Xiaopei Wang

L-asparaginase in the treatment of refractory and relapsed extranodal NK/T-cell lymphoma, nasal type.

There is no standard salvage regimen for patients with refractory and relapsed extranodal NK/T-cell lymphoma (NKTCL), nasal type. This study was conduced to evaluate the efficacy of L-asparaginase-based regimen as a salvage regimen, on refractory and relapsed extranodal NKTCL, nasal type. Between March 1996 and March 2008, 45 patients with refractory and relapsed extranodal NKTCL, nasal type, were studied retrospectively. All patients were treated with L-asparaginase-based salvage regimen. Thirty-nine patients also received primary involved-field radiation after L-asparaginase-based chemotherapy. The complete response rate, partial response rate, and overall response rate for the whole group were 55.6%, 26.7%, and 82.2%, respectively. Both of 3-year and 5-year overall survival (OS) rates were 66.9%. The major adverse effects of L-asparaginase were myelosuppression, liver dysfunction, hyperglycemia, and allergic reaction. In general, the side effects could be tolerated. On univariate analysis, age, the stage of disease, and performance status were found to be prognostic factors influencing OS. On multivariate analysis, the stage of disease and age were independent prognostic factors for OS. L-Asparaginase-based regimen was obviously effective for the patients with refractory and relapsed extranodal NKTCL, nasal type.

Safety and efficacy of bevacizumab combined with R-CHOP regimen in seven Chinese patients with untreated diffuse large B-cell lymphoma.

BackgroundRituximab plus CHOP (R-CHOP) significantly improved the outcome of diffuse large B cell lymphoma (DLBCL), a common sub-type of non-Hodgkin lymphoma. But 40% – 50% of DLBCL patients cannot be cured by this regimen. Some clinical trials showed that bevacizumab might be useful in the treatment of DLBCL. This study evaluated the safety and efficacy of bevacizumab combined with the R-CHOP (A-R-CHOP) regimen in Chinese patients with previously untreated DLBCL.MethodsPatients with previously untreated DLBCL received A-R-CHOP regimen therapy. All patients with complete response (CR)/ unconfirmed complete response(CRu) after 8 cycles of A-R-CHOP received the bevacizumab maintenance therapy once every 3 weeks. The remained bulky disease was treated with radiotherapy.ResultsSeven Chinese patients were treated. All of them had bulky diseases. One patient had progressive disease after 4 cycles of A-R-CHOP therapy. The rest six patients completed 8 cycles of A-R-CHOP treatment. All of these six patients reached CR/CRu (5 CR, 1 CRu). Bevacizumab maintenance therapy was given to 4 CR patients. All 7 patients experienced Grade 3/4 hematologic adverse events; additionally, one had Grade 3 gastrointestinal toxicity and one had Grade 1 epistaxis. During bevacizumab maintenance therapy, one patient had Grade 1 gingival bleeding, another experienced Grade 1 proteinuria and then Grade 3 congestive heart failure 4 months after completion of maintenance therapy. At the end of July 2013, the patient who had progressive disease after 4 cycles of A-R-CHOP died of progressive disease, the other six remained CR response.ConclusionsThe A-R-CHOP regimen is effective for untreated DLBCL, but may cause bevacizumab-specific toxicities, which should be monitored.

Klotho suppresses tumor growth of T-cell lymphoma via inhibiting IGF-1R signaling

mutations, gene expression, and immune repertoires is challenging, as specific assays are required for each. We developed targeted NGS assays based on Anchored Multiplex PCR (AMP) that enable comprehensive mutation detection, gene expression analysis, and characterization of the expressed immune repertoire. Methods: AMP uses molecular barcodes (MBCs) ligated to cDNA ends and gene‐specific primers for amplification, enabling open‐ended sequencing of target regions to identify both known and unknown mutations, including novel gene fusions. MBC adapters ligated to RNA fragments prior to amplification enable relative gene expression analysis. Furthermore, open‐ended amplification enables immune chain mRNA interrogation from a single side, eliminating the need for opposing primers that bind within the highly variable V‐segment and thus preventing clone dropout due to somatic hypermutation. This also facilitates CDR3 sequence capture from highly fragmented RNA inputs. Results: We developed primer sets to detect known and unknown mutations and analyze relative gene expression levels in lymphoma. Here, we show the ability of this assay to detect characteristic IGH fusions as well as detect partner‐gene expression levels in diffuse large B‐cell lymphoma (DLBCL). Furthermore, this assay enabled NGS‐based expression profiling for identification of DLBCL subtypes in a small cohort of samples. We also developed the Archer Immunoverse assay for sequencing of the expressed B‐cell and T‐cell repertoire. We validated the quantitative reproducibility and sensitivity of the AMP‐ based IGH assay using mRNA isolated from peripheral blood leukocytes of healthy and B‐cell chronic lymphocytic leukemia (B‐ CLL) donors. Our data showed high reproducibility between replicates and quantitative clone tracking down to 0.01%, with the ability to determine IGHV mutational status. Furthermore, we identify a high‐ frequency B‐cell clones in FFPE DLBCL samples as well as hypermutation status. Conclusions: AMP‐based NGS enables detection of mutations, gene expression, and the expressed immune repertoire for comprehensive characterization and monitoring of malignant lymphoma.

MELK serves as a potent independent biomarker to predict survival of diffuse large B-cell lymphoma patients treated with rituximab-CHOP regimen

macodynamics of R‐CHOP (www.pharmgkb.org) selected and analysed in relation to R‐CHOP efficacy. Univariate and multivariate logistic regression analyses were performed to evaluate associations between SNPs and clinical/pathological characteristics or survival parameters (PFS and OS). Results: Median age was 63 years. There were 37 men and 43 women. A total of 47.5% of pts were in stage I‐II and 52.5% of pts in stage III‐IV. 27.5% of pts had bulky disease; 43.8% of pts had involvement of extranodal site. 47.5% of pts had pathological LDH value. According to the revised IPI, 15% of pts were in the low‐risk group, 58.7% in the intermediate, and 26.3% in the high‐risk group. Overall, 468 courses of R‐CHOP had been administered (mean: 5.85 courses, range: 4‐6). A total of 88,7% of pts had CR to R‐CHOP whereas the remaining showed PR or SD (7.5%) or PD (3,8%).Multivariate analysis identified FCGR2A rs1801274 as a predictor of PFS (P = .045). Pts with HR or RR genotypes showed shorter PFS than pts with HH genotype (HR: 2.437, 95% CI, 1.020‐5.823). No statistically significant correlation was found between SNPs and OS. Conclusions: Our preliminary data obtained in a limited number of pts show an association between a SNP of the low affinity FCGR2A gene involved in the activity of rituximab and PFS. Further insights will derive from the completion of pts accrual to reach the planned number of cases at the end of our study. Acknowledgements: This work was supported by a grant from the Associazione Giacomo Onlus, Castiglioncello (LI), Italy to E.M. and Cassa di Risparmio di Firenze, Firenze, Italy to S.N.

THE BRUTON'S TYROSINE KINASE INHIBITOR IBRUTINIB EXERTS IMMUNOMODULATORY ACTIVITY TOWARDS TUMOR-INFILTRATING MACROPHAGES

Introduction:Metformin exerts anti‐tumor effects in part by activating AMP‐activated protein kinase (AMPK) with resultant reduction in pro‐ survival mTOR signaling. Here, we test the hypothesis that metformin induces cell death by altering mitochondrial metabolism in lymphoma cell lines. As Bcl‐2 proteins, cyclin‐dependent kinases (CDK) and Phosphoinositol‐3‐kinase (PI3K) also influence mitochondrial physiology and metabolism, we also explored the additive effects of metformin with novel agents including the Bcl‐2 inhibitor (ABT‐199), the CDK9 inhibitor (BAY‐1143572) and the p110δ‐selective PI3K inhibitor (CAL‐101). Methods: Daudi (Burkitt), SUDHL‐4 (GC DLBCL), Jeko‐1 (MCL) and KPUM‐UH1 (double hit DLBCL) cells were plated in 96‐well plates and treated with varying doses of metformin (0–5000 μM). Hoechst 33342 DNA‐binding dye or MTS assays were used to measure proliferation and cell viability, respectively. Perturbations in oxidative and glycolytic metabolism were monitored via Seahorse XF Energy Phenotype kits. Lastly, efficacy of combination therapy was evaluated in cells co‐treated with metformin and ABT‐199 or BAY‐ 1143572 or CAL‐101. Results: Daudi and SUDHL‐4 cells showed 80% and 50% reduction in viability, respectively, with metformin at levels greater than 1 mM at day 7, while KPUM‐UH1 and Jeko‐1 cells were unaffected. The Seahorse analyses showed reduction in oxidative phosphorylation, in cells treated with 1mM metformin with a corresponding increase in glycolysis. Co‐treating KPUM‐UH1 and SUDHL‐4 cells with 10 mM metformin resulted in 1.4‐fold and 8.8‐fold decreases, respectively, in IC50 values of ABT‐199 at day 3 (Table 1). Jeko‐1 and Daudi cells were resistant to ABT‐199. By contrast, 3‐fold and 10 fold reduction in IC50 values of BAY‐1143572 in Daudi and Jeko‐1, respectively, was seen in the presence of 10 mM Metformin, and SUDHL‐4 or KPUM‐UH1 cells were unaffected. No change in IC50 value for CAL‐ 101 was observed. Conclusions: Metformin induces significant changes in cellular metabolism leading to decreased proliferation and viability in lymphoma cell lines. The combination of metformin and the anti‐ Bcl‐2 agent ABT‐199 achieves an additive effect in DLBCL (including double hit) cell lines, whereas the combination with CDK9 inhibitor BAY‐1143572 provides more selective additive anti‐tumor effect in Burkitt and MCL cell lines. These data support the idea of targeting cancer metabolism with metformin in novel combinatorial therapeutic strategies.

A PHASE I STUDY OF CHIMERIC ANTIGEN RECEPTORMODIFIED T CELLS DIRECTED AGAINST CD19 IN PATIENTS WITH RELAPSED OR REFRACTORYCD19(+) B CELL LYMPHOMAS: INTERIM ANALYSIS

271 A PHASE I STUDY OF CHIMERIC ANTIGEN RECEPTORMODIFIED T CELLS DIRECTED AGAINST CD19 IN PATIENTS WITH RELAPSED OR REFRACTORYCD19(+) B CELL LYMPHOMAS: INTERIM ANALYSIS Z. Ying* | X. Xiang | Y. Song | N. Ding | Y. Lin | W. Zheng | X. Wang | N. Lin | M. Tu | Y. Xie | C. Zhang | W. Liu | L. Deng | Y. Liu | Y. Yue | X. Yu | H. Liu | P. Duan | F. Chen | X. Wu | X.F. Huang | L. Jones | X. Kang | S. Chen | J. Zhu Department of Lymphoma, Peking University Cancer Hospital & Institute, Beijing, China; Department of medicine, Marino Biotechnology Co., Ltd., Beijing, China; Department of Molecular Microbiology and Immunology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, USA