Analysis of the frequency of EGFR, KRAS and ALK mutations in patients with lung adenocarcinoma in Croatia

BACKGROUND: Many studies have been published on the mutational status of patients with lung adenocarcinomas, and great population-based variability in mutation frequencies has been reported. The main objective of the present study was to analyze the EGFR, KRAS and ALK mutation status in a representative cohort of patients in Croatia with lung adenocarcinomas and to correlate the mutational status with clinical data. ----- METHODS: All patients who were newly diagnosed within 6 months with histologically proven primary lung adenocarcinomas were included. Mutational analyses for EGFR and KRAS mutations were performed in a cobas z 480 analyzer. ALK immunohistochemistry was performed using the D5F3 clone on Benchmark XT instrument. Clinical data were obtained from the medical records. ----- RESULTS: Of the 324 patients, 59.9 % were male. At the time of diagnosis, the patients ranged in age range from 35 to 88 years (median 63 years). Most of the patients were current smokers or former smokers (77.2 %). EGFR mutations were found in 15.7 % of the patients, and of these mutations, exon 19 deletion was the most common (45.1 %). KRAS mutations were present in 34.9 % of the patients, while 4.1 % of patients were ALK-positive. The statistical significance of the presence of mutations was detected for both gender and smoking. ----- CONCLUSION: The detected mutation rates demonstrated a slightly higher prevalence of KRAS mutations, but not a higher prevalence of EGFR mutations or ALK gene rearrangement, in comparison with the rates found in other European countries. EGFR and ALK mutational status showed a statistically significant correlation with gender as well as with smoking, while KRAS mutation status showed a statistically significant correlation only with smoking

Imunoterapija u tretmanu ne-sitnostaničnog raka pluća

Lung cancer is leading cause of death among malignant disease Worldwide and it is responsible for more than 1, 5 million deaths each year. Lung cancer is divided in two major groups: non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). Despite significant improvements, for vast majority of patients chemotherapy still remains the treatment of choice in the first line setting. Progress over the last decade has led to the recognition of immunoevasion as of the leading hallmarks of cancer development. Clinical development was focused on immune checkpoint inhibitors, cytotoxic T-lymphocyte–associated antigen 4 (CTLA4) and programmed death (PD1/PD-L1) pathway. Programmed death 1 protein is another T-cell coinhibitory receptor with a structure similar to that of CTLA-4 but with a distinct biologic function and ligand specificity and it is stimulated with PD-L1. PD-1 or PD-L1 blockade with drugs like nivolumab, pembrolizumab or atezolizumab resulted in superior efficacy comparing to standard chemotherapy in first-line setting. In patient with high PD-L1 expression (50% or more) pemborlizumab should be treatment of choice in first-line setting. PD-L1 expression is at the moment only available biomarker who can predict response to immune checkpoint inhibitors.Karcinom pluća vodeći je uzrok smrti od malignih bolesti te je odgovoran za više od 1,5 milijuna smrti. U većine bolesnika osnovu liječenja karcinoma pluća čini kemoterapija temeljena na platini. Unazad desetak godina, brojna se istraživanje provode o ulozi imunološkog sutava u karcinogenezi. Tumori uspjevaju izbjeći nadzor imunološkog sustava te tako rasti i metastazirati. Koncpet aktivacije imunološkog sutava iznimno je zanimljiv te se pokazalo da blokatori kontrolnih točaka pokazauju dobro učinkovitost u liječenju solidnih tumora poput melanoma ili bubrega. U zadnjih pet godina, inhibitori kontrolnih točaka nivolumab, pembrolizumab i atezoilizumab pokazali su se kao iznimno učinkoviti lijekovi u liječenju bolesnika s karcinomom pluća ranije liječenih klasičnom kemoterapijom. Pembrolizumab se također pokazao kao iznimno učinkovit u liječenju bolesnika sa visokom PD-L1 ekspresijom (PD-L1≥50%) u prvoj liniji liječenja. Ono što je nužno je pronaći biomarker koji bi ukazivao koji će bolesnici regirati na liječenje imunoterapijom. Danas znamo da što je viša ekspresija PD-L1 bolji je odgovor na imunoterapiju, ali i negativni bolesnici mogu odgovoriti na liječenje. Imunoterapija inhibitorima kontrolnih točaka danas je standard u drugoj liniji liječenja karcinoma pluća malih stanica, te je pembrolizumab najbolja opcija za liječenje visoko pozitivnih PD-L1 bolesnika u prvoj liniji liječenja

