Mantle Cell Lymphoma Group Research Articles

Central nervous system involvement by mantle cell lymphoma

Involvement of the central nervous system (CNS) is a rare complication of mantle cell lymphoma (MCL) with limited treatment options. We report the outcomes of 36 patients with CNS involvement compared to 72 matched control MCL patients without CNS involvement. Four patients (11%) with CNS MCL were diagnosed with CNS involvement at time of MCL diagnosis. Median OS from MCL diagnosis was 50.3 months (95% CI: 22.8–79.6) for the CNS MCL group compared to 97.1 months (95% CI: 82.8–NR; p= <0.001) for the control group. Median OS from CNS involvement was 4.7 months (95% CI: 2.3–6.7). CNS involvement by MCL has dismal outcomes as evident by a short median OS and PFS after CNS involvement. Advanced stage, blastoid variant, elevated LDH, and elevated Ki67 at MCL diagnosis were features more commonly seen in the CNS MCL cohort.

Central Nervous System Involvement By Mantle Cell Lymphoma

Central Nervous System Involvement By Mantle Cell Lymphoma

Dilemma-High Response Rates Vs High Toxicity: Autologous Stem Cell Transplantation for Lymphoma

Introduction: Autologous stem cell transplantation (ASCT) is employed successfully at different settings in lymphoma. Most of the patients have been achieving durable remission rates with long term survival. Herein, our aim is to report outcomes of our relapse refractory Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) patients who underwent ASCT.Methods: We retrospectively evaluated 86 relapse refractory HL, 21 mantle cell lymphoma, 16 low grade lymphoma (n=12 follicular lymphoma, n=4 marginal zone lymphoma) patients who underwent ASCT between 2005-2015. The patients were followed after ASCT for relapse.Results: The baseline characteristics of patients are given in the table. The patients were diagnosed as nodular sclerosing subtype most frequently in HL group (75%). In HL group, 27 patients (32%) had chemo-refractory high risk disease prior to ASCT. On the other hand, all patients in mantle cell and low grade lymphoma groups were in at least partial remission at the time of ASCT. In HL group, at a median time of 28 months after ASCT (range, 1-119), 51 patients (60%) achieved complete remission, 7 patients had partial remission (8%), 28 patients (32%) had stabile disease or progression/relapse. The response was higher in patients with early stage disease (I-II), remission status prior to transplant and BEAM conditioning regimen compared to advance stage (III-IV) (p=0.017), active disease prior to transplant (p=0.008) and other conditioning regimens (p=0.05). Nine patients (10%) died within three months of ASCT due to infection. From 28 patients, 5 patients received brentixumab, 3 patients received nivolumab, 10 patients had GDP, 10 patients had MOPP regimen. Four patient underwent allogeneic stem cell transplantation. Fifty four out of 58 (93%) patients transplanted with responsive disease were alive without disease whereas 4 out of 28 (14%) patients with unresponsive disease were alive in remission within 3 years post ASCT. In low grade lymphoma group, 9 patients (56%) achieved complete remission after ASCT. Interestingly, complete response rate was higher in females (80% vs 43%, p=0.35). Five patients (31%) died within three months of ASCT due to infection. Two patients (13%) relapsed with a median time of 8 months (range, 3-13 months) and received MINE as salvage treatment and followed in complete remission. The 3-year overall survival was 51%, 5-year overall survival was 40%. In mantle cell lymphoma group, 6 patients (29%) achieved complete remission. Four patients (19%) died within three months of ASCT due to infection. Eleven patients (52%) was relapsed with a median time of 39 months (range, 4-123 months). Ten patients received BORID (bortezomib, rituximab, dexamethasone) and 1 patient received lenalidomide as salvage therapy and six of them achieved complete remission. Three patients underwent allogeneic hematopoietic stem cell transplantation as well as two patients received ibrutinib as fourth line therapy and followed in remission. The 3-year overall survival was 71% (Figure).Conclusion: In our experience, ASCT is a feasible option in patients who responded showing long term durable complete remission rates. In Hodgkin lymphoma, patients with BEAM conditioning, remission status prior to ASCT and early stage disease had better response rates. Females achieved higher responses in low grade lymphomas. However, 10% of HL, 31% of low grade lymphoma and 19% of mantle cell lymphoma patients were died because of treatment related toxicity. Candidate patients should be evaluated carefully. Today, novel agents may present a less intensive approach for achieving response. [Display omitted] DisclosuresIlhan:Novartis: Consultancy.

A Phase II Trial Evaluating the Efficacy of Radioiodinated Tositumomab (Anti-CD20) Antibody, Etoposide and Cyclophosphamide Followed By Autologous Transplantation, for High-Risk Relapsed or Refractory Non-Hodgkin's Lymphoma

