Non-invasive Biomarkers For Early Diagnosis Research Articles

Serum bile acids profiling by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application on pediatric liver and intestinal diseases.

Background A method for bile acid profiling measuring 21 primary and secondary bile acids in serum samples was developed and validated with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample preparation included spiking with internal standards followed by protein precipitation, centrifugation, drying under nitrogen gas and reconstitution. Extracted samples were injected onto a Phenomenex Kinetex C18 column (150 × 4.60 mm, 2.6 μm). Methods Data was collected with LC-MS/MS operated in negative ion mode with multiple reaction monitoring (MRM) and single reaction monitoring (SRM). The analytical run time was 12 min. Results The method showed excellent linearity with high regression coefficients (>0.99) over a range of 0.05 and 25 μM for all analytes tested. The method also showed acceptable intra-day and inter-day accuracy and precision. As a proof of concept, the analytical method was applied to patients with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), biliary atresia (BA), and necrotizing enterocolitis (NEC), and distinct bile acids profiles were demonstrated. Conclusions The method could be poised to identify possible biomarkers for non-invasive early diagnosis of these disorders.

Role of Plasma miRNA-501 Expression Profile as a Marker of Hepatocellular Carcinoma (HCC) Progression With Hepatitis C Virus Infection

Abstract Introduction HCV is a health problem that confronts many countries in the world. Those patients will develop complications like cirrhosis and HCC, which is one of the most common cancers in the world, especially in Egypt, and considered the third leading cause of death worldwide. The prognosis of HCC is still dismal due to the late diagnosis. miRNAs are small, short noncoding RNAs, which have roles in the diagnosis of HCC. In our study, we focus on biological aspects of miRNAs. We report that miR-501 is strongly expressed and observed in the process of HCC development. miR-501 regulation is important as an oncofetal relevant to the diagnosis of HCC. Method This study was conducted on 100 adult patients; 25 patients were positive for anti-HCV and 25 patients were negative for HCV and enrolled as a control group. Patients were categorized into three groups: fibrosis (n = 25), CHC (n = 25), and HCC (n = 25) related to HCV evident by CT abdomen. All patients and controls were subjected to full clinical assessment and laboratory investigation. Blood (8 mL) was withdrawn from subjects, and 3 mL was collected in EDTA tubes for processing total RNA extraction and miRNA. The remaining 5 mL was left for determination of biochemical parameters. miRN-501 expression levels were determined by RT-PCR. Results The data revealed a significant increase in levels of AST, ALT, ALP, and CBC in both HCC and CHC groups compared to controls. The results of miRNA expression showed that miR-501 was higher in the HCC group than non-HCC group at P1.1. Conclusion miR-501 can be used as a noninvasive biomarker for early diagnosis of HCC among patients with HCV on account of its affectability for HCC.

Application and clinical significance of exosomes in hepatocellular carcinoma

Exosomes are produced by multivesicular bodies, they are extracellular lipid nano-vesicles carrying a variety of bioactive substances, containing lipids, RNA, DNA and proteins. Exosomes produced by tumor cells carry important information about tumor cells. Exosomes produced by hepatocellular carcinoma can participate in the formation of a microenvironment suitable for the proliferation, growth, invasion, metastasis and drug resistance of tumor cells.Meanwhile, circulating exosomes can be used as non-invasive biomarkers for early diagnosis of tumors and as drug delivery vectors for treatment.This review summarizes the characteristics of exosomes in the growth, invasion and metastasis, immune escape, and drug resistance biology of hepatocellular carcinoma, as well as the potential clinical application of early detection and treatment. Key words: Exosomes; Biological markers; Carcinoma, hepatocellular; Therapeutic potential

Auditory Brainstem Dysfunction, Non-Invasive Biomarkers for Early Diagnosis and Monitoring of Alzheimer's Disease in Young Urban Residents Exposed to Air Pollution.

