T-cell non-Hodgkin's Lymphoma Patients Research Articles

Phase 1 Clinical Trial of Memory-Enriched Academic HSP-CAR30 for the Treatment of Relapsed/Refractory Hodgkin Lymphoma and CD30+ T-Cell Lymphoma: Clinical and Biological Studies

Background: Patients with classical Hodgkin lymphoma (cHL) relapsed/refractory to immunotherapies (e.g., brentuximab, anti-PD-1 antibodies) have poor outcome. We developed an academic CART30 (HSP-CAR30) targeting a proximal epitope within the CD30 molecule to overcome soluble CD30. We generated products enriched in memory T-cells to ensure persistence, and enhancement of antitumor efficacy (Alvarez-Fernandez et al, 2021). Here, we report the results of our Phase 1 study evaluating HSP-CAR30 for the treatment of R/R cHL and CD30+T-cell non-Hodgkin lymphoma (T-NHL) (NCT04653649). Aim: Primary endpoints were to assess safety of HSP-CAR30 and to establish maximum tolerated dose (MTD) recommended for the following phase 2. Secondary objectives included response rates and cellular kinetics. Methods: We conducted a phase 1 dose-escalation study in 11 patients with R/R HL or CD30+ T-NHL. cHL patients were R/R to treatments including chemotherapy, brentuximab, and anti-PD-1 antibodies, while T-NHL patients were R/R to at least 2 chemotherapy treatments. T-cells isolated from fresh leukapheresis of enrolled patients were transduced with a lentivirus encoding a second-generation 4-1BB-costimulated CAR, containing a scFv directed against an epitope within the proximal non-cleavable part of CD30 protein. Three cell-dose levels in three cohorts of patients (3+3 design) were evaluated: DL1 (3x106/kg), DL2 (5x106/kg) and DL3 (10x106/kg) CAR30+ T-cells. Results: From February 2021 to December 2021, 11 patients (9 cHL and 2 CD30+ T-NHL (1 ALK- ALCL and 1 PTCL NOS) were enrolled and underwent leukapheresis. Of these, 10 patients received HSP-CAR30: 3 patients at DL1, 3 at DL2 and 4 at DL3. Demographic characteristics and baseline disease features are summarized in Table 1. Median age was 50 years (range 21-65). Median number of prior lines of treatment was 5 (range 3-7). One patient was not infused due to lack of T-cell expansion. All patients received lymphodepletion before infusion: fludarabine/bendamustine for cHL patients (n=8) and fludarabine/cyclophosphamide for T-NHL (n=2). Mean HSP-CAR30 expression of T-cell products was 94.79±1.07% (±SEM). Infusion products had a high proportion of CAR30+ memory T-cells: 26.09±6.06% memory stem (TSCM) and 67.52±6.07% central memory (TCM) in CD4+, while TSCM represented 42.2±5.4%, and TCM 50.5±5.7% of CD8+ CAR30 T-cells. HSP-CAR30 was well tolerated; there were no dose limiting toxicities (DLTs). Relevant adverse events are shown in the Table 2. Cytokine release syndrome (CRS) was observed in 6 (60%) patients (all grade 1). No patient developed neurotoxicity. Self-limited skin rash was seen in 4 (40%) patients. One patient with history of cytomegalovirus (CMV) infections had CMV pneumonia. Another patient developed pulmonary tuberculosis. Long-lasting cytopenias (>3 months) occurred in 2 patients, with a complete recovery at 5 months in one of them. The other was diagnosed with myeloid neoplasm post-cytotoxic therapy (MN-pCT) with complex karyotype. Best objective response was 100%, including 5 (50%) patients with complete response (CR), all with HL (DL1=1; DL2=3; DL3= 1). At data cutoff (July 25th, 2022), mean follow-up was 315 days (range 120-514) and mean PFS was 235 days (77- 444). All CRs were maintained during follow-up. Four patients died, 3 of them due to progressive disease (2 T-NHL and 1 cHL) and one with refractory MN-pCT while in CR of his cHL. Mean time to HSP-CAR30 cell peak level across all doses was 21.5 days (range, 4-63). At the time of in vivo HSP-CAR30 expansion, memory T-cells (i.e., TSCM and TCM) were the dominant cell subset within CD4+ T-cells (TSCM 20.7±5% and TCM 56.33±6.2% of CAR30+ T-cells) and represented a high proportion of CD8+ T-cells (TSCM 18.36±4.9% and TCM 30.27±8.1% of CAR30+ T-cells). HSP-CAR30 was detected in blood samples by qPCR in all patients at 4 months and in all evaluable patients (n=7) at 9 months. Conclusions: This is the first European academic CART clinical trial evaluating a T-cell memory-enriched CART 30. Our Phase 1 study provides evidence for feasibility and safety of HSP-CAR30. Additionally, HSP-CAR30 has shown promising efficacy in heavily treated cHL patient population, and this is being explored in the phase 2. Less-differentiated memory T-cells (TSCM and TCM) predominate at the time of in vivo HSP-CAR30 expansion. Updated clinical data and cellular kinetics studies will be presented at the meeting. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

S257: A PHASE 1, FIRST-IN-HUMAN, DOSE-ESCALATION CLINICAL TRIAL OF MEMORY-ENRICHED CD30-CAR T-CELL THERAPY FOR THE TREATMENT OF RELAPSED OR REFRACTORY HODGKIN LYMPHOMA AND CD30+ T-CELL LYMPHOMA

