Bernard-Soulier syndrome (BSS) is a rare inherited disorder characterized by unusually large platelets, low platelet count, and prolonged bleeding time. BSS is usually inherited in an autosomal recessive (AR) mode of inheritance due to a deficiency of the GPIb-IX-V complex also known as the von Willebrand factor (VWF) receptor. We investigated a family with macrothrombocytopenia, a mild bleeding tendency, slightly lowered platelet aggregation tests, and suspected autosomal dominant (AD) inheritance. We have detected a heterozygous GP1BA likely pathogenic variant, causing monoallelic BSS. A germline GP1BA gene variant (NM_000173:c.98G > A:p.C33Y), segregating with the macrothrombocytopenia, was detected by whole-exome sequencing. In silico analysis of the protein structure of the novel GPIbα variant revealed a potential structural defect, which could impact proper protein folding and subsequent binding to VWF. Flow cytometry, immunoblot, and electron microscopy demonstrated further differences between p.C33Y GP1BA carriers and healthy controls. Here, we provide a detailed insight into its clinical presentation and phenotype. Moreover, the here described case first presents an mBSS patient with two previous ischemic strokes.

• Klíčová slova
• Bernard-Soulier syndrome, GP1BA, autosomal dominant, macrothrombocytopenia, monoallelic,
• MeSH
• alely * MeSH
• Bernardův-Soulierův syndrom krev   diagnóza   genetika   MeSH
• fenotyp * MeSH
• genetická predispozice k nemoci * MeSH
• genetická variace * MeSH
• genetické asociační studie MeSH
• imunofenotypizace MeSH
• lidé MeSH
• mutační analýza DNA MeSH
• počet trombocytů MeSH
• rodokmen MeSH
• trombocytopenie krev   diagnóza   MeSH
• trombocytový glykoproteinový komplex Ib-IX genetika   metabolismus   MeSH
• trombocyty metabolismus   ultrastruktura   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• adhesion receptor MeSH Prohlížeč
• trombocytový glykoproteinový komplex Ib-IX MeSH

No data are available regarding obesity and outcome in Chronic Lymphocytic Leukemia (CLL). We analyzed 263 patients from the AGMT CLL-8a Mabtenance trial for the impact of obesity. The trial included patients after rituximab-containing induction treatment in first or second line that had achieved at least a PR. A randomization to rituximab maintenance treatment (375 mg/m2 q3 months for 2 years) vs observation was performed. In this cohort 22% of the patients (58/263) were classified as obese. The baseline response to induction treatment was inferior in obese patients with a lower CR rate (43.1% vs 60.5% in obese vs non-obese, P = 0.018) and with a lower rate of patients achieving MRD negativity after chemoimmunotherapy induction treatment (19.6% vs 35.8%, P = 0.02). The PFS outcome of obese patients was significantly worse in the observation group of the trial (24 vs 39 months median PFS, P = 0.03). However, in the rituximab maintenance group the outcome for obese vs non-obese was not different (P = 0.4). In summary, obesity was overall associated with a worse outcome of chemoimmunotherapy induction. However, rituximab maintenance treatment seems to be able to overcome this negative effect.

• Klíčová slova
• BMI, CLL, maintenance, obesity, rituximab,
• MeSH
• analýza přežití MeSH
• chronická lymfatická leukemie farmakoterapie   mortalita   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• obezita komplikace   mortalita   MeSH
• prognóza MeSH
• rituximab aplikace a dávkování   terapeutické užití   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• udržovací chemoterapie MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• randomizované kontrolované studie MeSH
• Názvy látek
• rituximab MeSH

Mutations in the GP1BA gene have been associated with platelet-type von Willebrand disease and Bernard-Soulier syndrome. Here, we report a novel GP1BA mutation in a family with autosomal dominant macrothrombocytopenia and mild bleeding. We performed analyses of seven family members. Using whole-exome sequencing of germline DNA samples, we identified a heterozygous single-nucleotide change in GP1BA (exone2:c.176T>G), encoding a p.Leu59Arg substitution in the N-terminal domain, segregating with macrothrombocytopenia. This variant has not been previously reported. We also analysed the structure of the detected sequence variant in silico. In particular, we used the crystal structure of the human platelet receptor GP Ibα N-terminal domain. Replacement of aliphatic amino-acid Leu 59 with charged, polar and larger arginine probably disrupts the protein structure. An autosomal dominant mode of inheritance, a family history of mild bleeding episodes, aggregation pattern in affected individuals together with evidence of mutation occurring in part of the GP1BA gene encoding the leucine-rich repeat region suggest a novel variant causing monoallelic Bernard-Soulier syndrome.

• Klíčová slova
• Autosomal dominant variant, GP1BA, Inherited thrombocytopenia, monoallelic Bernard-Soulier syndrome,
• MeSH
• Bernardův-Soulierův syndrom genetika   metabolismus   MeSH
• bodová mutace * MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• proteinové domény MeSH
• trombocytový glykoproteinový komplex Ib-IX chemie   genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Názvy látek
• adhesion receptor MeSH Prohlížeč
• trombocytový glykoproteinový komplex Ib-IX MeSH

BACKGROUND: Minimal residual disease (MRD) is an important prognostic maker in acute lymphoblastic leukemia (ALL). However, few data comparing the measurement of adult ALL MRD using different methods in daily practice are available. We conducted an analysis comparing the importance of flow cytometry (FCM) and real-time quantitative polymerase chain reaction (PCR) in the assessment of MRD in adult ALL. PATIENTS AND METHODS: Fifty-six consecutive adult patients with both Philadelphia-negative and -positive ALL treated according to an intensive protocol were enrolled in the study. Bone marrow samples were acquired on day 26 and during week 11 of treatment. MRD evaluation was performed using 8-color FCM and PCR of immunoglobulin or T-cell receptor gene clonal rearrangements and BCR-ABL1, KMT2A-AF4 and E2A-PBX1 fusion genes. RESULTS: On day 26, both FCM and PCR seemed to have good discrimination sensitivity for overall survival (P = .001 to .008) and progression-free survival (P = .03 to .04) prediction for both Philadelphia-positive and -negative cases. The most sensitive method in week 11 was PCR including all results > 0 considered to indicate MRD positivity (P = .002 for overall survival and P = .02 for progression-free survival). PCR with other cutoffs was not sufficiently sensitive in week 11. Moreover, no FCM+ samples were found in week 11. The subanalysis of the Philadelphia-negative patients showed similar results. CONCLUSION: Our analysis showed that both FCM and PCR MRD assessment methods are sensitive for survival prediction during induction. However, we believe FCM could not be sufficiently sensitive in later phases of treatment.

• Klíčová slova
• ALL, FCM, MRD, Overall survival, Sensitivity,
• MeSH
• akutní lymfatická leukemie terapie   MeSH
• dospělí MeSH
• kostní dřeň MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• míra přežití MeSH
• mladiství MeSH
• mladý dospělý MeSH
• následné studie MeSH
• prognóza MeSH
• průtoková cytometrie metody   MeSH
• retrospektivní studie MeSH
• reziduální nádor diagnóza   etiologie   metabolismus   MeSH
• senioři MeSH
• transplantace kmenových buněk škodlivé účinky   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

The diagnostic criteria for CLL rely on morphology and immunophenotype. Current approaches have limitations affecting reproducibility and there is no consensus on the role of new markers. The aim of this project was to identify reproducible criteria and consensus on markers recommended for the diagnosis of CLL. ERIC/ESCCA members classified 14 of 35 potential markers as "required" or "recommended" for CLL diagnosis, consensus being defined as >75% and >50% agreement, respectively. An approach to validate "required" markers using normal peripheral blood was developed. Responses were received from 150 participants with a diagnostic workload >20 CLL cases per week in 23/150 (15%), 5-20 in 82/150 (55%), and <5 cases per week in 45/150 (30%). The consensus for "required" diagnostic markers included: CD19, CD5, CD20, CD23, Kappa, and Lambda. "Recommended" markers potentially useful for differential diagnosis were: CD43, CD79b, CD81, CD200, CD10, and ROR1. Reproducible criteria for component reagents were assessed retrospectively in 14,643 cases from 13 different centers and showed >97% concordance with current approaches. A pilot study to validate staining quality was completed in 11 centers. Markers considered as "required" for the diagnosis of CLL by the participants in this study (CD19, CD5, CD20, CD23, Kappa, and Lambda) are consistent with current diagnostic criteria and practice. Importantly, a reproducible approach to validate and apply these markers in individual laboratories has been identified. Finally, a consensus "recommended" panel of markers to refine diagnosis in borderline cases (CD43, CD79b, CD81, CD200, CD10, and ROR1) has been defined and will be prospectively evaluated. © 2017 International Clinical Cytometry Society.

• Klíčová slova
• chronic lymphocytic leukemia, diagnosis, flow cytometry,
• MeSH
• chronická lymfatická leukemie diagnóza   metabolismus   MeSH
• imunofenotypizace metody   MeSH
• lidé MeSH
• nádorové biomarkery metabolismus   MeSH
• pilotní projekty MeSH
• průtoková cytometrie metody   MeSH
• reprodukovatelnost výsledků MeSH
• retrospektivní studie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• nádorové biomarkery MeSH

• Klíčová slova
• ROR1, cell isolation, chronic lymphocytic leukaemia, complete remission,
• 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
• Názvy látek
• ROR1 protein, human MeSH Prohlížeč
• sirotčí receptory podobné receptoru tyrosinkinasy MeSH

BACKGROUND: In many patients with chronic lymphocytic leukaemia requiring treatment, induction therapy with rituximab plus chemotherapy improves outcomes compared with chemotherapy alone. In this study we aimed to investigate the potential of rituximab maintenance therapy to prolong disease control in patients who respond to rituximab-containing induction regimens. METHODS: In this randomised, international, multicentre, open-label, phase 3 clinical trial, we enrolled patients who had achieved a complete response (CR), CR with incomplete bone marrow recovery (CRi), or partial response (PR) to first-line or second-line rituximab-containing chemoimmunotherapy and randomly assigned them in a 1:1 ratio (central block randomisation in the electronic case report form system) to either intravenous rituximab 375 mg/m(2) every 3 months, or observation alone, for 2 years. Stratification was by country, line of treatment, type of chemotherapy added to the rituximab backbone, and degree of remission following induction. The primary endpoint was progression-free survival. Efficacy analysis was done in the intention-to-treat population. This is the final, event-triggered analysis. Final analysis was triggered by the occurrence of 92 events. This trial is registered with ClinicalTrials.gov, number NCT01118234. FINDINGS: Between April 1, 2010, and Dec 23, 2013, 134 patients were randomised to rituximab and 129 to observation alone. Median observation times were 33·4 months (IQR 25·7-42·8) for the rituximab group and 34·0 months (25·4-41·9) for the observation group. Progression-free survival was significantly longer in the rituximab maintenance group (47·0 months, IQR 28·5-incalculable) than with observation alone (35·5 months, 95% CI 25·7-46·3; hazard ratio [HR] 0·50, 95% CI 0·33-0·75, p=0·00077). The incidence of grade 3-4 haematological toxicities other than neutropenia was similar in the two treatment groups. Grade 3-4 neutropenia occurred in 28 (21%) patients in the rituximab group and 14 (11%) patients in the observation group. Apart from neutropenia, the most common grade 3-4 adverse events were upper (five vs one [1%] patient in the observation group) and lower (three [2%] vs one [1%]) respiratory tract infection, pneumonia (nine [7%] vs two [2%]), thrombopenia (four [3%] vs four [3%]), neoplasms (five [4%] vs four [3%]), and eye disorders (four [3%] vs two [2%]). The overall incidence of infections of all grades was higher among rituximab recipients (88 [66%] vs 65 [50%]). INTERPRETATION: Rituximab maintenance therapy prolongs progression-free survival in patients achieving at least a PR to induction with rituximab plus chemotherapy, and the treatment is well tolerated overall. Although it is associated with an increase in infections, there is no excess in infection mortality, suggesting that remission maintenance with rituximab is an effective and safe option in the management of chronic lymphocytic leukaemia in early treatment phases. FUNDING: Arbeitsgemeinschaft Medikamentöse Tumortherapie gemeinnützige GmbH (AGMT), Roche.

• MeSH
• bendamustin hydrochlorid aplikace a dávkování   MeSH
• chronická lymfatická leukemie farmakoterapie   patologie   patofyziologie   MeSH
• cyklofosfamid aplikace a dávkování   MeSH
• dospělí MeSH
• imunoterapie * MeSH
• indukce remise MeSH
• lidé středního věku MeSH
• lidé MeSH
• míra přežití MeSH
• protokoly protinádorové kombinované chemoterapie aplikace a dávkování   terapeutické užití   MeSH
• rituximab aplikace a dávkování   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• staging nádorů MeSH
• vidarabin aplikace a dávkování   analogy a deriváty   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky, fáze III MeSH
• multicentrická studie MeSH
• randomizované kontrolované studie MeSH
• srovnávací studie MeSH
• Názvy látek
• bendamustin hydrochlorid MeSH
• cyklofosfamid MeSH
• fludarabine MeSH Prohlížeč
• rituximab MeSH
• vidarabin 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
• protein Wnt 5a 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
• Názvy látek
• protein Wnt 5a MeSH
• proteiny Wnt MeSH
• protoonkogenní proteiny MeSH
• ROR1 protein, human MeSH Prohlížeč
• sirotčí receptory podobné receptoru tyrosinkinasy MeSH
• WNT5A protein, human MeSH Prohlížeč
• WNT5B protein, human MeSH Prohlížeč

