miR-217 is an oncogene that enhances the germinal center reaction.

microRNAs are a class of regulators of gene expression that have been shown critical for a great number of biological processes; however, little is known of their role in germinal center (GC) B cells. Although the GC reaction is crucial to ensure a competent immune response, GC B cells are also the origin of most human lymphomas, presumably due to bystander effects of the immunoglobulin gene remodeling that takes place at these sites. Here we report that miR-217 is specifically upregulated in GC B cells. Gain- and loss-of-function mouse models reveal that miR-217 is a positive modulator of the GC response that increases the generation of class-switched antibodies and the frequency of somatic hypermutation. We find that miR-217 down-regulates the expression of a DNA damage response and repair gene network and in turn stabilizes Bcl-6 expression in GC B cells. Importantly, miR-217 overexpression also promotes mature B-cell lymphomagenesis; this is physiologically relevant as we find that miR-217 is overexpressed in aggressive human B-cell lymphomas. Therefore, miR-217 provides a novel molecular link between the normal GC response and B-cell transformation.

PLCG1 mutations in cutaneous T-cell lymphomas.

Cutaneous T-cell lymphoma (CTCL) is a heterogeneous group of primary cutaneous T-cell lymphoproliferative processes, mainly composed of mycosis fungoides and Sézary syndrome, the aggressive forms of which lack an effective treatment. The molecular pathogenesis of CTCL is largely unknown, although neoplastic cells show increased signaling from T-cell receptors (TCRs). DNAs from 11 patients with CTCL, both normal and tumoral, were target-enriched and sequenced by massive parallel sequencing for a selection of 524 TCR-signaling-related genes. Identified variants were validated by capillary sequencing. Multiple mutations were found that affected several signaling pathways, such as TCRs, nuclear factor κB, or Janus kinase/signal transducer and activator of transcription, but PLCG1 was found to be mutated in 3 samples, 2 of which featured a redundant mutation (c.1034T>C, S345F) in exon 11 that affects the PLCx protein catalytic domain. This mutation was further analyzed by quantitative polymerase chain reaction genotyping in a new cohort of 42 patients with CTCL, where it was found in 19% of samples. Immunohistochemical analysis for nuclear factor of activated T cells (NFAT) showed that PLCG1-mutated cases exhibited strong NFAT nuclear immunostaining. Functional studies demonstrated that PLCG1 mutants elicited increased downstream signaling toward NFAT activation, and inhibition of this pathway resulted in reduced CTCL cell proliferation and cell viability. Thus, increased proliferative and survival mechanisms in CTCL may partially depend on the acquisition of somatic mutations in PLCG1 and other genes that are essential for normal T-cell differentiation.

NF-κB directly mediates epigenetic deregulation of common microRNAs in Epstein-Barr virus-mediated transformation of B-cells and in lymphomas.

MicroRNAs (miRNAs) have negative effects on gene expression and are major players in cell function in normal and pathological conditions. Epstein-Barr virus (EBV) infection of resting B lymphocytes results in their growth transformation and associates with different B cell lymphomas. EBV-mediated B cell transformation involves large changes in gene expression, including cellular miRNAs. We performed miRNA expression analysis in growth transformation of EBV-infected B cells. We observed predominant downregulation of miRNAs and upregulation of a few miRNAs. We observed similar profiles of miRNA expression in B cells stimulated with CD40L/IL-4, and those infected with EBNA-2- and LMP-1-deficient EBV particles, suggesting the implication of the NF-kB pathway, common to all four situations. In fact, the NF-kB subunit p65 associates with the transcription start site (TSS) of both upregulated and downregulated miRNAs following EBV infection This occurs together with changes at histone H3K27me3 and histone H3K4me3. Inhibition of the NF-kB pathway impairs changes in miRNA expression, NF-kB binding and changes at the above histone modifications near the TSS of these miRNA genes. Changes in expression of these miRNAs also occurred in diffuse large B cell lymphomas (DLBCL), which are strongly NF-kB dependent. Our results highlight the relevance of the NF-kB pathway in epigenetically mediated miRNA control in B cell transformation and DLBCL.

