The protease caspase-3 is a key mediator of apoptotic programmed cell death. But weak or transient caspase activity can contribute to neuronal differentiation, axonal pathfinding, and synaptic long-term depression. Despite the importance of sublethal, or nonapoptotic, caspase activity in neurodevelopment and neural plasticity, there has been no simple method for mapping and quantifying nonapoptotic caspase activity (NACA) in rodent brains. We therefore generated a transgenic mouse expressing a highly sensitive and specific fluorescent reporter of caspase activity, with peak signal localized to the nucleus. As a proof of concept, we first obtained evidence that NACA influences neurophysiology in an amygdalar circuit. Then focusing on the amygdala, we were able to quantify a sex-specific persistent elevation in caspase activity in females after restraint stress. This simple in vivo caspase activity reporter will facilitate systems-level studies of apoptotic and nonapoptotic phenomena in behavioral and pathologic models.
- MeSH
- apoptóza * fyziologie MeSH
- kaspasa 9 MeSH
- mozek * MeSH
- myši transgenní MeSH
- myši MeSH
- neuroplasticita MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
Embryonic stem cells and induced pluripotent stem cells provided us with fascinating new knowledge in recent years. Mechanistic insight into intricate regulatory circuitry governing pluripotency stemness and disclosing parallels between pluripotency stemness and cancer instigated numerous studies focusing on roles of pluripotency transcription factors, including Oct4, Sox2, Klf4, Nanog, Sall4 and Tfcp2L1, in cancer. Although generally well substantiated as tumour-promoting factors, oncogenic roles of pluripotency transcription factors and their clinical impacts are revealing themselves as increasingly complex. In certain tumours, both Oct4 and Sox2 behave as genuine oncogenes, and reporter genes driven by composite regulatory elements jointly recognized by both the factors can identify stem-like cells in a proportion of tumours. On the other hand, cancer stem cells seem to be biologically very heterogeneous both among different tumour types and among and even within individual tumours. Pluripotency transcription factors are certainly implicated in cancer stemness, but do not seem to encompass its entire spectrum. Certain cancer stem cells maintain their stemness by biological mechanisms completely different from pluripotency stemness, sometimes even by engaging signalling pathways that promote differentiation of pluripotent stem cells. Moreover, while these signalling pathways may well be antithetical to stemness in pluripotent stem cells, they may cooperate with pluripotency factors in cancer stem cells - a paradigmatic example is provided by the MAPK-AP-1 pathway. Unexpectedly, forced expression of pluripotency transcription factors in cancer cells frequently results in loss of their tumour-initiating ability, their phenotypic reversion and partial epigenetic normalization. Besides the very different signalling contexts operating in pluripotent and cancer stem cells, respectively, the pronounced dose dependency of reprogramming pluripotency factors may also contribute to the frequent loss of tumorigenicity observed in induced pluripotent cancer cells. Finally, contradictory cell-autonomous and non-cell-autonomous effects of various signalling molecules operate during pluripotency (cancer) reprogramming. The effects of pluripotency transcription factors in cancer are thus best explained within the concept of cancer stem cell heterogeneity.
- MeSH
- buněčná diferenciace genetika MeSH
- embryonální kmenové buňky MeSH
- indukované pluripotentní kmenové buňky * metabolismus MeSH
- lidé MeSH
- nádory * genetika metabolismus MeSH
- oktamerní transkripční faktor 3 genetika metabolismus MeSH
- pluripotentní kmenové buňky * MeSH
- přeprogramování buněk genetika MeSH
- transkripční faktory genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
Sarcomas are a heterogeneous group of mesenchymal tumours, with a great variability in their clinical behaviour. While our knowledge of sarcoma initiation has advanced rapidly in recent years, relatively little is known about mechanisms of sarcoma progression. JUN-murine fibrosarcoma progression series consists of four sarcoma cell lines, JUN-1, JUN-2, JUN-2fos-3, and JUN-3. JUN-1 and -2 were established from a single tumour initiated in a H2K/v-jun transgenic mouse, JUN-3 originates from a different tumour in the same animal, and JUN-2fos-3 results from a targeted in vitro transformation of the JUN-2 cell line. The JUN-1, -2, and -3 cell lines represent a linear progression from the least transformed JUN-2 to the most transformed JUN-3, with regard to all the transformation characteristics studied, while the JUN-2fos-3 cell line exhibits a unique transformation mode, with little deregulation of cell growth and proliferation, but pronounced motility and invasiveness. The invasive sarcoma sublines JUN-2fos-3 and JUN-3 show complex metabolic profiles, with activation of both mitochondrial oxidative phosphorylation and glycolysis and a significant increase in spared respiratory capacity. The specific transcriptomic profile of invasive sublines features very complex biological relationships across the identified genes and proteins, with accentuated autocrine control of motility and angiogenesis. Pharmacologic inhibition of one of the autocrine motility factors identified, Ccl8, significantly diminished both motility and invasiveness of the highly transformed fibrosarcoma cell. This progression series could be greatly valuable for deciphering crucial aspects of sarcoma progression and defining new prognostic markers and potential therapeutic targets.
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
Ve dnech 14.–17. listopadu 2018 se v Římě uskutečnila velká mezinárodní konference Connective Tissue Oncology Group Annual Meeting 2018 (CTOS 2018), která svedla dohromady naprostou většinu nejvýznamnějších specialistů jak na molekulární biologii, tak i klinickou onkologii sarkomů, především sarkomů měkkých tkání. Ve dnech 8.–10. května 2019 se potom v Bergenu konala regionální konference The 39th Plenary Meeting of the Scandinavian Sarcoma Group (SSGM 2019). Soubory otázek, které byly na obou konferencích diskutovány formou přednáškových či posterových sdělení, lze rozdělit do několika zastřešujících okruhů. Byl představen velký projekt zaměřený na objasnění germinální genetiky sarkomů; dosavadní výsledky ukazují na překvapivě velký význam genů kódujících proteiny, které zajišťují integritu telomer, jakož i existenci komplexních mechanizmů genetické predispozice zahrnující polygenní dědičnost či modifikační geny. Problematice somatické genetiky sarkomů dominuje analýza vzniku a mechanizmu účinku fúzních onkogenů odpovídajících za iniciaci značné části především pediatrických sarkomů. U karyotypicky komplexních sarkomů je evidentní snaha po patobiologické specifikaci do subtypů, ať už na základě specifické klinické charakterizace (uterinní leiomyosarkom vs. leiomyosarkom měkkých tkání), či specifických expresních profilů genů (molekulární subtypy nediferencovaného pleiomorfního sarkomu). Molekulární charakterizace může být vodítkem pro koncipování subtypově specifických terapeutických protokolů, jak je tomu v různém měřítku u obou shora zmíněných příkladů. Mezi další prominentní typy sarkomů, u nichž se podařilo transformovat základní molekulárně biologické poznatky do podoby úspěšné cílené terapie, patří např. gastrointestinální stromální tumor, infantilní fibrosarkom či inflamatorní myofibroblastický tumor, a rovněž u světlobuněčného sarkomu a dediferencovaného liposarkomu byly obdobné koncepty prezentovány, byť jejich dosavadní účinnost zůstává za očekáváním, a podobná situace zatím přetrvává u osteosarkomu. Z hlediska molekulárních struktur, na které lze zaměřit cílenou terapii, zaujímá v oblasti sarkomů měkkých tkání zcela zásadní postavení signální systém destičkového růstového faktoru, a to jak u vzácných typů založených na přímé iniciační mutační aktivaci (relativně malá část gastrointestinálních stromálních tumorů, infantilní hereditární myofibromatóza, dermatofibrosarcoma protuberans), u kterých cílená terapie využívá především nízkomolekulárních tyrozinkinázových inhibitorů, tak rovněž u širokého spektra obvyklých typů sarkomů, kde se součástí kombinační chemoterapie stává specifická blokující anti-PDGFRα-protilátka olaratumab. V oblasti klinické prognózy byl pozoruhodný vývoj zaznamenán zejména u tzv. prognostických nomogramů. Zajímavé výsledky byly rovněž prezentovány v oblasti odvození nových experimentálních modelů vzniku a progrese sarkomů.
