RESUMEN
Introduction: The purpose of this study was to determine the possible differences in genetic polymorphisms and serum levels of chromogranin A (CgA), according to age and sex, in subjects with and without metabolic syndrome (MetS). Methodology: The genotyping and serum level of CgA and biochemical parameters were measured by the T-ARMS-PCR and PCR-RFLP and ELISA and spectrophotometer methods, respectively. Results: A comparison of males with and without MetS showed significantly lower high-density lipoprotein-cholesterol (HDL-C) levels than those of females.At ages 30-70 years, both sexes showed significant differences in triglycerides (TG), fasting blood sugar (FBS), CgA levels and waist circumference (WC) when compared to the two groups. Both sexes with MetS indicated significant differences in systolic blood pressure (SBP) at ages 40-70 years, while at ages 40-59 years, there was a significant difference in HDL-C level in males.There was a significant correlation between serum levels of FBS, TG, SBP and WC (in both sexes), and CgA in subjects with MetS. Significant correlation was found between HDL-C level and diastolic blood pressure (DBP), and CgA level in males and females, respectively. CgA genotype frequency (T-415C and C+87T polymorphisms) showed no significant differences between males and females with and without MetS, while there was only a significant difference in frequency of the genotypes T-415C when compared to males with and without MetS. Conclusion: The CgA appears to be strongly associated with MetS components in both sexes. Variation in CgA gene expression may affect the T-415C polymorphism in males. This may mean that the structure of CgA genetics differs in different ethnic groups. Differences in the serum level and expression of CgA gene may show valuable study results that it may be expected a relationship between these variables and the MetS.
Asunto(s)
Cromogranina A , Síndrome Metabólico , Humanos , Masculino , Femenino , Persona de Mediana Edad , Síndrome Metabólico/genética , Síndrome Metabólico/sangre , Adulto , Anciano , Cromogranina A/sangre , Cromogranina A/genética , Factores Sexuales , Factores de Edad , Polimorfismo Genético/genética , Presión Sanguínea/genética , Genotipo , Triglicéridos/sangre , HDL-Colesterol/sangre , Circunferencia de la Cintura/genéticaRESUMEN
OBJECTIVE: To explore the mechanism by which Qinghua decoction regulates neuroendocrine inflammation in chronic nonbacterial prostatitis (CNP) model rats and provide an experimental basis for clinical treatment. METHODS: The rats were randomly divided into six groups: normal control, model, Qianlie Tongyu capsule, low-dose Qinghua decoction, medium-dose Qinghua decoction, and high-dose Qinghua decoction group with six rats in each group. Rats in each group were sacrificed on the 29th day of treatment, and blood and prostate tissues were collected. Serum levels of tumor necrosis factor-alpha and interleukins 1-beta, 6, 8, and 10 (TNF-α and IL-1ß, -6, -8, and -10, respectively) were measured using enzyme-linked immunosorbent assay. The pathological changes in the rat prostate tissue in each group were observed under a light microscope. The expression levels of chromogranin A (CgA), nerve growth factor (NGF), and tyrosine kinase A (TrkA) were detected using reverse transcription quantitative polymerase chain reaction. Western blotting was used to detect protein expression of CgA, NGF, and TrkA. RESULTS: In the model group, the prostate capsule membrane and stroma were significantly dilated with more inflammatory cells infiltrating the stroma and perivessels. TNF-α, IL-1ß, -6, and -8, CgA, NGF, and TrkA levels increased, whereas the content of IL-10 decreased, which was statistically significant compared to that in the normal control group ( < 0.05). Prostate tissue cells in the high-dose group were neatly arranged with no obvious inflammatory cell infiltration. When compared with the model group, the high-dose Qinghua decoction group showed a significant improvement in these indices ( < 0.05). CONCLUSION: Qinghua decoction led to inhibition of pathological changes in the prostate tissue of rats with CNP, regulation of inflammatory cytokine expression, and inhibition in the expression of CgA, NGF, and TrkA. This mechanism may be primarily related to regulation of the CgA/NGF/TrkA signaling pathway mediated by various inflammatory factors.
Asunto(s)
Prostatitis , Masculino , Humanos , Ratas , Animales , Prostatitis/tratamiento farmacológico , Prostatitis/genética , Prostatitis/metabolismo , Proteínas Tirosina Quinasas/metabolismo , Cromogranina A/genética , Ratas Sprague-Dawley , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Factor de Crecimiento Nervioso/genética , Factor de Crecimiento Nervioso/metabolismo , Transducción de SeñalRESUMEN
Little is known about the adaptor protein FAM159B. Recently, FAM159B was shown to be particularly expressed in neuroendocrine cells and tissues, such as pancreatic islets and neuroendocrine cells of the bronchopulmonary and gastrointestinal tracts, as well as in different types of neuroendocrine tumours. To gain insights into possible interactions of FAM159B with other proteins and/or receptors, we analysed the co-expression of FAM159B and various neuroendocrine-specific markers in the cancer cell lines BON-1, PC-3, NCI-h82, OH-1, and A431 and also in human pancreatic tissues and pancreatic neuroendocrine tumours. The markers included prominent markers of neuroendocrine differentiation, such as chromogranin A (CgA), neuron-specific enolase (NSE), synaptophysin (SYP), insulinoma-associated protein 1 (INSM1), neural cell adhesion molecule 1 (NCAM1), serotonin (5-HT), somatostatin-14/28 (SST), and several receptors that are typically expressed by neuroendocrine cells, such as dopamine receptor 2 (D2R), somatostatin receptor (SSTR) 1, 2, 3, 4 and 5, and regulator of G-protein signalling 9 (RGS9). FAM159B was expressed evenly throughout the cytosol in all five cancer cell lines. Immunocytochemical and immunohistochemical analyses revealed co-expression of FAM159B with SYP, INSM1, RGS9, D2R, SSTR2, SSTR3, SSTR4, and SSTR5 and strong overlapping co-localisation with NSE. Double-labelling and co-immunoprecipitation Western blot analyses confirmed a direct association between FAM159B and NSE. These results suggest the involvement of FAM159B in several intracellular signalling pathways and a direct or indirect influence on diverse membrane proteins and receptors.
Asunto(s)
Células Neuroendocrinas , Tumores Neuroendocrinos , Neoplasias Pancreáticas , Humanos , Células Neuroendocrinas/metabolismo , Biomarcadores de Tumor/metabolismo , Tumores Neuroendocrinos/genética , Tumores Neuroendocrinos/metabolismo , Cromogranina A/genética , Cromogranina A/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Neoplasias Pancreáticas/patología , Proteínas Represoras/metabolismoRESUMEN
The aim of study was to assess genetic polymorphisms and serum level of chromogranin A (CgA) and its association with metabolic syndrome (MetS) components in the Fars ethnic group. Two hundred and forty-six people from Fars ethnic group participated in the study. The ATP III criteria were used to determine MetS components. The serum CgA level was measured by ELISA and the detection of the two regions were performed by TETRA ARMs-PCR and RFLPPCR methods. In results, the CC, CT, and TT genotypes of +87 C>T were 65%, 31.7%, and 3.2%, and were 74.8%, 25.2%, and 0% in subjects with and without MetS, respectively. The C and T alleles of +87 C>T were 81%, 19% and 79%, 21% in both groups, respectively. The TT, CT, and CC genotypes of -415 T>C were 76.4%, 21.1%, 2.4% and were 58.5%, 40.6% and 0.8% in subjects with and without MetS, respectively. The T and C alleles were 87% and 13% and 79% and 21% in both groups, respectively. There was correlation between serum level of fasting blood sugar (FBS) and CgA in subjects with MetS. We conclude that the increased CgA level in the subjects with MetS has a positive significant relationship with serum level of FBS. The most differences in CgA gene polymorphism were seen in -415 T>C genotype than that of +87C>T genotype when compared two groups. It may mean that subjects with MetS in the Fars ethnic group are more sensitive and greater risk for the development of MetS in the genotype of -415 T>C.
Asunto(s)
Síndrome Metabólico , Adenosina Trifosfato , Glucemia , Cromogranina A/genética , Etnicidad , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Síndrome Metabólico/genética , Polimorfismo de Nucleótido SimpleRESUMEN
The gut microbiota is in continuous interaction with the innermost layer of the gut, namely the epithelium. One of the various functions of the gut epithelium, is to keep the microbes at bay to avoid overstimulation of the underlying mucosa immune cells. To do so, the gut epithelia secrete a variety of antimicrobial peptides, such as chromogranin A (CgA) peptide catestatin (CST: hCgA352-372). As a defense mechanism, gut microbes have evolved antimicrobial resistance mechanisms to counteract the killing effect of the secreted peptides. To this end, we treated wild-type mice and CST knockout (CST-KO) mice (where only the 63 nucleotides encoding CST have been deleted) with CST for 15 consecutive days. CST treatment was associated with a shift in the diversity and composition of the microbiota in the CST-KO mice. This effect was less prominent in WT mice. Levels of the microbiota-produced short-chain fatty acids, in particular, butyrate and acetate were significantly increased in CST-treated CST-KO mice but not the WT group. Both CST-treated CST-KO and WT mice showed a significant increase in microbiota-harboring phosphoethanolamine transferase-encoding genes, which facilitate their antimicrobial resistance. Finally, we show that CST was degraded by Escherichia coli via an omptin-protease and that the abundance of this gene was significantly higher in metagenomic datasets collected from patients with Crohn's disease but not with ulcerative colitis. Overall, this study illustrates how the endogenous antimicrobial peptide, CST, shapes the microbiota composition in the gut and primes further research to uncover the role of bacterial resistance to CST in disease states such as inflammatory bowel disease.
