Identification of novel genes involved in the pathophysiology of adrenocortical tumorigenesis

Adrenocortical tumors (ACT) are relatively common, with a prevalence of 6% in the aging population. They can be divided into benign adenomas (ACA) and malignant carcinomas (ACC). Tumors responding to luteinizing hormone (LH)/choriogonadotropin (CG) belong to a subgroup of hormonally active ACTs. Such tumors may develop during the chronic elevation of LH (menopause) or hCG (pregnancy), and result in cortisol, aldosterone, or/and androgen overproduction. In the present thesis, transgenic mice expressing SV40Tag oncogene under the inhibin α promoter (inhα/Tag), were utilized to identify novel biomarker genes and unravel the molecular mechanisms in the gonadotropin-dependent adrenocortical tumorigenesis. The effects of GnRH antagonist treatment were investigated on human and mouse adrenocortical cell lines in vitro, and on mouse adrenocortical tumors in vivo. Finally, the role of LHCGR in the pregnancy-induced Cushing syndrome was characterized in a case study. Estrogen receptor alpha (ESR1) expression was identified to be associated with the adrenocortical tumors in inhα/Tag mice, whereas several other molecules, such as GRB10, RERG, GNAS, and NFATC2, were linked to healthy adrenal tissue. Gonadotropin-dependent adrenocortical tumors of inhα/Tag mice were found to originate from zona fasciculata and LHCGR to be a prerequisite for their onset, presumably through the cell fate reprogramming by GATA6 to GATA4 transcription factor switch. After induction, GATA4 took over the transcriptional control in the adrenal gland, prompting the tumor progression and gonadal-like phenotype, and sequentially adrenocortical tumors became independent of LHCGR signaling. The GnRH antagonist cetrorelix acetate acted directly on human and mouse adrenocortical tumor cells, inducing apoptosis. Moreover, it appeared that chronically high LH/hCG concentrations promoted the transformation of LHCGR-positive cells into LH/hCG-responsive adrenocortical cells, which gave rise to the cortisol and androgen-producing hyperplastic cells in human, and estrogen-producing neoplasms in mice. In conclusion, LH/LHCGR signaling plays a crucial role in the induction of some adrenocortical tumors. Regulatory role of GATA4 was shown to be required for tumor progression in mice. Cetrorelix acetate could be considered for improving the ACT therapy, either through its systemic or direct action, or their combined effects on adrenocortical tumor cells

Endothelial dysfunction and ADMA in pathogenesis of arterial hypertension

Choroby układu sercowo-naczyniowego stanowią obecnie jeden z najistotniejszych problemów zdrowotnych populacji światowej. Istotnym elementem patologii obserwowanej w tej grupie schorzeń jest dysfunkcja śródbłonka. Jednym z najsilniejszych śródbłonkowych czynników wazodylatacyjnych jest tlenek azotu (NO), wykazujący także właściwości przeciwzapalne i antyagregacyjne. Pośredniczony przez NO, zależny od śródbłonka rozkurcz naczynia odgrywa istotną rolę w regulacji napięcia ściany naczynia. Dysfunkcja śródbłonkowych szlaków metabolicznych zależnych od NO może istotnie zwiększać ryzyko sercowo-naczyniowe, stanowiąc jednocześnie istotne ogniwo patogenezy nadciśnienia tętniczego. Asymetryczna dimetyloarginina (ADMA) jest endogennym inhibitorem kompetycyjnym syntazy NO, który poprzez zmniejszenie biodostępności NO, może prowadzić do dysfunkcji śródbłonka. ADMA jest nie tylko markerem, lecz także istotnym czynnikiem biorącym udział w patogenezie dysfunkcji śródbłonka, a jej podwyższone stężenie stwierdzano w przebiegu wielu chorób układu sercowo-naczyniowego, w tym również w nadciśnieniu tętniczym. Niniejsza praca jest przeglądem bieżących informacji dotyczących dysfunkcji śródbłonka i ADMA w powiązaniu z patogenezą nadciśnienia tętniczego, a także możliwości interwencji farmakologicznych prowadzących do zmiany stężenia ADMA.Cardiovascular diseases (CVD) are today one of the major healthcare problems of the world population. Endothelial dysfunction (ED) reflects prominent abnormalities observed in these disorders. Nitric oxide is one of the most potent endothelial vasodilators and has also anti-inflammatory and antithrombotic properties. NO-mediated endothelium - dependent vasodilatation plays a crucial role in regulation of vascular tone. Impaired activity of NO-dependent metabolic pathways in endothelium may increase the risk of CVD and may be an important pathophysiological link leading to development of arterial hypertension (HTA). Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of NO synthase, which may cause ED due to decreased NO-availability. ADMA may be not only a marker, but also an active mediator in pathogenesis of CVD. Its levels have been shown to be elevated in variety of disorders, e.g. in hypertension. This article gives a brief review of contemporary state of knowledge regarding ED, ADMA and HTA. Pathophysiology of ED and HTA and possible pharmacological interventions aiming the modulation of ADMA level are discussed