Background: Up to 30% of Hodgkin lymphoma (HL) patients are refractory or relapse (R/R) after first treatment and their prognosis is poor. We have developed a refined CD30-CAR-T (HSP-CAR30) targeting a proximal epitope within the CD30 molecule to overcome soluble CD30 and generated products enriched in memory T-cells to ensure efficient engraftment, persistence and enhancement of antitumor efficacy (Alvarez-Fernandez et al, 2021). Here, we report the results of our Phase 1 study evaluating HSP-CAR30 for the treatment of R/R HL and CD30+ T-cell non-Hodgkin lymphoma (T-NHL) (NCT04653649). Aims: Primary endpoints were to assess safety of HSP-CAR30 and to establish maximum tolerated dose (MTD) recommended for the following Phase 2. Secondary objectives include best response rates after infusion. Methods: We conducted a phase 1 dose-escalation study in 11 patients with R/R HL or CD30+ T-NHL. HL patients were R/R to treatments including chemotherapy, brentuximab and anti-PD-1 antibodies, while T-NHL patients were R/R to at least 2 chemotherapy treatments. T-cells were transduced with a lentivirus encoding a second-generation 4-1BB costimulated CAR, containing a scFv directed against an epitope from the proximal non-cleavable part of CD30 protein. Three cell-dose levels were evaluated: DL1 (3x106/kg), DL2 (5x106/kg) and DL3 (10x106/kg) CAR30+ T-cells. Results: From February 2021 to December 2021, 11 patients (9 HL and 2 CD30+ T-NHL) were enrolled and underwent leukapheresis. Of these, 10 patients received HSP-CAR30: 3 patients at DL1, 3 at DL2 and 4 at DL3. Demographic characteristics and baseline disease features are summarized in the Table. Median age was 49.9 years (range 21–65). Median number of prior lines of treatment was 4.6 (range 3–7). One patient did not received treatment due to lack of T-cell expansion. All patients received LD before infusion, fludarabine/bendamustine in HL patients (n=8) and fludarabine/cyclophosphamide in T-NHL (n=2). Mean HSP-CAR30 expression was 94.79±3,38% (±SD). Memory T-cell subset comprised 93.07±4,8% (±SD) in CD4+ and 91.64±4,9% (±SD) in CD8+. Mean time to HSP-CAR30 cell peak level across all doses was 29 days (range 6–98). CAR+ T-cells were detectable by flow cytometry up to 11 months after infusion. HSP-CAR30 infusion was well tolerated; there were no dose limiting toxicities (DLTs). Relevant adverse events are shown in the Table. Grade 1 cytokine release syndrome (CRS) was observed in 6 (60%) patients. No patient developed neurotoxicity. Self-limited skin rash was seen in 4 (40%) patients. One patient with history of cytomegalovirus (CMV) infections had CMV pneumonia. Another patient developed pulmonary tuberculosis. At data cutoff (February 21st, 2021), the median follow-up was 204 days (60–351). Best objective response was 100%, including 5 (50%) patients with complete response (CR), all with HL (DL1=1; DL2=3; DL3= 1). Three patients have died of progressive disease (2 T-NHL and 1 HL). There were no non-relapse mortality events. Median PFS and median overall survival (OS) was not reached. Six-month PFS for HL patients was 75%. Image:Summary/Conclusion: This is the first European academic CART clinical trial evaluating a T-cell memory-enriched CART 30. Our Phase 1 study provides evidence for feasibility and safety of HSP-CAR30. Additionally, HSP-CAR30 has shown promising efficacy in heavily treated HL patients that is being explored in a phase 2 trial already started.

Autologous Stem Cell Transplantation (ASCT) for T-Cell Non Hodgkin Lymphoma (T-NHL) Patients Who Achieve Complete Remission with First-Line Treatment: A Systematic Review and Meta-Analysis

Introduction: There is currently no consensus on the optimal frontline therapy for patients with T cell Non-Hodgkin lymphomas (T-NHL). Consolidative autologous stem cell transplant (ASCT) is frequently offered to the patients with chemosensitive disease based on retrospective and prospective studies showing improved progression-free survival (PFS) when compared with historical controls getting chemotherapy alone. However, it remains unclear whether there is a good risk subset of patients who achieve first complete remission (CR1) following induction chemotherapy and who might not benefit from upfront ASCT. To date, no randomized control trials (RCTs) exist and available data is conflicting. We perform a systematic review/meta-analysis of the published literature to address this question. Methods A comprehensive, systematic search (from database inception - 9/2019) of MEDLINE/PubMed, EMBASE and Cochrane databases was performed. PRISMA and Meta-Analysis Of Observational Studies in Epidemiology (MOOSE) guidelines were followed. Studies were selected from a total of 2656, screened based on predefined inclusion/exclusion criteria, and were critically appraised for outcomes of interest [progression free survival (PFS) and overall survival (OS)]. Quality of studies was assessed using Newcastle-Ottawa Scale. Hazard ratios (HRs) and corresponding 95% Cis were calculated, and the meta-analysis was performed using the random-effects model. Test for heterogeneity was performed using I2 statistic. Results Of 2656 unique records, 13 studies (prospective = 3; retrospective = 10) were selected. In 8 studies, upfront ASCT was compared to NO ASCT in patients in first complete remission (CR1), while in 5, comparison was with patients achieving either PR1 (first partial remission) or CR1. 11 (of 13) studies reported PFS. Median follow-up in these studies ranged from 22 months to 7.8 years. Results from the meta-analysis showed that T-NHL patients who underwent ASCT had an improved 5-year PFS compared to those with NO ASCT (HR 1.62, 95% confidence interval (CI) 1.22 to 2.15, I² = 38%) (Figure 1). However, no benefit was observed in 5-year OS when ASCT was compared to NO ASCT (HR 1.32, 95% CI 0.82 to 2.12, I2 = 83%) (Figure 2). A sensitivity analysis including only studies with patients transplanted in CR1 showed similar findings, with a 5-year PFS (HR 1.54, 95% confidence interval (CI) 1.01 to 2.34, I² = 38%) and 5-yr OS (HR 1.11, 95% CI 0.41 to 3.02, I2 = 90%) when compared to No ASCT. Conclusions In the absence of RCTs, the results of this systematic review/meta-analysis represents the best evidence supporting long term PFS benefits of upfront ASCT consolidation in patients with T-NHL in CR1 or CR1/PR1 after frontline chemotherapy. Disclosures No relevant conflicts of interest to declare.

