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BACKGROUND: Treatment of advanced cutaneous melanoma remains challenging, and new data on melanoma biology are required. The most widely accepted criteria for the prognostic evaluation of melanoma are histopathological and clinical parameters, and the identification of additional tumor markers is thus of paramount importance. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI), an important tool in cancer research, is useful for unraveling the molecular profile of melanoma. METHODOLOGY/PRINCIPAL FINDINGS: In this report, we used the melanoma-bearing Libechov minipig (MeLiM), a unique animal model that allows observation of the complete spontaneous regression of invasive cutaneous melanoma, to investigate i) the differences between melanoma and healthy skin protein profiles and ii) the proteins potentially involved in spontaneous regression. The MeLiM tissues were cryosected, histologically characterized, analyzed by MALDI MSI, and immunohistologically stained. Multivariate statistical analyses of the MALDI MSI data revealed ten relevant m/z ions, of which the expression levels varied significantly among the studied MeLiM tissues. These ion peaks were used to create mass ion images/maps and visualize the differences between tumor and healthy skin specimens, as well as among histologically characterized tissue regions. CONCLUSIONS/SIGNIFICANCE: Protein profiles comprising ten statistically significant mass ion peaks useful for differentiating cutaneous melanoma and healthy skin tissues were determined. Peaks at m/z 3044, 6011, 6140 and 10180 were overexpressed in melanoma compared with healthy skin tissue. More specifically, m/z 6140 was expressed at significantly (p < 0.05) higher levels in normally growing melanoma regions than in regions with early and late spontaneous regression. This study demonstrates the clinical utility of MALDI MSI for the analysis of tissue cryosections at a molecular level.
Here we developed a powerful tool for comprehensive data collection and mapping of molecular and elemental signatures in the Melanoma-bearing Libechov Minipig (MeLiM) model. The combination of different mass spectrometric methods allowed for detail investigation of specific melanoma markers and elements and their spatial distribution in tissue sections. MALDI-MSI combined with HPLC-MS/MS analyses resulted in identification of seven specific proteins, S100A12, CD163, MMP-2, galectin-1, tenascin, resistin and PCNA that were presented in the melanoma signatures. Furthermore, the ICP-MS method allowed for spatial detection of zinc, calcium, copper, and iron elements linked with the allocation of the specific binding proteins.
National cancer databases document that melanoma is the most aggressive and deadly cutaneous malignancy with worldwide increasing incidence in the Caucasian population. Around 10% of melanomas occur in families. Several germline mutations were identified that might help to indicate individuals at risk for preventive interventions and early disease detection. More than 50% of sporadic melanomas carry mutations in Ras/Raf/mitogen-activated protein kinase (MAPK/MEK) pathway, which may represent aims of novel targeted therapies. Despite advances in targeted therapies and immunotherapies, the outcomes in metastatic tumor are still unsatisfactory. Here, we review animal models that help our understanding of melanoma development and treatment, including non-vertebrate, mouse, swine, and other mammal models, with an emphasis on those with spontaneously developing melanoma. Special attention is paid to the melanoma-bearing Libechov minipig (MeLiM). This original swine model of hereditary metastatic melanoma enables studying biological processes underlying melanoma progression, as well as spontaneous regression. Current histological, immunohistochemical, biochemical, genetic, hematological, immunological, and skin microbiome findings in the MeLiM model are summarized, together with development of new therapeutic approaches based on tumor devitalization. The ongoing study of molecular and immunological base of spontaneous regression in MeLiM model has potential to bring new knowledge of clinical importance.
- MeSH
- melanom genetika MeSH
- miniaturní prasata genetika MeSH
- modely nemocí na zvířatech MeSH
- nádory kůže genetika MeSH
- prasata genetika MeSH
- progrese nemoci MeSH
- sekundární malignity genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Publikační typ
- abstrakt z konference MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- gama-glutamyltransferasa analýza krev MeSH
- mannosidasy analýza krev MeSH
- melanom experimentální enzymologie MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- kongresy MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- melanom experimentální imunologie terapie MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- kongresy MeSH
UNLABELLED: We previously showed that close relatives of human coronavirus 229E (HCoV-229E) exist in African bats. The small sample and limited genomic characterizations have prevented further analyses so far. Here, we tested 2,087 fecal specimens from 11 bat species sampled in Ghana for HCoV-229E-related viruses by reverse transcription-PCR (RT-PCR). Only hipposiderid bats tested positive. To compare the genetic diversity of bat viruses and HCoV-229E, we tested historical isolates and diagnostic specimens sampled globally over 10 years. Bat viruses were 5- and 6-fold more diversified than HCoV-229E in the RNA-dependent RNA polymerase (RdRp) and spike genes. In phylogenetic analyses, HCoV-229E strains were monophyletic and not intermixed with animal viruses. Bat viruses formed three large clades in close and more distant sister relationships. A recently described 229E-related alpaca virus occupied an intermediate phylogenetic position between bat and human viruses. According to taxonomic criteria, human, alpaca, and bat viruses form a single CoV species showing evidence for multiple recombination events. HCoV-229E and the alpaca virus showed a major deletion in the spike S1 region compared to all bat viruses. Analyses of four full genomes from 229E-related bat CoVs revealed an eighth open reading frame (ORF8) located at the genomic 3' end. ORF8 also existed in the 229E-related alpaca virus. Reanalysis of HCoV-229E sequences showed a conserved transcription regulatory sequence preceding remnants of this ORF, suggesting its loss after acquisition of a 229E-related CoV by humans. These data suggested an evolutionary origin of 229E-related CoVs in hipposiderid bats, hypothetically with camelids as intermediate hosts preceding the establishment of HCoV-229E. IMPORTANCE: The ancestral origins of major human coronaviruses (HCoVs) likely involve bat hosts. Here, we provide conclusive genetic evidence for an evolutionary origin of the common cold virus HCoV-229E in hipposiderid bats by analyzing a large sample of African bats and characterizing several bat viruses on a full-genome level. Our evolutionary analyses show that animal and human viruses are genetically closely related, can exchange genetic material, and form a single viral species. We show that the putative host switches leading to the formation of HCoV-229E were accompanied by major genomic changes, including deletions in the viral spike glycoprotein gene and loss of an open reading frame. We reanalyze a previously described genetically related alpaca virus and discuss the role of camelids as potential intermediate hosts between bat and human viruses. The evolutionary history of HCoV-229E likely shares important characteristics with that of the recently emerged highly pathogenic Middle East respiratory syndrome (MERS) coronavirus.
- MeSH
- Bayesova věta MeSH
- biologická evoluce * MeSH
- Chiroptera virologie MeSH
- DNA primery genetika MeSH
- feces virologie MeSH
- fylogeneze * MeSH
- genetická variace * MeSH
- glykoprotein S, koronavirus genetika MeSH
- lamy virologie MeSH
- lidé MeSH
- lidský koronavirus 229E genetika MeSH
- modely genetické MeSH
- molekulární sekvence - údaje MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- RNA-dependentní RNA-polymerasa genetika MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Ghana MeSH
Maligní melanom je jedním z nejzávažnějších kožních nádorů u lidí i zvířat. Je vysoce odolný vůči konvenčním terapiím a i přes velké úsilí při vývoji nových imunoterapií došlo jen k jejich drobným vylepšením. Proto se jako ideální směr dalšího výzkumu jeví možnost vývoje imunoterapií, které by byly schopné navodit kompletní regresi nádoru, což je ideální výsledek při léčbě jakéhokoliv nádor. Bohužel podmínky potřebné k dosažení úplné regrese nejsou dosud dobře známé. V laboratoři biologie nádorů v Ústavu živočišné fyziologie a genetiky AV ČR je k dispozici model melanomu u miniaturních prasat, u kterého dochází k spontánní regresi nádorů po období progrese spojené s metastázami především do sleziny, plic a lymfatických uzlin, kdy dojde k vyléčení většiny zvířat a pouze asi 5 % umírá na progresi nádoru nebo přidružené komplikace. Další výzkum MeLiM modelu umožní získat nové poznatky o spontánní regresi melanomu a nabídne nové možnosti pro tvorbu účinnějších imunoterapií.
Malignant melanoma is one of the most serious skin cancer diseases in humans and animals. It is highly resistant to conventional therapies and despite major efforts in development of novel immunotherapies there have been only minor improvements. Therefore, an ideal further research should be led towards developing immunotherapies that would be capable of inducing a complete tumor regression. This is the ideal result of the treatment of all tumor. Unfortunately, the conditions required to achieve complete regressions are not well known yet. In the laboratory of tumor biology at the Institute of Animal Physiology and Genetics AS CR an animal model of melanoma is being researched, ie melanoma bearing Libechov minipigs, in which spontaneous tumor regression occurs after a period of progression associated with organ metastases - mainly in the spleen, lungs and lymph nodes. Most animals are completelycured, but around 10 % die of tumor progression or associated complications. Further research of MeLiM model will provide new knowledge about the spontaneous regression of melanoma and offer new possibilities for creating more effective immunotherapy.
- MeSH
- Cyprinodontiformes MeSH
- lidé MeSH
- melanom * MeSH
- miniaturní prasata * genetika MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- spontánní regrese nádoru MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
