Serum Biomarker For Cancer Research Articles

Gold interdigitated triple-microelectrodes for label-free prognosticative aptasensing of prostate cancer biomarker in serum

A simple, single-masked gold interdigitated triple-microelectrodes biosensor is presented by taking the advantage of an effective self-assembled monolayer (SAM) using an amino-silanization technique for the early detection of a prostate cancer's biomarker, the prostate-specific antigen (PSA). Unlike most interdigitated electrode biosensors, biorecognition happens in between the interdigitated electrodes, which enhances the sensitivity and limit of detection of the sensor. Using the Faradaic mode electrochemical impedance spectroscopy (EIS) technique to quantify the PSA antigen, the developed sensing platform demonstrates a logarithmic detection of PSA ranging from 0.5 ng/ml to 5000 ng/ml, an estimated LOD down to 0.51 ng/ml in the serum, and a good sensor's reproducibility. The sensor's detection range covers the clinical threshold value at 4 ng/ml and the crucial diagnosis ‘grey zone’ of 4–10 ng/ml of PSA in serum for an accurate cancer diagnosis. The selectivity test revealed an excellent discrimination of other competing proteins, with a recorded detection signals at 5 ng/ml PSA as high as 7-fold increase versus the human serum albumin (HSA) and 8-fold increase versus the human glandular kallikrein 2 (hK2). The stability test showed an acceptable stability of the aptasensor recorded at six (6) days before the detection signal started degrading below 10% of the peak detection value. The developed sensing scheme is proven to exhibit a great potential as a portable prostate cancer biosensor, also as a universal platform for bio-molecular sensing with the versatility to implement nanoparticles and other surface chemistry for various applications.

Serum biomarker panels for the diagnosis of gastric cancer.

Gastric cancer is a leading cause of mortality due to neoplastic disease. Although early detection of gastric cancers can decrease the mortality rate, it remains a diagnostic challenge because of the lack of effective biomarkers. In this study, fifteen gastric cancer patients and ten healthy subjects were recruited to assess novel serum biomarkers for gastric cancer using antibody microarray technology. ELISA was utilized to validate the antibody array results. As a result, compared to the controls, eleven cytokines were found to be significantly increased in gastric cancer, including interferon gamma receptor 1 (IFNGR1), neurogenic locus notch homolog protein 3 (Notch‐3), tumor necrosis factor receptor superfamily member 19L (TNFRSF19L), growth hormone receptor (GHR), signaling lymphocytic activation molecule family 8 (SLAMF8), folate receptor beta (FR‐beta), integrin alpha 5, galectin‐8, erythropoietin‐producing hepatocellular A1 (EphA1), epiregulin, and fibroblast growth factor 12 (FGF‐12) with P < 0.05. ELISA validation supported the results of the antibody array. More importantly, most of these eleven cytokines, including IFNGR1, TNFRSF19L, GHR, SLAMF8, FR‐beta, and integrin alpha 5 were discovered to be elevated in gastric cancer serum samples for the first time in this study, suggesting that these proteins may serve as novel biomarkers for the early diagnosis and prognosis determination of gastric cancer.

Simultaneous Measurement of 92 Serum Protein Biomarkers for the Development of a Multiprotein Classifier for Ovarian Cancer Detection.

The best known ovarian cancer biomarker, CA125, is neither adequately sensitive nor specific for screening the general population. By using a combination of proteins for screening, it may be possible to increase the sensitivity and specificity over CA125 alone. In this study, we used Proseek Multiplex Oncology II plates to simultaneously measure the expression of 92 cancer-related proteins in serum using proximity extension assays. This technology combines the sensitivity of the PCR with the specificity of antibody-based detection methods, allowing multiplex biomarker detection and high-throughput quantification. We analyzed 1 μL of sera from each of 61 women with ovarian cancer and compared the values obtained with those from 88 age-matched healthy women. Principle component analysis and unsupervised hierarchical clustering separated the ovarian cancer patients from the healthy, with minimal misclassification. Data from the Proseek plates for CA125 levels exhibited a strong correlation with clinical values for CA125. We identified 52 proteins that differed significantly (P < 0.006) between ovarian cancer and healthy samples, several of which are novel serum biomarkers for ovarian cancer. In total, 40 proteins had an estimated area under the ROC curve of 0.70 or greater, suggesting their potential to serve as biomarkers for ovarian cancer. CA125 alone achieved a sensitivity of 93.4% at a specificity of 98%. By adding the Oncology II values for five proteins to CA125 in a multiprotein classifier, we increased the assay sensitivity to 98.4% at a specificity of 98%, thereby improving the sensitivity and specificity of CA125 alone.

Proteomic Screening for Serum Biomarkers for Cervical Cancer and Their Clinical Significance.

BackgroundThe present study aimed to determine serum markers for cervical cancer (CC) and to provide valuable references for clinical diagnosis and treatment.Material/MethodsSerum samples were collected from age-matched healthy control women, and from female CC patients before and after surgery. Serum biomarkers were selected by comparing serum peptides profiles among the 3 groups by magnetic bead-based weak cation – exchange chromatography fractionation combined with matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Probable serum biomarkers for cervical cancer were then further identified by liquid chromatography-electrospray ionization-tandem mass spectrometry system and the identified proteins were verified by enzyme-linked immunosorbent assay (ELISA).ResultsThree peptide biomarkers were identified for distinguishing CC patients from normal individuals, and distinguishing preoperative CC patients from postoperative CC patients. Of these 3 identified protein peptide regions, 2 peptide regions – TKT (Peak 2, 2435.63 m/z, 499–524) and FGA (Peak 4, 2761.79 m/z, 603–629) – were identified as upregulated markers, and peptide region of APOA1 (Peak 9, 2575.3 m/z, 245–260) was identified as a downregulated biomarker in preoperative CC patients compared with healthy women.ConclusionsThe present study provides a new method for identifying potential serum biomarkers for CC patients.

Population Change of Myeloid and Lymphoid Cells Contributes to the Blood-Based Protein Biomarkers

Background: It has been observed that myeloid cells proportion increases, while lymphoid cells proportion decreases in blood of both inflammatory and cancer patients, which explains the serum concentration changes of many protein biomarkers in cancer patients versus normal controls. Methods: From literature, cancer serum and/or plasm protein biomarkers were collected and their expression patterns were examined in the microarray data of myeloid and lymphoid cells. Serum from 30 pancreatic ductal adenocarcinoma (PDAC) patients, 30 pancreatitis patients and 30 healthy volunteers, was studied using the enzyme-linked immunosorbent assay (ELISA). Findings: Among 45 protein biomarkers for PDAC, the genes expressing 15 proteins were significantly dysregulated in myeloid cells versus lymphoid cells and 13 of them, except APOA2 and THBS1, were dysregulated to the same directions as in PDAC patients versus normal controls (binomial test, P = 3.7E-3). A further comparative study was carried out on other cancers and showed significantly consistency (binomial test, P = 1.54E-02). Experimentally, we confirmed that APOA2 and THBS1 were significantly up-regulated in PDAC patients versus normal controls (Student's t test, P < 0.05) as in myeloid cells versus lymphoid cells, while there were no significant differences between PDAC and pancreatitis patients. Interpretation: We demonstrate that the ratio shifts of myeloid and lymphoid cells might lead the observed changes of many secreted protein biomarkers for cancers. Funding Statement: National Natural Science Foundation of China, Natural Science Foundation of Fujian Province, Education and Scientific Research Projects of Young and Middle-aged Teachers in Fujian Province and Fujian Medical University. Declaration of Interests: All authors declare no conflict of interest. Ethics Approval Statement: All subjects gave their informed consent for inclusion before they participated in the study. The study was approved by the Ethics Committee of Fujian Medical University and all experiments were performed in accordance with the approved guidelines and regulations.

The Evaluation of Serum Biomarkers for Non-small Cell Lung Cancer (NSCLC) Diagnosis.

