AIM: In this study, we analysed the post-translational modification of receptor tyrosine kinase-like orphan receptor (Ror1). Ror1 is highly upregulated in B cells of patients with chronic lymphocytic leukaemia (CLL). Molecularly, Ror1 acts as the Wnt receptor in the non-canonical Wnt pathway. METHODS: The level of Ror1 glycosylation in HEK293 cells and in primary human CLL cells was analysed by treatment of inhibitors interfering with different steps of glycosylation process and by direct treatment of cell lysates with N-glycosidase. Ror1 ubiquitination was determined by ubiquitination assay. Functional consequences of post-translational modifications were analysed by immunohistochemistry and by analysis of cell surface proteins. Differences in Ror1 glycosylation were confirmed by analysis of 14 samples of B cells from CLL patients. RESULTS: We demonstrate that Ror1 is extensively modified by N-linked glycosylation. Glycosylation produces several variants of Ror1 with electrophoretic migration of approx. 100, 115 and 130 kDa. Inhibition of glycosylation interferes with cell surface localization of the 130-kDa variant of Ror1 and prevents Ror1-induced formation of filopodia. Moreover, we show that 130-kDa Ror1 is mono-ubiquitinated. Furthermore, individual CLL patients show striking differences in the electrophoretic migration of Ror1, which correspond to the level of glycosylation. CONCLUSION: Our data show that Ror1 undergoes complex post-translational modifications by glycosylation and mono-ubiquitination. These modifications regulate Ror1 localization and signalling, and are highly variable among individual CLL patients. These may suggest that Ror1 signals only in a subset of CLL patients despite Ror1 levels are ubiquitously high in all CLL patients.

• MeSH
• B-lymfocyty metabolismus   MeSH
• CHO buňky MeSH
• chronická lymfatická leukemie metabolismus   MeSH
• Cricetulus MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• glykosylace MeSH
• HEK293 buňky MeSH
• imunohistochemie MeSH
• konfokální mikroskopie MeSH
• křečci praví MeSH
• lidé MeSH
• molekulová hmotnost MeSH
• posttranslační úpravy proteinů účinky léků   MeSH
• průtoková cytometrie MeSH
• pseudopodia metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• sirotčí receptory podobné receptoru tyrosinkinasy chemie   genetika   metabolismus   MeSH
• transfekce MeSH
• transport proteinů MeSH
• ubikvitinace MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Chronic lymphocytic leukemia is a disease with up-regulated expression of the transmembrane tyrosine-protein kinase ROR1, a member of the Wnt/planar cell polarity pathway. In this study, we identified COBLL1 as a novel interaction partner of ROR1. COBLL1 shows clear bimodal expression with high levels in chronic lymphocytic leukemia patients with mutated IGHV and approximately 30% of chronic lymphocytic leukemia patients with unmutated IGHV. In the remaining 70% of chronic lymphocytic leukemia patients with unmutated IGHV, COBLL1 expression is low. Importantly, chronic lymphocytic leukemia patients with unmutated IGHV and high COBLL1 have an unfavorable disease course with short overall survival and time to second treatment. COBLL1 serves as an independent molecular marker for overall survival in chronic lymphocytic leukemia patients with unmutated IGHV. In addition, chronic lymphocytic leukemia patients with unmutated IGHV and high COBLL1 show impaired motility and chemotaxis towards CCL19 and CXCL12 as well as enhanced B-cell receptor signaling pathway activation demonstrated by increased PLCγ2 and SYK phosphorylation after IgM stimulation. COBLL1 expression also changes during B-cell maturation in non-malignant secondary lymphoid tissue with a higher expression in germinal center B cells than naïve and memory B cells. Our data thus suggest COBLL1 involvement not only in chronic lymphocytic leukemia but also in B-cell development. In summary, we show that expression of COBLL1, encoding novel ROR1-binding partner, defines chronic lymphocytic leukemia subgroups with a distinct response to microenvironmental stimuli, and independently predicts survival of chronic lymphocytic leukemia with unmutated IGHV.