Asunto(s)
Antiinfecciosos , Microbioma Gastrointestinal , Animales , Cromogranina A/genética , Cromogranina A/metabolismo , Cromogranina A/farmacología , Ratones , Ratones Noqueados , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/metabolismo , PéptidosRESUMEN
Pancreastatin (PST), a chromogranin A-derived potent physiological dysglycemic peptide, regulates glucose/insulin homeostasis. We have identified a nonsynonymous functional PST variant (p.Gly297Ser; rs9658664) that occurs in a large section of human populations. Association analysis of this single nucleotide polymorphism with cardiovascular/metabolic disease states in Indian populations (n = 4,300 subjects) displays elevated plasma glucose, glycosylated hemoglobin, diastolic blood pressure, and catecholamines in Gly/Ser subjects as compared with wild-type individuals (Gly/Gly). Consistently, the 297Ser allele confers an increased risk (â¼1.3-1.6-fold) for type 2 diabetes/hypertension/coronary artery disease/metabolic syndrome. In corroboration, the variant peptide (PST-297S) displays gain-of-potency in several cellular events relevant for cardiometabolic disorders (e.g., increased expression of gluconeogenic genes, increased catecholamine secretion, and greater inhibition of insulin-stimulated glucose uptake) than the wild-type peptide. Computational docking analysis and molecular dynamics simulations show higher affinity binding of PST-297S peptide with glucose-regulated protein 78 (GRP78) and insulin receptor than the wild-type peptide, providing a mechanistic basis for the enhanced activity of the variant peptide. In vitro binding assays validate these in silico predictions of PST peptides binding to GRP78 and insulin receptor. In conclusion, the PST 297Ser allele influences cardiovascular/metabolic phenotypes and emerges as a novel risk factor for type 2 diabetes/hypertension/coronary artery disease in human populations.
Asunto(s)
Enfermedades Cardiovasculares/genética , Cromogranina A/genética , Predisposición Genética a la Enfermedad/genética , Enfermedades Metabólicas/genética , Secuencia de Aminoácidos , Animales , Catecolaminas/sangre , Línea Celular , Línea Celular Tumoral , Cromogranina A/química , Cromogranina A/metabolismo , Enfermedad de la Arteria Coronaria/genética , Diabetes Mellitus Tipo 2/genética , Chaperón BiP del Retículo Endoplásmico/metabolismo , Estudios de Asociación Genética/métodos , Células Hep G2 , Humanos , Hipertensión/genética , India , Péptidos/química , Péptidos/genética , Péptidos/metabolismo , Polimorfismo de Nucleótido Simple/genética , Ratas , Receptor de Insulina/metabolismoRESUMEN
Vasostatin 1 (VS-1) plays an important role in the regulation of various tissue injury and repair processes, but its role in aortic aneurysm remains unclear. The plasmid-like nanoparticles containing the vasostatin-1 gene Pul-PGEA-pCas-sgVs-1 were constructed, and their guarantee, safety, hemolysis, and particle size were analyzed. Eighty-four eight-week-old male ApoE-mice were randomly divided into blank group (without any treatment), model group (Ang II aortic aneurysm model + tail injection of PBS), control group (modeling + tail injection of Pul-PGEA-pCas9), and experimental group (modeling + tail injection of Pul-PGEA-pCas-sgVs-1), with 21 rats in each group. The incidence, mortality, and maximum diameter of abdominal aortic aneurysm (AAA) and the contents of high sensitivity C-reactive protein (HS-CRP), soluble intercellular adhesion molecule-1 (ICAM-1), soluble vascular cell adhesion molecule-1 (VCAM-1), and TNF-a in serum were compared in different groups of mice. The results showed that Pul-PGEA-pCas-sgVs-1 had good biosafety and transfection ability. The maximum diameter of abdominal aorta, incidence of abdominal aortic aneurysm, mortality, and the expression levels of HS-CRP, ICAM-1, VCAM-1, and TNF-a in the experimental group were lower than those in the model group (P< 0.05). These results indicated that the plasmid-like nanoparticles Pul-PGEA-pCas-sgVs-1 can inhibit the development of aorta by down-regulating the expression of inflammatory factors, which played a good protective role on the aorta.
Asunto(s)
Aneurisma de la Aorta Abdominal , Cromogranina A , Regulación de la Expresión Génica , Nanopartículas , Fragmentos de Péptidos , Plásmidos , Animales , Aneurisma de la Aorta Abdominal/genética , Aneurisma de la Aorta Abdominal/metabolismo , Aneurisma de la Aorta Abdominal/prevención & control , Cromogranina A/biosíntesis , Cromogranina A/genética , Modelos Animales de Enfermedad , Masculino , Ratones , Ratones Noqueados para ApoE , Nanopartículas/química , Nanopartículas/uso terapéutico , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/genética , Plásmidos/química , Plásmidos/genética , Plásmidos/farmacologíaRESUMEN
OBJECTIVE: Because of their rarity, it is not known whether isocitrate dehydrogenase 2 (IDH2) mutations are related to the occurrence of olfactory neuroblastoma (ONB). We investigated the relationships between IDH2 mutations, clinicopathological parameters, and the prognosis for ONB to establish a molecular classification using IDH2 mutations. METHODS: An 82-patient cohort was retrospectively screened using immunohistochemistry with a mutation-specific IDH2 antibody and real-time polymerase chain reactions for IDH2 mutations. We also immunohistochemically determined the expression of chromogranin A, synaptophysin, neuron-specific enolase, CD56, S100, and Ki-67. RESULTS: The 2 methods used for the detection of IDH2 mutations had high consistency. Mutation of IDH2 detected by real-time polymerase chain reaction correlated with higher Kadish stage, Hyams grade, and Ki-67 proliferation index. Mutation of IDH2 correlated negatively with the expression of chromogranin A, synaptophysin, CD56, and S100. Kaplan-Meier analysis showed that an IDH2 mutation, a high Hyams grade, and high Ki-67 proliferation index were associated with poor overall survival. The Hyams grade and IDH2 mutation were independent prognostic factors on multivariable analysis. CONCLUSIONS: Immunohistochemistry was a reliable method to assess the mutation status of IDH2. Tumors with IDH2 mutations represented a distinct subset with aggressive behavior and conferred a poor prognosis. The gene status of IDH2 could be a major molecular classification criterion in ONB.
Asunto(s)
Estesioneuroblastoma Olfatorio , Neoplasias Nasales , Cromogranina A/genética , Estesioneuroblastoma Olfatorio/genética , Estesioneuroblastoma Olfatorio/patología , Humanos , Isocitrato Deshidrogenasa/genética , Antígeno Ki-67/genética , Mutación/genética , Cavidad Nasal/patología , Neoplasias Nasales/genética , Neoplasias Nasales/patología , Pronóstico , Estudios Retrospectivos , SinaptofisinaRESUMEN
BACKGROUND: Although preeclampsia has long been recognized as a condition affecting late pregnancy, little is known of its pathogenesis or treatment. The placenta releases a number of hormones and molecules that influence the course of pregnancy, one of which is chromogranin A, a soluble protein secreted mainly from the chromaffin cells of the adrenal medulla. Its role in pregnancy and pregnancy-related disorders remains unclear. Therefore, the main aim of the proposed study is to determine whether chromogranin A is related with the occurrence of preeclampsia. METHODS: Placental samples were collected from 102 preeclamptic patients and 103 healthy controls, and Chromogranin A gene (CHGA) expression was measured using real-time RT-PCR, The RT-PCR results were verified on the protein level using ELISA. The normal distribution of the data was tested using the Shapiro-Wilk test. The clinical and personal characteristics of the groups were compared using the Student's t-test for normally-distributed data, and the χ2 test for categorical variables. The Mann-Whitney U test was used for non-normally distributed data. As the log- transformation was not suitable for the given outcomes, the Box- Cox Transformation was used to normalize data from ELISA tests and CHGA expression. Values of P < .05 were considered statistically significant. RESULTS: Chromogranin A gene expression was found to be significantly higher in the study group than in controls. Protein analyses showed that although the CgA concentration in placental samples did not differ significantly, the catestatin (CST) level was significantly lower in samples obtained from women with preeclampsia, according to the controls. CONCLUSIONS FOR PRACTICE: This study for the first time reveals that chromogranin A gene expression level is associated with preeclampsia. Moreover, the depletion in catestatin level, which plays a protective role in hypertension development, might be a marker of developing preeclampsia. Further studies may unravel role of Chromogranin A in the discussed disease.
Asunto(s)
Cromogranina A/metabolismo , Fragmentos de Péptidos/metabolismo , Placenta/metabolismo , Preeclampsia/metabolismo , Biomarcadores/metabolismo , Estudios de Casos y Controles , Cromogranina A/genética , Femenino , Expresión Génica , Humanos , Fragmentos de Péptidos/genética , Preeclampsia/genética , EmbarazoRESUMEN
Recognition of ß-cell antigens by autoreactive T cells is a critical step in the initiation of autoimmune type1 diabetes. A complete protection from diabetes development in NOD mice harboring a point mutation in the insulin B-chain 9-23 epitope points to a dominant role of insulin in diabetogenesis. Generation of NOD mice lacking the chromogranin A protein (NOD.ChgA-/-) completely nullified the autoreactivity of the BDC2.5 T cell and conferred protection from diabetes onset. These results raised the issue concerning the dominant antigen that drives the autoimmune process. Here we revisited the NOD.ChgA-/- mice and found that their lack of diabetes development may not be solely explained by the absence of chromogranin A reactivity. NOD.ChgA-/- mice displayed reduced presentation of insulin peptides in the islets and periphery, which corresponded to impaired T-cell priming. Diabetes development in these mice was restored by antibody treatment targeting regulatory T cells or inhibiting transforming growth factor-ß and programmed death-1 pathways. Therefore, the global deficiency of chromogranin A impairs recognition of the major diabetogenic antigen insulin, leading to broadly impaired autoimmune responses controlled by multiple regulatory mechanisms.