Mitochondrial unfolded protein response, mitophagy and other mitochondrial quality control mechanisms in heart disease and aged heart

Mitochondria are involved in crucial homeostatic processes in the cell: the production of adenosine triphosphate and reactive oxygen species, and the release of pro-apoptotic molecules. Thus, cell survival depends on the maintenance of proper mitochondrial function by mitochondrial quality control. The most important mitochondrial quality control mechanisms are mitochondrial unfolded protein response, mitophagy, biogenesis, and fusion-fission dynamics. This review deals with mitochondrial quality control in heart diseases, especially myocardial infarction and heart failure. Some previous studies have demonstrated that the activation of mitochondrial quality control mechanisms may be beneficial for the heart, while others have shown that it may lead to heart damage. Our aim was to describe the mechanisms by which mitochondrial quality control contributes to heart protection or damage and to provide evidence that may resolve the seemingly contradictory results from the previous studies

Klinička obilježja i preživljenje bolesnika s malignim mezoteliomom pleure – iskustvo jednog centra

Maligni pleuralni mezoteliom rijetka je i agresivna primarna novotvorina mezotelnih stanica pleure. Glavni rizični čimbenik je izloženost azbestu, najčešće uz latenciju od 30–50 godina. Unatoč terapiji medijan preživljenja je od 4 do 18 mjeseci, ovisno o izvoru. Cilj istraživanja bio je odrediti karakteristike bolesnika oboljelih od malignoga pleuralnog mezotelioma te ishode njihova liječenja na KBC Zagreb u razdoblju od 1999. do 2012. godine. Ispitane su karakteristike 101 bolesnika dijagnosticiranog i liječenog na KBC Zagreb u razdoblju od rujna 1999. do rujna 2012. Analizirali smo ukupno preživljenje (OS), preživljenje bez progresije bolesti (PFS) te preživljenje ovisno o histološkom podtipu tumora, dobi, kliničkom stadiju, pleurodezi talkom i modalitetu liječenja. Koristili smo Kaplan-Meierovu metodu za izradu krivulje preživljenja. 89 bolesnika bilo je muškog, 12 ženskog spola, a medijan dobi 62 godine. Prema TNM klasifikaciji stadij IV je utvrđen kod 69,3%, stadij III kod 26,73%, te stadij II kod samo 3,96% bolesnika. 73,26% bolesnika imalo je epiteloidni, 4,95% sarkomatoidni, 1% mješoviti te 20,79% NOS (nespecifirani) histološki podtip bolesti. Kirurški je liječeno 14,85% bolesnika, kemoterapijom 55,44%, dok je radioterapija primijenjena kod 9,9% bolesnika. Medijan OS iznosi 11 mjeseci, dok je PFS 10 mjeseci. Prema histološkom podtipu medijan OS iznosio je 11 mjeseci za epiteloidni, 12,5 za NOS te 5,5 za sarkomatoidni, a prema kliničkom stadiju 7 mjeseci za stadij II, 17,5 mjeseci za stadij III te 11 mjeseci za stadij IV. Medijan OS bio je dulji u skupini bolesnika mlađoj od 65 godina (12 naspram 8,5 mjeseci, p=0.28), skupini sa učinjenom pleurodezom (13 naspram 7,5 mjeseci, p=0.06) te u skupini liječenoj kemoterapijom (12,5 naspram 7,5 mjeseci, p=0.10). Očekivano preživljenje bolesnika oboljelih od malignoga pleuralnog mezotelioma u Republici Hrvatskoj u skladu je s podatcima iz literature. Ispitane metode liječenja relativno skromno utječu na preživljenje bolesnika

Analysis of the transcription factors SOX2, OCT4 and NANOG expression and the PI3K/AKT/BCL2 pathway in malignant pleural mesothelioma

