Serum and Urinary Biomarkers for Early Detection of Anisakis Simplex Infection in a Rodent Model: A Non-Invasive Approach to Zoonotic Nematode Diagnosis

Hepatocellular carcinoma (HCC) is previously considered a rare type of cancer in the Western world, but shows an annual increase of 3.5% since 1992 and is currently ranked second only to pancreatic cancer for cancer-related deaths in the United States due to its lack of early detection markers and poor prognosis. HCC is particularly difficult to treat because its development impairs liver normal drug metabolic function leading to chemoresistant. Although chronic infections with hepatitis B or C virus are the major risk factors for HCC worldwide, liver metabolic diseases associated with obesity such as non-alcoholic fatty liver diseases, diabetes mellitus and iron-storage diseases, account for the majority of HCC cases in the U.S. The rapid increase in obesity and obesity-associated metabolic disease in recent years suggests that the incidence of metabolic dysfunction-induced HCC is likely continually to increase in the States in the future. However, most existing HCC animal models develop tumors either following the treatment of carcinogens or ablation of a key liver tumor suppression pathway, which do not follow the same pathophysiological mechanisms of tumor initiation and progression as metabolic disorder-induced HCC in humans. Hence, there is a critical need to establish animal models that develop spontaneous HCC in response to chronic metabolic stress to study the mechanism of metabolic dysfunction-induced HCC, identify early detection markers associated with these types of HCC, and investigate the role of liver metabolic disruption in anticancer treatment. Most physiological processes including nutrient intake, storage and metabolism follows a circadian rhythm in mammals since our homeostatic systems are shaped by the evolutionary adaptation to daily light/dark changes in the environment. Disruption of circadian homeostasis leads to a coupled increase in the risk of obesity, liver metabolic syndromes and cancers in both night-shift human workers and animal models. We have previously reported that circadian gene-mutant mouse models show a significant increased risk to cancer. Our recent studies have revealed that disrupting circadian rhythm in wild-type mice by chronic jet-lag following a schedule that mimics the night-shift working schedules in humans leads to a progressive deregulation of multiple plasma and hepatic metabolic parameters in the serum and induction of multiple liver metabolic syndromes including hepatosteatosis as well as liver inflammation and uncontrolled hepatocytes and bile duct proliferation prior to HCC onset. Genome-wide array analyses have led to identify a time-dependent deregulation of all known key pathways controlling metabolism, energy storage, redox levels, cell proliferation, inflammatory response and tumor suppression in the liver of mice lacking circadian homeostasis. Our studies have established an excellent mouse model to study the role of metabolic dysfunction in spontaneous HCC induction. Further studies will lead to elucidate the mechanism of HCC induction by chronic metabolic disruption and identify novel serum and hepatic biomarkers for HCC early detection and treatment. Citation Format: Nicole M. Kettner, David D. Moore, Loning Fu. Studying circadian disruption as a novel risk factor of hepatocellular carcinoma using mouse models. [abstract]. In: Proceedings of the AACR Special Conference: The Translational Impact of Model Organisms in Cancer; Nov 5-8, 2013; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2014;12(11 Suppl):Abstract nr A32.

Melanoma is one of the most lethal and malignant cancers and its incidence is increasing worldwide, and Japan is not an exception. Although there are numerous therapeutic options for melanoma, the prognosis is still poor once it has metastasized. The main concern after removal of a primary melanoma is whether it has metastasized, and early detection of metastatic melanoma would be effective in improving the prognosis of patients. Thus, it is very important to identify reliable methods to detect metastases as early as possible. Although many prognostic biomarkers (mainly for metastases) of melanoma have been reported, there are very few effective for an early diagnosis. Serum and urinary biomarkers for melanoma diagnosis have especially received great interest because of the relative ease of sample collection and handling. Several serum and urinary biomarkers appear to have significant potential both as prognostic indicators and as targets for future therapeutic methods, but still there are no efficient serum and urinary biomarkers for early detection, accurate diagnosis and prognosis, efficient monitoring of the disease and reliable prediction of survival and recurrence. Levels of 5-S-cysteinyldopa (5SCD) in the serum or urine as biomarkers of melanoma have been found to be significantly elevated earlier and to reflect melanoma progression better than physical examinations, laboratory tests and imaging techniques, such as scintigraphy and echography. With recent developments in the treatment of melanoma, studies reporting combinations of 5SCD levels and new applications for the treatment of melanoma are gradually increasing. This review summarizes the usefulness of 5SCD, the most widely used and well-known melanoma marker in the serum and urine, compares 5SCD and other useful markers, and finally its application to other fields.

