The role of the metabolite cargo of extracellular vesicles in tumor progression

Metabolomic reprogramming in tumor and stroma cells is a hallmark of cancer but understanding its effects on the metabolite composition and function of tumor-derived extracellular vesicles (EVs) is still in its infancy. EVs are membrane-bound sacs with a complex molecular composition secreted by all living cells. They are key mediators of intercellular communication both in normal and pathological conditions and play a crucial role in tumor development. Although lipids are major components of EVs, most of the EV cargo studies have targeted proteins and nucleic acids. The potential of the EV metabolome as a source for biomarker discovery has gained recognition recently, but knowledge on the biological activity of tumor EV metabolites still remains limited. Therefore, we aimed (i) to compile the list of metabolites identified in tumor EVs isolated from either clinical specimens or in vitro samples and (ii) describe their role in tumor progression through literature search and pathway analysis

A szegedi Fogászati és Szájsebészeti Klinikán diagnosztizált, az orofacialis régiót érintő jóindulatú daganatok és daganatszerű laesiók klinikopatológiai retrospektív epidemiológiai analízise (1960–2014) = A clinicopathological retrospective epidemiological analysis of benign tumors and tumor-like lesions in the oral and maxillofacial region, diagnosed at the University of Szeged, Department of Oral Medicine (1960–2014)

Absztrakt: Bevezetés: Az 54 éves (1960–2014) retrospektív klinikopatológiai analízis a szegedi Fogászati és Szájsebészeti Klinika Orális Medicina Részlegén diagnosztizált jóindulatú orofacialis daganatok és daganatszerű elváltozások utánkövetéses epidemiológiai eredményeinek bemutatására irányult. Anyag és módszer: Összesen 14 661 biopszia történt, amelyek közül 7491 (51,09%) jóindulatú daganatos beteget vontunk be számítógépes vizsgálatunkba. Eredmények: A betegek átlagéletkora 55,3 év, a férfiak száma 2823 (37,7%), a nőké 4668 (62,3%) volt. A férfi : nő arány 1 : 1,65 volt. Az 51–60 éves korcsoportból emelhető ki a legtöbb, 1280 eset (17,1%), és ezen belül 1014 eset (13,6%) alakult ki gyermekkorban és 6477 eset (86,3%) felnőttkorban. Dominálóan több volt a nem neoplasma (6420, 85,7%), mint a neoplasma (1071, 14,3%), ezenfelül pedig több volt a mesenchymalis (5574, 74,4%), mint a nem mesenchymalis (982, 13,1%) daganat. A leggyakoribb daganattípus az irritációs fibroma volt (1806, 32,4%). A gyulladásos/fertőzéses csoportban a legtöbbször granuloma pyogenicumot (465, 8,3%) észleltünk. A cysták közül a mucokele (805, 10,7%), a fejlődési rendellenességek közül pedig a haemangioma (815, 14,6%) fordult elő a leggyakrabban. A daganatok lokalizációját tekintve a legtöbb esetet az ajkon (2081, 27,8%), a gingiván (2024, 27,0%), a buccán (1069, 14,3%), a nyelven (981, 13,1%) és az arcbőrön (695, 9,3%) regisztráltuk. Az orofacialis jóindulatú lágyrész-daganatok döntő többségét a biopsziavételt követően cryo- vagy lézer-, esetenként kombinált (cryo + lézer) kezelésben részesítettük. Következtetés: A jelen vizsgálatban a leggyakoribb jóindulatú tumor az irritációs fibroma volt, és a legtöbb elváltozás az alsó ajkakon fordult elő. A tanulmányok összehasonlító vizsgálatát megnehezítették a diagnosztikai klasszifikációban és a metodológiában alkalmazott különbözőségek, valamint a változó földrajzi és populációs eltérések. Orv Hetil. 2018; 159(37): 1516–1524. | Abstract: Introduction: In a clinicopathological retrospective epidemiological study we investigated benign tumors and tumor-like lesions located in the orofacial region, diagnosed at the Universiy of Szeged, Department of Oral Medicine. Method: During a 54-year period (1960–2014), 14 661 biopsies were taken. The included subjects were 7491 patients diagnosed with benign tumors and tumor-like lesions. Results: The average age of patients was 55.3 years, 2823 (37.7%) patients were male and 4668 (62.3%) female. The male : female ratio was 1 : 1.65. Most of the patients included in the study were aged 51–60 (1280, 17.1%). The number of children was 1014 (13.6%) and the number of adults was 6477 (86.3%). The number of non-neoplasms was 6420 (85.7%), being significantly higher than the number of neoplasms (1071, 14.3%). Most of the lesions were of mesenchymal origin (5574, 77.4%); the number of lesions of non-mesenchymal origin was 982 (13.1%). The most prevalent type of lesions was traumatic fibroma (fibrosis): 1806 (32.4%). The most common lesion type in the group of lesions of infectious/inflammational origin was pyogenic granuloma, the number of which was 465 (8.3%). The most common cystic lesion was mucocele (805, 10.7%). Hemangioma was the most frequent lesion type among developmental anomalies with the number of 815 (14.6%). The most common location of the lesions was the lip in 2081 cases (27.8%), followed by the gingiva in 2024 cases (27.0%), bucca in 1069 cases (14.3%), tongue in 981 cases (13.1%), and the facial skin in 695 cases (9.3%). After taking biopsy, the majority of benign lesions were treated with cryo-, laser-, or combined (cryo and laser) surgery. Conclusion: The present computer-aided study showed that irritational fibroma was the most common orofacial benign tumor, and the lip was the most frequent location. The diagnostic classification and the methodology are considerably different in the majority of the studies, which may hinder the exact comparison with other surveys from different regions of the world. Orv Hetil. 2018; 159(37): 1516–1524

