Citations

Abstract Introduction: Idiopathic multicentric Castleman disease (iMCD) is a rare lymphoproliferative disorder characterized by systemic inflammation of unknown etiology. Clinically, iMCD is classified into idiopathic plasmacytic lymphadenopathy (IPL), thrombocytopenia, anasarca, fever, renal dysfunction/reticulin fibrosis, and organomegaly (TAFRO), and not otherwise specified (NOS) subtypes. Among these, TAFRO and a subset of NOS cases share overlapping clinicopathological features and often follow a fatal clinical course. However, the pathogenesis remains largely undefined. This study combines a comprehensive transcriptomic analysis with histological assessment, focusing on vascular proliferation to elucidate the molecular mechanisms of the TAFRO subtype. Methods: We conducted a pathological analysis of nine IPL, nine TAFRO, and eight NOS cases, and performed transcriptome analysis from frozen lymph node specimens from 12, two, and four IPL, TAFRO, and NOS cases, respectively. Differentially expressed genes (DEGs) and KEGG pathway analysis were performed to identify significant pathways in TAFRO and NOS, with IPL cases as controls. Results: Histologically, TAFRO and NOS exhibited a significantly higher degree of vascularization than IPL (IPL vs TAFRO: p < 0.001, IPL vs NOS: p = 0.002). In addition, “whirlpool vessels” in GCs were seen in most cases in TAFRO and NOS (TAFRO: 9/9, 100%, NOS: 6/8, 75%), but not in IPL (IPL vs TAFRO: p < 0.001, IPL vs NOS: p = 0.007). Immunostaining for Ets-related gene revealed that the number of endothelial cells of TAFRO and NOS was significantly larger than that in IPL (TAFRO vs IPL: p < 0.001, NOS vs IPL: p = 0.002). KEGG pathway analysis revealed the PI3K–Akt signaling pathway was significantly enriched in the TAFRO/NOS groups. Immunohistochemistry of phosphorylated Akt and phosphorylated p38, both upstream and downstream effectors of the PI3K–Akt signaling pathway, were positive in vascular endothelial cells of TAFRO and NOS. Discussion: TAFRO and NOS exhibit marked angiogenesis and are histologically distinct from IPL. Furthermore, this angiogenesis is thought to be driven by upregulation of the PI3K–Akt signaling pathway, which may also contribute to extravascular fluid leakage, potentially leading to pleural and ascitic effusion or systemic edema observed in the TAFRO subtype. Conclusion: The present results suggest the crucial role of PI3K–Akt signaling pathway in TAFRO pathogenesis from transcriptomic and histopathological findings. This mechanistic insight may help elucidate the distinct pathogenesis of iMCD subtypes.

Summary CD19 chimeric antigen receptor T‐cell (CAR‐T) therapy is a pivotal part of the treatment algorithm for large B‐cell lymphoma (LBCL) in the second‐ and third‐line settings based on numerous clinical trials. However, these studies included very few patients with a diagnosis of high‐grade B‐cell lymphoma, not otherwise specified (HGBCL‐NOS) and it is unclear if outcomes are similar to those observed with more common LBCL histologies. Using the Center for International Blood and Marrow Transplant Research (CIBMTR) registry, we identified 111 HGBCL‐NOS patients who received CAR‐T therapy. The cytokine release syndrome (CRS) rate was 74% (7.7% grade 3+), median onset was 4 days (1–17) and immune effector cell‐associated neurotoxicity syndrome rate was 45.2% (22.6% grade 3+), median onset was 7 days (1–17). At 2 years post‐CAR‐T infusion, the probability of overall survival and progression‐free survival were 41.6% and 28.7% respectively. The 2‐year non‐relapse mortality and relapse/progression were 3.1% (95% confidence interval [CI]: 0.6–7.6) and 68.1% (95% CI: 57.9–77.6) respectively. Our analysis illustrates that patients with HGBCL‐NOS represent a particularly difficult group to treat, even with CAR‐T therapy. We observed that one third of patients achieved a durable response; however, the overall results were less favourable, with lower overall response rate, overall survival and progression‐free survival as compared to published reports on LBCL.