• MeSH
• analýza přežití MeSH
• chronická lymfatická leukemie klasifikace   diagnóza   genetika   mortalita   MeSH
• lidé MeSH
• mutace MeSH
• pohyb buněk MeSH
• polarita buněk MeSH
• prognóza MeSH
• signální dráha Wnt MeSH
• sirotčí receptory podobné receptoru tyrosinkinasy metabolismus   MeSH
• těžké řetězce imunoglobulinů genetika   MeSH
• transkripční faktory metabolismus   MeSH
• variabilní oblast imunoglobulinu genetika   MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• B-lymfocyty cytologie   MeSH
• chronická lymfatická leukemie diagnóza   patologie   MeSH
• imunomagnetická separace metody   MeSH
• lidé MeSH
• metody MeSH
• sirotčí receptory podobné receptoru tyrosinkinasy imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH

PURPOSE: ROR1, a receptor in the noncanonical Wnt/planar cell polarity (PCP) pathway, is upregulated in malignant B cells of chronic lymphocytic leukemia (CLL) patients. It has been shown that the Wnt/PCP pathway drives pathogenesis of CLL, but which factors activate the ROR1 and PCP pathway in CLL cells remains unclear. EXPERIMENTAL DESIGN: B lymphocytes from the peripheral blood of CLL patients were negatively separated using RosetteSep (StemCell) and gradient density centrifugation. Relative expression of WNT5A, WNT5B, and ROR1 was assessed by quantitative real-time PCR. Protein levels, protein interaction, and downstream signaling were analyzed by immunoprecipitation and Western blotting. Migration capacity of primary CLL cells was analyzed by the Transwell migration assay. RESULTS: By analyzing the expression in 137 previously untreated CLL patients, we demonstrate that WNT5A and WNT5B genes show dramatically (five orders of magnitude) varying expression in CLL cells. High WNT5A and WNT5B expression strongly associates with unmutated IGHV and shortened time to first treatment. In addition, WNT5A levels associate, independent of IGHV status, with the clinically worst CLL subgroups characterized by dysfunctional p53 and mutated SF3B1. We provide functional evidence that WNT5A-positive primary CLL cells have increased motility and attenuated chemotaxis toward CXCL12 and CCL19 that can be overcome by inhibitors of Wnt/PCP signaling. CONCLUSIONS: These observations identify Wnt-5a as the crucial regulator of ROR1 activity in CLL and suggest that the autocrine Wnt-5a signaling pathway allows CLL cells to overcome natural microenvironmental regulation.

• MeSH
• autokrinní signalizace fyziologie   MeSH
• B-lymfocyty metabolismus   MeSH
• chemotaxe fyziologie   MeSH
• chronická lymfatická leukemie metabolismus   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• pohyb buněk fyziologie   MeSH
• proteiny Wnt metabolismus   MeSH
• protoonkogenní proteiny metabolismus   MeSH
• regulace genové exprese u nádorů fyziologie   MeSH
• signální dráha Wnt fyziologie   MeSH
• sirotčí receptory podobné receptoru tyrosinkinasy metabolismus   MeSH
• upregulace fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Intramural MeSH

Mammalian limb development is driven by the integrative input from several signaling pathways; a failure to receive or a misinterpretation of these signals results in skeletal defects. The brachydactylies, a group of overlapping inherited human hand malformation syndromes, are mainly caused by mutations in BMP signaling pathway components. Two closely related forms, Brachydactyly type B2 (BDB2) and BDB1 are caused by mutations in the BMP antagonist Noggin (NOG) and the atypical receptor tyrosine kinase ROR2 that acts as a receptor in the non-canonical Wnt pathway. Genetic analysis of Nog and Ror2 functional interaction via crossing Noggin and Ror2 mutant mice revealed a widening of skeletal elements in compound but not in any of the single mutants, thus indicating genetic interaction. Since ROR2 is a non-canonical Wnt co-receptor specific for Wnt-5a we speculated that this phenotype might be a result of deregulated Wnt-5a signaling activation, which is known to be essential for limb skeletal elements growth and patterning. We show that Noggin potentiates activation of the Wnt-5a-Ror2-Disheveled (Dvl) pathway in mouse embryonic fibroblast (MEF) cells in a Ror2-dependent fashion. Rat chondrosarcoma chondrocytes (RCS), however, are not able to respond to Noggin in this fashion unless growth arrest is induced by FGF2. In summary, our data demonstrate genetic interaction between Noggin and Ror2 and show that Noggin can sensitize cells to Wnt-5a/Ror2-mediated non-canonical Wnt signaling, a feature that in cartilage may depend on the presence of active FGF signaling. These findings indicate an unappreciated function of Noggin that will help to understand BMP and Wnt/PCP signaling pathway interactions.

