During innate immune responses, myeloid differentiation primary response 88 (MyD88) functions as a critical signaling adaptor protein integrating stimuli from toll-like receptors (TLR) and the interleukin-1 receptor (IL-1R) family and translates them into specific cellular outcomes. In B cells, somatic mutations in MyD88 trigger oncogenic NF-κB signaling independent of receptor stimulation, which leads to the development of B-cell malignancies. However, the exact molecular mechanisms and downstream signaling targets remain unresolved. We established an inducible system to introduce MyD88 to lymphoma cell lines and performed transcriptomic analysis (RNA-seq) to identify genes differentially expressed by MyD88 bearing the L265P oncogenic mutation. We show that MyD88L265P activates NF-κB signaling and upregulates genes that might contribute to lymphomagenesis, including CD44, LGALS3 (coding Galectin-3), NFKBIZ (coding IkBƺ), and BATF. Moreover, we demonstrate that CD44 can serve as a marker of the activated B-cell (ABC) subtype of diffuse large B-cell lymphoma (DLBCL) and that CD44 expression is correlated with overall survival in DLBCL patients. Our results shed new light on the downstream outcomes of MyD88L265P oncogenic signaling that might be involved in cellular transformation and provide novel therapeutical targets.

• MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• antigeny CD44 genetika   metabolismus   MeSH
• difúzní velkobuněčný B-lymfom * patologie   MeSH
• galektin 3 metabolismus   MeSH
• lidé MeSH
• mutace MeSH
• myeloidní diferenciační faktor 88 genetika   metabolismus   MeSH
• NF-kappa B * genetika   metabolismus   MeSH
• stanovení celkové genové exprese MeSH
• transkripční faktory bZIP genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Galectin-3 (Gal-3) participates in many cancer-related metabolic processes. The inhibition of overexpressed Gal-3 by, e.g., β-galactoside-derived inhibitors is hence promising for cancer treatment. The multivalent presentation of such inhibitors on a suitable biocompatible carrier can enhance the overall affinity to Gal-3 and favorably modify the interaction with Gal-3-overexpressing cells. We synthesized a library of C-3 aryl-substituted thiodigalactoside inhibitors and their multivalent N-(2-hydroxypropyl)methacrylamide (HPMA)-based counterparts with two different glycomimetic contents. Glycopolymers with a higher content of glycomimetic exhibited a higher affinity to Gal-3 as assessed by ELISA and biolayer interferometry. Among them, four candidates (with 4-acetophenyl, 4-cyanophenyl, 4-fluorophenyl, and thiophen-3-yl substitution) were selected for further evaluation in cancer-related experiments in cell cultures. These glycopolymers inhibited Gal-3-induced processes in cancer cells. The cyanophenyl-substituted glycopolymer exhibited the strongest antiproliferative, antimigratory, antiangiogenic, and immunoprotective properties. The prepared glycopolymers appear to be prospective modulators of the tumor microenvironment applicable in the therapy of Gal-3-associated cancers.

• MeSH
• galektin 3 * metabolismus   MeSH
• prospektivní studie MeSH
• thiogalaktosidy * farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Galectin-3 (Gal-3) is a promising target in cancer therapy with a high therapeutic potential due to its abundant localization within the tumor tissue and its involvement in tumor development and proliferation. Potential clinical application of Gal-3-targeted inhibitors is often complicated by their insufficient selectivity or low biocompatibility. Nanomaterials based on N-(2-hydroxypropyl)methacrylamide (HPMA) nanocarrier are attractive for in vivo application due to their good water solubility and lack of toxicity and immunogenicity. Their conjugation with tailored carbohydrate ligands can yield specific glyconanomaterials applicable for targeting biomedicinally relevant lectins like Gal-3. RESULTS: In the present study we describe the synthesis and the structure-affinity relationship study of novel Gal-3-targeted glyconanomaterials, based on hydrophilic HPMA nanocarriers. HPMA nanocarriers decorated with varying amounts of Gal-3 specific epitope GalNAcβ1,4GlcNAc (LacdiNAc) were analyzed in a competitive ELISA-type assay and their binding kinetics was described by surface plasmon resonance. We showed the impact of various linker types and epitope distribution on the binding affinity to Gal-3. The synthesis of specific functionalized LacdiNAc epitopes was accomplished under the catalysis by mutant β-N-acetylhexosaminidases. The glycans were conjugated to statistic HPMA copolymer precursors through diverse linkers in a defined pattern and density using Cu(I)-catalyzed azide-alkyne cycloaddition. The resulting water-soluble and structurally flexible synthetic glyconanomaterials exhibited affinity to Gal-3 in low μM range. CONCLUSIONS: The results of this study reveal the relation between the linker structure, glycan distribution and the affinity of the glycopolymer nanomaterial to Gal-3. They pave the way to specific biomedicinal glyconanomaterials that target Gal-3 as a therapeutic goal in cancerogenesis and other disorders.

