Serum Of Pancreatic Cancer Patients Research Articles

IRTIDP: A simple integrated real-time isolation and detection platform for small extracellular vesicles Glypican-1 in pancreatic cancer patients

Glypican-1 (GPC-1) protein-positive small extracellular vesicles (GPC-1+-sEV) have been proposed as potential biomarkers for early diagnosis of pancreatic cancer. In this study, we present an integrated real-time isolation and detection platform (IRTIDP) to capture and analyze GPC-1+-sEV directly from sera of pancreatic cancer patients. First, CD63 antibody-modified metal-organic framework (MOF) materials were utilized to enrich sEVs with a capture efficiency of 93.93 %. Second, a SERS probe was constructed by Raman reporter 4-MBA and GPC-1 antibody modified SERS active silver nanoparticles (AgNPs), which formed a sandwich complex structure of "MOFs@GPC-1+-sEV@AgNPs-4-MBA" with MOFs-enriched sEVs. The IRTSDP can complete the capture and detection process within 35 min, with a detection limit for 1 GPC-1+-sEV/μL, and linear range between 105∼109 GPC-1+-sEV/mL. Furthermore, this approach has been applied to quantify serum sEV GPC-1 in clinical pancreatic cancer patients. Based on the SERS intensity analysis, pancreatic cancer patients can be distinguished from pancreatic cystadenoma patients and healthy individuals effectively using this innovative platform that provides highly specific and sensitive means for early diagnosis of pancreatic cancer as well as other tumor types.

Detection of Tumor-Associated Autoantibodies in the Sera of Pancreatic Cancer Patients Using Engineered MUC1 Glycopeptide Nanoparticle Probes.

Pancreatic cancer is one of the deadliest cancers worldwide, mainly due to late diagnosis. Therefore, there is an urgent need for novel diagnostic approaches to identify the disease as early as possible. We have developed a diagnostic assay for pancreatic cancer based on the detection of naturally occurring tumor associated autoantibodies against Mucin-1 (MUC1) using engineered glycopeptides on nanoparticle probes. We used a structure-guided approach to develop unnatural glycopeptides as model antigens for tumor-associated MUC1. We designed a collection of 13 glycopeptides to bind either SM3 or 5E5, two monoclonal antibodies with distinct epitopes known to recognize tumor associated MUC1. Glycopeptide binding to SM3 or 5E5 was confirmed by surface plasmon resonance and rationalized by molecular dynamics simulations. These model antigens were conjugated to gold nanoparticles and used in a dot-blot assay to detect autoantibodies in serum samples from pancreatic cancer patients and healthy volunteers. Nanoparticle probes with glycopeptides displaying the SM3 epitope did not have diagnostic potential. Instead, nanoparticle probes displaying glycopeptides with high affinity for 5E5 could discriminate between cancer patients and healthy controls. Remarkably, the best-discriminating probes show significantly better true and false positive rates than the current clinical biomarkers CA19-9 and carcinoembryonic antigen (CEA).

Detection of Tumor‐Associated Autoantibodies in the Sera of Pancreatic Cancer Patients Using Engineered MUC1 Glycopeptide Nanoparticle Probes

AbstractPancreatic cancer is one of the deadliest cancers worldwide, mainly due to late diagnosis. Therefore, there is an urgent need for novel diagnostic approaches to identify the disease as early as possible. We have developed a diagnostic assay for pancreatic cancer based on the detection of naturally occurring tumor associated autoantibodies against Mucin‐1 (MUC1) using engineered glycopeptides on nanoparticle probes. We used a structure‐guided approach to develop unnatural glycopeptides as model antigens for tumor‐associated MUC1. We designed a collection of 13 glycopeptides to bind either SM3 or 5E5, two monoclonal antibodies with distinct epitopes known to recognize tumor associated MUC1. Glycopeptide binding to SM3 or 5E5 was confirmed by surface plasmon resonance and rationalized by molecular dynamics simulations. These model antigens were conjugated to gold nanoparticles and used in a dot‐blot assay to detect autoantibodies in serum samples from pancreatic cancer patients and healthy volunteers. Nanoparticle probes with glycopeptides displaying the SM3 epitope did not have diagnostic potential. Instead, nanoparticle probes displaying glycopeptides with high affinity for 5E5 could discriminate between cancer patients and healthy controls. Remarkably, the best‐discriminating probes show significantly better true and false positive rates than the current clinical biomarkers CA19‐9 and carcinoembryonic antigen (CEA).