Influence of vitamin D on fertility and fetal development – review of literature

Summary Cholecalciferol was qualified by FDA (Food and Drug Administration) into the A category if administered in recommended doses and to the D category if the doses exceed RDA (Recommended Dietary Allowance). There are very divergent opinions among researchers concerning the optimal daily dose of vitamin D-according to some of them, the optimal dose of vitamin D should exceed 400 UI/24h. On the other hand, there is no data to estimate the optimal dose and to formulate recommendations. It is necessary to conduct research on animal models to fully comprehend the symptoms and syndromes caused by excess or deficiency of cholecalciferol. However, the conclusions of the research done on animals should not be over-generalized. Moreover, some data concerning the influence of vitamin D administered during the pregnancy on fetal development are often ambiguous. All these facts are the reason why recommendations of vitamin D supplementation in pregnancy still remain uncertain and need thorough investigation

Modeling glioblastoma heterogeneity as a dynamic network of cell states

Tumor cell heterogeneity is a crucial characteristic of malignant brain tumors and underpins phenomena such as therapy resistance and tumor recurrence. Advances in single-cell analysis have enabled the delineation of distinct cellular states of brain tumor cells, but the time-dependent changes in such states remain poorly understood. Here, we construct quantitative models of the time-dependent transcriptional variation of patient-derived glioblastoma (GBM) cells. We build the models by sampling and profiling barcoded GBM cells and their progeny over the course of 3\ua0weeks and by fitting a mathematical model to estimate changes in GBM cell states and their growth rates. Our model suggests a hierarchical yet plastic organization of GBM, where the rates and patterns of cell state switching are partly patient-specific. Therapeutic interventions produce complex dynamic effects, including inhibition of specific states and altered differentiation. Our method provides a general strategy to uncover time-dependent changes in cancer cells and offers a way to evaluate and predict how therapy affects cell state composition