Predicting worse survival for newly diagnosed T cell lymphoma based on the decreased baseline CD16\u2212/CD16\u2009+\u2009monocyte ratio

T cell non-Hodgkin lymphoma (T-NHL) is highly invasive and heterogeneous without accurate prognosis prediction. We proposed peripheral CD16−/CD16 + monocytes the additional indicators for T-NHL prognosis. We prospectively recruited 31 T-NHL patients without previous treatment. The CD16−/CD16 + monocyte ratio before chemotherapy was calculated and regular follow up was performed to calculate prognostic prediction value. Tumor associated macrophages (TAM) in tumor tissue were counted and transcriptome sequencing of CD16− and CD16 + monocytes was applied to explore potential mechanisms. We found that T-NHL patients had higher ratio of total monocytes especially the CD16 + monocytes along with a decreased ratio of CD16−/CD16 + monocytes, compared to the health control. The 1-year overall survival rate was 0.492 and 0.755 for CD16− monocyte/CD16 + monocyte ratio of <11 and ≥11(p < 0.05), respectively. The peripheral CD16−/CD16 + monocyte ratio was significantly relevant with the pathological CD68/CD206 macrophage ratio. The differently expressed genes in CD16− and CD16 + monocytes from T-NHL patients were mainly involved in signaling molecules related to tumor microenvironment. Pro-tumor genes were identified in monocyte subsets especially in CD16 + monocytes. In conclusion, the ratio of peripheral CD16−/CD16 + monocyte helps to stratify the prognosis of T-NHL. The relatively increased CD16 + monocytes may contribute to the pro-tumor microenvironment of T-NHL.

Refined Disease Risk Index for Hematological Malignancies, Including Rare Disorders, After Allogeneic Stem Cell Transplantation

ObjectiveThe refined disease risk index (R-DRI) is a well-designed prognostic parameter that is based on only the disease type and status and is used for stratifying patients undergoing allogeneic hematopoietic stem cell transplantation (allo HSCT) into 4 risk groups. However, the application of the R-DRI for rare diseases has remained unclear. MethodsWe evaluated 135 patients who underwent allo HSCT for hematological malignancies including rare diseases, such as acute leukemia of ambiguous lineage, acute T-cell leukemia/lymphoma, extranodal natural killer T-cell lymphoma, and lymphoblastic lymphoma, at our institute. ResultsAccording to the R-DRI, overall survival (OS) and progression-free survival at 2 years for patients with the low, intermediate, high, and very high groups were 66.7% and 66.7%, 60.8% and 56.0%, 27.1% and 23.7%, and 5.9% and 5.1%, respectively (P < .0001 and P < .0001, respectively). OS showed no significant difference between B-cell non-Hodgkin lymphoma (B-NHL) and T-cell non-Hodgkin lymphoma (T-NHL) (P = .71). Moreover, OS at 1 year was 80%, 14.3%, 60%, and 0% for the intermediate risk group, the very high-risk group of B-NHL, the intermediate risk group, and the high-risk group of T-NHL, respectively (P = .035). ConclusionWe showed the applicability of the R-DRI for hematological malignancies, including rare disorders. However, we suggest that T-NHL patients may be better to be assigned between the nodal group and the extranodal group in the R-DRI.

Allogeneic Stem Cell Transplantation from Unmanipulated Haploidentical Donors Versus Unrelated Cord Blood in Patients with T-Cell Non-Hodgkin Lymphoma : A Retrospective Study from the Societe Francophone De Greffe De Moelle Et De Therapie Cellulaire (SFGMC-TC)