RESUMO Introdução: estima-se que aproximadamente 20 milhões de pessoas sejam submetidas a cirurgia de hérnia inguinal anualmente no mundo, sendo a técnica de Lichtenstein o procedimento cirúrgico mais realizado. O objetivo desse estudo é analisar o conhecimento dos principios técnicos empregados na técnica de Lichtenstein. Método: estudo tipo levantamento interseccional aprovado pelo comitê de ética em pesquisa do Centro Universitário São Camilo (CAAE: 70036523.1.0000.0062). Durante o período da pesquisa foram encaminhados 11.622 e-mails aos membros das principais sociedades cirúrgicas nacionais com uma pesquisa sobre os princípios técnicos da cirurgia de Lichtenstein. A pesquisa foi realizada por formulário eletrônico com 10 questões de múltipla escolha. O formulário foi respondido de forma anônima nas plataformas SurveyMonkey e Google Forms. Resultado: foram recebidos 744 respostas ao formulário eletrônico. Com base nesse número de respondedores, nossa pesquisa apresenta grau de confiança de 95% com margem de erro de 3,5%. Foi observado que não há padronização da técnica entre a maioria dos respondedores (53.4%). Muitos cirurgiões ainda fazem dissecção digital do funículo espermático (47%). Um pequeno número de entrevistados (15,2%) realizam sutura com fio absorvível na região da aponeurose do obliquo interno, enquanto, mais da metade (55,2%) continua fazendo sutura com fio inabsorvível. A maior parte dos cirurgiões utilizam overlap pequeno ou fixam a tela justaposta a sínfise púbica (51%). Conclusão: nossa pesquisa identificou que uma porcentagem pequena dos entrevistados conhecem adequadamente os princípios técnicos da cirurgia de Lichtenstein. O resultado nos traz novas percepções sob a necessidade de revistar a consagrada técnica de Lichtenstein.

Abstract Peripheral T-cell lymphomas (PTCLs) are heterogenous T-cell neoplasms often associated with epigenetic dysregulation. We investigated de novo DNA methyltransferase 3A (DNMT3A) mutations in common PTCL entities, including angioimmunoblastic T-cell lymphoma and novel molecular subtypes identified within PTCL–not otherwise specified (PTCL-NOS) designated as PTCL-GATA3 and PTCL-TBX21. DNMT3A-mutated PTCL-TBX21 cases showed inferior overall survival (OS), with DNMT3A-mutated residues skewed toward the methyltransferase domain and dimerization motif (S881–R887). Transcriptional profiling demonstrated significant enrichment of activated CD8+ T-cell cytotoxic gene signatures in the DNMT3A-mutant PTCL-TBX21 cases, which was further validated using immunohistochemistry. Genomewide methylation analysis of DNMT3A-mutant vs wild-type (WT) PTCL-TBX21 cases demonstrated hypomethylation in target genes regulating interferon-γ (IFN-γ), T-cell receptor signaling, and EOMES (eomesodermin), a master transcriptional regulator of cytotoxic effector cells. Similar findings were observed in a murine model of PTCL with Dnmt3a loss (in vivo) and further validated in vitro by ectopic expression of DNMT3A mutants (DNMT3A-R882, -Q886, and -V716, vs WT) in CD8+ T-cell line, resulting in T-cell activation and EOMES upregulation. Furthermore, stable, ectopic expression of the DNMT3A mutants in primary CD3+ T-cell cultures resulted in the preferential outgrowth of CD8+ T cells with DNMT3AR882H mutation. Single-cell RNA sequencing(RNA-seq) analysis of CD3+ T cells revealed differential CD8+ T-cell subset polarization, mirroring findings in DNMT3A-mutated PTCL-TBX21 and validating the cytotoxic and T-cell memory transcriptional programs associated with the DNMT3AR882H mutation. Our findings indicate that DNMT3A mutations define a cytotoxic subset in PTCL-TBX21 with prognostic significance and thus may further refine pathological heterogeneity in PTCL-NOS and suggest alternative treatment strategies for this subset.

We tested the hypothesis that local inhibition of NOS would 1) alter the kinetics of vasodilation during dynamic forearm (FA) exercise and 2) prolong the time to reach steady state (SS). Subjects (n=22) performed rhythmic FA exercise (20% of maximum) during saline infusion (control) and NOS inhibition (L‐NMMA). Additionally, a subset of subjects performed a third trial of FA exercise during combined inhibition of either 1) NOS and adenosine (aminophylline; n=9), or 2) NOS and prostaglandins (Ketorolac; n=8). Forearm vascular conductance (FVC; ml/min/100mmHg) was calculated from blood flow (ml/min) and blood pressure (mmHg). Vasodilator kinetics were assessed by calculating the slope of the FVC response for every duty cycle between baseline and SS exercise as well as the number of duty cycles (1s contraction/2s relaxation) to reach SS. NOS inhibition reduced the kinetics of vasodilation during dynamic FA exercise (8.5 ± 0.6 vs. 11.1 ± 0.8 FVC/duty cycle; P < 0.001 vs. control) and increased the time to reach SS (25.1 ± 1 vs. 32 ± 1 duty cycles; P < 0.001 vs. control). The kinetics of vasodilation were reduced further with combined inhibition of NOS and adenosine (6.2 ± 0.8 vs. 7.5 ± 0.6 FVC/duty cycle; P < 0.05 vs. L‐NMMA alone), but not with combined inhibition of NOS and prostaglandins. Our data indicate that NOS inhibition blunts the kinetics of vasodilation during dynamic FA exercise and prolongs the time to reach SS.

