Chromogranin Research Articles

A260 THE LACK OF CHROMOGRANIN-A MODIFIES THE GUT MICROBIOTA COMPOSITION AND REGULATES EXPERIMENTAL COLONIC INFLAMMATION

Abstract Background Ulcerative colitis (UC)is characterized by distinct changes in the gut microbiome and elevated chromogranin-A (CHGA) level, which seem to be a relevant pathogenetic mechanism.CHGA, a prohormone produced by enterochromaffin (EC) cells and cleaved into several bioactive peptides, regulates experimental colonic inflammation. In the rodent, intra-rectal infusion of catestatin, a Chga-derived peptide, alters the distal colonic microbial composition. However, the interplay between CHGA, as a pro-hormone, and the gut microbiome remains elusive. Aims in homoeostatic and pathophysiologic conditions, we investigated the functional consequences of the lack of Chgaon the distal colonic microbiota. Methods Acute colitis (5 % dextran sulfate sodium [DSS], 5 days) was induced in Chga-C57BL/6-deficient (Chga-/-) and wild-type (Chga+/+)mice. Feces and mucosa-associated microbiota (MAM) samples were collected and the V4 region of 16s rRNA was subjected to Miseq Illumina sequencing. Alpha diversity was calculated using Shannon’s diversity index. OTU abundances were summarized using the Bray-Curtis index and non-metric multidimensional scaling (NMDS) analysis to visualize microbiome similarities and a permutational analysis of variance (PERMANOVA) to test the significance of groups were performed respectively. Results In non-colitic homoeostatic condition, the absence of Chga (Chga-/) significantly increased the bacterial richness and modified the bacterial community composition at the genera level between the groups, represented by increased abundance of Lactobacillus species and reduced abundance of Helicobacter& Oscillospira species compared to Chga+/+mice in fecal and colonic MAM. Moreover, the absence of Chga (Chga-/-) resulted in a significant change in the alpha-diversity of fecal and colonic MAM compared to Chga+/+mice. DSS induced-colitis resulted in a significant microbial dysbiosis in Chga+/+mice, however, deletion of Chgaprotected against DSS-induced colitis and reduced the microbial dysbiosis, reduced the family of Rikenellaceaeand maintained the abundance of Bacteroides species, compared to wild-type (Chga+/+). Conclusions The lack of CHGA regulates the biodiversity and the composition of the colonic gut microbiota suggesting a cross-talk between the EC cell and the microbiome. Therefore, targeting CHGA could provide a novel therapeutic strategy by regulating the gut microbiome in physiological and pathophysiological conditions. Funding Agencies CIHR

Chromogranin A provides additional prognostic information in children with severe hand, foot, and mouth disease: A prospective observational study

ObjectiveSevere hand, foot, and mouth disease (HFMD) is associated with high mortality in children, and persistent sympathetic activation is a common presentation. The aim of this study was to prospectively investigate serum chromogranin A (CHGA) levels and their prognostic role in this condition. MethodsSerum CHGA, creatine kinase myocardial band (CK-MB), serum D-dimer, norepinephrine, blood glucose, lactate, and C-reactive protein levels, white blood cell (WBC) counts, usage of vasopressors, pediatric risk of mortality Ⅲ (PRISM-Ⅲ) scores, and viral etiology were measured upon pediatric intensive care unit (PICU) admission. The correlation between clinical outcomes and the indicators listed above were analyzed, and the ability of CHGA as a biomarker to predict mortality was evaluated. ResultsSerum CHGA levels were higher in the non-survivors group than in the survivors group (median (interquartile range): 434.8 (374.3–502.4) vs 183.3 (131.9–246.9) μg/l; p < 0.001) and were correlated with norepinephrine (r = 0.37. p < 0.001), blood glucose (r = 0.32, p = 0.001), lactate (r = 0.25, p = 0.009), WBC (r = 0.20, p = 0.039), and PRISM-Ⅲ scores (r = 0.748, p < 0.0001). Patients suffering neurogenic pulmonary edema, those infected with enterovirus A71, and those requiring more vasopressors had higher serum CHGA levels (median (interquartile range): 385 (239.9–488.8) vs 161 (115.6–222.9), 340.6 (190.6–436.0) vs 150.5 (112.1–210.0), 395.6 (209.1–487.0) vs 167.7 (110.5–240.5) μg/l, respectively; p < 0.0001). The CHGA level upon PICU admission in severe HFMD could be an independent risk factor for mortality (adjusted odds ratio 2.459, 95% confidence interval 1.054–5.906, p = 0.038) with high specificity (87.5%) and sensitivity (82.6%) (cut-off value at 339.6 μg/l). ConclusionsThe CHGA level in severe HFMD was found to be associated with cardiopulmonary failure. If measured upon PICU admission, CHGA may provide additional prognostic information in this disease.

