Study on Urinary Candidate Metabolome for the Early Detection of Breast Cancer.

A metabolomic study for determination of certain urinary metabolomes, 1-methyladenosine (1-MA), 1-methylguanosine (1-MG), and 8-hydroxy-2' deoxyguanosine (8-OHdG) in urine specimens of breast cancer patients. The accuracy of these metabolites and their combined score with cancer antigen 15-3 (CA15-3) was developed to improve the early detection of breast cancer. This study recruited 52 healthy individuals, 47 benign breast tumors, and 167 malignant breast tumor patients. Urine samples were handled to adjust the creatinine concentrations to 8 mg/dL (0.7 mmol/L) and analyzed using GC-MS to detect and quantify the selected urinary metabolomes in urine samples of all participants. The accuracy of individual urinary metabolomes and their combination with CA15-3 were evaluated using multivariate statistical analysis. The cutoff value of CA15-3 was 32.5 U/mL. Cutoff values of 1-MA, 1-MG, and 8-OHdG were 2.19, 2.1, and 7.3 µmol/mmol creatinine, respectively. The concentrations of 1-MA, 1-MG, and 8-OHdG were significantly higher in breast cancer patients, especially in the early-stage. The combination of three urinary metabolomes with CA15-3 improves the diagnostic sensitivity of breast cancer. For the combined score, the area under the curve (AUC) value of combined score ranged from 0.820 to 0.950, with high accuracy, ranged from 77.0 to 95.5%. The most significant AUC (0.973), sensitivity (90.1%), selectivity (94.0%) was recorded at comparing the healthy control with the early-stage of malignant breast cancer. In conclusion, the combination of three urinary metabolomes with serum CA15-3 improves the diagnostic sensitivity of breast cancer.

Association of Polymorphism in Survivin Gene and the Risk of Liver Cancer Resulting from Hepatitis C Virus Among Egyptian Patients.

BACKGROUND: This study aims to investigate the relation between Survivin gene polymorphisms and the risk of Hepatocellular carcinoma (HCC) resulting from hepatitis C infection among the Egyptian population. METHODS: This prospective study was conducted on 164 patients, 57 patients were diagnosed with hepatitis C, where 57 were diagnosed with HCC in addition to 50 healthy volunteers as controls. Genotyping for Survivin rs1042489 and rs8073069 single nucleotide polymorphisms was carried out by the allelic discrimination Real-Time Polymerase Chain Reaction Single Nucleotide Polymorphisms genotyping technology. RESULTS: The results of Survivin rs1042489 polymorphism revealed that the TC and CC genotypes were significantly different between hepatocellular carcinoma patients (OR=15.5, 95%CI: 3.299-72.825,P<0.001), and controls (OR=44, 95%CI: 8.025-241.254, P<0.001). Furthermore, CC genotype was significantly different between cirrhotic and hepatocellular carcinoma patients (OR=19.2, 95%CI: 3.097-119.049, P=0.002). Moreover, the TC genotype shows a significant difference between controls and cirrhotic patients (OR=5.5, 95%CI: 2.111-14.328, P<0.001). However, when comparing TT genotypes, CC+TC genotypes results showed a significant association with increasing the risk of cirrhosis and hepatocellular carcinoma (OR=4.812, 95%CI: 1.893-12.233, P=0.001), (OR=21.607, 95%CI: 4.738-98.532, P<0.01), respectively. On the other hand, there was no significant difference among all studied groups for all genotypes regarding Survivin rs8073069. Also, the CC+GC genotype showed no significant association with increased risk of hepatocellular carcinoma (P=0.999) compared with the GG genotypes. CONCLUSION: The study indicates that functional Survivin rs1042489 polymorphism may contribute to the risk of hepatocellular carcinoma while Survivin rs8073069 polymorphism has no significant association with increased risk of hepatocellular carcinoma among the studied groups.

Development of novel score based on Angiogenic panel for accurate diagnosis of hepatocellular carcinoma among hepatitis C virus high-risk patients.

