engineered cell microenvironments
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T-cell engagers represent a transformative approach to cancer immunotherapy leveraging bispecific and multispecific antibody constructs to redirect T-cell cytotoxicity toward malignant cells. These molecules bridge T cells and tumor cells by simultaneously binding CD3 on T cells and tumor-associated antigens on cancer cells, thereby enabling precise immune targeting even in immunologically "cold" tumors. Recent advancements include conditional T-cell engagers activated by tumor microenvironment proteases to minimize off-tumor toxicity as well as T-cell receptor-based engagers targeting intracellular antigens via MHC presentation. Clinical successes, such as Kimmtrak in metastatic uveal melanoma, underscore good potential of these modalities, while challenges persist in the management of cytokine release syndrome, neurotoxicity, and tumor resistance. Emerging multispecific engagers are aimed at enhancing efficacy via incorporation of costimulatory signals, thus offering a promising trajectory for next-generation immunotherapies. T-cell engagers are also gaining attention in the treatment of autoimmune disorders, where they can be designed to selectively modulate pathogenic immune responses. By targeting autoreactive T or B cells, T-cell engagers hold promise for restoring immune tolerance in such conditions as HLA-B*27-associated autoimmunity subtypes, multiple sclerosis, rheumatoid arthritis, and type 1 diabetes mellitus. Engineering strategies that incorporate inhibitory receptors or tissue-specific antigens may further refine T-cell engagers' therapeutic potential in autoimmunity, by minimizing systemic immunosuppression while preserving immune homeostasis.
- MeSH
- imunoterapie * metody MeSH
- lidé MeSH
- nádorové mikroprostředí imunologie MeSH
- nádory * imunologie terapie MeSH
- protilátky bispecifické terapeutické užití imunologie MeSH
- receptory antigenů T-buněk imunologie metabolismus MeSH
- T-lymfocyty * imunologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Patient-derived organoids (PDOs) and xenografts (PDXs) are powerful tools for personalized medicine in pancreatic cancer (PC) research. This study explores the complementary strengths of PDOs and PDXs in terms of practicality, genetic fidelity, cost, and labor considerations. Among other models like 2D cell cultures, spheroids, cancer-on-chip systems, cell line-derived xenografts (CDX), and genetically engineered mouse models (GEMMs), PDOs and PDXs uniquely balance genetic fidelity and personalized medicine potential, offering distinct advantages over the simplicity of 2D cultures and the advanced, but often resource-intensive, GEMMs and cancer-on-chip systems. PDOs excel in high-throughput drug screening due to their ease of use, lower cost, and shorter experimental timelines. However, they lack a complete tumor microenvironment. Conversely, PDXs offer a more complex microenvironment that closely reflects patient tumors, potentially leading to more clinically relevant results. Despite limitations in size, number of specimens, and engraftment success, PDXs demonstrate significant concordance with patient responses to treatment, highlighting their value in personalized medicine. Both models exhibit significant genetic fidelity, making them suitable for drug sensitivity testing. The choice between PDOs and PDXs depends on the research focus, resource availability, and desired level of microenvironment complexity. PDOs are advantageous for high-throughput screening of a diverse array of potential therapeutic agents due to their relative ease of culture and scalability. PDXs, on the other hand, offer a more physiologically relevant model, allowing for a comprehensive evaluation of drug efficacy and mechanisms of action.
- MeSH
- individualizovaná medicína * metody MeSH
- lidé MeSH
- myši MeSH
- nádorové mikroprostředí účinky léků MeSH
- nádory slinivky břišní * farmakoterapie patologie genetika MeSH
- organoidy * účinky léků patologie MeSH
- protinádorové látky farmakologie terapeutické užití MeSH
- screeningové testy protinádorových léčiv metody MeSH
- xenogenní modely - testy protinádorové aktivity * metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
... and tissue engineering (V. ... ... Rotrekl) 174 -- 12.1 Stem cells - general introduction 174 -- 12.1.1 Stem cells: the beginning 175 -- ... ... Morphogens in the control of stem cell fate 182 -- 12.2.3 Synthetic matrices and artificial microenvironments ... ... 182 -- 12.2.4 Intercellular interactions in the control of stem cell fate 183 -- 12.3 Stem cells in ... ... medicine 184 -- 12.3.1 Mesenchymal stem cells in medicine 184 -- 12.3.2 Pluripotent stem cells in medicine ...