Alzheimer's disease (AD) is a biological construct defined by abnormal deposits of hyperphosphorylated tau and amyloid-β. The 2050 projection for AD in the USA is 14 million. There is a strong association between AD, air pollution, and traffic. Early diagnosis is imperative for intervention in the initial disease stages. Hearing and, specifically, the ability to encode complex sounds are impaired in AD. Nuclei in the auditory brainstem appear to be sensitive to neurodevelopmental and neurodegenerative disorders. Specifically, sustained exposure to air pollution is harmful to the brainstem; young residents of Metropolitan Mexico City (MMC) exposed to fine particulate matter and combustion-derived nanoparticles develop AD pathology in infancy. MMC clinically healthy children and teens have significant central delays in brainstem auditory evoked potentials (BAEPs). Herein, we review evidence that the auditory pathway is a key site of AD early pathology associated with air pollution and is significantly involved in AD patients. We strongly suggest electrophysiological screening, including BAEPs, be employed to screen individuals for early delays and to monitor progressive decline in patients diagnosed with mild cognitive impairment and AD. Understanding auditory dysfunction in early AD in pediatric and young adult populations may clarify mechanisms of disease progression. Air pollution is a risk factor for the development of AD and as the number of Americans with AD continues to grow without a cure, we need to focus on preventable, early causes of this fatal disease and intervene appropriately.

Measurement of urinary kininogen (KNG) fragments as a noninvasive tool for early diagnosis of pancreatic cancer (PaCa).

209 Background: There is a clear need to identify a non-invasive biomarker for early diagnosis of PaCa, in order to improve the overall survival of PaCa patients. Secretion of large amounts of proteases is a hallmark of PaCa, which results in an abundance of protease-induced cleavage products being excreted in the urine. This has led to speculation that measurement of PaCa-specific fragments in the urine might be useful as a tool for discrimination between PaCa patients and healthy controls. Herein, we introduce urinary KNG fragments as a promising biomarker for early diagnosis of PaCa. Methods: Urine samples were collected from PaCa patients and healthy volunteers, with the written informed consent, from January 2014 to July 2016. Urinary protein tryptic fragments derived from protein C-termini were measured using isobaric tags (iTRAQ) for their relative quantitation, and the diagnostic ability of the urinary levels of these fragments was evaluated by receiver operating characteristic (ROC) curve analysis. The fragments which showed an area-under-the-curve (AUC) of over 0.8 were selected as candidate fragments for further validation by the multiple-reaction-monitoring technique (MRM) combined with high-speed liquid chromatography. The urinary level of each candidate fragment was quantified by MRM, and the diagnostic capability of each for discriminating PaCa patients from healthy controls was evaluated by ROC curve analysis. Results: Urine samples of 39 PaCa patients (7 resectable, 32 unresectable) and 42 healthy controls were examined by iTRAQ to find 12,783 fragments. ROC curve analysis was carried out to select two candidate fragments (fragments A, B), both of which turned out to be KNG cleavage products. The urinary levels of the two fragments were measured in 23 resectable PaCa, 118 unresectable PaCa patients, and 42 healthy volunteers using high-speed-LC-/MRM. The AUCs of serum CA19-9 and urinary levels of fragments A and B for discriminating patients of PaCa from healthy controls were 0.89, 0.81 and 0.70, respectively. Conclusions: Urinary KNG fragments showed favorable diagnostic capability and were considered as promising, noninvasively measurable biomarkers of PaCa.

Lnc-TCL6 is a Potential Biomarker for Early Diagnosis and Grade in Liver Cirrhosis Patients

Background: Long non-coding RNAs (LncRNAs) have been proved to participate in the pathogenesis or as biomarkers in many diseases. The study aimed to identify some sensitive and noninvasive biomarkers for early diagnosis and grade in liver cirrhosis (LC) patients, especially in hepatitis B liver cirrhosis patients. Methods: LncRNAs were screened in whole blood samples by CapitalBiotech microarray in both healthy controls and LC patients to discover candidate biomarkers, and candidate biomarkers were subjected to training using qRT-PCR for the quantification of their expression levels in independent whole blood sample sets, including healthy controls, hepatitis B virus carriers (HBV carrier), chronic hepatitis B (CHB), and LC. Ultimately, an excellent biomarker was validated by Mann-Whitney test and ROC analysis for LC. Findings: Lnc-TCL6 was identified as a high diagnostic accurate biomarker for LC in Child-Pugh class C patients compared to healthy controls, CHB and HBV carrier (AUC=0.719, 0.815, 0.797, respectively). Meanwhile, it showed a favorable specificities and sensitivities in Child-Pugh A, B, C grade in LC patients (AUC=0.711, 0.724, 0.837, respectively). Compared to healthy controls, the expression of Lnc-TCL6 was obviously upregulated in Child-Pugh class A (4.07±3.28 vs 2.89±3.16, P=0.014). Conversely, the Lnc-TCL6 was significantly downregulated in Child-Pugh class C patients (1.19±2.09 vs 2.89±3.16, P<0.01). Interpretation: Lnc-TCL6 is a potential biomarker for early diagnosis and grade in LC patients, and a reliable predictor for poor prognosis in Child-Pugh class C LC patients. Funding Statement: This study was supported by National Natural Science Foundation of China (U1501224), the Science and Technology Developmental Special Foundation of Guangdong Province (2017B020226003), the Guangzhou City Science and Technology Program (201604020118), and the Projects of Guangzhou City Health Care Collaborative Innovation (201604020002). Declaration of Interests: The authors declare no competing interests. Ethics Approval Statement: The protocol for this clinical trial was approved by the institutional review boards or ethics committees from our hospital and written consent for the information to be stored in the hospital database and used for research was obtained from the study participants.

A fingerprint of amyloid plaques in a bitransgenic animal model of Alzheimer's disease obtained by statistical unmixing analysis of hyperspectral Raman data.

The global prevalence of Alzheimer's disease (AD) points to endemic levels, especially considering the increase of average life expectancy worldwide. AD diagnosis based on early biomarkers and better knowledge of related pathophysiology are both crucial in the search for medical interventions that are able to modify AD progression. In this study we used unsupervised spectral unmixing statistical techniques to identify the vibrational spectral signature of amyloid β aggregation in neural tissues, as early biomarkers of AD in an animal model. We analyzed spectral images composed of a total of 55 051 Raman spectra obtained from the frontal cortex and hippocampus of five bitransgenic APPswePS1ΔE9 mice, and colocalized amyloid β plaques by other fluorescence techniques. The Raman signatures provided a multifrequency fingerprint consistent with the results of synthesized amyloid β fibrils. The fingerprint obtained from unmixed analysis in neural tissues is shown to provide a detailed image of amyloid plaques in the brain, with the potential to be used as biomarkers for non-invasive early diagnosis and pathophysiology studies in AD on the retina.

Light-Induced Length Shrinkage of Rod Photoreceptor Outer Segments.

PurposeThis study was designed to verify light-induced outer segment (OS) length shrinkage of rod photoreceptors and to characterize its anatomic source at disc-level resolution.MethodsFrog (Rana pipiens) retinas were used for this study. Time-lapse light microscopy of freshly isolated OSs was employed to test transient rod OS changes at 10 ms temporal resolution. Histological light microscopy of dark- and light-adapted retinas was used to confirm light-induced rod OS length changes; and transmission electron microscopy (TEM) was used to quantify light-driven structural perturbation of rod OSs at disc level resolution.ResultsTime-lapse light microscopy images verified transient length shrinking responses in freshly isolated rod OSs. Histological light microscopy images confirmed reduced rod OS lengths in light-adapted retinas, compared to that of dark-adapted retinas. TEM images disclosed shortened inter-disc distances in light-adapted retinas compared to dark-adapted retinas.ConclusionsLight-induced rod OS length shrinkage was confirmed using time-lapse light microscopy of isolated rod OSs and histological light microscopy of dark- and light-adapted retinas. TEM revealed that the rod OS length shrinkage was correlated to the light-driven decrease of the space between individual discs, not the disc thickness itself.Translational RelevanceLight-induced transient rod response promises a noninvasive biomarker for early diagnosis of age-related macular degeneration and retinitis pigmentosa, in which the rod photoreceptors are known to be more vulnerable than cone photoreceptors.

New Insights in Bladder Cancer Diagnosis: Urinary miRNAs and Proteins.

Bladder cancer is the 10th-most common cancer worldwide. The diagnosis and follow-up of patients require costly invasive methods and due to these expenses, bladder cancer continues to be one of the expensive malignancies. Early diagnosis is crucial in bladder cancer as it is in other cancers; therefore, non-invasive biomarkers for early diagnosis are very important. In this review, we aimed to focus on the most recent investigations on potential urinary micro RNA (miRNA) and protein biomarkers for bladder cancer diagnosis and their associated pathways. Studies performed by different groups were compiled and the biomarker properties of various proteins and miRNAs in the urine of bladder cancer patients were evaluated. Key studies were obtained by searching keywords “bladder cancer, urinary miRNA, urinary protein, urinary biomarker”. Targets and the pathways of the miRNAs and proteins were analyzed according to mirBase Catalogue and Panther Database. The major pathways that are targeted by aberrantly expressed miRNAs are Cholecystokinin receptor (CCKR), p53, Wnt signaling pathway, and feedback loops. We hereby conclude that urinary micro RNAs and proteins are promising candidates for bladder cancer diagnosis. It should be noted that urine collection, storage conditions, choice of fraction, and normalization strategies should be standardized.

Analysis of the differential urinary protein profile in IgA nephropathy patients of Uygur ethnicity

BackgroundIgA nephropathy (IgAN) is one of the most common forms of idiopathic glomerular diseases and might lead to end-stage kidney disease. Accurate and non-invasive biomarkers for early diagnosis are required for early intervention and consequent therapy for IgAN patients. Because variance in the disease incidence and predisposing genes of IgAN has been detected among different ethnicities, the ethnicity factor should be considered in IgAN biomarker discovery. The differences in the protein profiles and pathological mechanisms of IgAN in patients of Uygur ethnicity need to be clearly illustrated.MethodsIn this study, we used urinary proteomics to discover candidate biomarkers of IgAN in patients of Uygur ethnicity. The urinary proteins from Uygur normal control and Uygur IgAN patients were extracted and analyzed using 2D-LC-MS/MS and isobaric tags for relative and absolute quantitation (iTRAQ) analysis.ResultsA total of 277 proteins were found to be differentially represented in Uygur IgAN compared with the respective normal controls. The bioinformatics analysis revealed that the immune response, cell survival, and complement system were activated in Uygur IgAN. Many differentially expressed proteins were found to be related to nephropathy and kidney injuries. Four candidate biomarkers were validated by Western blot, and these results were consistent with the iTRAQ results. ICAM1, TIMP1, SERPINC1 and ADIPOQ were upregulated in Uygur IgAN. Bioinformatic analysis revealed that the increase of ICAM1 and TIMP1 might be caused by IgAN, but the increase of SERPINC1 and ADIPOQ might be caused by proteinuria. SERPINC1 and ICAM1 were identified as the candidate biomarkers with excellent area-under-the-curve (AUC) values (0.84) for distinguishing Uygur IgAN from normal controls.ConclusionsUsing urinary proteomic analysis, we identified several candidate biomarkers for IgAN in patients of Uygur ethnicity. These results will prove helpful for exploring the pathological mechanism of IgAN in patients of Uygur ethnicity and for developing better treatments for these patients.

Low-level serum miR-24-2 is associated with the progression of colorectal cancer

colorectal cancer (CRC) is the second leading cause of cancer and cancer-related death in the world. Noninvasive biomarkers for early diagnosis of CRC are highly demanded. The up-regulation of specific microRNAs (miRNAs) in serum has been considered a promising biomarker of CRC and miR-24-2 may be a potential biomarker in the diagnosis the progression of CRC. Sixty-eighty healthy subjects and 228 CRC patients were divided into six groups: control group, CRC 0, CRC I, CRC II, CRC III, CRC IV and CRC V. Serum level of miR-24-2 was measured by real-time qPCR. Serum lipid profiles and oxidative-related molecules were also measured. Serum levels of miR-24-2 in CRC patients were significantly higher than healthy subjects (p< 0.05). In addition, the expression level of the miR-24-2 was decreased with the progression of CRC and reached the lowest level in CRC V. Spearman Rank Correlation analysis showed that miR-24-2 level was negatively related to the levels of superoxide dismutase (SOD) and reduced glutathione (GSH), aspartate transaminase (AST), alanine transaminase (ALT), cholesterol and triglyceride (p< 0.05). Serum miR-24-2 is a potential negative biomarker in the diagnosis of the progression of CRC patients and associated with biochemical indices.