• MeSH
• brachydaktylie genetika   MeSH
• genetické testování MeSH
• kostní morfogenetické proteiny antagonisté a inhibitory   MeSH
• lidé MeSH
• nervové dráhy MeSH
• protein Wnt 5a MeSH
• signální dráha Wnt MeSH
• signální transdukce * MeSH
• sirotčí receptory podobné receptoru tyrosinkinasy MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Nemalobuněčný karcinom plic (non-small cell lung cancer, NSCLC) tvoří 85 % všech karcinomů plic. Karcinom plic nadále zaujímá čelní příčky v příčinách úmrtí na nádorové onemocnění. V posledních dvou dekádách do terapie lokálně pokročilého nebo metastazujícího NSCLC vstoupila cílená léčba a imunoterapie, které změnily léčebné paradigma tohoto nádorového onemocnění. Obě léčebné modality vedly k významnému prodloužení celkového přežití pacientů s diagnózou pokročilého NSCLC.

Non-small cell lung carcinoma (NSCLC) accounts for 85% of all lung cancers. Lung cancer is one of the leading causes of cancer deaths. Due to targeted therapy and immunotherapy the treatment landscape for locally advanced or metastatic non-small-cell lung cancer has dramatically shifted over the past two decades. Both treatment modalities have significantly improved overall survival for patients with advanced NSCLC.

• MeSH
• anaplastická lymfomová kináza antagonisté a inhibitory   MeSH
• cílená molekulární terapie metody   MeSH
• geny erbB účinky léků   MeSH
• humanizované monoklonální protilátky terapeutické užití   MeSH
• imunoterapie * metody   MeSH
• klinické zkoušky, fáze III jako téma MeSH
• lidé MeSH
• nemalobuněčný karcinom plic * terapie   MeSH
• protinádorové látky imunologicky aktivní terapeutické užití   MeSH
• protoonkogeny účinky léků   MeSH
• sirotčí receptory podobné receptoru tyrosinkinasy terapeutické užití   MeSH
• Check Tag
• lidé MeSH

• MeSH
• anaplastická lymfomová kináza antagonisté a inhibitory   MeSH
• biologická terapie * metody   MeSH
• cílená molekulární terapie metody   MeSH
• genetické testování MeSH
• inhibitory angiogeneze terapeutické užití   MeSH
• inhibitory proteinkinas terapeutické užití   MeSH
• lidé MeSH
• monoklonální protilátky terapeutické užití   MeSH
• nádory plic * farmakoterapie   MeSH
• sirotčí receptory podobné receptoru tyrosinkinasy antagonisté a inhibitory   MeSH
• translokace genetická MeSH
• tyrosinkinasové receptory antagonisté a inhibitory   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

CAR T-lymfocyty dramaticky zasáhly do léčby CD19 pozitivních B-buněčných malignit. V tomto review jsme se zaměřili na použití CAR T-lymfocytů u hematologických malignit neexprimujících CD19, přičemž jsme zvláštní pozornost věnovali Hodgkinovu lymfomu a akutní myeloidní leukemii.