First edition 227 stran : ilustrace ; 30 cm
- Konspekt
- Lékařské vědy. Lékařství
- Učební osnovy. Vyučovací předměty. Učebnice
- NLK Obory
- biologie
- NLK Publikační typ
- učebnice vysokých škol
Východiska: Karcinom prsu je v gynekologických onemocněních celosvětově považován za velkou klinickou výzvu. Exozomy jsou malé vezikuly vzniklé z multicelulárních útvarů, které jsou uvolňovány mnoha buňkami do extracelulárního prostředí a tím se podílí na intercelulární komunikaci prostřednictvím přenosu genetické informace, např. prostřednictvím kódovaných a nekódovaných RNA k cílovým buňkám. Exozomy vytvořené v tumoru jsou považovány za bohatý zdroj microRNA (miRNA), které regulují funkci jiných nádorových buněk v mikroprostředí tumoru. Nicméně přesné mechanizmy, prostřednictvím kterých exozomy odvozené od nádorových buněk ovlivňují sousední buňky, a biologická funkce exozomálních miRNA v receptorových buňkách nejsou ještě dobře objasněny. Materiál a metody: V této studii byly po overexpresi miR-205 v buňkách karcinomu prsu (třída MDA-MB-231) úspěšně izolovány exozomy odvozené od buněk a byly charakterizovány elektronovou mikroskopií a metodou dynamického rozptylu světla. Výsledky: Stanovení míry exprese miR-205 v exozomech uvolňovaných z geneticky upravených buněk potvrdily vysokou expresi této miRNA v exozomech. Bylo také zjištěno, že úprava nádorových exozomů, které nesou tuto miRNA, měla v buňkách karcinomu prsu efekt indukce apoptózy a také měla významný účinek na snížení exprese transkriptu genu Bcl-2 v závislosti na čase (p < 0,001). Závěr: Tato studie naznačuje, že přenos nádorových supresorových miRNA pomocí exozomů by mohl být vhodnou platformou pro přenos nukleových kyselin do těchto buněk a při léčbě karcinomu prsu by mohl být vysoce účinný.
Background: Breast cancer is recognized as a major clinical challenge in gynecological diseases worldwide. Exosomes are small vesicles derived from multicellular bodies that are secreted by many cells into the extracellular environment and thus participate in intercellular communication through the transfer of genetic information such as encoded and non-encoded RNAs to target cells. Tumor-derived exosomes are thought to be a rich source of microRNAs (miRNAs) that can regulate the function of other cancer cells in the tumor microenvironment. However, the exact mechanisms by which tumor cell-derived exosomes affect their neighboring cells, as well as the biological function of exosomal miRNAs in receptor cells, are not well understood. Materials and methods: In this study, after overexpression of miR-205 in breast cancer cells (MDA-MB-231 class), cell-derived exosomes were successfully isolated and characterized by electron microscopy and dynamic light scattering. Results: Determination of miR-205 expression levels in exosomes secreted from engineered cells confirmed the high expression of this miRNA in exosomes. It was also found that treatment of tumor exosomes carrying this miRNA had an apoptotic induction effect and also had a significant effect on reducing the expression of Bcl-2 gene transcript in a time-dependent manner in breast cancer cells (P < 0.001). Conclusion: Overall, this study suggests that exosomal transfer of tumor suppressor miRNAs to cancer cells could be a suitable platform for nucleic acid transfer to these cells and be highly effective in cancer treatment.
- MeSH
- apoptóza účinky léků MeSH
- elektronová mikroskopie metody MeSH
- exozom * genetika izolace a purifikace účinky léků MeSH
- genetická terapie klasifikace metody MeSH
- lidé MeSH
- mikro RNA genetika izolace a purifikace klasifikace účinky léků MeSH
- nádorové buňky kultivované účinky léků MeSH
- nádory prsu * farmakoterapie genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- klinická studie MeSH
The unique properties of stem cells to self-renew and differentiate hold great promise in disease modelling and regenerative medicine. However, more information about basic stem cell biology and thorough characterization of available stem cell lines is needed. This is especially essential to ensure safety before any possible clinical use of stem cells or partially committed cell lines. As proteins are the key effector molecules in the cell, the proteomic characterization of cell lines, cell compartments or cell secretome and microenvironment is highly beneficial to answer above mentioned questions. Nowadays, method of choice for large-scale discovery-based proteomic analysis is mass spectrometry (MS) with data-independent acquisition (DIA). DIA is a robust, highly reproducible, high-throughput quantitative MS approach that enables relative quantification of thousands of proteins in one sample. In the current protocol, we describe a specific variant of DIA known as SWATH-MS for characterization of neural stem cell differentiation. The protocol covers the whole process from cell culture, sample preparation for MS analysis, the SWATH-MS data acquisition on TTOF 5600, the complete SWATH-MS data processing and quality control using Skyline software and the basic statistical analysis in R and MSstats package. The protocol for SWATH-MS data acquisition and analysis can be easily adapted to other samples amenable to MS-based proteomics.