Introduction: Lung cancer ranks top in the cause of cancer death globally. The identification of effective biomarkers is essential for non-small cell lung cancer (NSCLC) diagnosis.Methods: The expression levels of prolactin (PRL), CEA, and CYFR21 in serum were assayed by ELISA. The blood samples were attained from 44 NSCLC cases and 44 healthy controls. Logistic regression and receiver operating characteristic (ROC) analyses were applied to evaluate the diagnostic efficacy and create diagnostic mathematical models.Results: Serum PRL, CEA, and CYFR21 levels were significantly higher in patients with NSCLC than the healthy controls (all P-values <0.001). According to the model to predict NSCLC patients from the healthy controls, a combination of PRL, CEA, and CYFR21 biomarkers was more effective than individual biomarker alone, with AUC = 0.960 (95% CI: 0.921–0.999), sensitivity = 0.909, specificity = 0.955, positive predicted value = 0.952, and negative predicted value = 0.913.Conclusion: Prolactin can be used as a potential serum biomarker for the diagnosis of NSCLC. A panel of PRL, CEA, and CYFRA21 was found as promising serum biomarkers for the diagnosis of NSCLC with relatively high sensitivity and specificity.

Extracellular cyclic adenosine monophosphate-dependent protein kinase A autoantibody and C-reactive protein as serum biomarkers for diagnosis of cancer in dogs.

Protein kinase A, a cyclic adenosine monophosphate (AMP)‐dependent enzyme, normally exists within mammalian cells; however, in cancer cells, it can leak out and be found in the serum. Extracellular cyclic AMP‐dependent protein kinase A (ECPKA) has been determined to increase in the serum of cancer‐bearing dogs. However, there have been no reports in the veterinary literature on serum ECPKA autoantibody (ECPKA‐Ab) expression in dogs with cancer. The aim of this study was to evaluate ECPKA‐Ab and C‐reactive protein (CRP) as serum biomarkers for cancer in dogs. ECPKA‐Ab and CRP levels were detected by an enzyme‐linked immunosorbent assay in serum samples from dogs with malignant tumours (n = 167), benign tumours (n = 42), or non‐tumour disease (n = 155) and from healthy control dogs (n = 123). ECPKA‐Ab and CRP levels were significantly higher in the dogs with malignant tumours than in those with benign tumours or non‐tumour diseases, as well as in the healthy controls (P < 0.001, Kruskal‐Wallis test). There was a significant positive correlation between the neoplastic index, which was developed using ECPKA‐Ab and CRP levels, and the presence of cancer in dogs (P < 0.001); the area under the receiver‐operating characteristic curve was estimated to be >0.85 (P < 0.001). In conclusion, ECPKA‐Ab is a potential serum biomarker for a broad spectrum of cancers. Combined measurement of CRP and ECPKA‐Ab levels in serum improves the sensitivity and accuracy of a diagnosis of cancer in dogs.

PLOD3 promotes lung metastasis via regulation of STAT3

Procollagen-lysine, 2-oxoglutarate 5-dioxygenase (PLOD3), a membrane-bound homodimeric enzyme, hydroxylates lysyl residues in collagen-like peptides; however, its role in lung cancer is unknown. This study aimed to investigate the role of PLOD3 as a pro-metastatic factor and to elucidate the underlying mechanism. First, we experimentally confirmed the release of PLOD3 in circulation in animal models, rendering it a potential serum biomarker for lung cancer in humans. Thereafter, we investigated the effects of PLOD3 overexpression and downregulation on cancer cell invasion and migration in vitro and in vivo, using human lung cancer cell lines and a mouse tumor xenograft model, respectively. Further, PLOD3 levels were determined in lung tissue samples from lung cancer patients. Functional analyses revealed that PLOD3 interacts with STAT3, thereby expressing matrix metalloproteinases (MMP-2 and MMP-9) and with urokinase plasminogen activator (uPA) to enhance tumor metastasis. PLOD3 and the STAT3 pathway were significantly correlated in the metastatic foci of lung cancer patients; PLOD3–STAT3 levels were highly correlated with a poor prognosis. These results indicate that PLOD3 promotes lung cancer metastasis in a RAS-MAP kinase pathway-independent manner. Therefore, secreted PLOD3 serves as a potent inducer of lung cancer metastasis and a potential therapeutic target to enhance survival in lung cancer.

