Identification of a Predictive Biomarker for Hematologic Toxicities of Gemcitabine
JOURNAL OF CLINICAL ONCOLOGY
2009; 27 (13): 2261-2268
Identification of Adipophilin as a Potential Plasma Biomarker for Colorectal Cancer Using Label-Free Quantitative Mass Spectrometry and Protein Microarray
CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION
2011; 20 (10): 2195-2203
Gemcitabine monotherapy is the current standard for patients with advanced pancreatic cancer, but the occurrence of severe neutropenia and thrombocytopenia can sometimes be life threatening. This study aimed to discover a new diagnostic method for predicting the hematologic toxicities of gemcitabine.Using quantitative mass spectrometry (MS), we compared the baseline plasma proteomes of 25 patients who had developed severe hematologic adverse events (grade 3 to 4 neutropenia and/or grade 2 to 4 thrombocytopenia) within the first two cycles of gemcitabine with those of 22 patients who had not (grade 0).We identified 757 peptide peaks whose intensities were significantly different (P < .001, Welch t test) among a total of 60,888. The MS peak with the highest statistical significance (P = .0000282) was revealed to be derived from haptoglobin by tandem MS. A scoring system (nomogram) based on the values of haptoglobin, haptoglobin phenotype, neutrophil count, platelet count, and body-surface area was constructed to estimate the risk of hematologic adverse events (grade 3 to 4 neutropenia and/or grade 2 to 4 thrombocytopenia) with an area under curve value of 0.782 in a cohort of 166 patients with pancreatic cancer. Predictive ability of the system was confirmed in two independent validation cohorts consisting of 87 and 52 patients with area under the curve values of 0.655 and 0.747, respectively.Although the precise mechanism responsible for the correlation of haptoglobin with the future onset of hematologic toxicities remains to be clarified, our prediction model seems to have high practical utility for tailoring the treatment of patients receiving gemcitabine.
View details for DOI 10.1200/JCO.2008.19.9745
View details for Web of Science ID 000266195000024
View details for PubMedID 19289617
Reduced Plasma Level of CXC Chemokine Ligand 7 in Patients with Pancreatic Cancer
CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION
2011; 20 (1): 160-171
The aim of this study was to identify a new plasma biomarker for use in early detection of colorectal cancer.Using the combination of hollow fiber membrane (HFM)-based low-molecular weight protein enrichment and two-dimensional image converted analysis of liquid chromatography and mass spectrometry (2DICAL), we compared the plasma proteome of 22 colorectal cancer patients with those of 21 healthy controls. An identified biomarker candidate was then validated in two larger cohorts [validation-1 (n = 210) and validation-2 (n = 113)] using a high-density reverse-phase protein microarray.From a total of 53,009 mass peaks, we identified 103 with an area under curve (AUC) value of 0.80 or higher that could distinguish cancer patients from healthy controls. A peak that increased in colorectal cancer patients, with an AUC of 0.81 and P value of 0.0004 (Mann-Whitney U test), was identified as a product of the PLIN2 gene [also known as perilipin-2, adipose differentiation-related protein (ADRP), or adipophilin]. An increase in plasma adipophilin was consistently observed in colorectal cancer patients, including those with stage I or stage II disease (P < 0.0001, Welch's t test). Immunohistochemical analysis revealed that adipophilin is expressed primarily in the basal sides of colorectal cancer cells forming polarized tubular structures, and that it is absent from adjacent normal intestinal mucosae.Adipophilin is a plasma biomarker potentially useful for the detection of early-stage colorectal cancer.The combination of HFM and 2DICAL enables the comprehensive analysis of plasma proteins and is ideal for use in all biomarker discovery studies.
