Detection Of Early Stage Pancreatic Cancer Research Articles

Abstract 5960: Blood-based extracellular vesicle biomarker test for detection of early-stage pancreatic cancer

Abstract Pancreatic cancer is an almost uniformly lethal malignancy, in large part because is it rarely detected in its early, curable stages. We recently conducted a study to evaluate the use of proteins associated with extracellular vesicles (EVs) isolated using an alternating current electrokinetic (ACE) system for the early detection of pancreatic ductal adenocarcinoma. Because earlier detection of pancreatic cancer greatly improves outcomes, we sought to devise a blood-based test with high specificity for detecting early-stage PDAC based on identification of EV-bound biomarkers. EVs were purified from control or stage I-II pancreatic ductal adenocarcinoma (PDAC) patient plasma using an ACE microelectrode array. Following collection of EVs onto the electrodes, contaminating plasma elements were washed away, and the electric field was turned off to release EVs into buffer. The isolated EV/exosome samples were then tested using bead-based multiplex immunoassay kits to determine relative concentrations of biomarkers of interest. Sample cohorts were divided into a training set (controls, N=146; PDAC cases, N=33) to develop a logistic regression-based classification model, and a validation set (controls, N=139; PDAC cases, N=35) to generate a probability-based score for predicting the presence of PDAC. For the training set, the algorithm identified six protein biomarkers associated with the presence of stages I-II cancer in PDAC patient plasma, with a receiver-operating characteristic (AUC) of 0.997 (95% CI: 0.978-1.000) compared to control plasma samples. For the validation set, the algorithm had a sensitivity of 82.9% (95% CI: 67.3% - 91.9%) at a specificity of 99.3% (95% CI: 96.0% - 99.9%), generating an AUC = 0.978 (95% CI=0.954-0.995). The exosomal PDAC assay shows feasibility for developing minimally invasive liquid biopsies that allow low-risk, routine screening of high-risk patients. Those patients identified by the test would be able to undergo additional testing to determine if there is PDAC present. The additional screening and testing of these patients may increase the likelihood of curative treatment. Further work in large datasets is warranted. Citation Format: Juan P. Hinestrosa, Jean M. Lewis, Heath I. Balcer, Razelle Kurzrock, Scott Lippman, Rajaram Krishnan. Blood-based extracellular vesicle biomarker test for detection of early-stage pancreatic cancer [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 5960.

Plasma miRNA Biomarkers in Limited Volume Samples for Detection of Early-stage Pancreatic Cancer.

Early detection of pancreatic ductal adenocarcinoma (PDAC) is key to improving patient outcomes; however, PDAC is usually diagnosed late. Therefore, blood-based minimally invasive biomarker assays for limited volume clinical samples are urgently needed. A novel miRNA profiling platform (Abcam Fireplex-Oncology Panel) was used to investigate the feasibility of developing early detection miRNA biomarkers with 20 μL plasma from a training set (58 stage II PDAC cases and 30 controls) and two validation sets (34 stage II PDAC cases and 25 controls; 44 stage II PDAC cases and 18 controls). miR-34a-5p [AUC = 0.77; 95% confidence interval (CI), 0.66-0.87], miR-130a-3p (AUC = 0.74; 95% CI, 0.63-0.84), and miR-222-3p (AUC = 0.70; 95% CI, 0.58-0.81) were identified as significantly differentially abundant in plasma from stage II PDAC versus controls. Although none of the miRNAs individually outperformed the currently used serologic biomarker for PDAC, carbohydrate antigen 19-9 (CA19-9), combining the miRNAs with CA 19-9 improved AUCs from 0.89 (95% CI, 0.81-0.95) for CA 19-9 alone to 0.92 (95% CI, 0.86-0.97), 0.94 (95% CI, 0.89-0.98), and 0.92 (95% CI, 0.87-0.97), respectively. Gene set enrichment analyses of transcripts correlated with high and low expression of the three miRNAs in The Cancer Genome Atlas PDAC sample set. These miRNA biomarkers, assayed in limited volume plasma together with CA19-9, discriminate stage II PDAC from controls with good sensitivity and specificity. Unbiased profiling of larger cohorts should help develop an informative early detection biomarker assay for diagnostic settings. PREVENTION RELEVANCE: Development of minimally invasive biomarker assays for detection of premalignant disease and early-stage pancreatic cancer is key to improving patient survival. This study describes a limited volume plasma miRNA biomarker assay that can detect early-stage resectable pancreatic cancer in clinical samples necessary for effective prevention and clinical intervention.

Identification of potential common molecular factors of pancreatic cancer and diabetes mellitus using microarray data analysis combined with bioinformatics techniques and experimental validation

Background and Aim: Detection of early-stage pancreatic cancer (PaC) patients can decrease the mortality rate through surgical resection thanks to a screening approach of high-risk and asymptomatic individuals. Up to 80% of PaC patients are either hyperglycemic or diabetic at the time of diagnosis. Diabetes mellitus (DM) identification as an early manifestation of PaC may lead to detection of this malignancy at early and presymptomatic stages. Therefore, the current study aims to identify common molecular factors between DM and PaC to shed light on a potential screening for DM as a diagnostic approach for PaC. Methods: The study was divided into three steps including dataset selection and identification of key genes, quantitative real-time polymerase chain reaction (qRT-PCR) to validate these genes, and enrichment analysis of their target genes. Results: Using GEO2R analysis, conducted on microarray dataset (GSE15932), SPI1 and Yin Yang 1 (YY1) genes were found to be expressed in peripheral blood mononuclear cells of PaC, DM, and PaC + DM patients. Moreover, qRT-PCR results in validation of microarray data showed a significant increment of these two genes among all patient groups. Furthermore, enrichment analyses of SPI1 and YY1 target genes revealed that they are mainly expressed in hematopoietic cells and associated with immune responses as well as immune signaling pathways. Discussion: We speculate that this study on SPI1 and YY1 genes and their targets can result in a successful strategy to investigate diabetes as a screening step for better management of PaC screening using standard serologic tests.

A simple MWCNTs@paper biosensor for CA19-9 detection and its long-term preservation by vacuum freeze drying

This paper introduces a cheap simple MWCNTs@paper biosensor for the detection of CA19-9, which is a biomarker of pancreatic cancer. By adding the CA19-9 antibody to the surface of MWCNTs which are deposited on the microporous filter paper, the correlation between the concentration of CA19-9 and resistance of biosensor element was linear due to the site-specific binding of antigen and antibody. The detection range is wide (0 U/mL–at least 1000 U/mL), and even in the low concentration of CA19-9, the linearity remains satisfying. Based on this property, it could be used for the detection of early-stage pancreatic cancer. Besides, this research originally introduces a vacuum freeze-drying method for the long-term preservation of biosensor, prolonging its storage time from 3 h to at least 7 days, which signifcantly promoted its value in practical application. One piece of the MWCNTs@paper biosensor only cost $2 (about 30 times cheaper than ELISA) approximately, and the detection speed is satisfying (2 h, 12 times faster than ELISA), which will possibly increase its opportunity of mass production and clinical practice.

Abstract 1040: Characterization of extracellular vesicles in pancreatic cancer: Changing the way we think about tumorigenesis

Abstract Introduction: Pancreatic ductal adenocarcinoma (PDAC) is projected to become the second leading cause of cancer-related deaths by 2030. Patients diagnosed with PDAC have a 5-year survival rate ~9%. Detection of pre-neoplastic lesions has the potential to improve treatment and overall survival in pancreatic cancer (PC) by 4-5 fold. However, there is currently no validated screening test for the non-invasive and accurate detection of early stage PC. Extracellular vesicles (EVs) arise from invagination of, and secretion from, cells of all tissue types. Their role in mediating PC tumorigenesis however, is just beginning to be studied. We have found that EVs derived from PC cells (cancer-derived EVs, CDEVs), can impart significant phenotypic and metabolic transformational effects on normal human pancreas cells (hTERT-HPNE and H6c7), tantamount to an epithelial-to-mesenchymal transition (EMT). The goal of this study was to characterize the biochemical cargo of EVs toward understanding their role in PC tumorigenesis. Results: We isolated CDEVs from various established and PDX PC cell lines. After validating enrichment of EV fractions with quantitative ELISA, immunoblot and transmission electron microscopy (TEM), we added CDEVs to culture media of normal cells. We found that CDEVs induce a myriad of changes in normal cells, including significant morphological changes, increased proliferation and an uncharacteristic invasive capability. CDEVs also caused a bioenergetic switch in normal cells, from a quiescent, aerobic profile to a highly energetic and glycolytic profile. Comparative analyses of CDEVs and normal-derived EVs (NDEVs) using 1H- and 13C-NMR showed significantly different biochemical fingerprints. Interestingly, we found that NDEVs contained more amino acids than CDEVs, while CDEVs contained elevated amounts of urea and lactic acid. We next designed a 13C-isotope labeling flux experiment to trace the fate of carbon metabolized from 13C-glucose fed to cells in culture. Using UHPLC/Q-ToF-MS, we identified several differentially labeled isotopes in CDEVs, particularly surrounding lipid biosynthesis and folate. Conclusions: Our results indicate that CDEVs confer enormous transformational properties to normal human pancreas cells in vitro. We hypothesize that EVs could impart tumorigenic properties to normal cells in vivo and this influence could unveil novel mechanisms underpinning cancer onset and progression. We show that there are robust biochemical differences between CDEVs and NDEVs. Biochemical characterization of EV cargo can uncover abrogated pathways in which signaling is mediated by EVs. These signals may be detectable before progression of PC to PDAC, leading to the development of assays for earlier diagnosis in patients. Citation Format: Charles P. Hinzman, Michael Girgis, Yaoxiang Li, Meth Jayatilake, Meena Rajagopal, Partha P. Banerjee, Amrita K. Cheema. Characterization of extracellular vesicles in pancreatic cancer: Changing the way we think about tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1040.

Plasma biomarker for detection of early-stage pancreatic cancer and risk factors for pancreatic malignancy using antibodies for apolipoprotein-A2 isoforms.

4106Background: We have previously revealed that circulating levels of apolipoprotein A2 (apoA2) isoforms apoA2-ATQ/AT (C-terminal truncations of apoA2 homo-dimer) are significantly decreased in ea...

Abstract 4580: Detection of early-stage pancreatic cancer by mass spectrometry: a multi-institutional validation study

Abstract Background Invasive ductal carcinoma of the pancreas is one of the worst prognoses among common human malignancies. The poor outcome of pancreatic cancer patients seems to be attributable to difficulty in early detection. Methods Mass spectra of peptide were obtained from a total of 1314 plasma/serum samples, consisting 4 cohorts (namely Cohorts 1-4) obtained from 7 medical institutions participating in the “Third-Term Comprehensive Control Research for Cancer” conducted by the Ministry of Health, Labor and Welfare of Japan and one institute in Germany using orthogonal matrix assisted laser/desorption ionization time of flight (oMALDI-qQTOF-MS). The amino acid sequences and posttranslational modifications of marker proteins were identified by tandem mass spectrometry (MS/MS). Results Plasma apolipoprotein AII heterodimer (ApoAII-2; 17252 m/z), unglycosylated apolipoprotein CIII (ApoCIII-0; 8766 m/z), and their summated value differed between 112 pancreatic cancer patients and 103 healthy controls (Cohort 1) with high statistical significance [P = 1.36 × 10−21, P = 4.35 × 10−14, and P = 1.83 × 10−24 (Mann-Whitney U-test)] and area-under-curve (AUC) values of 0.877, 0.798, and 0.903, respectively. The significance was further verified in 2 retrospective cohorts [Cohort 2 (n = 103) and Cohort 3 (n = 163)] and a hospital-based prospective cohort [Cohort 4 (n = 833)]. Patients with invasive ductal adenocarcinoma of the pancreas, including those with stage-I disease, could be discriminated from healthy controls with the sensitivity of 97.6% (243/249) and specificity of 95.3% (122/128) by the combination of DUPAN-2, CA19-9, ApoAII-2, and ApoCIII-0. Interpretation Although the reduction of the 2 modified forms of plasma/serum apolipoproteins was not pancreatic cancer-specific, the accurate measurement of plasma/serum proteins had a high capability in detecting pancreatic cancer. This test is essentially safe and might be applicable to the primary mass screening of pancreatic cancer in asymptomatic populations. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4580.

Detection of early-stage pancreatic cancer by high-resolution mass spectrometry

Detection of early-stage pancreatic cancer by high-resolution mass spectrometry

Highly sensitive detection of early‐stage pancreatic cancer by multimodal near‐infrared molecular imaging in living mice

Pancreatic cancer is a serious disease with poor patient outcome, often as a consequence of late diagnosis in advanced stages. This is in large part due to the lack of diagnostic tools for early detection. To address this deficiency, we have investigated novel molecular near-infrared fluorescent (NIRF) in vivo imaging techniques in clinically relevant mouse models of pancreatic cancer. Genome wide gene expression profiling was used to identify cathepsin cystein proteases and matrix metalloproteinases (MMP) as targets for NIRF imaging. Appropriate protease activatable probes were evaluated for detection of early-stage pancreatic cancer in mice with orthotopically implanted pancreatic cancer cell lines. Mice with pancreatitis served as controls. Whole body in vivo NIRF imaging using activatable cathepsin sensitive probes specifically detected pancreatic tumors as small as 1-2 mm diameter. Imaging of MMP activity demonstrated high specificity for MMP positive tumors. Intravital flexible confocal fluorescence lasermicroscopy of protease activity enabled specific detection of pancreatic tumors at the cellular level. Importantly, topical application of NIRF-probes markedly reduced background without altering signal intensity. Taken together, macroscopic and confocal lasermicroscopic molecular in vivo imaging of protease activity is highly sensitive, specific and allows discrimination between normal pancreatic tissue, inflammation and pancreatic cancer. Translation of this approach to the clinic could significantly improve endoscopic and laparoscopic detection of early-stage pancreatic cancer.