Olive fruit (Olea europaea L.): Chemopreventive effect in the rat model of hepatocellular carcinoma

Primary liver cancer or hepatocellular carcinoma (HCC) is an aggressive tumor with poor prognosis. Olea europaea L. commonly called olive contains diverse phytochemical constituents that have preventive effects against different diseases. The purpose of this investigation was to study the chemopreventive effect of aqueous extract of Olea europaea L. (AOE) against diethylnitrosamine (DEN)-induced HCC in rats. Olive-treated rats were pretreated with AOE intragastrically at three different doses two weeks prior to DEN injection that continued until 8 weeks. At three different time points, blood samples were collected for biochemical analysis. In the present investigation, a significant decrease in serum biomarkers of liver damage and cancer, including alfa-fetoprotein (AFP), gamma glutamyl transpeptidase (GGT), alanine transaminase (ALT), and aspartate transaminase (AST) were observed in AOE supplemented animals when compared to DEN-treated rats. In addition, AOE counteracted DEN-induced hepatic oxidative stress in rats illustrated by the restoration of reduced glutathione (GSH) and the reduction of lipid peroxidation in HCC rats. Furthermore, AOE prevented increase of relative liver weight as a prognostic marker in HCC. Finally, histopathological investigation in the liver sections confirmed the preventive effect of AOE against DEN-induced HCC. To conclude, olive has a preventive effect against DEN-induced hepatocarcinogenesis in rats.

Examining transcriptional changes to DNA replication and repair factors over uveal melanoma subtypes

BackgroundUncontrolled replication is a process common to all cancers facilitated by the summation of changes accumulated as tumors progress. The aim of this study was to examine small groups of genes with known biology in replication and repair at the transcriptional and genomic levels, correlating alterations with survival in uveal melanoma tumor progression. Selected components of Pre-Replication, Pre-Initiation, and Replisome Complexes, DNA Damage Response and Mismatch Repair have been observed.MethodsTwo groups have been generated for selected genes above and below the average alteration level and compared for expression and survival across The Cancer Genome Atlas uveal melanoma subtypes. Significant differences in expression between subtypes monosomic or disomic for chromosome 3 have been identified by Fisher’s exact test. Kaplan Meier survival distribution based on disease specific survival has been compared by Log-rank test.ResultsGenes with significant alteration include MCM2, MCM4, MCM5, CDC45, MCM10, CIZ1, PCNA, FEN1, LIG1, POLD1, POLE, HUS1, CHECK1, ATRIP, MLH3, and MSH6. Exon 4 skipping in CIZ1 previously identified as a cancer variant, and reportedly used as an early serum biomarker in lung cancer was found. Mismatch Repair protein MLH3 was found to have splicing variations with deletions to both Exon 5 and Exon 7 simultaneously. PCNA, FEN1, and LIG1 had increased relative expression levels not due to mutation or to copy number variation.ConclusionThe current study proposes changes in relative and differential expression to replication and repair genes that support the concept their products are causally involved in uveal melanoma. Specific avenues for early biomarker identification and therapeutic approach are suggested.

