Upload
marvin-holland
View
215
Download
0
Embed Size (px)
Citation preview
Pfetin, as a Prognostic Biomarker of Pfetin, as a Prognostic Biomarker of Gastrointestinal Stromal Tumors Gastrointestinal Stromal Tumors RevRev
ealedealed by Proteomics by Proteomics
Yoshiyuki SueharaYoshiyuki Suehara1 3 41 3 4, Kunihiko Seki, Kunihiko Seki22, Kiyonaga Fujii, Kiyonaga Fujii11, Tadashi Hase, Tadashi Hasegawagawa2 2 , Tadakazu Shimoda, Tadakazu Shimoda22, Yasuo Beppu, Yasuo Beppu44, Akira Kawai, Akira Kawai44, Setsuo Hir, Setsuo Hirohashi ohashi 11,, Tadashi KondoTadashi Kondo11
1. Proteome Bioinformatics Project, National Cancer Center Research Institute, Tokyo, Japan2. Clinical Laboratory Division, National Cancer Center Research Hospital, Tokyo, Japan3. Department of Orthopedic Surgery, Juntendo University School of Medicine, Tokyo, Japan4. Orthopedic Surgery Division, National Cancer Center Hospital, Tokyo, Japan
MWMW(KDa)(KDa)
Isoelectric focuIsoelectric focusingsing
8080
5656
3636
3030
1818
pH 4.0pH 4.0 pH 7.0pH 7.0
Procedure of Proteomics StudyProcedure of Proteomics Study
1500-2000 1500-2000 spotsspots
Protein was extracted and labeled with CyDye
2D-Image
Data-MiningProtein Protein spots spots were were identifidentifiedied
High-speed/-throughput 1D-RP mLC/NSI-MS/MS Ana
lysis
Mass Spectrometry
Surgical Specimen
Back Ground (GISTs)Back Ground (GISTs)• GISTs are the most common primary mesenchymal tumor o
f the digestive tract.• It is genetically characterized by the presence of mutations a
nd overexpression of KIT and, clinically, characterized by their significant response to treatment with imatinib.
• However, there are certain populations of patients who can be treated only by simple resection and do not need imatinib treatment.
The aim of this study is to develop prognostic biomarkers for GISTs, by means of proteomic approach, and identify high-risk patients who would need further adjuvant treatment such as imatinib.
StrategyStrategy“Poor prognosis GISTs”
= Metastasis within 1 year
(8 samples)
“Good prognosis GISTs”
= No-metastasis over 2 years
Low or IM Risk group *
(9 samples)
* Pathological factors (Risk Classification-> Hasegawa T. et al: Human Pathology. 2002)
vs
Identify the informative Identify the informative spotsspots
Confirmed these resultsConfirmed these resultsWestern-blot, Immunochemical Western-blot, Immunochemical
studystudy
Large-scale sample set (Immunohistochemical study)Large-scale sample set (Immunohistochemical study)Verified the power of biomarkerVerified the power of biomarker
Cluster Analysis 43 spots Cluster Analysis 43 spots (p < 0.01)(p < 0.01)
Poor-prognosis GISTs
Good-prognosis GISTs PfetinPfetin
1513 spots
43 spotsWilcoxon test
Poor-prognosis GISTsMetastasis within 1 year(samples 1-8)
Good-prognosis GISTs No-metastasis over 2 years Low or IM Risk group(samples 9-17)
Immunohistochemical studyImmunohistochemical study(Pfetin antibody)
Western Western BlottingBlotting
Poor prognosis GISTs Good prognosis GISTs
(Pfetin antibody)
Good prognosis GISTs
Poor prognosis GISTs
210 cases, M0, primary samples
Metastasis-Free Survival of the M0 Metastasis-Free Survival of the M0 GISTsGISTs Patients Patients According to the Expression of Pfetin According to the Expression of Pfetin (NCC :210 cases)(NCC :210 cases)
Suehara Y et al. Clin Cancer Res (2008) 14: 1707-1717Nature Clinical Practice Oncology (2008) 5, 364-365
5-year: 93.9%
5-year: 36.2%
P < 0.0001
(Immunohistochemical Study)(Immunohistochemical Study)
Overall Survival of the M0 GISTs Patients Overall Survival of the M0 GISTs Patients According to the Expression of Pfetin According to the Expression of Pfetin (NCC :210 cases)(NCC :210 cases)
Suehara Y et al. Clin Cancer Res (2008) 14: 1707-1717Nature Clinical Practice Oncology (2008) 5, 364-365
P < 0.0001
5-year: 97.2%
5-year: 76.5%
210 cases, M0, primary samples
(Immunohistochemical Study)(Immunohistochemical Study)
Metastasis-Free Survival Curve of GISTsMetastasis-Free Survival Curve of GISTs According to Risk Classification (NCC :210 case According to Risk Classification (NCC :210 case
s)s)
N=110 N=46
N=54
High risk
Suehara Y et al. Clin Cancer Res (2008) 14: 1707-1717Nature Clinical Practice Oncology (2008) 5, 364-365
Intermediate riskLow risk
Metastasis-Free Survival Curve of GISTsMetastasis-Free Survival Curve of GISTs According to Pfetin Immunochemical Stain (NCC :210 case According to Pfetin Immunochemical Stain (NCC :210 case
s)s)
High risk
N=110 N=46
N=54
Pfetin positive (N=100)
Pfetin positive (N=27)
Pfetin positive (N=44)
Pfetin negative (N=10) Pfetin negative (N=2)
Pfetin negative (N=27)
P = 0.0002
Suehara Y et al. Clin Cancer Res (2008) 14: 1707-1717Nature Clinical Practice Oncology (2008) 5, 364-365
Intermediate riskLow risk
P<0.0001 P=0.0109
P=0.0002
Multivariate Analysis of Multivariate Analysis of Metastasis Free Survival by Metastasis Free Survival by
Cox RegressionCox RegressionMultivariate analysis of metastasis free survivl by Cox regression
Variable P value Relative risk 95% confidence interval
SiteStomach
Non-stomach 0.0270 2.21 1.09-4.49Size
<5 cm5-10cm15cm< 0.0070 2.05 1.22-3.44
DifferentationScore 1Score 2 <0.0001 10.40 3.68-29.45
PfetinPositive
Negative 0.0020 3.75 1.60-8.81
ConclusiConclusionon
• We examined the proteomic profile of GISTs accWe examined the proteomic profile of GISTs according to prognosis using both 2D-DIGE and clording to prognosis using both 2D-DIGE and clinical data.inical data.
• Pfetin expression was a significant predictor Pfetin expression was a significant predictor for metastasis and survival of patients with Gfor metastasis and survival of patients with GISTs.ISTs.
• Pfetin can be a strong candidate for a biomarkPfetin can be a strong candidate for a biomarker to predict the metastasis in GISTs patienter to predict the metastasis in GISTs patients. s.