TiRNA-Val-CAC-2 interacts with FUBP1 to promote pancreatic cancer metastasis by activating c‑MYC transcription

Cumulative studies have established the significance of transfer RNA-derived small RNA (tsRNA) in tumorigenesis and progression. Nevertheless, its function and mechanism in pancreatic cancer metastasis remain largely unclear. Here, we screened and identified tiRNA-Val-CAC-2 as highly expressed in pancreatic cancer metastasis samples by tsRNA sequencing. We also observed elevated levels of tiRNA-Val-CAC-2 in the serum of pancreatic cancer patients who developed metastasis, and patients with high levels of tiRNA-Val-CAC-2 exhibited a worse prognosis. Additionally, knockdown of tiRNA-Val-CAC-2 inhibited the metastasis of pancreatic cancer in vivo and in vitro, while overexpression of tiRNA-Val-CAC-2 promoted the metastasis of pancreatic cancer. Mechanically, we discovered that tiRNA-Val-CAC-2 interacts with FUBP1, leading to enhanced stability of FUBP1 protein and increased FUBP1 enrichment in the c-MYC promoter region, thereby boosting the transcription of c-MYC. Of note, rescue experiments confirmed that tiRNA-Val-CAC-2 could influence pancreatic cancer metastasis via FUBP1-mediated c-MYC transcription. These findings highlight a potential novel mechanism underlying pancreatic cancer metastasis, and suggest that both tiRNA-Val-CAC-2 and FUBP1 could serve as promising prognostic biomarkers and potential therapeutic targets for pancreatic cancer.

Abstract 371: Integrin-mediated lipolysis by pancreatic cancer-derived extracellular vesicles in cancer-associated cachexia

Abstract Introduction: Cancer-associated cachexia (CAC) is a life-threating condition frequently observed in cancer patients, especially those with pancreatic cancer. CAC is a multifactorial disease, with the notable characteristic of systemic lipolysis. Although humoral factors have been implicated in its pathogenesis, critical factors remain to be determined. Since even localized pancreatic cancer frequently induce lipolysis systemically, we examined the molecular mechanisms in vitro and in vivo based on the hypothesis that extracellular vesicles (EVs) systemically released from pancreatic cancer cells induce lipolysis. Procedures: EVs derived from the supernatants of cultured pancreatic cancer cell lines and from the sera of pancreatic cancer patients were characterized. The effects of EVs on adipocytes were determined using mature human adipocytes induced from human adipose-derived mesenchymal stem cells. To confirm the in vitro findings, lipolysis induction was examined in mice by intravenous injection of EVs. EV characteristics inducing lipolysis were determined. Those characteristics were confirmed by knockout studies and examining specifically isolated pancreatic cancer-derived EVs from heterogenous EV population in patients’ sera. Results: Lipolysis was triggered by specific EVs from pancreatic cancer cell line via the cAMP inside and uptake of more EVs into adipocytes was crucial for higher levels of lipolysis. In vivo studies, database search, and knockout studies revealed that integrin β1 and integrin α6 on EVs were the determinants for the attachment and uptake of the EVs to adipocytes, resulting in inducing more lipolysis. There was no difference in the percentage of these integrins positive EVs between heterogenous EV population in cancer patients’ sera and EVs in cancer-free controls’ sera. However, pancreatic cancer cell-derived EVs specifically isolated by targeting CA19-9, an aberrant glycosylation, from heterogenous EV population in patients’ sera, confirmed that pancreatic cancer-derived EVs have more such integrins. Conclusions: Cancer cell-derived EVs in the sera of pancreatic cancer patients have more ITGB1 and ITGA6 levels, which may explain the systemic induction of lipolysis in cancer cachexia from the early stages and could be novel therapeutic targets against CAC. In addition, specific isolation of cancer-derived EVs in sera from heterogenous EV population is crucial for elucidating cancer-related pathogenesis. Citation Format: Chikako Shibata, Motoyuki Otsuka, Mitsuhiro Fujishiro. Integrin-mediated lipolysis by pancreatic cancer-derived extracellular vesicles in cancer-associated cachexia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 371.