Abstract B20: Simultaneous measurement of 92 serum protein biomarkers for the development of a multiprotein classifier for ovarian cancer detection

Abstract Currently, there are no FDA-approved screening tools for detecting ovarian cancer in the general population. The two most common ovarian cancer biomarkers, CA125 and HE4, when used individually are neither adequately sensitive nor specific enough to be used for screening. We hypothesize that by using a combination of biomarkers for screening, it will be possible to increase the sensitivity and specificity over CA125 alone. Development of a blood-based assay for ovarian cancer detection could significantly improve the survival of patients if it identified cancers earlier. In this study, we used Proseek Multiplex Oncology II plates to simultaneously measure the expression of 92 cancer-related proteins in serum using proximity extension assays. This technology combines the sensitivity of the polymerase chain reaction with the specificity of antibody-based detection methods, allowing multiplex biomarker detection and high-throughput quantification. In a review of the literature, we found that 44 of these 92 proteins had been studied individually and were present at elevated levels in ovarian cancer samples (e.g., serum or tissues). This study is the first to examine the levels of 92 proteins simultaneously using a large cohort of ovarian cancer serum samples. We analyzed one microliter of serum from each of 60 women with late stage (Stages III and IV) high-grade serous ovarian cancer and compared the values obtained to those from 241 age-matched healthy women. The data from these 301 serum samples were normalized across four Proseek Multiplex Oncology II 96-well plates and across the different sources of sera. Principal component analysis and unsupervised hierarchical clustering separated patients into the two major groups of ovarian cancer and healthy, with minimal misclassification. Data from the Proseek plates for CA125 levels exhibited a strong correlation with previously measured clinical values for CA125 (correlation coefficient of 0.90, with a 95% confidence interval of 0.88-0.92). CA125 and HE4 were detected at low levels in healthy samples, while higher levels were found in the ovarian cancer cases. Overall, we identified 45 proteins that showed a significant difference (p &amp;lt; 0.001) between ovarian cancer and healthy samples, many of which could serve as novel serum biomarkers for ovarian cancer. In total, 46 proteins had an estimated area under the ROC curve (AUC) of 0.7 or greater. CA125 alone achieved a sensitivity of 0.92 and a specificity of 0.98. In addition, we generated a multiprotein classifier using 10 proteins (including CA125) that increased the sensitivity to 0.97, while holding the specificity fixed at 0.98. This 5% increase in sensitivity will have a significant effect on the number of women correctly identified when screening a large population. Our data demonstrate that the Proseek technology can replicate the results established by conventional clinical assays for known biomarkers, identify new candidate biomarkers, and improve the sensitivity and specificity of CA125 alone. Additional studies using a larger cohort of early-stage ovarian cancer patients will allow for validation of these biomarkers and lead to the development of a screening tool for detecting early-stage ovarian cancer in the general population. Citation Format: Amy P.N. Skubitz, Kristin L.M. Boylan, Kate Geschwind, Qing Cao, Timothy K. Starr, Melissa A. Geller, Robert C. Bast, Jr., Karen H. Lu, Joseph S. Koopmeiners. Simultaneous measurement of 92 serum protein biomarkers for the development of a multiprotein classifier for ovarian cancer detection. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr B20.

Abstract 1059: EpCAM differentially expressed on circulating and disseminated tumor cells promotes tumor metastasis in breast cancer patients

Abstract To investigate the role of EpCAM expressed on both circulating tumor cells (CTCs) in peripheral blood and disseminated tumor cells (DTCs) in bone marrow with respect to tumor metastases, the integrated subtraction enrichment and immunostaining-FISH (SE-iFISH) was performed to quantify, phenotypically and karyotypically characterize cytogenetically abnormal CTCs and DTCs expressing EpCAM in breast cancer patients. In a cohort of 34 recruited late stage patients, consisting of 32 metastatic and 2 non-metastatic subjects, the average number of detected CTCs in metastatic vs non-metastatic patients was 25 and 18 cells, respectively. Among those CTCs in metastatic subjects, 23.4% of them showed EpCAM+, whereas none of EpCAM+ CTCs was detected in non-metastatic patients. There were total of 49 CTC clusters detected in metastatic patients, and 33% of those clusters were EpCAM+, however, only 3 CTC clusters were found in non-metastatic patients, and none of them was EpCAM+. In the case of DTCs, the average number of 2224 and 364 DTCs per subject was detected in metastatic vs non-metastatic subjects, with EpCAM+ ratio of 66.53% in metastatic and 8% in non-metastatic patients. Analysis of DTC clusters indicated that the average of 226 and 21 DTC clusters/subject were identified in metastatic and non-metastatic patients, respectively, and metastatic subjects showed 62% of DTC clusters EpCAM+, whereas 37% of clusters were EpCAM+ in non-metastatic patients. Comparing to the breast cancer serum biomarker CA15-3 detected in 32 metastatic cancer patients, showing positivity (≥28 ng/ml) of only 41% (13 out of 32), both CTCs (≥5) and DTCs (≥5) had a positivity rate of 81 (26 out of 32) and 97% (31 out of 32), respectively. Obtained results indicated that EpCAM+ CTCs and DTCs are reliable markers in terms of evaluating therapeutic efficacy and predicting cancer patients' prognosis. Citation Format: Peter P. Lin, Junjian Li, Daisy Wang, Olivier Gires, Hongxia Wang. EpCAM differentially expressed on circulating and disseminated tumor cells promotes tumor metastasis in breast cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1059.

Biosensor with a 4-Channel Disposable Sensing Module Facilitating a Direct Sensitive Detection of Colon Cancer Biomarkers in Serum

Colon cancer has a survival rate of 90% or more when detected early, but the current clinical diagnostic methods of identifying colon cancer such as optical screening and biopsy for polyp samples have difficulties in early diagnosis due to low sensitivity and complex process. In this study, we developed a field-effect transistor(FET)-based biosensor with high sensitivity and specificity to quantitatively analyze colon cancer biomarkers in serum samples. Conventional FET-based sensors have limitations to detect target molecules in serum due to short Debye length and high noise signal from high concentrations of ions and many interfering species. In this study to circumvent these issues we propose a new dual-gate FET based biosensors featuring 4-channel disposable sensing module(DSM). We successfully demonstrated that our new biosensor can facilitate a fg/ml level detection of biomarkers in serum.

Aspartyl (Asparaginyl) ß hydroxylase (AABH) as a serum bio-marker for prostate cancer to identify prostate cancer, regardless of PSA level.

10 Background: Background: Prostate Specific Antigen (PSA) is a molecular marker of prostate cancer (PC). I most countries, standard of care suggests annual PSA testing in all men over 50 years of age; and for men at high risk, the test is recommended from 40 years of age. Due to low Specificity and low sensitivity of the PSA test, a large number of unnecessary biopsies occur every year. This low specificity can be due to the fact that PSA is found in both benign and malignant lesions of Prostate tissue. To date measurement of the percentage of free PSA, have resulted an small improvements in specificity of PSA testing. Thus, the development of a simple blood test to detect prostate cancer that exhibits higher specificity compared to PSA could have the potential of reducing biopsies performed due to false positive screening results and improve the quality of medical care. Methods: We have investigated the utility of aspartyl (asparaginyl) β-hydroxylase (AABH) as a prostate cancer biomarker. AABH has been detected by immunohistochemical staining (IHC) in a broad range of cancers including Prostate Adenocarcinoma through out the world. AABH have been detected by IHC in more than 97% of tumor specimens tested (n &gt; 200) while absent in normal and non cancer tissue. We have developed a sandwich ELISA test for the detection of AABH in serum samples. Preliminary results showed an accuracy of 91- 94% for detecting cancer patients from non-cancer patients in different types of Cancer. Here we have utilized this assay to detect AABH levels in the sera of patients diagnosed with Prostate cancer (biopsy proven, pre-treatment samples), compared to non-cancer individuals. Results: AABH was detected above a threshold level of 1.2 ng/mL in 91% of the sera of PC patients (n = 60), in all different stages and grades of Prostate Cancer. All non-cancer individuals (n = 30) had AABH values below the threshold. Further study of direct comparison of AABH to PSA levels (n = 4,000) is underway. Conclusions: Our data suggest that measurement of serum AABH levels may help to detect Prostate Cancer in early stage and potentially reduce the number of prostate biopsies performed due to increased high serum PSA.

PLAC1 as a serum biomarker for breast cancer.