Plasma Exosomal Long Non-Coding RNAs Serve as Biomarkers for Early Detection of Colorectal Cancer

Background/Aims: Colorectal cancer (CRC) is the third most commonly diagnosed malignancy and the second leading cause of cancer-related deaths worldwide. Thus, methods for early diagnosis of CRC are urgently needed. We aimed to identify potential long non-coding RNAs (lncRNAs) in circulatory exosomes that may serve as biomarkers for the detection of early-stage CRC. Methods: Exosomes from the plasma of CRC patients (n = 50) and healthy individuals (n = 50) were isolated by ultracentrifugation, followed by extraction of total exosomal RNAs using TRIzol reagent. Microarray analysis was used for exosomal lncRNA profiling in the two groups, and real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to determine the expression level of lncRNAs in all patients and healthy subjects. Results: The expression of six lncRNAs (LNCV6_116109, LNCV6_98390, LNCV6_38772, LNCV_108266, LNCV6_84003, and LNCV6_98602) was found to be significantly up-regulated in CRC patients compared with that in healthy individuals by qRT-PCR. The receiver operating characteristic curve was used to verify their diagnostic accuracy. The values of the area under the curve for these lncRNAs were 0.770 (LNCV6_116109), 0.7500 (LNCV6_98390), 0.6500 (LNCV6_38772), 0.6900 (LNCV_108266), 0.7500 (LNCV6_84003), and 0.7200 (LNCV6_98602). Conclusion: Our study suggested that the expression of these six exosomal lncRNAs (LNCV6_116109, LNCV6_98390, LNCV6_38772, LNCV_108266, LNCV6_84003, and LNCV6_98602) was significantly up-regulated in the plasma of CRC patients, and that they may serve as potential non-invasive biomarkers for early diagnosis of CRC.

The Role of Beta-7 Integrin and Carbonic Anhydrase IX in Predicting the Occurrence of de Novo Nonalcoholic Fatty Liver Disease in Liver Transplant Recipients.

We consider very important for our patients the development of new non-invasive biomarkers for early diagnosis of NAFLD and NASH, as the "gold-standard" of liver biopsy is not easily accepted in clinical practice. Also NAFLD and NASH are dynamic processes that need prospective and repeated assessments, a need that cannot be met by the classical liver biopsy.

Plasma microRNA192 in combination with serum CA19-9 as non-invasive prognostic biomarker in periampullary carcinoma.

In this study, circulating microRNAs (miRNAs) are being investigated as non-invasive biomarkers for early diagnosis and prognosis of human cancers. Since the prognosis for pancreatobiliary subtype of periampullary carcinoma is poor, we assessed the prognostic relevance of miRNAs in combination with CA19-9 as noninvasive biomarker in periampullary carcinoma. Circulating miRNAs in plasma and serum CA19-9 were evaluated in periampullary carcinoma patients (n = 109) undergoing Whipple's pancreaticoduodenectomy and in healthy volunteers (n = 92). Tumour tissues were subjected to staging and subtyping prior to determining differentially expressed miRNAs in them by quantitative real-time polymerase chain reaction (qRT-PCR). Statistical analysis involved correlation, receiver operating characteristic, logistic regression, survival analyses and Cox-proportional regression. Of the three differentially expressed circulating miRNA, miRNA192 was significantly increased both in circulation and in tumour tissue and correlated with tumour stage and aggressiveness (r = 0.96, P < 0.0001). Area under the curve of circulating miRNA192 + CA19-9 combination was 0.877 (95% confidence interval, 0.72 to 0.96) for stage III and 0.92 (95% confidence interval, 0.77 to 0.88) for tumour aggressiveness. The combination was associated with poor survival (median: 22 months, P = 0.0008) in stage III patients. Cox-proportional regression analysis revealed prognostic importance of combination of circulating miR192 and CA19-9 (HR = 1.005, P = 0.0001) in periampullary carcinoma. In conclusion, combination of circulating miRNA192 with serum CA19-9 is a better prognostic biomarker than CA19-9 alone.

Expression profiling of maternal plasma and placenta microRNAs in preeclamptic pregnancies by microarray technology

Preeclampsia (PE) is one of the leading causes of maternal and fetal morbidity and mortality, occurring usually in the second half of pregnancy and affecting approximately 5–8% of pregnancies in the world. miRNAs play critical role in the regulation of placental development processes. We aimed to determine specific novel miRNAs for early diagnosis of preeclampsia which is one of the most dangerous pregnancy diseases. In this study 72 samples, maternal age 22 ≤ and ≤36, have been analyzed; maternal plasma and placental miRNAs were isolated from 18 severe preeclampsia (sPE) patients and 18 controls, respectively. Profiling of human miRNAs (1368 probe) was performed in samples with Agilent v16 microarrays for detection of the differences in miRNA expression between two groups. The results were validated by using TaqMan RT-qPCR method. The analysis indicated that 406 of these miRNAs in all placentas and 42 of these miRNAs in all maternal plasma were expressed. The relative expression analysis has shown that 12 miRNAs (p < 0.05 and >2-fold) in maternal plasma were differentially expressed in PE and control group. However, five miRNAs were validated by qRT-PCR. Once validated miRNAs have been searched in databases for their target genes and function, it has been shown that there are some preeclampsia related pathways as a target such as angiogenesis, cardiovascular, hypertension, placental abruption and preeclampsia disorders. Differentially expressed and validated plasma miRNAs might be used as notable biomarkers for non-invasive early diagnosis of preeclampsia and treatment of disease.

Identification of miRNAs as non-invasive biomarkers for early diagnosis of lung cancers.

Current clinical diagnostic methods lack the specificity in detecting lung cancer patients. The issue is particularly critical for stage I and II patients. Considerable evidence showed microRNA plays a very important role in lung carcinogenesis. Here, we identified a panel of 41 miRNAs significantly elevated in patients with lung cancer, of which eight miRNAs were further validated in an independent sample cohort. Classification analysis using the panel of eight miRNAs generated a discriminatory power of 93.3% sensitivity and 93.8% specificity in separating non-small-cell lung cancer (NSCLC) patients from normal controls, indicating the miRNAs have a potential clinical utility in discriminating NSCLC. Interestingly, miR-1244 was found significantly elevated in the serum samples of lung cancer patients, and the test characteristics of the single miRNA were area under the curve (AUC) of 0.832 in NSCLC vs healthy controls, and 0.861 in NSCLC vs patients with unidentified pulmonary nodules. This is the first study showing serum miR-1244 could be a biomarker to screen lung cancer patients from the high-risk population.

How strong is the association between IPF and lung cancer? An answer from airway's DNA.

Idiopathic pulmonary fibrosis is a chronic progressive disease of lung interstitium of unknown etiology with poor prognosis. In patients with IPF, the incidence of lung cancer is much higher than that in the general population. The identification of noninvasive biomarkers for early diagnosis of IPF is of great relevance in consideration of the management of these patients. Among the noninvasive omic markers, an increasing interest has been directed toward the study of genetic alterations of microsatellites (MAs) in exhaled breath condensate (EBC). The aim of this preliminary study was to investigate the MAs, located in chromosomal regions 8p21.3-q11.1 and 17q11.2-q21, that harbor tumor suppressor genes, in EBC and in the paired whole blood (WB) of IPF patients. Eleven IPF patients were compared with 10 healthy control subjects. All subjects underwent collection of the EBC and WB. The EBC was collected using a condenser. Four microsatellite markers (THRA1, D17S579, D17S250 and D8S137) were used for the analysis of MAs. The EBC-DNA and WB-DNA were amplified by PCR; PCR products were analyzed using the ABI Prism 310 DNA. Microsatellite alterations were found in 58.82% of EBC-DNA and 12.50% of WB-DNA in patients with IPF (p<0.01). None of the healthy subjects exhibited MAs in the studied markers. Our findings suggest that these genetic alterations, studied in EBC, may play an important role in the complex genetic basis of IPF. Since these MAs are frequently detected in cancer, they might explain the higher relative risk of tumorigenesis in this disease.

MicroRNAs as noninvasive biomarkers in bladder cancer detection: a diagnostic meta-analysis based on qRT-PCR data.

As the diagnostic significance of microRNAs (miRNAs) in the detection of bladder cancer is controversial, we aimed to perform a meta-analysis to comprehensively assess the diagnostic value of miRNAs in blood and urine for detecting bladder cancer. A systematic literature search of public databases was conducted to obtain qualified studies. Sensitivity was utilized to plot the summary receiver operator characteristic (SROC) curve against specificity and the area under the SROC curve (AUC) was generated to evaluate the pooled diagnostic efficiency. Subgroup analyses and meta-regression were applied to investigate the underlying sources of heterogeneity. The STATA 12.0 software was used to perform all statistic analyses. A total of 58 studies from 22 articles comprising 4,558 bladder cancer patients and 4,456 controls were included in our meta-analysis. MiRNAs in blood and urine manifested relatively good diagnostic efficiency in detecting bladder cancer, with a sensitivity of 0.74, a specificity of 0.78, and an AUC of 0.83. Multiple-miRNA assays were more accurate than single-miRNA ones in bladder cancer diagnosis. Blood-based miRNA assays displayed better diagnostic performance than urine-based ones. In addition, miRNAs showed reduced diagnostic value in bladder cancer among Caucasians compared with Asians. MiRNAs in blood and urine, especially the combination of multiple miRNAs, may serve as hopeful noninvasive biomarkers for early diagnosis of bladder cancer. Further extensive prospective research is needed to verify their clinical significance in bladder cancer diagnosis.