• MeSH
• akrylamidy chemie   metabolismus   MeSH
• galektin 3 metabolismus   MeSH
• glykokonjugáty chemie   metabolismus   MeSH
• lékové transportní systémy * MeSH
• lidé MeSH
• nanostruktury chemie   MeSH
• nosiče léků chemie   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Elevated levels of galectin-3 are associated with tumorigenesis. Its inhibition with high-affinity carbohydrate ligands opens new therapeutic routes. Targeting of intracellular galectin-3 is challenging for polar inhibitors like carbohydrates. We demonstrate the potential of novel biomedical research tools, glycocalix[4]arenes, to enter epithelial cells, which may allow their interaction with galectin-3.

• MeSH
• buněčná membrána MeSH
• galektin 3 * MeSH
• galektiny MeSH
• glykokalyx * MeSH
• sacharidy farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• buněčná diferenciace MeSH
• finanční podpora výzkumu jako téma MeSH
• galektin 1 metabolismus   MeSH
• galektin 3 metabolismus   MeSH
• keratinocyty metabolismus   MeSH
• keratiny metabolismus   MeSH
• kmenové buňky metabolismus   MeSH
• lidé MeSH
• stárnutí metabolismus   MeSH
• Check Tag
• lidé MeSH

The development of efficient galectin-3 (Gal-3) inhibitors draws attention in the field of anti-cancer therapy, especially due to the prominent role of extra- and intracellular Gal-3 in vital processes of cancerogenesis, such as immunosuppression, stimulation of tumor cells proliferation, survival, invasion, apoptotic resistance, and metastasis formation and progression. Here, by combining poly-LacNAc (Galβ4GlcNAc)-derived oligosaccharides with N-(2-hydroxypropyl) methacrylamide (HPMA) copolymers, we synthesized multivalent glycopolymer inhibitors with a high potential to target extracellular and intracellular Gal-3. The inhibitory capabilities of the best conjugate in the studied series were in the nanomolar range proving the excellent Gal-3 inhibitory potential. Moreover, thorough investigation of the inhibitory effect in the biological conditions showed that the glycopolymers strongly inhibited Gal-3-induced apoptosis of T lymphocytes and suppressed migration and spreading of colorectal, breast, melanoma, and prostate cancer cells. In sum, the strong inhibitory activity toward Gal-3, combined with favorable pharmacokinetics of HPMA copolymers ensuring enhanced tumor accumulation via the enhanced permeability and retention effect, nominate the glycopolymers containing LacdiNAc-LacNAc (GalNAcβ4GlcNAcβ3Galβ4GlcNAc) tetrasaccharide as promising tools for preclinical in anti-cancer therapy evaluation.

• MeSH
• apoptóza * MeSH
• galektin 3 * MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• pohyb buněk MeSH
• polymery MeSH
• T-lymfocyty MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Galectin-3 (Gal-3) is recognized as a prognostic marker in several cancer types. Its involvement in tumor development and proliferation makes this lectin a promising target for early cancer diagnosis and anti-cancer therapies. Gal-3 recognizes poly-N-acetyllactosamine (LacNAc)-based carbohydrate motifs of glycoproteins and glycolipids with a high specificity for internal LacNAc epitopes. This study analyzes the mode and kinetics of binding of Gal-3 to a series of multivalent neo-glycoproteins presenting complex poly-LacNAc-based oligosaccharide ligands on a scaffold of bovine serum albumin. These neo-glycoproteins rank among the strongest Gal-3 ligands reported, with Kd reaching sub-nanomolar values as determined by surface plasmon resonance. Significant differences in the binding kinetics were observed within the ligand series, showing the tetrasaccharide capped with N,N'-diacetyllactosamine (LacdiNAc) as the strongest ligand of Gal-3 in this study. A molecular model of the Gal-3 carbohydrate recognition domain with docked oligosaccharide ligands is presented that shows the relations in the binding site at the molecular level. The neo-glycoproteins presented herein may be applied for selective recognition of Gal-3 both on the cell surface and in blood serum.

• MeSH
• galektin 3 chemie   metabolismus   MeSH
• glykoproteiny chemie   farmakologie   MeSH
• laktosa analogy a deriváty   chemie   MeSH
• lidé MeSH
• ligandy MeSH
• sérový albumin hovězí chemie   MeSH
• simulace molekulového dockingu * MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Galectin-3 is an immunomodulatory protein with binding capacity for various glycoconjugates including IgE. It has been shown to be produced by epidermal keratinocytes and is present on the surfaces of skin Langerhans cells (LC). Therefore, it may have a role in the pathogenesis of various skin diseases, such as atopic dermatitis. To study the expression of galectin-3 in LC, we used, in addition to specific antibodies, a panel of synthetic, carrier-immobilized, specific oligosaccharides of the A- and B-histo-blood group, which are recognized by this lectin. In the mean time, Birbeck granules were visualized with an anti-Lag antibody. The double labeling experiments showed a remarkable colocalization of signals for Lag antigen (Birbeck granules) and galectin-3, as well as the binding sites for A- and B-histo-blood group trisaccharides. The specificity of the oligosaccharide binding was demonstrated by the lack of binding by Le(c), Le(d) (H blood group antigen), and sLe(x), which are not recognized by galectin-3. These results suggest that galectin-3 is present in Birbeck granules, where it retains reactivity for its glycoligands.