• MeSH
• akutní myeloidní leukemie imunologie   terapie   MeSH
• antigen CD33 antagonisté a inhibitory   MeSH
• antigen Ki-1 antagonisté a inhibitory   MeSH
• antigeny CD19 MeSH
• chimerické antigenní receptory * terapeutické užití   MeSH
• hematologické nádory imunologie   terapie   MeSH
• Hodgkinova nemoc imunologie   terapie   MeSH
• imunoterapie adoptivní * metody   MeSH
• lidé MeSH
• receptor interleukinu-3 - alfa-podjednotka antagonisté a inhibitory   MeSH
• receptory antigenů T-buněk antagonisté a inhibitory   MeSH
• sirotčí receptory podobné receptoru tyrosinkinasy antagonisté a inhibitory   MeSH
• Check Tag
• lidé MeSH

Mantle cell lymfom (mantle cell lymphoma, MCL) je vzácná B lymfoidní neoplazie, s mediánem věku v době iniciální diagnózy 68 let a s častějším výskytem u mužů. Základem současné léčby je chemoterapie v kombinaci s cílenou léčbou anti-CD20. Přestože se za posledních 30 let prognóza pacientů s MCL výrazně zlepšila s více než dvojnásobným přežitím, zůstává MCL pro většinu pacientů stále nevyléčitelné onemocnění. Průlom v léčbě relabovaných pacientů představují cílené inhibitory Brutonovy tyrosinkinázy. Současný vývoj a výzkum nabízí pacientům s MCL lepší perspektivu v podobě dalších nových léčiv, jako jsou bispecifické protilátky (CD20/CD3), anti-CD19 CAR-T buňky, nové protilátky (anti-CD19, anti-ROR1), imunokonjugáty (zilovertamab vedotin) a další léčiva.

Mantle cell lymphoma (MCL) is a rare B-lymphoid neoplasia, with a median 68 years at initial diagnosis and higher incidence among men. The recent therapy approach is based on chemotherapy combined with targeted anti-CD20. Despite a big improvement in the overall survival of patients with MCL, which is twice longer compared to that 30 years ago, MCL remains still incurable disease in majority of patients. In relapsing MCL patients, the inhibitors of Bruton's tyrosine kinases made a breakthrough. Recent development and research offer to MCL patients a better perspective in terms of new drugs: bispecific antibodies (CD20/CD3), anti-CD19 CAR-T, new antibodies or immunoconjugates (anti-CD19, anti-ROR1, zilovertamab vedotin) etc.

• MeSH
• antitumorózní látky terapeutické užití   MeSH
• chimerické antigenní receptory terapeutické užití   MeSH
• lidé MeSH
• lymfom z plášťových buněk * diagnóza   farmakoterapie   MeSH
• proteinkinasa BTK antagonisté a inhibitory   MeSH
• recidiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

In this review, we discuss the intricate roles of the Wnt signalling network in the development and progression of mature B-cell-derived haematological malignancies, with a focus on chronic lymphocytic leukaemia (CLL) and related B-cell lymphomas. We review the current literature and highlight the differences between the β-catenin-dependent and -independent branches of Wnt signalling. Special attention is paid to the role of the non-canonical Wnt/planar cell polarity (PCP) pathway, mediated by the Wnt-5-receptor tyrosine kinase-like orphan receptor (ROR1)-Dishevelled signalling axis in CLL. This is mainly because the Wnt/PCP co-receptor ROR1 was found to be overexpressed in CLL and the Wnt/PCP pathway contributes to numerous aspects of CLL pathogenesis. We also discuss the possibilities of therapeutically targeting the Wnt signalling pathways as an approach to disrupt the crucial interaction between malignant cells and their micro-environment. We also advocate the need for research in this direction for other lymphomas, namely, diffuse large B-cell lymphoma, Hodgkin lymphoma, mantle cell lymphoma, Burkitt lymphoma and follicular lymphoma where the Wnt signalling pathway probably plays a similar role. LINKED ARTICLES: This article is part of a themed section on WNT Signalling: Mechanisms and Therapeutic Opportunities. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.24/issuetoc.

• MeSH
• B-buněčný lymfom diagnóza   metabolismus   MeSH
• chronická lymfatická leukemie diagnóza   metabolismus   MeSH
• lidé MeSH
• signální dráha Wnt * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH