Identification and characterization of stromal-like cells with CD207+/low CD1a+/low phenotype derived from histiocytic lesions – a perspective in vitro model for drug testing
Abstract Background Histiocytoses are rare disorders manifested by increased proliferation of pathogenic myeloid cells sharing histological features with macrophages or dendritic cells and accumulating in various organs, i.a., bone and skin. Pre-clinical in vitro models that could be used to determi...
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| Format: | Article |
| Language: | English |
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BMC
2024-02-01
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| Series: | BMC Cancer |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12885-023-11807-0 |
| _version_ | 1797274289839276032 |
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| author | Agnieszka Śmieszek Klaudia Marcinkowska Zofia Małas Mateusz Sikora Martyna Kępska Beata A. Nowakowska Marta Deperas Marta Smyk Carlos Rodriguez-Galindo Anna Raciborska |
| author_facet | Agnieszka Śmieszek Klaudia Marcinkowska Zofia Małas Mateusz Sikora Martyna Kępska Beata A. Nowakowska Marta Deperas Marta Smyk Carlos Rodriguez-Galindo Anna Raciborska |
| author_sort | Agnieszka Śmieszek |
| collection | DOAJ |
| description | Abstract Background Histiocytoses are rare disorders manifested by increased proliferation of pathogenic myeloid cells sharing histological features with macrophages or dendritic cells and accumulating in various organs, i.a., bone and skin. Pre-clinical in vitro models that could be used to determine molecular pathways of the disease are limited, hence research on histiocytoses is challenging. The current study compares cytophysiological features of progenitor, stromal-like cells derived from histiocytic lesions (sl-pHCs) of three pediatric patients with different histiocytoses types and outcomes. The characterized cells may find potential applications in drug testing. Methods Molecular phenotype of the cells, i.e. expression of CD1a and CD207 (langerin), was determined using flow cytometry. Cytogenetic analysis included GTG-banded metaphases and microarray (aCGH) evaluation. Furthermore, the morphology and ultrastructure of cells were evaluated using a confocal and scanning electron microscope. The microphotographs from the confocal imaging were used to reconstruct the mitochondrial network and its morphology. Basic cytophysiological parameters, such as viability, mitochondrial activity, and proliferation, were analyzed using multiple cellular assays, including Annexin V/7-AAD staining, mitopotential analysis, BrdU test, clonogenicity analysis, and distribution of cells within the cell cycle. Biomarkers potentially associated with histiocytoses progression were determined using RT-qPCR at mRNA, miRNA and lncRNA levels. Intracellular accumulation of histiocytosis-specific proteins was detected with Western blot. Cytotoxicyty and IC50 of vemurafenib and trametinib were determined with MTS assay. Results Obtained cellular models, i.e. RAB-1, HAN-1, and CHR-1, are heterogenic in terms of molecular phenotype and morphology. The cells express CD1a/CD207 markers characteristic for dendritic cells, but also show intracellular accumulation of markers characteristic for cells of mesenchymal origin, i.e. vimentin (VIM) and osteopontin (OPN). In subsequent cultures, cells remain viable and metabolically active, and the mitochondrial network is well developed, with some distinctive morphotypes noted in each cell line. Cell-specific transcriptome profile was noted, providing information on potential new biomarkers (non-coding RNAs) with diagnostic and prognostic features. The cells showed different sensitivity to vemurafenib and trametinib. Conclusion Obtained and characterized cellular models of stromal-like cells derived from histiocytic lesions can be used for studies on histiocytosis biology and drug testing. |
| first_indexed | 2024-03-07T14:56:17Z |
| format | Article |
| id | doaj.art-3f63d3dff8ab47c3833118b32a172bd0 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-03-07T14:56:17Z |
| publishDate | 2024-02-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-3f63d3dff8ab47c3833118b32a172bd02024-03-05T19:24:06ZengBMCBMC Cancer1471-24072024-02-0124112310.1186/s12885-023-11807-0Identification and characterization of stromal-like cells with CD207+/low CD1a+/low phenotype derived from histiocytic lesions – a perspective in vitro model for drug testingAgnieszka Śmieszek0Klaudia Marcinkowska1Zofia Małas2Mateusz Sikora3Martyna Kępska4Beata A. Nowakowska5Marta Deperas6Marta Smyk7Carlos Rodriguez-Galindo8Anna Raciborska9Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life SciencesDepartment of Experimental Biology, Faculty of Biology and Animal Science, Wroclaw University of Environmental and Life SciencesDepartment of Oncology and Surgical Oncology for Children and Youth, Institute of Mother and ChildDepartment of Experimental Biology, Faculty of Biology and Animal Science, Wroclaw University of Environmental and Life SciencesDepartment of Experimental Biology, Faculty of Biology and Animal Science, Wroclaw University of Environmental and Life SciencesMedical Genetics Department, Cytogenetics Laboratory, Institute of Mother and ChildMedical Genetics Department, Cytogenetics Laboratory, Institute of Mother and ChildMedical Genetics Department, Cytogenetics Laboratory, Institute of Mother and Child Jude Children’s Research HospitalDepartment of Oncology and Surgical Oncology for Children and Youth, Institute of Mother and ChildAbstract Background Histiocytoses are rare disorders manifested by increased proliferation of pathogenic myeloid cells sharing histological features with macrophages or dendritic cells and accumulating in various organs, i.a., bone and skin. Pre-clinical in vitro models that could be used to determine molecular pathways of the disease are limited, hence research on histiocytoses is challenging. The current study compares cytophysiological features of progenitor, stromal-like cells derived from histiocytic lesions (sl-pHCs) of three pediatric patients with different histiocytoses types and outcomes. The characterized cells may find potential applications in drug testing. Methods Molecular phenotype of the cells, i.e. expression of CD1a and CD207 (langerin), was determined using flow cytometry. Cytogenetic analysis included GTG-banded metaphases and microarray (aCGH) evaluation. Furthermore, the morphology and ultrastructure of cells were evaluated using a confocal and scanning electron microscope. The microphotographs from the confocal imaging were used to reconstruct the mitochondrial network and its morphology. Basic cytophysiological parameters, such as viability, mitochondrial activity, and proliferation, were analyzed using multiple cellular assays, including Annexin V/7-AAD staining, mitopotential analysis, BrdU test, clonogenicity analysis, and distribution of cells within the cell cycle. Biomarkers potentially associated with histiocytoses progression were determined using RT-qPCR at mRNA, miRNA and lncRNA levels. Intracellular accumulation of histiocytosis-specific proteins was detected with Western blot. Cytotoxicyty and IC50 of vemurafenib and trametinib were determined with MTS assay. Results Obtained cellular models, i.e. RAB-1, HAN-1, and CHR-1, are heterogenic in terms of molecular phenotype and morphology. The cells express CD1a/CD207 markers characteristic for dendritic cells, but also show intracellular accumulation of markers characteristic for cells of mesenchymal origin, i.e. vimentin (VIM) and osteopontin (OPN). In subsequent cultures, cells remain viable and metabolically active, and the mitochondrial network is well developed, with some distinctive morphotypes noted in each cell line. Cell-specific transcriptome profile was noted, providing information on potential new biomarkers (non-coding RNAs) with diagnostic and prognostic features. The cells showed different sensitivity to vemurafenib and trametinib. Conclusion Obtained and characterized cellular models of stromal-like cells derived from histiocytic lesions can be used for studies on histiocytosis biology and drug testing.https://doi.org/10.1186/s12885-023-11807-0HistiocytosesLangerhans cell histiocytosisRare disordersCell linesIn vitro studyCellular model |
| spellingShingle | Agnieszka Śmieszek Klaudia Marcinkowska Zofia Małas Mateusz Sikora Martyna Kępska Beata A. Nowakowska Marta Deperas Marta Smyk Carlos Rodriguez-Galindo Anna Raciborska Identification and characterization of stromal-like cells with CD207+/low CD1a+/low phenotype derived from histiocytic lesions – a perspective in vitro model for drug testing BMC Cancer Histiocytoses Langerhans cell histiocytosis Rare disorders Cell lines In vitro study Cellular model |
| title | Identification and characterization of stromal-like cells with CD207+/low CD1a+/low phenotype derived from histiocytic lesions – a perspective in vitro model for drug testing |
| title_full | Identification and characterization of stromal-like cells with CD207+/low CD1a+/low phenotype derived from histiocytic lesions – a perspective in vitro model for drug testing |
| title_fullStr | Identification and characterization of stromal-like cells with CD207+/low CD1a+/low phenotype derived from histiocytic lesions – a perspective in vitro model for drug testing |
| title_full_unstemmed | Identification and characterization of stromal-like cells with CD207+/low CD1a+/low phenotype derived from histiocytic lesions – a perspective in vitro model for drug testing |
| title_short | Identification and characterization of stromal-like cells with CD207+/low CD1a+/low phenotype derived from histiocytic lesions – a perspective in vitro model for drug testing |
| title_sort | identification and characterization of stromal like cells with cd207 low cd1a low phenotype derived from histiocytic lesions a perspective in vitro model for drug testing |
| topic | Histiocytoses Langerhans cell histiocytosis Rare disorders Cell lines In vitro study Cellular model |
| url | https://doi.org/10.1186/s12885-023-11807-0 |
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