• MeSH
• ABO systém krevních skupin * metabolismus   MeSH
• cytoplazmatická granula imunologie   MeSH
• diferenciační antigeny genetika   metabolismus   MeSH
• galektin 3 MeSH
• Langerhansovy buňky imunologie   metabolismus   MeSH
• lidé MeSH
• protilátky imunologie   MeSH
• trisacharidy * metabolismus   MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

• Klíčová slova
• reparace, glykobiologie, mezibuněčné interakce,
• MeSH
• fibroblasty cytologie   MeSH
• galektin 1 * aplikace a dávkování   fyziologie   sekrece   MeSH
• galektin 3 * aplikace a dávkování   fyziologie   sekrece   MeSH
• hojení ran * fyziologie   MeSH
• kůže růst a vývoj   zranění   MeSH
• lidé MeSH
• mezibuněčné spoje MeSH
• myši zranění   MeSH
• nádorová transformace buněk MeSH
• prospektivní studie MeSH
• regenerace MeSH
• techniky in vitro MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši zranění   MeSH
• zvířata MeSH
• Publikační typ
• grafy a diagramy MeSH

Cílem studie bylo pomocí nepřímé imunohistochemie zjistit expresi galektinu-3 (Gal3), cytokeratinu 19 (CK19), neural cell adhesion molecule (NCAM) a E-cadherinu (Ecad) ve variantách papilárního karcinomu štítné žlázy (PTC) s ohledem na možné využití těchto markerů v bioptické diagnostice. Soubor tvořilo 84 případů – 36 klasických variant PTC (cPTC), 26 folikulárních variant PTC (fPTC) a 22 papilárních mikrokarcinomů (mPTC). Gal3 byl exprimován ve 36/36 (100 %) cPTC, ve 24/26 (92 %) fPTC a v 19/22 (86 %) mPTC. Exprese CK19 byla zastižena ve 34/36 (94 %) cPTC, v 17/26 (65 %) fPTC a ve 13/22 (59 %) mPTC. Exprese NCAM byla prokázána v 5/36 (14 %) cPTC, v 7/26 (27 %) fPTC a v 9/22 (41 %) mPTC. Ecad byl exprimován ve 23/36 (64 %) cPTC, v 17/26 (65 %) fPTC a v 18/22 (82 %) mPTC. V expresi CK19 byl zjištěn signifikantní rozdíl mezi cPTC versus fPTC a mPTC (p < 0,001). Dále byla prokázána signifikantní korelace mezi expresí CK19 a ztrátou exprese Ecad ve vztahu k šíření nádoru mimo štítnou žlázu (p = 0,001, p = 0,04). Detekci exprese Gal3 a CK19 lze tedy doporučit jako pomocné kritérium v diagnostice PTC, nicméně je třeba upozornit na nižší expresi CK19 ve fPTC a v mPTC. K detailnímu posouzení úlohy CK19 a Ecad při šíření PTC mimo štítnou žlázu je třeba dalších studií.

The immunohistochemical expression of galectin-3 (Gal3), cytokeratin 19 (CK19), neural cell adhesion molecule (NCAM), and E-cadherin (Ecad) was evaluated to assess their use in diagnostics of papillary thyroid carcinoma (PTC). A total of 84 PTCs - 36 classical variants (cPTCs), 26 follicular variants (fPTCs), and 22 papillary microcarcinomas (mPTCs) were studied. Expression of Gal3 was found in 36/36 (100%) cPTCs, 24/26 (92%) fPTCs, and 19/22 (86%) mPTCs. CK19 expression was detected in 34/36 (94%) cPTCs, 17/26 (65%) fPTCs, and 13/22 (59%) mPTCs. Expression of NCAM was seen in 5/36 (14%) cPTCs, 7/26 (27%) fPTCs, and 9/22 (41%) mPTCs. Ecad expression was found in 23/36 (64%) cPTCs, 17/26 (65%) fPTCs, and 18/22 (82%) mPTCs. A significant difference in CK19 expression was observed between cPTC and both fPTC and mPTC (p < 0.001). Furthermore, extrathyroid tumor spread significantly correlated with both level of CK19 expression and loss of Ecad expression (p = 0.001, p = 0.04). Our findings suggest that Gal3 and CK19 are useful markers for PTC, although decreased CK19 expression in mPTC and fPTC must be considered. Furthermore, CK19 and Ecad may play a role in extrathyroid tumor spread.

• MeSH
• diferenciální diagnóza MeSH
• finanční podpora výzkumu jako téma MeSH
• galektin 3 diagnostické užití   MeSH
• kadheriny diagnostické užití   MeSH
• keratin-19 diagnostické užití   MeSH
• molekuly buněčné adheze nervové diagnostické užití   MeSH
• nádory štítné žlázy klasifikace   patologie   MeSH
• papilární karcinom klasifikace   patologie   MeSH
• Publikační typ
• hodnotící studie MeSH