Introduction. Allogeneic stem cell transplantation (SCT) can be curative for those with high-risk or recurrent hematologic cancers including T-cell non-Hodgkin's lymphoma (TCL). For patients lacking an HLA-matched related donor, donor sources such as haplo-identical (Haplo) relatives or cord blood (UCB) represent reasonable alternatives. There is currently little data on the outcome of TCL patients undergoing alternative donor transplantation. The Société Francophone de Greffe de Moelle et de Transplantation Cellulaire (SFGM-TC) conducted this study to evaluate the outcome of recipients of alternate donor allo-SCT for TCL. Methods. 95 patients with non epidermotropic TCL, transplanted between 2003 and 2017, reported to ProMISE (Project Manager Internet Server), an internet-based system shared by 36 Francophone transplantation centers, were reviewed. Comparisons of outcome after transplants from different donors (Haplo, n=41 vs UCB, n=54) were performed with regard to overall survival (OS), non-relapse mortality (NRM), relapse and acute/chronic graft-versus-host disease (aGVHD/cGVHD) incidence. Results. Patients' characteristics are detailed in Table 1. Median age at HSCT was 42.5 (range, 3.6 - 68) years, with an older cohort for the Haplo group (50.7 vs 33.6 for UCB, p&lt; .001). Except for the median year of transplantation (median year 2017 for Haplo vs 2010 for UCB,) and the myeloablative regimen (22% for Haplo vs 48% for UCB, p .009), both Haplo and UCB groups were comparable concerning interval from diagnosis to HSCT, donor/recipient CMV status, sex and ABO mismatch. Patients received a median of 2 (range, 1-5 for Haplo; 1-6 for UCB) therapeutic lines. Twenty-eight (10 in Haplo vs 18 in UCB, p NS) underwent auto- or allo-SCT prior to Haplo or UCB-HSCT. Median time to engraftment was shorter (19 vs 25 days, p .025) in Haplo recipients. According to donor type, no differences were found in univariate analysis for the CI of grade II-IV aGVHD (35% vs 35%, p .88) and cGVHD (30% vs 28%, p .49). Median follow-up was 24 months and 47 months for Haplo and UCB recipients, respectively (p .004). At 12 months, OS was 71% and 50% (p .033), progression-free survival (PFS) 63% and 44% (p .045) and NMR 7% and 26% (p .014) in HSCT from Haplo and UCB, respectively. Relapse incidence was similar following HSCT from Haplo and UCB (32% vs 31%). Conclusion. Based on these first results, our study suggests that Haplo-HSCT is a strategy achieving prolonged survival as compared to UCB donors, due to less toxicity and similar incidence of relapse. An analysis of influencing factors will be further detailed. Figure Disclosures Blaise: Jazz Pharmaceuticals: Honoraria; Pierre Fabre medicaments: Honoraria; Molmed: Consultancy, Honoraria; Sanofi: Honoraria. Socie:Alexion: Consultancy. Mohty:Jazz Pharmaceuticals: Honoraria, Research Funding. Robin:Novartis Neovii: Research Funding.

Beneficial effect of consolidative radiotherapy for patients with lymphoma and skeletal involvement.

The objectives of this study were to analyze the clinical features of patients with bone involved lymphoma and identify the prognostic factors and to explore the optimized treatment strategy for bone involved lymphoma.A total of 1948 patients with lymphoma in our cancer center from September 2006 to October 2017 were retrospectively evaluated. Among these, 109 patients with skeletal involvement in lymphoma were enrolled. According to the pathologic subtypes, the patients were divided into 3 subgroups: classic Hodgkin lymphoma (cHL), B-cell non-Hodgkin lymphoma (B-NHL), and T-cell non-Hodgkin lymphoma (T-NHL). The clinical characteristics and overall survival (OS) of 3 groups of patients were reviewed, and the prognostic factors were analyzed.There were 9 (3 unifocal, 6 multifocal) patients with primary bone lymphoma. The 5-year OS of cHL, B-NHL, and T-NHL patients was 88.24%, 54.09%, and 61.58%, respectively. Advanced stage, elevated lactate dehydrogenase (LDH), age above 60, high International Prognostic Index score, and treatment without radiotherapy for the bone involved were significant poor prognostic factors for OS of all patients in univariate analysis. There was a trend toward better OS not only in limited-stage but also in advanced-stage patients with radiotherapy for the bone involved compared with the patients without radiotherapy. Elevated LDH level and age above 60 were the independent unfavorable prognostic factor in multivariate analysis.Elevated LDH level and age above 60 predict the poor prognosis of patients with bone involvement. The potential for long-term survival suggests that additional consolidative radiotherapy for the site of skeleton involvement may have a better chance of long-term success.

Autologous transplantation as consolidation for high risk aggressive T-cell non-Hodgkin lymphoma: a SWOG 9704 intergroup trial subgroup analysis

Phase II data suggest a benefit to autotransplantation for aggressive T-cell non-Hodgkin lymphoma (T-NHL) in first remission; randomized trials have yet to validate this. We performed a retrospective analysis of aggressive T-NHL patients in the intergroup randomized consolidative autotransplant trial (SWOG 9704). Of the 370 enrolled, 40 had T-NHL: 12 were not randomized due to ineligibility (n = 1), choice (n = 2), or progression (n = 9), leaving 13 randomized to control and 15 to autologous stem cell transplantation (ASCT). Two ASCT patients refused transplant and one failed mobilization. The 5-year landmark PFS/OS estimates for ASCT vs. control groups were 40% vs. 38% (p = .56), and 40% vs. 45% (p = .98), respectively. No difference was seen based on IPI, or histologic subtype. Only 1/7 receiving BCNU-based therapy survived vs. 4/5 receiving TBI. Aggressive T-NHL autotransplanted in first remission did not appear to benefit from consolidative ASCT. This and the 30% who dropped out pre-randomization mostly to progression, suggests that improved induction regimens be developed.

SPARC is down-regulated by DNA methylation and functions as a tumor suppressor in T-cell lymphoma

The aim of this study was to assess the functional role of SPARC in T-cell non-Hodgkin's lymphoma (T-NHL), as well as the underlying molecular mechanisms. Here, we first identified SPARC expression in T-NHL tissues and cell lines through western blot and real-time PCR (RT-PCR). Overall survival of T-NHL patients with different levels of SPARC was assessed by Kaplan-Meier survival curves. Then cell proliferation, apoptosis, migration and invasion of T-NHL cells with either knockdown or overexpression of SPARC were determined by MTT, flow cytometry, transwell migration and invasion assay, respectively. Finally, the molecular mechanism by which SPARC modulated T-NHL cell progression was assessed. We confirmed that SPARC was significantly down-regulated in T-NHL tissues and cell lines. T-NHL patients with high levels of SPARC demonstrated a favorable clinical outcome. SPARC significantly suppressed cell proliferation, migration and invasion, and EMT process, but facilitated cell apoptosis in T-NHL cells. Further, we found that loss of SPARC expression in T-NHL tissues and cell lines, both in mRNA and protein levels, was associated with the aberrant DNA methylation in SPRAC gene, and the disrupted SPARC expression could be rescued after treatment with the demethylating agent 5-Aza-2′-deoxycitydine (5-Aza-Cdr). Additionally, 5-Aza-Cdr reversed SPARC hypermethylation to restore its biological role as a tumor suppressor in T-NHL cells, including inhibiting cell proliferation, invasion and migration, while promoting cell apoptosis. Our data provided evidence that DNA methylation in SPARC gene may play a role in the progression of T-NHL.

Elevated monoclonal and polyclonal serum immunoglobulin free light chain as prognostic factors in B- and T-cell non-Hodgkin lymphoma.

The serum immunoglobulin free light chain (FLC) assay quantitates free kappa (κ) and lambda (λ) light chains. FLC elevations in patients with diffuse large B-cell lymphoma (DLBCL), Hodgkin lymphoma (HL), and chronic lymphocytic leukemia (CLL) are associated with an inferior survival. These increases in FLC can be monoclonal (as in myeloma) or polyclonal. The goal was to estimate the frequency of these elevations within distinct types of B-cell and T-cell non-Hodgkin lymphoma (NHL) and whether the FLC measurements are associated with event-free survival (EFS). We studied serum for FLC abnormalities using normal laboratory reference ranges to define an elevated κ or λ FLC. Elevations were further classified as polyclonal or monoclonal. Four hundred ninety-two patients were studied: 453 B-cell and 34 T-cell NHL patients. Twenty-nine % (142/453) of patients had an elevated FLC of which 10% were monoclonal elevations. Within B-cell NHL, FLC abnormalities were most common in lymphoplasmacytic (79%), mantle cell (68%), and lymphomas of mucosa associated lymphoid tissue (31%); they were least common in follicular (15%). The hazard ratio (HR) for EFS in all patients was 1.41 (95% CI; 1.11-1.81); in all B-cell NHL the HR was 1.44 (95% CI 1.11-1.96); in all T-cell NHL the HR was 1.17 (95% CI 0.55-2.49). FLC abnormalities predicted an inferior OS (HR = 2.75, 95% CI: 1.93-3.90, P < 0.0001). The serum FLC assay is useful for prognosis in both B-cell and T-cell types of NHL. In B-cell NHL further discrimination between a monoclonal and polyclonal elevation may be helpful and should be analyzed in prospective clinical trials.

Thalidomide plus chemotherapy exhibit enhanced efficacy in the clinical treatment of T-cell non-Hodgkin's lymphoma: A prospective study of 46 cases.

The treatment of T-cell non-Hodgkin's lymphoma (T-NHL) remains challenging. There is currently no standard regimen for the treatment of T-NHL in the first- or second-line setting. Thalidomide was previously shown to exert antitumor effects through inhibiting angiogenesis, promoting apoptosis and immunomodulatory activity. However, all the previous studies on the treatment of lymphoma with thalidomide included patient samples of limited size. In the present study, 46 cases of eligible T-NHL patients were randomized into i) the control group (conventional combined chemotherapy, n=22) and ii) the thalidomide group (thalidomide plus combined chemotherapy, n=24). The median dose of thalidomide was 200 mg (range, 150-400 mg) every night, without reported severe side effects. The clinical response to treatment was as follows: Complete response (CR) in 12 cases, partial response (PR) in 7, stable disease (SD) in 1 and progressive disease (PD) in 4 cases in the thalidomide group; and CR in 8 cases, PR in 6, SD in 3 and PD in 5 cases in the control group. The CR rate was 50.0 and 36.4% in the thalidomide and the control groups, respectively (P<0.05). The median progression-free and overall survival were 12 and undefined months, respectively, in the thalidomide group and 6 and 17 months, respectively, in the control group. The toxicity profile was considered acceptable in both groups. Our results indicated that thalidomide plus combined chemotherapy may exhibit enhanced efficacy in the clinical treatment of T-NHL. In addition, this type of treatment may reduce the frequency of adverse gastrointestinal reactions and help alleviate fear of chemotherapy. Therefore, thalidomide plus combined chemotherapy may be a viable option for the clinical treatment of T-NHL.