The role of miRNAs in the pathogenesis and diagnosis of B-cell lymphomas.

There is a demand to understand B-cell lymphoma pathogenesis better, to identify new markers, and to define multiple lymphoproliferative disorders more accurately. MicroRNAs (miRNAs) are regulators of protein translation, comprising a group of more than 1500 short noncoding single-strand RNA molecules of approximately 22 nucleotides in length. They are easily detectable in fresh or paraffin-embedded diagnostic tissue and serum. Expression of individual miRNAs and miRNA signatures allows specific cell-differentiation stages to be identified, and is a powerful diagnostic and prognostic method. Here we review what is known about the pathogenic relevance of miRNAs, and use of miRNAs for the diagnosis and prognosis of B-cell lymphomas. Most of the published data concern chronic lymphocytic lymphoma and diffuse large B-cell lymphoma, and implicate miRNAs in the pathogenesis of these diseases. They identify miRNAs that could be used for diagnosis, prognosis, or prediction of response to specific therapies.

Nodal marginal zone lymphoma: gene expression and miRNA profiling identify diagnostic markers and potential therapeutic targets.

Nodal marginal zone lymphoma (NMZL) is a small B-cell neoplasm whose molecular pathogenesis is still essentially unknown and whose differentiation from other small B-cell lymphomas is hampered by the lack of specific markers. We have analyzed gene expression, miRNA profile, and copy number data from 15 NMZL cases. For comparison, 16 follicular lymphomas (FLs), 9 extranodal marginal zone lymphomas, and 8 reactive lymph nodes and B-cell subtypes were included. The results were validated by quantitative RT-PCR in an independent series, including 61 paraffin-embedded NMZLs. NMZL signature showed an enriched expression of gene sets identifying interleukins, integrins, CD40, PI3K, NF-κB, and TGF-ß, and included genes expressed by normal marginal zone cells and memory B cells. The most highly overexpressed genes were SYK, TACI, CD74, CD82, and CDC42EP5. Genes linked to G(2)/M and germinal center were down-regulated. Comparison of the gene expression profiles of NMZL and FL showed enriched expression of CHIT1, TGFB1, and TACI in NMZL, and BCL6, LMO2, and CD10 in FL. NMZL displayed increased expression of miR-221, miR-223, and let-7f, whereas FL strongly expressed miR-494. Our study identifies new candidate diagnostic molecules for NMZL and reveals survival pathways activated in NMZL.

B-cell lymphoma mutations: improving diagnostics and enabling targeted therapies.

B-cell lymphomas comprise an increasing number of clinicopathological entities whose characterization has historically been based mainly on histopathological features. In recent decades, the analysis of chromosomal aberrations as well as gene and miRNA expression profile studies have helped distinguish particular tumor types and also enabled the detection of a number of targets with therapeutic implications, such as those activated downstream of the B-cell receptor. Our ability to identify the mechanisms involved in B-cell lymphoma pathogenesis has been boosted recently through the use of Next Generation Sequencing techniques in the analysis of human cancer. This work summarizes the recent findings in the molecular pathogenesis of B-cell neoplasms with special focus on those clinically relevant somatic mutations with the potential to be explored as candidates for the development of new targeted therapies. Our work includes a comparison between the mutational indexes and ranges observed in B-cell lymphomas and also with other solid tumors and describes the most striking mutational data for the major B-cell neoplasms. This review describes a highly dynamic field that currently offers many opportunities for personalized therapy, although there is still much to be gained from the further molecular characterization of these clinicopathological entities.

Splenic marginal zone lymphoma: comprehensive analysis of gene expression and miRNA profiling.