DETECT: Development of Technologies for Early HCC Detection

The devastatingly high level of mortality associated with pancreatic ductal adenocarcinoma (PDAC), the fourth leading cause of cancer death in the United States, has remained virtually unchanged over the past 50 years due to an inability to effectively detect and treat the disease at an early stage. Currently used imaging techniques including CT and MRI have demonstrated an inability to distinguish benign from malignant pancreatic lesions resulting in both a low sensitivity for pancreatic cancer and a high degree of unnecessary referral for highly invasive endoscopic ultrasound (EUS). A biomarker-based test that could effectively rule out the possibility of malignancy in patients presenting with an abnormal finding by CT or MRI would greatly reduce the number of unnecessary EUS procedure and the associated risk of complication as well as lead to an optimization of resources for more effective early detection of the disease. Serum biomarkers such as CA 19-9 have been under investigation in pancreatic cancer for a number of years. Recently, the use of urine in biomarker studies has garnered considerable interest based on its ease of sampling, high stability, and relative simplicity as a testing matrix. In order to evaluate the use of urine and serum biomarkers in pancreatic cancer screening, we analyzed 77 cancer-associated biomarkers in urine obtained from 55 patients diagnosed with PDAC and 44 patients diagnosed with benign pancreatic conditions. Twenty of the most relevant biomarkers were also tested in sera obtained from the same patient cohort. Forty of the 77 urine biomarkers as well as nine of the twenty serum biomarkers were significantly altered between the benign and malignant groups. Of the twenty biomarkers measured in both urine and serum, seven were more significantly altered in urine, while seven others were more significantly altered in serum. A multivariate analysis of the results identified a four-biomarker panel comprised of both serum and urine biomarkers which could discriminate PDAC from benign disease with optimal levels of sensitivity and specificity. Our results indicate a promising role for the combined use of urine and serum biomarkers in the development of improved screening strategies for pancreatic cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3181. doi:10.1158/1538-7445.AM2011-3181

Biosensor for electrochemical detection of biomarkers of osteoarthritis

Ovarian cancer (OC) is one of the leading causes of gynaecological cancer mortality in women worldwide. If detected at an early stage (I, II), OC has a 90% 5-year survival rate; nevertheless, symptoms are often hidden, leading to late-stage (III, IV) diagnosis and a poor prognosis. The current diagnostic procedures, such as a pelvic exam, transvaginal ultrasound, CA-125 blood tests, serum HE4 tests and multivariate index assays (MIA), are insufficient. Sadly, surgery is frequently required to confirm a positive diagnosis. Therefore, there has been an increased interest in different biomarkers using a non-invasive test as a tool for the earlier diagnosis of OC to resolve the need for precise and non-invasive diagnostic methods. This review article aims to investigate how biomarkers influence early OC detection and to emphasise the role of using a combination of serum biomarkers panel rather than a single biomarker. In addition, this review provides insights into the current serum biomarkers, urine biomarkers and other emerging biomarkers in the early detection of OC for better specificity and sensitivity and to improve the overall survival (OS) rate.