Endocrinological effect of lavender aromatherapy on stressful visual stimuli

IntroductionAromatherapy is prominent in complementary and alternative medicine. Little endocrinological evidence, however, of the effects of aromatherapy has yet been presented. We used salivary stress marker chromogranin A (CgA) to examine the effects of lavender aroma on women watching a stressful video. MethodsHealthy female university students (n = 23) aged 20–22 years old were randomly assigned to two groups: an aroma group exposed to lavender and an unexposed control group. Both groups watched a stressful video for 10 min. During the protocol, the aroma group was exposed to lavender aroma. Samples of salivary chromogranin A (CgA) were collected immediately before and after watching the video, and at 5 and 10 min after that. ResultsIn the aroma group, the levels of CgA statistically significantly decreased throughout the experimental period. In the control group, there was no such change. ConclusionThe findings suggest that lavender aroma may reduce the stress effects of watching a stressful video.

Chromogranin A Analysis in the Differential Diagnosis Across Lewy Body Disorders.

Chromogranin A (CgA) is a general marker of gut endocrine cells, which are part of the "gut-brain axis" in Parkinson's disease (PD). We analyzed CgA as a marker of synaptic dysfunction to assess its role in the differential diagnosis across different Lewy body disorders. We analyzed the CgA levels in the cerebrospinal fluid (CSF) and serum from 54 patients covering the spectrum of Lewy body disorders [13 Parkinson's disease (PD), 17 Parkinson's disease dementia (PDD), 24 dementia with Lewy bodies (DLB)] and 14 controls using an ELISA. A positive correlation was noted between CSF and serum CgA levels (ρ= 0.47, 95% CI: 0.24 to 0.65, p < 0.0001). The highest values of CgA in CSF and in serum were measured in DLB and there was a significant difference between DLB and PDD (p = 0.03 and p = 0.004). The serum levels of CgA in controls achieved lower values compared to DLB (p = 0.006). There was a gradual increase in serum levels from PD to PDD and DLB. An inverse correlation was seen between the CSF level of CgA and Aβ42 (ρ = -0.296, 95% CI: -0.51 to -0.04, p = 0.02). The incorporation of CgA analysis as an additional biomarker may be useful in the diagnostic work-up of Lewy body dementia. CgA analysis may be relevant in distinguishing DLB from PDD patients and presumably early stages of PD. Our data on altered serum levels in DLB pave the way to the development of blood-based parameters for the differential diagnosis, which however needs to be confirmed in a prospective study.

Syntaxin 1: A Novel Robust Immunophenotypic Marker of Neuroendocrine Tumors.

Considering the specific clinical management of neuroendocrine (NE) neoplasms (NENs), immunohistochemistry (IHC) is required to confirm their diagnosis. Nowadays, synaptophysin (SYP), chromogranin A (CHGA), and CD56 are the most frequently used NE immunohistochemical markers; however, their sensitivity and specificity are less than optimal. Syntaxin 1 (STX1) is a member of a membrane-integrated protein family involved in neuromediator release, and its expression has been reported to be restricted to neuronal and NE tissues. In this study, we evaluated STX1 as an immunohistochemical marker of NE differentiation. STX1, SYP, CHGA, and CD56 expression was analyzed in a diverse series of NE tumors (NETs), NE carcinomas (NECs), and non-NE tumors. All but one (64/65; 98%) NETs and all (54/54; 100%) NECs revealed STX1 positivity in at least 50% of the tumor cells. STX1 showed the highest sensitivity both in NETs (99%) and NECs (100%) compared to CHGA (98% and 91%), SYP (96% and 89%), and CD56 (70% and 93%), respectively. A wide variety of non-NE tumors were tested and found to be uniformly negative, yielding a perfect specificity. We established that STX1 is a robust NE marker with an outstanding sensitivity and specificity. Its expression is independent of the site and grade of the NENs.