BACKGROUND: High dose therapy followed by autologous stem cell transplantation (ASCT) is commonly used for treating patients with high-risk, relapsed or refractory non-Hodgkin's lymphoma (NHL). Total body irradiation (TBI) has historically been used as a part of the conditioning component based on the exquisite sensitivity of lymphoma to radiation. However, the dose of radiation delivered is limited to major organ toxicity. One strategy to enhance the absorbed dose to target sites and decrease the dose to major organs is to replace external beam TBI with high-dose radioimmunotherapy (RIT). Here we describe the first report of a phase II trial evaluating high-dose I-131-tositumomab (anti-CD20), etoposide (VP-16) and cyclophosphamide (CY) followed by ASCT for high-risk, relapsed or refractory NHL.METHODS: A single arm, Phase II study was carried out at The Fred Hutchinson Cancer Research Center and University of Washington Medical Center. Patients age < 60 years with histologically confirmed lymphoma expressing the CD20 antigen, with tumor burden < 500 cc, without splenomegaly, acceptable organ function (Cr < 2.0, bilirubin < 1.5mg/dL, normal cardiac ejection fraction, DLCO ≥ 50%), and adequate number of cryopreserved autologous PBSC were eligible. Patients must have had relapsed or refractory disease, except mantle cell lymphoma (MCL) that could be transplanted in first remission. Patients received trace labeled (~10mCi) I-131-tositumomab followed by serial quantitative gamma camera imaging to calculate organ doses and determine the I-131 activity needed to deliver ≤ 25 Gy to critical normal organs. Patients received individualized doses of therapeutic I-131-tositumomab to deliver 25 Gy to the dose limiting normal organ on ~ Day -14, VP-16 60mg/kg on Day -4, CY 100mg/kg on Day -2, followed by ASCT on Day 0. The primary endpoint was progression free survival (PFS) with secondary endpoints of overall survival (OS), engraftment, and non-relapse mortality (NRM).RESULTS: A total of 108 patients were enrolled, treated, and stratified into one of three groups: aggressive lymphoma (n = 29), indolent lymphoma (n = 46), and MCL (n = 33). The median age was 52 years (range 32-60), 69% were male, median prior regimens was 3 (range 1-11), 60% had chemotherapy responsive disease at time of ASCT and 91% had prior Rituximab (R) including 26 of 29 patients with aggressive B-NHL who had received a prior R-chemotherapy based regimen, and 32% had R-refractory disease including 17 of 46 patients with indolent B-NHL. Table 1 shows histologic subtypes at enrollment. Patients received a median of 557 mCi of I-131 (range 231-1353). The most common dose-limiting organs were lung (n = 70), liver (n = 27), and kidneys (n= 6). The median time to engraftment of ANC > 500 and Plt > 20K was 12 and 12 days respectively. The 30-day NRM was 0%, and the 100-day NRM was 2.8%. With a median follow up of 10.1 yrs, the 5 and 10 year PFS for the aggressive, indolent, and MCL groups were 55%, 62%, 61% and 46%, 56%, 42% respectively (figure 1), meeting or exceeding pre-specified phase II benchmarks for this protocol. The 5 and 10 year OS for the aggressive, indolent, and MCL groups were 69%, 80%, 67% and 61%, 70%, 48% respectively. In patients with aggressive B-NHL, prior R-chemotherapy was not associated with inferior PFS (p = 0.7). Likewise, PFS outcomes were similar for R-refractory or R-sensitive indolent B-NHL (p = 0.8). Late myeloid malignancies were seen in 6 (5.6%) patients.CONCLUSIONS: High-dose I-131-tositumomab (anti-CD20) antibody, VP-16 and CY followed by ASCT appears feasible, safe, and effective in treating B-NHL, resulting in an estimated 42-56% of patients being alive and progression-free at 10 years. Short and long-term toxicity appear comparable to standard conditioning regimens. RIT-containing transplant regimens should thus be evaluated in a prospective randomized trial. [Display omitted] DisclosuresMartin:Procter and Gamble: Equity Ownership; Pfizer: Consultancy; Fresenius, Neovii: Research Funding; Incyte: Consultancy. Holmberg:Seattle Genetics: Research Funding; Sanofi: Research Funding; Merck: Research Funding; Millennium: Research Funding; JAZZ: Consultancy; Up To Date: Patents & Royalties. Pagel:Pharmacyclics: Consultancy; Gilead: Consultancy.

Primary Analysis of the Tegar Study: A Phase II Study Exploring the Safety and Efficacy of Atezolizumab in Combination with Obinutuzumab or Rituximab in Patients with Relapsed or Refractory (R/R) Mantle Cell Lymphoma (MCL), Marginal Zone Lymphoma (MZL), or Waldenstrom Macroglobulinemia (WM)

Primary Analysis of the Tegar Study: A Phase II Study Exploring the Safety and Efficacy of Atezolizumab in Combination with Obinutuzumab or Rituximab in Patients with Relapsed or Refractory (R/R) Mantle Cell Lymphoma (MCL), Marginal Zone Lymphoma (MZL), or Waldenstrom Macroglobulinemia (WM)

Efficacy and Safety Analysis of Ibrutinib-Containing Therapy for Relapsed/Refractory (R/R) Mantle Cell Lymphoma (MCL): Results from a Real-World Study in China

Background: Ibrutinib, a first-in-class Bruton's tyrosine kinase (BTK) inhibitor, has been approved for the treatment of relapsed/refractory (R/R) Mantle cell lymphoma (MCL). Both single-agent ibrutinib and combination of ibrutinib with rituximab have achieved great efficacy with manageable toxicity (Wang,NEJM2013; Wang,Lancet Oncol2015; Dreyling,Lancet2015). This study, for the first time, analyzed the real-world effectiveness and tolerability of ibrutinib for MCL patients in China. Methods: This multi-center, retrospective cohort study enrolled adult patients (pts) with pathologically confirmed MCL who initiated ibrutinib-containing treatment between November 2017 (date of commercialization) and April 2020. Eligible patients were retrospectively divided into 3 subgroups to receive ibrutinib-containing treatments for different purposes: R/R MCL group, newly diagnosed MCL group and maintenance therapy group. This analysis reports the baseline characteristics, efficacy and safety profiles in R/R MCL patients. Results: A total of 67 R/R MCL pts receiving ibrutinib-containing treatment from 9 medical centers in China were included in this analysis. At ibrutinib initiation, the median age was 61.0 (range 39-81) years, 68.7% were male and 81.8% had Eastern Cooperative Oncology Group (ECOG) performance status of 0-1. Approximately three-quarters of pts (72.3%) had 1 previous line of therapy before ibrutinib. Baseline characteristics are summarized in Table 1. Of all patients enrolled, 53.7% (36/67) of pts received ibrutinib monotherapy and 46.3% (31/67) received ibrutinib-containing combination therapy. IR (ibrutinib and rituximab) (16/31, 51.6%) and IR2 (ibrutinib, rituximab and lenalidomide) (5/31, 16.1%) were the two most common combination regimens. Nine patients (29.0%) received ibrutinib plus R-chemotherapy. Although no statistically significant difference was found in listed baseline characteristics between these two groups, a larger percentage of pts with bone marrow involvement (58.1% vs 35.7%) and bulky mass (largest diameter) ≥5 cm (46.4% vs 27.3%) were observed in combination therapy group. Best overall response rate (ORR) was 65.7% (20.9% complete remission [CR]). Median time to response (TTR) was 4.1 months and median duration of response (DOR) was 18.4 months. With a median follow-up of 10.2 months, median progression-free survival (PFS) was 21.3 months (95% confidence interval [CI], 15.2 - not available [NA]) (Figure 1A). PFS rates were 86.0%, 69.8% and 47.6% at 6 months, 1 year and 2 years. With a median follow-up of 11.2 months, median overall survival (OS) was not reached with OS rates of 98.5%, 87.9% and 76.3% at 6 months, 1 year and 2 years (Figure 1B). Compared with ibrutinib monotherapy, combination therapy showed higher ORR (50.0% vs 83.9%), CR rate (8.3% vs 35.5%) and shorter TTR (median TTR, 6.0 vs 2.2 months; Logrank,p=0.0012) (Figure 2A). Although the combination therapy had a trend for better PFS, no statistically significant benefit in PFS or OS was observed (Figure 2B, C). Safety analysis focused on 55 R/R MCL pts from 3 centers with adequate adverse events information. The most common treatment emergent adverse events (TEAEs) of interest were infection (8/55, 14.6%), rash (8/55, 14.6%), bleeding (5/55, 9.1%; 1/5 was major bleeding [subdural hemorrhage]) and atrial fibrillation (3/55, 5.5%). Four pts (7.3%) experienced grade 3-4 TEAE (neutropenia, n=2; neutropenia and lung infection, n=1; subdural hemorrhage, n=1). Six pts (10.9%) had ≥1 temporary ibrutinib discontinuation due to TEAE (infection, n=3; neutropenia, n=1; rash, n=1; vomiting, n=1). One patient discontinued ibrutinib permanently due to TEAE (subdural hemorrhage). Combination therapy group showed a higher incidence of hematological TEAE (60.9% vs 39.1%) and infection (20.7% vs 7.7%). No TEAE-related death or new safety signals was recorded. Conclusion: This real-world analysis demonstrates that ibrutinib is effective and tolerable for R/R MCL in China. Ibrutinib-containing combination therapy outperformed ibrutinib monotherapy in response rate and TTR, but showed no survival benefits. The response rate data of ibrutinib monotherapy obtained from our study was different to existing clinical trial data, which may be mainly due to the short follow-up time of our study. Further analysis with longer follow-up is needed to validate these findings. Disclosures No relevant conflicts of interest to declare.

Relationship of Expression Between SOX11 and PAX5 in Pathological Tissue Specimens of Patients with Mantle Cell Lymphoma and Its Clinical Significance

Investigate the expression of SRY-related HMG box 11 (SOX11) and paired box domain 5 (PAX5) in patients with mantle cell lymphoma (MCL) and analyze the relationship between them and their clinical significance. Seventy-six formalin-fixed paraffin-embedded (FFPE) samples of patients who were diagnosed with MCL from January 2012 to August 2017 were collected．Fifty-six FFPE samples from patients with diffuse large B cell lymphoma (DLBCL), thirty-eight FFPE samples from patients with follicular lymphoma (FL) and nine FFPE samples from patients with Burkitt's lymphoma (BL) were used as control groups. Real-time quantitative PCR (qRT-PCR) and immunohistochemistry were used to detect the mRNA and protein expressions of SOX11 and PAX5. The association between expressions of SOX11 and PAX5 in patients with MCL was analyzed. On the basis of the median H score of SOX11 and PAX5 protein expressions in patients with MCL, they were divided into high and low expression group, and the relationship between the different groups and patients' clinical characteristics and prognosis were analyzed. The different mRNA expression levels of SOX11 and PAX5 in different lymphoma tissues were statistically significant ( P<0.01). The mRNA expression levels of SOX11 and PAX5 in MCL group were higher than those of the control groups, and the differences of those between MCL and DLBCL or FL were statistically significant ( P<0.01). However, the differences of those between MCL and BL were not significant ( P>0.05). The expression level of SOX11 protein was also higher than those of the control groups ( P<0.000 1). However, there was no significant difference in PAX5 protein expression level between the MCL group and the control group, nor the expression levels of SOX11 and PAX5 genes and proteins among the control groups ( P>0.05). By analyzing the samples from patients with MCL, we observed a positive relevance between SOX11 and PAX5 both in mRNA expression level ( r s=0.714, P<0.000 1) and protein expression level ( G=0.407, P=0.01). There was no difference in clinical characteristics and overall survival between the high and low expression group. In MCL, there was a positive relevance between the expressions of SOX11 and PAX5. The expression of SOX11 or PAX5 alone has no significant effect on the prognostic stratification of MCL patients.

Ibrutinib Monotherapy in Patients with Relapsed/Refractory Mantle Cell Lymphoma in Community Clinical Practice Outside of Clinical Trials

Background. Ibrutinib became available for patients (pts) with relapsed/refractory (R/R) mantle cell lymphoma (MCL) in Russia in the end of 2015 based on the results of phase 2 study by M. Wang from MD Anderson Cancer Center (NEJM 2013, Blood 2015). However, pts in routine clinical practice tend to differ significantly from the patients selected for clinical trials, which may lead to poorer results in off study treated patients. Aim. To assess efficacy and toxicity of ibrutinib monotherapy in pts with R/R MCL in routine community clinical practice outside of clinical trials. Materials &amp; Methods. We analyzed the charts of all pts with R/R MCL who started ibrutinib from April 2016 to July 2019 in 7 Moscow's hospitals. The following criteria were used to initiate ibrutinib monotherapy: the age &gt; 18 years, confirmed MCL diagnosis with nuclear hyperexpression of cyclin D1 and/or presence of the t(11;14)(q13;q32); symptomatic relapse or failure to achieve at least PR with a prior regimen. Poor physical status, pancytopenia grade 3-4, infectious complications (except for life-threatening conditions), blastoid variant and the number of previous treatment lines were not regarded as contraindications to ibrutinib therapy. Patients with CNS involvement were excluded from this analysis. Ibrutinib was administered once a day at a dose of 560 mg until progression or intolerable toxicity. Response to therapy was assessed every 2-3 months utilizing CT scan, PET/CT and bone marrow examination were required to confirm CR. Results. 54 pts with R/R MCL received ibrutinib monotherapy between April 2016 and July 2019. 26 pts (48%) were refractory to a prior therapy. The median age was 68 years (range 40-81); 69% of pts were men; ECOG &gt; 2 in 22% of pts. 15 pts (28%) underwent a repeated biopsy before starting ibrutinib. 23 pts (43%) had an aggressive variant of MCL: either blastoid morphology (13/54 at diagnosis and 5/54 after repeated biopsy) or classical morphology with Ki-67&gt;40% (4/54 at diagnosis and 1/54 after repeated biopsy). The median number of previous treatment lines was 2 (1-11). The response was evaluated in 53/54 pts. ORR was 81%, CR rate was 30% in the whole group. Pts with aggressive variants of MCL (group 1) had ORR of 65%, CR rate of 7%. In the group with classical morphology and Ki-67≤40% (group 2) ORR and CR rates were 93% and 47%. 2 pts with a response to therapy stopped ibrutinib early on their own accord, without signs of toxicity above gr.1. The median EFS for all pts was 394 days (95% CI: 261-526). In group 1 the median EFS was 173 days (95% CI: 112-234), in the group 2 the median EFS was 759 days (95% CI: 521-996, p&lt;0.0001). 27 pts (50%) died (25 deaths were MCL-related). The median OS for the entire population studied was 491 days (95% CI: 183-799). The median OS of pts after progression on ibrutinib was disappointingly short - 84 days (95% CI: 30-139). 21 pts (39%) remain on ibrutinib treatment for 10-1119 days (the median duration was 358 days). The most common complications were myalgia and muscle cramps (60%, all gr.1-2), diarrhea (gr.1-2 in 58% and gr.3 in 4%); hemorrhagic complications (65%, all gr. 1-2); skin, nail toxicity and a rash (15%, all gr.1-2) and arrhythmia (9%). Infections were reported in 30% of pts (gr.3 in 6%). Skin/nails toxicity developed mainly after 6 months of ibrutinib intake. In 14 (26 %) pts ibrutinib treatment had to be adjusted (dose reduction or treatment interruption for 3-10 days) due to toxicity and planned surgeries. 4 pts (9%) required dose reduction from 560 to 420 mg. None of ibrutinib recipients had to completely discontinue ibrutinib therapy due to complications (with exception of 2 patients who stopped taking ibrutinib on their own). None of the patients received alloSCT. Conclusion. These data on the use of ibrutinib in actual clinical practice are comparable with the results of international multicenter studies (PCYC-1104, SPARK, and RAY). Favorable toxicity profile and rather short time to antitumor response allow for ibrutinib administration in cases of poor performance status due to disease, low blood count, and even infectious complications. Ibrutinib monotherapy results in the durable disease control in 93% of patients with non-blastoid MCL with the median EFS 25 months, but only 65% with blastoid type with the median EFS 6 months. However, some adverse effects are manifested not earlier than after 6-month treatment, which calls for continuous monitoring, especially when preparing for surgeries. Figure Disclosures Vorobyev: AstraZeneca: Consultancy; Abbvie: Consultancy, Speakers Bureau; Roche: Consultancy, Speakers Bureau; Janssen: Consultancy, Speakers Bureau; Takeda: Consultancy, Speakers Bureau; Sanofi: Consultancy. Ptushkin:JSC GENERIUM: Research Funding; Alexion Pharmaceuticals, Inc:: Consultancy, Research Funding; Janssen: Consultancy; Roche: Consultancy; AbbVie: Consultancy.

Expression of MiR101 and EZH2 in Patients with Mantle Cell Lymphoma and Its Clinical Significance

To investigate the expression of miR-101 and EZH2 in patients with mantle cell lymphoma(MCL) and to analyze its correlation with clinical prognosis of MCL patients. RQ-PCR and S-P immunohistochemistry were used to detect the expressions of miR-101 and EZH2 in tissue of MCL patients. CCK-8 was used to assay the effect of miR-100 minics on the proliferation of Jeko-1 and Mino cells; the flow cytometry with Annexin V/PI double staining was used to assay the apoptosis; Western blot was used to assay the effect of miR-101 minics on the expression of EZH2 protein in Jeko-1 and Mino cells. Compared with control group, miR-101 lowly expressed, and EZH2 protein highly expressed in MCL group, with very statistically significant difference(P<0.01).There was negative correlation between miR-101 and EZH2 expression(r=-0.638，P<0.05). The expression of miR-101 and EZH2 significantly correlated with B symptoms, International Prognostic Index(IPI) and Ann Arbor stage, respectively. Survival analysis showed that the overall survival(OS) rate of patients with low expression of miR-101 were significantly lower than that of patients with high miR-101 expression (P=0.0014), the OS rate of patients with EZH2 high expression were significantly lower than that of patients with EZH2 low expression (P=0.0093). The miR-100 minics could inhibit the proliferation of Jeko-1 and Mino cells, and increase the apoptotic rate. The expression of EZH2 protein was markedly suppressed by the miR-100 minics. The expression of miR-101 and EZH2 is different in MCL patients with different clinical stage and prognosis. The miR-101 can inhibit the cell proliferation and induce cell apoptosis of mantle cell lymphoma by targeting EZH2.

CD5 Negative Mantle Cell Lymphoma: Clinicopathologic Correlations and Outcome in 58 Cases

Introduction: Mantle cell lymphoma (MCL) is a B-cell neoplasm that has a characteristic immunophenotype of being positive for CD5, B-cell antigens and cyclin D1. A small subset of cases of MCL can be negative for CD5, approximately 5% in the literature. The clinicopathologic features and prognosis of patients with CD5-negative MCL are poorly characterized. Here, we study a group of patients with CD5- MCL and compare them with a group patients with CD5+ MCL.Methods: From a total of 270 cases of MCL accessioned from 2004-2015, 58 CD5- cases (study group) and 212 CD5+ cases (control group) were identified. All cases of MCL were positive for cyclin D1 by immunohistochemistry and, in most patients, CCND1-IGH was shown FISH. Cases negative for CD5 were assessed by flow cytometry and/or immunohistochemistry. Fisher exact test was utilized to analyze differences between the CD5- and CD5+ groups. Patient survival was analyzed using the Kaplan-Meier method and compared using the log-rank test. Univariate and multivariate Cox proportional hazards model analyses for OS and PFS were performed (SPSS 22 software). A P-value of less than 0.05 was considered statistically significant.Results: The CD5- group included 39 men and 19 women with a median age of 66 years (range, 36- 88 years) at time of diagnosis. The CD5- and CD5+ groups shared overlapping clinicopathological features, but CD5- cases showed a lower percentage of men (P=0.006) than CD5+ cases. Treatment information was available for 50 patients. Twenty-nine (58%) patients were treated initially with R-Hyper CVAD therapy (rituximab, fractionated cyclophosphamide, vincristine, doxorubicin, and dexamethasone alternating with high dose methotrexate and cytarabine). Seventeen (34%) patients were treated initially with less aggressive therapy: 7 with R-CHOP; 8 had other rituximab-based chemotherapy regimens; 2 received rituximab as a single agent. Four patients (8%) were observed without therapy. After induction, 34 patients achieved complete remission (CR), 5 patients achieved partial remission (PR), 6 patients showed no response (NR) or progressive disease (PD), and 5 patients lost follow-up. Ten patients also underwent stem cell transplantation (SCT): 5 patients received allogeneic SCT, the other 5 autologous SCT. With a median follow-up of 45.7 months (range, 2.0-174.3 months), 13 of 56 (23.2%) patients died, 43 of 56 (76.8%) patients were alive at last follow-up, and the rest of 2 patients lost follow up. The induction chemotherapy regimens and CR and PR rate were not significantly different between the CD5- and CD5+ groups (p>0.05). Survival analysis showed patients with CD5- MCL had a tendency for longer OS (Figure 1A, P=0.078). Further analysis showed that lack of CD5 expression predicted a superior OS in a few subsets of MCL patients defined with 1) normal WBC count (p=0.049); 2) Stage I/II disease (p=0.046); 3) Low/intermediate MIPI (p=0.041) and 4) Ki67≥30% (at a borderline p value of 0.05). Patients with CD5- MCL also showed a significantly longer progression-free survival (PFS) (Figure 1B, P=0.01). Absence of CD5 expression was associated with a better PFS in MCL patients with advanced disease (stage III-IV) (P=0.035), a normal leukocyte count (P=0.018), a normal serum lactate dehydrogenase level (P=0.046), classical morphology (P=0.029), and low/intermediate MIPI (p=0.0004). Multivariate Cox regression analysis revealed that MIPI was the only independent prognostic factor for both OS and PFS (P=0.026 and P=0.001 respectively) and CR/PR also predict a better OS (P=0.004) in CD5- MCL patients.Conclusion: The clinicopathologic features were similar between patients with CD5- MCL and those with CD5+ MCL, except that less men in the CD5- MCL group. Lack of CD5 expression was associated with a favorable PFS in MCL patients. Recognizing this subgroup of CD5- MCL has not only a diagnostic significance, but also a prognostic significance. [Display omitted] DisclosuresWang:Acerta Pharma: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Juno Therapeutics: Research Funding; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Kite Pharma: Research Funding; Asana BioSciences: Research Funding; BeiGene: Research Funding; Pharmacyclics: Research Funding; Celgene: Research Funding; Onyx: Research Funding.

Platelet Function Prior to Ibrutinib Treatment Is More Severely Impaired in Chronic Lymphocytic Leukemia Than in Mantle Cell Lymphoma

INTRODUCTION. Ibrutinib is an oral small-molecule drug that irreversibly inhibits Bruton's tyrosine kinase and thus suppresses proliferation of B cells. Ibrutinib therapy in chronic lymphocytic leukemia (CLL) and in mantle cell lymphoma (MCL) is associated with frequent bleeding complications that were reported to be more severe in CLL. This difference was suggested to be related to differential adenosine diphosphate (ADP) disruption by CD39, expressed by tumor cells. Here we investigate platelet functional activity in CLL and MCL patients before initiation of ibrutinib.MATERIALS. Fifty one adult patients with relapsed and refractory CLL and MCL and 20 healthy donors were included in the study. Platelet functional activity was characterized by flow cytometry before and after activation with SFLLRN plus collagen-related peptide. Levels of CD42b, CD61, CD62P, PAC1, annexin V binding, and mepacrine release were determined. Aggregation with collagen, ADP and ristocetin were measured.RESULTS. Among 36 CLL patients 25 (69%) were men, the median age was 66 (range, 31 to 83 years) and 23 (64%) had a refractory disease. In 34 previously treated patients the median number of prior treatments was 2 (range, 1 - 6). Two CLL patients with del (17p) received ibrutinib as a first line. In a group of MCL patients (n=15) there were 12 men (80%), the median age was 63 years (range, 49-79). Seven (54%) patients had a high MIPI score. The median number of previous chemotherapy lines in MCL patients was 1 (range, 1 - 6). Lymphocyte count before initiation of ibrutinib was significantly higher in CLL group (median 17,5 х 109/L range 0,48 - 189,7) compared to MCL group (median 1,75 х 109/L, range 0,35 - 6,59, p=0,037). Initial platelets count was also different being lower in CLL than in MCL patients (127 х 109/L (15 - 248) versus 186 х 109/L (63 - 311), p=0,005). Platelets of untreated patients with CLL had statistically significant impairments in their responses compared to those with MCL. Specifically, they had decreased integrin activation in response to stimulation (46±19% versus 73±18%; for healthy donors, 100±17%), and relative mepacrine release (2.9±0.9 versus 3.5±0.8; healthy, 4.0±0.7). P-selectin release was impaired milder (77±17% versus 92±14%; healthy, 100±10%), and procogulant platelet formation impairment was not statistically significant (8±6% versus 12±7%; healthy, 12±3%). In control experiments, addition of MeS-AMP and MRS2179, selective inhibitors of ADP receptors, had comparable effects on the healthy donor platelets. Aggregation in response to ADP was dramatically impaired in CLL compared with MCL (24±18% versus 42±16%), as well as that in response to collagen (47±23% versus 72±13%). Upon ibrutinib treatment, further decrease of PAC1 binding and procoagulant platelets formation almost to the level of resting platelets were observed; activation with SFLLRN alone produced a similar result with donor platelets.CONCLUSIONS. Patients with CLL and MCL have initially impaired response of platelets. The defects of platelet function in CLL is much greater suggesting a possible explanation of the different clinical severity. DisclosuresVorobyev:Janssen: Speakers Bureau. Ptushkin:Janssen: Speakers Bureau.

Pathologic characteristics of bone marrow for CD5 positive small B cell lymphoma

To study the pathologic characteristics of bone marrow for CD5 positive small B cell lymphoma (SBL). The pathologic profiles of 92 patients with CD5 positive SBL were retrospectively analyzed. The morphologic and immunophenotypic features were analyzed by flow cytometry and immunohistochemistry. IgH/CCND1 was examined by fluorescence in situ hybridization (FISH). A total of 92 patients with CD5 positive SBL were enrolled in this study, including 56 (60.9%) chronic lymphocytic leukemia /small lymphocytic lymphoma (CLL/SLL), 23 (25.0%) mantle cell lymphoma (MCL) and 13 other SBL (14.1%). Among the 13 other cases, 5, 4 and 4 cases were follicular lymphoma (FL), lymphoplasmacytic lymphoma (LPL) and splenic marginal zone lymphoma (SMZL), respectively. The frequency of patterns for bone marrow infiltration was as follow: diffuse pattern (19/92), mixed pattern (15/92), nodular pattern (9/92), interstitial pattern (8/92), and intrasinusodial pattern (2/92). All patients expressed CD19, CD20 and CD5. According to the immunophenotypic score system, all the CLL patients had 4-5 scores, while SMCL and other SBL patients had less than 3 scores. For the other SBL patients, 5 FL expressed CD10, while 3 FL, 1 LPL and 3 SMZL expressed CD23. There was a significant difference in the expression of CD23, sIgM, FMC7, CD11C and CD22 between the CLL and MCL groups (P<0.01). All 23 MCL patients expressed cyclin D1 and showed IgH/CCND1 gene translocation by FISH detection. CD5 positive SBL includes a variety of types of lymphoma. Patterns of bone marrow for CD5 positive SBL are diversity. Immunophenotypic analysis by flow cytometry is essential in the diagnosis and differential diagnosis of CD5 positive SBL, especially for CLL.

Role of CyclinD1/IgH Detection by FISH in Differential Diagnostic Significance between Mantle Cell Lymphoma and Chronic Lymphocytic Leukemia

To investigate the feasibility of CyclinD1/IgH detection by FISH in diferential diagnosis between mantle cell lymphoma (MCL) and chronic lymphocytic leukeamia (CLL). The FISH detection was performed for CyclinD1/IgH fusion gene. A comprehensive analysis was carried out for clinical features, such as age, sex , WBC count and lymphocyte count, the bone marrow morphology and immunohistochemical staining were carried for CyclinD1/IgH. It is often difficult to distinguish MCL from CLL by bone marrow morphology, when the cell morphology was not typical; there was no difference in age, sex, WBC count and lymphocyte count between MCL and CLL groups; 9 out of 52 patients were diagnosed as MCL, and the direction of CyclinD1/IgH by FISH was positive in 7 of 9 MCL, while 3 of the 7 patients were negative by immunohistochemical staining for CyclinD1. Detection of CyclinD1/IgH by FISH can be used as a specific and feasible method for differential diagnosis of mantle cell lymphoma from chronic lymphocytic leukeamia.

Distinction between Asymptomatic Monoclonal B-cell Lymphocytosis with Cyclin D1 Overexpression and Mantle Cell Lymphoma: From Molecular Profiling to Flow Cytometry

According to current diagnostic criteria, mantle cell lymphoma (MCL) encompasses the usual, aggressive variants and rare, nonnodal cases with monoclonal asymptomatic lymphocytosis, cyclin D1-positive (MALD1). We aimed to understand the biology behind this clinical heterogeneity and to identify markers for adequate identification of MALD1 cases. We compared 17 typical MCL cases with a homogeneous group of 13 untreated MALD1 cases (median follow-up, 71 months). We conducted gene expression profiling with functional analysis in five MCL and five MALD1. Results were validated in 12 MCL and 8 MALD1 additional cases by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and in 24 MCL and 13 MALD1 cases by flow cytometry. Classification and regression trees strategy was used to generate an algorithm based on CD38 and CD200 expression by flow cytometry. We found 171 differentially expressed genes with enrichment of neoplastic behavior and cell proliferation signatures in MCL. Conversely, MALD1 was enriched in gene sets related to immune activation and inflammatory responses. CD38 and CD200 were differentially expressed between MCL and MALD1 and confirmed by flow cytometry (median CD38, 89% vs. 14%; median CD200, 0% vs. 24%, respectively). Assessment of both proteins allowed classifying 85% (11 of 13) of MALD1 cases whereas 15% remained unclassified. SOX11 expression by qRT-PCR was significantly different between MCL and MALD1 groups but did not improve the classification. We show for the first time that MALD1, in contrast to MCL, is characterized by immune activation and driven by inflammatory cues. Assessment of CD38/CD200 by flow cytometry is useful to distinguish most cases of MALD1 from MCL in the clinical setting. MALD1 should be identified and segregated from the current MCL category to avoid overdiagnosis and unnecessary treatment.

Diffuse large B-cell lymphoma and mantle cell lymphoma of the ocular adnexal region, and lymphoma of the lacrimal gland: an investigation of clinical and histopathological features

Diffuse large B-cell lymphoma (DLBCL) and mantle cell lymphoma (MCL) constitute two distinct subtypes of non-Hodgkin lymphoma (NHL) displaying marked diversities with regard to molecular biology and clinical features. Because these differences could be related to the location of the lymphoma, it is of interest to investigate the clinical and histopathological features of DLBCL and MCL involving the ocular adnexal region (i.e. the orbit, eyelids, conjunctiva, lacrimal gland and lacrimal sac). Similarly, the lacrimal gland is the only glandular structure within the orbit. Because the lacrimal gland represents an important part of the immunological system, it is of interest to investigate lymphomas involving this location with regard to clinical and histological characteristics. The purpose of this thesis was to characterize the clinical and histopathological features of Danish patients with DLBCL of the ocular adnexal region between 1980 and 2009 and of Danish patients with ocular adnexal MCL from 1980 to 2005 (Rasmussen 2013). Furthermore, the aim was to review all specimens from patients with lacrimal gland lymphoma in Denmark between 1975 and 2009 to determine the distribution of lymphoma subtypes of the lacrimal gland and to describe the clinicopathological features of these patients. A total of 34 patients with DLBCL and 21 with MCL of the ocular adnexal region were identified (Rasmussen et al. 2009, 2013). Twenty-seven patients had lacrimal gland lymphoma (Rasmussen et al. 2011). Elderly patients predominated in all three groups, with median ages of 78, 75 and 69 years in the DLBCL, the MCL and the lacrimal gland lymphoma groups, respectively. Patients with MCL had a preponderance of men (Rasmussen et al. 2009), whereas women prevailed among patients with lacrimal gland lymphoma (Rasmussen et al. 2011). The orbit was the most common site of involvement in DLBCL and MCL. The majority of lacrimal gland lymphomas were low grade, and the distribution of subtypes was as follows: extranodal marginal zone lymphoma, 10 (37%); follicular lymphoma, 5 (19%); DLBCL, 4 (15%); MCL, 3 (11%); chronic lymphocytic leukaemia/small lymphatic lymphoma, 2 (7%); and unclassified B-cell lymphoma, 3 (11%) (Rasmussen et al. 2011). The 5-year overall survival rate for the DLBCL and MCL groups was 20% and 22%, respectively (Rasmussen et al. 2009; Rasmussen 2013). However, 83% of the patients with MCL who received rituximab-containing chemotherapy were alive after 5 years from the time of diagnosis. The 5-year overall survival rate of patients with lacrimal gland lymphoma was 70% (Rasmussen et al. 2011). In conclusion, DLBCL and MCL involving the ocular adnexal region, and lymphoma of the lacrimal gland are prevalent among elderly patients. The overall prognosis in DLBCL and MCL was poor, whereas the prognosis for patients with lacrimal gland lymphoma was relatively good. Chemotherapy containing rituximab significantly improved survival in patients with MCL. Note from the Editor: This PhD thesis is published as an electronic supplement, see: Rasmussen PK (2013): Diffuse large B-cell lymphoma and mantle cell lymphoma of the ocular adnexal region, and lymphoma of the lacrimal gland: an investigation of clinical and histopathological features. Acta Ophthalmol. 91 (thesis 5): 1–27