BACKGROUND/AIMS: Hepatocellular carcinoma (HCC) is a primary malignancy of the liver and a global health problem. It is often diagnosed at advanced stage where hopeless for effective therapies. Identification of more reliable biomarkers for early detection of HCC is urgently needed. HCC is identified with hyper-vascularity feature. Herein, we sought to develop a novel score based on the combination of the most significant angiogenic biomarkers with and routine laboratory tests for accurate detection of HCC. MATERIAL & METHODS: Angiopoietin II, copper, nitric oxide, albumin, platelets count and α-fetoprotein were assayed in HCC patients (75), liver cirrhosis patients (50) and healthy control (20). Areas under receiving operating curve (AUCs) were calculated and used for construction on novel score. A novel score named HCC-AngioScore = AFP (U/L) Albumin (g/dl) × 5.4 + Angiopoietin (ng/ml) × 0.001 + Copper (mg/dl) × (-0.004) + Platelets count (×109)/L × 0.003 + Nitric oxide (µ mol/L) × 0.27-36 was developed. HCC-AngioScore produce AUC of 0.919for differentiate patients with HCC from those with liver cirrhosis with sensitivity and specificity of a cut-off 0 (i.e., less than 0 the case is considered cirrhotic, whereas above 0 it is considered HCC. CONCLUSION: HCC-AngioScore could replace AFP during screening of HCV patients and early detection of HCC.

Development of a gas chromatography-mass spectrometry method for breast cancer diagnosis based on nucleoside metabolomes 1-methyl adenosine, 1-methylguanosine and 8-hydroxy-2'-deoxyguanosine.

Metabolomes are small molecule metabolites (<1000 Da) produced by cellular processes. Metabolomes are close counterparts to the genome, transcriptome and proteome. The aim of this study was to develop a method to detect and quantify candidate nucleoside metabolomes 1-methyl adenosine (1-MA), 1-methylguanosine (1-MG) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) in the urine of patients with breast cancer using gas chromatography-mass spectrometry (GC-MS). The method was applied to urine specimens from patients with breast cancer (n = 56) and benign breast tumors (n = 22), as well as from healthy females (n = 20). The relative standard deviations of precision and repeatability analysis were <10%, and recoveries ranged from 88.5 to 105.6%. Limits of detection were 0.014, 0.012, and 0.018 mg/L for 1-MA, 1-MG and 8-OHdG, respectively. The lower limits of quantitation were 0.056, 0.048 and 0.072 mg/L, respectively. There were significant differences in concentrations of candidate metabolomes between patients with cancer and the healthy individuals, especially for those in the early stages of the disease (p < 0.001). No significant differences were observed between the benign and healthy groups. In conclusion, a reliable GC-MS method for the detection and quantification of 1-MA, 1-MG, and 8-OHdG metabolomes in urine has been developed.

Association of monoallelic MUTYH mutation among Egyptian patients with colorectal cancer.

Colorectal cancer (CRC) is a heterogeneous triat that involves both environmental and genetic factors. Genetic mutations of MUTYH (p.Y179C and p.G396D) have been reported to be associated with increased risk of CRC among several ethnic populations. The aim of this work is to assess the association of the monoallelic MUTYH mutations (p.Y179C and p.G396D) with increased risk of CRC among Egyptian patients. This study included 120 unrelated CRC Egyptian patients who were compared with 100 healthy controls from the same locality. For all individuals, DNA was genotyped for MUTYH p.Y179C and MUTYH p.G396D mutations using the T-ARMS-PCR technique. The frequencies of monoallelic MUTYH mutations showed a strong association with the increased risk of CRC among Egyptian patients compared with controls (12.5 vs. 4.0 %, OR = 3.49, 95 % CI = 1.12-10.90, P = 0.03). Moreover, the frequency of MUTYH p.Y179C mutation was noted to be significantly higher among CRC patients compared to controls rather than MUTYH p.G396D mutation. Interestingly, CRC patients with tumors in the right side colon showed an evidence for association with the MUTYH p.Y179C mutation compared with tumors in the left side colon (p = 0.01). MUTYH p.Y179C mutation was associated with an increased risk of CRC among Egyptian patients rather than MUTYH p.G396D mutation.