View details for DOI 10.1158/1055-9965.EPI-11-0400
View details for Web of Science ID 000295717900025
View details for PubMedID 21828233
Phase II Study of Bolus 5-Fluorouracil and Leucovorin Combined with Weekly Paclitaxel as First-Line Therapy for Advanced Gastric Cancer
2011; 81 (5-6): 291-297
Early detection is essential to improve the outcome of patients with pancreatic cancer. A noninvasive and cost-effective diagnostic test using plasma/serum biomarkers would facilitate the detection of pancreatic cancer at the early stage.Using a novel combination of hollow fiber membrane-based low-molecular-weight protein enrichment and LC-MS-based quantitative shotgun proteomics, we compared the plasma proteome between 24 patients with pancreatic cancer and 21 healthy controls (training cohort). An identified biomarker candidate was then subjected to a large blinded independent validation (n = 237, validation cohort) using a high-density reverse-phase protein microarray.Among a total of 53,009 MS peaks, we identified a peptide derived from CXC chemokine ligand 7 (CXCL7) that was significantly reduced in pancreatic cancer patients, showing an area under curve (AUC) value of 0.84 and a P value of 0.00005 (Mann-Whitney U test). Reduction of the CXCL7 protein was consistently observed in pancreatic cancer patients including those with stage I and II disease in the validation cohort (P < 0.0001). The plasma level of CXCL7 was independent from that of CA19-9 (Pearson's r = 0.289), and combination with CXCL7 significantly improved the AUC value of CA19-9 to 0.961 (P = 0.002).We identified a significant decrease of the plasma CXCL7 level in patients with pancreatic cancer, and combination of CA19-9 with CXCL7 improved the discriminatory power of the former for pancreatic cancer.The present findings may provide a new diagnostic option for pancreatic cancer and facilitate early detection of the disease.
View details for DOI 10.1158/1055-9965.EPI-10-0397
View details for Web of Science ID 000285972800017
View details for PubMedID 21148121
[Impact of HER2, EGFR, IGF-1R, and VEGFR expressions on the outcome of chemotherapy for advanced gastric cancer].
Gan to kagaku ryoho. Cancer & chemotherapy
2010; 37 (8): 1489-1496
We evaluated the efficacy and safety of bolus 5-fluorouracil (5-FU) and leucovorin combined with weekly paclitaxel (FLTAX) in advanced gastric cancer (GC) patients.Patients with untreated stage IV GC received paclitaxel 80 mg/m(2) as a 1-hour infusion, followed by 5-FU 600 mg/m(2) as a bolus infusion and L-leucovorin 250 mg/m(2) as a 2-hour infusion on days 1, 8 and 15. Treatment cycles were repeated every 28 days. The primary endpoint was response rate.Thirty-five patients were enrolled. The median age was 62 years (range 34-75). Twenty-one patients (60%) had diffuse-type cancer and 11 had peritoneal metastasis. The confirmed response rate was 43% (95% CI 26-61) with 15 partial responses. Stable disease was observed in 16 (46%) patients. Median progression-free survival and overall survival were 6.8 months (95% CI 5.8-7.4) and 16.2 months (95% CI 10.0-22.8), respectively. Grade 3-4 adverse events were: neutropenia (54%), febrile neutropenia (3%), diarrhea (6%) and sensory neuropathy (11%).FLTAX showed a desirable safety profile, and the efficacy against advanced GC was encouraging. FLTAX may be a good option for GC patients with deteriorated general condition, and a randomized clinical trial in such patients is currently underway.
View details for DOI 10.1159/000334462
View details for Web of Science ID 000300098300002
View details for PubMedID 22134040
Survival Prediction for Pancreatic Cancer Patients Receiving Gemcitabine Treatment
MOLECULAR & CELLULAR PROTEOMICS
2010; 9 (4): 695-704
We aimed to elucidate the clinical significance of the expressions of HER2, epidermal growth factor receptor (EGFR), insulin-like growth factor receptor (IGF-1R), and vascular endothelial growth factor receptor 1, 2, 3 (VEGF-R1, VEGF-R2, and VEGF-R3) in gastric cancer. The study group comprised 57 patients who had undergone gastrectomy at the National Cancer Center Hospital and subsequently received first-line chemotherapy (S-1 monotherapy [n=29] or irinotecan+cisplatin [n=28]) for recurrent or residual tumors. We performed immunohistochemical analysis of formalin-fixed paraffinembedded specimens of surgically removed primary tumors to determine the expressions of HER2, EGFR, IGF-1R, and VEGFR1 in tumor cells and the expressions of VEGF-R1, VEGF-R2, and VEGF-R3 in tumor stromal vessels. The expressions of HER2 (p=0.017) and IGF-1R (p=0.025) were significantly more common in intestinal type tumors than in diffuse type. The protein expressions did not correlate with tumor response in either chemotherapy-regimen group. Among the patients who underwent S-1 monotherapy, those with cytoplasmic VEGF-R1-positive tumors had significantly shorter progression-free survival (logrank, p=0.017). In the survival analysis of all the patients, coexpression of membranous IGF-1R and VEGF-R3 in stromal vessels was the most significant predictor of poor survival (hazard ratio, 1.82; 95% CI, 1.31-2.63; p<0.001). The results of our study will facilitate more efficient use of molecular targeted agents in patients with gastric cancer.
View details for PubMedID 20716873
Prolyl 4-Hydroxylation of alpha-Fibrinogen A NOVEL PROTEIN MODIFICATION REVEALED BY PLASMA PROTEOMICS
JOURNAL OF BIOLOGICAL CHEMISTRY
2009; 284 (42): 29041-29049
Although gemcitabine monotherapy is the standard treatment for advanced pancreatic cancer, patient outcome varies significantly, and a considerable number do not benefit adequately. We therefore searched for new biomarkers predictive of overall patient survival. Using LC-MS, we compared the base-line plasma proteome between 29 representative patients with advanced pancreatic cancer who died within 100 days and 31 patients who survived for more than 400 days after receiving at least two cycles of the same gemcitabine monotherapy. Identified biomarker candidates were then challenged in a larger cohort of 304 patients treated with the same protocol using reverse-phase protein microarray. Among a total of 45,277 peptide peaks, we identified 637 peaks whose intensities differed significantly between the two groups (p < 0.001, Welch's t test). Two MS peaks with the highest statistical significance (p = 2.6 x 10(-4) and p = 5.0 x 10(-4)) were revealed to be derived from alpha(1)-antitrypsin and alpha(1)-antichymotrypsin, respectively. The levels of alpha(1)-antitrypsin (p = 8.9 x 10(-8)) and alpha(1)-antichymotrypsin (p = 0.001) were significantly correlated with the overall survival of the 304 patients. We selected alpha(1)-antitrypsin (p = 0.0001), leukocyte count (p = 0.066), alkaline phosphatase (p = 8.3 x 10(-8)), and performance status (p = 0.003) using multivariate Cox regression analysis and constructed a scoring system (nomogram) that was able to identify a group of high risk patients having a short median survival time of 150 days (95% confidence interval, 123-187 days; p = 2.0 x 10(-15), log rank test). The accuracy of this model for prognostication was internally validated and showed good calibration and discrimination with a bootstrap-corrected concordance index of 0.672. In conclusion, an increased level of alpha(1)-antitrypsin is a biomarker that predicts short overall survival of patients with advanced pancreatic cancer receiving gemcitabine monotherapy. Although an external validation study will be necessary, the current model may be useful for identifying patients unsuitable for the standardized therapy.
View details for DOI 10.1074/mcp.M900234-MCP200
View details for Web of Science ID 000276379400009
View details for PubMedID 20061307
A phase I study of bolus 5-fluorouracil and leucovorin combined with weekly paclitaxel (FLTAX) as first-line therapy for advanced gastric cancer
JAPANESE JOURNAL OF CLINICAL ONCOLOGY
2008; 38 (8): 540-546
Plasma proteome analysis requires sufficient power to compare numerous samples and detect changes in protein modification, because the protein content of human samples varies significantly among individuals, and many plasma proteins undergo changes in the bloodstream. A label-free proteomics platform developed in our laboratory, termed "Two-Dimensional Image Converted Analysis of Liquid chromatography and mass spectrometry (2DICAL)," is capable of these tasks. Here, we describe successful detection of novel prolyl hydroxylation of alpha-fibrinogen using 2DICAL, based on comparison of plasma samples of 38 pancreatic cancer patients and 39 healthy subjects. Using a newly generated monoclonal antibody 11A5, we confirmed the increase in prolyl-hydroxylated alpha-fibrinogen plasma levels and identified prolyl 4-hydroxylase A1 as a key enzyme for the modification. Competitive enzyme-linked immunosorbent assay of 685 blood samples revealed dynamic changes in prolyl-hydroxylated alpha-fibrinogen plasma level depending on clinical status. Prolyl-hydroxylated alpha-fibrinogen is presumably controlled by multiple biological mechanisms, which remain to be clarified in future studies.
