Molecular Insights into Major Peripheral T-cell Lymphoma Entities with Advances in a Representative Model System

Peripheral T-cell lymphoma (PTCL) is a group of complex clinicopathological entities associated with an aggressive clinical course. Angioimmunoblastic T-cell lymphoma (AITL) and PTCL-not otherwise specified (PTCL-NOS) are the two most frequent categories accounting for more than 50% of PTCLs. Gene expression profiling (GEP) defined molecular signatures for AITL and delineated biological and prognostic subgroups within PTCL-NOS (PTCL-GATA3 and PTCL-TBX21). Genomic copy number analysis and targeted sequencing revealed unique genomic abnormalities and oncogenic pathways, indicating distinct oncogenic evolution. PTCL-GATA3 exhibited higher genomic complexity characterized by frequent loss or mutation of tumor suppressor genes targeting the CDKN2A/B-TP53 axis and PTEN-PI3K pathways. PTCL-TBX21 had fewer abnormalities, primarily targeting cytotoxic effector genes, and was enriched in mutations of genes regulating DNA methylation. AITL showed lower genomic complexity compared to other PTCL entities, suggesting it is epigenetically driven, yet frequent co-occurring gains of chromosome 5 and 21 were observed. Thus, to further explore the underlying pathobiology of AITL, we established an AITL-like murine model. We have found TET2 to be the most recurrent mutation in AITL with evidence that it may be a founder mutation. The generated murine model had a loss of Tet2 in CD4+ T-cells, resulting in the long-term development of an aggressive T-cell lymphoma with a TFH phenotype, similar to AITL. These mice developed lymphomas characterized by massive splenomegaly, marked generalized lymphadenopathy, hepatomegaly and occasional involvement of other parenchymal organs. Serial RNA-sequencing analysis of the pre-neoplastic and neoplastic CD4+ T-cells revealed increased PI3K-AKT signaling and decreased Foxo1 signaling likely mediated through ICOS in the neoplastic cells, promoting TFH cell expansion. Overall, we demonstrated that novel GEP-defined PTCL subgroups likely evolve by distinct genetic pathways and that AITL is driven more by epigenetic alterations, such as loss of Tet2 as it resulted in an aggressive T-cell lymphoma with a TFH phenotype. These findings emphasize the critical need for genetically faithful model systems for studying these common PTCL entities in order to improve therapeutic options

Crying with laughter: adapting the tickling protocol to address individual differences among rats in their response to playful handling

International audienc

Caveolin-1 is dispensable for early lymphoid development, but plays a role in the maintenance of the mature splenic microenvironment

Objective Caveolin-1 (CAV1) is known for its role as both a tumor suppressor and an oncogene, harboring a highly context-dependent role within a myriad of malignancies and cell types. In an immunological context, dysregulation of CAV1 expression has been shown to alter immunological signaling functions and suggests a pivotal role for CAV1 in the facilitation of proper immune responses. Nonetheless, it is still unknown how Cav1-deficiency and heterozygosity would impact the development and composition of lymphoid organs in mice. Herein, we investigated the impacts of Cav1-dysregulation on the lymphoid organs in young (12 weeks) and aged (36 weeks) Cav1+/+, Cav1+/−, and Cav1−/− mice. Results We observed that only Cav1-deficiency is associated with persistent splenomegaly at all timepoints. Furthermore, no differences in overall body weight were detected (and without sexual dimorphisms). Both aged Cav1+/− and Cav1−/− mice present with decreased CD19+CD22+ B cells and secondary-follicle atrophy, specifically in the spleen, compared with wild-type controls and irrespective of splenomegaly status. Consequently, the demonstrated effects on B cell homeostasis and secondary follicle characteristics prompted our investigation into follicle-derived human B-cell lymphomas. Our investigation points toward CAV1 as a dysregulated protein in follicle-derived B-cell malignancies without harboring a differential expression between more aggressive and indolent hematological malignancies

Methodological checklists for improving research quality and reporting consistency

This is the author accepted manuscript. The final version is available from Cambridge University Press via the DOI in this recor

Evaluating the Effect of Chemical Digestion Treatments on Polystyrene Microplastics: Recommended Updates to Chemical Digestion Protocols