Cardioprotection by remote ischemic preconditioning of the rat heart is mediated by extracellular vesicles

Remote ischemic preconditioning (RIPC) of the heart is exerted by brief ischemic insults affected on a remote organ or a remote area of the heart before a sustained cardiac ischemia. To date, little is known about the inter-organ transfer mechanisms of cardioprotection by RIPC. Exosomes and microvesicles/microparticles are vesicles of 30-100nm and 100-1000nm in diameter, respectively (collectively termed extracellular vesicles [EVs]). Their content of proteins, mRNAs and microRNAs, render EVs ideal conveyors of inter-organ communication. However, whether EVs are involved in RIPC, is unknown. Therefore, here we investigated whether (1) IPC induces release of EVs from the heart, and (2) EVs are necessary for cardioprotection by RIPC. Hearts of male Wistar rats were isolated and perfused in Langendorff mode. A group of donor hearts was exposed to 3x5-5min global ischemia and reperfusion (IPC) or 30min aerobic perfusion, while coronary perfusates were collected. Coronary perfusates of these hearts were given to another set of recipient isolated hearts. A group of recipient hearts received IPC effluent depleted of EVs by differential ultracentrifugation. Infarct size was determined after 30min global ischemia and 120min reperfusion. The presence or absence of EVs in perfusates was confirmed by dynamic light scattering, the EV marker HSP60 Western blot, and electron microscopy. IPC markedly increased EV release from the heart as assessed by HSP60. Administration of coronary perfusate from IPC donor hearts attenuated infarct size in non-preconditioned recipient hearts (12.9+/-1,6% vs. 25.0+/-2.7%), similarly to cardioprotection afforded by IPC (7.3+/-2.7% vs. 22.1+/-2.9%) on the donor hearts. Perfusates of IPC hearts depleted of EVs failed to exert cardioprotection in recipient hearts (22.0+/-2.3%). This is the first demonstration that EVs released from the heart after IPC are necessary for cardioprotection by RIPC, evidencing the importance of vesicular transfer mechanisms in remote cardioprotection

The role of the metabolite cargo of extracellular vesicles in tumor progression

Metabolomic reprogramming in tumor and stroma cells is a hallmark of cancer but understanding its effects on the metabolite composition and function of tumor-derived extracellular vesicles (EVs) is still in its infancy. EVs are membrane-bound sacs with a complex molecular composition secreted by all living cells. They are key mediators of intercellular communication both in normal and pathological conditions and play a crucial role in tumor development. Although lipids are major components of EVs, most of the EV cargo studies have targeted proteins and nucleic acids. The potential of the EV metabolome as a source for biomarker discovery has gained recognition recently, but knowledge on the biological activity of tumor EV metabolites still remains limited. Therefore, we aimed (i) to compile the list of metabolites identified in tumor EVs isolated from either clinical specimens or in vitro samples and (ii) describe their role in tumor progression through literature search and pathway analysis