Multicolor biosensor for hyaluronidase based on target-responsive hydrogel and etching of gold nanorods by H2O2

Hyaluronidase (HAase) is a potential tumor biomarker for diseases of the digestive tract and nervous system, the development of simple and sensitive techniques for HAase determination is urgent needed. Gold nanorods (Au NRs) can be etched by H2O2 with high efficiency and display color changing. In this work, a HAase-responsive hydrogel system had been designed and the amount of H2O2 spilled from the system had a close relationship with the amount of HAase, then the spilled H2O2 had been applied to etch Au NRs. The color change of the solution was used to realize semi-quantitative determination of HAase. Furthermore, the longitudinal peak shift of Au NRs had a linear correlation with the concentration of HAase in the range of 10–60 U/mL (within 40 min) and the limit of detection (LOD) was 3.8 U/mL (S/N = 3), which can be used to realize accurate quantitative analysis of HAase. The proposed method has been applied to monitor HAase in serum of pancreatic cancer patients with satisfied results.

Isolation of Trypsin from Serum of pancreatic Cancer Patients and determination some biochemical parameters

The research included isolation and purification of Trypsin from serum of patient with pancreatic cancer, and estimation its molecular weight by using the two techniques of gel filtration (using sephadex G-50) and electrophoresis (using SDS). Therefore; it was noted that the proteinous peak isolated from ammonium sulfate solution is saturated (65%) of the serum by using the two techniques gel filtration and electrophoresis had apparent molecular weight (27000) Dalton. The study included (40) patients with pancreas cancer their ages ranged between (30-70) years who attended Al-Jumhuri and Atomic Medicine Hospitals in Mosul; the laboratory examination included estimation levels of: Trypsin, Gamma Glutamyl transferase, Amylase, Paraoxonase ,Glutathione, Sialic acid and Ceruloplasmin. Results showed that level of Trypsin and Amylase in the serum decreased significantly in patients with pancreatic cancer in comparison with the control group, but levels of Gamma Glutamyl transferase, paraoxonase, Glutathione, Sialic acid and Ceruloplasmin increased significantly in patients with pancreatic cancer in comparison with the control group.

Effects of the Exposure of Human Non-Tumour Cells to Sera of Pancreatic Cancer Patients.

Pancreatic ductal adenocarcinoma (PDAC) has high metastatic potential. The “genometastasis” theory proposes that the blood of some cancer patients contains elements able to transform healthy cells by transferring oncogenes. Since findings on genometastasis in PDAC are still scarce, we sought supporting evidence by treating non-tumour HEK293T and hTERT-HPNE human cell lines with sera of PDAC patients. Here, we showed that HEK293T cells have undergone malignant transformation, increased the migration and invasion abilities, and acquired a partial chemoresistance, whereas hTERT-HPNE cells were almost refractory to transformation by patients’ sera. Next-generation sequencing showed that transformed HEK293T cells gained and lost several genomic regions, harbouring genes involved in many cancer-associated processes. Our results support the genometastasis theory, but further studies are needed for the identification of the circulating transforming elements. Such elements could also be useful biomarkers in liquid biopsy assays.

MicroRNA-125a-3p, -4530, and -92a as a Potential Circulating MicroRNA Panel for Noninvasive Pancreatic Cancer Diagnosis.