Molecular identification of bronchopulmonary neuroendocrine tumours and neuroendocrine genotype in lung neoplasia using the NETest liquid biopsy.

OBJECTIVESDiagnosing lung neuroendocrine neoplasia (NEN) requires a biopsy or an operation. We evaluated a ‘liquid biopsy’ (NETest) as an in vitro diagnostic tool for identifying NEN and compared it to chromogranin A (CgA).METHODSWe identified 4 study cohorts: patients with bronchopulmonary carcinoids (n = 99, including 62 typical and 37 atypical carcinoids), lung cancers [n = 101, including 41 adenocarcinomas, 37 squamous carcinomas (SQC), 16 small-cell lung cancers and 7 large-cell neuroendocrine carcinomas]; benign disease (50 idiopathic pulmonary fibrosis) and healthy controls (n = 102). Transcript levels measured quantitatively (activity scores: 0–100) were compared to CgA (enzyme-linked immunosorbent assay; normal < 109 ng/ml) levels.RESULTSThe results of the NETest were positive (>20) in 94% of patients with bronchopulmonary carcinoid compared to 8% of the controls (Fisher’s exact test; P < 0.001) and were significantly more accurate as a diagnostic test (McNemar’s test; P < 0.001, χ2 = 72) than was CgA (positive: 19% bronchopulmonary carcinoid, 15% controls). Small-cell lung cancers (87%), large-cell neuroendocrine carcinomas (86%), adenocarcinoma (42%) and SQC (35%) were also NETest-positive. Increasing the NETest cut-off score to >40 was useful for detecting all NENs and differentiating these tumours from either controls/benign lung diseases (specificity 97%) or adenocarcinoma/SQC (specificity 94%). CgA was positive in 15–44% irrespective of pathology and had no diagnostic value.CONCLUSIONSA gene-based liquid biopsy is an effective and accurate method for diagnosing lung tumours with neuroendocrine gene expression. CgA has no value. An NETest score >40 provides an accurate (94–97%) rule-in for the diagnosis of NEN and a rule-out for benign and other neoplastic diseases. Because neuroendocrine gene expression is associated with a poor prognosis, NETest levels may have utility both in the diagnosis of and the treatment stratification for lung neoplasia.

Non-toxic fragment of botulinum neurotoxin type A and monomethyl auristatin E conjugate for targeted therapy for neuroendocrine tumors.

Surgical resection is the only cure for neuroendocrine tumors (NETs). However, widespread metastases have already occured by the time of initial diagnosis in many cases making complete surgical removal impossible. We developed a recombinant heavy-chain receptor binding domain (rHCR) of botulinum neurotoxin type A that can specifically target synaptic vesicle 2 (SV2), a surface receptor abundantly expressed in multiple neuroendocrine tumors. Expression of neuroendocrine differentiation markers chromogranin A (CgA) and achaete-scute complex 1 (ASCL1) were signficantly reduced when treated with rHCR. rHCR conjugated to the antimitotic agent monomethyl auristatin E (MMAE) significantly suppressed proliferation of pancreatic carcinoid (BON) and medullary thyroid cancer cells (MZ) at concentrations of 500 and 300 nM respectively, while no growth suppression was observed in pulmonary fibroblasts and cortical neuron control cell lines. In vivo, rHCR-MMAE significantly reduced tumor volume in mouse xenografts with no observed adverse effects. These data suggest recombinant HCR (rHCR) of BoNT/A preferentially targets neuroendocrine cancer without the neurotoxicity of the full BoNT/A and that SV2 is a specific and promising target for delivering drugs to neuroendocrine tumors.

Significance of chromogranin A and synaptophysin in pancreatic neuroendocrine tumors.

The two most commonly used immunohistochemical markers for neuroendocrine cells and their tumors are chromogranin A (CgA) and synaptophysin (SPY). CgA is a marker for neuroendocrine secretory granules of four pancreatic hormones and gastrin while SPY is a marker for synaptic vesicles in neuroendocrine cells, which release classic neurotransmitters such as acetylcholine and others. CgA is involved in synthesis and secretion of peptide hormones through exocytosis while the function of SPY is elusive. Thirty-five pancreatic neuroendocrine tumors (Pan-NETs) were studied, consisting of 14 insulinomas, 8 gastrinomas, 2 glucagonomas, 6 pancreatic polypeptidomas and 5 non-functioning tumors, and were immunostained for four pancreatic hormones, gastrin, CgA, and SPY. Majority of Pan-NETs were less immunostained for the endocrine hormones and CgA than the normal pancreatic endocrine cells. CgA immunostaining mostly correlates with each hormone staining in non-β-cell tumors, while SPY immunostaining recognizes endocrine cells diffusely in the cytoplasm. CgA immunostaining is less in insulinomas than in non-β-cell tumors, and CgA immunostaining may distinguish CgA-weaker insulinomas from CgA-stronger non-β-cell tumors. CgA immunostaining may be used as an independent marker for biological aggressiveness in non-β-cell Pan-NETs. The serum CgA levels are higher in subjects harboring non-β-cell tumors than those harboring insulinomas, and the serum CgA elevates in parallel to the increasing metastatic tumor mass. Thus, CgA positive immunostaining in Pan-NETs correlates with the elevated serum levels of CgA for diagnosing CgA-positive non-β-cell Pan-NETs and the increasing serum CgA levels indicate increasing metastatic tumor mass.

Serum chromograninA level continuously rises with the progression of type1 diabetes, and indicates the presence of both enterochromaffin-like cell hyperplasia and autoimmune gastritis.

Aims/IntroductionThe relationship of chromogranin A (CgA) levels above the normal range with various outcomes, such as glycated hemoglobin levels, enterochromaffin‐like cell hyperplasia and autoimmune gastritis, was investigated in type 1 diabetes patients with special regard to the progression of comorbidities.Materials and MethodsA cohort study on 153 type 1 diabetes patients was carried out with a prospective branch on clinical and laboratory data, and a retrospective branch on histological data obtained by gastroscopy.ResultsPatients with CgA levels above the upper limit of the normal range (n = 28) had significantly higher glycated hemoglobin levels (P = 0.0160) than those with CgA in the normal range (n = 125). The correlation between CgA and glycated hemoglobin was significant (P < 0.0001), but weak (R = +0.32). A slight, but steady elevation (P = 0.0410) in CgA level was observed to co‐vary with the duration of type 1 diabetes. Enterochromaffin‐like cell hyperplasia and autoimmune gastritis was significantly more frequent (P = 0.0087 for both) in the high CgA group. Detailed analyses on gastric tissue samples confirmed a progression of enterochromaffin‐like cell hyperplasia (P = 0.0192) accompanied by CgA elevation (P = 0.0316).ConclusionsThe early detection and follow up of the later progression of enterochromaffin‐like cell hyperplasia and autoimmune gastritis into gastric neuroendocrine tumors, which have ~100‐fold greater incidence in type 1 diabetes patients, can be achieved by assessment of CgA levels. Therefore, the use of CgA could be considered as a novel auxiliary biomarker in the care of these type 1 diabetes complications.

Blood neuroendocrine gene transcripts (NETest) as a diagnostic of pancreatic neuroendocrine tumors and to identify the efficacy of surgical resection.

631 Background: Surgery is the only treatment for cure of pancreatic neuroendocrine tumours (PNET). There are no effective biomarkers to assess completeness of resection and predict recurrence. The aims of this study were 1) evaluate if blood measurement of neuroendocrine gene transcripts (NETest) were diagnostic of PNET and 2) assess if blood levels correlated with the resection type (R0/ R1/R2). We compared transcript analysis with Chromogranin A (CgA), the current standard of care. Methods: A prospective, longitudinal, single center study (ClinicalTrials.gov ID: NCT03012789) of 30 PNETs. Blood for NETest and CgA were collected preop and on postop day 30 (POD30). Transcript measurement: real-time quantitative reverse transcription PCR and multianalyte algorithmic analysis (NETest) (normal 0-20, &gt; 20 stable and &gt; 40 progressive disease; CgA: (ELISA) (normal &lt; 109mg/ml). Analysis: Student’s t test, Mann-Whitney U test and Fisher’s exact test. Data: mean±SEM. Results: Surgical resections ( n= 30) were: 26 R0, 2 R1, 2 R2. In vitro diagnostic: Preop NETest positive in all 30 (NETest: 38±4) (1005) whereas CgA levels positive (220±66ng/ml) in 9 (30%, p&lt; 0.0001). Surgery R0: ( n= 26): NETest significantly decreased from pre-op of 45±3 to 21±4 ( p&lt; 0.001) at POD30. Post operatively 13 (50%) exhibited an elevated score. One patient (Ki67 18%; extensive nodal involvement) had a score of 80, consistent with residual progressive disease. The remaining 12 had NETest levels of 27 at POD 30. CgA was elevated in 8 (31%) with no decrease at POD30 (154±44ng/ml vs. 109±22ng/ml). R1/R2 ( n= 4): NETest scores unchanged (42±17 vs 46±16) at POD30. CgA levels unchanged (378±332 vs. 753±634ng/ml). Conclusions: The NETest is an accurate in vitro diagnostic biomarker for PNET (100%). After R0 resection NETest decrease indicates the blood test is concordant with tumor removal. Elevated post-resection scores are consistent with residual disease and with reported recurrence rates after pancreatic surgery. CgA exhibited no clinical utility. At 1 month post-surgery, the NETest provides an early indication of high risk recurrence patients who can be stratified for further therapy. Clinical trial information: NCT03012789.