Splenic marginal zone lymphoma is a small B-cell neoplasm whose molecular pathogenesis is still essentially unknown and whose differentiation from other small B-cell lymphomas is hampered by the lack of specific markers. We have analyzed the gene expression and miRNA profiles of 31 splenic marginal zone lymphoma cases. For comparison, 7 spleens with reactive lymphoid hyperplasia, 10 spleens infiltrated by chronic lymphocytic leukemia, 12 spleens with follicular lymphoma, 6 spleens infiltrated by mantle cell lymphoma and 15 lymph nodes infiltrated by nodal marginal zone lymphoma were included. The results were validated by qRT-PCR in an independent series including 77 paraffin-embedded splenic marginal zone lymphomas. The splenic marginal zone lymphoma miRNA signature had deregulated expression of 51 miRNAs. The most highly overexpressed miRNAs were miR-155, miR-21, miR-34a, miR-193b and miR-100, while the most repressed miRNAs were miR-377, miR-27b, miR-145, miR-376a and miR-424. MiRNAs located in 14q32-31 were underexpressed in splenic marginal zone lymphoma compared with reactive lymphoid tissues and other B-cell lymphomas. Finally, the gene expression data were integrated with the miRNA profile to identify functional relationships between genes and deregulated miRNAs. Our study reveals miRNAs that are deregulated in splenic marginal zone lymphoma and identifies new candidate diagnostic molecules for splenic marginal zone lymphoma.

Combinatorial effects of microRNAs to suppress the Myc oncogenic pathway.

Many mammalian transcripts contain target sites for multiple miRNAs, although it is not clear to what extent miRNAs may coordinately regulate single genes. We have mapped the interactions between down-regulated miRNAs and overexpressed target protein-coding genes in murine and human lymphomas. Myc, one of the hallmark oncogenes in these lymphomas, stands out as the up-regulated gene with the highest number of genetic interactions with down-regulated miRNAs in mouse lymphomas. The regulation of Myc by several of these miRNAs is confirmed by cellular and reporter assays. The same approach identifies MYC and multiple Myc targets as a preferential target of down-regulated miRNAs in human Burkitt lymphoma, a pathology characterized by translocated MYC oncogenes. These results indicate that several miRNAs must be coordinately down-regulated to enhance critical oncogenes, such as Myc. Some of these Myc-targeting miRNAs are repressed by Myc, suggesting that these tumors are a consequence of the unbalanced activity of Myc versus miRNAs.

miRNA expression in diffuse large B-cell lymphoma treated with chemoimmunotherapy.

Diffuse large B-cell lymphoma (DLBCL) prognostication requires additional biologic markers. miRNAs may constitute markers for cancer diagnosis, outcome, or therapy response. In the present study, we analyzed the miRNA expression profile in a retrospective multicenter series of 258 DLBCL patients uniformly treated with chemoimmunotherapy. Findings were correlated with overall survival (OS) and progression-free survival (PFS). miRNA and gene-expression profiles were studied using microarrays in an initial set of 36 cases. A selection of miRNAs associated with either DLBCL molecular subtypes (GCB/ABC) or clinical outcome were studied by multiplex RT-PCR in a test group of 240 cases with available formalin-fixed, paraffin-embedded (FFPE) diagnostic samples. The samples were divided into a training set (123 patients) and used to derive miRNA-based and combined (with IPI score) Cox regression models in an independent validation series (117 patients). Our model based on miRNA expression predicts OS and PFS and improves upon the predictions based on clinical variables. Combined models with IPI score identified a high-risk group of patients with a 2-year OS and a PFS probability of < 50%. In summary, a precise miRNA signature is associated with poor clinical outcome in chemoimmunotherapy-treated DLBCL patients. This information improves upon IPI-based predictions and identifies a subgroup of candidate patients for alternative therapeutic regimens.

Immediate effect of the use of toe separators on dynamic balance and ankle range of motion: a pilot study.

BACKGROUND AND OBJECTIVES: Injuries involving ankle stability and range of motion are among the most frequent in athletes and in the general population. In response, this study aimed to assess the immediate effects of toe separators on dynamic stability and ankle range of motion in healthy young individuals. METHODS: Among the 68 eligible participants, 50 healthy and active subjects completed all trials. The impact of the intervention was evaluated using the Weight Bearing Lunge Test and Y-Test. The control condition performed the tests without toe separators, while the experimental condition performed the tests with toe separators. All participants performed both conditions with a wash-out period of at least 7 days between trials. RESULTS: Statistical analysis revealed no significant differences in dynamic balance (p > 0.05) and range of motion (p > 0.05) between the two conditions. Additionally, no asymmetries were detected between the lower limbs in both tests (p > 0.05). CONCLUSIONS: The results of this pilot study indicate that using toe separators does not have an immediate effect on ankle range of motion and dynamic balance in young, healthy individuals. Future research should consider evaluating intervention programs of longer duration and exploring different populations.

Prevalence rates of chronic kidney disease and its association with cardiometabolic factors and cardiovascular diseases. SIMETAP-CKD study. / Tasas de prevalencia de enfermedad renal crónica y su asociación con factores cardiometabólicos y enfermedades cardiovasculares. Estudio SIMETAP-ERC.

INTRODUCTION: Chronic kidney disease (CKD) is a major health problem that contributes to the development of cardiovascular disorders such as heart failure and arteriosclerotic cardiovascular disease (ACVD). The aims of this study were to determine the prevalence of CKD and to assess its association with ACVD and cardiometabolic risk factors. METHODS: Cross-sectional observational study conducted in primary care setting. Population-based random sample: 6,588 people between 18 and 102 years old (response rate: 66%). Crude and sex- and age-adjusted prevalence rates of CKD according to KDIGO were determined by assessing albuminuria and estimated glomerular filtration rate according to CKD-EPI, and their associations with cardiometabolic factors and ACVD were determined. RESULTS: The crude prevalence of CKD was 11.48% (95%CI: 10.72-12.27%), without significant difference between men (11.64% [95%CI: 10.49-12.86%]) and women (11.35% [95%CI: 10.34-12.41%]). The age- and sex-adjusted prevalence rate of CKD was 9.16% (men: 8.61%; women: 9.69%). The prevalence of low estimated glomerular filtration rate (<60mL/min/1.73m2) and albuminuria (≥30mg/g) were 7.95% (95%CI: 7.30-8.61) and 5.98% (95%CI: 5.41-6.55), respectively. Hypertension, diabetes, prediabetes, increased waist-to-height ratio, heart failure, atrial fibrillation, and ACVD were independently associated with CKD (P<.001). Very high cardiovascular risk according to SCORE was found in 77.51% (95%CI: 74.54-80.49) of the population with CKD. CONCLUSIONS: The adjusted prevalence of CKD was 9.2% (low estimated glomerular filtration rate: 8.0%; albuminuria: 6.0%). Most of the patients with CKD had very high cardiovascular risk. Hypertension, diabetes, prediabetes, increased waist-to-height ratio and ACVD were independently associated with CKD.

Genetic Alterations in Members of the Proteasome 26S Subunit, AAA-ATPase (PSMC) Gene Family in the Light of Proteasome Inhibitor Resistance in Multiple Myeloma.

For the treatment of Multiple Myeloma, proteasome inhibitors are highly efficient and widely used, but resistance is a major obstacle to successful therapy. Several underlying mechanisms have been proposed but were only reported for a minority of resistant patients. The proteasome is a large and complex machinery. Here, we focus on the AAA ATPases of the 19S proteasome regulator (PSMC1-6) and their implication in PI resistance. As an example of cancer evolution and the acquisition of resistance, we conducted an in-depth analysis of an index patient by applying FISH, WES, and immunoglobulin-rearrangement sequencing in serial samples, starting from MGUS to newly diagnosed Multiple Myeloma to a PI-resistant relapse. The WES analysis uncovered an acquired PSMC2 Y429S mutation at the relapse after intensive bortezomib-containing therapy, which was functionally confirmed to mediate PI resistance. A meta-analysis comprising 1499 newly diagnosed and 447 progressed patients revealed a total of 36 SNVs over all six PSMC genes that were structurally accumulated in regulatory sites for activity such as the ADP/ATP binding pocket. Other alterations impact the interaction between different PSMC subunits or the intrinsic conformation of an individual subunit, consequently affecting the folding and function of the complex. Interestingly, several mutations were clustered in the central channel of the ATPase ring, where the unfolded substrates enter the 20S core. Our results indicate that PSMC SNVs play a role in PI resistance in MM.

Prevalence of prediabetes and association with cardiometabolic and renal factors. SIMETAP-PRED study. / Prevalencia de prediabetes y asociación con factores cardiometabólicos y renales. Estudio SIMETAP-PRED.

INTRODUCTION: Prediabetes is a major public health problem. The aims of the SIMETAP-PRED study were to determine the prevalence rates of prediabetes according to two diagnostic criteria, and to compare the association of cardiometabolic and renal risk factors between populations with and without prediabetes. METHODS: Cross-sectional observational study conducted in Primary Care. Based random sample: 6,588 study subjects (response rate: 66%). Two diagnostic criteria for prediabetes were used: 1) prediabetes according to the Spanish Diabetes Society (PRED-SDS): Fasting plasma glucose (FPG) 110-125mg/dL or HbA1c 6.0% -6.4%; 2) prediabetes according to the American Diabetes Association (PRED-ADA): FPG 100-125mg/dL or HbA1c 5.7%-6.4%. The crude and sex- and age-adjusted prevalence rates, and cardiometabolic and renal variables associated with prediabetes were assessed. RESULTS: The crude prevalence rates of PRED-SDS and PRED-ADA were 7.9% (95% CI 7.3-8.6%), and 22.0% (95% CI 21.0-23.0%) respectively, their age-adjusted prevalence rates were 6.6% and 19.1 respectively. The high or very high cardiovascular risk of the PRED-SDS or PRED-ADA populations were 68.6% (95%CI 64.5-72.6%) and 61.7% (95%CI 59.1-64.1%) respectively. Hypertension, hypertriglyceridemia, overweight, obesity, and increased waist-to-height ratio were independently associated with PRED-SDS. In addition to these factors, low glomerular filtration rate and hypercholesterolemia were also independently associated with PRED-ADA. CONCLUSIONS: The prevalence of PRED-ADA triples that of PRED-SDS. Two thirds of the population with prediabetes had a high cardiovascular risk. Several cardiometabolic and renal risk factors were associated with prediabetes. Compared to the SDS criteria, the ADA criteria make the diagnosis of prediabetes easier.

Prevalence rates of overweight and obesity and their associations with cardiometabolic and renal factors. SIMETAP-OB study. / Tasas de prevalencia de sobrepeso y obesidad y sus asociaciones con factores cardiometabólicos y renales. Estudio SIMETAP-OB.

INTRODUCTION: Excess weight is a major health problem. Aims of this study were to determine the prevalence rates of overweight and obesity, and to compare their associations with cardiometabolic and renal risk factors between obese and non-obese populations, and between overweight and non-overweight populations. METHODS: Cross-sectional observational study conducted in Primary Care. Population-based random sample: 6,588 study subjects between 18 and 102 years of age (response rate: 66%). Crude and sex- and age-adjusted prevalence rates of overweight and obesity were calculated, and their associations with cardiometabolic and renal variables were assessed by bivariate and multivariate analysis. RESULTS: The age- and sex-adjusted prevalence rates of overweight and obesity were 36.0% (42.1% in men; 33.1% in women) and 25.0% (26.2% in men; 24.5% in women), respectively. These prevalences increased with age, and were higher in men than in women. Fifty-two percent (95%CI: 50.0-53.9) of the overweight population and 62.3% (95%CI: 60.1-64.5) of the obese population had a high or very high cardiovascular risk. Abdominal obesity, physical inactivity, prediabetes, hypertension, hypertriglyceridemia, and low HDL-C were independently associated with both entities. Furthermore, diabetes was independently associated with overweight and hypercholesterolemia with obesity. CONCLUSIONS: The prevalence of overweight and obesity was 61.0% (68.4% in men and 59.0% in women). More than half of the overweight population and nearly two-thirds of the obese population had a high cardiovascular risk. Hyperglycemia, physical inactivity, hypertension, hypercholesterolemia, low HDL-C, and hypertriglyceridemia were independently associated with overweight and obesity.

Deregulated expression of the polycomb-group protein SUZ12 target genes characterizes mantle cell lymphoma.

Polycomb proteins are known to be of great importance in human cancer pathogenesis. SUZ12 is a component of the Polycomb PRC2 complex that, along with EZH2, is involved in embryonic stem cell differentiation. EZH2 plays an essential role in many cancer types, but an equivalent involvement of SUZ12 has not been as thoroughly demonstrated. Here we show that SUZ12 is anomalously expressed in human primary tumors, especially in mantle cell lymphoma (MCL), pulmonary carcinomas and melanoma, and is associated with gene locus amplification in some cases. Using MCL as a model, functional and genomic studies demonstrate that SUZ12 loss compromises cell viability, increases apoptosis, and targets genes involved in central oncogenic pathways associated with MCL pathogenesis. Our results support the hypothesis that the abnormal expression of SUZ12 accounts for some of the unexplained features of MCL, such as abnormal DNA repair and increased resistance to apoptosis.

Related cardiometabolic factors and prevalence of low HDL-cholesterol levels and atherogenic dyslipidemia. SIMETAP-AD study. / Factores cardiometabólicos asociados y prevalencia de concentraciones bajas de colesterol HDL y de dislipidemia aterogénica. Estudio SIMETAP-DA.

AIM: To determine the crude and sex- and age-adjusted prevalence rates of atherogenic dyslipidemia (AD) and low HDL-cholesterol levels (low-HDLc), and to assess their associations with cardiovascular risk factors, chronic kidney disease, cardiovascular and cardiometabolic diseases. METHODS: Population-based cross-sectional study conducted in Primary Care, with randomly selected adult subjects. The AD was considered if the patients had hypertriglyceridemia (triglycerides≥150mg/dL) and low-HDLc (<40mg/dL [men];<50mg/dL [women]). Crude and sex- and age-adjusted prevalence rates were determined, and univariate and multivariate analysis were performed to assess related cardiometabolic factors. RESULTS: Study population with 6,588 adults (55.9% women) with mean age 55.1 (±17.5) years. The mean HDLc levels were 49.2 (±12.6) mg/dL in men and 59.2 (±14.7) mg/dL in women. The crude prevalence rates of low-HDLc and AD were 30.8% (95%CI: 29.7-31.9), and 14.3% (95%CI: 13.5-15.2), respectively. The adjusted prevalence rates of low-HDLc were 28.0% in men and 31.0% in women, and AD were 16.4% in men and 10.6% in women. Seventy-three percent of the population with AD had high or very high cardiovascular risk. The independent factors associated with low HDLc or with AD were diabetes, smoking, abdominal obesity, and obesity. The major factors associated with low HDLc and AD were hypertriglyceridemia and diabetes, respectively. CONCLUSIONS: Almost a third of the adult population had low HDL-C and half of them met AD criteria. Cardiometabolic factors were associated with low HDL-C and AD, highlighting hypertriglyceridemia with low HDLc, and DM with AD.

Participation in Everyday Activities of Children with and without Neurodevelopmental Disorders: A Cross-Sectional Study in Spain.

Children with neurodevelopmental disorders (NDDs) often report significant difficulties performing activities of daily living (ADLs), which may restrict their daily participation. The aim of this study was to investigate the differences in ADLs participation between children with NDDs and typically developing (TD) children, and to explore the associations between different daily participation contexts. A cross-sectional study was conducted that included twenty children with a medical diagnosis of an NDD and 26 sex- and age-matched TD controls. The daily participation across home, community, school, and instrumental living activities was measured using the Child and Adolescent Scale of Participation (CASP). The results show that children with NDDs engaged in lower participation in all CASP contexts (Δ = 1.7-5.5, p < 0.001) and had a significantly higher prevalence of moderate or severe restricted participation than their TD peers (OR = 23.4, 95% CI = 3.6-154.2, p < 0.001). Additionally, a strong association was found between the different contexts of participation (r = 0.642-0.856). Overall, the children with NDDs experienced significant participation restrictions on their daily activities. This study adds to the growing evidence showing that intervention strategies in this population should adopt a participation-oriented approach.