The Connective Tissue Oncology Group Annual Meeting 2018 CTOS 2018) took place in Rome from 4 to 17 November 2018, and the 39th Plenary Meeting of the Scandinavian Sarcoma Group (SSGM 2019) was held in Bergen from 8 to 10 May 2019. These two large international conferences brought together an overwhelming majority of molecular and clinical specialists in the sarcoma field, especially those working on soft tissue sarcoma. Topics discussed on the conferences included, among others, sarcoma genetics, clinical and molecular subclassification, targeted therapy, clinical prognostication, and new experimental sarcoma models. A large ongoing international study on germinal sarcoma genetics was presented, the interim results of which revealed the extremely complex nature of genetic disposition to sarcoma, and, surprisingly, a rather prominent place among predisposing genes for those coding for structural telomere constituents. Fusion oncogenes dominate somatic sarcoma genetics, especially because of their origin and impact on sarcoma clinical behaviour, and are especially relevant for karyotypically simple paediatric sarcomas. A crucial issue in karyotypically complex sarcomas are the efforts being made to obtain a subclassification of sarcoma, other than those based on pathology, using either the clinical characteristics of sarcomas (uterine leiomyosarcoma vs. soft tissue leiomyosarcoma) or specific gene expression profiles (molecular subtypes in undifferentiated pleiomorphic sarcoma), which showed that molecular characterization can open the way for subtype specific therapies. Other examples of where this type of strategy can be applied include gastrointestinal stromal tumours, infantile fibrosarcoma, and inflammatory myofibroblastic tumours, where targeted therapy could be conceived based on the actionable mutations identified. Attempts in this direction have been made also for clear cell sarcoma and dedifferentiated liposarcoma, albeit the effectiveness of molecular-targeted treatments for these sarcomas is still poor, and progress in the treatment of osteosarcoma is still rather slow. Actually, the platelet-derived growth factor signalling system holds a prominent position in searches for targeted therapies, not only against rare sarcoma types, where are activated by mutations (some gastrointestinal stromal tumours, infantile hereditary myofibromatosis, and dermatofibrosarcoma protuberans), but also against other more usual sarcoma types, where the blocking anti-PDGFRα-antibody olaratumab has been successfully integrated into combinatorial chemotherapeutic regimens. In the field of clinical prognostication, remarkable progress in sarcoma nomograms was reported. Interesting results were also presented in the area of new experimental sarcoma models.
- Klíčová slova
- cílená terapie, prognostické nomogramy, experimentální modely sarkomatogeneze,
- MeSH
- genetická predispozice k nemoci MeSH
- lidé MeSH
- osteosarkom genetika terapie MeSH
- sarkom * genetika terapie MeSH
- Check Tag
- lidé MeSH
Little attention was given to the interaction between tumor and stromal cells in urothelial bladder carcinoma (UBC). While recent studies point towards the existence of different fibroblast subsets, no comprehensive analyses linking different fibroblast markers to UBC patient survival have been performed so far. Through immunohistochemical analysis of five selected fibroblast markers, namely alpha smooth muscle actin (ASMA), CD90/Thy-1, fibroblast activation protein (FAP), platelet derived growth factor receptor-alpha and -beta (PDGFRa,-b), this study investigates their association with survival and histopathological characteristics in a cohort of 344 UBC patients, involving both, muscle-invasive and non-muscle-invasive cases. The data indicates that combinations of stromal markers are more suited to identify prognostic patient subgroups than single marker analysis. Refined stroma-marker-based patient stratification was achieved through cluster analysis and identified a FAP-dominant patient cluster as independent marker for shorter 5-year-survival (HR(95% CI)2.25(1.08-4.67), p = 0.030). Analyses of interactions between fibroblast and CD8a-status identified a potential minority of cases with CD90-defined stroma and high CD8a infiltration showing a good prognosis of more than 80% 5-year-survival. Presented analyses point towards the existence of different stroma-cell subgroups with distinct tumor-modulatory properties and motivate further studies aiming to better understand the molecular tumor-stroma crosstalk in UBC.
- MeSH
- aktiny metabolismus MeSH
- antigeny Thy-1 metabolismus MeSH
- CD8-pozitivní T-lymfocyty cytologie imunologie metabolismus MeSH
- fenotyp MeSH
- fibroblasty cytologie metabolismus MeSH
- Kaplanův-Meierův odhad MeSH
- lidé MeSH
- membránové proteiny metabolismus MeSH
- nádorové biomarkery metabolismus MeSH
- nádory močového měchýře metabolismus mortalita patologie MeSH
- prognóza MeSH
- proporcionální rizikové modely MeSH
- růstový faktor odvozený z trombocytů - receptor alfa metabolismus MeSH
- senioři MeSH
- serinové endopeptidasy metabolismus MeSH
- shluková analýza MeSH
- želatinasy metabolismus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- fyziologie výchova MeSH
- genetika výchova MeSH
- hodnocení programu MeSH
- kurikulum MeSH
- lidé MeSH
- mutace * MeSH
- pochopení MeSH
- rozvoj plánování MeSH
- studenti lékařství * psychologie MeSH
- studium lékařství pregraduální metody MeSH
- stupeň vzdělání MeSH
- učení MeSH
- vyučování * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Ovarian carcinoma features pronounced clinical, histopathological, and molecular heterogeneity. There is good reason to believe that parts of this heterogeneity can be explained by differences in the respective cell of origin, with a self-renewing fallopian tube secretory cell being likely responsible for initiation of an overwhelming majority of high-grade serous ovarian carcinomas (i.e., type II tumors according to the recent dualistic classification), whereas there are several mutually non-exclusive possibilities for the initiation of type I tumors, including ovarian surface epithelium stem cells, endometrial cells, or even cells of extra-Müllerian origin. Interestingly, both fallopian tube self-renewing secretory cells and ovarian surface epithelium stem cells seem to be characterized by an overlapping array of stemness signaling pathways, especially Wnt/β-catenin. Apart from this variability in the respective cell of origin, the particular clinical behavior of ovarian carcinoma strongly suggests an underlying stem cell component with a crucial impact. This becomes especially evident in high-grade serous ovarian carcinomas treated with classical chemotherapy, which entails a gradual evolution of chemoresistant disease without any apparent selection of clones carrying obvious chemoresistance-associated mutations. Several cell surface markers (e.g., CD24, CD44, CD117, CD133, and ROR1) as well as functional approaches (ALDEFLUOR™ and side population assays) have been used to identify and characterize putative ovarian carcinoma stem cells. We have recently shown that side population cells exhibit marked heterogeneity on their own, which can hamper their straightforward therapeutic targeting. An alternative strategy for stemness-depleting interventions is to target the stem cell niche, i.e., the specific microanatomical structure that secures stem cell maintenance and survival through provision of a set of stem cell-promoting and differentiation-antagonizing factors. Besides identifying direct or indirect therapeutic targets, profiling of side population cells and other ovarian carcinoma stem cell subpopulations can reveal relevant prognostic markers, as exemplified by our recent discovery of the Vav3.1 transcript variant, which filters out a fraction of prognostically unfavorable ovarian carcinoma cases.