Abstract Introduction: Recent results from two independent patient series have shown that chromosomal rearrangements of DUSP22 (DUSP22r+) and TP63 (TP63r+) can predict outcome in ALK-negative anaplastic large cell lymphoma (ALK-ALCL) and peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) with morphologic features resembling ALK-ALCL (Parilla-Castellar E, Blood 2014; Pedersen MB, Blood 2017). While DUSP22r+ is predictive for excellent survival similar to that of ALK+ALCL after CHOP/CHOP-like therapy, the rarely occurring TP63r+ is associated with an aggressive clinical behavior and poor outcome. The largest subgroup, i.e. patients with neither ALK nor DUSP22 nor TP63 rearrangements (triple negative), show a 5 year (yr) overall survival (OS) intermediate between that of ALK-/DUSP22r+ and ALK-/TP63r+ patients. The aim of the present study was to assess the impact of upfront high-dose therapy with autologous stem cell transplant (HDT/ASCT) on outcome in adult ALCL and PTCL-NOS patients according to their ALK, DUSP22 and TP63 status. The survival results from the two published series were pooled with those of a Nordic Lymphoma Group trial, the NLG-T-01 (d'Amore et al, JCO 2012), where patients were treated with 1st line CHOEP/CHOP followed, in chemosensitive cases, by upfront HDT/ASCT. Methods: Fluorescence in situ hybridization was performed on sections of previously constructed tissue microarrays using break-apart probes for the DUSP22-IRF4 and TP63 loci and a dual-fusion probe for TBL1XR1/TP63 fusion [inv(3)(q26q28)]. Evaluation of DUSP22 and TP63 rearrangements was performed in a blinded fashion without knowledge of PTCL subtype, clinical course, or outcome. Three independent patient cohorts were included: (i) one from Mayo Clinic consisting of 31 DUSP22r-, ALK-ALCL and PTCL-NOS (triple negative: 25; TP63r+: 6); (ii) one from Denmark consisting of 93 DUSP22r-, ALK-ALCL and PTCL-NOS (triple negative: 90; TP63r+: 3); and one from the NLG-T-01 trial consisting of 46 ALK-ALCL and PTCL-NOS (triple negative: 37; TP63r+: 1; DUSP22r+: 8), leading to a total study population of 170 patients. ALK+ ALCL was not included in the analysis, since no patients with this histology entered the NLG-T-01 trial. Association of genetic subtype with OS was assessed using Kaplan-Meier curves and Cox proportional models for hazard rate ratios (HR). Significant differences were defined as P &lt;0.05. Results: The eight DUSP22r+ patients (7 ALK-ALCL and 1 PTCL-NOS) from NLG-T-01 had a 5-yr OS of 83%, (95%CI 27-97), similar to that reported for DUSP22r+ in the Mayo and Danish cohorts (90% and 80%, respectively). No lymphoma-related events were observed in this subset. The only event was a septic death due to HDT-induced cytopenias in a patient who was in complete remission (CR). Among the 162 patients with DUSP22r-, ALK-ALCL and PTCL-NOS, those consolidated with HDT/ASCT (n=47) had a significantly better outcome (5-yr OS: 45%) than those treated with induction chemotherapy alone (5-yr OS: 30%) (n=115) (P=0.01). The patients in the HDT/ASCT group were younger (P&lt;0.001) and had more advanced stage disease (P=0.002). The improvement in OS persisted when the analysis was limited to transplant eligible patients and adjusted for age and stage (HR 0.52 95%CI 29-91, P=0.023) or International Prognostic Index (IPI) groups (HR 0.49, 95%CI 20-82, P=0.006). Of the 10 TP63r+ patients found within the pooled cohort, 6 were transplant eligible. Of these, 5 were not transplanted (intention-to-treat) while one was (Nordic trial). The latter patient achieved a still ongoing long-term CR, while all others died of lymphoma progression. Conclusion: In ALK-ALCL and PTCL-NOS patients from the NLG-T-01 trial, DUSP22r+ was associated with a very good outcome, similar to that seen in DUSP22r+ patients who had not undergone upfront autologous transplant. This observation supports the impression that upfront HDT/ASCT may not be of benefit in these patients. TP63r+ predicted poor outcome in non-transplanted patients. The impact of HDT/ASCT in the TP63r+ setting could not be adequately evaluated, since only one patient from the NLG-T-01 trial cohort was found to be TP63r+. Notably, this patient was the only survivor of the TP63r+ subset. For DUSP22r-, ALK-ALCL and PTCL-NOS patients taken as one group, those who received upfront HDT/ASCT had a superior survival compared to their age- and IPI-matched non-transplanted counterparts. Disclosures Ellin: ROCHE: Consultancy, Research Funding; CTI: Consultancy. Mannisto: Roche: Honoraria, Other: Travel expence; Takeda: Honoraria, Other: Travel expence; Amgen: Other: Travel expence; Novartis: Other: Travel expence; Celgene: Other: Travel expence; Gilead: Other: Travel expence; Pfizer: Honoraria; SOBI: Honoraria. Cerhan: Janssen: Other: Scientific Advisory Board (REMICADELYM4001); Janssen: Other: Multiple Myeloma Registry Steering . Toldbod: Takeda Pharma: Honoraria.