Association of APC I1307K and E1317Q polymorphisms with colorectal cancer among Egyptian subjects.

Colorectal cancer is a multifactorial disease that involves both environmental and genetic factors. The gene encoding adenomatous polyposis coli (APC) has been reported to be associated with colorectal cancer (CRC) risk in several ethnic populations. The aim of this work is to assess the association of the APC I1307K and E1317Q polymorphisms with CRC risk among Egyptian subjects. This study included 120 unrelated CRC Egyptian patients who were compared to 100 healthy controls from the same locality. For all subjects, DNA was genotyped for APC I1307K and E1317Q polymorphisms using the PCR-ARMS technique. The frequency of APC I1307K carrier (TA+AA genotypes) was noted to be significantly higher among cases with CRC compared to controls (18.3 vs. 9.0 %, OR 2.58, 95 % CI 1.09-6.09, p = 0.03). Also the frequency of the APC I1307K A allele was significantly higher among cases compared to controls (10.4 vs. 4.5 %, OR 2.47; 95 % CI 1.12-5.42, p = 0.03). On the contrast, the frequencies of APC E1317Q GC genotype and C allele showed no significant difference among CRC patients compared to controls (3.3 vs. 2.0 %, OR 1.69; 95 % CI 0.30-9.42, p = 0.69 and 2.1 vs. 1.0 %, OR 2.11; 95 % CI 0.40-10.97, p = 0.46, respectively). Cases of the APC I1307K and E1317Q carriers (TA+AA and GC) showed no significant difference compared to those with I1307K and E1317Q non-carriers (TT and GG) regarding their clinical and laboratory markers. APC I1307K variant was associated with an increased risk of CRC among Egyptian subjects.

Development of a novel score for early detection of hepatocellular carcinoma among high-risk hepatitis C virus patients.

Hepatocellular carcinoma (HCC) is often diagnosed at advanced stage where effective therapies are lacking. Identification of new scoring system is needed to discriminate HCC patients from those with chronic liver disease. Based on the link between vascular endothelial growth factor (VEGF) and HCC progression, we aimed to develop a novel score based on combination of VEGF and routine laboratory tests for early prediction of HCC. VEGF was assayed for HCC group (123), liver cirrhosis group (210), and control group (50) by enzyme-linked immunosorbent assay (ELISA). Data from all groups were retrospectively analyzed including α-fetoprotein (AFP), international normalized ratio (INR), albumin and platelet count, transaminases, and age. Areas under receiving operating curve (ROC) were used to develop the score. A novel index named hepatocellular carcinoma-vascular endothelial growth factor score (HCC-VEGF score) = 1.26 (numerical constant + 0.05 × AFP (U l(-1)) + 0.038 × VEGF (ng ml(-1)) + 0.004 × INR - 1.02 × albumin (g l(-1)) - 0.002 × platelet count × 10(9) l- (1) was developed. HCC-VEGF score produce area under ROC curve of 0.98 for discriminating HCC patients from liver cirrhosis with sensitivity of 91 % and specificity of 82 % at cutoff 4.4 (i.e., less than 4.4 considered cirrhosis and greater than 4.4 considered HCC). Hepatocellular carcinoma-VEGF score could replace AFP in HCC screening and follow up of cirrhotic patients.

A novel discriminant score based on tumor-associated trypsin inhibitor for accurate diagnosis of metastasis in patients with breast cancer.