The 2022 Annual Review Issue of The Journal of Pathology, Recent Advances in Pathology, contains 15 invited reviews on research areas of growing importance in pathology. This year, the articles include those that focus on digital pathology, employing modern imaging techniques and software to enable improved diagnostic and research applications to study human diseases. This subject area includes the ability to identify specific genetic alterations through the morphological changes they induce, as well as integrating digital and computational pathology with 'omics technologies. Other reviews in this issue include an updated evaluation of mutational patterns (mutation signatures) in cancer, the applications of lineage tracing in human tissues, and single cell sequencing technologies to uncover tumour evolution and tumour heterogeneity. The tissue microenvironment is covered in reviews specifically dealing with proteolytic control of epidermal differentiation, cancer-associated fibroblasts, field cancerisation, and host factors that determine tumour immunity. All of the reviews contained in this issue are the work of invited experts selected to discuss the considerable recent progress in their respective fields and are freely available online (https://onlinelibrary.wiley.com/journal/10969896). © 2022 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Bone tissue engineering strategy involves the 3D scaffolds and appropriate cell types promoting the replacement of the damaged area. In this work, we aimed to develop a fast and reliable clinically relevant protocol for engineering viable bone grafts, using cryopreserved adipose tissue-derived mesenchymal stromal cells (MSCs) and composite 3D collagen-nano-hydroxyapatite (nanoHA) scaffolds. Xeno- and DMSO-free cryopreserved MSCs were perfusion-seeded into the biomimetic collagen/nanoHA scaffolds manufactured by cryotropic gelation and their osteoregenerative potential was assessed in vitro and in vivo. Cryopreserved MSCs retained the ability to homogenously repopulate the whole volume of the scaffolds during 7 days of post-thaw culture. Moreover, the scaffold provided a suitable microenvironment for induced osteogenic differentiation of cells, confirmed by alkaline phosphatase activity and mineralization. Implantation of collagen-nanoHA cryogels with cryopreserved MSCs accelerated woven bone tissue formation, maturation of bone trabeculae, and vascularization of femur defects in immunosuppressed rats compared to cell-free collagen-nanoHA scaffolds. The established combination of xeno-free cell culture and cryopreservation techniques together with an appropriate scaffold design and cell repopulation approach accelerated the generation of viable bone grafts.
- MeSH
- buněčná diferenciace MeSH
- kolagen farmakologie MeSH
- kryogely * MeSH
- kryoprezervace MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- mezenchymální kmenové buňky * metabolismus MeSH
- osteogeneze MeSH
- proliferace buněk MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Periodontal disease begins as an inflammatory response to a bacterial biofilm deposited around the teeth, which over time leads to the destruction of tooth-supporting structures and consequently tooth loss. Conventional treatment strategies show limited efficacy in promoting regeneration of damaged periodontal tissues. Here, a delivery platform is developed for small extracellular vesicles (sEVs) derived from gingival mesenchymal stem cells (GMSCs) to treat periodontitis. EVs can achieve comparable therapeutic effects to their cells of origin. However, the short half-lives of EVs after their administration along with their rapid diffusion away from the delivery site necessitate frequent administration to achieve therapeutic benefits. To address these issues, "dual delivery" microparticles are engineered enabling microenvironment-sensitive release of EVs by metalloproteinases at the affected site along with antibiotics to suppress bacterial biofilm growth. GMSC sEVs are able to decrease the secretion of pro-inflammatory cytokines by monocytes/macrophages and T cells, suppress T-cell activation, and induce the formation of T regulatory cells (Tregs) in vitro and in a rat model of periodontal disease. One-time administration of immunomodulatory GMSC sEV-decorated microparticles leads to a significant improvement in regeneration of the damaged periodontal tissue. This approach will have potential clinical applications in the regeneration of a variety of tissues.
