression, and particularly MET amplification, are predictive of response to Met-specific therapy in preclinical models. Immunohistochemistry (IHC) of formalin-fixed paraffin-embedded (FFPE) tissues is currently used to select for 'high Met' expressing tumors for Met inhibitor trials. IHC suffers from antibody non-specificity, lack of quantitative resolution, and, when quantifying multiple proteins, inefficient use of scarce tissue. Methods: After describing the development of the Liquid-Tissue-Selected Reaction Monitoring-mass spectrometry (LT-SRM-MS) Met assay, we evaluated the expression level of Met in 130 FFPE gastroesophageal cancer (GEC) tissues. We assessed the correlation of SRM Met expression to IHC and mean MET gene copy number (GCN)/nucleus or MET/CEP7 ratio by fluorescence in situ hybridization (FISH). Results: Proteomic mapping of recombinant Met identified (418)TEFTTALQR(426) as the optimal SRM peptide. Limits of detection (LOD) and quantitation (LOQ) for this peptide were 150 and 200 amol/mu g tumor protein, respectively. The assay demonstrated excellent precision and temporal stability of measurements in serial sections analyzed one year apart. Expression levels of 130 GEC tissues ranged (<150 amol/mu g to 4669.5 amol/mu g. High correlation was observed between SRM Met expression and both MET GCN and MET/CEP7 ratio as determined by FISH (n = 30; R-2 = 0.898). IHC did not correlate well with SRM (n = 44; R-2 = 0.537) nor FISH GCN (n = 31; R-2 = 0.509). A Met SRM level of >= 1500 amol/mu g was 100% sensitive (95% CI 0.69-1) and 100% specific (95% CI 0.92-1) for MET amplification. Conclusions: The Met SRM assay measured the absolute Met levels in clinical tissues with high precision. Compared to IHC, SRM provided a quantitative and linear measurement of Met expression, reliably distinguishing between non-amplified and amplified MET tumors. These results demonstrate a novel clinical tool for efficient tumor expression profiling, potentially leading to better informed therapeutic decisions for patients with GEC.
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页数:14
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[1]   COMPARATIVE MAPPING OF HUMAN ALPHOID SEQUENCES IN GREAT APES USING FLUORESCENCE IN-SITU HYBRIDIZATION [J].
ARCHIDIACONO, N ;
ANTONACCI, R ;
MARZELLA, R ;
FINELLI, P ;
LONOCE, A ;
ROCCHI, M .
GENOMICS, 1995, 25 (02) :477-484
[2]   c-Met ectodomain shedding rate correlates with malignant potential [J].
Athauda, Gagani ;
Giubellino, Alessio ;
Coleman, Jonathan A. ;
Horak, Christine ;
Steeg, Patricia S. ;
Lee, Ming-Jung ;
Trepel, Jane ;
Wimberly, Jennifer ;
Sun, Jan ;
Coxon, Angela ;
Burgess, Teresa L. ;
Bottaro, Donald P. .
CLINICAL CANCER RESEARCH, 2006, 12 (14) :4154-4162
[3]  
Bart K, 2014, ROCHE HALTS STUDY EX
[4]   Durable Complete Response of Metastatic Gastric Cancer with Anti-Met Therapy Followed by Resistance at Recurrence [J].
Catenacci, Daniel V. T. ;
Henderson, Les ;
Xiao, Shu-Yuan ;
Patel, Premal ;
Yauch, Robert L. ;
Hegde, Priti ;
Zha, Jiping ;
Pandita, Ajay ;
Peterson, Amy ;
Salgia, Ravi .
CANCER DISCOVERY, 2011, 1 (07) :573-579
[5]   RON (MST1R) is a novel prognostic marker and therapeutic target for gastroesophageal adenocarcinoma [J].
Catenacci, Daniel V. T. ;
Cervantes, Gustavo ;
Yala, Soheil ;
Nelson, Erik A. ;
El-Hashani, Essam ;
Kanteti, Rajani ;
El Dinali, Mohamed ;
Hasina, Rifat ;
Braegelmann, Johannes ;
Seiwert, Tanguy ;
Sanicola, Michele ;
Henderson, Les ;
Grushko, Tatyana A. ;
Olopade, Olufunmilayo ;
Karrison, Theodore ;
Bang, Yung-Jue ;
Kim, Woo Ho ;
Tretiakova, Maria ;
Vokes, Everett ;
Frank, David A. ;
Kindler, Hedy L. ;
Huet, Heather ;
Salgia, Ravi .
CANCER BIOLOGY & THERAPY, 2011, 12 (01) :9-46
[6]  
Catenacci DVT XP, 2012, 24 EORTC NCI AACR S
[7]   Targeting the HGF/Met signaling pathway in cancer therapy [J].
Cecchi, Fabiola ;
Rabe, Danie C. ;
Bottaro, Donald P. .
EXPERT OPINION ON THERAPEUTIC TARGETS, 2012, 16 (06) :553-572
[8]   BMS-777607, a Small-Molecule Met Kinase Inhibitor, Suppresses Hepatocyte Growth Factor-Stimulated Prostate Cancer Metastatic Phenotype In vitro [J].
Dai, Yao ;
Siemann, Dietmar W. .
MOLECULAR CANCER THERAPEUTICS, 2010, 9 (06) :1554-1561
[9]   Discovery of a Novel Mode of Protein Kinase Inhibition Characterized by the Mechanism of Inhibition of Human Mesenchymal-epithelial Transition Factor (c-Met) Protein Autophosphorylation by ARQ 197 [J].
Eathiraj, Sudharshan ;
Palma, Rocio ;
Volckova, Erika ;
Hirschi, Marscha ;
France, Dennis S. ;
Ashwell, Mark A. ;
Chan, Thomas C. K. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2011, 286 (23) :20666-20676
[10]   Novel Therapeutic Inhibitors of the c-Met Signaling Pathway in Cancer [J].
Eder, Joseph Paul ;
Woude, George F. Vande ;
Boerner, Scott A. ;
LoRusso, Patricia M. .
CLINICAL CANCER RESEARCH, 2009, 15 (07) :2207-2214