Extracelluláris vezikulák és hematológiai malignitásokban játszott szerepük

Absztrakt Extracelluláris vesiculák minden szervezetben képződnek. Három legintenzívebben vizsgált csoportjuk az apoptotikus testek, a microvesiculák és az exosomák. A sejtek közötti kommunikációban, immunreakciókban, angiogenezisben betöltött szerepük csak néhány az eddig megismertek közül. A fiziológiás folyamatok mellett sokféle betegségben leírták változásaikat; a patomechanizmusban betöltött szerepük mellett felvetődik potenciális használatuk biomarkerekként. A szerzők betekintést kívánnak nyújtani az extracelluláris vesiculák kutatásába, kiemelve azt a néhány tanulmányt, amely a hematológiai malignitásokra fókuszált. A microvesiculák és exosomák vérplazmában mért mennyisége, a terápia során megfigyelt minőségi változása miatt felmerült, hogy a diagnosztikában, prognosztikában, illetve a minimális residualis betegség monitorozásában is használhatók lehetnek. Akut myeloid leukaemiában a természetes ölősejtek aktivitásának szupresszálásában bizonyított a blasteredetű exosomák szerepe. Krónikus lymphoid leukaemiában a microvesiculák közreműködése valószínű a gyógyszer-rezisztencia kialakulásában is. Orv. Hetil., 2016, 157(35), 1379–1384. | Abstract Extracellular vesicles are produced in all organisms. The most intensively investigated categories of extracellular vesicles include apoptotic bodies, microvesicles and exosomes. Among a very wide range of areas, their role has been confirmed in intercellular communication, immune response and angiogenesis (in both physiological and pathological conditions). Their alterations suggest the potential use of them as biomarkers. In this paper the authors give an insight into the research of extracellular vesicles in general, and then focus on published findings in hematological malignancies. Quantitative and qualitative changes of microvesicles and exosomes may have value in diagnostics, prognostics and minimal residual disease monitoring of hematological malignancies. The function of extracellular vesicles in downregulation of natural killer cells’ activity has been demonstrated in acute myeloid leukemia. In chronic lymphocytic leukemia, microvesicles seem to play a role in drug resistance. Orv. Hetil., 2016, 157(35), 1379–1384

MMP-9 as Prognostic Marker for Brain Tumours: A Comparative Study on Serum-Derived Small Extracellular Vesicles

Matrix metalloproteinase-9 (MMP-9) degrades the extracellular matrix, contributes to tumour cell invasion and metastasis, and its elevated level in brain tumour tissues indicates poor prognosis. High-risk tissue biopsy can be replaced by liquid biopsy; however, the blood–brain barrier (BBB) prevents tumour-associated components from entering the peripheral blood, making the development of blood-based biomarkers challenging. Therefore, we examined the MMP-9 content of small extracellular vesicles (sEVs)—which can cross the BBB and are stable in body fluids—to characterise tumours with different invasion capacity. From four patient groups (glioblastoma multiforme, brain metastases of lung cancer, meningioma, and lumbar disc herniation as controls), 222 serum-derived sEV samples were evaluated. After isolating and characterising sEVs, their MMP-9 content was measured by ELISA and assessed statistically (correlation, paired t-test, Welch’s test, ANOVA, ROC). We found that the MMP-9 content of sEVs is independent of gender and age, but is affected by surgical intervention, treatment, and recurrence. We found a relation between low MMP-9 level in sEVs (<28 ppm) and improved survival (8-month advantage) of glioblastoma patients, and MMP-9 levels showed a positive correlation with aggressiveness. These findings suggest that vesicular MMP-9 level might be a useful prognostic marker for brain tumours

High Mobility Group Box 1 Protein Induction by Mycobacterium Bovis BCG

High mobility group box 1 protein (HMGB1), a nuclear protein, is a critical cytokine that mediates the response to infection, injury, and inflammation. The aim of our study was to elaborate a reliable in vitro model to investigate whether Mycobacterium bovis BCG is able to induce HMGB1 secretion from the monocytic U-937 cells. Western blot technique was applied for the detection of HMGB1 from supernatants of cells, following induction with Mycobacterium bovis BCG. Densitometric analysis revealed higher concentrations of HMGB1 in cell supernatants stimulated with BCG than in the supernatants of the control, nonstimulated cells. Further quantitation of the secreted HMGB1 was performed by ELISA. The BCG strain resulted in a higher amount of secreted HMGB1 (450 ± 44 ng/mL) than that of LPS (84 ± 12 ng/mL) or Staphylococcus aureus (150 ± 14 ng/mL). BCG and Phorbol −12-myristate −13 acetate (PMA), added together, resulted in the highest HMGB1 secretion (645 ± 125 ng/mL). The translocation of the HMGB1 towards the cytoplasm following infection of cells with BCG was demonstrated by immunofluorescence examinations. Conclusion: Our pilot experiments draw attention to the HMGB1 inducing ability of Mycobacterium bovis. Assesment of the pathophysiological role of this late cytokine in mycobacterial infections demands further in vitro and in vivo examinations