Stage-wise comparison of anti-endometrial-antibodies against peptides of SLP2, TMOD3 and TPM3 in diagnosis of endometriosis

The lack of non-invasive diagnostic test for early diagnosis of endometriosis results in 8-11 years delay in diagnosis leads to deterioration of quality of life. Previously, we identified anti-endometrial-antibodies against peptides of Stomatin like protein 2 (SLP2), Tropomodulin 3 (TMOD3) and Tropomyosin 3 (TPM3), and further proposed their utility as non-invasive biomarkers for early diagnosis of endometriosis.The aim of the present study was to investigate whether the levels of biomarkers vary with disease progression and to compare the sensitivity, specificity and diagnostic accuracy of the biomarkers in diagnosis of endometriosis in early stages (Stage I-II) versus advanced stages (Stage III-IV). Multi-centre, cross sectional study Women with endometriosis (Stage I-II n=133, Stage III-IV n=133)and healthy controls (n=104) were screenedfor eleven novel autoimmune markers (anti-endometrial-antibodies of SLP2a, SLP2b, SLP2c, TMOD3a, TMOD3b, TMOD3c, TMOD3d, TPM3a, TPM3b, TPM3c and TPM3d) using the peptide ELISA.The statistical analysis was performed using STATA software (version 8.2, Texas, USA). The mean serum levels of anti-endometrial-antibodies of SLP2a, SLP2b, SLP2c, TMOD3a, TMOD3b, TMOD3c, TMOD3d, TPM3a, TPM3b, TPM3c and TPM3d) in early stages (Stage I-II) were significantly higher than that of advanced stages (Stage III-IV) of endometriosis (p<0.05). The sensitivity and diagnostic accuracy of all these biomarkers were higher in detection of Stage I-II endometriosis than in the detection of Stage III-IV endometriosis at an acceptable specificity of ≥60%. We observed that the levels of serum anti-endometrial antibodies to SLP2, TMOD3 and TPM3 did not appear to elevate further in advanced stages (Stage III-IV) of endometriosis.

Abstract LB-247: Circulating proteolytic products of tumor-resident enzyme as potential biomarkers for early detection of breast cancer

Abstract In theory, any or all of tumor-secreted proteins can serve as cancer biomarkers. However, the reality is challenging to monitor because of the large degree of fluctuation in abundance and localization of these tumor-secreted proteins, especially in the early stage of tumor development and/or metastasis. As such, it seems feasible that we might take advantage of the fact that secreted proteases/peptidases in the tumor microenvironment generate proteolytic products, also referred to as “circulating peptides”, which are detectable in bloodstream and provide ample information about the body, “coded” in the patterns and quantity of these peptides. Herein we clearly link the catalytic activity of Carboxypeptidase N (CPN) to its proteolytic products during breast tumor progression in mouse model and clinical samples. CPN plays important roles in regulating vasoactive peptide hormones, growth factors, and cytokines by specifically cleaving their C-terminal basic residues. Circulating fragment profiling, by an approach combining nanopore fractionation and mass spectrometry, revealed the nature and extent of cleavage by CPN. These results correlated with the level of CPN-catalyzed peptides in blood taken from the tumor-bearing mice, healthy women and breast cancer patients. We showed that generation of C3f_R1310-L1319 specifically correlated with the CPN expression level. In both mouse and clinical patient samples, the amount of CPN was increased in tumor tissues compared to that seen in normal breast tissue, while its counterpart in blood remained constant. The amount of 6 CPN-catalyzed peptides predominantly increased in sera taken from both the mice at 2 weeks after orthotropic implantation and the patients’ plasma as early as the first pathologic stage of breast cancer. In conclusion, the circulating level of the selected 6 CPN-catalyzed proteolytic products reflects the extent of this enzyme's activity in tumors, and our results clearly indicate their strong potential as biomarkers for non-invasive early diagnosis of breast cancer. Citation Format: Ye Hu. Circulating proteolytic products of tumor-resident enzyme as potential biomarkers for early detection of breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-247. doi:10.1158/1538-7445.AM2014-LB-247