Establishing the toxicity and exposure consequences of microplastics (MPs) on marine organisms relies on the nondestructive isolation of plastics from biological matrices. MPs are commonly extracted from these matrices by chemical digestion using alkali (e.g., potassium hydroxide (KOH) and sodium hydroxide (NaOH)), oxidative (e.g., hydrogen peroxide (H2O2)) and/or acidic (e.g., nitric acid (HNO3)) reagents. Although these digestion conditions can be highly effective for MP extraction, they can also react with the plastics. This can attribute an inaccurate representation of plastic contamination by altering MP visual characteristics (size, shape, color), thereby impeding identification and potentially returning erroneous numbers of ingested particles. In this study, the degradative impacts are assessed of the routinely applied digestion reagents (i) KOH, (ii) NaOH, (iii) H2O2, and (iv)HNO3 on polystyrene (PS) based MPs sized between 200 μm and 5 mm. Degradation of the PS MPs is evaluated using FT-IR, gel permeation chromatography, NMR, photoluminescence spectroscopy, and microscopy. These studies reveal HNO3 to be the most destructive for PS MPs, while the alkali and oxidative reagents result in negligible changes in plastic properties. These results are recommended to be used as a guideline to update current protocols to ensure the nondestructive treatment of MPs

Human Umbilical Cord Therapy Improves Long-Term Behavioral Outcomes Following Neonatal Hypoxic Ischemic Brain Injury

Background: Hypoxic ischemic (HI) insult in term babies at labor or birth can cause long-term neurodevelopmental disorders, including cerebral palsy (CP). The current standard treatment for term infants with hypoxic ischemic encephalopathy (HIE) is hypothermia. Because hypothermia is only partially effective, novel therapies are required to improve outcomes further. Human umbilical cord blood cells (UCB) are a rich source of stem and progenitor cells making them a potential treatment for neonatal HI brain injury. Recent clinical trials have shown that UCB therapy is a safe and efficacious treatment for confirmed cerebral palsy. In this study, we assessed whether early administration of UCB to the neonate could improve long-term behavioral outcomes and promote brain repair following neonatal HI brain injury.Methods: HI brain injury was induced in postnatal day (PND) 7 rat pups via permanent ligation of the left carotid artery, followed by a 90 min hypoxic challenge. UCB was administered intraperitoneally on PND 8. Behavioral tests, including negative geotaxis, forelimb preference and open field test, were performed on PND 14, 30, and 50, following brains were collected for assessment of neuropathology.Results: Neonatal HI resulted in decreased brain weight, cerebral tissue loss and apoptosis in the somatosensory cortex, as well as compromised behavioral outcomes. UCB administration following HI improved short and long-term behavioral outcomes but did not reduce long-term histological evidence of brain injury compared to HI alone. In addition, UCB following HI increased microglia activation in the somatosensory cortex compared to HI alone.Conclusion: Administration of a single dose of UCB cells 24 h after HI injury improves behavior, however, a single dose of cells does not modulate pathological evidence of long-term brain injury

Characterization of Antibodies against Receptor Activity-Modifying Protein 1 (RAMP1): A Cautionary Tale

Calcitonin gene-related peptide (CGRP) is a key component of migraine pathophysiology, yielding effective migraine therapeutics. CGRP receptors contain a core accessory protein subunit: receptor activity-modifying protein 1 (RAMP1). Understanding of RAMP1 expression is incomplete, partly due to the challenges in identifying specific and validated antibody tools. We profiled antibodies for immunodetection of RAMP1 using Western blotting, immunocytochemistry and immunohistochemistry, including using RAMP1 knockout mouse tissue. Most antibodies could detect RAMP1 in Western blotting and immunocytochemistry using transfected cells. Two antibodies (844, ab256575) could detect a RAMP1-like band in Western blots of rodent brain but not RAMP1 knockout mice. However, cross-reactivity with other proteins was evident for all antibodies. This cross-reactivity prevented clear conclusions about RAMP1 anatomical localization, as each antibody detected a distinct pattern of immunoreactivity in rodent brain. We cannot confidently attribute immunoreactivity produced by RAMP1 antibodies (including 844) to the presence of RAMP1 protein in immunohistochemical applications in brain tissue. RAMP1 expression in brain and other tissues therefore needs to be revisited using RAMP1 antibodies that have been comprehensively validated using multiple strategies to establish multiple lines of convincing evidence. As RAMP1 is important for other GPCR/ligand pairings, our results have broader significance beyond the CGRP field

