Key Points• Activating mutations in PLCG1 are a frequent finding in tumoral CTCL samples. This raises the possibility of targeted therapies against PLCG1 signaling pathway, using calcineurin inhibitors.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 kB, 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. (Blood. 2014;123(13):2034-2043

Mutated JAK kinases and deregulated STAT activity are potential therapeutic targets in cutaneous T-cell lymphomaThe malignant mechanisms that control the development of cutaneous T-cell lymphoma (CTCL) are starting to be identified. Recent evidence suggests that disturbances in specific intracellular signaling pathways, such as RAS-MAPK, TCR-PLCG1-NFAT and JAK-STAT, can play an essential role in the pathogenesis of CTCL.1,2 Our group previously reported a network of somatic mutations affecting genes with potential to affect critical Tcell signaling pathways in CTCL patients. 1 As part of our findings we detected a number of mutations potentially affecting JAK/STAT signaling. These findings were recently confirmed by an independent group, suggesting that mutations in this pathway may contribute as disease mechanisms in CTCL.3 Deregulated JAK/STAT signaling is involved in many types of cancer. In fact, somatically acquired genetic alterations of JAK or STAT genes that induce aberrant activation of downstream signaling, via STAT phosphorylation, have been reported in some human hematologic malignancies including T-cell lymphomas. 4,5 We decided to explore JAK/STAT signaling as part of an intricate network of malignant signaling that controls the pathogenesis of CTCL, on the basis of the following evidence: (i) we had detected mutations in the pseudokinase domain of JAK1 and JAK3 in two of 11 patients and one cell line; (ii) we had also found several mutations that can directly (i.e., IL6S/T) or indirectly (i.e., TRAF6, RELB and CARD11) activate JAK/STAT signaling; and (iii) activated STAT3 had been detected in a large proportion of patients with advanced CTCL. 6,7 To explore the mutational status of JAK genes in a larger cohort of human CTCL patients' samples and cell lines, two independent state-of-the-art ultrasequencing approaches were used: a targeted gene-enrichment kit (HaloPlex) coupled to Ion-PGM (Life Technologies) sequencing, and a specific polymerase chain reactionbased amplification protocol targeting the pseudokinase domains of JAK1, JAK2 and JAK3 genes (hereafter, referred to as PsTKd-PCR), followed by specific indexing and sequencing with MiSeq (Illumina; see the Online Supplementary Methods for details). These are two highly sensitive methods that can enable the detection of mutations even present at low frequencies in neoplastic cells or in minority clones which may be found in CTCL samples. Thus, taken together, the data from our series (including those already described by Vaqué et al. © F e r r a t a S t o r t i F o u n d a t i o nthe pseudokinase domain of JAK proteins, a finding that is consistent with the results of other research groups that have found somatic mutations in the same domain of JAK1 and JAK3 kinases in prolymphocytic leukemia, other T-cell leukemias including CTCL and various human malignancies. 3,4,[8][9][10] Thus, it has been shown that JAK pseudokinase domains are auto-inhibitory and keep the kinase domain inactive until receptor dimerization stimulates transition to an a...

Methods: Observational, open-label multicenter study from 40 national referral centers of GCA patients treated with TCZ due to inefficacy or adverse events of previous therapy. Outcomes variables were improvement of clinical features, acute phase reactants, glucocorticoid-sparing effect, prolonged remission and relapses. A comparative study was performed: a) TCZ route (SC vs. IV); b) GCA duration (≤6 vs. >6 months); c) serious infections (with or without); d) ≤15 vs. >15 mg/day at TCZ onset. Results: 134 patients; mean age, 73.0±8.8 years. TCZ was started after a median [IQR] time from GCA diagnosis of 13.5 [5.0-33.5] months. Ninety-eight (73.1%) patients had received immunosuppressive agents. After 1 month of TCZ 93.9% experienced clinical improvement. Reduction of CRP from 1.7 [0.4-3.2] to 0.11 [0.05-0.5] mg/dL (p<0.0001), ESR from 33 [14.5-61] to 6 [2-12] mm/1 st hour (p<0.0001) and decrease in patients with anemia from 16.4% to 3.8% (p<0.0001) were observed. Regardless of administration route or disease duration, clinical improvement leading to remission at 6, 12, 18, 24 months was observed in 55.5%, 70.4%, 69.2% and 90% of patients. Most relevant adverse side-effect was serious infections (10.6/100 patients-year), associated with higher doses of prednisone during the first three months of therapy. Conclusion: In clinical practice, TCZ yields a rapid and maintained improvement of refractory GCA. Serious infections appear to be higher than in clinical trials.

Cystoid macular edema (CME) is a leading cause of blindness. In this study we assessed the efficacy and safety of Tocilizumab (TCZ) in refractory CME. DESIGN: Retrospective case series. METHODS: Patients with CME secondary to non-infectious uveitis who had inadequate response to corticosteroids and at least one conventional immunosuppressive drug, and in most cases to other biological agents were studied. CME was defined as central retinal thickness greater than 300 µm. The primary outcome measure was macular thickness. Intraocular inflammation, best corrected visual acuity (BCVA), and corticosteroid-sparing effect were also analyzed. RESULTS: A total of 25 patients (mean± SD age 33.6±18.9 years; 17 women) with CME were assessed. Underlying diseases associated with uveitis-related CME are juvenile idiopathic arthritis (n=9), Behçet's disease (n=7), birdshot retinochoroidopathy (n=4), idiopathic (n=4), and sarcoidosis (n=1). The ocular patterns were panuveitis (n=9), anterior uveitis (n=7), posterior uveitis (n=5) and intermediate uveitis (n=4). Most patients had CME in both eyes (n=24). TCZ was used in monotherapy (n=11) or combined with conventional immunosuppressive drugs. Regardless of the underlying disease, compared to baseline, a statistically significant improvement in macular thickness (415.7±177.2 vs 259.1±499.5 microns; p=0.00009) and BCVA (0.39±0.31 vs 0.54±0.33; p =0.0002) was obtained, allowing us to reduce the daily dose of prednisone (15.9±13.6 mg/day vs 3.1±2.3 p=0.002) after 12 months of therapy. Remission was achieved in 14 patients. Only minor side effects were observed after a mean follow up of 12.7±8.34 months. CONCLUSION: Macular thickness is reduced following administration of TCZ in refractory uveitis-related CME.

Inflammatory low back pain, pelvic girdle and diffuse lower limb pain are predictors of positive PET/CT scan for LVV in PMR.

No abstract

Our findings suggest that the presence of 6p25.3 rearrangement might be related to this particular biphasic pattern.

DUSP22-rearranged anaplastic lymphomas are characterized by specific morphological features and a lack of cytotoxic and JAK/STAT surrogate markers ALK-negative anaplastic large cell lymphoma (ALKnegative ALCL) is a heterogeneous disease with very disparate outcomes. Molecular studies have identified chromosomal rearrangements involving the DUSP22-IRF4 locus on 6p25.3 (DUSP22 rearrangements) as a favorable prognostic factor, associated with complete remission after first treatment thereby suggesting that this subgroup of patients may not gain additional benefit from autologous stem cell transplantation in first remission. [1][2][3] Recognition of these cases is critical, and we therefore aimed to study in greater detail the histological and immunophenotypic features of DUSP22-rearranged ALK-negative ALCLs.After approval by the Institutional Review Board of the Hospital Universitario Marqués de Valdecilla and the Fundación Jiménez Díaz, Spain, we collected 91 cases with a diagnosis of systemic or primary cutaneous ALCL made at the participating institutions. Clinical data were retrieved and cases were reviewed by 3 independent pathologists (AO, SMRP, and MAP) using hematoxylin & eosin stains. Immunohistochemistry was performed using a panel of antibodies against ALK, CD3, CD4, CD8, granzyme B, MUM1, perforin, P-STAT3 (D3A7, 1/400 Cell Signaling), TIA1, P-STAT5, TCR-βF1, P63, STAT3 (Online Supplementary Appendix). Of 91 evaluated cases, 18 were primary cutaneous ALCLs (pcALCLs) and 73 cases were systemic ALCLs (19 were ALK-positive ALCLs). ALK-positive cases were not further considered for the study. Only 31 cases were eligible for further study due to tissue scarcity, including 22 ALK-negative ALCL and 9 pcALCLs. Fluorescence in situ hybridization (FISH) analyses were performed on these cases using an IRF4-DUSP22 (6p25.3) break-apart probe (KBI-10613; Kreatech, Leica, Spain) following standard procedures. 4,5 Cytotoxic markers, pSTAT3, p63 and MUM1 expression were evaluated as described in the Online Supplementary Appendix. Associations of genetic and immunohistochemical subgroups with overall survival (OS) and progression-free survival were assessed using Kaplan-Meier curves. Differences between genetic subgroups in patients' characteristics, tumor phenotype and other clinical factors were assessed using the χ 2 test and Wilcoxon rank-sum test, as appropriate.