MicroRNA (miRNA) expression dysregulations in pancreatic ductal adenocarcinoma (PDAC) have been studied widely for their diagnostic and prognostic utility. By the use of bioinformatics-based methods, in our previous study, we identified some potential miRNA panels for diagnosis of pancreatic cancer patients from noncancerous controls (the screening stage). In this report, we used 142 plasma samples from people with and without pancreatic cancer (PC) to conduct RT-qPCR differential expression analysis to assess the strength of the first previously proposed diagnostic panel (consisting of miR-125a-3p, miR-4530, and miR-92a-2-5p). As the result, we identified significant upregulation for all the three considered miRNAs in the serum of PC patients. After that, a three-miRNA panel in serum was developed. The area under the receiver operating characteristic curves (AUC) for the panel were 0.850, 0.910, and 0.86, respectively, indicating that it had a higher diagnostic value than individual miRNAs. Therefore, we detected a promising three-miRNA panel in the plasma for noninvasive PC diagnosis (miR-125a-3p, miR-4530, and miR-92a-2-5p).

Application of Magnetic Nanoparticles for Rapid Detection and In Situ Diagnosis in Clinical Oncology.

Simple SummaryScreening, monitoring, and diagnostic methods in oncology are a critical part of treatment. The currently used clinical methods have limitations, most notably the time, cost, and special facilities required for radioisotope-based techniques. The use of magnetic nanoparticles is an alternative approach that offers faster analyses with safer materials over a wide range of oncological applications, such as the detection of cancer biomarkers and immunostaining. Furthermore, magnetic nanoparticles, such as superparamagnetic iron oxide nanoparticles, can detect sentinel lymph nodes for breast cancer in a clinical setting, as well as those for gallbladder cancer in animal models within a timeframe that would enable them to be used during surgery with a magnetic probe.Screening, monitoring, and diagnosis are critical in oncology treatment. However, there are limitations with the current clinical methods, notably the time, cost, and special facilities required for radioisotope-based methods. An alternative approach, which uses magnetic beads, offers faster analyses with safer materials over a wide range of oncological applications. Magnetic beads have been used to detect extracellular vesicles (EVs) in the serum of pancreatic cancer patients with statistically different EV levels in preoperative, postoperative, and negative control samples. By incorporating fluorescence, magnetic beads have been used to quantitatively measure prostate-specific antigen (PSA), a prostate cancer biomarker, which is sensitive enough even at levels found in healthy patients. Immunostaining has also been incorporated with magnetic beads and compared with conventional immunohistochemical methods to detect lesions; the results suggest that immunostained magnetic beads could be used for pathological diagnosis during surgery. Furthermore, magnetic nanoparticles, such as superparamagnetic iron oxide nanoparticles (SPIONs), can detect sentinel lymph nodes in breast cancer in a clinical setting, as well as those in gallbladder cancer in animal models, in a surgery-applicable timeframe. Ultimately, recent research into the applications of magnetic beads in oncology suggests that the screening, monitoring, and diagnosis of cancers could be improved and made more accessible through the adoption of this technology.

Serum Exosomal EphA2 is a Prognostic Biomarker in Patients with Pancreatic Cancer

BackgroundPancreatic cancer (PC) is one of the worst prognoses amongst all malignant diseases. It is therefore of great significance to identify biomarkers with predictive clinical value for the prognosis and recurrence of PC.MethodsIn our study, enzyme-linked immunosorbent assays (ELISA) were used to detect the expression of Exo-EphA2 in the serum of PC patients and controls. Kaplan–Meier curve and Cox regression analyses were used to evaluate the prognostic value of Exo-EphA2 expression in patients with primary and recurrent PC.ResultsThe level of serum Exo-EphA2 was significantly higher in the PC group when compared to that of the control group. High expression of Exo-EphA2 in PC was associated with shorter overall survival (OS) and proved to be a significant negative prognostic factor in the multivariate analysis (HR = 1.04, 95% CI: 1.00–1.09, P <0.001). Additionally, we found that the level of serum Exo-EphA2 in recurrent PC patients (first recurrence < 12 months) was positively correlated with the level of Exo-EphA2 at primary diagnosis. Multivariate analysis showed that a high expression of Exo-EphA2 in recurrent PC was associated with shorter recurrence-free survival (RFS) (HR = 1.41, 95% CI: 1.10–1.70, P < 0.001).ConclusionHigh expression of serum Exo-EphA2 represents a novel biomarker for a poor prognosis in PC patients.