Expression of AEG-1 in human T-cell lymphoma enhances the risk of progression

The aim of this study was to examine the expression and role of astrocyte elevated gene-1 (AEG-1) in biological processes of T-cell non-Hodgkin's lymphoma (T-NHL). AEG-1 expression in T-NHL patients was characterized with immunohistochemistry. The expression of AEG-1, survivin, Bcl-2 and Bax in Jurkat and Hut-78 cells was detected by real-time PCR and western blotting. Cell proliferation, cell cycle and apoptosis were measured by MTT and flow cytometry. MMP-2/-9 activity was detected by gelatin zymography. Of the studied tumors, 104 (80.62%) exhibited cytoplasmic AEG-1 immunostaining. AEG-1-siRNA in Jurkat and Hut-78 cells suppressed cell proliferation and induced cell apoptosis, inhibited survivin and Bcl-2/Bax protein expression as well as MMP-2/-9 activity. Downregulation of AEG-1 using siRNA could provide a potential approach for gene therapy against T-NHL, and the antitumor effects may be associated with inhibition of survivin and Bcl-2/Bax protein expression and MMP-2/-9 activity.

LEF1 Gene Mutations In Childhood T-Cell Acute Lymphoblastic Leukemia and T Cell Non-Hodgkin's Lymphoma

AbstractAbstract 276LEF1 is a member of the LEF/TCF family of DNA-binding transcription factors, which interact with beta-catenin in the WNT signaling pathway. N-terminal LEF1 mutations that impair beta-catenin binding are commonly found inhuman sebaceous skin tumors, and expression of an N-terminal–deleted Lef1 mutant that lacks the beta-catenin binding domain leads to sebaceous skin tumors. The intracellular domain of NOTCH1 has also been shown to function as a co-activator of LEF1, leading to the up-regulation of target genes distinct from those activated by beta-catenin binding. Recently, inactivating mutations of LEF gene have been reported in T-cell acute lymphoblastic leukemia (T-ALL) cases. In this study, we analyzed the frequencies and clinical significance of LEF1 mutations in pediatric T-ALL and T-cell non-Hodgkin's lymphoma (T-NHL). Mutation analysis was performed on 138 of the primary T-ALL and T-NHL patient samples. The clinical data were available 55 newly diagnosed T-ALL and 14 T-NHL patients. These children, aged under 15 years were enrolled into the Japan Association of Childhood Leukemia Study (JACLS) protocol ALL-97 between 1997–2001 and JACLS trial NHL-T98 between 1998–2002. At the time of diagnosis, bone marrow and/or peripheral blood cells were obtained from T-ALL patients and lymph nodes and/or pleural effusions were obtained from T-NHL patients. A total of 69 patients were included in the present study; 49 were male and 20 female; 55 were children diagnosed with T-ALL (median age of 9.5 years; range: 2.0 – 15.0 years) and 14 with T-NHL (median age of 11.0 years; range: 3.7 – 15.0 years). We performed high-resolution array comparative genomic hybridization (array CGH) and sequencing was performed on the entire coding region of LEF1. High molecular weight genomic DNA was used for microarray analysis using Affymetrix GeneChip 50K XbaI, HindIII or 250K NspI, according to the manufacturer's instructions. Genome-wide detection of allelic imbalances was performed using CNAG/AsCNAR software. We identified mono or biallelic LEF1 deletions in 7.5% (5 of 67) of these primary samples. An additional 11.6% (16 of 138) of the cases harbored sequence alterations of LEF1 gene. Twelve of the single nucleotide alterations were found in exons 1, 2 and 3, and four of the frame-shift mutations were found in exons 3, 7, and 8. Frame-shift mutations were novel LEF1 mutations, and mutations in exons 3 and 8 were positioned outside catalytic domain and DNA binding region. Missense mutations in exon 1 were located highly conservative lesion of beta-catenin binding domain of LEF1. Of the 8 LEF1 alterations detected in 16 cases also had NOTCH1 mutations. Analysis of the available clinical data showed that LEF1 gene alteration was not a significant predictor of event-free survival in children with T-ALL treated in JACLS ALL-97 and NHL-T98 protocol. However, LEF1 inactivation was associated with a younger age at the time of diagnosis, but not with sex, white blood cell count, central nervous system involvement, or the presence of an anterior mediastinal mass at the time of diagnosis. Furthermore, analysis of the T-ALL cell surface immunophenotype obtained at the time of diagnosis showed that all LEF1-alterd cases were characterized by developmental arrest at a cortical stage of T-cell differentiation. The clinical significance of these gene alterations and detailed data is discussed. Disclosures:No relevant conflicts of interest to declare.