Asunto(s)
Autoinmunidad/genética , Cromogranina A/genética , Diabetes Mellitus Tipo 1/genética , Animales , Presentación de Antígeno/genética , Autoantígenos/inmunología , Autoantígenos/metabolismo , Citoprotección/genética , Citoprotección/inmunología , Diabetes Mellitus Tipo 1/prevención & control , Epítopos de Linfocito T/inmunología , Femenino , Islotes Pancreáticos/inmunología , Islotes Pancreáticos/metabolismo , Ratones , Ratones Endogámicos NOD , Ratones NoqueadosRESUMEN
[Figure: see text].
Asunto(s)
Cardiotónicos/farmacología , Cromogranina A/genética , Hipertensión/tratamiento farmacológico , Hipertrofia Ventricular Izquierda/genética , Macrófagos/efectos de los fármacos , Fragmentos de Péptidos/farmacología , Animales , Cardiotónicos/uso terapéutico , Cromogranina A/metabolismo , Cromogranina A/farmacología , Cromogranina A/uso terapéutico , Hipertensión/metabolismo , Hipertensión/patología , Hipertrofia Ventricular Izquierda/metabolismo , Hipertrofia Ventricular Izquierda/patología , Terapia de Inmunosupresión , Macrófagos/metabolismo , Macrófagos/patología , Masculino , Ratones , Ratones Noqueados , Fragmentos de Péptidos/uso terapéuticoRESUMEN
Adenocarcinoma admixed with neuroendocrine carcinoma of the uterine cervix is a rare malignancy with a poor prognosis. In the literature, there are few reported cases. Herein, we report a case of a 56-year-old Turkish woman with cervical adenocarcinoma admixed with small cell neuroendocrine carcinoma. Histological examination of endocervical curettage specimens revealed a tumor composed of almost equal areas of small cell neuroendocrine carcinoma and adenocarcinoma. Neuroendocrine differentiation was confirmed by immunohistochemistry for chromogranin-A, synaptophysin, and CD 56. After the adenocarcinoma and small cell neuroendocrine carcinoma association was detected in the curettage material, both cervicovaginal smear and then total abdominal hysterectomy and bilateral salpingo-oophorectomy resection material of the patient were submitted to our pathology department. Histological features of both curettage and resection material were determined by immunohistochemical studies.
Asunto(s)
Adenocarcinoma/diagnóstico , Carcinoma Neuroendocrino/diagnóstico , Neoplasias del Cuello Uterino/diagnóstico , Adenocarcinoma/clasificación , Adenocarcinoma/cirugía , Antígeno CD56/genética , Carcinoma Neuroendocrino/cirugía , Cuello del Útero/patología , Cromogranina A/genética , Femenino , Humanos , Histerectomía , Inmunohistoquímica , Persona de Mediana Edad , Sinaptofisina/genética , Neoplasias del Cuello Uterino/clasificación , Neoplasias del Cuello Uterino/cirugíaRESUMEN
Background: Ulcerative colitis (UC) is characterized by altered chromogranin-A (CHGA), alternatively activated macrophages (M2) and intestinal epithelial cells (IECs). We previously demonstrated that CHGA is implicated in colitis progression by regulating the macrophages. Here, we investigated the interplay between CHGA, M2, tight junctions (TJ) and IECs in an inflammatory environment. Methods: Correlations between CHGA mRNA expression of and TJ proteins mRNA expressions of (Occludin [OCLN], zonula occludens-1 [ZO1], Claudin-1 [CLDN1]), epithelial associated cytokines (interleukin [IL]-8, IL-18), and collagen (COL1A2) were determined in human colonic mucosal biopsies isolated from active UC and healthy patients. Acute UC-like colitis (5% dextran sulphate sodium [DSS], five days) was induced in Chga-C57BL/6-deficient (Chga-/-) and wild type (Chga+/+) mice. Col1a2 TJ proteins, Il-18 mRNA expression and collagen deposition were determined in whole colonic sections. Naïve Chga-/- and Chga+/+ peritoneal macrophages were isolated and exposed six hours to IL-4/IL-13 (20 ng/mL) to promote M2 and generate M2-conditioned supernatant. Caco-2 epithelial cells were cultured in the presence of Chga-/- and Chga+/+ non- or M2-conditioned supernatant for 24 h then exposed to 5% DSS for 24 h, and their functional properties were assessed. Results: In humans, CHGA mRNA correlated positively with COL1A2, IL-8 and IL-18, and negatively with TJ proteins mRNA markers. In the experimental model, the deletion of Chga reduced IL-18 mRNA and its release, COL1A2 mRNA and colonic collagen deposition, and maintained colonic TJ proteins. Chga-/- M2-conditioned supernatant protected caco-2 cells from DSS and oxidative stress injuries by improving caco-2 cells functions (proliferation, viability, wound healing) and by decreasing the release of IL-8 and IL-18 and by maintaining the levels of TJ proteins, and when compared with Chga+/+ M2-conditioned supernatant. Conclusions: CHGA contributes to the development of intestinal inflammation through the regulation of M2 and epithelial cells. Targeting CHGA may lead to novel biomarkers and therapeutic strategies in UC.
Asunto(s)
Cromogranina A/genética , Colitis Ulcerosa/inmunología , Citocinas/genética , Macrófagos/inmunología , Proteínas de Uniones Estrechas/genética , Animales , Células CACO-2 , Estudios de Casos y Controles , Células Cultivadas , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/genética , Sulfato de Dextran/efectos adversos , Modelos Animales de Enfermedad , Técnicas de Inactivación de Genes , Humanos , Interleucina-18/genética , Interleucina-8/genética , Activación de Macrófagos , Ratones , Ratones Endogámicos C57BLRESUMEN
Neuroendocrine tumors (NETs) are often diagnosed from the metastases of an unknown primary tumor. Specific immunohistochemical (IHC) markers indicating the location of a primary tumor are needed. The proprotein convertase subtilisin/kexin type 2 (PCSK2) is found in normal neural and neuroendocrine cells, and known to express in NETs. We investigated the tissue microarray (TMA) of 86 primary tumors from 13 different organs and 9 metastatic NETs, including primary tumor-metastasis pairs, for PCSK2 expression with polymer-based IHC. PCSK2 was strongly positive in all small intestine and appendiceal NETs, the so-called midgut NETs, in most pheochromocytomas and paragangliomas, and in some of the typical and atypical pulmonary carcinoid tumors. NETs showing strong positivity were re-evaluated in larger tumor cohorts confirming the primary observation. In the metastases, the expression of PCSK2 mirrored that of the corresponding primary tumors. We found negative or weak staining in NETs from the thymus, gastric mucosa, pancreas, rectum, thyroid, and parathyroid. PCSK2 expression did not correlate with Ki-67 in well-differentiated NETs. Our data suggest that PCSK2 positivity can indicate the location of the primary tumor. Thus, PCSK2 could function in the IHC panel determined from screening metastatic NET biopsies of unknown primary origins.
Asunto(s)
Neoplasias de las Glándulas Suprarrenales/genética , Carcinoma Neuroendocrino/genética , Neoplasias Gastrointestinales/genética , Neoplasias Pulmonares/genética , Tumores Neuroendocrinos/genética , Paraganglioma/genética , Feocromocitoma/genética , Proproteína Convertasa 2/genética , Adenocarcinoma/diagnóstico , Adenocarcinoma/genética , Adenocarcinoma/patología , Adenocarcinoma/cirugía , Neoplasias de las Glándulas Suprarrenales/diagnóstico , Neoplasias de las Glándulas Suprarrenales/patología , Neoplasias de las Glándulas Suprarrenales/cirugía , Biomarcadores de Tumor/genética , Carcinoma Neuroendocrino/diagnóstico , Carcinoma Neuroendocrino/patología , Carcinoma Neuroendocrino/cirugía , Cromogranina A/genética , Femenino , Neoplasias Gastrointestinales/diagnóstico , Neoplasias Gastrointestinales/patología , Neoplasias Gastrointestinales/cirugía , Expresión Génica , Humanos , Inmunohistoquímica , Antígeno Ki-67/genética , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/cirugía , Metástasis Linfática , Masculino , Estadificación de Neoplasias , Tumores Neuroendocrinos/diagnóstico , Tumores Neuroendocrinos/patología , Tumores Neuroendocrinos/cirugía , Paraganglioma/diagnóstico , Paraganglioma/patología , Paraganglioma/cirugía , Feocromocitoma/diagnóstico , Feocromocitoma/patología , Feocromocitoma/cirugía , Neoplasias de la Tiroides/diagnóstico , Neoplasias de la Tiroides/genética , Neoplasias de la Tiroides/patología , Neoplasias de la Tiroides/cirugíaRESUMEN
Prion diseases affect both animals and humans. Research in the natural animal model of the disease could help in the understanding of neuropathological mechanisms and in the development of biomarkers for human pathologies. For this purpose, we studied the expression of 10 genes involved in prion propagation in vitro in the central nervous system of scrapie-infected sheep. Dysregulated genes (BAMBI and CHGA) were further analysed in a transgenic murine model (Tg338) of scrapie, and their protein distribution was determined using immunohistochemistry and Western blot. Their potential as biomarkers was finally assessed using enzyme-linked immunosorbent assay (ELISA) in cerebrospinal fluid (CSF) of scrapie sheep and Creutzfeldt-Jakob disease (CJD) patients. Protein BAMBI was upregulated in highly affected brain areas and CHGA was overexpressed along the brain in both models. Moreover, BAMBI and CHGA immunostaining scores strongly correlated with spongiosis and microgliosis in mice. Finally, levels of BAMBI were significantly higher in the CSF of clinical sheep and CJD patients. In addition to their potential as biomarkers, our work confirms the role of BAMBI and CHGA in prion neuropathology in vivo, but besides prion replication, they seem to be involved in the characteristic neuroinflammatory response associated to prion infection.
Asunto(s)
Cromogranina A/líquido cefalorraquídeo , Síndrome de Creutzfeldt-Jakob/líquido cefalorraquídeo , Proteínas de la Membrana/líquido cefalorraquídeo , Scrapie/líquido cefalorraquídeo , Animales , Biomarcadores/líquido cefalorraquídeo , Biomarcadores/metabolismo , Encéfalo/metabolismo , Encéfalo/patología , Cromogranina A/genética , Cromogranina A/metabolismo , Síndrome de Creutzfeldt-Jakob/patología , Humanos , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Ratones , Scrapie/patología , OvinosAsunto(s)
Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas de Esófago/genética , Neoplasias de Cabeza y Cuello/genética , Neoplasias Pulmonares/genética , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Actinas/genética , Adulto , Anciano , Biomarcadores de Tumor/genética , Cromogranina A/genética , Conexina 43/genética , Neoplasias Esofágicas/patología , Carcinoma de Células Escamosas de Esófago/patología , Perfilación de la Expresión Génica/métodos , Neoplasias de Cabeza y Cuello/patología , Humanos , Neoplasias Pulmonares/patología , Masculino , Persona de Mediana Edad , Neoplasias Primarias Múltiples/genética , Neoplasias Primarias Múltiples/patología , Estudios Retrospectivos , Carcinoma de Células Escamosas de Cabeza y Cuello/patologíaRESUMEN
BACKGROUND: Chromogranin A (CHGA) is an index granin protein critical for biogenesis and exocytotic release of catecholamine storage granules. It is elevated in plasma of patients with sympathetic over-activity and kidney dysfunction. Several CHGA polymorphisms are associated with hypertensive kidney disease. Previously, we unraveled the molecular mechanism by which CHGA expression is regulated in African Americans carrying a genetic variation associated with hypertensive chronic kidney disease (CKD). METHOD: Experimental CKD mouse model were created by 5/6th nephrectomy (Npx) using wild-type and Chga-/- knockout mouse strains to delineate the role of CHGA in CKD. RESULT: Wild-type-Npx mice expressing Chga developed exacerbated azotemia and fibrosis as compared with their knockout-Npx counterparts. Gene expression profiling revealed downregulation of mitochondrial respiratory complexes genes consistent with maladaptive mitochondria in wild-type-Npx mice, contrasted to knockout-Npx. In healthy individuals, an inverse relationship between circulating CHGA levels and glomerular function was observed. In vitro, mesangial cells treated with CHGA-triggered nitric oxide release by a signaling mechanism involving scavenger receptor SR-A. The CHGA-treated and untreated mesangial cells displayed differential expression of cytokine, chemokine, complement, acute phase inflammatory and apoptotic pathway genes. Thus, build-up of plasma CHGA because of kidney injury served as an insult to the mesangial cells resulting in expression of genes promoting inflammation, fibrosis, and progression of CKD. CONCLUSION: These findings improve understanding of the role of elevated CHGA in the progression of CKD and reveal novel pathways that could be exploited for therapeutic strategies in hypertensive kidney disease.
Asunto(s)
Cromogranina A , Hipertensión Renal , Nefritis , Animales , Cromogranina A/genética , Cromogranina A/metabolismo , Hipertensión Renal/genética , Hipertensión Renal/metabolismo , Hipertensión Renal/patología , Ratones , Ratones Noqueados , Nefritis/genética , Nefritis/metabolismo , Nefritis/patologíaRESUMEN
Colon cancer is one of the major causes of cancer death worldwide. The five-year survival rate for the early-stage patients is more than 90%, and only around 10% for the later stages. Moreover, half of the colon cancer patients have been clinically diagnosed at the later stages. It is; therefore, of importance to enhance the ability for the early diagnosis of colon cancer. Taking advantages from our previous studies, there are several potential biomarkers which have been associated with the early diagnosis of the colon cancer. In order to investigate these early diagnostic biomarkers for colon cancer, human chromogranin-A (CHGA) was further analyzed among the most powerful diagnostic biomarkers. In this study, we used a logistic regression-based meta-analysis to clarify associations of CHGA expression with colon cancer diagnosis. Both healthy populations and the normal mucosa from the colon cancer patients were selected as the double normal controls. The results showed decreased expression of CHGA in the early stages of colon cancer as compared to the normal controls. The decline of CHGA expression in the early stages of colon cancer is probably a new diagnostic biomarker for colon cancer diagnosis with high predicting possibility and verification performance. We have also compared the diagnostic powers of CHGA expression with the typical oncogene KRAS, classic tumor suppressor TP53, and well-known cellular proliferation index MKI67, and the CHGA showed stronger ability to predict early diagnosis for colon cancer than these other cancer biomarkers. In the protein-protein interaction (PPI) network, CHGA was revealed to share some common pathways with KRAS and TP53. CHGA might be considered as a novel, promising, and powerful biomarker for early diagnosis of colon cancer.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Cromogranina A/metabolismo , Neoplasias del Colon/metabolismo , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/normas , Cromogranina A/genética , Cromogranina A/normas , Neoplasias del Colon/genética , Neoplasias del Colon/patología , Redes Reguladoras de Genes , Humanos , Metaanálisis como Asunto , Mapas de Interacción de Proteínas , Proteínas Proto-Oncogénicas p21(ras)/genética , Proteínas Proto-Oncogénicas p21(ras)/metabolismo , Sensibilidad y Especificidad , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
CD4 T cells play a critical role in promoting the development of autoimmunity in type 1 diabetes. The diabetogenic CD4 T cell clone BDC-2.5, originally isolated from a NOD mouse, has been widely used to study the contribution of autoreactive CD4 T cells and relevant Ags to autoimmune diabetes. Recent work from our laboratory has shown that the Ag for BDC-2.5 T cells is a hybrid insulin peptide (2.5HIP) consisting of an insulin C-peptide fragment fused to a peptide from chromogranin A (ChgA) and that endogenous 2.5HIP-reactive T cells are major contributors to autoimmune pathology in NOD mice. The objective of this study was to determine if poly(lactide-co-glycolide) (PLG) nanoparticles (NPs) loaded with the 2.5HIP Ag (2.5HIP-coupled PLG NPs) can tolerize BDC-2.5 T cells. Infusion of 2.5HIP-coupled PLG NPs was found to prevent diabetes in an adoptive transfer model by impairing the ability of BDC-2.5 T cells to produce proinflammatory cytokines through induction of anergy, leading to an increase in the ratio of Foxp3+ regulatory T cells to IFN-γ+ effector T cells. To our knowledge, this work is the first to use a hybrid insulin peptide, or any neoepitope, to re-educate diabetogenic T cells and may have significant implications for the development of an Ag-specific therapy for type 1 diabetes patients.
Asunto(s)
Cromogranina A/metabolismo , Diabetes Mellitus Tipo 1/terapia , Inmunoterapia/métodos , Insulina/metabolismo , Nanopartículas/uso terapéutico , Péptidos/metabolismo , Proteínas Recombinantes de Fusión/uso terapéutico , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Reguladores/inmunología , Animales , Animales Modificados Genéticamente , Células Cultivadas , Cromogranina A/genética , Diabetes Mellitus Tipo 1/inmunología , Modelos Animales de Enfermedad , Factores de Transcripción Forkhead/metabolismo , Humanos , Tolerancia Inmunológica , Insulina/genética , Interferón gamma/metabolismo , Ratones , Ratones Endogámicos NOD , Nanopartículas/metabolismo , Péptidos/genética , Receptores de Antígenos de Linfocitos T/genética , Proteínas Recombinantes de Fusión/genéticaRESUMEN
Medullary thyroid carcinoma (MTC) has been shown to express Prospero homeobox protein 1 (Prox1), a transcription factor whose expression is altered in a variety of human cancers. We conducted a retrospective study on a series of 32 patients with MTC to test the correlation of Prox1 expression in MTC with clinicopathological features and to evaluate its prognostic significance. Correlation of Prox1 immunohistochemical expression with tumor size, proliferative index (Ki67), and calcitonin and CEA serum levels prior to surgery was tested for significant correlations. The difference in Prox1 and Ki67 immunohistochemical expression according to the immunohistochemical staining intensity of CEA, chromogranin A, and calcitonin was tested using the Kruskal-Wallis H test and linear regression analysis. The prognostic value of Prox1 and Ki67 for our patient cohort was assessed by Kaplan-Meier log rank survival analysis. We demonstrated a positive correlation between Prox1 expression and Ki67 index. Prox1 also showed significant difference in expression according to chromogranin A and calcitonin immunohistochemical expression, with higher Prox1 expression in tumors with stronger chromogranin A or calcitonin staining. Prox1 expression did not correlate with PFS or OS based on Kaplan-Meier log rank survival analysis. In conclusion, Prox1 expression in MTC is positively correlated with Ki67 and with the immunohistochemical expression of chromogranin A and calcitonin. However, the present study does not support a role for Prox1 in MTC prognosis.