Resistance to acetylsalicylic acid - pathophysiology and clinical implications

Choroby układu sercowo-naczyniowego są jedną z najczęstszych przyczyn zgonów i długotrwałej niepełnosprawności. Kwas acetylosalicylowy to bardzo skuteczny lek przeciwpłytkowy, zmniejszający ryzyko zawału serca, udaru mózgu i zgonu o około 25% w grupie osób, u których ryzyko sercowo-naczyniowe jest zwiększone. Wyniki badań klinicznych jednoznacznie wskazują na wysoką skuteczność kwasu acetylosalicylowego w profilaktyce pierwotnej i wtórnej incydentów sercowo-naczyniowych, jednakże oporność na ten związek wynosi według różnych szacunków 5-45%. Oporność na kwas acetylosalicylowy jest definiowana na podstawie objawów klinicznych (incydent zakrzepowy podczas jego stosowania), jak również za pomocą wielu wskaźników właściwości agregacyjnych płytek. Małe badania obserwacyjne wskazują na istnienie zwiększonego ryzyka sercowo-naczyniowego w subpopulacji osób, u których występuje oporność na kwas acetylosalicylowy. Dotychczas nie poznano w pełni dokładnego mechanizmu oporności, a poza niepełną inhibicją cyklooksygenazy i uwzględnia się także czas trwania terapii, współpracę ze strony pacjenta, przyjmowanie równocześnie innych niesteroidowych leków przeciwzapalnych. W pracy przedstawiono wszystkie znane mechanizmy prowadzące do oporności na kwas acetylosalicylowy oraz konsekwencje kliniczne tego zjawiska. Zwrócono również uwagę na brak jednoznacznych kryteriów określających tę oporność, a także zaleceń wskazujących, w jakim przypadku i kiedy należy przeprowadzić diagnostykę w tym kierunku.Cardiovascular diseases are the most common cause of morbidity and mortality. Acetylsalicylic acid (ASA) is an effective antiplatelet drug which decreases the risk of myocardial infarction, stroke and sudden death by 25% in groups with higher cardiovascular risk. The high efficacy of ASA in primary and secondary prevention of cardiovascular events might be diminished in cases of ASA resistance, as reported in 5-45% of patients. Acetylsalicylic acid resistance is diagnosed by clinical symptoms (thrombotic event during ASA therapy) or markers of platelet aggregation. Small observational have studies revealed higher cardiovascular risk in populations with ASA resistance. Beyond incomplete inhibition of COX-1, mechanisms of ASA resistance are still not well known. Duration of therapy, lack of compliance and drug interactions are considered as possible factors in the development of ASA resistance. Our review describes all known mechanisms of ASA resistance and their clinical consequences. We have also pointed out the need for the development of universally accepted diagnostic criteria and screening guidelines for ASA resistance

Revisiting the expression and function of follicle-stimulation hormone receptor in human umbilical vein endothelial cells

Expression of follicle-stimulation hormone receptor (FSHR) is confined to gonads and at low levels to some extragonadal tissues like human umbilical vein endothelial cells (HUVEC). FSH-FSHR signaling was shown to promote HUVEC angiogenesis and thereafter suggested to have an influential role in pregnancy. We revisited hereby the expression and functionality of FSHR in HUVECs angiogenesis, and were unable to reproduce the FSHR expression in human umbilical cord, HUVECs or immortalized HUVECs (HUV-ST). Positive controls as granulosa cells and HEK293 cells stably transfected with human FSHR cDNA expressed FSHR signal. In contrast to positive control VEGF, FSH treatment showed no effects on tube formation, nitric oxide production, wound healing or cell proliferation in HUVEC/HUV-ST. Thus, it remains open whether the FSH-FSHR activation has a direct regulatory role in the angiogenesis of HUVECs

Revisiting the expression and function of follicle-stimulation hormone receptor in human umbilical vein endothelial cells

Expression of follicle-stimulation hormone receptor (FSHR) is confined to gonads and at low levels to some extragonadal tissues like human umbilical vein endothelial cells (HUVEC). FSH-FSHR signaling was shown to promote HUVEC angiogenesis and thereafter suggested to have an influential role in pregnancy. We revisited hereby the expression and functionality of FSHR in HUVECs angiogenesis, and were unable to reproduce the FSHR expression in human umbilical cord, HUVECs or immortalized HUVECs (HUV-ST). Positive controls as granulosa cells and HEK293 cells stably transfected with human FSHR cDNA expressed FSHR signal. In contrast to positive control VEGF, FSH treatment showed no effects on tube formation, nitric oxide production, wound healing or cell proliferation in HUVEC/HUV-ST. Thus, it remains open whether the FSH-FSHR activation has a direct regulatory role in the angiogenesis of HUVECs.</p