Invasion and metastasis of solid tumors require proteolytic enzymes for degradation of the basal membrane and extracellular matrix. Currently, there are no reliable methodologies to predict the risk for metastatic disease. In this context, our aim has been focused on the development of a noninvasive score based on tumor-associated trypsin inhibitor (TATI) for the assessment of metastasis in patients with breast cancer. TATI, trypsin, and soluble epidermal growth factor receptor (sEGFR) were assayed by enzyme-linked immunosorbent assay. CA 15.3 serum level was assayed by microparticle enzyme immunoassay in 265 patients with breast cancer. Statistical analyses were performed by logistic regression and receiver operating characteristic analysis curves. Using multivariate discriminant analysis, a score is selected based on absolute values of the four biochemical markers: TATI-metastatic breast cancer score (TATI-MBCS) = [0.03 × CA 15.3 (U/L) + 0.039 × TATI (ng/ml) + 0.04 × trypsin (ng/ml) + 0.023 × sEGFR (ng/ml) - 6.49 (numerical constant)]. This function correctly classified 84% of metastatic breast cancer at cutoff value = 0.62 (i.e., greater than 0.62 indicates patients with metastatic breast cancer and less than 0.62 indicates patients with nonmetastatic breast cancer). In conclusion, TATI-MBCS is a novel, noninvasive, and simple score which can be applied to discriminate patients with metastatic breast cancer.

Development of a novel metastatic breast cancer score based on hyaluronic acid metabolism.

Tumor metastasis involves the dissemination of malignant cells into the basement membrane, and the vascular system contributes to the circulating pool of these markers. In this context, our aim has been focused on the development of a non-invasive score based on degradation of the backbone of glycosaminoglycans of the extracellular matrix; namely hyaluronic acid (HA), for the assessment of metastasis in patients with breast cancer. HA level was determined by enzyme-linked immunosorbent assay; CA 15.3 was determined by microparticle enzyme immunoassay; hyaluronidase, N-acetyl-ß-D-glucosaminidase, ß-glucuronidase, glucuronic acid, and glucosamine were assayed by standard colorimetric techniques in 217 patients with breast cancer. Statistical analyses were performed by logistic regression and receiver-operating characteristic analysis curves. The multivariate discriminant analysis selects a score based on absolute values of the six biochemical markers: metastatic breast cancer score (MBCS) = [1.04 (Numerical constant) + 0.003 × CA 15.3 (U/l) + 0.001 × HA (ng/ml) + 0.004 × hyaluronidase (mg N-acetyl-ß-D-glucosamine/ml/18 h) + 0.001 × N-acetyl-ß-D-glucosaminidase (µmol/ml/min) + 0.026 × glucuronic acid (ng/ml) + 0.003 × glucosamine (µg/dl)]. This function correctly classified 87 % of metastatic breast cancer at cut-off value = 0.85 (i.e., great than 0.85 indicates patients with metastatic breast cancer and less than 0.85 indicates patients with non-metastatic breast cancer). MBCS is a novel, non-invasive, and simple score which can be applied to discriminate patients with metastatic breast cancer.

Discriminant function based on parameters of hyaluronic acid metabolism and nitric oxide to differentiate metastatic from non-metastatic colorectal cancer patients.

Colorectal cancer (CRC) is one of the most common causes of cancer-related deaths worldwide. Because there is currently no useful serological marker for metastatic colorectal cancer, the search for simple biomarkers for colorectal cancer diagnosis and prognosis is needed. Hyaluronic acid level was determined by ELISA; in addition to its degrading enzymes, degradation products and nitric oxide were determined by standard techniques in 185 CRC patients with and without metastases. Statistical analyses were performed by logistic regression and receiver-operating characteristic (ROC) curves. The multivariate discriminate analysis (MDA) selects a function based on absolute values of six biochemical markers; score = [-0.62 (numerical constant) + hyaluronic acid (pg/l) × 0.002 + hyaluronidase (mg N-acetyl glucosamine/ml/18 h) × 0.009-ß-glucuronidase (µmol/ml/min) × 0.07 + N-acetyl-ß-D-glucosaminidase (µmol/ml/min) × 0.019-glucuronic acid (µg/dl) × 0.001 + nitric oxide (µmol/l) × 0.01]. The selected MDA function correctly classified 92% of the metastatic CRC patients at a discriminate cut-off score = 0.24 (i.e., less than 0.24 indicated patients with non-metastatic colon cancer, and greater than 0.24 indicated patients with metastatic colon cancer with high degrees of sensitivity (100%) and specificity (93%)). The positive predictive and negative predictive values were also high (81% and 85%, respectively). Colorectal cancer patients can be simply and efficiently classified into metastatic or non-metastatic using their MDA score.