FBXW7 and NOTCH1 mutations in childhood T cell acute lymphoblastic leukaemia and T cell non‐Hodgkin lymphoma

Mutation analysis of FBXW7 and NOTCH1 genes was performed in 55 T cell acute lymphoblastic leukaemia (T-ALL) and 14 T cell non-Hodgkin lymphoma (T-NHL) patients who were treated on the Japan Association of Childhood Leukaemia Study (JACLS) protocols ALL-97 and NHL-98. FBXW7 and/or NOTCH1 mutations were found in 22 (40.0%) of 55 T-ALL and 7 (50.0%) of 14 T-NHL patients. FBXW7 mutations were found in 8 (14.6%) of 55 T-ALL and 3 (21.4%) of 14 T-NHL patients, and NOTCH1 mutations in 17 (30.9%) of 55 T-ALL and 6 (42.9%) of 14 T-NHL patients. Three (5.4%) T-ALL and two (1.4%) T-NHL patients had mutations in both FBXW7 and NOTCH1. FBXW7 mutations included one insertion, one deletion, one deletion/insertion and nine missense mutations. NOTCH1 mutations were detected in the heterodimerization domain (HD) in 15 cases, in the PEST domain in seven cases, and in both the HD and PEST domains in one case. Five-year event-free survival and overall survival for patients with FBXW7 and/or NOTCH1 mutations were 95.5% (95% CI, 71.9-99.4%) and 100% respectively, suggesting that T-ALL patients with FBXW7 and/or NOTCH1 mutation represent a good prognosis compared to those without FBXW7 and/or NOTCH1 mutations (63.6%, P = 0.007 and 78.8%, P = 0.023, respectively).

Clinical outcomes of different regimens for non-Hodgkin's lymphoma with bone marrow involvement: analysis of 148 cases

To investigate the effects of different treatment protocols on non-Hodgkin's lymphoma (NHL) with bone marrow involvement (BMI). The clinical data of 148 patients of NHL with BMI, 108 undergoing CHOP regimen, 51 undergoing dose-intensive CHOP regimens, 13 undergoing CHOP regimen + rituximab, and 27 undergoing hemopoietic stem cell transplantation (HSCT) after CHOP protocol with remission. The curative effects were retrospectively analyzed. (1) The response rate (RR) for the whole group was 80.4%, with a complete remission (CR) rate of 60.7%. The response rate of the patients treated with the dose-intensive regimens (dose-intensive group) was 84.3%, significantly higher than that of the patients treated with standard CHOP regimen (64.1%, P < 0.05). (2) The 3- and 5-year overall survival (OS) rates and progress-free survival (PFS) of the B-cell NHL patients who underwent HSCT + Rituximab treatment (Rituximab combined with chemotherapy) were significantly higher than those of the dose-intensive group and CHOP group, and the 3- and 5-year OS rates and PFS of the dose-intensive group were all significantly higher than those of the CHOP group (all P < 0.05). (3) The 3- and 5-year OS rates, and PFS of the T-cell NHL patients who underwent HSCT group and dose-intensive CHOP were all significantly higher than those patients who underwent CHOP regimen (all P < 0.05). Superior survival may benefit from Dose-intensive regimens and HSCT, may be good choices for the patients with NHLBMI, a group of heterogeneous malignancies with poor prognosis. And combined chemotherapy with Rituximab treatment remarkably raises the effect for CD20(+) B-cell NHLBMI.

CDC4 and NOTCH1 Mutations in Childhood T-Cell Acute Lymphoblastic Leukemia and T Cell Non-Hodgkin's Lymphoma; a Japan Association of Childhood Leukemia Study Group.

Risk-adapted therapy has resulted in a improvement of outcome in childhood acute lymphoblastic leukemia (ALL), however, T-cell ALL (T-ALL) have still relatively poor outcome compared to B-precursor ALL. Although activating mutations of the NOTCH1 gene have been reported in more than half of T-ALL, the prognostic significance of this finding has yet to be determined. CDC4 gene was isolated as a negative regulator of NOTCH1 and mutations were detected in T-ALL cell line and other cancers. We performed mutation analysis of the NOTCH1 and CDC4 gene in 14 T-ALL cell lines, 52 T-ALL and 17 T cell non-Hodgkin's lymphoma (T-NHL) fresh samples. All children with T-ALL and T-NHL were treated on Japan Association of Childhood Leukemia Study (JACLS) ALL-97 and NHL-98 protocol after obtaining informed consent. Mutation detection was performed via PCR based denaturing HPLC followed by direct sequence.Mutations of the NOTCH1 were identified in 10 (71.4%) of 14 T-ALL cell lines, 16 (30.8%) of 52 T-ALL and 6 (35.3%) of 17 T-NHL fresh samples. Twelve mutations in heterodimerization domain (HD) and 12 mutations in PEST domain (PD) were found in 69 fresh samples. The incidence of the NOTCH1 mutation is less frequent than that of previous reports. We found CDC4 mutations in 12 (35.3%) of 52 T-ALL and 2 (11.8%) of 17 T-NHL fresh samples. One insertion mutation in exon 3 and 11 missense mutations were detected in CDC4 gene. Both NOTCH1 and CDC4 mutations were found in 7 patients. Interestingly, the NOTCH1 mutations were only observed in T-ALL and T-NHL patients without relapse, suggesting to be associated with favorable prognosis. However, CDC4 gene was found not to be associated with prognosis.

Interleukin‐10 gene polymorphism influences the prognosis of T‐cell non‐Hodgkin lymphomas

Interleukin-10 (IL-10) is one of the cytokines implicated in the pathogenesis of diffuse large B-cell lymphoma (DLBCL) in which it acts as auto/paracrine growth factor for lymphoma growth. T-cell non-Hodgkin lymphoma (NHL) is a heterogeneous disease, the biological basis of which is not fully understood. Some evidence suggests that IL-10 might be associated with the progression of T-cell NHLs and that IL-10 may be involved in a rescue effect, protecting T cells from apoptotic cell death associated with upregulated bcl-2 expression. The current study evaluated the impact of IL-10 gene (IL10) polymorphism on the response to chemotherapy and survival in T-cell NHL. IL10 polymorphisms were determined in 108 patients with T-cell NHL. The response to chemotherapy was not dependent on IL10 polymorphism, while survival differed significantly according to IL10 polymorphism. The group with ATA haplotype showed superior overall survival (61.2 +/- 5.9% vs. 21.2 +/- 11.7%, P = 0.001) and failure-free survival (35.0 +/- 5.7% vs. 13.2 +/- 8.7%, P = 0.001) compared to those without ATA haplotype. The ATA haplotype was identified as a favourable prognostic factor compared to non-ATA haplotype (P = 0.037, hazard ratio 2.1), together with international prognostic index (IPI) in a multivariate model for overall survival. In conclusion, IL10 polymorphism may affect the survival of T-cell NHL patients.

The clinical value of follow‐up examinations in childhood T‐cell acute lymphoblastic leukemia and T‐cell non‐Hodgkin's lymphoma

The aim of this study was to evaluate the value of follow-up investigations of T-cell acute lymphoblastic leukemia (T-ALL) and T-cell non-Hodgkin's lymphoma (T-NHL), including cerebrospinal fluid (CSF) examination, bone marrow (BM) aspiration, peripheral blood (PB) count, serum lactate dehydrogenase (LDH) and chest X-rays in patients with an initial mediastinal enlargement. We reviewed clinical records of all T-ALL patients from 1987 to 2002 and all T-NHL patients from 1977 to 2002, seen at a single institution. Of 48 T-ALL patients, 15 suffered from a relapse, 6 (40%) were asymptomatic at the time of relapse. T-ALL (13/30) with mediastinal enlargement at first diagnosis relapsed versus 2/16 of those without mediastinal enlargement. However, at relapse, only one patient had a mediastinal mass, which in addition was symptomatic. Of 39 T-NHL patients, 6 patients relapsed. Forty percent of relapsed T-ALL and 17% of relapsed T-NHL were asymptomatic. The seven asymptomatic relapses were detected by CSF (n = 4), BM (n = 2) or blood count (n = 1) examinations. All T-ALL and T-NHL patients with a mediastinal relapse were symptomatic. This study suggests that routine CSF examinations during treatment can detect relapses of T-ALL and T-NHL before onset of symptoms, which might be of clinical value. Relapses are rarely detected by BM or blood examinations and whether this translates in a clinical benefit is unlikely. Routine chest X-rays are not useful.

Allogeneic Hematopoietic Stem Cell Transplantation in T-Cell Non-Hodgkin Lymphoma.

T-cell non-Hodgkin Lymphoma (T-NHL) are uncommon malignancies that represent approximately 10% of all NHL. With few exceptions T-NHL have generally been reported to have a worse prognosis compared with B-cell lymphomas. Standard therapy for most T-NHL still need to be defined. To elucidate the feasibility and efficacy, we retrospectively analysed the data of 11 patients who underwent allogeneic hematopoietic stem cell transplantation from July 1991 to july 2006. Five female and five male patients with a matched sibling donor and a female mismatched underwent a transplant. The median age was 22 years (16 – 34). Six patients were diagnosed as lymphoblastic T, two were anaplastic large cell, two were peripheral T-cell unspecified and one was NK/T extranodal nasal-type. One was in first hard remission, three in second remission, one in third complete remission, one in early first relapse, one in second partial remission and four had refractory disease. Six patients were conditioned with TBI + cyclophosphamide (CY-TBI) and five with busulfan 16 mg/kg and melphalan 140 mg/sqm (Bu-Mel). Graft versus host disease (GVHD) prophylaxis was done with cyclosporin and short course methotrexate for most of the cases. 63 % had acute GVHD grade I to III and 45% had chronic GVHD. Overall survival is 36%. Four of the 11 patients are alive, still in complete remission, after one year, 12 years, 14 years, and 15 years from transplantation. Two of these patients were conditioned with CY-TBI and two with Bu-Mel. Allogeneic transplant can be useful to treat advanced T-NHL patients.

Prospective analysis of treatment outcome and prognostic factors in patients with T‐cell lymphomas treated by CEOP‐B: single institutional study

The important prognostic factors were evaluated for T-cell non-Hodgkin lymphoma (NHL) patients in a prospective study using the CEOP-B protocol [a modified cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP)-like regimen that uses epirubicin instead of doxorubicin with the addition of bleomycin]. Fifty-two patients were enrolled in the study. The overall response rate was 63.5%. The median progression-free survival (PFS) and median overall survival (OS) was 18.0 and 39.5 months respectively. The most common toxicity was neutropenia. The factors related to poor outcome were a high International Prognostic Index (IPI) and a high 'B' score (bone marrow involvement, B symptoms, bulky disease). We developed a new prognostic model, namely the Prognostic Group for T cell NHL (PGT) that included four groups: PGT1 (low IPI/low B score), PGT2 (low IPI/high B score), PGT3 (high IPI/Low B score) and PGT4 (high IPI/Low B score). OS and PFS (not reached, 48 months) in the PGT1 group were significantly longer than those (11.5 and 4.8 months) in PGT2. The same result was observed in the PGT3 and PGT4 groups. The CEOP-B regimen was moderately active and tolerable for T-cell NHL patients, and the PGT system might be useful for the prediction of long-term survival of T-cell NHL patients.