Genetics and prognostication in splenic marginal zone lymphoma: revelations from deep sequencing

PURPOSE: Mounting evidence supports the clinical significance of gene mutations and immunogenetic features in common mature B-cell malignancies.EXPERIMENTAL DESIGN: We undertook a detailed characterization of the genetic background of splenic marginal zone lymphoma (SMZL), using targeted re-sequencing and explored potential clinical implications in a multinational cohort of 175 SMZL patients.RESULTS: We identified recurrent mutations in TP53 (16%), KLF2 (12%), NOTCH2 (10%), TNFAIP3 (7%), MLL2 (11%), MYD88 (7%) and ARID1A (6%), all genes known to be targeted by somatic mutation in SMZL. KLF2 mutations were early, clonal events, enriched in patients with del(7q) and IGHV1-2*04 B-cell receptor immunoglobulins, and were associated with a short median time-to-first-treatment (0.12 vs. 1.11 yrs; P=0.01). In multivariate analysis mutations in NOTCH2 (HR 2.12, 95%CI 1.02-4.4, P=0.044) and 100% germline IGHV gene identity (HR 2.19, 95%CI 1.05-4.55, P=0.036) were independent markers of short time-to-first-treatment, while TP53 mutations were an independent marker of short overall survival (HR 2.36, 95% CI 1.08-5.2, P=0.03).CONCLUSIONS: We identify key associations between gene mutations and clinical outcome, demonstrating for the first time that NOTCH2 and TP53 gene mutations are independent markers of reduced treatment-free and overall survival, respectively.<br/

Cytogenetic complexity in chronic lymphocytic leukemia: definitions, associations and clinical impact

Recent evidence suggests that complex karyotype (CK) defined by the presence of 653 chromosomal aberrations (structural and/or numerical) identified by chromosome banding analysis (CBA) may be relevant for treatment decision-making in chronic lymphocytic leukemia (CLL). However, many challenges towards routine clinical application of CBA remain. In a retrospective study of 5290 patients with available CBA data, we explored both clinicobiological associations and the clinical impact of CK in CLL. We found that patients with 655 abnormalities, defined as high-CK, exhibit uniformly dismal clinical outcome, independently of clinical stage, TP53 aberrations (deletion of chromosome 17p and or TP53 mutations, TP53abs) and the expression of somatically hypermutated (M-CLL) or unmutated (U-CLL) immunoglobulin heavy variable genes (IGHV). Thus, they contrasted CK cases with 3 or 4 aberrations (low-CK and intermediate-CK, respectively) who followed aggressive disease courses only in the presence of TP53abs. At the other end of the spectrum, patients with CK and +12,+19 displayed an exceptionally indolent profile. Building upon CK, TP53abs and IGHV gene somatic hypermutation status, we propose a novel hierarchical model where patients with high-CK exhibit the worst prognosis, while M-CLL lacking CK or TP53abs as well as CK with +12,+19 show the longest overall survival. In conclusion, CK should not be axiomatically considered unfavorable in CLL, representing a heterogeneous group with variable clinical behavior. High-CK with 655 chromosomal aberrations emerges as prognostically adverse, independently of other biomarkers. Prospective clinical validation is warranted before finally incorporating high-CK in risk stratification of CLL

Disease-biased and shared characteristics of the immunoglobulin gene repertoires in marginal zone B cell lymphoproliferations.

The B cell receptor immunoglobulin (BcR IG) gene repertoires of marginal zone (MZ) lymphoproliferations were analyzed in order to obtain insight into their ontogenetic relationships. Our cohort included cases with MZ lymphomas (n=488) i.e. splenic (SMZL), nodal (NMZL) and extranodal (ENMZL) as well as provisional entities (n=76) according to the World Health Organization classification. The most striking IG gene repertoire skewing was observed in SMZL. However, restrictions were also identified in all other MZ lymphomas studied, particularly ENMZL, with significantly different IG gene distributions depending on the primary site of involvement. Cross-entity comparisons of the MZ IG sequence dataset with a large dataset of IG sequences (MZ-related or not; n=65,837) revealed four major clusters of cases sharing homologous ('public') heavy variable complementarity-determining region 3. These clusters included rearrangements from SMZL, ENMZL (gastric, salivary gland, ocular adnexa), chronic lymphocytic leukemia but also rheumatoid factors and non-malignant spleen MZ cells. In conclusion, different MZ lymphomas display biased immunogenetic signatures indicating distinct antigen exposure histories. The existence of rare public stereotypes raises the intriguing possibility that common, pathogen-triggered, immune-mediated mechanisms, may result in diverse B lymphoproliferations due to targeting versatile progenitor B cells and/or operating in particular microenvironments.This work was supported in part by H2020 “AEGLE, An analytics framework for integrated and personalized healthcare services in Europe”, by the European Union (EU); H2020 No. 692298 project “MEDGENET, Medical Genomics and Epigenomics Network” by the EU; grant AZV 15-30015A from the Ministry of Health of the Czech Republic, and the project CEITEC2020 LQ1601 from the Ministry of Education, Youth, and Sports of the Czech Republic; Bloodwise Research Grant (15019); the Swedish Cancer Society, the Swedish Research Council, the Knut and Alice Wallenberg Foundation, Karolinska Institutet, Stockholm, the Lion’s Cancer Research Foundation, Uppsala, the Marcus Borgström Foundation and Selander’s Foundation, Uppsala

The in vivo effect of glutamate ionotropic antagonists receptors in rat peripheral blood elements and bone marrow cells

Glutamate acid is one of the main endogenous amino acids in CNS. The purpose of this paper is to define the possible presense of ionotropic receptors in the hematopoietic system by administrating in vivo the ionotropic antagonists, MK-801 and NBQX, and glutamate agonists, MSG, in adult male rats. They were separated in three groups, one for peripheral blood studies and one for bone marrow studies and a control group. Studies were performed 12 and 24 hours after drug administration. Statistical analysis was done with Statistical Package SPSS 11.5 for windows. Based on the results, we came to the conclusion that glutamate ionotropic antagonists as well as glutamate itself, affect blood elements function and number, without any previous glutamate administration. Bone marrow function is also affected either as an adjustment to peripheral blood or as an effect of glutamate itself to bone marrow cells. More research is needed in the future in order to determine the exact mechanism of this affect and how we can use it in patient’s favour.Το γλουταμινικό οξύ είναι ένα από τα κύρια ενδογενή αμινοξέα του ΚΝΣ. Σκοπός της διατριβής αυτής είναι να ερευνήσει την παρουσία των NMDA υποδοχέων στο αιμοποιητικό σύστημα με την in vivo χορήγηση των ιονοτροπικών αναστολέων, ΜΚ801 και NBQX, και του άλατος του γλουταμινικού, MSG σε άρρενες επίμυες. Οι επίμυες χωρίστηκαν σε τρεις ομάδες, την ομάδα μελέτης του περιφερικού αίματος, την ομάδα μελέτης του μυελού των οστών και την ομάδα ελέγχου. Μετρήσεις γίνονταν 12 και 24 ώρες μετά τη χορήγηση των ουσιών. Η στατιστική ανάλυση των δεδομένων έγινε με τη βοήθεια του στατιστικού λογισμικού SPSS 11,5 for Windows. Με βάση τα αποτελέσματα της έρευνάς μας, διαπιστώνουμε ότι, τόσο οι ιονοτροπικοί ανταγωνιστές του γλουταμινικού οξέος, όσο και το ίδιο το άλας του γλουταμινικού οξέος, επηρεάζουν λειτουργικά και ποσοτικά τα έμμορφα στοιχεία του περιφερικού αίματος χωρίς να υπάρχει κάποια προηγούμενη διέγερση από το γλουταμινικό οξύ. Ως ένα σημείο, επηρεασμένη φαίνεται να είναι και η λειτουργία του μυελού των οστών. Περαιτέρω έρευνες θα χρειαστούν στο μέλλον ώστε να διαπιστωθεί ο ακριβής μηχανισμός της όποιας επίδρασης και το πώς η τελευταία μπορεί να χρησιμοποιηθεί προς όφελος του ασθενούς

How do Hematologists Communicate with Patients Suffering from Chronic Lymphocytic Leukemia? Insights from the ERIC Pilot Study in Greece

Chronic Lymphocytic Leukemia (CLL) is a chronic hematologic malignancy with great heterogeneity and unpredictable clinical course. The European Research Initiative on CLL (ERIC), in the context of its CLL Patient Empowerment Program, conducted a study exploring hematologists’ experience of communication. Thirty semi-structured interviews were conducted with hematologists caring for CLL patients in Greece. Inductive thematic qualitative analysis was employed revealing 3 major themes: (i) disclosure of information encompassing ‘negotiating the level of disclosure’, ‘the power of cultural perceptions’, ‘fear of being held culpable’, ‘fear of patients’ and own emotions’; (ii) medical-decision making which described ‘balancing autonomy and beneficence’, ‘considering patients’ preferences’ and ‘adhering to practice guidelines’; (iii) emotional support which included ‘assessment of emotional distress’, ‘identifying and regulating patients’ emotions’ and ‘maintaining a supportive relationship’. In conclusion, physicians are aware of the importance of communication and its potential impact on CLL patients. They use a wide range of communication strategies which serve the diversity of communication goals they must achieve. However, the majority have not received formal education on patient interactions. Present findings highlight the need for specific communication protocols, guidance and training that will empower physicians to overcome challenges, inherent to the nature of CLL

Integrating multiple immunogenetic data sources for feature extraction and mining somatic hypermutation patterns : the case of "towards analysis" in chronic lymphocytic leukaemia

Background: Somatic Hypermutation (SHM) refers to the introduction of mutations within rearranged V(D)J genes, a process that increases the diversity of Immunoglobulins (IGs). The analysis of SHM has offered critical insight into the physiology and pathology of B cells, leading to strong prognostication markers for clinical outcome in chronic lymphocytic leukaemia (CLL), the most frequent adult B-cell malignancy. In this paper we present a methodology for integrating multiple immunogenetic and clinocobiological data sources in order to extract features and create high quality datasets for SHM analysis in IG receptors of CLL patients. This dataset is used as the basis for a higher level integration procedure, inspired form social choice theory. This is applied in the Towards Analysis, our attempt to investigate the potential ontogenetic transformation of genes belonging to specific stereotyped CLL subsets towards other genes or gene families, through SHM. Results: The data integration process, followed by feature extraction, resulted in the generation of a dataset containing information about mutations occurring through SHM. The Towards analysis performed on the integrated dataset applying voting techniques, revealed the distinct behaviour of subset #201 compared to other subsets, as regards SHM related movements among gene clans, both in allele-conserved and non-conserved gene areas. With respect to movement between genes, a high percentage movement towards pseudo genes was found in all CLL subsets. Conclusions: This data integration and feature extraction process can set the basis for exploratory analysis or a fully automated computational data mining approach on many as yet unanswered, clinically relevant biological questions

Different Types of Chronic Inflammation Engender Distinctive Immunosenescent Profiles in Affected Patients

Immunosenescence encompasses a spectrum of lymphocyte phenotypic alterations. The aim of the study was to evaluate immunosenescent effect of two different forms of chronic inflammation, Systemic Lupus Erythematosous (SLE), a systemic autoimmune disease, and End-Stage Kidney Disease (ESKD), a chronic inflammatory disorder. Certain lymphocyte surface molecules, including CD31, CD45RA, CCR7, CD28, CD57, for T, and IgD, CD27 for B lymphocytes, were analyzed by flow cytometry in 30 SLE and 53 ESKD patients on hemodialysis (HD), and results were compared to 31 healthy controls (HC) of similar age, gender, and nationality. Significant Lymphopenia was evident in both SLE and ESKD-HD patients, compared to HC, affecting B cells 75.4 (14.4&ndash;520.8), 97 (32&ndash;341), and 214 (84&ndash;576) cells/&mu;L, respectively, p &lt; 0.0001, and CD4 cells 651.2 (71.1&ndash;1478.2), 713 (234&ndash;1509), and 986 (344&ndash;1591) cells/&mu;L, respectively, p &lt; 0.0001. The allocation of B cell subpopulations was remarkably different between SLE and ESKD-HD patients. SLE showed a clear shift to senescence (CD19IgD-CD27&minus;) cells, compared to ESKD-HD and HC, 11.75 (10)% vs. 8 (6) vs. 8.1 (10), respectively. Regarding T lymphocytes, Central Memory CD8 cells predominated in both SLE and ESKD-HD patients compared to HC, 53 (50)%, 52 (63), and 24 (64)%, respectively, while ESKD-HD but not SLE patients also had increased expression of CD4CD28&minus; and CD8CD28&minus; cells. In conclusion, both diseases are followed by significant lymphopenia; however, the senescent phenomenon affects the B lymphocyte compartment in SLE patients and T lymphocytes in ESKD-HD patients