View details for DOI 10.1074/jbc.M109.041749
View details for Web of Science ID 000270678900061
View details for PubMedID 19696023
Impact of insulin-like growth factor type 1 receptor, epidermal growth factor receptor, and HER2 expressions on outcomes of patients with gastric cancer
CLINICAL CANCER RESEARCH
2008; 14 (10): 3022-3029
To determine the dose-limiting toxicity (DLT) and the maximum-tolerated dose (MTD) of combination chemotherapy with leucovorin-modulated weekly bolus 5-fluorouracil (5-FU) and weekly paclitaxel in patients with advanced gastric cancer (GC).Chemotherapy-naive patients with histologically proven metastatic or recurrent GC were enrolled. Paclitaxel was administered as a 1-h intravenous (i.v.) infusion followed by 5-FU as a bolus i.v. infusion on Days 1, 8 and 15. A 2-h i.v. infusion of l-leucovorin was started at the same time as the paclitaxel infusion on Days 1, 8 and 15. Treatment cycles were repeated every 28 days until disease progression or unacceptable toxicity occurred. Patients were scheduled to receive 5-FU, l-leucovorin and paclitaxel at four dose levels (mg/m(2)/week): 500/250/60 (level 1), 500/250/80 (level 2), 600/250/80 (level 3) and 600/250/100 (level 4), respectively.Eighteen patients were enrolled. During the first cycle of the highest dose level (level 4), two of the six patients had DLT involving Grade 3 diarrhea and Grade 3 skin rash. Furthermore, three of the four patients who received the second consecutive cycle of treatment at dose level 4 had Grade 4 neutropenia. Dose level 3 was thus determined to be the MTD. Eleven (61%) of the 18 patients had partial responses, and the median progression-free survival time was 6.8 months.The MTD and the recommended dose for phase II studies of this regimen were determined to be 5-FU 600 mg/m(2)/week, l-leucovorin 250 mg/m(2)/week and paclitaxel 80 mg/m(2)/week.
View details for DOI 10.1093/jjco/hyn062
View details for Web of Science ID 000258864100006
View details for PubMedID 18628316
Ultrasound-guided percutaneous pancreatic tumor biopsy in pancreatic cancer: a comparison with metastatic liver tumor biopsy, including sensitivity, specificity, and complications
JOURNAL OF GASTROENTEROLOGY
2008; 43 (3): 225-232
Expression levels of insulin-like growth factor type 1 receptor (IGF-IR), epidermal growth factor receptor (EGFR), and HER2 expressions have been linked to clinical outcomes in several solid tumors. However, the clinical significance of these biomarkers in gastric cancer (GC) remains unclear. This study was designed to delineate the clinical implications of these three biomarkers in GC.The study group comprised 87 patients who underwent gastrectomy at National Cancer Center Hospital and subsequently received chemotherapy for recurrent or residual tumors. Using immunohistochemical techniques, we analyzed the expressions of IGF-IR, EGFR, and HER2 on formalin-fixed paraffin-embedded specimens of surgically removed primary tumors.IGF-IR expression (defined as >10% membranous staining) was found in 67 tumors (77%), EGFR expression in 55 (63%), and HER2 expression in 16 (18%). Positive coexpression of IGF-IR and EGFR was found in 48 tumors (55%), that of IGF-IR and HER2 in 16 (18%), and that of EGFR and HER2 in 13 (15%). Multivariate survival analysis showed that IGF-IR-positive expression [hazard ratio (HR) 2.14, 95% confidence interval (95% CI) 1.20-3.82; P = 0.01], performance status 1 or 2 (HR 1.83, 95% CI 1.15-2.91; P = 0.01), and diffuse type tumors (HR 1.71; 95% CI 1.08-2.70; P = 0.02) were significant predictors of poor survival.IGF-IR expression in surgical GC specimens, poor performance status, and diffuse type tumors are significant predictors of poor outcomes in patients with GC. Our data suggest that anti-IGF-IR strategies may prove valuable in such patients.