A Systematic Review of Chromogranin a (CGA) and its Biomedical Applications, Unveiling its Structure-Related Functions

A Systematic Review of Chromogranin a (CGA) and its Biomedical Applications, Unveiling its Structure-Related Functions

Dedifferentiation of neuroendocrine carcinoma of the uterine cervix in hypoxia

Neuroendocrine carcinoma of small cell type (SCNEC) is a rare pathological subtype in cervical cancer, which has a worse prognosis than other histological cell types. Due to its low incidence and the lack of experimental platforms, the molecular characteristics of SCNEC in the cervix remain largely unknown. Using the cancer tissue-originated spheroid (CTOS) method—an ex vivo 3D culture system that preserves the differentiation status of the original tumors—we established a panel of CTOS lines of SCNEC. We demonstrated that xenograft tumors and CTOSs, respectively, exhibited substantial intra-tumor and intra-CTOS variation in the expression levels of chromogranin A (CHGA), a neuroendocrine tumor marker. Since hypoxia affects differentiation in various tumors and in stem cells, we also investigated how hypoxia affected neuroendocrine differentiation of SCNEC of the uterine cervix. In the CTOS line cerv21, hypoxia suppressed expression of the neuroendocrine markers CHGA and synaptophysin (SYP). Flow cytometry analysis using CD99 (a membrane protein marker of SCNEC) revealed decreased CD99 expression in a subset of cells under hypoxic conditions. These expression changes were attenuated by HIF-1α knockdown, and by a Notch inhibitor, suggesting that these molecules played a role in the regulation of neuroendocrine differentiation. The examined SCNEC markers were suppressed under hypoxia in multiple CTOS lines. Overall, our present results indicated that neuroendocrine differentiation in SCNEC of the uterus is a variable phenotype, and that hypoxia may be one of the factors regulating the differentiation status.

The associations between anxiety/depression and plasma chromogranin A among healthy workers: Results from EHOP study.

ObjectivesChromogranin A (CgA) is regarded as an indicator of sympathetic tone and adrenomedullary system activity. Catestatin is one of CgA‐derived fragments through proteolytic processing. Many studies have confirmed the correlation between anxiety/depression and the salivary CgA level. The study was to investigate the associations between anxiety/depression and plasma CgA/catestatin levels in healthy workers without cardiovascular disease.MethodsThe study included 263 healthy workers (175 men and 88 women). The symptoms of anxiety and depression were measured with the Hospital Anxiety and Depression Scale (HADS). Plasma CgA and catestatin levels were measured by ELISA kits.ResultsIn bivariate correlation analysis, anxiety and depression were positively associated with plasma CgA level, respectively (r = 0.298, P < .001; r = 0.304, P < .001), but not significantly associated with plasma catestatin level.The anxiety group had significantly higher plasma CgA level than that in the no‐anxiety group (median 158.60 vs 70.90, P < .001). The similar results were found for depression scales. The depression group had significantly higher plasma CgA level (median 145.60 vs 82.40, P < .001).In the multiple linear regression model, after adjusting for age, gender, and BMI, anxiety was positively correlated with plasma CgA level (β = 0.359, P < .001), while anxiety was negatively correlated with plasma catestatin level (β = −0.128, P = .044), depression was also positively correlated with plasma CgA level (β = 0.343, P < .001).ConclusionsPlasma CgA was associated with anxiety and depression in healthy workers. It can be considered as the blood indicator for the evaluation of anxiety and depression.

P53, Somatostatin receptor 2a and Chromogranin A immunostaining as prognostic markers in high grade gastroenteropancreatic neuroendocrine neoplasms

BackgroundHigh grade gastroenteropancreatic (GEP) neuroendocrine neoplasms (NEN) with a Ki67 proliferation index > 20%, include well-differentiated tumours grade 3 (NET G3) and poorly differentiated (PD) neuroendocrine carcinomas (NEC). Abnormal p53-expression is a feature of PD tumours, while expression of chromogranin A (CgA) and somatostatin-receptor 2a (SSTR-2a) may be a feature of well-differentiated tumours. The aim of this study was to elucidate the expression and prognostic value of these three markers in 163 GEP-NEN patients with a Ki67-index > 20%.MethodClinical data, histopathology and overall survival were analysed according to Kaplan-Meier’s method and Cox regression. The expression of SSTR-2a, CgA and synaptophysin was analysed in tumour specimens by immunohistochemistry, and semi-quantitatively scored as negative (< 5%), heterogeneously positive (5–30%) or strongly positive (> 30%). P53 was defined as normal when scored as heterogeneously positive (1–30%), and abnormal when negative (0%) or strongly positive (> 30%).ResultsIn multivariate analysis, better survival was observed among patients with heterogeneously positive p53 compared to strongly positive (p < 0.001). When dichotomised, tumours with a heterogeneously positive p53 vs. negative and strongly positive p53 also showed a significantly better survival (p = 0.002).Survival was significantly worse for negative CgA compared to heterogeneously positive CgA (p = 0.02). Strongly positive SSTR-2a expression was found in 26% of the 163 included patients. Well-differentiated morphology correlated with strong expression of SSTR-2a and CgA, and heterogeneously positive p53-staining, and was more frequent in pancreatic primaries. In pancreatic primaries, strongly positive SSTR-2a was associated with longer survival (univariate analysis, p = 0.02). A significantly lower Ki67 proliferation index was found in patients with a heterogeneously positive p53, a positive SSTR-2a and CgA expression.ConclusionOur results suggest that abnormal p53-expression is an independent negative prognostic marker in GEP-NEN with a Ki67-index > 20%. Patients with heterogeneously positive p53 had the best prognosis. SSTR-2a was a positive prognostic marker in pancreatic NEN. Negative CgA was associated with a significantly worse OS compared to heterogeneously positive CgA-expression in a multivariate sub-analysis. Lower Ki67 index correlated significantly with heterogeneously positive p53, positive SSTR-2a and CgA expression.

Chromogranin A and serotonin for evaluation of treatment efficacy of neuroendocrine tumors

Background: The utility of biochemical markers in the monitoring of treatment efficacy in patients with neuroendocrine tumors (NETs) goes beyond any doubt. However, there are still no clear criteria for the assessment of clinically significant abnormalities of the main NET biomarkers chromogranin A (CgA) and serotonin. Aim: To evaluate the value of serial measurement of serum CgA and serotonin in the monitoring of the treatment effect in NET patients. Materials and methods: Serum CgA and serotonin levels were measured in 107 patients with NETs at baseline and at 3–4 weeks after the end of treatment (chemotherapy, biotherapy), as well as in 60 healthy controls. We used enzyme immunoassay based on Chromogranin A NEOLISA (Euro Diagnostica) and Serotonin ELISA (IBL International GmbH) test systems. Results: There was an association between CgA levels and the efficacy of chemotherapy in NET patients. With progression of the disease, median CgA increased significantly from 412 to 2679 ng/mL (p = 0.012), whereas in the patients with partial response it decreased from 811 to 254 ng/mL (p = 0.023). The ROC analysis showed the 33% cut-off for significant CgA changes for progression, with sensitivity of 80.0% and specificity of 95.6%. A decrease (of more than 33% compared to baseline levels) or absence of significant CgA changes was associated with stabilization of the disease or with partial response to treatment. Significantly decreased CgA levels were found in 75.0% cases of partial response and 43.48% of stabilized patients, whereas the absence of any significant changes in 25 and 66.7%, respectively. There was no association between serotonin levels and the disease behavior under treatment.Conclusion: CgA could be used as a sensitive marker of NET progression on chemotherapy.

CD57 as a routine neuroendocrine marker for liver metastasis.

The characterization of hepatic metastases as having neuroendocrine origins is essential and the main markers currently used are chromogranin A (CgA) and synaptophysin (Syn). However, these markers may exhibit certain limitations, and the use of CD56 and CD57 can also be considered, although, due to low specificity, their use is discouraged. This study sought to compare the immunohistochemical expression of these markers in hepatic metastases of neuroendocrine neoplasms (NEN). Eighteen samples, were used for immunohistochemical staining with CgA, Syn, CD56, and CD57 antibodies. The immunostaining reactions were compared according to its intensity (I), the percentage of labeled cells (P), and a final score (I × P). Statistical agreement between the markers was also evaluated. CD57 was expressed in the highest number of cases and also showed the most intense expression. CgA showed the highest number of cases with more than 80% positively stained area (72.2%), followed by CD57 (61.1%). The highest average score (I × P) was obtained for CD57 (9.1 ± 4.1). The best indices of agreement were between CgA and CD57 with respect to positivity (P = 0.021) and score (P = 0.014). According to the primary site, stomach/duodenum, lungs, and undetermined subgroups showed the highest average scores for CD57, followed by CgA. For the small bowel subgroup, the highest average score was obtained for CgA, followed by CD57. Our results highlight the importance of CD57 in the evaluation of hepatic metastases of NEN and indicate that this marker should be included with CgA and Syn in routine diagnostic panels.

Clinical utility of serum chromogranin A as a marker of hepatocellular carcinoma in chronic hepatitis patients

<br><b>Background</b><br>Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world and the third most common cause of cancer deaths accounting for about 745 000 deaths per year globally. HCC is often diagnosed at an advanced stage where effective therapies are lacking.<br><b>Aim</b><br>Is to investigate the role of serum chromogranin A (CgA) as a marker for detection of HCC in patients with HCC complicating chronic hepatitis and correlate its serum levels with alpha fetoprotein (AFP).<br>This study was conducted on 90 HCC patients (group I), 30 patients with hepatic cirrhosis (group II), and 30 age-matched and sex-matched healthy-controls (group III). Group I was further subdivided into group Ia (30 patients) with HCC complicating chronic hepatitis B, and group Ib (60 patients) with HCC complicating chronic hepatitis C. All participants included in the study were subjected to full history taking and clinical examination, laboratory investigations, including complete blood count, liver-function tests, renal-function tests, and viral markers. Serum AFP and serum CgA levels were also determined.<br><b>Results</b><br>Our results revealed that there was a highly significant elevation (<i>P</i> < 0.01) in the median serum CgA in hepatitis-B virus -related HCC (200 ng/ml) and hepatitis-C virus-related HCC (190 ng/ml) when compared with the control group (30 ng/ml) and when compared with cirrhotic group (60 ng/ml) with no significant difference between the median serum CgA in hepatitis-B virus -related HCC and hepatitis-C virus-related HCC (<i>P</i> > 0.05). A highly significant positive correlation between chromogranin and AFP (<i>P</i> < 0.01) was found in HCC patients. On the other hand, no significant correlations were found between chromogranin and complete blood count, kidney functions, and liver-function tests in patient groups. Diagnostic reliability testing revealed that for CgA, the best cutoff for discriminating HCC patients was 100 ng/ml at which the sensitivity was 77.78%, the specificity 93.3% with positive predictive value 94.59% and negative predictive value 73.68%. Regarding the combination between CgA and AFP, it showed a sensitivity of 97.78% and a specificity of 86.67%.<br><b>Conclusion</b><br>Serum CgA is a promising sensitive and specific tumor marker for identification of HCC. Addition of serum CgA to the current standard tests will improve the sensitivity and accuracy of diagnosis of HCC patients and thus could allow them to benefit from earlier treatment.<br>

Association of the CHGA gene polymorphism in patients with hemorrhagic stroke and/or aneurysm

ABSTRACT Introduction: Cerebrovascular diseases have been associated with several genes. Chromogranin A (CHGA) has been used as maker in cardiovascular disease. Therefore, evaluating the polymorphism and verifying its association with this pathology is very important to better understand this disease. Objective: The aim of this study was to identify the association between coding region polymorphism in -264 position of the CHGA gene (Glu264Asp) and hemorrhagic stroke (HS)/aneurysm in the Federal District, Brazil. Methods: This is a population-based case-control, involving 45 cases with HS and/or aneurysm. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method is used for genotyping these samples. A significance level of 5% was adopted. Results: The absence of the CC genotype the Glu264Asp CHGA polymorphism in the study participants and the significant presence of the GC heterozygote genotype were observed in this study. However, the distribution of genotypes did not differ statistically in the groups. Conclusion: The Glu264Asp CHGA polymorphism does not seem to contribute to the genesis of the CHGA protein expression in this patients group, but to understand whether or not there is a possible association of the pathology in question and whether the mutation will contribute in the gene therapy and thus to improve patients’ quality of life.

