Follicular lymphoma (FL) is the most frequent indolent lymphoma. Some patients (10%-15%) experience histologic transformation (HT) to a more aggressive lymphoma, usually diffuse large B-cell lymphoma ...(DLBCL). This study aimed to validate and improve a genetic risk model to predict HT at diagnosis.We collected mutational data from diagnosis biopsies of 64 FL patients. We combined them with the data from a previously published cohort (total n = 104; 62 from nontransformed and 42 from patients who did transform to DLBCL). This combined cohort was used to develop a nomogram to estimate the risk of HT. Prognostic mutated genes and clinical variables were assessed using Cox regression analysis to generate a risk model. The model was internally validated by bootstrapping and externally validated in an independent cohort. Its performance was evaluated using a concordance index and a calibration curve.
The clinicogenetic nomogram included the mutational status of 3 genes (HIST1HE1, KMT2D, and TNFSR14) and high-risk Follicular Lymphoma International Prognostic Index and predicted HT with a concordance index of 0.746. Patients were classified as being at low or high risk of transformation. The probability HT function at 24 months was 0.90 in the low-risk group vs 0.51 in the high-risk group and, at 60 months, 0.71 vs 0.15, respectively. In the external validation cohort, the probability HT function in the low-risk group was 0.86 vs 0.54 in the high-risk group at 24 months, and 0.71 vs 0.32 at 60 months. The concordance index in the external cohort was 0.552. In conclusion, we propose a clinicogenetic risk model to predict FL HT to DLBLC, combining genetic alterations in HIST1H1E, KMT2D, and TNFRSF14 genes and clinical features (Follicular Lymphoma International Prognostic Index) at diagnosis. This model could improve the management of FL patients and allow treatment strategies that would prevent or delay transformation.
A burial and a rich offering were found under Room 2 in the Murals Building, Bonampak, a Mayan archaeological site situated in Chiapas, Mexico. This burial may be related with the creation of the ...famous mural paintings. A rich set of jewelry made of green stones was among the different objects found. Green stones have great importance in Mesoamerican cultures, those composed of jadeite being the most appreciated. To characterize the green stones, different spectroscopic techniques were used in a complementary way: Raman and infrared spectroscopies (FT-IR) were used for global mineralogical analysis, while X-ray diffraction (XRD) and X-ray fluorescence (XRF) were applied simultaneously in situ on the artifacts that were not successfully identified by these molecular techniques. In addition, XRF was used to contrasts the elemental information from pieces composed of pyroxenes that may be related to the raw sources of jade in Guatemala. The main minerals identified within the beads and earrings were jadeite with omphacite and jadeite with albite; to a minor extent, quartz, and serpentine. In this paper, the main features of the molecular and X-ray techniques are compared in order to determine the advantages and limitations of these spectroscopies for mineral identification. With this combination of techniques, it was possible to undertake a suitable characterization of the analyzed objects. This paper focuses on the XRD–XRF combined analysis for in situ noninvasive characterization.
The overlap of morphology and immunophenotype between angioimmunoblastic T-cell lymphoma (AITL) and other nodal peripheral T-cell lymphomas (n-PTCLs) is a matter of current interest whose clinical ...relevance and pathogenic background have not been fully established. We studied a series of 98 n-PTCL samples (comprising 57 AITL and 41 PTCL-NOS) with five T
antibodies (CD10, BCL-6, PD-1, CXCL13, ICOS), looked for mutations in five of the genes most frequently mutated in AITL (
,
and
) using the Next-Generation-Sequencing Ion Torrent platform, and measured the correlations of these characteristics with morphology and clinical features. The percentage of mutations in the
and
genes was similar (23.5% of cases).
was mutated in 14.3%,
in 11.2% and
in 7.1% of cases, respectively. In the complete series, mutations in
gene were associated with the presence of mutations in
and
(
0.001,
0.043, and
0.029, respectively). Fourteen cases featured
mutations without
mutations. A close relationship was found between the presence of these mutations and a T
-phenotype in AITL and PTCL-NOS patients. Interestingly, BCL-6 expression was the only T
marker differentially expressed between AITL and PTCL-NOS cases. There were many fewer mutated cases than there were cases with a T
phenotype. Overall, these data suggest alternative ways by which neoplastic T-cells overexpress these proteins. On the other hand, no clinical or survival differences were found between any of the recognized subgroups of patients with respect to their immunohistochemistry or mutational profile.