Successful recruitment to trials : findings from the SCIMITAR+ Trial

BACKGROUND: Randomised controlled trials (RCT) can struggle to recruit to target on time. This is especially the case with hard to reach populations such as those with severe mental ill health. The SCIMITAR+ trial, a trial of a bespoke smoking cessation intervention for people with severe mental ill health achieved their recruitment ahead of time and target. This article reports strategies that helped us to achieve this with the aim of aiding others recruiting from similar populations. METHODS: SCIMITAR+ is a multi-centre pragmatic two-arm parallel-group RCT, which aimed to recruit 400 participants with severe mental ill health who smoke and would like to cut down or quit. The study recruited primarily in secondary care through community mental health teams and psychiatrists with a smaller number of participants recruited through primary care. Recruitment opened in October 2015 and closed in December 2016, by which point 526 participants had been recruited. We gathered information from recruiting sites on strategies which led to the successful recruitment in SCIMITAR+ and in this article present our approach to trial management along with the strategies employed by the recruiting sites. RESULTS: Alongside having a dedicated trial manager and trial management team, we identified three main themes that led to successful recruitment. These were: clinicians with a positive attitude to research; researchers and clinicians working together; and the use of NHS targets. The overriding theme was the importance of relationships between both the researchers and the recruiting clinicians and the recruiting clinicians and the participants. CONCLUSIONS: This study makes a significant contribution to the limited evidence base of real-world cases of successful recruitment to RCTs and offers practical guidance to those planning and conducting trials. Building positive relationships between clinicians, researchers and participants is crucial to successful recruitment

The Genetic Basis of Hepatosplenic T-cell Lymphoma

Hepatosplenic T cell lymphoma (HSTL) is a rare and lethal lymphoma; the genetic drivers of this disease are unknown. Through whole exome sequencing of 68 HSTLs, we define recurrently mutated driver genes and copy number alterations in the disease. Chromatin modifying genes including SETD2, INO80 and ARID1B were commonly mutated in HSTL, affecting 62% of cases. HSTLs manifest frequent mutations in STAT5B (31%), STAT3 (9%), and PIK3CD (9%) for which there currently exist potential targeted therapies. In addition, we noted less frequent events in EZH2, KRAS and TP53. SETD2 was the most frequently silenced gene in HSTL. We experimentally demonstrated that SETD2 acts as a tumor suppressor gene. In addition, we found that mutations in STAT5B and PIK3CD activate critical signaling pathways important to cell survival in HSTL. Our work thus defines the genetic landscape of HSTL and implicates novel gene mutations linked to HSTL pathogenesis and potential treatment targets

Feel-good songs: application of a novel playback paradigm to induce a positive affective state in juvenile male Wistar rats

Across animal welfare science there is a lack of validated models of positive affective states. Previous work has shown that presentation of contrastingly valenced ultrasonic vocalisations (USVs) to rats alters their behaviour. However, the potential of using playback of USVs to induce a positive affective state and promote positive animal welfare has yet to be explored. We used three cohorts of juvenile male Wistar rats (37 days old) in three independent experiments to develop a novel home cage playback paradigm to induce a positive affective state in rats. The intention behind this paradigm was to create a low-stress environment, given the heightened susceptibility of positive affective states to stress. Rats were presented in pairs with a playback track consisting of positively valenced 50-kHz USVs, White Noise (within the 30 – 100kHz range), or Background Noise in their home cage. In Experiments 1 (N = 7 cages) and 2 (N = 14 cages), rats received a single presentation of each playback track in a Latin square experimental design. In Experiment 3 (N = 20 cages), rats received repeated presentations of the same playback track over five consecutive days. Changes in affective state were measured through USV production, approach to the stimulus, and play behaviour. Across all three experiments, the presentation of 50-kHz stimuli USVs increased subject-produced positively valenced 50kHz USVs compared to presentation of Background Noise (e.g. Experiment 2; F2,239 = 6.05, p < 0.05). Similarly, rats also expressed an increase in approach behaviour towards the speaker in response to 50kHz stimuli USVs compared to White Noise and Background Noise (Experiment 3 duration of approach behaviour; F2,479 = 10.55, p < 0.001). Whilst there was complexity in the relationship between the presentation of different acoustic stimuli and play behaviour, rats presented with the 50kHz stimuli showed increased social play in the ten minutes during presentation under some of our test conditions. The impact of acoustic stimuli on measures of affective state across cohorts provides evidence that the home cage playback paradigm holds promise as a method for inducing a positive affective state in rats