Uvod: Maligni pleuralni mezoteliom je zloćudni tumor mezotelnog porijekla, čija dijagnostika i liječenje još uvijek predstavljaju izazov. OCT4, NANOG i SOX2 su čimbenici pluripotencije čija je aktivnost ključna za procese dijeljenja i sazrijevanja stanica tijekom embrionalnog razvoja, a koji se istražuju radi njihove uloge tokom onkogeneze. Signalni put PI3K/AKT/BCL2 jedna je od bolje proučenih kaskada prijenosa unutarstaničnih signala, čijom aktivnošću stanice raka izbjegavaju procese apoptoze i time si omogućavaju preživljenje. Materijali i metode: U istraživanju su korišteni uzorci pacijenata s malignim pleuralnim mezoteliomom. Učinjene su imuohistokemijske i PCR analize radi detekcije proteina odnosno gena čimbenika pluripotencije OCT4, NANOG i SOX2 i puta PI3K/AKT/BCL2. Rezultati: U ovom smo istraživanju dokazali aktivnost proteina i gena čimbenika pluripotencije u mezoteliomu pleure te u zdravom mezotelu. Aktivnost gena NANOG, SOX2 i OCT4 bila je viša u stanicama mezotela, dok je izraženost proteina bila viši u mezoteliomskim stanicama. Izraženost NANOG-a povezan je sa slabije diferenciranim podtipom mezotelioma. U istraživanju smo pokazali izraženost PI3K i AKT i BCL2 u mezoteliomu i u pleuri, pri čemu je aktivnost gena bila viša u stanicama pleure, a proteina AKT i BCL2 u stanicama mezotelioma. Najlošije preživljenje imali su pacijenti sa sarkomatoidnim i pleomorfnim mezoteliomom, dok utjecaj izraženosti NANOG, OCT4 i SOX 2 te PI3K i AKT na preživljenje nije dokazan. Zaključak: Zaključujemo kako su navedeni signalni mehanizmi aktivni u mezoteliomu pleure, a potrebna su dodatna istraživanja radi otkrivanja preciznih prediktivnih faktora u ovoj bolesti.Introduction: Malignant pleural mesothelioma is a malignant tumor of mesothelial origin, the diagnosis and treatment of which still represent a challenge. OCT4, NANOG and SOX2 are pluripotency factors whose activity is crucial for the processes of cell division and maturation during embryonic development, and which are being investigated for their role during oncogenesis. The PI3K/AKT/BCL2 signaling pathway is one of the better-studied intracellular signal transmission cascades, with the activity of which cancer cells avoid the processes of apoptosis and thus enable their survival. Materials and methods: The study used samples from patients with malignant pleural mesothelioma. Immunohistochemical and PCR analyzes were performed to detect proteins or genes of the pluripotency factors OCT4, NANOG and SOX2 and the PI3K/AKT/BCL2 pathway. Results: In this study, we demonstrated the activity of pluripotency factor proteins and genes in pleural mesothelioma and in healthy mesothelium. NANOG, SOX2 and OCT4 gene activity was higher in mesothelial cells, while protein expression was higher in mesothelioma cells. NANOG expression is associated with a less differentiated subtype of mesothelioma. In the research, we showed the expression of PI3K and AKT and BCL2 in mesothelioma and in the pleura, whereby gene activity was higher in pleural cells, and AKT and BCL2 proteins in mesothelioma cells. Patients with sarcomatoid and pleomorphic mesothelioma had the worst survival, while the influence of the expression of NANOG, OCT4 and SOX 2 and PI3K and AKT on survival was not proven. Conclusion: We conclude that the above signaling mechanisms are active in pleural mesothelioma, and additional research is needed to reveal precise predictive factors in this disease

Analysis of the transcription factors SOX2, OCT4 and NANOG expression and the PI3K/AKT/BCL2 pathway in malignant pleural mesothelioma