Placental-specific protein 1 (PLAC1) is an X-linked trophoblast gene that is re-expressed in several malignancies, including breast cancer, and is therefore a potential biomarker to follow disease onset and progression. Sera from 117 preoperative/pretreatment breast cancer patients and 51 control subjects, including those with fibrocystic disease, were analyzed for the presence of PLAC1 protein as well as its expression by IHC in tumor biopsies in a subset of subjects. Serum PLAC1 levels exceeded the mean plus one standard deviation (mean+SD) of the level in control subjects in 67% of subjects with ductal carcinoma in situ (DCIS), 67% with HER2+ tumors, 73% with triple-negative cancer and 73% with ER+/PR+ tumors. Greater sensitivity was achieved using the mean+2 SD of control PLAC1 serum values, where the false positive rate was 3% and was exceeded by 38%, 40%, 60% and 43% of subjects with DCIS, HER2+, TNBC and ER+/PR+/HER2- tumors. PLAC1 was detected in 97% of tumor biopsies, but did not correlate quantitatively with serum levels. There was no significant correlation of serum PLAC1 levels with race, age at diagnosis, body mass index (BMI) or the presence of metastatic disease. It remains to be determined whether PLAC1 serum levels can serve as a diagnostic biomarker for the presence or recurrence of disease post-surgery and/or therapy.

Aspartyl (Asparaginyl) β-Hydroxylase AABH as a serum biomarker for colorectal cancer.

86 Background: Colorectal Cancer (CRC) is the third most common type of cancer diagnosed in the US and Canada. WHO, Canadian Cancer Society (CCS), the National Comprehensive Cancer Network (NCCN) and American Cancer Society (ACS) recommend that men and women begin CRC screening at age 50 or younger if at high risk. Recommended screening procedures: Annual occult fecal blood test (OFBT), a colonoscopy every 5 years, OFBT and colonoscopy every 5 years, or a colonoscopy every 10 years. According to The Surveillance, Epidemiology, and End Results (SEER) Program, only 39% of CRC are diagnosed in stage I, 36% are diagnosed in Stage II, 19% are diagnosed with metastasis. The corresponding 5-year survival rates are 89.8%, 67.7%, and 10.3%. Neither the CCS nor the ACS recommends a blood test be done as part of screening. This is due to the fact that, until now, there has not been a blood test with adequate sensitivity or specificity for screening. Methods: In this study we discovered that Aspartyl (Asparaginyl) β-Hydroxylase (AABH) measurement in serum can be used as an screening test for CRC. AABH has been detected by immunohistochemical staining (IHC) on the cell surface of different cancers including CRC. It has been detected by IHC in &gt; 97% of tumor specimens tested (n &gt; 200) but has not been found in tissue samples from normal individuals. Results: This observation and the observation that AABH is found in the serum of patients with cancer, but not in n0n cancer patients, led us to develop a Sandwich ELISA Assay to measure AABH in serum. In the current study we have quantified AABH levels in CRC patients and compared it with normal individual. Increased levels of AABH were found in the serum of 91.5% of patients with CRC in all different stages of Cancer (n = 60). In normal individuals, AABH was essentially undetectable in serum (n = 30). AABH was identified in serum from patients with CRC irrespective of cancer stage. All serum AABH levels for stages I, II, III and IV were more than 3.3 ng/mL. Conclusions: Thus, our data indicate that by measuring AABH in the serum, we should be able to detect CC at an earlier stage than it is currently detected, resulting to a much better 5-year survival for CC.

The Potential of Angiogenin as a Serum Biomarker for Diseases: Systematic Review and Meta-Analysis.

Background Angiogenin (ANG) is a multifunctional angiogenic protein that participates in both normal development and diseases. Abnormal serum ANG levels are commonly reported in various diseases. However, whether ANG can serve as a diagnostic or prognostic marker for different diseases remains a matter of debate. Methods Here, we performed a systematic review and meta-analysis of the literature utilizing PubMed, Web of Science, and Scopus search engines to identify all publications comparing plasma or serum ANG levels between patients with different diseases and healthy controls, as were studies evaluating circulating ANG levels in healthy populations, pregnant women, or other demographic populations. Results This study demonstrated that the serum ANG concentration in healthy populations was 336.14 ± 142.83 ng/ml and remained relatively stable in different populations and regions. We noted no significant differences in serum ANG levels between patients and healthy controls, except in cases in which patients suffered from cancer or cardiovascular diseases. The serum ANG concentrations were significantly higher in patients who developed colorectal cancer, acute myeloid leukemia, multiple myeloma, myelodysplastic syndromes, and heart failure than those in healthy controls. Conclusion ANG has the potential of being a serum biomarker for cancers and cardiovascular diseases.

Prognostic serum biomarkers for colorectal cancer – Review

Prognostic serum biomarkers for colorectal cancer – Review

Synthetic glycopeptides as a designated standard in focused glycoproteomics to discover serum cancer biomarkers.

Previous studies on the large-scale glycomics of more than 3500 human serum samples revealed that the serum glycoproteins of cancer patients often have more dominant and specific glycoforms, namely, branched tri- and tetra-antennary N-glycans, most cancer patient groups than normal control groups. We herein established an efficient synthetic protocol of glycopeptides having highly complicated N-glycan structures that may be generated by direct tryptic digestion of serum glycoproteins. A preliminary selected reaction monitoring (SRM) assay using the synthetic model glycopeptide 1, 40Ser-Val-Gln-Glu-Ile-Gln-Ala-Thr-Phe-Phe-Tyr-Phe-Thr-Pro-Asn-Lys-Thr-Glu-Asp-Thr-Ile-Phe-Leu-Arg63 having an asialo tri-antennary N-glycan at the Asn54 residue as a designated calibration standard allowed for the rapid and absolute quantitation of the tryptic fragment derived from the serum α1-acid glycoprotein carrying a focused N-glycoform of cancer patients and healthy controls in a range between 200 and 1600 fmole μL-1 without any enrichment process for the target glycoprotein.

Prostate tumors downregulate microseminoprotein-beta (MSMB) in the surrounding benign prostate epithelium and this response is associated with tumor aggressiveness.

Microseminoprotein-beta (MSMB) is a major secretory product from prostate epithelial cells. MSMB synthesis is decreased in prostate tumors in relation to tumor grade. MSMB levels are also reduced in the circulation and MSMB is therefore used as a serum biomarker for prostate cancer. We hypothesized that cancers induce a reduction in MSMB synthesis also in the benign parts of the prostate, and that the magnitude of this response is related to tumor aggressiveness. Reduced levels of MSMB in the circulation could therefore be a consequence of reduced MSMB expression not only in tumor tissue but also in the benign prostate tissue. MSMB expression was analyzed in prostatectomy specimens from 36 patients using immunohistochemistry and qRT-PCR. MSMB expression in the benign prostate tissue was analyzed in relation to Gleason score, tumor stage, and distance to the tumor. Furthermore, Dunning rat prostate tumors with different aggressiveness were implanted into the prostate of Copenhagen rats to study if this affected the MSMB expression in the tumor-adjacent benign rat prostate tissue. In prostatectomy specimens, MSMB expression was reduced in prostate tumors but also in the tumor-adjacent benign parts of the prostate. The reduction in tumor MSMB was related to tumor grade and stage, and the reduction in the benign parts of the prostate to tumor grade, stage, and distance to the tumor. Implantation of Dunning cancer cells into the rat prostate resulted in reduced MSMB protein levels in the tumor-adjacent benign prostate tissue. Rapidly growing and metastatic MatLyLu tumors had a more pronounced effect than slow-growing non-metastatic G tumors. Our data suggest that aggressive prostate tumors suppress MSMB synthesis in the benign prostate and that this could explain why serum levels of MSMB are decreased in prostate cancer patients. This study suggests that markers for aggressive cancer can be found among factors altered in parallel in prostate tumors and in the adjacent benign tissue.