Diffuse large B-cell lymphoma (DLBCL) is an aggressive and heterogeneous disease with variable prognosis associated with clinical features, cell-of-origin and genetic aberrations. The main problems ...for DLBCL patients are that a substantial percentage of them (30-40%) are refractory to treatment or relapse and the lack of accurate predictive markers that adequately determine which patients will benefit from immunochemotherapy. Several recent deep-sequencing studies have proposed new genetic subtypes based on the DLBCL genomic profile (Wright et al., 2020; Lacy et al. 2020) and associated with clinical outcome. However, a consensus and validated classification is still needed. The aim was to determine whether genetic alterations of individual genes or genes clustered in pathways are associated with clinical outcome in immunochemotherapy-treated patients.
We used targeted massive sequencing to analyze 84 diagnostic samples from a multicentric cohort of patients with DLBCL treated with rituximab-containing therapies with a median follow up of 6 years. A two-step genetic classifier was built with mutation information from 27 genes and BCL2/BCL6 translocations, based on recently proposed genetic subtypes (Wright et al., 2020; Lacy et al. 2020). Logistic regression and Kaplan-Meier analyses for survival and risk of relapse were performed to assess the potential clinical value of the classifier. Moreover, this two-step classifier was validated in an external cohort and its specificity and sensitivity were tested to determine its accuracy in classifying patients compared to the previous approaches.
We found that the most frequently mutated genes were IGLL5 (43%), KMT2D (33.3%), CREBBP (28.6%), PIM1 (26.2%), and CARD11 (22.6%). Mutations in CD79B, PRDM1, and NOTCH2 were associated with a higher risk of relapse after treatment, whereas patients with mutations in CD79B, ETS1, PRDM1, and TNFAIP3 had significantly shorter survival. Analyzing the impact of gene mutations on predefined gene sets related to lymphomagenesis revealed that mutations in genes involved in B-cell development, and BCR-PI3K and MAPK-ERK pathways were significantly associated with a higher risk of relapse and/or shorter overall survival. These results confirmed the current landscape of genetic alterations in DLBCL.
According to the two-step classifier, we categorized the samples into MCD, BN2, EZB, ST2, and N1 genetic subgroups (Figure). We tested the accuracy of our classification in an external series (UK population-based Haematological Malignancy Research Network cohort HMRN) to determine its specificity and sensitivity compared with their own classification (Lacy et al. 2020) and the LymphGen algorithm (Wright et al., 2020). The comparison demonstrated (Wright et al., 2020; Lacy et al. 2020), a specificity and sensitivity higher than 85% for each subtype, except for BN2. This controversy may be explained by the fact that BN2 is less strongly defined than the other subtypes.
We then tested their clinical impact and found the EZB and ST2 subtypes to have a better clinical course and a lower risk of relapse. The BN2 group had the worst clinical outcome of our series, unlike other published studies (Figure). These results were validated in the HMRN cohort, in which N1 showed a higher risk of relapse and shorter overall survival, whereas ST2 is the group with the most favorable outcome. Although N1 was not included for clinical outcome analysis in our cohort due to the small number of cases, the validation of our classifier defined N1 as the most aggressive subtype.
In summary, we propose and validate a feasible genetic DLBCL classifier based on an optimized panel of genes that unifies previous genetic classification algorithms in such a way as to facilitate its implementation as part of pathology laboratories for routine patient management. This genetic classifier, combined with clinical data and other molecular characteristics, should eventually help develop improved risk models for DLBCL patients, and guide precision therapy.
