Elżbieta Wrotkowska

Chromogranin A – unspecific neuroendocrine marker. Clinical utility and potential diagnostic pitfalls

Chromogranin A, despite a number of limitations, is still the most valuable marker of neuroendocrine tumors (NETs). Granins belong to the family of acidic proteins that constitute a major component of secretory granules of various endocrine and neuroendocrine cells, which are components of both the classical endocrine glands and the diffuse neuroendocrine system. These cells are a potential source of transformation into neuroendocrine tumors. The awareness of potential causes influencing the false results of its concentrations simplifies diagnosis and treatment. One of the disadvantages of this marker is its non-specificity and the existence of a number of pathological processes leading to an increase in its concentration, which often results in confusion and diagnostic difficulties. The molecular structure is characterized by a number of sites susceptible to the proteolytic activity of enzymes, resulting in the formation of a number of biologically active peptides. Presumably they act as precursors of active proteins. Chromogranin expression correlates with the amount of secretory vesicles in neuroendocrine cells. The peptide chain during biochemical changes becomes a precursor of biologically active proteins with a wide range of activities. There are a number of commercially available kits for the determination of chromogranin A, which differ in methodology. We present the evaluation of chromogranin A as a marker of neuroendocrine tumors in clinical practice and the possible factors that may affect the outcome of its concentration.

Survivin Delta Ex3 Overexpression in Thyroid Malignancies

Context Thyroid cancer incidence has increased significantly during the past decades and is the most common type of endocrine malignancy. Many factors in thyroid cancers were studied as independent predictors of a poor prognosis. Objective The objective of the study was to evaluate survivin expression – BIRC5 and its splice variants: survivin delta Ex3 and survivin 2B in benign and malignant thyroid nodules. Design Thyroid tissues samples from a group of 50 patients consisting of: 29 patients with thyroid cancers (including medullary, papillary, follicular and undifferentiated types), as well as from 21 patients with non-cancerous thyroid tissues (including: 11 benign thyroid lesions and 10 healthy thyroid samples). Main Outcome Measures The analysis of the survivin gene expression and evaluation of the level of splice variants were performed using quantitative RT-PCR. Results A statistically significant higher level of expression of survivin gene – BIRC5 was detected in thyroid malignant nodules, when compared with benign lesions and healthy thyroid samples. Moreover, the comparison of survivin relative expression in different staged tumors (pT1, pT3, and pT4) revealed a much higher amount of BIRC5 transcripts in tumor tissues of pT3/pT4. The comparison of survivin expression between benign thyroid nodules and healthy thyroid did not reveal significant differences. Importantly, high expression rate of the survivin delta Ex3 splice variant characterized thyroid carcinomas. Conclusion The results suggest that survivin, especially survivin delta Ex3 splice variant being overexpress, is a characteristic feature of thyroid malignancy.

Individual plasma ghrelin changes in the same patients in hyperthyroid, hypothyroid and euthyroid state

Ghrelin is a multifunctional peptide of widespread expression. Since it has been shown to influence energy homeostatis, its potential role in thyroid dysfunction may have clinical significance. In this study, plasma ghrelin changes have been analyzed in the same patients in three different thyroid states for the first time. The study group consisted of 16 patients who had been diagnosed with hyperthyroidism, were treated with radioiodine, developed hypothyroidism after treatment, and finally became euthyroid on l-thyroxine substitution. In the initial state of hyperthyroidism plasma ghrelin levels correlated negatively with fT3 and fT4. In hypothyroidism ghrelin concentration increased significantly (p<0.05). Although the mean value of plasma ghrelin tended to decrease in the euthyroid state, the individual difference between hypothyroidism and euthyroidism was not significant. Plasma ghrelin in euthyroidism was still significantly higher than in hyperthyroidism (p<0.05), and correlated positively with ghrelin levels in hyperthyroidism and hypothyroidism. In our opinion, plasma ghrelin fluctuations may reflect metabolic changes in patients with thyroid dysfunction. Moreover, it cannot be excluded that in thyroid disorders ghrelin acts as a compensatory factor, helping to balance metabolic disturbances.

Time-dependent irisin concentration changes in patients affected by overt hypothyroidism.

INTRODUCTION Irisin, a cleaved and secreted part of the transmembrane protein FNDC5, is a recently discovered adipo-myokine that is said to have a significant influence on body metabolism. Changes in thyrometabolic state may also alter the serum irisin level. Since already reported data are not fully consistent, the aim of the present research is to evaluate the time-dependent changes in serum irisin level in patients affected by overt hypothyroidism. MATERIAL AND METHODS The study involved 36 subjects - two groups of 12 patients with long-lasting (AITD) and short-term (TC) overt hypothyroidism, and a control group (CG) of 12 subjects, matched for age and gender. Serum irisin level, thyrometabolic state, creatine kinase (CK - muscle damage marker), glucose, and insulin concentration were assessed and compared between groups. RESULTS The irisin level was significantly lower in AITD than in TC and CG (p = 0.02; p < 0.01; respectively) patients, with no statistical difference between TC and CG (p > 0.05). There was no significant difference between free triiodothyronine and free thyroxine levels in AITD and TC patients (p > 0.05). CK concentration was significantly higher in AITD than in CG patients (p < 0.01) with no difference between AITD and TC patients (p > 0.05) as well as TC and CG patients (p > 0.05). Additionally, the CK level negatively correlated with the irisin level (r = -0.58; p < 0.01). CONCLUSIONS In conclusion, the irisin concentration changes during thyroid function impairment may be time-dependent. Patients with prolonged hypothyroidism have lower irisin levels that those with short-term disorder. (Endokrynol Pol 2016; 67 (5): 476-480).

Familial syndromes associated with neuroendocrine tumours.

Neuroendocrine tumours may be associated with familial syndromes. At least eight inherited syndromes predisposing to endocrine neoplasia have been identified. Two of these are considered to be major factors predisposing to benign and malignant endocrine tumours, designated multiple endocrine neoplasia type 1 and type 2 (MEN1 and MEN2). Five other autosomal dominant diseases show more heterogeneous clinical patterns, such as the Carney complex, hyperparathyroidism-jaw tumour syndrome, Von Hippel-Lindau syndrome (VHL), neurofibromatosis type 1 (NF1) and tuberous sclerosis. The molecular and cellular interactions underlying the development of most endocrine cells and related organs represent one of the more complex pathways not yet to be deciphered. Almost all endocrine cells are derived from the endoderm and neuroectoderm. It is suggested that within the first few weeks of human development there are complex interactions between, firstly, the major genes involved in the initiation of progenitor-cell differentiation, secondly, factors secreted by the surrounding mesenchyme, and thirdly, a series of genes controlling cell differentiation, proliferation and migration. Together these represent a formula for the harmonious development of endocrine glands and tissue.

Management of the hormonal syndrome of neuroendocrine tumors

Gastroenteropancreatic neuroendocrine tumors (GEP/NET) are unusual and rare neoplasms that present many clinical challenges. They characteristically synthesize store and secrete a variety of peptides and neuroamines which can lead to the development of distinct clinical syndrome, however many are clinically silent until late presentation with mass effects. Management strategies include surgery cure and cytoreduction with the use of somatostatin analogues. Somatostatin have a broad range of biological actions that include inhibition of exocrine and endocrine secretions, gut motility, cell proliferation, cell survival and angiogenesis. Five somatostatin receptors (SSTR1-SSTR5) have been cloned and characterized. Somatostatin analogues include octreotide and lanreotide are effective medical tools in the treatment and present selectivity for SSTR2 and SSTR5. During treatment is seen disapperance of flushing, normalization of bowel movements and reduction of serotonin and 5-hydroxyindole acetic acid (5-HIAA) secretion. Telotristat represents a novel approach by specifically inhibiting serotonin synthesis and as such, is a promising potential new treatment for patients with carcinoid syndrome. To pancreatic functionig neuroendocrine tumors belongs insulinoma, gastrinoma, glucagonoma and VIP-oma. Medical management in patients with insulinoma include diazoxide which suppresses insulin release. Also mTOR inhibitors may inhibit insulin secretion. Treatment of gastrinoma include both proton pump inhibitors (PPIs) and histamine H2 – receptor antagonists. In patients with glucagonomas hyperglycaemia can be controlled using insulin and oral blood glucose lowering drugs. In malignant glucagonomas smatostatin analogues are effective in controlling necrolytic migratory erythemia. Severe cases of the VIP-oma syndrome require supplementation of fluid losses. Octreotide reduce tumoral VIP secretion and control secretory diarrhoea.

Content of RNA originating from thyroid in washouts from fine-needle and core-needle aspiration biopsy - preliminary study.

INTRODUCTION In the evaluation of molecular markers in washouts from fine-needle aspiration biopsy (FNAB) the extremely small amount of material can be a major problem. Some authors tried to use washouts from core-needle aspiration biopsy (CNABs) to gain more material from larger needles. However, according to some studies, CNAB samples are commonly contaminated with blood. The aim of our study was to evaluate the proportion of nucleic acids from thyroid cells in washouts from FNAB and CNAB by measuring the relative expression of cytokeratin 17 (KRT17) on the mRNA level. MATERIAL AND METHODS Relative expression of KRT17 and GADPH (reference gene) in washouts from FNAB and CNAB was measured using real-time PCR technique and compared to the results from surgical specimens. RESULTS Surgical specimens form 22 nodules, FNAB samples from 20 lesions and CNAB samples from 24 lesions were analysed. The median difference in cycle threshold (Ct) between FNAB samples and surgical specimens was 3.3 (p = 0.047). In CNAB samples KRT17 was undetectable in most cases (median incalculable; proportion of samples with undetectable KRT17 significantly higher than in FNAB samples). CONCLUSIONS Samples obtained with different biopsy techniques had different proportions of contents. The proportionally low content of epithelial cells in CNAB can result in underestimated expression of molecular markers of malignancy. Consequently, the risk of malignancy or unfavourable prognosis can also be underestimated. To conclude, results obtained from samples gained with one biopsy technique cannot be directly related to thresholds, and generally with experiences gained with other techniques, because it can lead to incorrect clinical interpretation of the results. (Endokrynol Pol 2016; 67 (6): 550-553).

Genomic markers of ovarian adenocarcinoma and its relevancy to the effectiveness of chemotherapy

Ovarian cancer is the eighth most common cancer and the seventh highest cause of cancer-associated mortality in women worldwide. It is the second highest cause of mortality among female reproductive malignancies. The current standard first-line treatment for advanced ovarian cancer includes a combination of surgical debulking and standard systemic platinum-based chemotherapy with carboplatin and paclitaxel. Although a deeper understanding of this disease has been attained, relapse occurs in 70% of patients 18 months subsequent to the first-line treatment. Therefore, it is crucial to develop a novel drug that effectively affects ovarian cancer, particularly tumors that are resistant to current chemotherapy. The aim of the present study was to identify genes whose expression may be used to predict survival time or prognosis in ovarian cancer patients treated with chemotherapy. Gene or protein expression is an important issue in chemoresistance and survival prediction in ovarian cancer. In the present study, the research group consisted of patients treated at the Surgical Clinic of the Gynecology and Obstetrics Gynecological Clinical Hospital, Poznan University of Medical Sciences (Poznan, Poland) between May 2006 and November 2014. Additional eligibility criteria were a similar severity (International Federation of Gynecolgy and Obstetrics stage III) at the time of diagnosis, treatment undertaken in accordance with the same schedule, and an extremely good response to treatment or a lack of response to treatment. The performance of the OncoScan® assay was evaluated by running the assay on samples obtained from the four patients and by following the recommended protocol outlined in the OncoScan assay manual. The genomic screening using Affymetrix OncoScan Arrays resulted in the identification of large genomic rearrangements across all cancer tissues. In general, chromosome number changes were detected in all examined tissues. The OncoScan arrays enabled the identification of ~100 common somatic mutations. Chemotherapy response in ovarian cancer is extremely complex and challenging to study. The present study identified specific genetic alterations associated with ovarian cancer, but not with response for treatment.

Genomic mapping of pathways in endometrial adenocarcinoma and a gastrointestinal stromal tumor located in Meckel's diverticulum

The present study reports the case of a 71-year-old female patient diagnosed with endometrial adenocarcinoma, which was confirmed by histopathology. In the course of performing an elective hysterectomy with adnexa removal, a solid tumor located in Meckels diverticulum (MD) was identified and excised. Due to the unique nature of the lesion, the tumor tissue underwent broad mapping of any genomic alterations once the histopathological examination was completed. The genetic testing was conducted using a high-resolution microarray and resulted in the identification of 45 genomic abnormalities, including 4 chromosomal aneuploidies. Within those regions, alterations of 87 known cancer genes were assigned. The involvement of v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog gene alteration was noted to be a key player for triggering gastrointestinal stromal tumor transformation for this unusual case. A total of 12 genes, showing mutual interaction in different cancer types or involved in diverse cellular processes, were identified. These reported data may shed light on the carcinogenesis of a rare MD tumor.

Clinical features of gastroenteropancreatic tumours

Gastroenteropancreatic (GEP) endocrine tumours (carcinoids and pancreatic islet cell tumours) are composed of multipotent neuroendocrine cells that exhibit a unique ability to produce, store, and secrete biologically active substances and cause distinct clinical syndromes. The classification of GEP tumours as functioning or non-functioning is based on the presence of symptoms that accompany these syndromes secondary to the secretion of hormones, neuropeptides and/or neurotransmitters (functioning tumours). Non-functioning tumours are considered to be neoplasms of neuroendocrine differentiation that are not associated with obvious symptoms attributed to the hypersecretion of metabolically active substances. However, a number of these tumours are either capable of producing low levels of such substances, which can be detected by immunohistochemistry but are insufficient to cause symptoms related to a clinical syndrome, or alternatively, they may secrete substances that are either metabolically inactive or inappropriately processed. In some cases, GEP tumours are not associated with the production of any hormone or neurotransmitter. Both functioning and non-functioning tumours can also produce symptoms due to mass effects compressing vital surrounding structures. Gastroenteropancreatic tumours are usually classified further according to the anatomic site of origin: foregut (including respiratory tract, thymus, stomach, duodenum, and pancreas), midgut (including small intestine, appendix, and right colon), and hindgut (including transverse colon, sigmoid, and rectum). Within these subgroups the biological and clinical characteristics of the tumours vary considerably, but this classification is still in use because a significant number of previous studies, mainly observational, have used it extensively.