Aberrant lactate dehydrogenase A signaling contributes metabolic signatures in pancreatic cancer

BackgroundPancreatic cancer (PC) has the lowest 5-year survival rate; therefore, new early screening methods and therapeutic targets are still urgently required. Emerging technologies such as metabolomic-based liquid biopsy may contribute to the field. We found aberrant lactate dehydrogenase A (LDHA) signaling to be an unfavorable biomarker for PC.MethodsA total of 9 genes of the glycolysis pathway were detected by enrichment analysis in the PC Gene Expression Omnibus (GEO) dataset. The relationship between LDHA/pyruvate kinase (PKM)/fructose biphosphate aldolase A (ALDOA)/glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and patient survival was analyzed by Kaplan-Meier plotting analysis of The Cancer Genome Atlas (TCGA). The detection of changing metabolites in the serum of PC patients was performed using a nuclear magnetic resonance (NMR) spectrometer.ResultsWe found LDHA was an independent predictor of overall survival (OS) in PC patients (P<0.001). Consistent with genetic aberrance of LDHA, we identified significant alterations in patients’ glycolysis-related metabolites, including upregulation of lactic acid and downregulation of pyruvic acid. A 0.956 area under the curve (AUC) was achieved using the combinative metabolites score of lactic acid, pyruvic acid, citric acid, and glucose to distinguish PC from healthy controls.ConclusionsAberrant LDHA signaling is an unfavorable biomarker for PC and consequential metabolic changes constitute potential diagnostic signatures of PCs.

Tellurium doped zinc imidazole framework (Te@ZIF-8) for quantitative determination of hydrogen peroxide from serum of pancreatic cancer patients

The tellurium doped zinc imidazole framework (Te@ZIF-8) is prepared by a two-step hydrothermal strategy for the electrochemical sensing of hydrogen peroxide. Material is characterized by transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). The electrochemical characterization of the MOF modified electrode is done by a three-electrode system. Electrochemical sensing of hydrogen peroxide is made by cyclic voltammetry, amperometry, and impedance measurements. Results demonstrate that Te@ZIF-8 shows a detection limit of 60 µM with linearity up to 0.98855. Material is stable to 1000 cycles with no significant change in electrochemical response. Amperometry depicts the recovery of hydrogen peroxide from human serum up to 101%. Impedance curve reveals the surface of Te@ZIF-8-GCE (glassy carbon electrode) as porous and rough and an interface is developed between analyte ions and the sensing material. Finally, the modified electrode is used for the quantitative determination of hydrogen peroxide from serum samples of pancreatic cancer patients, diagnosed with CA 19-9.

MiR-504 and miR-205 May be Used to Evaluate Prognosis and Chemotherapeutic Effect of Pancreatic Cancer Patients

To explore the value of miR-504 and miR-205 in evaluating the prognosis and chemo-therapeutic effect of pancreatic cancer (PC) patients. Ninety-six PC patients who came to our hospital from March 2015 to February 2017 were selected as the research group (RG), and 96 healthy subjects during the same period were considered as the control group (CG). Serum miR-504 and miR-205 of subjects in the two groups were detected and compared by Rt-PCR, and the miR-504 and miR-205 expression levels in serum of PC patients before and after chemotherapy were analyzed, so did the relationship between miR-504, miR-205 and prognosis of patients. The miR-504 and miR-205 expression levels in serum of PC patients were dramatically higher than those of healthy people (P[ 0.05), and the expression levels in serum of those in the chemotherapy-effective group were dramatically lower than those of those in the chemotherapy-ineffective group (P[ 0.05). Moreover, the 3-year mortality of patients with high expression of miR-504 and miR-205 was higher than that of those with low expression (P[ 0.05), and the miR-504 and miR-205 expression levels in dead patients were higher than those in living patients. miR-504 and miR-205 had high predictive value for poor prognosis of PC patients. Univariate analysis identified that their poor prognosis was related to pathological stage, lymph node metastasis, differentiation degree and expression of miR-504 and miR-205 (P[ 0.05). Cox multivariate analysis identified that pathological stage, lymph node metastasis, differentiation degree and expression of miR-504 and miR-205 were independent risk factors for poor prognosis (P[ 0.05).DLX6-AS1 is highly expressed in the serum of PC patients, and it can evaluate the chemotherapeutic effect and predict the prognosis. So, it may be used as a new target direction for evaluating the chemotherapeutic effect and predicting the prognosis of patients.

Long non-coding RNA HULC as a diagnostic and prognostic marker of pancreatic cancer.

BACKGROUNDLong non-coding RNA (lncRNA) is abnormally expressed in various malignant tumors. In recent years, it has been found that IncRNA HULC is increasingly expressed in pancreatic cancer tissues and is involved in the development and progression of pancreatic cancer. However, the clinical value of serum HULC in pancreatic cancer remains unclear, and there are few studies on how HULC regulates the biological function of pancreatic cancer cells.AIMTo determine the value of lncRNA HULC in the diagnosis and prognosis of pancreatic cancer, and its possible biological potential.METHODSSixty patients with pancreatic cancer and sixty patients with benign pancreatic diseases admitted to Xiangya Hospital, Central South University were assigned to the pancreatic cancer group and the benign disease group, respectively, and another 60 healthy subjects were enrolled as the normal group during the same period. HULC-siRNA and NC-siRNA were transfected into pancreatic cancer cells. Quantitative real-time polymerase chain reaction was performed to determine the expression of HULC in tissues, serum, and cells. Western Blot was carried out to determine the expression of β-catenin, c-myc, and cyclin D1 in cells, and the cell counting kit-8, flow cytometry, and Transwell assay were conducted to determine the proliferation, apoptosis and invasion of cells.RESULTSHighly expressed in the tissues and serum of pancreatic cancer patients, HULC showed good clinical value in distinguishing between patients with pancreatic cancer, patients with benign pancreatic diseases and healthy subjects. HULC was related to pathological parameters including tumor size, T staging, M staging and vascular invasion, and the area-under-the-curve for evaluating these four parameters was 0.844, 0.834, 0.928 and 0.818, respectively. Patients with low expression of HULC had a significantly higher 3-year overall survival (OS) and 5-year OS than those with high expression. T staging, M staging, vascular invasion, and HULC were independent prognostic factors affecting the 3-year OS of patients with pancreatic cancer. Inhibition of HULC expression prevented the proliferation and invasion of pancreatic cancer cells, promoted apoptosis, and inhibited the expression of Wnt/β-catenin signaling pathway-related proteins, β-catenin, c-myc, and cyclin D1. The Wnt/β-catenin signaling pathway agonist (LiCl) restored proliferation, apoptosis, and invasion of pancreatic cancer cells with inhibited expression of HULC.CONCLUSIONHULC is an effective marker for the diagnosis and prognosis of pancreatic cancer, which may affect the biological function of pancreatic cancer cells through the Wnt/β-catenin signaling pathway.

Cyclophilin B promotes cell proliferation, migration, invasion and angiogenesis via regulating the STAT3 pathway in non-small cell lung cancer

Lung cancer is the most common type of cancer and has become the leading cause of cancer-associated mortality worldwide. It has been reported that expression of Cyclophilin B was greatly elevated in the pancreatic cancer patient sera as compared with the healthy volunteer sera. This study aimed to investigate the role and regulatory mechanism of CypB in NSCLC progression. The expression levels of CypB was detected in NSCLC samples and cell lines by ELISA, western blot and immunohistochemistry assay. In addition, CCK8, colony formation, scratch and transwell assays were used to evaluate the proliferation, migration and invasion of A549 cells with CypB silencing. The expression of angiogenesis related proteins and pathway-related factors were detected by western blot. In NSCLC samples, CypB expression was upregulated. The expression of CypB was significantly reduced in the siRNA-cyclophilin B group. In addition, CypB silencing inhibited cell proliferation, migration and invasion. The expression of angiogenesis related proteins and pathway-related factors have also changed significantly. These findings suggested that CypB silencing may suppress the proliferation, invasion, migration and angiogenesis of A549 cells via inhibiting STAT3 pathway.

Study Levels of Some Biochemical Parameters in Serum of Pancreatic Cancer Patients

This study included an estimation for the levels of some biochemical parameters in the serum of those pancreatic cancer patients, by collecting (40) blood samples from patients their ages ranged (30-70) years compared with (55) blood samples of healthy persons as control group. The parameters included Super oxide dismutase, Amylase, Ribonuclease (acidic and alkaline), Arylesterase, Alkaline phosphatease, Carcinoembryonic antigen, carbohydrate antigen 19-9, Total lipid and Bilirubin Results of the Statistical analysis indicated a significant decrease in the activity of Amylase and Super oxide dismutase patients in comparison with the control group. There was a significant increase in the activity of Ribonuclease (acidic and alkaline), Arylesterase, Alkaline phosphatease, Carcinoembryonic antigen, carbohydrate antigen 19-9, Total lipid and Bilirubin in patients in comparison with the control group.

Clinical significance of monitoring KRAS in tissue and serum of pancreatic cancer patients.

286 Background: KRAS monitoring provides valuable information for early diagnosis and prediction of treatment outcome in colorectal cancer. KRAS mutation is observed in only half of colon cancer patients, whereas it is detected in 90% of pancreatic cancer patients. Therefore, investigating tumor DNA in serum by KRAS monitoring may be even more valuable in pancreatic cancer patients. In this study, we elucidated the clinical significance of KRAS monitoring in pancreatic cancer patients during treatment. Methods: KRAS in tumor tissues was analyzed for mutations by Scorpion ARMS or RASKET in 69 patients with pancreatic tumors. KRAS in serum was analyzed for mutations (G12D, G12V, G12C, G12A, G12S, G12R, G13D, Q61L, and Q61H) using droplet digital polymerase chain reaction in 58 patients who underwent the curative surgery (N = 39) or the chemotherapy (N = 19) and who had KRAS mutation in their tissues. Results: KRAS mutation in tumor tissues was detected in 62 of 69 patients (92.5%). These 62 patients showed significantly poorer prognosis (3years overall survival; 43.9%) than the seven patients without mutation (p = 0.03), who were all alive. Monitoring of KRAS in serum revealed KRAS mutation in 22 of 58 patients (37.9%). In patients who underwent the chemotherapy (N = 19), 2years OS of patients who detected KRAS mutation in serum (N = 9) was 0% and them which not detected it (N = 10) was 46.7% (p = 0.01). And in the curative resection group (N = 39), we detected KRAS mutations in serum among recurrent patients after surgery, but did not detect them among non-recurrent patients. Conclusions: KRAS mutation in serum could be a valuable predictive and prognostic biomarker in pancreatic cancer patients. Additionally, assessment of KRAS in tumor tissues may provide information about individual tumor biology. So, monitoring KRAS status of patients with pancreatic cancer may be useful of the treatment strategy.

Abstract A062: Using CA19-9 as a translational biomarker for sLea targeted agents MVT-5873 and MVT-1075 in cancer

Abstract Background: Sialyl Lewis A (sLea) modification occurs on transmembrane and secreted proteins in pancreatic ductal adenocarcinoma (PDAC) and other GI cancers, leading to a high level of expression on cancer cells compared to normal tissue. sLea, also known as CA19-9, can be detected in patient serum using a CA19-9 diagnostic assay, which detects shed or secreted sLea modified proteins. MVT-5873, a fully human antibody that specifically targets the sLea antigen, is in a phase 1 clinical trial for the treatment of CA19-9 positive disease as a monotherapy (MVT-5873), as an imaging agent (MVT-2163), and as a radioimmunotherapy (MVT-1075). Methods: To understand the relevance of CA19-9 as a stratification marker, both annotated primary as well as patient-derived xenograft (PDX) tissue microarrays (TMAs) were stained with MVT-5873. To evaluate sLea as a response biomarker, serum samples from patients in the MVT-5873 phase 1 clinical trial were analyzed for CA19-9 levels using the diagnostic assay, and compared to their MVT-5873 PK profile. Available samples were also characterized by ELISA assay to determine the representation of sLea modification on MUC family proteins. Results: For the human CRC TMA, &amp;gt;75% of tumor cores displayed moderate to high levels of positive staining. In the PDX array, 21% of non-small cell lung cancer (NSCLC), 50% of small cell lung cancer (SCLC), and 69% of CRC cores stained positive. Overall, staining was observed with similar frequency regardless of mutational status of the tumor, suggesting that CA19-9 expression is independent of KRAS, BRAF, PIK3CA, or MMR pathway mutations. Staining was also relatively unchanged in lung PDX cancer tumor cores that were rendered chemoresistant compared to their chemosensitive precursor. The serum CA19-9 analysis of PDAC patients treated with MVT-5873 demonstrated that detectable CA19-9 is inversely proportional to MVT-5873 drug levels when CA19-9 levels are &amp;gt;100 U/ml. CA19-9 is not detected after MVT-5873 treatment when CA19-9 levels are &amp;lt;100 U/ml, suggesting that MVT-5873 can fully occupy all serum CA19-9 at those levels. Taken together, these data indicate that the CA19-9 diagnostic assay may be used as a measure for target engagement, and imaging data from the MVT-2163 phase 1 trial confirmed that the drug can efficiently accumulate at tumor sites in a time-dependent manner in patients. When pancreatic cancer patient serum was analyzed for protein scaffold composition, Muc1 and Muc16 were found to be present. Conclusions: These data support the use of sLea targeted therapeutics such as MVT-5873 and MVT-1075 in CRC and lung cancer, including for patients with KRAS or other genetic lesions. Additionally, for patients treated with MVT-5873, data suggest that the serum CA19-9 assay is a potential measure of target occupancy and is worthy of further investigation. Citation Format: H. Toni Jun, Christian Lohrmann, Viola Allaj, Wolfgang Scholz, G. Jonah Rainey, Wolfgang A. Weber, Jason S. Lewis, John Poirier, Paul W. Maffuid. Using CA19-9 as a translational biomarker for sLea targeted agents MVT-5873 and MVT-1075 in cancer [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr A062.

MicroRNA-185 suppresses pancreatic cell proliferation by targeting transcriptional coactivator with PDZ-binding motif in pancreatic cancer.

The aim of the present study was to compare the expression of transcriptional coactivator with the PDZ-binding motif (TAZ) in pancreatic cancer (PC) patients, and to investigate the regulation mechanisms of TAZ in the proliferation of PC. PC tissues and matched peritumoral tissues, pancreatic juice and serum were collected from PC patients who underwent pancreatectomy between June 2012 and December 2015 at the Affiliated Hospital of Qingdao University (Qingdao, China). Pancreatic juice and serum were collected from patients with chronic pancreatitis as a control. The levels of taz mRNA expression in the samples were examined by reverse-transcription quantitative polymerase chain reaction, and the protein expression of TAZ was assessed by western blot analysis and ELISA. MicroRNAs (miRNAs) that regulate TAZ expression were also predicted by bioinformatics analysis and validated by dual luciferase reporter and rescue assays. In addition, the proliferation of PC cells was evaluated after transfection with TAZ small interfering RNA (siRNA) or its upstream miRNA agomir. Expression of TAZ was significantly increased in the PC tissues, pancreatic juice and serum of PC patients at the mRNA and protein levels compared with controls (P<0.05). Furthermore, TAZ was predicted and verified to be a target of miRNA (miR)-185, and miR-185 and TAZ were inversely expressed in samples from PC patients (P<0.05). In addition, TAZ siRNA or agomiR-185 transfection significantly inhibited human pancreatic adenocarcinoma cell proliferation (P<0.05). However, overexpression of TAZ in the agomiR-185 group rescued the inhibition (P<0.05). Finally, the expression of TAZ effector proteins, namely ankyrin repeat domain-containing protein and cysteine-rich 61, were upregulated in PC tissues (P<0.05), but repressed following transfection of PC cells with agomiR-185 (P<0.05). Thus, miR-185 may regulate the proliferation of PC by targeting TAZ, making it a promising diagnostic marker for PC.