View details for DOI 10.1158/1078-0432.CCR-07-1898
View details for Web of Science ID 000256012700016
View details for PubMedID 18483367
Impacts of excision repair cross-complementing gene 1 (ERCC1), dihydropyrimidine dehydrogenase, and epidermal growth factor receptor on the outcomes of patients with advanced gastric cancer
BRITISH JOURNAL OF CANCER
2008; 98 (4): 832-839
The aims of this study were to investigate the diagnostic value and safety of ultrasound-guided percutaneous pancreatic tumor biopsy (pancreatic biopsy) in patients with suspected unresectable pancreatic cancer, and to compare the data with those obtained by metastatic liver tumor biopsy (liver metastases biopsy).Data were collected retrospectively from 388 patients (398 procedures) for whom a final diagnosis was available and who underwent ultrasound-guided pancreatic or liver metastases biopsy with a 21-gauge needle (core biopsy) or a 22-gauge needle (fine-needle aspiration biopsy: FNAB). The sensitivity, specificity, and accuracy of pancreatic and liver metastases biopsies were evaluated. Biopsy-related complications were collected and analyzed.Data from 271 pancreatic and 112 liver metastases biopsy procedures were available. For pancreatic core biopsy and FNAB, the sensitivity, specificity, and accuracy were 93%, 100%, and 93%, and 86%, 100%, and 86%, respectively, all of which were comparable to those of liver metastases biopsy. The complication rate in pancreatic biopsy was 21.4%, including a 4.4% incidence of post-biopsy ephemeral fever. The complication rate in liver metastases biopsy was 38.7%, including an 8.0% incidence of ephemeral fever. Fever and infection occurred more frequently among patients who underwent liver metastases biopsy (4.4% vs. 11%: P = 0.038). In pancreatic biopsy cases, a prebiopsy high serum total bilirubin level was a statistically significant predictor of ephemeral fever.Ultrasound-guided percutaneous pancreatic biopsy is an effective and safe modality for confirming the pathologic diagnosis in patients with unresectable pancreatic cancer.
View details for DOI 10.1007/s00535-007-2142-9
View details for Web of Science ID 000254456100007
View details for PubMedID 18373165
Clinical significance of insulin-like growth factor type 1 receptor and epidermal growth factor receptor in patients with advanced gastric cancer
2008; 74 (1-2): 76-83
Using laser-captured microdissection and a real-time RT-PCR assay, we quantitatively evaluated mRNA levels of the following biomarkers in paraffin-embedded gastric cancer (GC) specimens obtained by surgical resection or biopsy: excision repair cross-complementing gene 1 (ERCC1), dihydropyrimidine dehydrogenase (DPD), methylenetetrahydrofolate reductase (MTHFR), epidermal growth factor receptor (EGFR), and five other biomarkers related to anticancer drug sensitivity. The study group comprised 140 patients who received first-line chemotherapy for advanced GC. All cancer specimens were obtained before chemotherapy. In patients who received first-line S-1 monotherapy (69 patients), low MTHFR expression correlated with a higher response rate (low: 44.9% vs high: 6.3%; P=0.006). In patients given first-line cisplatin-based regimens (combined with S-1 or irinotecan) (43 patients), low ERCC1 correlated with a higher response rate (low: 55.6% vs high: 18.8%; P=0.008). Multivariate survival analysis of all patients demonstrated that high ERCC1 (hazard ratio (HR): 2.38 (95% CI: 1.55-3.67)), high DPD (HR: 2.04 (1.37-3.02)), low EGFR (HR: 0.34 (0.20-0.56)), and an elevated serum alkaline phosphatase level (HR: 1.00 (1.001-1.002)) were significant predictors of poor survival. Our results suggest that these biomarkers are useful predictors of clinical outcomes in patients with advanced GC.
View details for DOI 10.1038/sj.bjc.6604211
View details for Web of Science ID 000253219700023
View details for PubMedID 18231104
To better understand the clinical implications of insulin-like growth factor type 1 receptor (IGF-1R), epidermal growth factor receptor (EGFR) and HER2 expressions in gastric cancer (GC).The study group comprised 86 patients who received first-line chemotherapy for advanced GC at the National Cancer Center Hospital. Using laser-captured microdissection and a real-time RT-PCR assay, we quantitatively evaluated mRNA levels of IGF-1R, EGFR and HER2 in paraffin-embedded cancer specimens of surgically removed primary tumors.In univariate analysis of the study group as a whole, patients with low expression of both IGF-1R and EGFR (n = 13) had a significantly longer overall survival than the other patients (n = 51; median, 24.6 vs. 12.8 months; log-rank p = 0.013). Multivariate survival analysis demonstrated that high EGFR expression [hazard ratio, HR: 2.94 (95% confidence interval, CI: 1.40-6.17), p = 0.004] and poor performance status [HR: 1.96 (95% CI: 1.12-3.42), p = 0.018] were significant predictors of poor survival. In patients given first-line S-1 monotherapy (n = 29), low IGF-1R (p = 0.002) and low EGFR (p = 0.035) gene expression correlated with a better response, without a significant prolongation of survival.Our data warrant further investigations on the strategy of co-targeting IGF-1R and EGFR in GC.
View details for DOI 10.1159/000139127
View details for Web of Science ID 000256752300012
View details for PubMedID 18544998