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.

• Klíčová slova
• CD44, DLBCL, Galectin-3, LGALS3, MYD88, NF-kB, lymphoma, oncogenic signaling,
• 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
• NF-kappa B * genetika   metabolismus   MeSH
• protein MyD88 genetika   metabolismus   MeSH
• stanovení celkové genové exprese MeSH
• transkripční faktory bZIP genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• antigeny CD44 MeSH
• BATF protein, human MeSH Prohlížeč
• CD44 protein, human MeSH Prohlížeč
• galektin 3 MeSH
• NF-kappa B * MeSH
• NFKBIZ protein, human MeSH Prohlížeč
• protein MyD88 MeSH
• transkripční faktory bZIP MeSH

Acute lung injury (ALI) caused by lipopolysaccharide (LPS) is a common, severe clinical syndrome. Injury caused by inflammation and oxidative stress in vascular endothelial and alveolar epithelial cells is a vital process in the pathogenesis of ALI. Toll-like receptor 9 (TLR9) is highly expressed in LPS-induced ALI rats. In this study, Beas-2B human pulmonary epithelial cells and A549 alveolar epithelial cells were stimulated by LPS, resulting in the upregulation of TLR9 in a concentrationdependent manner. Furthermore, TLR9 overexpression and interference vectors were transfected before LPS administration to explore the role of TLR9 in LPS-induced ALI in vitro. The findings revealed that inhibition of TLR9 reduced inflammation and oxidative stress while suppressing apoptosis of LPS-induced Beas-2B and A549 cells, whereas TLR9 overexpression aggravated these conditions. Moreover, TLR9 inhibition resulted in downregulated protein expression of myeloid differentiation protein 88 (MyD88) and activator activator protein 1 (AP-1), as well as phosphorylation of nuclear factor-?B (NF-kappaB), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK). The phosphorylation of extracellular-regulated protein kinases 1/2 was upregulated compared to that of cells subjected to only LPS administration, and this was reversed by TLR9 overexpression. These results indicate that inhibition of TLR9 plays a protective role against LPS-induced inflammation and oxidative stress in Beas-2B and A549 cells, possibly via the MyD88/NF-kappaB and MyD88/MAPKs/AP-1 pathways.

• MeSH
• akutní poškození plic * chemicky indukované   metabolismus   prevence a kontrola   MeSH
• epitelové buňky patologie   MeSH
• krysa rodu Rattus MeSH
• lipopolysacharidy * metabolismus   toxicita   MeSH
• NF-kappa B metabolismus   MeSH
• oxidační stres MeSH
• protein MyD88 metabolismus   MeSH
• signální transdukce MeSH
• toll-like receptor 4 metabolismus   MeSH
• toll-like receptor 9 genetika   metabolismus   MeSH
• transkripční faktor AP-1 metabolismus   MeSH
• zánět chemicky indukované   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• lipopolysacharidy * MeSH
• Myd88 protein, rat MeSH Prohlížeč
• NF-kappa B MeSH
• protein MyD88 MeSH
• Tlr9 protein, rat MeSH Prohlížeč
• toll-like receptor 4 MeSH
• toll-like receptor 9 MeSH
• transkripční faktor AP-1 MeSH

Next-generation sequencing has revealed novel recurrent mutations in chronic lymphocytic leukemia, particularly in patients with aggressive disease. Here, we explored targeted re-sequencing as a novel strategy to assess the mutation status of genes with prognostic potential. To this end, we utilized HaloPlex targeted enrichment technology and designed a panel including nine genes: ATM, BIRC3, MYD88, NOTCH1, SF3B1 and TP53, which have been linked to the prognosis of chronic lymphocytic leukemia, and KLHL6, POT1 and XPO1, which are less characterized but were found to be recurrently mutated in various sequencing studies. A total of 188 chronic lymphocytic leukemia patients with poor prognostic features (unmutated IGHV, n=137; IGHV3-21 subset #2, n=51) were sequenced on the HiSeq 2000 and data were analyzed using well-established bioinformatics tools. Using a conservative cutoff of 10% for the mutant allele, we found that 114/180 (63%) patients carried at least one mutation, with mutations in ATM, BIRC3, NOTCH1, SF3B1 and TP53 accounting for 149/177 (84%) of all mutations. We selected 155 mutations for Sanger validation (variant allele frequency, 10-99%) and 93% (144/155) of mutations were confirmed; notably, all 11 discordant variants had a variant allele frequency between 11-27%, hence at the detection limit of conventional Sanger sequencing. Technical precision was assessed by repeating the entire HaloPlex procedure for 63 patients; concordance was found for 77/82 (94%) mutations. In summary, this study demonstrates that targeted next-generation sequencing is an accurate and reproducible technique potentially suitable for routine screening, eventually as a stand-alone test without the need for confirmation by Sanger sequencing.