Uvod: Maligni pleuralni mezoteliom je zloćudni tumor mezotelnog porijekla, čija dijagnostika i liječenje još uvijek predstavljaju izazov. OCT4, NANOG i SOX2 su čimbenici pluripotencije čija je aktivnost ključna za procese dijeljenja i sazrijevanja stanica tijekom embrionalnog razvoja, a koji se istražuju radi njihove uloge tokom onkogeneze. Signalni put PI3K/AKT/BCL2 jedna je od bolje proučenih kaskada prijenosa unutarstaničnih signala, čijom aktivnošću stanice raka izbjegavaju procese apoptoze i time si omogućavaju preživljenje. Materijali i metode: U istraživanju su korišteni uzorci pacijenata s malignim pleuralnim mezoteliomom. Učinjene su imuohistokemijske i PCR analize radi detekcije proteina odnosno gena čimbenika pluripotencije OCT4, NANOG i SOX2 i puta PI3K/AKT/BCL2. Rezultati: U ovom smo istraživanju dokazali aktivnost proteina i gena čimbenika pluripotencije u mezoteliomu pleure te u zdravom mezotelu. Aktivnost gena NANOG, SOX2 i OCT4 bila je viša u stanicama mezotela, dok je izraženost proteina bila viši u mezoteliomskim stanicama. Izraženost NANOG-a povezan je sa slabije diferenciranim podtipom mezotelioma. U istraživanju smo pokazali izraženost PI3K i AKT i BCL2 u mezoteliomu i u pleuri, pri čemu je aktivnost gena bila viša u stanicama pleure, a proteina AKT i BCL2 u stanicama mezotelioma. Najlošije preživljenje imali su pacijenti sa sarkomatoidnim i pleomorfnim mezoteliomom, dok utjecaj izraženosti NANOG, OCT4 i SOX 2 te PI3K i AKT na preživljenje nije dokazan. Zaključak: Zaključujemo kako su navedeni signalni mehanizmi aktivni u mezoteliomu pleure, a potrebna su dodatna istraživanja radi otkrivanja preciznih prediktivnih faktora u ovoj bolesti.Introduction: Malignant pleural mesothelioma is a malignant tumor of mesothelial origin, the diagnosis and treatment of which still represent a challenge. OCT4, NANOG and SOX2 are pluripotency factors whose activity is crucial for the processes of cell division and maturation during embryonic development, and which are being investigated for their role during oncogenesis. The PI3K/AKT/BCL2 signaling pathway is one of the better-studied intracellular signal transmission cascades, with the activity of which cancer cells avoid the processes of apoptosis and thus enable their survival. Materials and methods: The study used samples from patients with malignant pleural mesothelioma. Immunohistochemical and PCR analyzes were performed to detect proteins or genes of the pluripotency factors OCT4, NANOG and SOX2 and the PI3K/AKT/BCL2 pathway. Results: In this study, we demonstrated the activity of pluripotency factor proteins and genes in pleural mesothelioma and in healthy mesothelium. NANOG, SOX2 and OCT4 gene activity was higher in mesothelial cells, while protein expression was higher in mesothelioma cells. NANOG expression is associated with a less differentiated subtype of mesothelioma. In the research, we showed the expression of PI3K and AKT and BCL2 in mesothelioma and in the pleura, whereby gene activity was higher in pleural cells, and AKT and BCL2 proteins in mesothelioma cells. Patients with sarcomatoid and pleomorphic mesothelioma had the worst survival, while the influence of the expression of NANOG, OCT4 and SOX 2 and PI3K and AKT on survival was not proven. Conclusion: We conclude that the above signaling mechanisms are active in pleural mesothelioma, and additional research is needed to reveal precise predictive factors in this disease

Analysis of the transcription factors SOX2, OCT4 and NANOG expression and the PI3K/AKT/BCL2 pathway in malignant pleural mesothelioma