Natural killer (NK) cells are a family of lymphocytes with a natural ability to kill infected, harmed, or malignantly transformed cells. As these cells are part of the innate immunity, the cytotoxic mechanisms are activated upon recognizing specific patterns without prior antigen sensitization. This recognition is crucial for NK cell function in the maintenance of homeostasis and immunosurveillance. NK cells not only act directly toward malignant cells but also participate in the complex immune response by producing cytokines or cross-talk with other immune cells. Cancer may be seen as a break of all immune defenses when malignant cells escape the immunity and invade surrounding tissues creating a microenvironment supporting tumor progression. This process may be reverted by intervening immune response with immunotherapy, which may restore immune recognition. NK cells are important effector cells for immunotherapy. They may be used for adoptive cell transfer, genetically modified with chimeric antigen receptors, or triggered with appropriate antibodies and other antibody-fragment-based recombinant therapeutic proteins tailored specifically for NK cell engagement. NK cell receptors, responsible for target recognition and activation of cytotoxic response, could also be targeted in immunotherapy, for example, by various bi-, tri-, or multi-specific fusion proteins designed to bridge the gap between tumor markers present on target cells and activation receptors expressed on NK cells. However, this kind of immunoactive therapeutics may be developed only with a deep functional and structural knowledge of NK cell receptor: ligand interactions. This review describes the recent developments in the fascinating protein-engineering field of NK cell immunotherapeutics.
- MeSH
- buňky NK patologie MeSH
- chimerické antigenní receptory * terapeutické užití MeSH
- imunologické faktory MeSH
- imunoterapie adoptivní MeSH
- imunoterapie MeSH
- lidé MeSH
- nádorové mikroprostředí MeSH
- nádory * MeSH
- protinádorové látky * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Východiska: Karcinom prsu je celosvětově považován za hlavní klinický problém u gynekologických onemocnění. Exozomy jsou malé váčky, které vznikají v mnohobuněčných organizmech. Jsou vylučovány různými buňkami do extracelulárního prostředí, a tak se účastní mezibuněčné komunikace tím, že přenáší genetické informace prostřednictvím kódujících a nekódujících RNA do cílových buněk. Exozomy odvozené od nádoru jsou považované za bohatý zdroj mikroRNA (miR) a mohou regulovat funkci nádorových buněk v mikroprostředí nádoru. Přesné mechanismy, kterými exozomy pocházející z nádorových buněk ovlivňují sousední buňky ani biologická funkce exozomálních miR v receptorových buňkách však nejsou dobře známy. Materiál a metody: V této studii byly po nadměrné expresi MiR-205 v buňkách karcinomu prsu (třída MDA-MB-231) úspěšně izolovány buněčné exozomy a charakterizovány pomocí elektronové mikroskopie a dynamického rozptylu světla. Výsledky: Stanovení hladin exprese MiR-205 v exozomech vylučovaných z upravených buněk potvrdilo vysokou expresi této miR. Bylo také zjištěno, že úprava nádorových exozomů nesoucích tuto miR měla indukční účinek na apoptózu a významný účinek na snížení exprese genového transkriptu Bcl-2 v buňkách karcinomu prsu v závislosti na čase (p < 0,001). Závěr: Celkově tato studie naznačuje, že exozomální přenos nádorových supresorových miR do nádorových buněk může být vhodnou platformou pro přenos nukleových kyselin do těchto buněk a může být vysoce účinný při léčbě nádorů.
Background: Breast cancer is recognized as a major clinical challenge in gynecological diseases worldwide. Exosomes are small vesicles derived from multicellular bodies that are secreted by many cells into the extracellular environment and thus participate in intercellular communication through the transfer of genetic information such as encoded and non-encoded RNAs to target cells. Tumor-derived exosomes are thought to be a rich source of microRNAs (miRs) that can regulate the function of other cancer cells in the tumor microenvironment. However, the exact mechanisms by which tumor cell-derived exosomes affect their neighboring cells, as well as the biological function of exosomal miRs in receptor cells, are not well understood. Materials and methods: In this study, after the overexpression of MiR-205 in breast cancer cells (MDA-MB-231 class), cell-derived exosomes were successfully isolated and characterized by electron microscopy and dynamic light scattering. Results: The determination of MiR-205 expression levels in exosomes secreted from engineered cells confirmed the high expression of this miR in exosomes. It was also found that the treatment of tumor exosomes carrying this miR had an apoptotic induction effect and also had a significant effect on reducing the expression of Bcl-2 gene transcript in a time-dependent manner in breast cancer cells (P < 0.001). Conclusion: Overall, this study suggests that exosomal transfer of tumor suppressor miRs to cancer cells could be a suitable platform for nucleic acid transfer to these cells and be highly effective in cancer treatment.
