Chromogranin A as a Marker for Diagnosis,Treatment, and Survival in Patients With Gastroenteropancreatic Neuroendocrine Neoplasm

Chromogranin A (CgA) not only plays an important role in pathologic diagnosis, but is also used as a circulating biomarker in patients with gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN). However, the relationship between immunohistochemistry (IHC) expression and serum levels of CgA has not been investigated. The value of CgA for evaluating treatment response and prognosis is still not well understood. We conducted this study to assess the significance of CgA in GEP-NEN in terms of diagnosis, curative effects evaluation and prognosis.One hundred forty-five patients comprising 88 patients with active disease and 57 disease-free patients were enrolled in this study from January 2011 to November 2013. The expression of CgA was assessed by IHC, and serial serum CgA levels were measured by enzyme linked immunosorbent assay.The overall expression rate of CgA was 69.0% (100/145). CgA expression was associated with tumor site and stage (P < 0.05), but not correlated with prognosis (P = 0.07). Serum CgA levels were significantly higher in GEP-NEN patients with active disease when compared with disease-free patients (P = 0.001) or healthy participants (P < 0.001). A CgA cutoff value of 95 ng/ml discriminated between healthy subjects or disease-free patients and patients with active disease (sensitivity 51.2% and specificity 87.5%, respectively). There was a correlation between the CgA IHC expression and high serum CgA levels (R = 0.320, P = 0.002). Serum CgA levels were much higher in patients who classified as neuroendocrine carcinoma, mixed adenoendocrine carcinoma (P = 0.035) and who were on stage IV (P = 0.041). Changes in CgA levels normalization or ≥30% decrease suggested that patients had tumor response. Furthermore, patients with serum CgA levels higher than 95 ng/ml had a significantly shorter survival compared with patients with levels lower than 95 ng/ml (P < 0.001).CgA is a reliable pathologic and circulating maker for diagnosis of GEP-NEN. We further confirmed that serial measurement of CgA may be useful for evaluating the efficacy of different kinds of therapies in patients during follow-up, and serum CgA level ≥95 ng/ml may serve as a predictor of overall survial.     

Circulating MicroRNA Panel as a Diagnostic Marker for Hepatocellular Carcinoma

Background:Current diagnostic markers for hepatocellular carcinoma are compromised and limited by their low sensitivity and specificity. In this study, circulating microRNAs were utilized as a diagnostic tool to segregate hepatocellular carcinoma patients from healthy subjects.Methods:We analyzed 2 public datasets for differences in plasma microRNA expression profiles of hepatocellular carcinoma patients and healthy controls to identify biomarkers related to hepatocellular carcinoma. Plasma samples from hepatocellular carcinoma patients and control subjects were then collected for next-generation microRNA sequencing analysis. The differential microRNAs obtained from the above 3 parts were intersected to obtain microRNAs that were significantly different between the 2 groups. We then analyzed 58 specimens, which come from hepatocellular carcinoma and the control group, for validation through a quantitative polymerase chain reaction. The diagnostic value of these differentially expressed miRNAs was assessed by receiver operating characteristic curve analysis.Results:The levels of miR-206 and miR-222 were significantly higher (P < .05) and the level of miR-126 was lower (P < .05) in patients with hepatocellular carcinoma than in healthy subjects. Receiver operating characteristic analysis established a powerful diagnostic accuracy when miR-206, miR-222, and miR-126 were combined (area under curve = 0.887), which was similar to that of the marker α-fetoprotein (area under curve = 0.889). When the microRNAs were combined with α-fetoprotein, the accuracy of hepatocellular carcinoma diagnostic potential was further improved (area under curve = 0.989).Conclusion:We identified 3 microRNAs significantly altered in the plasma of hepatocellular carcinoma patients and they can screen patients at risk of hepatocellular carcinoma.     

血浆嗜铬粒蛋白A对胃肠胰腺内分泌肿瘤的诊断价值

背景：嗜铬粒蛋白A（CgA）广泛存在于神经内分泌细胞中，其血浆水平升高提示存在神经内分泌来源的肿瘤。目的：确定血浆CgA诊断胃肠胰腺内分泌肿瘤的临界点，并探讨其诊断价值。方法：以酶联免疫吸附测定（EUSA）检测39例胃肠胰腺内分泌肿瘤和37例嗜铬细胞瘤患者的血浆C外水平，以30例非内分泌肿瘤消化疾病患者和30例正常人作为对照。绘制接受者操作特征曲线（ROC曲线），确定ROC曲线下面积（AUROC）和血浆C小的诊断临界点。结果：胃肠胰腺内分泌肿瘤、嗜铬细胞瘤和非内分泌肿瘤消化疾病组的血浆CgA水平均显著高于正常对照组（86、135和30U／L对20U／L，P〈0．001）。以正常对照组和胃肠胰腺内分泌肿瘤组数据绘制ROC曲线，AUROC为0．912．CgA的诊断临界点为30U／L，敏感性为80．0％，特异性为96．7％；以正常对照组和嗜铬细胞瘤组数据绘制ROC曲线，AUROC为0．914，CgA的诊断临界点为30．5U／L，敏感性为89．2％，特异性为96．7％。结论：血浆C外水平对神经内分泌肿瘤，尤其是胃肠胰腺内分泌肿瘤具有较高诊断价值，可作为可靠的肿瘤标志物应用于临床。     

血清粗纤维调节素检测对诊断肝纤维化的临床价值

我们在提纯人胎盘粗纤维调节素(Un)及制备其抗血清的基础上,建立了竞争抑制ELISA法,检测143例各种肝病、30例非肝疾病和60名正常人血清Un水平,并与肝活检病理及血清Ⅲ型前胶原肽(PⅢNP)、透明质酸(HA)和层粘连蛋白(Ln)水平进行比较。结果各肝病组血清Un含量均显著高于正常对照及非肝疾病组(P值均<0.01):慢性肝炎各期间均有显著性差异(P值均<0.01);血清Un水产与慢性肝炎肝组织学炎症活动度(r=0.52,P<0.05)及纤维化程度(r=0.78,P<0.001)均呈显著正相关,与血清PⅢNP、LN、HA水平均不相关。若以血清Un≥200μg/L为判断肝纤维化的界值,其敏感性为71.43％,特异性为78.9％,诊断准确率为73.03％。     

Chromogranin A as potential target for immunotherapy of malignant pheochromocytoma

Currently, no effective treatment for malignant pheochromocytoma exists. The aim of our study was to investigate the role of chromogranin A (CgA) as a specific target molecule for immunotherapy in a murine model for pheochromocytoma. Six amino acid-modified and non-modified CgA peptides were used for dendritic cell vaccination. Altogether, 50 mice received two different CgA vaccination protocols; another 20 animals served as controls. In vitro tetramer analyses revealed large increases of CgA-specific cytotoxic T cells (CTL) in CgA-treated mice. Tumors of exogenous applied pheochromocytoma cells showed an extensive infiltration by CD8+ T cells. In vitro, CTL of CgA-treated mice exhibited strong MHC I restricted lysis capacities towards pheochromocytoma cells. Importantly, these mice showed strongly diminished outgrowth of liver tumors of applied pheochromocytoma cells. Our data clearly demonstrate that CgA peptide-based immunotherapy induces a cytotoxic immune response in experimental pheochromocytoma, indicating potential for therapeutic applications in patients with malignant pheochromocytoma.     

Probiotics in Experimental Ulcerative Colitis: Mast Cell Density and Neuronal Hypertrophy

Background:Probiotics such as Lactobacillus and Bifidobacterium are among the supportive treatment methods to achieve effective results in ulcerative colitis. This study was established to investigate the effect of probiotics in experimental ulcerative colitis and to detect changes in mast cell and neuronal structures in this treatment method.Methods:A total of 48 adult male rats were used to study the effects of probiotics on ulcerative colitis. The animals were divided into 6 groups as control, experimental colitis, and four probiotic protective groups. Three different bacterial strains were administered to the protective groups individually and in combination by gavage. PGP 9.5 antibody and mast cell tryptase were used for the detection of neuronal structures and mast cells. The number of Schwann cells and ganglia, size measurements of ganglia, and density of mast cells were evaluated.Results:Compared to the control, an increase in the number of mast cells was detected in all groups. Especially the increase in the number of mast cells was found to be statistically significant in combined probiotic administration. In the detection of neuronal structures, a significant increase in the number of Schwann cells and ganglia was detected in groups where probiotics were administered combined and individually.Conclusion:These results suggest that probiotics may play a role in the supporting effect of increasing the number of mast cells and neuronal structures, protecting the intestinal wall. We think that more specific and detailed studies should be conducted to evaluate the protective/therapeutic effect of probiotics in future studies.     

Surface Bound Immunoglobulins as a Cell Marker in Human Lymphoproliferative Diseases

Membrane-bound Ig were studied byimmunofluorescence methods in 116patients with lymphoproliferative disorders. Surface Ig synthetized in vitrowere studied in many patients. In 70of 73 cases of chronic lymphocyticleukemia (49 without and 24 withmonoclonal serum Ig), monoclonal surface Ig with a distribution for heavyand light chains similar to that of normal lymphocytes were found. In threecases, these surface Ig accumulatedas crystals in the lymphocyte cytoplasm. Studies limited to staining offreshly drawn cells may lead to erroneous conclusions since serum antibodies may become bound to the cellsurface and simultaneous positivity forµ, , , and , due either to the attachment of immune complexes at thelymphocyte surface or to anti-IgGantibody activity of the monoclonalsurface IgM, was not rare. IgM withanti-IgG activity was found on thesurface of both lymphocytes andlymphoblasts in a patient with acutetransformation of chronic lymphocyticleukemia. In a single case, µ and chains were simultaneously found onsome lymphocytes besides two singleproducer clones. A biclonal proliferation characterized by distinct surfaceIg markers was demonstrated in threeother cases of lymphocytic leukemiaand in a patient with Waldenström’smacroglobulinemia. In this latter condition (31 cases), most marrow lymphoidcells including plasma cells and themajority of blood lymphocytes carriedIgM determinants which had the samelight chain type as the serum IgM andwhich shared its eventual antibody activity. Similar results were obtainedin a few patients with the hematological features of macroglobulinemia butwith serum monoclonal IgG or IgA orwith unreleased IgM. Studies on casesof and heavy chain diseases, coldagglutinin disease, leukemic reticuloendotheliosis, and Sezary’s reticulosisare also recorded. The value of surfaceIg as B cell markers in lymphoproliferative diseases is outlined.

Submitted on June 6, 1972 Accepted on July 17, 1972     

Chromogranin A as serum marker of pituitary adenomas

OBJECTIVE: The diagnostic impact of chromogranin A (CgA) measurement has been studied in various neuroendocrine tumours (NET) such as pheochromocytomas, gastrinomas and neuroblastomas. Clinically nonfunctioning pituitary adenomas (NFPA) are generally diagnosed on tumoural symptoms or hypopituitarism and, except for gonadotrophins and their free subunits which may be increased in the case of gonadotrophinomas, markers of endocrine secretory activity are lacking not only for diagnostic purpose but also in the postoperative follow-up of these patients. As the presence of CgA has been demonstrated by immunohistochemistry in pituitary adenomas, we performed this study to further assess the sensitivity of CgA measurement in sporadic pituitary adenomas using a new, specific, sandwich immunoassay. SUBJECTS: We first completed a basal normative data set obtained using this assay by studying four healthy men (49 +/- 13 years old), five healthy premenopausal women (35.8 +/- 7.5 years old) and five healthy postmenopausal women (49.1 +/- 4.6 years old) basally and after TRH administration. Twenty-seven patients [12 men (64.2 +/- 11.8 years), even premenopausal women (38.4 +/- 5.7 years) and eight postmenopausal women (67.7 +/- 10.3 years)] with NFPA, 15 acromegalic patients [nine men (45 +/- 13.3 years), six women (52 +/- 14.9 years)] and 19 patients with a prolactin-secreting adenoma [four men (41.2 +/- 18 years) and 15 women (31.2 +/- 7.5 years), with a macroadenoma (n = 11) or a microadenoma (n = 8)] had basal and TRH-stimulated measurement of CgA. A gonadotrophin-releasing hormone (GnRH)-stimulation test was also performed in two, four and four patients, respectively. All patients had sporadic pituitary adenomas. MEASUREMENTS: Serum CgA was measured using a solid-phase two-site immunoradiometric assay based on monoclonal antibodies that bind to two distinct contiguous epitopes within the 145-245 region of CgA. RESULTS: Mean basal CgA concentration in 14 normal subjects was 80.2 ng/ml (SD: 31.7; range 19-124). A cut-off value for normal range was thus set at 125 ng/ml. TRH injection did not change significantly the CgA levels, peak values remaining less than 124 ng/ml. Three out of 27 subjects with NFPA (11%) had elevated basal CgA levels (576, 143, 241 ng/ml, respectively). Serum levels of CgA were not influenced by TRH in any of the NFPA subjects (including those three with increased basal levels). One out of 15 acromegalic patients (6.6%) and one out of 19 hyperprolactinemic patients (5.2%) had elevated serum basal CgA which did not significantly increase after TRH administration. In the remaining patients TRH-tests did not modify CgA levels. GnRH administration did not modify CgA levels. CONCLUSIONS: CgA serum levels measurement, assessed with a novel assay, does not provide a helpful marker for the clinical management of functioning and NFPA.     

RDW as an Additional Marker in Inflammatory Bowel Disease/Undifferentiated Colitis

BACKGROUND: Anemia is a common complication in inflammatory bowel disease patients. We postulate that the distribution of lesions in Crohn's disease is more likely than ulcerative colitis to lead to malabsorption as an additional cause of anemia. RDW, a simple and inexpensive test could be an additional differentiating test. METHODS AND RESULTS: Retrospective review of 284 cases of which 156 cases were diagnosed with Crohn's disease and 128 cases were diagnosed with ulcerative colitis. There was a significant difference in the mean RDW between the Crohn's and the ulcerative colitis cases (14.9 vs. 14.3, P = .027). CONCLUSIONS: We conclude there is a statistical significance between the two groups though this may not represent a clinically significant difference. From our analysis we conclude that RDW is statistically significant and with the implementation of a more rigorous study design and analysis of further data RDW may prove to be a clinically effective marker in differentiating Crohn's disease from ulcerative colitis.     

GPI-Anchored Fibromodulin as a Novel Target in Chronic Lymphocytic Leukemia: Diagnostic and Therapeutic Implications

Background: We have previously reported the aberrant expression of Fibromodulin (FMOD) in patients with chronic lymphocytic leukemia (CLL). Although FMOD has been considered as a cytoplasmic or secretory protein, we discovered the cell surface expression of FMOD in leukemic B cells via anchoring with glycosylphosphatidylinositol (GPI). Objective: To evaluate FMOD as a new biomarker in CLL patients in comparison with healthy individuals. Methods: A monoclonal antibody was generated against human FMOD. The cell surface expression of FMOD in 52 CLL patients and 45 healthy individuals were compared by flow cytometry. A bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) was used to determine the cell surface localization of FMOD using ELISA and flow cytometry techniques. Annexin V-FITC and propidium iodide (PI) was used to detect apoptosis induction in CLL PBMCs following in vitro incubation with anti-FMOD mAb. Results: The results demonstrated the widespread cell surface expression of GPI-anchored FMOD in CLL patients (median: 79.9 %), although healthy individuals had low FMOD expression (median: 6.2 %) (p≤0.0001). The cut-off value of FMOD expression was estimated with high sensitivity and specificity at 17.9%. Furthermore, in vitro apoptosis induction of leukemic cells following incubation with anti-FMOD mAb showed a direct apoptosis of CLL cells (27.9%) with very low effect on healthy PBMCs (6%). Conclusion: The membrane-anchoring of FMOD by means of a GPI moiety in leukemic cells supports FMOD as a highly potential diagnostic and therapeutic target in CLL patients.     

Limited role of Chromogranin A as clinical biomarker for pancreatic neuroendocrine tumors

BackgroundSerum Chromogranin A (CgA) is widely used as a biomarker for pancreatic neuroendocrine tumors (PanNETs). The aim of this study was to investigate the value of CgA as a diagnostic and prognostic marker for well-differentiated PanNETs.MethodsPatients with well-differentiated PanNET and a baseline CgA measurement, between 2011 and 2016 were reviewed. The diagnostic value was determined by comparing CgA values from patients with PanNETs to those with other pancreatic neoplasms and healthy controls. The Kaplan–Meier method was used to investigate the CgA prognostic significance.ResultsNinety-nine patients met inclusion criteria. As a diagnostic marker, CgA had a sensitivity of 66%, specificity of 95%, and overall accuracy of 71%. The use of PPIs was associated with a higher CgA level (p = 0.015). When excluding patients on PPIs, CgA accuracy in distinguishing PanNETs from other pancreatic neoplasms was 66%, the sensitivity and specificity were 60% and 75% respectively. Elevated CgA (p = 0.004), Ki67% (p < 0.001), tumor grade (p < 0.001) and stage of disease (p = 0.036) were associated with disease-specific survival.ConclusionCgA has a limited role as a diagnostic biomarker for well-differentiated PanNETs. An elevated CgA level may have prognostic value but its role should be further investigated with respect to other known pathological factors.     

Core Microbiota in Central Lung Cancer With Streptococcal Enrichment as a Possible Diagnostic Marker

BackgroundDysbiosis in lung cancer has been underexplored. The aim of this study was to define the bacterial and fungal microbiota of the bronchi in central lung cancer and to compare it with that of the oral and intestinal compartments.MethodsTwenty-five patients with central lung cancer and sixteen controls without antimicrobial intake during the previous month were recruited. Bacterial and fungal distribution was determined by massive sequencing of bronchial biopsies and saliva and faecal samples. Complex computational analysis was performed to define the core lung microbiota.ResultsAffected and contralateral bronchi of patients have almost identical microbiota dominated by Streptococcus, whereas Pseudomonas was the dominant genera in controls. Oral and pulmonary ecosystems were significantly more similar in patients, probably due to microaspirations. Streptococcal abundance in the bronchi differentiated patients from controls according to a ROC curve analysis (90.9% sensitivity, 83.3% specificity, AUC = 0.897). The saliva of patients characteristically showed a greater abundance of Streptococcus, Rothia, Gemella and Lactobacillus. The mycobiome of controls (Candida) was significantly different from that of patients (Malassezia). Cancer patients’ bronchial mycobiome was similar to their saliva, but different from their contralateral bronchi.ConclusionsThe central lung cancer microbiome shows high levels of Streptococcus, and differs significantly in its composition from that of control subjects. Changes are not restricted to tumour tissue, and seem to be the consequence of microaspirations from the oral cavity. These findings could be useful in the screening and even diagnosis of this disease.     

Chromogranin A as a marker of neuroendocrine neoplasia: an Italian Multicenter Study

Elevated circulating chromogranin A (CgA) levels are found in neuroendocrine tumors (NETs), but the diagnostic usefulness of this marker is still debatable. To assess the role of CgA for the diagnosis of gastroenteropancreatic (GEP) NETs and the identification of metastatic patients, an Italian multicenter observational study has been performed. CgA was evaluated in 202 GEP NET patients by IRMA and ELISA. The cutoffs for diagnosis and presence of metastases were identified by receiver-operating characteristic (ROC) curve. We found good correlation between IRMA and ELISA. The ROC analysis identified a cutoff of 53 ng/ml for IRMA and 16 U/l for ELISA as discriminating between controls and patients with active disease (sensitivity 71.3 and 84%; specificity 71 and 85% respectively). Metastases were present in 123 patients, having significantly higher CgA levels than patients without metastases. ROC analysis identified a cutoff of 146 ng/ml for IRMA and 67.3 U/l for ELISA as discriminating between patients with and without metastases (sensitivity 57 and 63.3%; specificity 55.6 and 71.4% respectively). For pancreatic NETs positive and negative predictive values were 84 and 78% respectively (90% specificity and 68% sensitivity). We found lower CgA levels in patients with extensive metastatic spread than in those with liver metastases only. These data assess the role of CgA evaluation in GEP NETs, and demonstrate that higher CgA levels associate with metastatic disease, confirming that CgA levels can provide a helpful practical biochemical marker for the clinical management of NETs, but with low sensitivity and specificity.