Actinin-4 splice variant - a complementary diagnostic and prognostic marker of pancreatic neuroendocrine neoplasms.

Introduction: For pathological diagnosis of pancreatic neuroendocrine neoplasms (pNENs) the routinely used immunohistochemical markers are chromogranin A (CgA) and synaptophysin (Syn). Their ability as prognostic markers is not well established. A splice variant of actinin-4 (Actn-4sv) was recently found to be an excellent biomarker of neuroendocrine neoplasms of the lung. We aimed to investigate the expression of Actn-4sv in pNENs and evaluate its quality as a biomarker of pNENs.Methods: Paraffin-embedded and frozen tissues specimens from 122 pNENs were analyzed. Western blots were performed to prove and compare the relative amount of Actn-4sv expression in pNENs tissue homogenates. For comparison pancreatic ductal adenocarcinoma (PDAC) and normal pancreatic tissues were analyzed in parallel. Immunohistochemistry (IHC) of paraffin sections of pNENs for Actn-4sv were performed and compared to the classic neuroendocrine markers CgA and Syn. Correlations were calculated between the staining intensity and distribution of Actn-4sv and staging, grading and afflicted lymph nodes respectively.Results: Actn-4sv was expressed in 88.5% (108/122) of pNENs, but not in normal pancreatic tissues (0/14) or PDAC (0/14). Compared to CgA and Syn, Actn-4sv was not detectable in islet cells of the normal pancreas. Staining intensity of Actn-4sv on pNENs negatively correlated to the histological grading (Spearman r=-0.4990, p<0.0001) and staging (r = -0.2581, p = 0.0041) but no correlation to afflicted lymph nodes was found. A significantly better overall survival was observed for pNEN patients with higher expression of Actn-4sv (hazard ratio 2.7; log-rank test p= 0.0349).Conclusions: The expression of Actn-4sv may be an important prognostic factor for patients with pNENs. Its expression correlates with the grading and staging of the tumors.

Catestatin improves insulin sensitivity by attenuating endoplasmic reticulum stress: In vivo and in silico validation

Obesity is characterized by a state of chronic, unresolved inflammation in insulin-targeted tissues. Obesity-induced inflammation causes accumulation of proinflammatory macrophages in adipose tissue and liver. Proinflammatory cytokines released from tissue macrophages inhibits insulin sensitivity. Obesity also leads to inflammation-induced endoplasmic reticulum (ER) stress and insulin resistance. In this scenario, based on the data (specifically patterns) generated by our in vivo experiments on both diet-induced obese (DIO) and normal chow diet (NCD) mice, we developed an in silico state space model to integrate ER stress and insulin signaling pathways. Computational results successfully followed the experimental results for both DIO and NCD conditions. Chromogranin A (CgA) peptide catestatin (CST: hCgA352-372) improves obesity-induced hepatic insulin resistance by reducing inflammation and inhibiting proinflammatory macrophage infiltration. We reasoned that the anti-inflammatory effects of CST would alleviate ER stress. CST decreased obesity-induced ER dilation in hepatocytes and macrophages. On application of Proportional-Integral-Derivative (PID) controllers on the in silico model, we checked whether the reduction of phosphorylated PERK resulting in attenuation of ER stress, resembling CST effect, could enhance insulin sensitivity. The simulation results clearly pointed out that CST not only decreased ER stress but also enhanced insulin sensitivity in mammalian cells. In vivo experiment validated the simulation results by depicting that CST caused decrease in phosphorylation of UPR signaling molecules and increased phosphorylation of insulin signaling molecules. Besides simulation results predicted that enhancement of AKT phosphorylation helps in both overcoming ER stress and achieving insulin sensitivity. These effects of CST were verified in hepatocyte culture model.