Cannabinoid receptors expression in bone marrow trephine biopsy of chronic lymphocytic leukaemia patients treated with purine analogues

Background: Cannabinoid receptors CB1 and CB2 are part the endocannabinoid system that plays an important role in the process of proliferation and apoptosis of different neoplastic cells. B-cell chronic lymphocytic leukaemia is one of the diseases in which these processes are altered. Aim: The aim of our study was the assessment of cannabinoid receptor expression on the B-lymphocytes in bone marrow trephine biopsy from leukaemic patients at diagnosis and after purine analogue treatment. Methods: The biopsy was taken routinely and standard immunohistochemical staining procedure for paraffin embedded sections was applied. The cannabinoid receptors were detected using specific primary polyclonal antibody anti-CB1 and anti-CB2. Additionally, an existence of cannabinoid receptors was confirmed by flow cytometry. Results: The results showed that the expression of CB1 receptor on the surface of neoplastic cells was lower than that of CB2 (17.0 ± 3.1% and 92.1 ± 1.7% respectively, p < 0.001). Nine of the patients responded to applied treatment with a reduction in leukaemic infiltration (77.2 ± 6.9% to 30.2 ± 6.5%, p = 0.007) and CB1 receptor expression (24.4 ± 4.8% to 8.6 ± 2.9%, p = 0.01), but there was no change in CB2 expression (91.7 ± 2.7% vs 90.9 ± 2.8%, p = 0.69). Four patients without remission expressed even greater number of the receptors. In all of the cases both cannabinoid receptor types antibodies gave positive reaction. Furthermore, the existence of cannabinoid receptors on neoplastic lymphocytes was confirmed by flow cytometry. Conclusion: The study provides original evidence for the existence of cannabinoid receptors on B-lymphocytes in chronic lymphocytic leukaemia patients. The receptors are thought to be a new structure that can modify the course of the disease and may be considered as a new target in leukaemia treatment.Обоснование: рецепторы каннабиноидов CB1 и CB2 яв ляются частью системы эндоканнабиноидов, которая играет важную роль в процессах пролиферации и апоптоза различных неопластических клеток. Одним из заболеваний, при которых происходит нарушение этих процессов, является В-клеточный хронический лимфлейкоз. Цель: оценка экспрессии рецепторов каннабиноидов на В-лимфоцитах в трепанобиоптатах у больных лейкозом до и после проведения лечения с использованием пуриновых аналогов. Методы: биоптаты получили рутинными методами; иммуногистохимическое исследование депарафинизированных срезов проводили по стандартной процедуре. Рецепторы каннабиноидов определяли с использованием специфических моноклональных антител анти-CB1 и анти-CB2. Кроме того, наличие рецепторов подтверждено при по- мощи проточной цитофлуориметрии. Результаты: показано, что экспрессия рецептора CB1 на поверхности опухолевых клеток ниже, чем экспрессия CB2 (17,0  ± 3,1% и 92,1 1,7% твенно, p < 0,001). После проведенного лечения у 9 пациентов отмечалось уменьшение лейкозного инфильтрата (77,2 6,9% до 30,2 6,5%, p = 0,007) и снижение экспрессии рецептора CB1 (24,4 4,8% до 8,6 2,9%, p = 0,01), однако различий в экспрессии CB2 не отмечали (91,7 2,7% против 90,9 2,8%, p = 0,69). У 4 пациентов, у которых не удалось достичь ремиссии, определяли даже повышение экспрессии рецепторов. Во всех случаях маркировки антителами к обоим типам рецепторов каннабиноидов отмечали поло- жительную реакцию. Более того, присутствие рецепторов каннабиноидов на злокачественных клетках подтверждали при помощи проточной цитометрии. Выводы: в ходе исследования показано изначальное наличие рецепторов каннабиноидов на В-лимфоцитах у больных хроническим лимфолейкозом. Указанные рецепторы могут быть новой структурой, которая может быть модифицирована в течение болезни, и могут считаться новой мишенью при лечении больных лейкозом

Syntheses, characterization, density functional theory calculations, and activity of tridentate SNS zinc pincer complexes

A series of tridentate SNS ligand precursors were metallated with ZnCl2 to give new tridentate SNS pincer zinc complexes. The zinc complexes serve as models for the zinc active site in liver alcohol dehydrogenase (LADH) and were characterized with single crystal X-ray diffraction, 1H, 13C, and HSQC NMR spectroscopies and electrospray mass spectrometry. The bond lengths and bond angles of the zinc complexes correlate well to those in horse LADH. The zinc complexes feature SNS donor atoms and pseudotetrahedral geometry about the zinc center, as is seen for liver alcohol dehydrogenase. The SNS ligand precursors were characterized with 1H, 13C, and HSQC NMR spectroscopies and cyclic voltammetry, and were found to be redox active. Gaussian calculations were performed and agree quite well with the experimentally observed oxidation potential for the pincer ligand. The zinc complexes were screened for the reduction of electron poor aldehydes in the presence of a hydrogen donor, 1-benzyl-1,4-dihydronicotinamide (BNAH). The zinc complexes enhance the reduction of electron poor aldehydes. Density functional theory calculations were performed to better understand why the geometry about the zinc center is pseudo-tetrahedral rather than pseudo-square planar, which is seen for most pincer complexes. For the SNS tridentate pincer complexes, the data indicate that the pseudo-tetrahedral geometry was 43.8 kcal/mol more stable than the pseudo-square planar geometry. Density functional theory calculations were also performed on zinc complexes with monodentate ligands and the data indicate that the pseudo-tetrahedral geometry was 30.6 kcal/mol more stable than pseudo-square planar geometry. Overall, the relative stabilities of the pseudo-tetrahedral and pseudo-square planar systems are the same for this coordination environment whether the ligand set is a single tridentate SNS system or is broken into three separate units. The preference of a d10 Zn center to attain a tetrahedral local environment trumps any stabilization gained by removal of constraints within the ligand set

Syntheses, Characterization, Density Functional Theory Calculations, and Activity of Tridentate SNS Zinc Pincer Complexes Based on Bis-Imidazole or Bis-Triazole Precursors

A series of tridentate pincer ligands, each possessing two sulfur- and one nitrogen-donor functionalities (SNS), based on bis-imidazole or bis-triazole salts were metallated with ZnCl2 to give new tridentate SNS pincer zinc(II) complexes [(SNS)ZnCl]+. The zinc complexes serve as models for the zinc active site in liver alcohol dehydrogenase (LADH) and were characterized with single crystal X-ray diffraction, 1H, 13C, and HSQC NMR spectroscopies, electrospray mass spectrometry, and elemental analysis. The zinc complexes feature SNS donor atoms and pseudotetrahedral geometry about the zinc center, as is seen for liver alcohol dehydrogenase. The bond lengths and bond angles of the zinc complexes correlate well to those in horse LADH. The SNS ligand precursors were characterized with 1H, 13C, and HSQC NMR spectroscopies, elemental analysis, and cyclic voltammetry, and were found to be redox active. Gaussian calculations were performed and agree with the experimentally observed oxidation potentials for the pincer ligand precursors. The zinc complexes were screened for the reduction of electron-poor aldehydes in the presence of a hydrogen donor, 1-benzyl-1,4-dihydronicotinamide (BNAH), and it was determined that they enhance the reduction of electron-poor aldehydes. The SNS zinc pincer complexes with bis-triazole ligand precursors exhibit higher activity for the reduction of 4-nitrobenzaldehyde than do SNS zinc pincer complexes with bis-imidazole ligand precursors. Quantitative stoichiometric conversion was seen for the reduction of pyridine-2-carboxaldehyde via SNS zinc pincer complexes with either bis-imidazole or bis-triazole ligand precursors

Gene Targeting to the Uteroplacental Circulation of Pregnant Guinea Pigs

Our study aimed to target adenoviral gene therapy to the uteroplacental circulation of pregnant guinea pigs in order to develop a novel therapy for fetal growth restriction. Four methods of delivery of an adenovirus encoding β-galactosidase (Ad.LacZ) were evaluated: intravascular injection using phosphate-buffered saline (PBS) into (1) uterine artery (UtA) or (2) internal iliac artery or external administration in (3) PBS or (4) pluronic F-127 gel (Sigma Aldrich). Postmortem examination was performed 4 to 7 days after gene transfer. Tissue transduction was assessed by X-gal histochemistry and enzyme-linked immunosorbent assay. External vascular application of the adenovirus vector in combination with pluronic gel had 91.7% success rate in terms of administration (85% maternal survival) and gave the best results for maternal/fetal survival and local transduction efficiency without any spread to maternal or fetal tissues. This study suggests an optimal method of gene delivery to the UtAs of a small rodent for preclinical studies

Multiobjective Design Optimization using Nash Games

International audienceIn the area of pure numerical simulation of multidisciplinary coupled systems, the computational cost to evaluate a configuration may be very high. A fortiori, in multi- disciplinary optimization, one is led to evaluate a number of different configurations to iterate on the design parameters. This observation motivates the search for the most in- novative and computationally efficient approaches in all the sectors of the computational chain : at the level of the solvers (using a hierarchy of physical models), the meshes and geometrical parameterizations for shape, or shape deformation, the implementation (on a sequential or parallel architecture; grid computing), and the optimizers (deterministic or semi-stochastic, or hybrid; synchronous, or asynchronous). In the present approach, we concentrate on situations typically involving a small number of disciplines assumed to be strongly antagonistic, and a relatively moderate number of related objective functions. However, our objective functions are functionals, that is, PDE-constrained, and thus costly to evaluate. The aerodynamic and structural optimization of an aircraft configuration is a prototype of such a context, when these disciplines have been reduced to a few major objectives. This is the case when, implicitly, many subsystems are taken into account by local optimizations. Our developments are focused on the question of approximating the Pareto set in cases of strongly-conflicting disciplines. For this purpose, a general computational technique is proposed, guided by a form of sensitivity analysis, with the additional objective to be more economical than standard evolutionary approaches

Zanubrutinib for the treatment of MYD88 wild-type Waldenström macroglobulinemia: a substudy of the phase 3 ASPEN trial

Abstract Patients with Waldenström macroglobulinemia (WM) lacking activating mutations in the MYD88 gene (MYD88WT) have demonstrated relatively poor outcomes to ibrutinib monotherapy, with no major responses reported in a phase 2 pivotal study. Zanubrutinib is a novel, selective Bruton tyrosine kinase (BTK) inhibitor designed to maximize BTK occupancy and minimize off-target activity. The ASPEN study consisted of a randomized comparison of zanubrutinib and ibrutinib efficacy and safety in patients with WM who have the MYD88 mutation, as well as a separate cohort of patients without MYD88 mutation (MYD88WT) or with unknown mutational status who received zanubrutinib. Results from the latter single-arm cohort are reported herein. Efficacy endpoints included overall, major and complete (CR) or very good partial response (VGPR) rates, progression-free survival (PFS), duration of response (DOR), and overall survival (OS). Twenty-eight patients (23 relapsed/refractory; 5 treatment-naïve) were enrolled, including 26 with centrally confirmed MYD88WT disease and 2 with unknown MYD88 mutational status. At a median follow-up of 17.9 months, 7 of 26 MYD88WT patients (27%) had achieved a VGPR and 50% a major response (partial response or better); there were no CRs. At 18 months, the estimated PFS and OS rates were 68% and 88%, respectively, while the median DOR had not been reached. Two patients discontinued zanubrutinib due to adverse events. Treatment-emergent hypertension, atrial fibrillation, and major hemorrhages were reported in 3, 1 and 2 patients (including 1 concurrent with enoxaparin therapy), respectively. Results of this substudy demonstrate that zanubrutinib monotherapy can induce high quality responses in patients with MYD88WT WM. This trial is registered on www.clinicaltrials.gov as NCT #03053440.</jats:p