Factor In Mantle Cell Lymphoma Research Articles

Targeting CERS6-AS1/FGFR1 axis as synthetic vulnerability to constrain stromal cells supported proliferation in Mantle cell lymphoma.

The interaction between stromal and tumor cells in tumor microenvironment is a crucial factor in Mantle cell lymphoma (MCL) progression and therapy resistance. We have identified a long non-coding RNA, CERS6-AS1, upregulated in MCL and associated with poor overall survival. CERS6-AS1 expression was elevated in primary MCL within stromal microenvironment and in a subset of MCL cells adhered to stromal layer. These stromal-adhered MCL-subsets exhibited cancer stem cell signatures than suspension counterparts. Mechanistically, we found that downregulating CERS6-AS1 in MCL reduced Fibroblast Growth Factor Receptor-1 (FGFR1), expression attributed to loss of its interaction with RNA-binding protein nucleolin. In addition, using in-silico approach, we have discovered a direct interaction between nucleolin and 5'UTR of FGFR1, thereby regulating FGFR1 transcript stability. We discovered a positive association of CERS6-AS1 with cancer stem cell signatures, and Wnt signaling. Building on these, we explored potential therapeutic strategies where combining nucleolin-targeting agent with FGFR1 inhibition significantly contributed to reversing cancer stem cell signatures and abrogated primary MCL cell growth on stromal layer. These findings provide mechanistic insights into regulatory network involving CERS6-AS1, nucleolin, and FGFR1 axis-associated crosstalk between tumor cells and stromal cell interaction and highlights therapeutic potential of targeting a non-coding RNA in MCL.

Soluble CD163 predicts outcome in both chemoimmunotherapy and targeted therapy-treated mantle cell lymphoma.

The outcome for patients with mantle cell lymphoma (MCL) has drastically improved with new treatments directed toward the tumor immune microenvironment, where macrophages play an important role. In MCL, the presence of M2 macrophages defined by CD163 expression in diagnostic biopsies has been associated with a worse prognosis. An alternative way to assess the abundance of M2 macrophages is by measuring the level of soluble CD163 in serum (sCD163). We aimed to investigate the prognostic value of sCD163 in 131 patients with MCL. We found that high sCD163 at diagnosis was associated with shorter progression-free survival (PFS) and shorter overall survival (OS) in 81 patients who were newly diagnosed and subsequently treated with chemoimmunotherapy. The same was seen in a cohort of 50 patients with relapsed MCL that were mainly treated within the phase 2 Philemon-trial with rituximab, ibrutinib, and lenalidomide. In patients who were newly diagnosed and had low levels of sCD163, 5-year survival was 97%. There was a moderate correlation between sCD163 and tissue CD163. The association with a poor prognosis was independent of MCL international prognostic index, Ki67, p53 status, and blastoid morphology, as assessed in a multivariable Cox proportional hazards model. In this study, high sCD163 was associated with both shorter PFS and shorter OS, showing that high levels of the M2 macrophage marker sCD163 is an independent negative prognostic factor in MCL, both in the chemoimmunotherapy and ibrutinib/lenalidomide era. In addition, low sCD163 levels identify patients with MCL with a very good prognosis.

MYC protein is a high-risk factor in mantle cell lymphoma and identifies cases beyond morphology, proliferation and TP53/p53 - a Nordic Lymphoma Group study.

The transcription factor MYC is a well-described oncogene with an important role in lymphomagenesis, but its significance for clinical outcome in mantle cell lymphoma (MCL) remains to be determined. We performed an investigation of the expression of MYC protein in a cohort of 251 MCL patients complemented by analyses of structural aberrations and mRNA, in a sub-cohort of patients. Fourteen percent (n=35) of patients showed high MYC protein expression with >20% positive cells (MYChigh), among whom only one translocation was identified, and 86% (n=216) of patients showed low MYC protein expression. Low copy number gains of MYC were detected in ten patients, but with no correlation to MYC protein levels. However, MYC mRNA levels correlated significantly to MYC protein levels with a R2 value of 0.76. Patients with a MYChigh tumor had both an independent inferior overall survival and an inferior progression-free survival (hazard ratio [HR]=2.03, 95% confidence interval [95% CI]: 1.2-3.4 and HR=2.2, 95% CI: 1.04-4.6, respectively) when adjusted for additional high-risk features. Patients with MYChigh tumors also tended to have additional high-risk features and to be older at diagnosis. A subgroup of 13 patients had concomitant MYChigh expression and TP53/p53 alterations and a substantially increased risk of progression (HR=16.9, 95% CI: 7.4-38.3) and death (HR=7.8, 95% CI: 4.4-14.1) with an average overall survival of only 0.9 years. In summary, we found that at diagnosis a subset of MCL patients (14%) overexpressed MYC protein, and had a poor prognosis but that MYC rearrangements were rare. Tumors with concurrent MYC overexpression and TP53/p53 alterations pinpointed MCL patients with a dismal prognosis with a median overall survival of less than 3 years. We propose that MYC needs to be assessed beyond the current high-risk factors in MCL in order to identify cases in need of alternative treatment.

Deciphering the role of MSI2 as a regulator of stem‐like properties in mantle cell lymphoma

Introduction: Mantle cell lymphoma (MCL) is one of the most aggressive mature B-cell neoplasms. MCL frequently responds to initial treatments although later the development of resistance is common, relapsing with a more aggressive disease. Several groups have isolated MCL-cancer stem cells (CSC), presenting self-renewal, clonogenic growth, tumorigenic capabilities, and resistance to standard therapies. SRY-related HMG-box gene 11 (SOX11) is expressed in progenitors and embryonic stem cells (ESC). Its overexpression has been observed in undifferentiated tumor cell populations with CSC features. In MCL, its overexpression has been associated with more aggresive behavior and worse patient’s outcome. SOX11 is regulating several oncogenic mechanisms in MCL. However, nothing is known about its possible stemness role in MCL. Methods: To search for stem-cell related genes regulated by SOX11 that may contribute to MCL biological and clinical evolution, we compared SOX11+ and SOX11- MCL primary cases differential gene expression profiling (GEP). We analyzed the prognostic value of stem-cell related genes directly regulated by SOX11 and their involvement in stemness features in MCL, using several cellular and molecular approaches. Results: We observed a significant enrichment of leukemic- and hematopoietic stem cells (HSC)-related genes in the SOX11+ compared to SOX11- MCCL subtype. The RNA-binding protein Musashi-2 (MSI2), that maintains self-renewal and prevents differentiation in ESC and HSC, emerged as one of the most significant stem cell-related gene upregulated in SOX11+ MCL primary cases. Moreover, we have demonstrated that SOX11 binds to MSI2 promoter and activates its expression. However, MSI2 intronic superenhancers might be also responsible for MSI2 upregulation in MCL. Moreover, we found that higher expression of MSI2 is significantly associated with poor overall survival, independently of other knows high-risk features in MCL. MSI2 knockdown (KD) or MSI2 function inhibition with Ro 08-2750 (Ro) changed the GEP, downregulating genes involved in CSC-related pathways whereas upregulating pro-apoptotic genes in MCL cells. MSI2KD or MSI2 Ro-inhibition led to suppress stemness phenotypic features, such as clonogenic growth, chemoresistance and cell survival. Moreover, MSI2KD MCL cells have reduced tumorigenic engraftment into mice bone marrow and spleen compared to control cell lines in vivo, suggesting that MSI2 is an important tumorigenic factor in MCL. Conclusions: Our findings suggest that MSI2 expression in MCL is upregulated by SOX11 binding to its promoter and to several active MSI2 intronic superenhancers. MSI2 upregulation might contribute to sustain stemness and chemoresistance to MCL cells, through the post-transcriptional regulation of stem cell-related genes, representing a novel target for therapeutic interventions in aggressive MCL. Keywords: Genomics, Epigenomics, and Other -Omics, Tumor Biology and Heterogeneity No conflicts of interests pertinent to the abstract.

Deep Learning–Based Nuclear Morphometry Reveals an Independent Prognostic Factor in Mantle Cell Lymphoma

Blastoid/pleomorphic morphology is associated with short survival in mantle cell lymphoma (MCL), but its prognostic value is overridden by Ki-67 in multivariate analysis. Herein, a nuclear segmentation model was developed using deep learning, and nuclei of tumor cells in 103 MCL cases were automatically delineated. Eight nuclear morphometric attributes were extracted from each nucleus. The mean, variance, skewness, and kurtosis of each attribute were calculated for each case, resulting in 32 morphometric parameters. Compared with those in classic MCL, 17 morphometric parameters were significantly different in blastoid/pleomorphic MCL. Using univariate analysis, 16 morphometric parameters (including 14 significantly different between classic and blastoid/pleomorphic MCL) emerged as significant prognostic factors. Using multivariate analysis, Biologic MCL International Prognostic Index (bMIPI) risk group (P=0.025), low skewness of nuclear irregularity (P=0.020), and high mean of nuclear irregularity (P=0.047) emerged as independent adverse prognostic factors. Additionally, a morphometric score calculated from the skewness and mean of nuclear irregularity (P=0.0038) was an independent prognostic factor in addition to bMIPI risk group (P=0.025), and a summed morphometric bMIPI score was useful for risk stratification of patients with MCL (P=0.000001). These results demonstrate, for the first time, that a nuclear morphometric score is an independent prognostic factor in MCL. It is more robust than blastoid/pleomorphic morphology and can be objectively measured.

Final Results of a CTEP Sponsored Phase I Study of Alistertib in Combination with Bortezomib and Rituximab in Relapsed and Refractory Mantle Cell and Low Grade Non-Hodgkin Lymphoma

Background: Over-expression of Aurora A has been shown in many subtypes of non-Hodgkin lymphoma (NHL) and is a negative prognostic factor in mantle cell lymphoma (MCL), where expression correlates with tumor proliferative signature (Ki67), and may contribute to cell cycle dysregulation. Aurora A regulates chromosomal stability through effects on mitosis and cell cycle regulation. Bortezomib increases both pro-apoptotic proteins and cell cycle dependent kinase inhibitors. Thus, there is a strong scientific rationale to combine Aurora A kinase and proteasome inhibition. We evaluated the combination of alistertib, an oral Aurora A kinase inhibitor, with bortezomib and rituximab in a CTEP sponsored phase 1 study of patients with relapsed /refractory (RR) low grade NHL or MCL.Methods: Patients with RR low grade NHL or MCL, who had received at least one line of immuno-chemotherapy were treated according to a 3 + 3 design with 3 dose escalation cohorts (DL1, DL2, and DL3) to identify the recommended phase 2 dose (RP2D) with an expansion cohort at the RPTD. Patients received alisertib 30mg BID (DL1 and DL2) or 40mg BID (DL3) on days 1-7, bortezomib subcutaneous 1mg/m2 (DL1) or 1.3 mg/m2 (DL2 and DL3) on days 1, 8, and 15, and rituximab 375mg/m2 on day 1 for the first 8 cycles, and subsequently q 3 months. An expansion cohort was treated at DL3. Treatment cycles were 28 days. Dose Limiting Toxicity (DLT) was defined within the first cycle of therapy.Results: A total of 24 patients at 3 sites were enrolled between 06/2013 and 02/2018, including DL1: 3, DL2: 7; DL3: 14. Median age was 64 (range 46-87). Twelve patients (50%) were male. Nineteen patients had low grade NHL, and 5 patients had MCL. Performance status was 0-1 in 22 (92%) patients, and 2 in 2 (8%) patients.Dose Escalation and Safety: The recommended phase II dose (RP2D) was DL3, with the only DLT in DL3 (prolonged grade 4 neutropenia). The most common grade 3-4 AEs at all dose levels were neutropenia (34%), thrombocytopenia (13%), anemia (8%), and fatigue (8%); grade 2 alopecia occurred in 37% (Table 1). Other grade 4 AEs included hypertension & hypotension, respiratory failure, hyperkalemia and hyperuricemia (n=1 (4%) each); one patient died of GI hemorrhage deemed unrelated to study treatment (NSAID overdose). Grade 3 neutropenia and leukopenia was seen in 4 patients and thrombocytopenia in 1. There was one grade 3 AE each of: hypertension, pulmonary hypertension, pneumonitis, fatigue, diarrhea, infusion reaction, lung infection, and febrile neutropenia.Efficacy: A total of 18 patients were evaluable for response. Five patients were not evaluable for response for the following reasons: withdrawal of consent (3, 1 each DL1, 2, 3), non-compliance (1 DL2), and death unrelated to study drug (1, DL2). For patients who were treated with at least 3 cycles of therapy, and had a restaging CT scan, the median duration of therapy was 142 days, (range 29-590). Response was seen in 7/18 evaluable patients for an overall response rate (ORR) of 38%; 4 of 18 patients (22%) had a complete response (CR), and 4 of 18 patients (22%) had a partial response (PR). An additional 8/18 patients (44%) had stable disease for a clinical benefit rate of 88%. With a median follow-up of 30.9 months, the median PFS was 6.9 months (95%CI 4.1-28.0) and median OS not reached. OS at 3 years was 64.9% (95% CI 44.1-95.3%; Fig. 1). Median duration of response was 14.1 mo (95% CI 2.63-NR).Conclusion: Alisertib in combination with bortezomib and rituximab is a well-tolerated regimen with significant and durable activity in heavily pretreated low grade NHL and MCL. The RP2D is DL3 (R + bortezomib 1.3mg/m2 and 1.8 alisertib 40mg bid). A correlation of Aurora Kinase tumor expression with clinical outcome is planned. [Display omitted] DisclosuresBarta:Janssen: Membership on an entity's Board of Directors or advisory committees; Merck, Takeda, Celgene, Seattle Genetics, Bayer: Research Funding. Diefenbach:Merck: Consultancy, Research Funding; Genentech: Consultancy; Incyte: Research Funding; Millenium/Takeda: Research Funding; Bristol-Myers Squibb: Consultancy, Research Funding; Acerta: Research Funding; Seattle Genetics: Consultancy, Research Funding; Trillium: Research Funding; Denovo: Research Funding.

SOX11, a key oncogenic factor in mantle cell lymphoma.

SOX11 has emerged as a key transcription factor in the pathogenesis of mantle cell lymphoma (MCL) whereas it is not expressed in normal B cells or virtually in any other mature B-cell neoplasm. This review will examine the role of SOX11 as a biomarker in MCL, the new information on its transcriptional targets, and the mechanisms regulating its expression in MCL. SOX11 is highly expressed in conventional MCL, including cyclin D1-negative cases, but it is not expressed in the indolent leukemic nonnodal MCL subtype. These two MCL subtypes also differ in their cell-of-origin, IGHV mutational status and genomic instability. SOX11 promotes tumor growth of MCL cells in vivo and regulates a broad transcriptional program that includes B-cell differentiation pathways and tumor-microenvironment interactions, among others. The mechanisms upregulating SOX11 in MCL are not well understood but are mediated in part by the three-dimensional reconfiguration of the DNA, bringing together a distant enhancer region and the SOX11 promoter. SOX11 is a relevant element in the pathogenesis of MCL and has been instrumental to identify two distinct clinicobiological subtypes of this tumor. Further studies should clarify the mechanisms mediating its oncogenic potential and leading to its intriguing expression in these tumors.

Prognostic relevance of the Ki-67 proliferation index in patients with mantle cell lymphoma.

BackgroundA high Ki-67 proliferation index (PI) in neoplastic cells is associated with poor survival in mantle cell lymphoma (MCL). We aimed to determine the cut-off values for the Ki-67 PI as a prognostic factor in MCL according to bone marrow findings.MethodsImmunohistochemical (IHC) staining for Ki-67 was performed on formalin-fixed paraffin-embedded biopsy tissues from 56 patients with MCL. Patients were grouped based on their Ki-67 PI values. Survival analyses were carried out and the cut-off value for the Ki-67 PI was determined.ResultsOf the 56 patients, 39 (69.6%) showed bone marrow involvement of MCL; 21 of these patients had leukemic manifestations at the time of diagnosis. The results of the Ki-67 IHC staining were as follows: ≤10% in 22 patients, 11–20% in 14 patients, 21–30% in 3 patients, 31–40% in 4 patients, 41–50% in 4 patients, and >50% in 9 patients. A cut-off value of 20% revealed significantly different survival rates with mean survival times of 69.8 months (Ki-67 PI≤20%) and 47.9 months (Ki-67 PI>20%), irrespective of bone marrow findings (P=0.034). Clinical outcomes did not differ, regardless of bone marrow findings. However, in cases with bone marrow involvement, the Ki-67 cut-off value of 30% for overall survival was required to yield statistical significance (P=0.033).ConclusionThe 20% cut-off value for the Ki-67 PI was clinically meaningful, regardless of bone marrow involvement of MCL. For patients with bone marrow involvement, the statistically significant cut-off value increased to 30%.

Identification of miR-15b as a transformation-related factor in mantle cell lymphoma.

Mantle cell lymphoma (MCL) is an aggressive B cell lymphoma with a poor prognosis. It is characterized by the t(11;14)(q13;q32) translocation, resulting in overexpression of CCND1. Morphologically, MCL is categorised into two types: classical MCL (cMCL) and aggressive MCL (aMCL), with a proportion of cMCL progressing to develop into aMCL. miRNAs are currently considered to be important regulators for cell behavior and are deregulated in many malignancies. Although several genetic alterations have been implicated in the transformation of cMCL to aMCL, the involvement of miRNAs in transformation is not known. In an effort to identify the miRNAs related to the transformation of MCL, miRNA microarray analyses were used for cMCL and aMCL cases. These analyses demonstrated significant differences in the expression of seven microRNAs based on a t-test (p-value <0.05); miR-15b was greatly upregulated in aMCL. Locked nucleic acid in situ hybridization showed increased staining of miR-15b in formalin-fixed paraffin-embedded sections of aMCL. These results correlated well with the microRNA microarray analysis. Although the molecular functions of miR-15b are largely unknown, it has been found to be associated with the cell cycle and apoptosis. However, the physiological significance of increased miR-15b in MCL is still unknown. Our present findings suggest that the upregulated expression of miR-15b is likely to play an important role in the transformation of cMCL to aMCL.

Expression of LATS mRNA in mantle cell lymphoma and its clinical significance

To investigate the expression level of large tumor suppressor (LATS)1, LATS2 mRNA and its prognostic value in mantle cell lymphoma (MCL). A total of 36 B-NHL cases (including MCL 16 cases, chronic lymphoblastic leukemia (CLL) 11 cases, splenic marginal zone cell lymphoma (SMZL) 9 cases) and 8 healthy donors were enrolled in this study from January 2008 to April 2011 in our Lymphoma Clinic Center. The mRNA level of Yap (effector of Hippo pathway) and LATS1, LATS2 mRNA were detected by using real-time quantitative PCR. Log expression values of real-time quantitative PCR data were used in this analysis. The YAP mRNA expression level in MCL, CLL and SMZL patient were significantly higher than that in health donor (lg: 1.97±0.79, 1.83±0.54, 2.12±0.42 vs 1.21±1.56, all P<0.05). The expression level of LATS1, LATS2 mRNA in MCL was significantly correlated with molecular cytogenetic aberrations, progression free survival (PFS) and overall survival (OS). The LATS1 expression level was statistically lower in the group of MCL with delection P53 than the group of MCL without delection P53 (lg: 0.75±0.27 vs 1.10±0.19, P=0.035). And the LATS1 expression level was statistically lower in the death group than the survival group (lg: 0.76±0.27 vs 1.15±0.17, P=0.026). The PFS and OS were significantly longer in the MCL patients with high level of LATS1 than that of other MCL patients ((70.4±32.7) vs (5.6±2.2) months, P=0.044; (123.8±22.0) vs (7.7±2.2) months, P=0.017). Hippo pathway is dysfunctional in B-NHL, especially in MCL.The reduced expression of LATS1, LATS2 in MCL patients is associated with progressive disease, and might be an important prognostic factor in MCL.

Tumor Cell Proliferation (Ki-67 Index) Overcomes Cytology and Growth Pattern As Prognostic Factor in Mantle-Cell Lymphoma – Results from Randomized Trials of the European MCL Network

On Behalf of the European MCL Pathology PanelIntroduction: Mantle-cell lymphoma (MCL) is an aggressive B-cell lymphoma with a median overall survival (OS) of 5 years, but clinical course varies considerably. This variability has been partly explained by clinical characteristics forming the MIPI (Hoster et al., JCO 2014), but also biological and histological features such as tumor cell proliferation (Ki-67 index), cytology, or growth pattern (Tiemann et al., BJH 2005). Immuno-chemotherapy induction represents the current standard of care, in younger patients including high-dose cytarabine and followed by autologous stem cell transplantation, whereas older patients benefit from rituximab maintenance (Dreyling et al., Ann Oncol 2013). In 2004, the European MCL Network started two large randomized trials, MCL Younger and MCL Elderly, for previously untreated MCL patients. Histopathological features of diagnostic samples were centrally assessed by the European MCL Pathology Panel. Since MCL is relatively rare, evaluations of prognostic factors were mostly based on smaller, retrospectively collected patient cohorts. We now aimed to comparatively evaluate the prognostic value of Ki-67 index, cytology, and growth pattern using the data of these trials.Methods: The Ki-67 index was counted according to published guidelines (Klapper et al., J Hematopathol 2009). MCL cytology was classified as classic, small-cell (B-CLL-like), pleomorphic (DLBCL-like), or blastic (LB-like) (Vogt and Klapper, Histopathology 2013). MCLs with pleomorphic or blastic cytology were combined to blastoid MCL. The growth pattern was classified as diffuse (≤50% nodular) or non-diffuse (>50% nodular or predominantly mantle-zone pattern). The prognostic relevance of these markers was evaluated with respect to OS, adjusting for MIPI score.Results: Of 1012 trial patients with MCL, Ki-67 index, cytology, or growth pattern were available in 50%, 61%, and 47%, respectively. Reasons for missing information were mainly insufficient material or staining. Median Ki-67 index was 20% (2%-97%). 88% had classic MCL, 2% small-cell, 7% pleomorphic and 3% blastic cytology, summing up to 10% with blastoid MCL. 63% of patients had a diffuse growth pattern. Higher Ki-67 index, blastoid MCL, and diffuse growth were each associated with higher MIPI score. Growth pattern was not clearly associated with Ki-67 index or cytology, whereas pleomorphic (median Ki-67 index 39%, range 7%-90%) or blastic MCL (median 80%, 29%-97%) displayed a substantially higher Ki-67 index compared non-blastoid MCL (median 19%, 2%-95%). In univariable analyses, the adjusted OS hazard ratios for higher Ki-67 index (10% increase), blastoid MCL, or diffuse growth were 1.16 (95% CI 1.09-1.24, p<0.0001), 1.91 (1.34-2.72, p=0.0004), and 1.16 (0.84-1.60, p=0.38), respectively. In multivariable analyses, Ki-67 index was more relevant (adjusted hazard ratio 1.12, 1.04-1.22, p=0.0032) than blastoid MCL (1.46, 0.94-2.29, p=0.095), whereas growth pattern was not prognostic. Accordingly, patients with Ki-67 index ≥30% had substantially inferior OS than patients with Ki-67 index <30% in both, blastoid and non-blastoid MCL (Figure 1).Conclusions: In a large cohort of MCL patients treated in randomized trials according to current guidelines, tumor cell proliferation (Ki-67 index) was a powerful prognostic marker, superior to cytology and growth pattern, and independent of clinical prognostic factors. The Ki-67 index further stratified patients with non-blastoid as well as patients with blastoid MCL into groups with substantially different survival. Thus, standardized assessment of the Ki-67 index (Klapper et al. J Hematopathol 2009) should be routinely performed in clinical practice to allow a more comprehensive individual risk estimation of MCL patients. Further analyses will aim at the biological basis of this prognostic effect. [Display omitted] [Display omitted] DisclosuresDreyling:Roche: Honoraria, Research Funding. Klapper:Roche: Research Funding.

Prognostic impact of proliferative index determined by quantitative image analysis and the International Prognostic Index in patients with mantle cell lymphoma

BackgroundThe proliferative index (PI) is a powerful prognostic factor in mantle cell lymphoma (MCL); however, its utility is hampered by interobserver variability. The mantle cell international prognostic index (MIPI) has been reported to have prognostic importance. In this study, we determined the prognostic value of the PI as determined by quantitative image analysis in MCL. Patients and methodsEighty-eight patients with adequate tissue were included in this analysis. Patients were treated with one of two treatment programs: sequential therapy with high-dose therapy consolidation or radioimmunotherapy followed by combination chemotherapy with cyclophosphamide, doxorubicin, vincristine and prednisone. Patients were divided into four groups based on PI (<10%, 10%–29.9%, 30%–49.9%, and >50%), and outcomes were analyzed. ResultsThirty percent was identified as the optimal cut-off for PI. By univariate analysis, intensive treatment and a low PI were associated with a superior progression-free survival (PFS); only PI was associated with overall survival. By multivariate analysis, both intensive treatment and PI correlated with PFS. The MIPI had no prognostic impact. ConclusionsPI is the most important prognostic factor in MCL. The cut-off of 30% is clinically meaningful and can be used to tailor the intensity of therapy in future clinical trials.

Nuclear Expression of the Sox 11 Transcription Factor in Mantle Cell Lymphoma, and Cytoplasmic Expression in Follicular Lymphoma and Multiple Myeloma: Pathogenetic Implications.

Abstract 2925Poster Board II-901Mantle cell lymphoma (MCL) is characterized by nuclear cyclin D1 expression resulting from the t(11;14)(q13;q32). As cyclin D1 overexpression alone is insufficient for B-cell transformation, we have investigated other potentially contributing mutations. Sox11 is a member of a large family of transcription factors containing a DNA-binding high-mobility group (HMG) domain and shares homology with Sox4, which is involved in lymphopoiesis. Recent reports have identified Sox11 expression in the majority of MCL, suggesting it may contribute to pathogenesis (Ek et al, Blood 2008;111:800; Wang et al, Br J Haematol 2008;143:248). Methods:Patients with MCL diagnosed at the Univ. of Virginia from 1997-2008 and for whom nodal, marrow or spleen tissue blocks were available were identified from the Pathology database. Follicular lymphoma (FL), marginal zone lymphoma (MZL), small lymphocytic lymphoma (SLL), hairy cell leukemia (HCL) and multiple myeloma (MM) were also analyzed. Formalin-fixed, paraffin-embedded samples were stained by immunohistochemistry for cyclinD1, Sox11 (rabbit a/Sox11, 1:100, Sigma) and Ki-67. Nuclear and/or cytoplasmic expression was determined by two of us (YT, JC). Results:All MCL samples revealed nuclear cyclin D1 expression. Nuclear but not cytoplasmic Sox11 expression was identified in all 24 non-blastoid and in 5/6 blastoid samples; 1 blastoid sample showed only cytoplasmic Sox11 staining (Table). Four of 5 HCL expressed cyclin D1 but were negative for Sox11; 4/5 MM also were cyclin D1-positive, with 3 positive for cytoplasmic Sox11 including 2 of the cyclin D1-positive cases. All 5 FL showed cytoplasmic Sox11 positivity, whereas all MZL and SLL were negative for cytoplasmic or nuclear staining.TypePatientsSamplesNuclear Cyclin D1+Nuclear Sox11+Cytoplasmic Sox11+MCL, non –blastoid222424240MCL, blastoid56651FL55N.D.105MZL66000SLL66000HCL255400MM3554031N.D. = Not Done23 spleen, 2 bone marrow samples3Bone marrow samples Conclusions:Nuclear Sox11 expression was uniformly identified in MCL, with the exception of cytoplasmic expression in a single blastoid case. No nuclear Sox11 was present in HCL or MM despite the expression of cyclin D1, although 3 MM showed cytoplasmic Sox11 staining. These findings suggest that nuclear Sox11 overexpression is not a direct result of dysregulated cyclin D1 signaling but instead occurs by alternative mechanisms. The significance of cytoplasmic staining remains uncertain. A potential pathogenetic role for Sox11 and possibly other Sox family transcription factors warrants further investigation in MCL and other lymphoproliferative neoplasms for diagnostic use and therapeutic targeting. Disclosures:No relevant conflicts of interest to declare.

Number of Lymphoma-Associated-Macrophages (LAM) Is An Independent Predictor of Survival in Patients with Mantle Cell Lymphoma (MCL).

Abstract 3944Poster Board III-880 BackgroundMCL is a clinically aggressive lymphoma characterized by frequent relapses and short survival. Clinical behavior and response to therapy vary considerably and are often unpredictable based on clinical factors. Prior gene and protein expression studies have demonstrated proliferation and TP53 expression as biological mechanisms that strongly influence survival (Rosenwald Cancer Cell 2003 & Katzenberger Blood 2006 and Louie Blood 1995). Recent studies have highlighted the role of the microenvironment in follicular lymphoma (FL) including numbers of lymphoma-associated-macrophages (LAM), as important prognostic factors (Dave NEJM 2004 & Farinha Blood 2005). Within most tumors, macrophages display an M2 phenotype that promotes tumor growth and angiogenesis. In MCL, the clinical impact of LAM is unknown. Our aim was to assess the prognostic impact of LAM in MCL patients from a single institution experience using tissue microarrays (TMA), immunohistochemistry and clinical correlates. MethodsA TMA block was built with duplicate 0.6mm cores of paraffin-embedded formalin-fixed diagnostic biopsies from 185 patients treated at the BC Cancer Agency (1983-2004). A subset of cases was also analyzed for the presence of the t(11;14) by FISH. Primary and subsequent therapy included multiple regimens: single or multi-agent chemotherapy with or without rituximab, stem cell transplant, radiation and therapeutic splenectomy. Standard immunohistochemistry was performed with CD20, CCND1, CD68, CD163 (an M2 marker), CD34, TP53 and Ki67. TP53 and Ki67 IHC were scored using image analysis (VIAS). Univariate and multivariate correlates of overall survival (OS) were determined using SPSS software. ResultsThe median follow-up of living patients was 5.75 years. The IPI predicted OS (p=0.0013). Numbers of cases with interpretable staining varied for the different biomarkers (range 157 to 180). All 185 cases were either CCND1 positive and/or t(11;14) FISH positive. Both proliferation (quartiles of Ki67+ cells) and TP53 (dichotomized based on cut-off >40%) were significant predictors of OS independent of IPI (OS, p=0.0002). Cases showing more than 5% infiltrating CD68+ cells (18/185) had a 4-y OS of 5% vs. 35% (p=0.0081). These CD68+ cases significantly correlated with 12 cases showing more than 5% infiltrating CD163+ cells (x2=0.001), which also significantly influenced OS (p=0.008). 17 cases had increased microvessel density (MVD), defined by CD34+ cells, which correlated with inferior OS (p=0.02). Interestingly, cases with increased MVD often had >5% CD163+ cells (x2=0.05), but not increased CD68+ cells. Multivariate analysis including IPI, TP53, Ki67, CD68, CD163 and CD34, showed IPI (RR=1.8, 95%CI=1.0-3.1, p=0.047), TP53 (RR=2.3, 95%CI=1.3-4.3, p=0.006), Ki67 (RR=2.2, 95%CI=1.2-4.2, p=0.012) and CD68 (RR=2.1, 95%CI=1.1-3.8, p=0.022) to be independent predictors of OS. Finally, in the 27 patients who received rituximab at some time during their disease course, increased CD68+ cells did not significantly affect OS (p=0.1), suggesting that, similar to FL, the prognostic effect of LAM may be abolished by rituximab. ConclusionsThe number of LAMs is an important prognostic factor in MCL, independent of clinical parameters, proliferation and TP53 expression. Expression of CD163, described as an M2 polarized macrophages, also predicts survival and correlates with increased angiogenesis. These preliminary data suggest that the use of rituximab may abrogate the prognostic impact of LAM in MCL. In summary, similar to other lymphoma subtypes, the microenvironment is important in MCL biology and prognosis and may be an important new target for therapy in this aggressive disease. [Display omitted] Disclosures:Connors:Roche Canada (F Hoffmann-La Roche): Research Funding. Gascoyne:Roche Canada, Genentech, Lilly, Millennium : Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.

Five-Gene Model to Predict Survival in Mantle-Cell Lymphoma Using Frozen or Formalin-Fixed, Paraffin-Embedded Tissue

Despite the common underlying translocation t(11;14) involving cyclin D1 that is present in nearly all cases of mantle-cell lymphoma (MCL), the clinical course of the disease is highly variable. The aim of the present study was to develop a quantitative gene expression-based model to predict survival in newly diagnosed patients with MCL that involves a minimum number of genes and is applicable to fresh-frozen and formalin-fixed, paraffin-embedded (FFPE) tumor samples. The expression of 33 genes with potential prognostic and pathogenetic impact in MCL was analyzed using quantitative reverse-transcription polymerase chain reactions (qRT-PCR) in a low-density array format in frozen tumor samples from 73 patients with MCL. Multivariate Cox methods and stepwise algorithms were applied to build gene expression-based survival predictors. An optimized five-gene model was subsequently applied to FFPE tumor samples from 13 patients with MCL from the initial series and to 42 independent MCL samples. The optimized survival predictor was composed of the five genes RAN, MYC, TNFRSF10B, POLE2, and SLC29A2 and was validated for application in FFPE tissue samples. It allowed the survival prediction of patients with MCL with widely disparate clinical outcome and was superior to the immunohistochemical marker Ki-67, an established prognostic factor in MCL. We here present a validated qRT-PCR-based test for survival prediction in patients with MCL that is applicable to fresh frozen as well as to FFPE tissue specimens. This test may prove useful to guide individualized treatment approaches for patients with MCL.

Repp86: a new prognostic marker in mantle cell lymphoma

Proliferation indices are important prognostic factors for the clinical outcome of patients with mantle cell lymphoma (MCL). We investigated whether the expression of repp86 (restrictedly expressed proliferation-associated protein 86 kDa), a new proliferation specific marker expressed in the cell cycle phases G(2), S and M, but not in G(1), correlates with the clinical course in patients with MCL. Biopsy specimens from 94 untreated patients enrolled in two multicenter trials were investigated immunohistochemically with monoclonal antibodies against CD20, CD5, CD3, CD23, cyclin D1, and repp86 (Ki-S2). Patients with 0-1% repp86 expression had a median overall survival time of 71.0 months, compared with 38.2 months for patients with 1-5% positive cells and 25.4 months for patients with 5-10% positive tumor cells. Patients with repp86 expression of more than 10% showed the shortest survival (median: 15.0 months). Kaplan-Meier analysis revealed a significant difference in the overall survival time between patients with very high (>10%) and very low (0-1%) repp86 expression (P < 0.0001) in the tumor cells. The multivariate analysis revealed repp86 expression to be superior to other clinical characteristics as a prognostic factor (P = 0.0016). Based on these findings, repp86 expression is a new important prognostic factor in MCL.

Topoisomerase IIalpha as a prognostic factor in mantle cell lymphoma.

Topoisomerase IIalpha as a prognostic factor in mantle cell lymphoma.