Triple-negative breast cancer (TNBC) is an aggressive and complex subtype of breast cancer that lacks targeted therapy. TNBC manifests characteristic, extensive intratumoral heterogeneity that promotes disease progression and influences drug response. Single-cell techniques in combination with next-generation computation provide an unprecedented opportunity to identify molecular events with therapeutic potential. Here, we describe the generation of a comprehensive mass cytometry panel for multiparametric detection of 23 phenotypic markers and 13 signaling molecules. This single-cell proteomic approach allowed us to explore the landscape of TNBC heterogeneity, with particular emphasis on the tumor microenvironment. We prospectively profiled freshly resected tumors from 26 TNBC patients. These tumors contained phenotypically distinct subpopulations of cancer and stromal cells that were associated with the patient's clinical status at the time of surgery. We further classified the epithelial-mesenchymal plasticity of tumor cells, and molecularly defined phenotypically diverse populations of tumor-associated stroma. Furthermore, in a retrospective tissue-microarray TNBC cohort, we showed that the level of CD97 at the time of surgery has prognostic potential.

• Klíčová slova
• mass cytometry, phenotypic plasticity, single-cell profiles, triple-negative breast cancer, tumor heterogeneity, unsupervised machine learning algorithm,
• MeSH
• buňky stromatu metabolismus   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí MeSH
• proteomika MeSH
• retrospektivní studie MeSH
• signální transdukce MeSH
• triple-negativní karcinom prsu * metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Papillary thyroid carcinoma with desmoid-type fibromatosis or nodular fasciitis-like stroma is an extremely unusual and poorly understood subtype of papillary thyroid cancer. Although prior studies have demonstrated alterations in the Wnt/β-catenin signaling pathway in some of these tumors, controversy still exists regarding the nature of the stromal spindle component. We have studied seven cases of papillary thyroid carcinoma with prominent myofibroblastic stroma, including six men and one woman aged 20-65 years (mean age = 44). All cases displayed areas consistent with conventional papillary thyroid carcinoma embedded in abundant myofibroblastic-like stroma. The myofibroblastic stroma in six cases resembled desmoid-type fibromatosis and in one case it more closely resembled nodular fasciitis. By immunohistochemical staining, the stromal spindle component showed positivity for SMA and low MIB1 proliferation index in all cases, and there was at least patchy strong nuclear positivity for beta-catenin in six/seven cases. Stains for cytokeratin AE1/AE3 and PAX8 were positive in the epithelial elements but negative in the stromal component. Next-generation sequencing was performed on six of seven cases. CTNNB1 gene mutations were identified in six/seven cases. The epithelial component showed BRAF mutations in two cases and an NRAS mutation in one case. The case with fasciitis-like stroma was negative for beta-catenin by sequencing and immunostaining as well as negative for USP6 gene rearrangement. Our findings indicate that papillary thyroid carcinoma with prominent myofibroblastic stroma may represent more than one category of lesions.

• MeSH
• beta-katenin genetika   metabolismus   MeSH
• buňky stromatu metabolismus   patologie   MeSH
• dospělí MeSH
• imunohistochemie MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mutace MeSH
• myofibroblasty metabolismus   patologie   MeSH
• nádorové biomarkery metabolismus   MeSH
• nádory štítné žlázy genetika   metabolismus   patologie   MeSH
• papilární karcinom štítné žlázy genetika   metabolismus   patologie   MeSH
• protoonkogenní proteiny B-Raf genetika   metabolismus   MeSH
• senioři MeSH
• štítná žláza metabolismus   patologie   MeSH
• transkripční faktor PAX8 genetika   metabolismus   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• beta-katenin MeSH
• BRAF protein, human MeSH Prohlížeč
• CTNNB1 protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• PAX8 protein, human MeSH Prohlížeč
• protoonkogenní proteiny B-Raf MeSH
• transkripční faktor PAX8 MeSH

The similarity between humans and pigs, when it comes to tissue morphology, makes Sus scrofa not only a good research model, but also a potential source of cells for tissue engineering. Cell samples obtained from the pig donor, could be influenced in vitro, in order to become a source of tissue material for xenotransplantation, reconstructive and regenerative medicine. Significant amounts of data point to especially major similarities in pig and human reproductive systems. Because of that, particular scientific focus is centered on research concerning porcine COCs, theca and granulosa cells in primary cultures. One of the aspects of the reproductive process, that is still largely undiscovered, is the interaction between preimplantation blastocyst and maternal uterine tissues. In this study, we used molecular analysis techniques, such as RT-qPCR and immunocytochemistry, to analyze the expression and distribution of cytokeratin 18 and panCytokeratins 8, 18 and 19 and vimentin in porcine luminal endometrial epithelial cells, coupled with analysis of their behavior in RTCA. The results have confirmed the presence of epithelial, as well as stromal cell markers in the cells, varying in levels at different stages of culture. They have also given insight into the modes of proliferation and differentiation of studied cells in in vitro culture, as well as providing additional proof for the possible mesenchymal transdifferentiation of epithelial cells.