• MeSH
• alely MeSH
• chronická lymfatická leukemie diagnóza   genetika   metabolismus   patologie   MeSH
• exprese genu MeSH
• fosfoproteiny genetika   metabolismus   MeSH
• frekvence genu MeSH
• lidé MeSH
• malý jaderný ribonukleoprotein U2 genetika   metabolismus   MeSH
• mutace * MeSH
• nádorové proteiny genetika   metabolismus   MeSH
• nádorový supresorový protein p53 genetika   metabolismus   MeSH
• prognóza MeSH
• protein MyD88 genetika   metabolismus   MeSH
• receptor Notch1 genetika   metabolismus   MeSH
• sestřihové faktory MeSH
• vysoce účinné nukleotidové sekvenování * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fosfoproteiny MeSH
• malý jaderný ribonukleoprotein U2 MeSH
• MYD88 protein, human MeSH Prohlížeč
• nádorové proteiny MeSH
• nádorový supresorový protein p53 MeSH
• NOTCH1 protein, human MeSH Prohlížeč
• protein MyD88 MeSH
• receptor Notch1 MeSH
• sestřihové faktory MeSH
• SF3B1 protein, human MeSH Prohlížeč
• TP53 protein, human MeSH Prohlížeč

Mammalian TLRs in adult animals serve indispensable functions in establishing innate and adaptive immunity and contributing to the homeostasis of surrounding tissues. However, the expression and function of TLRs during mammalian embryonic development has not been studied so far. Here, we show that CD45(+) CD11b(+) F4/80(+) macrophages from 10.5-day embryo (E10.5) co-express TLRs and CD14. These macrophages, which have the capability to engulf both apoptotic cells and bacteria, secrete a broad spectrum of proinflammatory cytokines and chemokines upon TLR stimulation, demonstrating that their TLRs are functional. Comparative microarray analysis revealed an additional set of genes that were significantly upregulated in E10.5 TLR2(+) CD11b(+) macrophages. This analysis, together with our genetic, microscopic, and biochemical evidence, showed that embryonic phagocytes express protein machinery that is essential for the recycling of cellular iron and that this expression can be regulated by TLR engagement in a MyD88-dependent manner, leading to typical inflammatory M1 responses. These results characterize the utility of TLRs as suitable markers for early embryonic phagocytes as well as molecular triggers of cellular responses, the latter being demonstrated by the involvement of TLRs in an inflammation-mediated regulation of embryonic homeostasis via iron metabolism.

• Klíčová slova
• Embryonic macrophages, Ferroportin, Gene expression microarray, Iron metabolism, TLR stimulation,
• MeSH
• diferenciační antigeny genetika   imunologie   metabolismus   MeSH
• embryo savčí cytologie   imunologie   metabolismus   MeSH
• makrofágy cytologie   imunologie   metabolismus   MeSH
• myši knockoutované MeSH
• myši MeSH
• protein MyD88 genetika   imunologie   metabolismus   MeSH
• signální transdukce fyziologie   MeSH
• toll-like receptor 2 genetika   imunologie   metabolismus   MeSH
• vývojová regulace genové exprese genetika   imunologie   MeSH
• zánět genetika   imunologie   MeSH
• železo imunologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• diferenciační antigeny MeSH
• Myd88 protein, mouse MeSH Prohlížeč
• protein MyD88 MeSH
• Tlr2 protein, mouse MeSH Prohlížeč
• toll-like receptor 2 MeSH
• železo MeSH