Blood and urine biomarkers associated with long-term respiratory dysfunction following neonatal hyperoxia exposure: Implications for prematurity and risk of SIDS

The separation of biomarkers from blood is straightforward in most molecular biology laboratories. However, separation in resource-limited settings, allowing for the successful removal of biomarkers for diagnostic applications, is not always possible. The situation is further complicated by the need to separate hydrophobic signatures such as lipids from blood. Herein, we present a microfluidic device capable of centrifugal separation of serum from blood at the point of need with a system that is compatible with biomarkers that are both hydrophilic and hydrophobic. The cross-flow filtration device separates serum from blood as efficiently as traditional methods and retains amphiphilic biomarkers in serum for detection.

To provide a comprehensive review of the available biomarkers for the detection and active surveillance of prostate cancer and simplify decision-making while choosing between them. The limitations of PSA and mpMRI and the invasive nature of prostate biopsy has led to a constant search for serum and urinary biomarkers for both the detection and monitoring during active surveillance of prostate cancer. 4K, PHI and PCA3 have been validated in prospective clinical trials for initial detection of prostate cancer and recent evidence points to potential differentiation between indolent and aggressive cancer. However, the usage in monitoring tumor dynamics is debatable because of lack of conclusive evidence. The answer to the existing problems lies in high-quality studies to establish definitive evidence and also to help choose between the plethora of biomarkers available today. Despite the advancements in innovation and usage of biomarkers in prostate cancer, there exists tremendous potential in improving them to fulfil the unmet need that exists today. Studies to establish conclusive evidence and integration with imaging can tremendously aid diagnosis and monitoring.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive cancer with a poor prognosis, primarily due to a late diagnosis. Recent studies have focused on identifying non-invasive biomarkers for early detection, with microRNAs (miRNAs) emerging as promising candidates. This systematic review aims to evaluate the potential of circulating miRNAs as biomarkers for the early detection of PDAC, analyzing their diagnostic accuracy, specificity, and sensitivity. Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a comprehensive search across PubMed, Embase, and the Cochrane Library was conducted. Studies published from January 2013 to October 2023 focusing on miRNA biomarkers for early PDAC detection were included. Data synthesis was performed through a narrative approach due to the heterogeneity of the studies. Nine studies met the inclusion criteria. Key findings include the elevated levels of specific miRNAs, such as miR-18a, miR-106a, and miR-25, in early-stage PDAC patients compared to controls. The integration of miRNA profiles with traditional biomarkers like CA19-9 showed improved diagnostic performance. However, challenges in the standardization of miRNA evaluation methodologies were noted. Circulating miRNAs demonstrate significant potential as non-invasive biomarkers for early PDAC detection. Despite promising results, further research and standardization are necessary for clinical application.

China has 50% of the worldwide hepatocellular carcinoma (HCC) cases, and the HBV-related cases accounts for approximately 85%. Over the past few decades, although a series of standardized management methods for HCC has been implemented in China, most HCC patient in China still suffered from advanced-stage disease, in consequence, reducing the opportunity of curable treatment that can be offered to achieve long-term disease-free survival for HCC patient. Accordingly, strategies including screening and diagnose HCC at an earlier stage are urgently needed in China. In this study, the current status, challenges, and prospects of early detection of HCC in China have been analyzed. The result indicated the need for using multi serum biomarkers for early HCC detection. During the past ten years, the research on the clinical usefulness of novel serum biomarkers of des-γ-carboxy-prothrombin (DCP), Dickkopf-1 (DKK1) and Midkine (MDK) in early HCC detection for Chinese patients found that the novel serum biomarker can complete the measurement of α-fetoprotein (AFP) in the diagnosis process of HCC, particularly for the patient with negative AFP with/or at an early stage. More large-scale, multi-center studies are expected to be performed in China to provide further evidence, and using novel and reliable serum biomarkers to complement AFP as a new trend is expected to be extensively used in clinical practice to facilitate early detection for those patients with HCC in China.

Serum and tissue biomarkers have begun to play an increasingly important role in the detection and management of many cancers of hormone-sensitive tissues. Specifically, the introduction of serum PSA measurements into clinical practice has dramatically altered detection and treatment of prostate cancer and serum tumor markers play a critical role in the management of testicular cancer. Serum biomarkers are used for ovarian and pancreatic cancers, but their usefulness is limited by poor specificity. Tissue biomarkers are used to help guide breast cancer treatment but are not widely used in other cancers. Even the "best" biomarkers such as PSA have substantial limitations. The discovery of new biomarkers for both early detection and prognosis of cancer is critical to the hope of better clinical outcomes. Recently there has been an expanding understanding of the underlying molecular etiology of cancer and molecular targeted therapies for some particularly aggressive cancers such as renal cell carcinoma have been developed. Better understanding of the molecular etiology of cancer and identification of additional therapeutic targets remain important research goals. Currently, there are very few patient-tailored therapies and there is a great need to better understand the molecular alterations associated with cancer and to use this information to design need cancer therapies and prevention strategies. Advances in proteomic technologies have created tremendous opportunities for biomarker discovery and biological studies of cancer. The potential that proteomics will impact clinical practice is currently greater than ever, but there main several obstacles in making this a reality. A major hurdle to overcome continues to be the proper acquisition of patient tissues and body fluids for investigation and clinical diagnostics. Each cancer has specific issues in this regard and it is incumbent upon investigators and collaborating clinicians to understand the various nuances of tissue and biofluid procurement. This chapter not only reviews the clinical need and potential impact of proteomic studies of hormone-sensitive cancers, but details specific technologies and discusses the issues surrounding tissue/biofluid procurement.

Background:Diffuse-type gastric cancer (DGC) exhibits rapid disease progression and a poor prognosis. There are no effective serum biomarkers for early detection of DGC. We have established an E-cadherin/p53 double conditional knockout (DCKO) mouse line that recapitulates human DGC morphologically and molecularly. In this study we tried to identify circulating microRNAs (miRNAs) as non-invasive biomarkers for DGC diagnosis using DCKO mice.Methods:We performed miRNA microarray and quantitative reverse transcription–PCR analyses of tissue and serum samples from DCKO mice with DGC and age-matched littermate controls.Results:Comparative analyses showed that mouse and human primary gastric cancers have similar miRNA expression patterns. Next, we selected some candidate miRNAs highly expressed in sera and cancer tissues of DCKO mice for further evaluation. TaqMan quantitative RT–PCR analyses indicated that four of them, miR-103, miR-107, miR-194 and miR-210, were significantly upregulated in sera of both early and advanced-stage DGC-bearing mice compared with in corresponding controls. Receiver-operating characteristic curve analyses demonstrated that these four miRNAs can discriminate DGC-positive cases from normal ones with high sensitivity and specificity.Conclusion:These observations suggest that this mouse model of DGC is useful for identifying serum biomarkers, and we found circulating miRNAs that can accurately detect DGC at an early stage.

Simple SummaryDetecting cancer early significantly increases the chances of successful (surgical) treatment. Pancreatic cancer is one of the deadliest cancer forms, since it is usually discovered at a late and already spread stage. Finding biomarkers showing pancreatic cancer at an early stage is a possible approach to early detection and improved treatment. The aim of our study was to assess the potential of tissue polypeptide specific antigen (TPS) as a biomarker for early pancreatic cancer detection. We studied TPS levels in blood plasma samples from a population-based biobank in Västerbotten, Sweden that were collected before individuals were diagnosed with pancreatic cancer. Although TPS levels are raised at diagnosis, this occurs late, and thus TPS does not seem to hold promise as an early detection marker for pancreatic cancer.Early detection of pancreatic ductal adenocarcinoma (PDAC) is challenging, and late diagnosis partly explains the low 5-year survival. Novel and sensitive biomarkers are needed to enable early PDAC detection and improve patient outcomes. Tissue polypeptide specific antigen (TPS) has been studied as a biomarker in PDAC diagnostics, and it has previously been shown to reflect clinical status better than the ‘golden standard’ biomarker carbohydrate antigen 19-9 (CA 19-9) that is most widely used in the clinical setting. In this cross-sectional case-control study using pre-diagnostic plasma samples, we aim to evaluate the potential of TPS as a biomarker for early PDAC detection. Furthermore, in a subset of individuals with multiple samples available at different time points before diagnosis, a longitudinal analysis was used. We assessed plasma TPS levels using enzyme-linked immunosorbent assay (ELISA) in 267 pre-diagnostic PDAC plasma samples taken up to 18.8 years before clinical PDAC diagnosis and in 320 matched healthy controls. TPS levels were also assessed in 25 samples at PDAC diagnosis. Circulating TPS levels were low both in pre-diagnostic samples of future PDAC patients and in healthy controls, whereas TPS levels at PDAC diagnosis were significantly increased (odds ratio 1.03; 95% confidence interval: 1.01–1.05) in a logistic regression model adjusted for age. In conclusion, TPS levels increase late in PDAC progression and hold no potential as a biomarker for early detection.

Pancreatic cancer is one of the most fatal human cancers, with an overall 5-year survival rate of 10.8%. Early detection is critical for improving pancreatic cancer prognosis, but biomarkers for early detection are lacking. We conducted a two-stage study to identify circulating miRNAs as biomarkers for pancreatic cancer early detection using pre-diagnostic plasma samples, collected within 5 years prior to cancer diagnosis, from case-control studies nested in five prospective studies. The discovery stage included 185 case-control pairs from the Prostate, Lung, Colorectal and Ovarian Cancer (PLCO) Screening Trial. Replication stage samples comprised 277 case-control pairs from diverse cohorts: Shanghai Women’s and Men’s Health Studies, Southern Community Cohort Study (SCCS), and Multiethnic Cohort. Controls were individually matched on age at enrollment, sex, recruitment site (SCCS), race/ethnicity, and date of blood draw in each cohort. Cell-free small RNAs were extracted from plasma samples and miRNAs were measured by the NanoString nCounter Analysis System using the Human v3 miRNA Expression panel (a total of 798 miRNAs). Normalized miRNAs were categorized by decile. For miRNAs that have ≥10% samples with an undetectable level (0), the non-zero level was categorized by approximately 10% increment. Associations of circulating miRNAs with pancreatic cancer risk, measured in odds ratios (ORs) and 95% confidence intervals (CIs) per decile change, were calculated using conditional logistic regression analyses in discovery and replication studies, separately within each cohort, and results meta-analyzed. We identified three miRNAs, hsa-miR-199a-3p+/hsa-miR-199b-3p, hsa-miR-191-5p, hsa-miR-767-5p, being consistently associated with pancreatic risk in both discovery and replication sets, with combined ORs (95% CIs) of 0.89 (0.84-0.95), 0.90 (0.85-0.95), and 1.08 (1.02-1.13), and P of 9.09E-05, 6.95E-05 and 4.03E-03, respectively. Adjustment for age, BMI, smoking, diabetes and family history of pancreatic cancer did not change the associations. Stratified analyses by age at diagnosis found five additional replicated miRNAs: hsa-miR-640, hsa-miR-1299, hsa-miR-22-3p, hsa-miR-874-5p, and hsa-miR-449b-5p among those 65 years or older, with combined ORs (95% CIs) of 1.33 (1.16-1.52), 1.28 (1.12-1.46), 0.76 (0.65-0.89), 1.25 (1.09-1.43), and 1.22 (1.07-1.39), and P-value ranging from 4.75E-05 to 0.003. These results suggest that circulating miRNA biomarkers may be useful in identifying individuals with high risk of developing pancreatic cancer for close surveillance and/or a screening test. Citation Format: Cong Wang, Hui Cai, Qiuyin Cai, Jie Wu, Rachael Stolzenberg-Solomon, Clair Zhu, Yu-Tang Gao, Jordan Berlin, Fei Ye, Wei Zheng, Veronica W. Setiawan, Xiao-Ou Shu. Circulating miRNA as biomarkers for pancreatic cancer early detection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB109.