• MeSH
• biologické markery metabolismus   MeSH
• biologické modely MeSH
• buněčná diferenciace MeSH
• buňky stromatu cytologie   metabolismus   MeSH
• časové faktory MeSH
• endometrium cytologie   MeSH
• epitelové buňky cytologie   metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• modely u zvířat MeSH
• prasata MeSH
• proliferace buněk * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH

Human tumor xenografts in mice together with the species-specific analysis of expressed genes allow to study the molecular processes driving tumor growth and progression in vivo and help to develop and evaluate anticancer therapies. In the present work, we designed and validated species-specific real-time RT-PCR assays for discrimination and quantitation of expression of human and mouse transcripts in cancer and stromal cells including dipeptidyl peptidase (DPP) 4, DPP8, DPP9, fibroblast activation protein (FAP) and CXC chemokine receptor 4 in mixed human-mouse biological samples. Using single species RNA samples and mixed human-mouse RNA samples, we formulated and characterized two-step real-time RT-PCR assays to quantitate expression of the indicated transcripts and described analytical performance of the assays. We also demonstrated the applicability of these assays for species-specific quantitation of transcriptional expression of mouse stromal cell genes including Dpp4, Dpp8, Dpp9, Fap and Cxcr4 in mixed human-mouse RNA samples from human glioma cell-derived tumor xenografts growing in mouse brain.

• Klíčová slova
• CXCR4, Dipeptidyl peptidase, Fibroblast activation protein, Glioma, Species-specific expression, Tumor stroma,
• MeSH
• buňky stromatu metabolismus   MeSH
• dipeptidylpeptidasy a tripeptidylpeptidasy metabolismus   MeSH
• druhová specificita MeSH
• gliom metabolismus   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• mozek metabolismus   MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory mozku metabolismus   MeSH
• proteom metabolismus   MeSH
• receptory CXCR4 metabolismus   MeSH
• regulace genové exprese u nádorů * MeSH
• stanovení celkové genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dipeptidylpeptidasy a tripeptidylpeptidasy MeSH
• proteom MeSH
• receptory CXCR4 MeSH

Adipose/fat tissue provides an abundant source of stromal vascular fraction (SVF) cells for immediate administration and can also give rise to a substantial number of cultured, multipotent adipose-derived stromal cells (ADSCs). Recently, both SVF and ADSCs have gained wide-ranging translational significance in regenerative medicine. Initially used for cosmetic breast enhancement, this mode of treatment has found use in many diseases involving immune disorders, tissue degeneration, and ischaemic conditions. In this review, we try to address several important aspects of this field, outlining the biology, technology, translation, and challenges related to SVF- and ADSC-based therapies. Starting from the basics of SVF and ADSC isolation, we touch upon recently developed technologies, addressing elements of novel methods and devices under development for point-of-care isolation of SVF. Characterisation of SVF cells and ADSCs is also an evolving area and we look into unusual expression of CD34 antigen as an interesting marker for such purposes. Based on reports involving different cells of the SVF, we draw a potential mode of action, focussing on angiogenesis since it involves multiple cells, unlike immunomodulation which is governed predominantly by ADSCs. We have looked into the latest research, experimental therapies, and clinical trials which are utilising SVF/ADSCs in conditions such as multiple sclerosis, Crohn's disease, peripheral neuropathy, osteoarthritis, diabetic foot ulcer, and so forth. However, problems have arisen with regards to the lack of proper regulatory guidelines for such therapies and, since the introduction of US Food and Drug Administration draft guidelines and the Reliable and Effective Growth for Regenerative Health Options that Improve Wellness (REGROW) Act, the debate became more public with regards to safe and efficacious use of these cells.

• Klíčová slova
• CD34, Multipotent-stromal cells, Point-of-care biomedical devices, Regenerative medicine, Regulation of stem cell therapeutics, Stromal vascular fraction,
• MeSH
• buňky stromatu metabolismus   patologie   transplantace   MeSH
• Crohnova nemoc metabolismus   patologie   terapie   MeSH
• diabetická noha metabolismus   patologie   terapie   MeSH
• lidé MeSH
• multipotentní kmenové buňky metabolismus   patologie   transplantace   MeSH
• nemoci periferního nervového systému metabolismus   patologie   terapie   MeSH
• osteoartróza metabolismus   patologie   terapie   MeSH
• regenerativní lékařství metody   MeSH
• roztroušená skleróza metabolismus   patologie   terapie   MeSH
• tuková tkáň metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Malignant progression is greatly affected by dynamic cross-talk between stromal and cancer cells. Exosomes are secreted nanovesicles that have key roles in cell-cell communication by transferring nucleic acids and proteins to target cells and tissues. Recently, MicroRNAs (miRs) and their delivery in exosomes have been implicated in physiological and pathological processes. Tumor-delivered miRs, interacting with stromal cells in the tumor microenvironment, modulate tumor progression, angiogenesis, metastasis and immune escape. Altered cell metabolism is one of the hallmarks of cancer. A number of different types of tumor rely on mitochondrial metabolism by triggering adaptive mechanisms to optimize their oxidative phosphorylation in relation to their substrate supply and energy demands. Exogenous exosomes can induce metabolic reprogramming by restoring the respiration of cancer cells and supress tumor growth. The exosomal miRs involved in the modulation of cancer metabolism may be potentially utilized for better diagnostics and therapy.

• MeSH
• biologický transport MeSH
• buňky stromatu metabolismus   MeSH
• energetický metabolismus MeSH
• exozómy genetika   MeSH
• genetická terapie MeSH
• lidé MeSH
• mezibuněčná komunikace MeSH
• mikro RNA genetika   MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí genetika   MeSH
• nádory genetika   metabolismus   patologie   terapie   MeSH
• regulace genové exprese u nádorů MeSH
• signální transdukce MeSH
• technika přenosu genů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• mikro RNA MeSH

Glioblastomas are deadly neoplasms resistant to current treatment modalities. Fibroblast activation protein (FAP) is a protease which is not expressed in most of the normal adult tissues but is characteristically present in the stroma of extracranial malignancies. FAP is considered a potential therapeutic target and is associated with a worse patient outcome in some cancers. The FAP localization in the glioma microenvironment and its relation to patient survival are unknown. By analyzing 56 gliomas and 15 non-tumorous brain samples, we demonstrate increased FAP expression in a subgroup of high-grade gliomas, in particular on the protein level. FAP expression was most elevated in the mesenchymal subtype of glioblastoma. It was neither associated with glioblastoma patient survival in our patient cohort nor in publicly available datasets. FAP was expressed in both transformed and stromal cells; the latter were frequently localized around dysplastic blood vessels and commonly expressed mesenchymal markers. In a mouse xenotransplantation model, FAP was expressed in glioma cells in a subgroup of tumors that typically did not express the astrocytic marker GFAP. Endogenous FAP was frequently upregulated and part of the FAP+ host cells coexpressed the CXCR4 chemokine receptor. In summary, FAP is expressed by several constituents of the glioblastoma microenvironment, including stromal non-malignant mesenchymal cells recruited to and/or activated in response to glioma growth. The limited expression of FAP in healthy tissues together with its presence in both transformed and stromal cells suggests that FAP may be a candidate target for specific delivery of therapeutic agents in glioblastoma.

• Klíčová slova
• Fibroblast activation protein α, Glioma, Seprase, Serine protease, Stromal cells,
• MeSH
• apoptóza MeSH
• buňky stromatu metabolismus   patologie   MeSH
• dospělí MeSH
• endopeptidasy MeSH
• fibroblasty metabolismus   patologie   MeSH
• glioblastom genetika   metabolismus   patologie   MeSH
• imunoenzymatické techniky MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové proteiny genetika   metabolismus   MeSH
• messenger RNA genetika   MeSH
• mezoderm metabolismus   patologie   MeSH
• míra přežití MeSH
• myši inbrední NOD MeSH
• myši MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorové buňky kultivované MeSH
• následné studie MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• prognóza MeSH
• proliferace buněk MeSH
• senioři MeSH
• serinové endopeptidasy genetika   metabolismus   MeSH
• staging nádorů MeSH
• studie případů a kontrol MeSH
• transformované buněčné linie metabolismus   patologie   MeSH
• western blotting MeSH
• xenogenní modely - testy antitumorózní aktivity MeSH
• želatinasy genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• senioři MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• endopeptidasy MeSH
• fibroblast activation protein alpha MeSH Prohlížeč
• membránové proteiny MeSH
• messenger RNA MeSH
• nádorové biomarkery MeSH
• serinové endopeptidasy MeSH
• želatinasy MeSH

The role of the local microenvironment in influencing cell behavior is central to both normal development and cancer formation. Here, we show that sprouty 1 (SPRY1) modulates the microenvironment to enable proper mammary branching morphogenesis. This process occurs through negative regulation of epidermal growth factor receptor (EGFR) signaling in mammary stroma. Loss of SPRY1 resulted in up-regulation of EGFR-extracellular signal-regulated kinase (ERK) signaling in response to amphiregulin and transforming growth factor alpha stimulation. Consequently, stromal paracrine signaling and ECM remodeling is augmented, leading to increased epithelial branching in the mutant gland. By contrast, down-regulation of EGFR-ERK signaling due to gain of Sprouty function in the stroma led to stunted epithelial branching. Taken together, our results show that modulation of stromal paracrine signaling and ECM remodeling by SPRY1 regulates mammary epithelial morphogenesis during postnatal development.

• Klíčová slova
• EGF signaling, FGF signaling, branching morphogenesis, epithelial–stromal interactions, stromal microenvironment,
• MeSH
• adaptorové proteiny signální transdukční nedostatek   metabolismus   MeSH
• amfiregulin farmakologie   MeSH
• buňky stromatu účinky léků   metabolismus   MeSH
• časosběrné zobrazování MeSH
• epitel růst a vývoj   metabolismus   MeSH
• epitelové buňky cytologie   účinky léků   MeSH
• erbB receptory metabolismus   MeSH
• extracelulární matrix metabolismus   MeSH
• extracelulárním signálem regulované MAP kinasy metabolismus   MeSH
• fibroblasty účinky léků   metabolismus   MeSH
• fosfoproteiny nedostatek   metabolismus   MeSH
• fosforylace účinky léků   MeSH
• kolagen metabolismus   MeSH
• ligandy MeSH
• membránové proteiny nedostatek   metabolismus   MeSH
• mléčné žlázy zvířat účinky léků   metabolismus   MeSH
• morfogeneze * účinky léků   MeSH
• mutace genetika   MeSH
• myši knockoutované MeSH
• myši nahé MeSH
• parakrinní signalizace * účinky léků   MeSH
• pohyb buněk účinky léků   MeSH
• protoonkogenní proteiny c-akt metabolismus   MeSH
• signální transdukce * účinky léků   MeSH
• transformující růstový faktor alfa farmakologie   MeSH
• vývojová regulace genové exprese účinky léků   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• amfiregulin MeSH
• erbB receptory MeSH
• extracelulárním signálem regulované MAP kinasy MeSH
• fosfoproteiny MeSH
• kolagen MeSH
• ligandy MeSH
• membránové proteiny MeSH
• protoonkogenní proteiny c-akt MeSH
• Spry1 protein, mouse MeSH Prohlížeč
• transformující růstový faktor alfa MeSH

Adherent, fibroblastic cells from different tissues are thought to contain subsets of tissue-specific stem/progenitor cells (often called mesenchymal stem cells). These cells display similar cell surface characteristics based on their fibroblastic nature, but also exhibit differences in molecular phenotype, growth rate, and their ability to differentiate into various cell phenotypes. The mechanisms underlying these differences remain poorly understood. We analyzed Ca(2+) signals and membrane properties in rat adipose-derived stromal cells (ADSCs) and bone marrow stromal cells (BMSCs) in basal conditions, and then following a switch into medium that contains factors known to modify their character. Modified ADSCs (mADSCs) expressed L-type Ca(2+) channels whereas both L- and P/Q- channels were operational in mBMSCs. Both mADSCs and mBMSCs possessed functional endoplasmic reticulum Ca(2+) stores, expressed ryanodine receptor-1 and -3, and exhibited spontaneous [Ca(2+)]i oscillations. The mBMSCs expressed P2X7 purinoceptors; the mADSCs expressed both P2X (but not P2X7) and P2Y (but not P2Y1) receptors. Both types of stromal cells exhibited [Ca(2+)]i responses to vasopressin (AVP) and expressed V1 type receptors. Functional oxytocin (OT) receptors were, in contrast, expressed only in modified ADSCs and BMSCs. AVP and OT-induced [Ca(2+)]i responses were dose-dependent and were blocked by their respective specific receptor antagonists. Electrophysiological data revealed that passive ion currents dominated the membrane conductance in ADSCs and BMSCs. Medium modification led to a significant shift in the reversal potential of passive currents from -40 to -50mV in cells in basal to -80mV in modified cells. Hence membrane conductance was mediated by non-selective channels in cells in basal conditions, whereas in modified medium conditions, it was associated with K(+)-selective channels. Our results indicate that modification of ADSCs and BMSCs by alteration in medium formulation is associated with significant changes in their Ca(2+) signaling and membrane properties.

• Klíčová slova
• Adipose derived stromal cells, Bone marrow stromal cell, Ca(2+) signaling, Ion channels, Ionotropic receptors, Oxytocin, Patch-clamp, Purinergic signaling, Ryanodine receptors, Vasopressin,
• MeSH
• buňky kostní dřeně cytologie   MeSH
• buňky stromatu cytologie   účinky léků   metabolismus   MeSH
• evokované potenciály účinky léků   MeSH
• glutamátové receptory metabolismus   MeSH
• iontové kanály metabolismus   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• metoda terčíkového zámku MeSH
• oxytocin farmakologie   MeSH
• potkani Sprague-Dawley MeSH
• purinergní receptory metabolismus   MeSH
• tuková tkáň cytologie   MeSH
• vápník metabolismus   MeSH
• vápníkové kanály metabolismus   MeSH
• vasopresiny farmakologie   MeSH
• videomikroskopie MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glutamátové receptory MeSH
• iontové kanály MeSH
• oxytocin MeSH
• purinergní receptory MeSH
• vápník MeSH
• vápníkové kanály MeSH
• vasopresiny MeSH

UNLABELLED: Transgelin and transgelin-2 have been discussed as potential markers of various cancers. Here we identified increased transgelin level in lymph node positive vs. negative, low grade primary breast cancer tissues using 2-DE in the cohort of 12 patients. We further clinically validated 2-DE results in an independent cohort of 48 low grade breast cancer patients through untargeted and targeted proteomics analysis (iTRAQ-2D-LC-MS/MS, mTRAQ-SRM), at transcript level and using immunohistochemistry. Another group of 48 high grade tumors of different breast cancer subtypes was analyzed together with the low grade samples to test transgelin specificity for low grade tumors and to study transgelin relation to known molecular markers and histological features. The results confirmed transgelin connection with the lymph node metastasis. As a marker of a reactive tumor stroma, transgelin can be connected with the higher risk of metastasis development. Moreover, we observed significant down-regulation of transgelin in high vs. low grade tumors caused by decreased content of stromal cells (mainly expressing transgelin) in high grade tumor tissue. We also analyzed expression of transgelin-2 in the second cohort using proteomics and immunohistochemistry. Transgelin-2 was mainly expressed by epithelial cancer cells and its levels were increased in metastatic and poorly differentiated tumors. BIOLOGICAL SIGNIFICANCE: Both transgelin and transgelin-2 have been previously described as potential markers of many types of cancer. We are specifying this connection to metastatic affection of lymph nodes and cell differentiation in breast cancer. In the wider context, the results of our study highlight tumor stroma as a source of cancer biomarkers and point out how measured levels of tissue markers can actually reflect cellular feature of cancer mass.

• Klíčová slova
• Breast cancer, Lymph node metastasis, Proteomics, Transgelin, Transgelin-2,
• MeSH
• buňky stromatu metabolismus   patologie   MeSH
• lidé MeSH
• lymfatické uzliny metabolismus   patologie   MeSH
• mikrofilamentové proteiny metabolismus   MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorové proteiny metabolismus   MeSH
• nádory prsu metabolismus   patologie   MeSH
• pilotní projekty MeSH
• regulace genové exprese u nádorů MeSH
• stupeň nádoru MeSH
• svalové proteiny metabolismus   MeSH
• upregulace MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• mikrofilamentové proteiny MeSH
• nádorové biomarkery MeSH
• nádorové proteiny MeSH
• svalové proteiny MeSH
• transgelin MeSH Prohlížeč