Prominin-1 (CD133) is a cholesterol-binding membrane glycoprotein selectively associated with highly curved and prominent membrane structures. It is widely recognized as an antigenic marker of stem cells and cancer stem cells and is frequently used to isolate them from biological and clinical samples. Recent progress in understanding various aspects of CD133 biology in different cell types has revealed the involvement of CD133 in the architecture and dynamics of plasma membrane protrusions, such as microvilli and cilia, including the release of extracellular vesicles, as well as in various signaling pathways, which may be regulated in part by posttranslational modifications of CD133 and its interactions with a variety of proteins and lipids. Hence, CD133 appears to be a master regulator of cell signaling as its engagement in PI3K/Akt, Src-FAK, Wnt/β-catenin, TGF-β/Smad and MAPK/ERK pathways may explain its broad action in many cellular processes, including cell proliferation, differentiation, and migration or intercellular communication. Here, we summarize early studies on CD133, as they are essential to grasp its novel features, and describe recent evidence demonstrating that this unique molecule is involved in membrane dynamics and molecular signaling that affects various facets of tissue homeostasis and cancer development. We hope this review will provide an informative resource for future efforts to elucidate the details of CD133's molecular function in health and disease.
- Publikační typ
- časopisecké články MeSH
Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
nestr.
This detailed study examines how to overcome Pgp-mediated drug resistance in cell lines derived from various types of pediatric solid tumors by rationalized combined treatment with novel thiosemicarbazones and several types of anti-cancer drugs with lysosomotropic properties. These compounds are accumulated in the acidic pH of the lysosome. Thiosemicarbazones also enter lysosomes via Pgp, induce lysosomal permeabilization and release the stored drug from the lysosome that results in synergism. Both established model cell lines and "in-house" primary tumor cell lines will be included. An evaluation of ABC transporter expression will be performed in these cell lines followed by different types of combined treatment with selected drugs to reach the optimal treatment schedule. Intracellular distribution of the thiosemicarbazones will be analyzed using fluorescent dyes specific for various organelles to identify other intracellular sites of action. This approach represents an innovative frontier strategy how to use Pgp against itself to overcome the resistance this protein mediates.
Tato studie se detailně zabývá možností překonání lékové rezistence zprostředkované Pgp v buněčných liniích derivovaných z různých typů solidních nádorů dětského věku, a to pomocí kombinovaného působení nových thiosemicarbazonů a několika typů protinádorových léčiv s lysozomotropními vlastnostmi. Tyto látky jsou akumulovány v prostředí kyselého pH uvnitř lysozomu. Thiosemicarbazony se rovněž dostávají do lysozomů prostřednictvím Pgp, indukují permeabilizaci lysozomů a tím uvolňují látky v nich obsažené, což vede k synergickému efektu. Do projektu budou zahrnuty modelové stabilizované linie i linie derivované z tkání primárních nádorů. U těchto linií bude provedeno hodnocení exprese ABC transportérů a následně kombinované působení vybranými léčivy pro určení optimálního způsobu aplikace. Intracelulární distribuce thiosemicarbazonů bude sledována pomocí fluorescenčních barviv specifických pro různé organely, aby byla identifikována další možná místa jejich působení. Tento přístup znamená zcela inovativní strategii využití Pgp k překonání jím samotným zprostředkované lékové rezistence.
- Klíčová slova
- Léková rezistence, nízkomolekulární inhibitory, drug resistance, solidní nádory dětského věku, Pgp, lysozomy, thiosemicarbazony, antracykliny, vinca alkaloidy, pediatric solid tumors, Pgp, lysosomes, thiosemicarbazones, anthracyclines, vinca alkaloides, low-molecular inhibitors,
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR
BACKGROUND: Wilms tumor is the most common childhood kidney cancer. Two distinct histological subtypes of Wilms tumor have been described: tumors lacking anaplasia (the favorable subtype) and tumors displaying anaplastic features (the unfavorable subtype). Children with favorable disease generally have a very good prognosis, whereas those with anaplasia are oftentimes refractory to standard treatments and suffer poor outcomes, leading to an unmet clinical need. MYCN dysregulation has been associated with a number of pediatric cancers including Wilms tumor. PROCEDURES: In this context, we undertook a functional genomics approach to uncover novel therapeutic strategies for those patients with anaplastic Wilms tumor. Genomic analysis and in vitro experimentation demonstrate that cell growth can be reduced by modulating MYCN overexpression via bromodomain 4 (BRD4) inhibition in both anaplastic and nonanaplastic Wilms tumor models. RESULTS: We observed a time-dependent reduction of MYCN and MYCC protein levels upon BRD4 inhibition in Wilms tumor cell lines, which led to cell death and proliferation suppression. BRD4 inhibition significantly reduced tumor volumes in Wilms tumor patient-derived xenograft (PDX) mouse models. CONCLUSIONS: We suggest that AZD5153, a novel dual-BRD4 inhibitor, can reduce MYCN levels in both anaplastic and nonanaplastic Wilms tumor cell lines, reduces tumor volume in Wilms tumor PDXs, and should be further explored for its therapeutic potential.
- MeSH
- anaplazie genetika MeSH
- dítě MeSH
- down regulace MeSH
- jaderné proteiny genetika MeSH
- lidé MeSH
- myši MeSH
- nádory ledvin * farmakoterapie genetika metabolismus MeSH
- proteiny buněčného cyklu genetika metabolismus MeSH
- protoonkogen n-myc genetika MeSH
- transkripční faktory genetika metabolismus MeSH
- Wilmsův nádor * farmakoterapie genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- 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
Tyrosine kinase inhibitors (TKIs) are frequently used in combined therapy to enhance treatment efficacy and overcome drug resistance. The present study analyzed the effects of three inhibitors, sunitinib, gefitinib, and lapatinib, combined with iron-chelating agents, di-2-pyridylketone-4,4-dimethyl-3-thiosemicarbazone (Dp44mT) or di-2-pyridylketone-4-cyclohexyl-4-methyl-3-thiosemicarbazone (DpC). Simultaneous administration of the drugs consistently resulted in synergistic and/or additive activities against the cell lines derived from the most frequent types of pediatric solid tumors. The results of a detailed analysis of cell signaling in the neuroblastoma cell lines revealed that TKIs inhibited the phosphorylation of the corresponding receptor tyrosine kinases, and thiosemicarbazones downregulated the expression of epidermal growth factor receptor, platelet-derived growth factor receptor, and insulin-like growth factor-1 receptor, leading to a strong induction of apoptosis. Marked upregulation of the metastasis suppressor N-myc downstream regulated gene-1 (NDRG1), which is known to be activated and upregulated by thiosemicarbazones in adult cancers, was also detected in thiosemicarbazone-treated neuroblastoma cells. Importantly, these effects were more pronounced in the cells treated with drug combinations, especially with the combinations of lapatinib with thiosemicarbazones. Therefore, these results provide a rationale for novel strategies combining iron-chelating agents with TKIs in therapy of pediatric solid tumors.
- Publikační typ
- časopisecké články MeSH
Anticancer therapy by anthracyclines often leads to the development of multidrug resistance (MDR), with subsequent treatment failure. Thiosemicarbazones have been previously suggested as suitable anthracycline partners due to their ability to overcome drug resistance through dual Pgp-dependent cytotoxicity-inducing effects. Here, we focused on combining anthracyclines (doxorubicin, daunorubicin, and mitoxantrone) and two thiosemicarbazones (DpC and Dp44mT) for treating cell types derived from the most frequent pediatric solid tumors. Our results showed synergistic effects for all combinations of treatments in all tested cell types. Nevertheless, further experiments revealed that this synergism was independent of Pgp expression but rather resulted from impaired DNA repair control leading to cell death via mitotic catastrophe. The downregulation of checkpoint kinase 1 (CHEK1) expression by thiosemicarbazones and the ability of both types of agents to induce double-strand breaks in DNA may explain the Pgp-independent synergism between anthracyclines and thiosemicarbazones. Moreover, the concomitant application of these agents was found to be the most efficient approach, achieving the strongest synergistic effect with lower concentrations of these drugs. Overall, our study identified a new mechanism that offers an avenue for combining thiosemicarbazones with anthracyclines to treat tumors regardless the Pgp status.
- MeSH
- antibiotika antitumorózní MeSH
- antracykliny * farmakologie MeSH
- checkpoint kinasa 1 metabolismus MeSH
- dítě MeSH
- doxorubicin metabolismus farmakologie MeSH
- inhibitory topoisomerasy II MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- P-glykoprotein metabolismus MeSH
- poškození DNA MeSH
- thiosemikarbazony * farmakologie MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Despite constant advances in the field of pediatric oncology, the survival rate of high-risk neuroblastoma patients remains poor. The molecular and genetic features of neuroblastoma, such as MYCN amplification and stemness status, have established themselves not only as potent prognostic and predictive factors but also as intriguing targets for personalized therapy. Novel thiosemicarbazones target both total level and activity of a number of proteins involved in some of the most important signaling pathways in neuroblastoma. In this study, we found that di-2-pyridylketone 4-cyclohexyl-4-methyl-3-thiosemicarbazone (DpC) potently decreases N-MYC in MYCN-amplified and c-MYC in MYCN-nonamplified neuroblastoma cell lines. Furthermore, DpC succeeded in downregulating total EGFR and phosphorylation of its most prominent tyrosine residues through the involvement of NDRG1, a positive prognostic marker in neuroblastoma, which was markedly upregulated after thiosemicarbazone treatment. These findings could provide useful knowledge for the treatment of MYC-driven neuroblastomas that are unresponsive to conventional therapies.
- MeSH
- amplifikace genu účinky léků MeSH
- biologické modely MeSH
- chelátory železa farmakologie MeSH
- down regulace účinky léků MeSH
- erbB receptory metabolismus MeSH
- fosforylace účinky léků MeSH
- fyziologický stres účinky léků MeSH
- intracelulární signální peptidy a proteiny metabolismus MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- neuroblastom metabolismus patologie MeSH
- proteiny buněčného cyklu metabolismus MeSH
- protoonkogen n-myc metabolismus MeSH
- protoonkogenní proteiny c-akt metabolismus MeSH
- pyridiny farmakologie MeSH
- signální transdukce * MeSH
- thiosemikarbazony farmakologie MeSH
- tvar buňky účinky léků MeSH
- umlčování genů účinky léků MeSH
- upregulace účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
Giant-cell tumor of bone (GCTB) is an intermediate type of primary bone tumor characterized by locally aggressive growth with metastatic potential. The aim of this study was to identify new druggable targets among the cell signaling molecules involved in GCTB tumorigenesis. Profiles of activated signaling proteins in fresh-frozen tumor samples and tumor-derived cell lines were determined using phosphoprotein arrays. Analysis of the obtained data revealed epidermal growth factor receptor (EGFR) and platelet-derived growth factor receptor beta (PDGFRβ) as potential targets, but only the PDGFR inhibitor sunitinib caused a considerable decrease in stromal cell viability in vitro. Furthermore, in the case of a 17-year-old patient suffering from GCTB, we showed that the addition of sunitinib to the standard treatment of GCTB with the monoclonal antibody denosumab resulted in the complete depletion of multinucleated giant cells and mononuclear stromal cells in the tumor tissue. To summarize, the obtained data showed that a specific receptor tyrosine kinase (RTK) signaling pattern is activated in GCTB cells and plays an important role in the regulation of cell proliferation. Thus, activated RTKs and their downstream signaling pathways represent useful targets for precision treatment with low-molecular-weight inhibitors or with other types of modern biological therapy.
- Publikační typ
- časopisecké články MeSH
Impairment of the p53 pathway is a critical event in cancer. Therefore, reestablishing p53 activity has become one of the most appealing anticancer therapeutic strategies. Here, we disclose the p53-activating anticancer drug (3S)-6,7-bis(hydroxymethyl)-5-methyl-3-phenyl-1H,3H-pyrrolo[1,2-c]thiazole (MANIO). MANIO demonstrates a notable selectivity to the p53 pathway, activating wild-type (WT)p53 and restoring WT-like function to mutant (mut)p53 in human cancer cells. MANIO directly binds to the WT/mutp53 DNA-binding domain, enhancing the protein thermal stability, DNA-binding ability, and transcriptional activity. The high efficacy of MANIO as an anticancer agent toward cancers harboring WT/mutp53 is further demonstrated in patient-derived cells and xenograft mouse models of colorectal cancer (CRC), with no signs of undesirable side effects. MANIO synergizes with conventional chemotherapeutic drugs, and in vitro and in vivo studies predict its adequate drug-likeness and pharmacokinetic properties for a clinical candidate. As a single agent or in combination, MANIO will advance anticancer-targeted therapy, particularly benefiting CRC patients harboring distinct p53 status.
- MeSH
- antitumorózní látky chemická syntéza farmakologie MeSH
- apoptóza účinky léků genetika MeSH
- cisplatina farmakologie MeSH
- doxorubicin farmakologie MeSH
- fluorouracil farmakologie MeSH
- HCT116 buňky MeSH
- kolorektální nádory farmakoterapie genetika metabolismus patologie MeSH
- kontrolní body buněčného cyklu účinky léků genetika MeSH
- lidé MeSH
- myši nahé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádorový supresorový protein p53 agonisté genetika metabolismus MeSH
- objevování léků MeSH
- proliferace buněk účinky léků MeSH
- protokoly antitumorózní kombinované chemoterapie farmakologie MeSH
- pyrroly chemická syntéza farmakologie MeSH
- regulace genové exprese u nádorů MeSH
- synergismus léků MeSH
- thiazoly chemická syntéza farmakologie MeSH
- vazba proteinů MeSH
- xenogenní modely - testy antitumorózní aktivity MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