Abstract Background IL-10 polarized macrophages promote tumor growth and survival. The T-cell transcription factor GATA binding protein-3 (GATA3) epigenetically regulates IL10 expression. Therefore, we investigated the extent to which GATA-3 regulates IL-10 production and macrophage polarization in T-cell lymphomas. Methods Monocyte-derived macrophages were generated and polarized with T-cell lymphoma conditioned media ((TCL-CM). Immunophenotypic and functional characteristics associated with IL10-polarized macrophages were examined. GATA3-dependent cytokine production was evaluated in T-cell lymphoma lines by shRNA knockdown. Alternatively polarized macrophages and GATA3 expression were examined in a cohort of patients with peripheral T cell lymphoma, not otherwise specified (PTCL, NOS) (n=76). Comparisons among groups were evaluated using a Student's t test. Progression-free and overall survival (PFS & OS) was estimated using the Kaplan-Meier method and two-tailed log-rank test based on Cox proportional hazards model. Results To investigate the ability of T-cell lymphomas to alternatively polarize macrophages, monocytes from healthy donors were cocultured with TCL-CM. Macrophages were shown to have immunophenotypic characteristics of IL10-polarized macrophages (pSTAT3+/CD16hi/CD163hi/HLA-DR-/lo) by flow cytometry, which was abrogated by IL10, but not IL6, neutralization. TCL-CM polarized macrophages produced abundant IL-10 and were impaired in their ability to stimulate T-cell proliferation. Collectively, these results demonstrate that T-cell lymphomas promote alternative macrophage polarization in an IL-10 dependent manner, and both CD163 and pSTAT3 may aid in their identification in clinical T-cell lymphoma specimens. Therefore, CD163+/pSTAT3+ macrophages were visualized by immunofluorescence in PTCL, NOS specimens. IL-10 polarized macrophages were not identified in normal tissues, but were abundant constituents of the tumor microenvironment in 68% of the PTCL, NOS specimens examined (n=31). STAT3 phosphorylation in response to IL-10 is dependent upon the Janus kinases (JAK) JAK1 and TYK2, both of which are inhibited at nanomolar (and clinically achievable) concentrations by the “JAK2” inhibitor ruxolitinib. Therefore, we sought to determine whether ruxolitinib may prevent IL-10-induced macrophage polarization. As expected, ruxolitinib inhibited IL-10-induced STAT3 phosphorylation. More importantly, macrophages polarized in the presence of ruxolitinib lost the immunophenotype characteristics of IL-10 polarized macrophages, and were CD16-/loCD163-/loHLA-DRhi. Furthermore, IL-10 production was significantly impaired in macrophages polarized in the presence of ruxolitinib, while their ability to stimulate T-cell proliferation was significantly increased. In a separate initiative, we identified GATA3 as the key regulator of IL-10 and Th2 cytokine (IL-4, IL-13 and IL-5) expression in T-cell lymphoma lines by shRNA knockdown assays. Therefore, we examined GATA3 expression by immunohistochemistry in PTCL, NOS specimens. GATA-3 expression was observed in most of the T-cell lymphoma lines and in 47% of PTCL, NOS specimens (n=76). In the cohort of the 76 PTCL, NOS patients, the median PFS and OS observed in GATA3+ PTCL, NOS was 5 months (4-8 months, 95% CI) and 8 months (5-14 months, 95% CI), respectively. In contrast, the median PFS and OS in GATA3- PTCL, NOS was 1.3 years (0.6-1.6, 95% CI) and 1.6 years (0.7-6.9 95% CI), respectively. Conclusions Alternatively polarized macrophages are abundant in T-cell lymphomas and are generated in an IL-10- and STAT3-dependent manner. GATA-3 regulates IL-10 production in these lymphomas, and its expression identified a high-risk subset of PTCL, NOS with distinct clinicopathological features. As IL-10-dependent STAT3 phosphorylation and macrophage polarization were impaired by ruxolitinib, the JAK2 inhibitors may warrant closer scrutiny as immunomodulatory agents. Disclosures: Off Label Use: Ruxolitinib, a JAK2 inhibitor, may warrant closer scrutiny as an immunomodulatory agent in TCL therapeutics.