Uvod: Maligni pleuralni mezoteliom je zloćudni tumor mezotelnog porijekla, čija dijagnostika i liječenje još uvijek predstavljaju izazov. OCT4, NANOG i SOX2 su čimbenici pluripotencije čija je aktivnost ključna za procese dijeljenja i sazrijevanja stanica tijekom embrionalnog razvoja, a koji se istražuju radi njihove uloge tokom onkogeneze. Signalni put PI3K/AKT/BCL2 jedna je od bolje proučenih kaskada prijenosa unutarstaničnih signala, čijom aktivnošću stanice raka izbjegavaju procese apoptoze i time si omogućavaju preživljenje. Materijali i metode: U istraživanju su korišteni uzorci pacijenata s malignim pleuralnim mezoteliomom. Učinjene su imuohistokemijske i PCR analize radi detekcije proteina odnosno gena čimbenika pluripotencije OCT4, NANOG i SOX2 i puta PI3K/AKT/BCL2. Rezultati: U ovom smo istraživanju dokazali aktivnost proteina i gena čimbenika pluripotencije u mezoteliomu pleure te u zdravom mezotelu. Aktivnost gena NANOG, SOX2 i OCT4 bila je viša u stanicama mezotela, dok je izraženost proteina bila viši u mezoteliomskim stanicama. Izraženost NANOG-a povezan je sa slabije diferenciranim podtipom mezotelioma. U istraživanju smo pokazali izraženost PI3K i AKT i BCL2 u mezoteliomu i u pleuri, pri čemu je aktivnost gena bila viša u stanicama pleure, a proteina AKT i BCL2 u stanicama mezotelioma. Najlošije preživljenje imali su pacijenti sa sarkomatoidnim i pleomorfnim mezoteliomom, dok utjecaj izraženosti NANOG, OCT4 i SOX 2 te PI3K i AKT na preživljenje nije dokazan. Zaključak: Zaključujemo kako su navedeni signalni mehanizmi aktivni u mezoteliomu pleure, a potrebna su dodatna istraživanja radi otkrivanja preciznih prediktivnih faktora u ovoj bolesti.Introduction: Malignant pleural mesothelioma is a malignant tumor of mesothelial origin, the diagnosis and treatment of which still represent a challenge. OCT4, NANOG and SOX2 are pluripotency factors whose activity is crucial for the processes of cell division and maturation during embryonic development, and which are being investigated for their role during oncogenesis. The PI3K/AKT/BCL2 signaling pathway is one of the better-studied intracellular signal transmission cascades, with the activity of which cancer cells avoid the processes of apoptosis and thus enable their survival. Materials and methods: The study used samples from patients with malignant pleural mesothelioma. Immunohistochemical and PCR analyzes were performed to detect proteins or genes of the pluripotency factors OCT4, NANOG and SOX2 and the PI3K/AKT/BCL2 pathway. Results: In this study, we demonstrated the activity of pluripotency factor proteins and genes in pleural mesothelioma and in healthy mesothelium. NANOG, SOX2 and OCT4 gene activity was higher in mesothelial cells, while protein expression was higher in mesothelioma cells. NANOG expression is associated with a less differentiated subtype of mesothelioma. In the research, we showed the expression of PI3K and AKT and BCL2 in mesothelioma and in the pleura, whereby gene activity was higher in pleural cells, and AKT and BCL2 proteins in mesothelioma cells. Patients with sarcomatoid and pleomorphic mesothelioma had the worst survival, while the influence of the expression of NANOG, OCT4 and SOX 2 and PI3K and AKT on survival was not proven. Conclusion: We conclude that the above signaling mechanisms are active in pleural mesothelioma, and additional research is needed to reveal precise predictive factors in this disease

Mitochondrial ROS Induce Partial Dedifferentiation of Human Mesothelioma via Upregulation of NANOG

The expression of pluripotency factors is a key regulator of tumor differentiation status and cancer stem cells. The purpose of this study was to examine the expression of pluripotency factors and differentiation status of human mesothelioma and the role of mitochondria in their regulation. We tested the expression of OCT4/POU5F1, NANOG, SOX2, PI3K-AKT pathway and BCL2 genes and proteins in 65 samples of human mesothelioma and 19 samples of normal mesothelium. Mitochondrial membrane potential, reactive oxygen species (ROS) generation and expression of pluripotency factors were also tested in human mesothelioma cell line. Human mesothelium and mesothelioma expressed SOX2, NANOG, PI3K and AKT genes and proteins and POU5F1 gene, whereby NANOG, SOX2 and phosphorylated (activated) AKT were upregulated in mesothelioma. NANOG protein expression was elevated in less differentiated samples of human mesothelioma. The expression of genes of PI3K-AKT pathway correlated with pluripotency factor genes. Mesothelioma cells had functional, but depolarized mitochondria with large capacity to generate ROS. Mitochondrial ROS upregulated NANOG and mitoTEMPO abrogated it. In conclusion, human mesothelioma displays enhanced expression of NANOG, SOX2 and phosphorylated AKT proteins, while elevated NANOG expression correlates with poor differentiation of human mesothelioma. Mitochondria of mesothelioma cells have a large capacity to form ROS and thereby upregulate NANOG, leading to dedifferentiation of mesothelioma