Elevated Soluble IL-2Ra Levels Are Associated With Inferior Outcome and Is Independent Of MIPI Score in Patients With Mantle Cell Lymphoma

Blood cytokine elevations have been associated with non-Hodgkin's lymphoma (NHL) such as follicular lymphoma (FL) and diffuse large B-cell lymphoma (DLBCL). Although other types of NHL have been studied, to our knowledge, cytokine deregulation in mantle cell lymphoma (MCL) has not been comprehensively studied. In this study, we studied cytokine levels in untreated and relapsed MCL patients using a multiplex assay to determine the role of dysregulated cytokines in these patients and study any prognostic relevance that could be drawn from their effect on relapse and survival.Samples from two data sets of MCL patients were used for this study. The first dataset was pre-treatment serum samples from newly-diagnosed patients with MCL (n=88) from the University of Iowa/Mayo Clinic SPORE Molecular Epidemiology Resource. The second dataset included pre-treatment plasma samples from relapsed MCL patients (n=20) treated on the MC0048G clinical trial studying the effect of everolimus in lymphoma patients. In addition, unrelated controls from a case-control study of lymphoma were included (n=15 serum, n=24 plasma). Thirty cytokines were assessed using a multiplex ELISA. Differences between cytokines in cases and controls were assessed using Wilcoxon rank-sum tests; associations between cytokines and event-free survival were assessed using Kaplan-Meier curves and Cox proportional hazards models. In the newly diagnosed MCL patients, IL-10 (p<0.0001), IL-1RA (p=0.001), IL-6 (p= 0.04), sIL-2Rα (p=0.005), and MIG (p=0.002) were elevated compared to normal controls. The only significantly elevated cytokine in the recurrent MCL cases compared to controls was sIL-2Rα (p= 0.0006). High levels (above median) of sIL-2Rα levels were associated with inferior event-free-survival (EFS) in both newly diagnosed (HR=2.61 (95% CI: 1.41-4.81, p= 0.0022) and relapsed MCL (HR=2.90, 95% CI: 1.08-7.80, p=0.035) patients; this remained significant after adjusting for MIPI (p=0.01 and 0.03, respectively). Moreover, in the relapsed MCL group eotaxin (p< 0.0001), IL-12 (p< 0.0001), and MCP-1 (p= 0.0002) were suppressed compared to normal controls. We hypothesized that STAT3 (pSTAT3) in MCL cells might be the possible cause behind IL-12 suppression. Indeed, 12/13 MCL samples from IL-12-suppressed relapsed MCL patients were pSTAT3+. When studying the association between cytokine levels and clinical characteristics in both untreated and relapsed MCL groups, sIL-2R was significantly associated with the MIPI score in both untreated (p<0.0001) and relapsed (p=0.007) MCL groups; and with performance status (p=0.005), lymphoma stage (p=0.004), and B symptoms (p=0.006) in the newly diagnosed MCL group. [Display omitted] [Display omitted] Our study shows that cytokines are deregulated in both untreated and relapsed MCL patients. It emphasizes on the importance of sIL-2R in its association with negative clinical parameters and worse prognosis in MCL. These findings gives the rational for using sIL-2R along with MIPI and other parameters in predicting prognosis and further to monitor response in MCL. Furthermore, we showed that IL-12 is suppressed in relapsed MCL group and that pSTAT3 expression is a possible mechanism behind this suppression. Further studies are needed to investigate the effect of drugs that suppress pSTAT3 expression in IL-12 suppressed MCLs and disease progression in relapsed MCL patients. Disclosures:No relevant conflicts of interest to declare.

Gene Expression Profiling Reveals The Lymph Node Microenvironment As a Niche For BCR Engagement, NFκB Pathway Activation, and Tumor Proliferation In Mantle Cell Lymphoma

Mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL) are both mature-looking CD5+ B-cell malignancies that invade the peripheral blood (PB), bone marrow (BM), and secondary lymphoid organs. Both are incurable with the exception of transplant. However, in many regards MCL and CLL appear to be different: MCL is aggressive and treated at diagnosis, while CLL is observed and only treated for symptomatic disease. MCL is characterized by the t(11;14)(q13;q32) resulting in cyclin D1 overexpression, anomalies in cell cycle and DNA repair, and recently by the aberrant expression of SOX11. CLL by contrast, lacks a characteristic mutation or translocation but has been recognized for decades to have a bias for certain IGHV genes, indicating that antigen-BCR interaction is a key pathogenic pathway. More recently, MCL also showed a significant IGHVbias and that many MCL cells, like CLL, express virtually identical BCRs, so called “stereotyped BCRs”, that are predicted to recognize distinct antigens (Hadzidimitriou, Blood 2011).Comparing purified CLL cells isolated concomitantly from the PB, BM, and lymph nodes (LN), we showed that LN resident cells activates SYK and upregulate BCR-target genes (Herishanu, Blood 2011). This indicates that antigenic signaling continues throughout the disease course and that the BCR is engaged primarily in the LN. Here we used gene expression profiling (Affymetrix HU133 plus) to interrogate whether MCL, similar to CLL, displays changes in tumor biology in different anatomic compartments. We collected a total of 100 PB and LN samples from previously untreated CLL and MCL patients. The samples fall into 5 groups: MCL-PB (purified CD19, n=13), MCL-LN (biopsy, n=29), CLL-PB (purified CD19, n=24), CLL-LN (biopsy, n=17), and CLL-LNT (purified CD19, n=17).An unsupervised hierarchical clustering of all samples grouped them according to tissue derivation and disease entity into MCL-LN, CLL-LN, MCL-PB, CLL-purified tumor (either PB or LNT). The previously defined MCL-LN signature (Rosenwald, Cancer Cell 2003) distinguished MCL (PB and LN) from each of the CLL groups (2-fold change, p<0.0001). Conversely a CLL specific gene signature (Jelinek, Mol Cancer Res 2003) was preferentially expressed in the 3 CLL groups than in MCL groups (3-fold change, p<0.001). There was no statistically significant difference in the expression of these signatures between PB or LN samples of the same disease entity.To study the effect of the microenvironment (ME) on tumor cells, we compared a set of three well-defined gene signatures (BCR, NFκB, and MCL-proliferation) across our samples spectrum. An individual signature score was generated for each of the five studied groups, and compared to each other using one-way ANOVA.All three signatures scored higher in LN samples (MCL or CLL) relative to PB (average 2-fold increase, p<0.01-0.0001). These scores were comparable between MCL-LN, CLL-LN, and CLL-LNT (p=ns), suggesting that first, these signatures reflect effects on resident tumor cells, and second, this effect is similar between MCL and CLL. The proliferation signature used is linked to aggressive disease in MCL (Rosenwald et al, Cancer Cell 2003), and unexpectedly scored equally across all LN groups, taking into consideration the indolent nature of CLL. This was validated using the E2F proliferation signature in CLL (Herishanu, Blood 2011). Both proliferation scores were not different between MCL-PB and CLL-PB. However, there was some variability in BCR and NFκB scores within the MCL-PB group: A subset with high BCR and NFκB scores (comparable to LN), and another with low scores (comparable to CLL-PB). The two subsets were well separated suggesting that the BCR signaling may contribute significantly to NFκB activation.Activation of MCL tumor cells in LN was confirmed by flow cytometry showing an increase in the activation markers (CD38, CD69, and CD80) and a decrease in the chemokine receptor CXCR4 in LN resident compared to circulating cells in paired samples (n=3).These data suggest that MCL, like CLL, relies on the LN ME for BCR engagement, cell activation and proliferation, and that both may be critically dependent on BCR signaling. This conclusion is underscored by the equally dramatic clinical responses achieved with the BTK inhibitor ibrutinib.Supported by the Intramural Research Program of NHLBI and NCI, NIH. We thank our patients for donating samples to make this research possible. Disclosures:No relevant conflicts of interest to declare.

Diffuse large B‐cell lymphoma and mantle cell lymphoma of the ocular adnexal region, and lymphoma of the lacrimal gland: An investigation of clinical and histopathological features

Diffuse large B-cell lymphoma (DLBCL) and mantle cell lymphoma (MCL) constitute two distinct subtypes of non-Hodgkin lymphoma (NHL) associated with considerable morbidity and mortality. Marked diversities with regard to molecular biology and clinical features are recognized in different subsets of the two lymphomas. Because these differences could be related to the location of the lymphoma, it is of interest to investigate the clinical and histopathological features of DLBCL and MCL involving the ocular adnexal region (i.e. the orbit, eyelids, conjunctiva, lacrimal gland and lacrimal sac). Similarly, the lacrimal gland is the only glandular structure within the orbit. Because the lacrimal gland represents an important part of the immunological system, it is of interest to investigate lymphomas involving this location with regard to clinical and histological characteristics. To characterize the clinical and histopathological features of Danish patients with DLBCL of the ocular adnexal region between 1980 and 2009 and of Danish ocular adnexal MCL patients from 1980 to 2005. Furthermore, the aim of this PhD was to review all specimens from patients with lymphoma of the lacrimal gland in Denmark between 1975 and 2009 to determine the distribution of lymphoma subtypes of the lacrimal gland and to describe the clinicopathological features of these patients. A total of 34 patients with DLBCL and 21 with MCL of the ocular adnexal region were identified. Twenty-seven patients had lacrimal gland lymphoma, including four DLBCLs and three MCLs from studies I and II. Elderly patients predominated in all three groups, with median ages of 78, 75 and 69 years in the DLBCL, the MCL and the lacrimal gland lymphoma groups, respectively. MCL patients had a preponderance of males, whereas females prevailed among lacrimal gland lymphoma patients. The orbit was the most common site of involvement in DLBCL and MCL. Most DLBCL patients had unilateral involvement, while MCL patients had a high frequency of bilateral involvement. Similarly, localized lymphoma was relatively frequently seen in DLBCL patients in contrast to the predominance of disseminated lymphoma in the MCL group. The majority of lacrimal gland lymphomas were low grade, and the distribution of subtypes was as follows: extranodal marginal zone lymphoma, 10 (37%); follicular lymphoma, 5 (19%); DLBCL, 4 (15%); MCL, 3 (11%); chronic lymphocytic leukaemia/small lymphatic lymphoma, 2 (7%); and unclassified B-cell lymphoma, 3 (11%). The overall survival rates at 3 and 5 years for the entire study group of DLBCL were 42% and 20%, whereas 58% and 22% of MCL patients were alive 3 and 5 years after the time of diagnosis. The 5-year overall survival rate of lacrimal gland lymphoma patients was 70%. Concordant bone marrow involvement and the International Prognostic Index score were predictive factors for the overall survival in the DLBCL group in Cox regression analysis. Rituximab-containing chemotherapy was associated with an improved survival rate in MCL patients. Diffuse large B-cell lymphoma and MCL involving the ocular adnexal region and lymphoma of the lacrimal gland are prevalent among elderly patients. The overall prognosis in DLBCL and MCL was poor, whereas the prognosis for lacrimal gland lymphoma patients was relatively good. Concordant bone marrow involvement and the International Prognostic Index score were independent predictive factors for mortality in the DLBCL group. Chemotherapy containing rituximab significantly improved survival in the MCL group.