The treatment of relapsed/refractory chronic lymphocytic leukemia (CLL) remains a challenging clinical issue. An important treatment option is the use of high-dose corticosteroids. The purpose of this clinical trial was to determine the efficacy and toxicity of an ofatumumab-dexamethasone (O-Dex) combination in relapsed or refractory CLL. The trial was an open-label, multicenter, nonrandomized, Phase II study. The O-Dex regimen consisted of intravenous ofatumumab (Cycle 1: 300 mg on day 1, 2,000 mg on days 8, 15, and 22; Cycles 2-6: 1,000 mg on days 1, 8, 15, and 22) and oral dexamethasone (40 mg on days 1-4 and 15-18; Cycles 1-6). The O-Dex regimen was given until best response, or a maximum of six cycles. Thirty-three patients (pts) were recruited. Twenty-four (73%) pts completed at least three cycles of therapy. The remaining nine pts were prematurely discontinued owing to Grade 3/4 infections (seven pts), disease progression (one pt), or uncontrollable diabetes mellitus (one pt). Overall response rates/complete remissions (ORR/CR) were achieved in 22/5 pts (67/15%). The median progression-free survival (PFS) was 10 months. In pts with p53 defects (n = 8), ORR/CR were achieved in 5/2 pts (63/25%) with a median PFS of 10.5 months. The median overall survival (OS) was 34 months. The Grades 3-5 infectious toxicity in 33% of pts represented the most frequent side effect during the treatment period. In conclusion, the O-Dex regimen shows a relatively high ORR and CR with promising findings for PFS and OS. The study was registered at www.clinicaltrials.gov (NCT01310101).

• MeSH
• analýza přežití MeSH
• chronická lymfatická leukemie farmakoterapie   genetika   mortalita   patologie   MeSH
• dexamethason terapeutické užití   MeSH
• humanizované monoklonální protilátky MeSH
• kombinovaná farmakoterapie metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• monoklonální protilátky terapeutické užití   MeSH
• mutace MeSH
• nádorový supresorový protein p53 genetika   MeSH
• protinádorové látky terapeutické užití   MeSH
• recidiva MeSH
• rozvrh dávkování léků MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky, fáze II MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dexamethason MeSH
• humanizované monoklonální protilátky MeSH
• monoklonální protilátky MeSH
• nádorový supresorový protein p53 MeSH
• ofatumumab MeSH Prohlížeč
• protinádorové látky MeSH

BACKGROUND: Minimal residual disease (MRD) detection is an essential tool for therapy response assessment in a considerable number of hematooncologic disorders, including mantle cell lymphoma (MCL). Flow cytometry (FCM) ranks among the most effective approaches, which allows rapid sample processing and compete successfully with highly sensitive molecular methods like polymerase chain reaction. Because FCM is ordinarily applied to detect MRD in B-lineage diseases like chronic lymphocytic leukemia, a similar method could be used in MCL. We decided to test our novel eight-color FCM approach in MCL MRD detection. METHODS: Using an eight-color FCM protocol designed by us, the expression of 24 selected surface antigens in a cohort of 30 patients with newly diagnosed leukemic MCL and 20 normal controls were compared to establish markers that can reliably distinguish normal B-lymphocytes from the MCL population. The sensitivity of the designed protocol was tested using serial dilution studies. RESULTS: Although MCL presents high immunophenotypic variability, the combination of CD20/23/5/19/200/62L/3/45 seems to be very favorable in flow cytometric MRD measurement. CONCLUSIONS: Our eight-color FCM protocol could easily detect MRD in MCL patients, and reaches a sensitivity of up to 2 × 10(-4) .

• Klíčová slova
• flow cytometry, mantle cell lymphoma, minimal residual disease,
• MeSH
• dospělí MeSH
• kohortové studie MeSH
• lidé středního věku MeSH
• lidé MeSH
• lymfom z plášťových buněk krev   diagnóza   MeSH
• průtoková cytometrie metody   normy   MeSH
• reziduální nádor krev   diagnóza   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH