Piotr Glinicki

The diagnosis and classification of adrenal tumors remain a major challenge that involves various fields of medicine [1]. New opportunities in biochemical and molecular testing involve various omics-based techniques that enable the comprehensive analysis of numerous analytes, compounds, proteins, peptides, and nucleic acids in various biological matrices; this comprehensive analysis allows for an assessment of the utility of these tests throughout the entire diagnostic and therapeutic process [2][3][4][5]. Moreover, new imaging techniques allow for a more detailed assessment of pathological changes observed in the diagnosis of adrenal tumors.The current Research Topic remains of high scientific and clinical interest and includes 12 articles.Bernardi et al. [6] described a novel and previously not reported germline variant in the SDHB gene in a patient with a metastatic sympathetic paraganglioma (PGL) presented with hypertension, tachycardia and hyperhidros. Debulking surgery as well as genetic testing were scheduled. The authors concluded that the clinical features and immunohistochemical findings regarding the SDHB gene provided a solid basis for classifying the new germline SDHB c.314T>A genetic variant as a class 4 variant, i.e., "likely pathogenic".Pauzi et al. [7] presented an article titled "Histopathological spectrum of common aldosterone-driver gene mutant aldosterone-producing adenomas (APAs)," where the authors analyzed 33 APAs with confirmed aldosterone-driver variants (17 KCNJ5 variant APAs, 8 ATP1A1 variant APAs, 6 CACNA1D variant APAs, and 2 CTNNB1 variant APAs) were immunohistochemically stained using H&E, CYP17A1, CYP11B2, KCNJ5, Ki67, β-catenin, and LHCGR antibodies. Results of this study support the suggestion that CYP11B2-guided sequencing of all the currently known aldosteronedriver genes can fine tune current genotype-phenotype histopathology profiles.The review article by Szatko A et al. [8] described the current challenges of biochemical diagnostics (immunoassays) of primary aldosteronism, including screening, case confirmation and subtyping and novel "omics" techniques in the diagnostics (steroidomics and proteomics).Adrenocortical carcnoma (ACC) presenting as pheochromocytoma is rare in the literature. In a case report and mini-review [9] authors reported on ACC cases clinically manifesting as pheochromocytomas, including a novel clinical case of a 59-year-old female.Peng et a. [10], presented the diagnostic value of 18 F-AlF-NOTA-Pentixafor PET/CT in subtyping primary aldosteronism in 88 patients. The CXCR4-targeted 18 F-AlF-NOTA-pentixafor PET/CT was a valuable noninvasive tool for diagnosing unilateral primary aldosteronism, demonstrating high sensitivity and specificity.Mytareli et al. [11] presented an original research to validate the diagnostic performance of selected circulating microRNAs (miR-483-5p, miR-210, miR-335 and miR-22-3p), identified through microRNA profiling studies, as markers of malignancy or cortisol hypersecretion in a cohort of patients with adrenal tumors. This study supports the potential of a liquid biopsy approach as an innovative method for diagnosing and monitoring patients with adrenal tumors, offering a complementary tool to existing diagnostic methods.In a case report and literature review by Woźniak et al. [12] entitled "A 17-hydroxyprogesteronesecreting adrenal adenoma mimicking nonclassical 21-hydroxylase deficiency" highlighted the occurrence of rare causes of benign adrenal adenomas with steroidogenesis disorders, which may lead to a misdiagnosis of congenital adrenal hyperplasia (CAH).Weng et al. [13] explored the value of serum steroid profiling as a complementary biomarker for malignancy diagnosis of ACC other than diameter, and explored the influence of sex and functional status. In this retrospective study, a matched cohort of patients diagnosed with either ACC or adrenocortical adenoms (ACA) based on histopathology was meticulously paired in a 1:1 ratio according to sex, age and functional status. Serum steroid profiling served as a helpful discriminative marker for ACC and ACA, with 11-deoxycortisol being the most valuable one. For other steroid hormones, consideration of sex differences and functional status was crucial. Apsan et al. [14] investigated the relationship between adiponectin, leptin and visfatin concentrations to metabolic risk factors and androgen concentrations in children and young adults with CAH. The conclusion was an inverse relationship between adiponectin and androstenedione which suggests that better CAH control can reduce the risk of insulin resistance and metabolic syndrome. However, a high glucocorticoid dose appeared to increase the risk of insulin resistance, underscoring the delicate balance required when treating CAH. Zheng et al. [15] described a novel PRKAR1A gene variant in Chinese patient with Carney complex and reviewed the literature to enhance the understanding of Carney complex.A mini-review by Olejarz et al. [16] discussed publications from the past two years on changes in lipoprotein concentrations in endocrine disorders and provided an overview of the latest findings in this rapidly evolving and extensive field.

Primary aldosteronism is the most common cause of secondary hypertension, yet most cases remain unrecognized and left without optimal treatment. The diagnostic inertia may be attributed to the lack of specific symptoms, insufficient awareness among physicians, still conflicting indications for screening for primary aldosteronism and first and foremost challenging diagnostics. This review describes the current challenges of biochemical diagnostics of primary aldosteronism, including screening, case confirmation and subtyping. It also discusses immunoassays widely used in assessment of suspected autonomous aldosterone secretion – recent advances in the field and limitations of the method in comparison to the gold standard - liquid chromatography –tandem mass spectrometry. The review focuses on the application of novel “omics” strategies in the diagnostics of primary aldosteronism. Steroidomics and proteomics offer a possibility to simultaneously assess steroids and protein/peptides on a large scale. This multianalyte approach in comparison to the selective quantification of a chosen compound has been proved useful in the diagnostics of primary aldosteronism. It also offers a unique insight into the individual characteristics, underlying mechanisms and even reflects the genetic alterations of primary aldosteronism cases. The “omics” techniques are associated with large amounts of generated data, the interpretation of which may be troublesome and often necessitates the use of artificial intelligence. The novel advances in the biochemical diagnostics of primary aldosteronism, including “omics” techniques, presented in this review may help to address the most emerging problems, increase the number of diagnosed patients and facilitate the choice of an optimal treatment.

Biochemical research in metabolomics and proteomics, supported by bioinformatics techniques based on artificial intelligence (AI).New biochemical methods for the diagnosis and monitoring of patients with adrenal tumors are currently being developed in two parallel directions. The first direction of research involves classical immunological (immunochemical) methods based on the antigen-antibody reaction. In particular, automated immunochemical methods based on the chemiluminescence reaction (CLIA, ECLIA) are being evaluated, primarily as widely available, inexpensive, and easy-to-use screening tools [1]. These studies include the use of these methods as screening tests in the diagnosis of patients with suspected primary aldosteronism (PA), the feasibility and clinical value of assaying various hormones, e.g. aldosterone, direct renin concentration (DRC), plasma renin activity (PRA), cortisol in tests aimed at differentiating between different subtyping of PA, and, thirdly, the feasibility of assaying free hormone fractions in various biological materials (e.g., free cortisol in saliva) [2][3][4]. The other direction involves chromatographic methods. New possibilities for biochemical diagnostics are enabled by metabolomics research using modern analytical techniques: Liquid chromatography-mass spectrometry (LC-MS/MS), Gas chromatography/mass spectrometry (GC/MS) and others. The development of various chromatographic techniques, especially the LC-MS/MS technique, makes it possible to determine multiple hormones during a single analysis (e.g., steroid hormone profile in plasma or daily urine collection, free metanephrines profile in plasma, etc.) in routine diagnostics [5,6]. The introduction of the ability to determine hormone panels using various chromatographic techniques (mainly LC-MS/MS and GC/MS) into biochemical diagnostics enables new studies both in screening diagnosis of adrenal tumor lesions and in other diagnostic and therapeutic areas. An additional unique feature of hormonal profile studies in biological material from patients with adrenal tumors can be their personalized nature, which can be used in the individual diagnostic-therapeutic process in a given group of patients [7,8].The advances in the field of molecular studies in adrenal tumors have been made in the molecular characterization of the pheochromocytoma, adrenal Cushing syndrome, PA, and adrenocortical carcinoma (ACC). As for pheochromocytomas, although some older studies reported a prevalence of germline genetic variants in 10% -15% of the patients, more recent studies performed with NGS technology (Next-generation sequencing technology) describe genetic variants in up to 35% -40% of the patients [9][10][11]. The prevalence of genetic variants in these patients has been increasing over the years since new genes such as CSDE1, H3F3A, MET, MERTK and IRP1, have been discovered [10].Furthermore, recent advances in the studies on pheochromocytomas have revealed new molecular events in these tumors. Events that are linked to their genetic landscape and, therefore, provide new therapeutic platforms. In relation to adrenal Cushing syndrome, one of the most recent discoveries has taken place in patients with primary bilateral macronodular adrenal hyperplasia (PBMAH) with the discovery of the role of variants in KDM1A in GIP-mediated cases of Cushing syndrome. It has been reported that 100% of the patients with PBMAH and GIP-responsive Cushing syndrome had a germline variant in KDM1A, compared with 0% of the patients of the control group [11]. The discovery of genetic alterations such as ARMC5 and KDM1A in PBMAH allows early detection of PBMAH in patients' relatives [12]. Regarding PA, several genetic defects in the germline or somatic state have been identified. Although only 5% of the PA are familial [13], currently it is known that about 90% of aldosterone producing adenomas (APAs) are due to somatic variants in genes encoding ion channels or transporters including KCNJ5, CACNA1D, ATP1A1, and ATP2B3 [14]. In the last years, new somatic variants have been identified, including a new one in CACNA1H [15]. In addition, more recently, co-existence of CTNNB1 with G Protein Subunit Alpha Q (GNAQ)/G Protein Subunit Alpha 11 (GNA11) variants has been documented in 59% of APAs [16]. Finally, advances in genetics of ACC have also been reported. Although there are not many current therapy options directly targeting reported ACC alterations, some studies have detected variants in TP53, BRD9, TERT, CTNNB1, CDK4, FLT4 and MDM2 as potentially targetable genetic alterations in patients with ACC [17]. Utilizing blood-based NGS to characterize genomic alterations in advanced ACC is feasible in over 80% of patients the patients, being 50% of them potentially targetable [18]. Thus, in conclusion, advances in the knowledge of the genetic context of functioning adrenal tumors have allowed a better characterization of these tumors, with important implications in the management of these patients, including the personalization of follow-up and treatment, and its importance in the face of genetic counseling for patients and their relatives.The aim of the first article [19] was to compare the clinical presentation and laboratory hormonal diagnostics in patients with two forms of PA: familial hyperaldosteronism (FH) and primary hyperaldosteronism (PA). The study was a meta-analysis conducted on a systematic review of the literature to identify patients with FH. A total of 360 FH cases (246 FH type I, 73 type II, 29 type III and 12 type IV) and 830 sporadic PA patients (from the SPAIN-ALDO registry) were included in the study. Analysis of the results showed a different clinical presentation of patients with FH-I and III compared to sporadic forms of PA. In this regard, FH-I is characterized by a low prevalence of hypokalemia, while FH-III is characterized by severe aldosterone over-secretion causing hypokalemia in more than 85% of patients. The...

Background/Objectives: Thyroid autoimmunity (TAI) affects about 15% of women of reproductive age and can negatively affect pregnancy outcomes. One possible mechanism for pregnancy complications can be attributed to a disturbed process of placentation caused by thyroid antibodies. To test this hypothesis, placental hormones and angiogenic factors in pregnant women with TAI were evaluated. Methods: Fifty-eight hypothyroid women positive for TPOAb/TgAb, thirty-three hypothyroid women negative for TPOAb/TgAb, and thirty-nine healthy controls were enrolled in this study. Maternal thyroid function tests were established every month throughout pregnancy, and angiogenic placental factors, pro-angiogenic placental growth factor (PlGF); two anti-angiogenic factors, soluble vascular endothelial growth factor receptor 1 (sFlt-1) and soluble endoglin (sEng); and placental hormones, estradiol, progesterone, and hCG, were determined during each trimester. Results: Obstetrical and neonatal outcomes did not differ between the groups. However, several detrimental effects of thyroid antibodies were observed. These included a positive correlation between TgAb and the sEng/PlGF ratio in the first trimester and positive correlations between TPOAb and sFlt-1 and between TgAb and the sFlt-1/PlGF ratio in the third trimester. TgAbs in the first trimester was a risk factor for gestational hypertension and preeclampsia. Conclusions: Our study indicates that TPOAbs and TgAbs can exert a direct harmful effect on placentation, leading to disturbances in the production of placental angiogenic factors and, consequently, to an increased risk of gestational hypertension and preeclampsia.

Recent years brought a remarkable progress on understanding the pathophysiology of rare, neuroendocrine tumours– pheochormocytomas and paragangliomas (PPGLs). Approximately 70% PPGLs are associated with germline or somatic mutation in one of more than 25 identified susceptibility genes, which were grouped into three clusters.Cluster 1 includes genes encoding proteins involved in Krebs cycle and hypoxia pathway – cluster 1 related PPGL are usually located extra-adrenally (often manifest as head and neck paragangliomas), they are also characterized by noradrenergic/dopaminergic biochemical phenotype and increased metastatic risk. Cluster 2 encompasses genes involved in kinase signaling pathway, PPGL related to cluster 2 are predominantly pheochromocytomas with adrenergic biochemical phenotype, the patients rarely develop metastases. The least known cluster 3, associated with deregulation of Wnt signaling pathway, includes exclusively somatic mutations leading to aggressive pheochromocytomas.This review summarizes molecular background and the differences between PPGL clusters regarding clinical course, biochemical phenotype, optimal imaging modalities and targeted molecular therapies for metastatic PPGL. Ostatnie lata przyniosły znaczny postęp w poznaniu patofizjologii rzadkich guzów neuroendokrynnych – guzów chromochłonnych i przyzwojaków (pheochromocytomas and paragangliomas (PPGLs)). Około 70 % PPGL związanych jest z mutacjami: germinalną lub somatyczną w jednym z ponad 25 poznanych genów związanych z rozwojem PPGL. W oparciu o rolę kodowanych białek, geny te zostały podzielone na trzy klastry.Klaster 1 obejmuje geny kodujące białka o aktywności enzymatycznej biorące udział w cyklu Krebsa oraz białka związane ze szlakiem hipoksji. PPGLs związane z mutacjami genów należących do klastra 1 zazwyczaj rozwijają się z autonomicznych zwojów współczulnych lub przywspółczulnych (przyzwojaki głowy i szyi), charakteryzują się zwiększoną sekrecją metabolitów noradrenaliny i/lub dopaminy oraz znacznym ryzykiem tworzenia przerzutów. Klaster 2 to geny kodujące białka związane ze szlakiem sygnałowym kinaz – u chorych z PPGL uwarunkowanymi mutacjami należącymi do tego klastra zazwyczaj stwierdza się guzy chromochłonne, podwyższone stężenie metabolitów adrenaliny w diagnostyce biochemicznej, a rozsiana postać występuje rzadko. Najmniej poznany klaster 3 skupia geny związane ze szlakiem sygnałowym Wnt – wśród tych pacjentów dominują guzy zlokalizowane w nadnerczach o agresywnym przebiegu.W tym artykule poglądowym zostały przedstawione uwarunkowania molekularne podziału PPGLs na klastery, a także kliniczne, biochemiczne, obrazowe oraz terapeutyczne implikacje proponowanej klasyfikacji.

Prolonged exposure to an excess of glucocorticosteroids (GCs), both endogenous and exogenous, leads to a wide range of comorbidities, including cardiovascular, metabolic, psychiatric, and musculoskeletal disorders. The latter comprise osteopenia and osteoporosis leading to skeletal fractures and myopathy. Although endogenous hypercortisolemia is a rare disorder, GCs are among the most frequently prescribed drugs, often administered chronically and despite multiple side effects, impossible to taper off due to therapeutic reasons. The pathophysiology of the effect of GC excess on bone often leads to fractures despite normal or low-normal bone mineral density and it includes direct (mainly disturbance in bone formation processes, through inactivation of the Wnt/beta-catenin signalling pathway) and indirect mechanisms (through suppressing the gonadal and somatotrophic axis, and also through antagonizing vitamin D actions). Glucocorticosteroid-induced fast-twitch, glycolytic muscles atrophy occurs due to increased protein catabolism and impaired synthesis. Protein degradation is a result of activation of the ubiquitin proteasome and the lysosomes stimulated through overexpression of several atrogenes (such as FOXO-1 and atrogin-1). This review will discuss pathophysiology, clinical presentation, prevention, and management of GCinduced osteoporosis (including calcium and vitamin D supplementation, and bisphosphonates) and myopathy associated with GC excess.

Primary aldosteronism (PA) is the most common, but broadly underdiagnosed, form of hormonal hypertension. To improve screening procedures, current biochemical approaches aim to determine newly appreciated angiotensin II (Ang II) and calculate the aldosterone-to-angiotensin II ratio (AA2R). Thus, the aim of this study was to assess the diagnostic performance of these screening tests in comparison to the aldosterone-to-direct renin ratio (ADRR), which is routinely used. Cheap and available ELISA was used for Ang II measurement. To our knowledge, this is the first study of this laboratory method’s usage in PA. The study cohort included 20 PA patients and 80 controls. Ang II concentrations were comparable between PA and non-PA patients (773.5 vs. 873.2 pg/mL, p = 0.23, respectively). The AA2R was statistically significantly higher in PA group when compared with non-PA (0.024 vs. 0.012 ng/dL/pg/mL, p < 0.001). However, the diagnostic performance of the AA2R was significantly worse than that of the ADRR (AUROC 0.754 vs. 0.939, p < 0.01). The sensitivity and specificity of the AA2R were 70% and 76.2%, respectively. Thus, the AA2R was not effective as a screening tool for PA. Our data provide important arguments in the discussion on the unsatisfactory accuracy of renin–angiotensin system evaluation by recently repeatedly used ELISA tests.

Introduction: Pheochromocytoma is a hormonally active tumour originating from neuroendocrine cells of the adrenal medulla. Chromogranin A (CgA) and peptide proSAAS belong to the family of granins and are present in neuroendocrine cells of adrenal medulla, from where they are released to circulation, along with catecholamines. The aim of this study was to assess the usability of proSAAS peptide assay in patients with adrenal pheochromocytoma. Material and methods: 23 patients (13 females and 10 males) with adrenal pheochromocytoma (benign in 18 patients and malignant in 5) confirmed by histopathology examination, and 35 blood donors as a control group. Plasma free metanephrines, CgA, and proSAAS peptide levels were measured in all participants. Results: CgA and proSAAS levels in the group of pheochromocytoma patients vs. the control were: 209 ng/mL and 0.8 ng/mL vs. 59 ng/mL and 0.3 ng/mL (p < 0.001), respectively. The following sensitivity and specificity indexes were obtained from ROC curves for CgA: 83% and 92%, respectively, and for the proSAAS peptide: 39% and 88%, respectively. The combination of 2 parameters: normetanephrine and proSAAS (96% and 100%) had a high diagnostic value, and the value of all determined parameters together (metanephrine, normetanephrine, CgA, and proSAAS) was 100%. Conclusion: A single determination of the proSAAS peptide level is associated with a rather low diagnostic value. But collective determination of CgA and proSAAS may be an additional, valuable tool in biochemical diagnostics of pheochromocytoma. Introduction: Pheochromocytoma is a hormonally active tumour originating from neuroendocrine cells of the adrenal medulla. Chromogranin A (CgA) and peptide proSAAS belong to the family of granins and are present in neuroendocrine cells of adrenal medulla, from where they are released to circulation, along with catecholamines. The aim of this study was to assess the usability of proSAAS peptide assay in patients with adrenal pheochromocytoma. Material and methods: 23 patients (13 females and 10 males) with adrenal pheochromocytoma (benign in 18 patients and malignant in 5) confirmed by histopathology examination, and 35 blood donors as a control group. Plasma free metanephrines, CgA, and proSAAS peptide levels were measured in all participants. Results: CgA and proSAAS levels in the group of pheochromocytoma patients vs. the control were: 209 ng/mL and 0.8 ng/mL vs. 59 ng/mL and 0.3 ng/mL (p < 0.001), respectively. The following sensitivity and specificity indexes were obtained from ROC curves for CgA: 83% and 92%, respectively, and for the proSAAS peptide: 39% and 88%, respectively. The combination of 2 parameters: normetanephrine and proSAAS (96% and 100%) had a high diagnostic value, and the value of all determined parameters together (metanephrine, normetanephrine, CgA, and proSAAS) was 100%. Conclusion: A single determination of the proSAAS peptide level is associated with a rather low diagnostic value. But collective determination of CgA and proSAAS may be an additional, valuable tool in biochemical diagnostics of pheochromocytoma.

Levothyroxine (LT4) is a standard therapy in hypothyroidism; however, its bioavailability and therapeutic effects might be affected by many factors. Data shows that therapy with liquid LT4 characterized by quicker pharmacokinetics provides better thyroid hormones control than tablet LT4. We addressed the quality of life (QoL) and efficacy of the new ethanol-free formula of liquid LT4 (Tirosint^®SOL) treatment in 76 euthyroid patients with primary (PH, n = 46) and central hypothyroidism (CH, n = 30), and compared the results to retrospective data on equivalent doses of tablet L-T4 therapy. After 8 weeks of liquid LT4 therapy, we found a significant improvement in QoL in both PH and CH patients. TSH levels were unaltered in PH patients. Free hormone levels (fT4 and fT3) increased in all the patients, with the exception of fT3 in the CH group. SHBG and low-density lipoprotein (LDL) also improved. Liquid LT4 therapy provided a better thyroid hormone profile and improvement in patients’ QoL than the tablet form, which was possibly due to the more favorable pharmacokinetics profile resulting in better absorption, as suggested by the increased free thyroid hormone levels. In summary, this is the first study addressing the QoL in hypothyroid patients, including primary and central hypothyroidism, treated with liquid LT4 formula in everyday practice.