Display omitted
Perez Callejo:F. Hoffmann-La Roche: Current Employment, Current equity holder in publicly-traded company.
Follicular lymphoma (FL) is the most frequent indolent lymphoma. 10-15% of patients suffer histological transformation (HT) to a more aggressive lymphoma, usually diffuse large B cell lymphoma ...(DLBCL). This study aimed to validate and improve a genetic risk model to predict HT at diagnosis. We collected mutational data from diagnosis biopsies of 64 FL patients. We combined them with the data from a previously published cohort (total n = 104, 62 from non-transformed, and 42 from patients who did transform to DLBCL). This combined cohort was used to develop a nomogram to estimate the risk of HT. Prognostic mutated genes and clinical variables were assessed using Cox regression analysis to generate a risk model. The model was internally validated by bootstrapping and externally validated in an independent cohort. Its performance was evaluated using a concordance index and a calibration curve. The clinicogenetic nomogram included the mutational status of three genes (HIST1HE1, KMT2D, and TNFSR14) and high-risk FLIPI and predicted HT with a concordance index of 0.746. Patients were classified as being at low or high risk of transformation. The probability HT function at 24 months was 0.90 in the low-risk group vs. 0.51 in the high-risk group and, at 60 months, 0.69 vs. 0.15, respectively. In the external validation cohort, the probability HT function in the low-risk group was 0.86 vs. 0.54 in the high-risk group at 24 months, and 0.71 vs. 0.32 at 60 months. The concordance index in the external cohort was 0.552. In conclusion, we propose a clinicogenetic risk model to predict FL HT to DLBLC, combining genetic alterations in HIST1H1E, KMT2D, and TNFRSF14 genes and clinical features (FLIPI) at diagnosis. This model could improve the management of FL patients and allow treatment strategies that would prevent or delay transformation.
INTRODUCTION
Development of targeted therapy in Cutaneous T-Cell Lymphoma (CTCL) patients still requires actionable mutated genes and deregulated pathways to be identified. We have recently published ...the mutational status of a number of human CTCL lesions, and found JAK/STAT signaling pathway to frequently harbor somatic mutations (Vaqué et al 2014). In this line of evidence, activating mutations in JAK kinases have been reported in human hematological malignancies (Kameda T1 2010) and may serve as indicators for targeted therapy. Therefore, we decided to analyze the mutational status of JAK/STAT pathway in a greater cohort of human CTCL patient samples and cell lines, by using massive parallel sequencing techniques, to shed light on its possible role in the pathogenesis and to uncover its potential as a new therapeutic target for this disease. To this end, we have studied the biological and molecular effects that targeted inhibition of the JAK/STAT signaling pathway, exerts in human CTCL cells, and explore its potential use as a targeted therapy.
MATERIAL AND METHODS
NGS: We searched for mutations in the catalytic domain of JAK and STAT genes in 39 CTCL patients using targeted NGS techniques: HaloPlex (Agilent) and sequencing in IonTorrent (LifeTech) and Illumina sequencers. Proliferation, apoptosis, cell cycle and DNA synthesis assays: CTCL cell lines were: My-La (MF), HH (MF) and HUT-78 (SS) (ECACC,Salisbury, UK). Effects in proliferation was assessed using CellTiter-Glo® Luminescent Cell Viability Assay (Promega, USA). To analyze the effects on cell survival, we evaluated early and delayed cell death provoked by FACS using FlowCellect Annexin Red Kit (EMD Millipore Corporation, USA). The distribution of cells among different phases of the cell cycle was evaluated by FACS using propidium iodide (PI, Sigma-Aldrich, USA) and Click-iT® EdU Alexa Fluor® 488 Flow Cytometry Assay Kit (Technologies-Thermo Fisher Scientific, USA) according to manufacturer’s instruction. Gene expression (GEP) studies: mRNA was extracted with Trizol and mRNA Array-based expression analysis was perfomed using a Whole Human Genome Agilent 4 × 44K v1 Oligonucleotide Microarray.
RESULTS:
Using our approach we found JAK/STAT pathway mutated in up to 24% of the CTCL samples. 3 mutations were found in JAK1, 5 in JAK3 and 1 in STAT5A genes. Interestingly, most of these mutations affected the tyrosine kinase domain of JAK kinases, a hotspot for activating mutations described in multiple types of human cancer.
Thereafter, we decided to explore the biological effects of targeted JAK/STAT inhibition, using specific JAK inhibitors (JAKi) in preclinical CTCL models. To this end, CTCL cell lines were incubated at different time points with increasing concentrations of JAKi. These treatments inhibited JAK/STAT signaling in CTCL cells, as assessed by western-blot using P-STAT-1,-3 and -5 antibodies. Biologically, JAKi provoked a marked inhibition of cell proliferation by a mechanism impinging the control of DNA replication. This was accompanied with a modest increase in cell death. To better understand the molecular mechanisms controlled by aberrant JAK/STAT signaling, we also performed an array-based mRNA expression analysis (GEP) in HUT78 cell line (JAK1Y654F) treated at different time points (0, 0.5 and 3h) with JAKi (at a concentration of IC50). Our results showed a number of genes regulated by JAK/STAT signaling which activity has been described as T-cell activation, differentiation and proliferation (i.e. PAG1,TNF or EGR1).These genes may serve as potential indicators for therapy using JAK/STAT inhibitors or as surrogate markers for drug response.
CONCLUSIONS
Our results show frequent mutations affecting JAK/STAT downstream signaling in samples from patients with CTCL. Targeted inhibition of this pathway reveals an aberrant CTCL growing controlled by this signaling. Thus, these results provide evidence for the use of JAK/STAT inhibitors on the basis of a targeted massive sequencing analysis in specific cases of CTCL.
No relevant conflicts of interest to declare.
Sulfur is an essential element in determining the productivity and quality of agricultural products. It is also an element associated with tolerance to biotic and abiotic stress in plants. In ...agricultural practice, sulfur has broad use in the form of sulfate fertilizers and, to a lesser extent, as sulfite biostimulants. When used in the form of bulk elemental sulfur, or micro- or nano-sulfur, applied both to the soil and to the canopy, the element undergoes a series of changes in its oxidation state, produced by various intermediaries that apparently act as biostimulants and promoters of stress tolerance. The final result is sulfate S
, which is the source of sulfur that all soil organisms assimilate and that plants absorb by their root cells. The changes in the oxidation states of sulfur S
to S
depend on the action of specific groups of edaphic bacteria. In plant cells, S
sulfate is reduced to S
and incorporated into biological molecules. S
is also absorbed by stomata from H
S, COS, and other atmospheric sources. S
is the precursor of inorganic polysulfides, organic polysulfanes, and H
S, the action of which has been described in cell signaling and biostimulation in plants. S
is also the basis of essential biological molecules in signaling, metabolism, and stress tolerance, such as reactive sulfur species (RSS), SAM, glutathione, and phytochelatins. The present review describes the dynamics of sulfur in soil and plants, considering elemental sulfur as the starting point, and, as a final point, the sulfur accumulated as S
in biological structures. The factors that modify the behavior of the different components of the sulfur cycle in the soil-plant-atmosphere system, and how these influences the productivity, quality, and stress tolerance of crops, are described. The internal and external factors that influence the cellular production of S
and polysulfides vs. other S species are also described. The impact of elemental sulfur is compared with that of sulfates, in the context of proper soil management. The conclusion is that the use of elemental sulfur is recommended over that of sulfates, since it is beneficial for the soil microbiome, for productivity and nutritional quality of crops, and also allows the increased tolerance of plants to environmental stresses.
PDF
