A Novel Anti-IgM/HLA-DR Bispecific Antibody for Treatment of Refractory B Cell Malignancies

Abstract Introduction: IgM composes a part of the B cell receptor, and its expression is restricted to B cell lineage cells. There have been several reports that refractory B cell malignant cells frequently express IgM. For example, activated B cell-like (ABC) diffuse large cell lymphoma (DLBCL) is a subtype with a poor prognosis following R-CHOP treatment (Lenz et al., N Engl J Med, 2008), and the tumor cells generally express IgM-type immunoglobulin (Ruminy et al., Leukemia, 2011). In addition, IgM-expressing chronic lymphocytic leukemia (CLL) was also found to have a poor prognosis (D'Avola et al., Blood, 2016). Therefore, IgM is a promising therapeutic target for refractory B cell malignancies. However, anti-IgM antibodies may not function fully in the body due to the presence of soluble IgM. To improve the selectivity to the IgM expressing cells, we produced a bispecific antibody bound to IgM and HLA-DR, which are both expressed by B cells, and it was able to bind IgM+HLA-DR+ cells in the presence of soluble IgM. Thus, the aim of the present study was to examine the in vitro anti-tumor activity of the anti-IgM/HLA-DR bispecific antibody against B cell lymphoma, and to clarify its efficacy and tolerability in non-human primates (NHP). Methods: BTA106 is the bispecific antibody that can bind to IgM and HLA-DR, and BTA111 is glycoengineered BTA106. To investigate their direct inhibitory effects and Fc-mediated effects against B cell lymphoma cell lines, a growth inhibition assay, antibody-dependent cell cytotoxicity (ADCC) assay and complement dependent cytotoxicity (CDC) assay were performed. In the growth inhibition assay and CDC assay, cell proliferation was measured by WST-8. In the ADCC assay, human peripheral blood mononuclear cells were used as effector cells. Dead cells were stained with propidium iodide and measured by flow cytometry. JeKo-1 (Mantle cell lymphoma, MCL), Ramos (Burkitt's lymphoma, BL), RL (DLBCL), B104 (B cell lymphoma) and two rituximab-resistant cell lines, RL-4RH (Czuczman et al., Clin Cancer Res., 2008) and RRBL1 (generously provided by Dr. Tomita, Tomita et al., Int J Hematol., 2007), were used. To clarify the efficacy and tolerability of BTA106 in NHP, a dose escalation study and single-dose administration study were performed using cynomolgus monkeys. Results: BTA106 and BTA111 demonstrated direct growth inhibition activity, and CDC and ADCC activity, although their intensities differed among cell lines. In the case of rituximab-resistant cell lines, RRBL1 and RL-4RH, BTA106 and BTA111 exhibited weak or no growth inhibitory effects, but their CDC and ADCC activity was superior to that of rituximab. Development of resistance to rituximab is also a poor prognostic factor and remains problematic. Thus, BTA106 and BTA111 are expected as therapeutic antibodies to overcome rituximab resistance. A dose escalation study and single dose study were performed in cynomolgus monkeys. BTA106 greatly reduced peripheral B cells. Moreover, after the administration of BTA106, lymph follicles disappeared in axillary lymph nodes. In these studies, decreased locomotor activity and vomiting were observed; however, these symptoms resolved 2 hrs after injection. Other BTA106-related clinical toxicities or changes in body weight were not observed. Conclusions: In the in vitro studies, BTA106 and BTA111 demonstrated significant anti-tumor effects on rituximab-sensitive and -resistant B cell lymphoma cell lines. The efficacy and toxicity studies in cynomolgus monkeys revealed that BTA106 depletes peripheral and lymph node B cells, and it was well tolerated. BTA111 was considered to be significantly more effective than BTA106 due to glycoengineering, which induced high ADCC activity. These data suggest that BTA111 is an effective and potent therapeutic antibody for B cell malignancies, especially rituximab-resistant B cell lymphoma. Currently, a repeated administration study of BTA111 with cynomolgus monkeys to evaluate its efficacy and toxicity is ongoing. Disclosures Ohashi: Zenyaku Kogyo Co., Ltd.: Employment. Miyashita:Zenyaku Kogyo Co., Ltd.: Employment. Nagata:Zenyaku Kogyo Co., Ltd.: Employment. Tatebe:Zenyaku Kogyo Co., Ltd.: Employment. Otsuka:Zenyaku Kogyo Co., Ltd.: Employment. Suzuki:Zenyaku Kogyo Co., Ltd.: Employment. Sasaki:Zenyaku Kogyo Co., Ltd.: Employment. Enami:Zenyaku Kogyo Co., Ltd.: Employment. Tsukada:Zenyaku Kogyo Co., Ltd.: Employment. Fukushima:Zenyaku Kogyo Co., Ltd.: Employment.

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RAD001 (everolimus) down-regulates cyclin D3 and c-Myc and is particularly effective in the treatment of aggressive B-cell lymphomas

Journal of Clinical Oncology ◽

10.1200/jco.2006.24.18_suppl.17573 ◽

2006 ◽

Vol 24 (18_suppl) ◽

pp. 17573-17573

Author(s):

S. H. Kuo ◽

C. H. Hsu ◽

P. Y. Yeh ◽

H. C. Hsu ◽

M. Gao ◽

...

Keyword(s):

Growth Inhibition ◽

B Cell ◽

Cell Lines ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Lymphoma Cell ◽

Cyclin D3 ◽

Akt Signaling Pathway ◽

P70s6 Kinase ◽

Aggressive B Cell Lymphoma

17573 Background: Aggressive B-cell lymphoma is recently found to be associated with constitutive activation of the PI3K/Akt pathway, and thereby is relatively resistant to apoptosis. Furthermore, activation of the PI3K/Akt signaling pathway can result in the upregulation of cyclin D3 and c-Myc through inhibition of the cap-independent RNA translation. Since mTOR is closely associated with the regulation of the translation process, and is a downstream mediator of the PI3K/Akt signalling pathway, this study aimed to examine if RAD001, an inhibitor of mTOR, can be effective in the treatment of aggressive B-cell lymphoma. Methods: Pfeiffer (diffuse large cell lymphoma), Ramos (EBV (−) Burkitt’s lymphoma), Raji (EBV (+) Burkitt’s lymphoma), and MC116 (EBV (−) undifferentiated lymphoma) cell lines were used in this study. Expression of pAkt, p70S6 kinase, pp70 S6 kinase, 4E-BP1, p4E-BP1, cyclin D3, and c-Myc was measured by immunoblotting. Results: Akt was constitutively activated in all four lymphoma cell lines. Downstream effectors of PI3/Akt signaling pathway, including p70S6 kinase and 4E-BP1, were also phosphorylated in these lymphoma cell lines. RAD001 downregulated p70S6 kinase and 4E-BP1, and the IC50s of growth suppression ranged from 5 to 50 nM, without significant difference between EBV (+) and EBV (−) cell lines. The IC50s of these lymphoma cell lines appeared to be much lower than those obtained from the carcinoma cell lines, suggesting that lymphomas may be particularly sensitive to growth inhibition by RAD001. RAD001 blocked cell cycle progression in G0/G1 phase in all four lymphoma cell lines while there was no significant increase in sub-G1 phases, suggesting that apoptosis was not the major mechanism of RAD001-induced growth inhibition. In addition, in parallel with the RAD001-induced growth inhibition, a dose-dependent down-regulation of the expression of cyclin D3 and c-Myc was demonstrated. Conclusions: The mechanism of action is at least partly due to downregulation of cyclin D3 and c-Myc, and subsequent G0/G1 block. Since overexpression of cyclin D3 and c-Myc is also closely associated with chemoresistance of aggressive B-cell lymphoma, RAD001 may be a useful adjunct in the treatment of this group of tumors. No significant financial relationships to disclose.

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B Cell Lymphomas Are Resistant towards Bone Morphogenetic Protein (BMP) Induced Growth Inhibition.

Blood ◽

10.1182/blood.v108.11.2381.2381 ◽

2006 ◽

Vol 108 (11) ◽

pp. 2381-2381

Author(s):

Kanutte Huse ◽

Marianne B. Eide ◽

Christian Kersten ◽

Erlend B. Smeland ◽

June H. Myklebust

Keyword(s):

Growth Inhibition ◽

B Cell ◽

Cell Lines ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Lymphoma Cell ◽

Lymphoma Cells ◽

Bmp Receptors ◽

Induced Apoptosis

Abstract Bone morphogenetic proteins (BMPs) belong to the TGF-β superfamily, and mediate their effects mainly through the Smad signalling pathway. Whereas TGF-β is well established as one of the most potent negative regulators in hematopoietic cells, the role of BMPs remains more elusive. We have previously shown that BMP-6 inhibits the growth of naïve and memory human B cells. As high BMP-6 mRNA expression is associated with poor outcome in diffuse large B cell lymphoma (DLBCL; Rosenwald et al, N Engl J Med 2002), we hypothesized that resistance towards BMP-induced growth inhibition is a possible mechanism for lymphomagenesis. In the current study, 7 B cell lymphoma cell lines (representing Burkitt lymphoma (BL) and DLBCL) and tumour material from lymphoma patients were investigated to unravel the role of BMPs in lymphomas. We analyzed the expression of BMP receptors by FACS analysis, and found variable expression of the BMP receptor type I (Alk2, Alk3 and Alk6) and type II (BMP RII, Activin RIIA and RIIB) among the cell lines and in primary lymphoma cells, suggesting variable binding of BMPs. We next investigated the effect of BMP-2, BMP-4, BMP-6 and BMP-7 on proliferation and survival of B lymphoma cell lines, and found 2 of 7 cell lines to be resistant towards BMP-2 and BMP-4 induced growth inhibition. In contrast, 4 of 7 and 7 of 7 cell lines were resistant to BMP-6 and BMP-7 induced growth inhibition, respectively. In Sudhl6 cells that were highly sensitive to BMP-2 and BMP-6 induced apoptosis and inhibition of proliferation, we demonstrated that the cytokines IL-10, CD40 Ligand and BLyS were able to counteract the negative effects induced by BMPs, while IL-2 and IL-4 were not. On the contrary, both BMP-2 and BMP-6 greatly increased anti-IgM activation induced apoptosis. In resistant lymphoma cells, the BMPs were not able to induce detectable levels or induced low levels of phosphorylated SMAD1/5/8 compared to sensitive cell lines. Low or no increase in phosphorylation of SMAD1/5/8 induced by BMPs could only partly be explained by low/ undetectable expression of BMP receptors. Hence, upregulation of inhibitory Smads (Smad6, Smad7) or mutations in receptors or Smads represent other possible mechanisms for resistance to BMPs in lymphomas, and this is currently under investigation. We also investigated if the lymphoma cells produced BMPs themselves and found that 5 of 7 cell lines and 3 of 5 primary lymphomas produced significant amounts of BMP-7. Some lymphoma cells also had detectable levels of BMP-4 and BMP-6. Our findings that lymphoma cells are resistant towards BMP-7 and to some degree BMP-6 induced growth inhibition, whereas they produce these cytokines, suggest that resistance towards BMP induced signalling in B cell lymphomas can contribute to increased tumour growth.

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Forodesine (Immucillin H, BCX-1777) Shows Activity on Mantle Cell Lymphoma Primary Cells

Blood ◽

10.1182/blood.v112.11.4997.4997 ◽

2008 ◽

Vol 112 (11) ◽

pp. 4997-4997

Author(s):

Andrea Rinaldi ◽

Emilia Ceresa ◽

Davide Rossi ◽

Gianluca Gaidano ◽

Shanta Bantia ◽

...

Keyword(s):

B Cell ◽

Mantle Cell Lymphoma ◽

Cell Lines ◽

Cell Lymphoma ◽

Annexin V ◽

B Cell Lymphoma ◽

B Cell Malignancies ◽

New Therapeutic Approaches ◽

Mantle Cell

Abstract Mantle cell lymphoma (MCL) represents a subtype of B-cell lymphoma associated with a very unfavourable clinical outcome. Currently no therapy can be considered as standard, and new therapeutic approaches are needed. Forodesine is a potent inhibitor of purine nucleoside phosphorylase (PNP), whose major role is to catalyze the cleavage of inosine, deoxyinosine guanosine, and deoxyguanosine (dGuo) to their corresponding base and sugar 1-phosphate by phosphorolysis. In the presence of deoxycytidine kinase, PNP inhibition leads to an increase in the concentration of dGuo triphosphate (dGTP), followed by inhibition of DNA synthesis and cell death by apoptosis. When combined with dGuo, forodesine has been shown to have in vitro cytotoxic activity on T-cell (T-ALL, T-PLL) and on B-cell malignancies (CLL, B-ALL), and Phase I/II trials are on going in CLL and CTCL patients. Here, we report the first data on in vitro activity of forodesine in MCL. Primary MCL cells, derived from six patients, were exposed to forodesine (0, 2, 20 μM) in combination with dGuo (0, 10, 20 μM), for 48 hrs. Cells were cultured in X-VIVO 10 medium (Cambrex) with 10% FBS. Cell viability was assessed by flow cytometry with the Annexin V - propidium iodide assay. Four patient samples (67%) showed an increase in the number of Annexin V positive cells ranging from 1.9 to 5.3 times compared to untreated cells. The effect was larger for 20 μM forodesine compared with 2 μM. There was no effect of dGuo alone and only a minimal effect of increasing dGuo concentration from 10 μM to 20 μM. Cell lines did not appear to be ideal models to evaluate the efficacy of forodesine in vitro. Three established MCL cell lines (Granta-519, Rec, JeKo1) were treated with escalating doses of forodesine, but the results were not reproducible, while the same cells showed expected IC50 values between 25–30 μM when exposed to bendamustine for 72 hrs. In conclusion, the in vitro data reported here with 4/6 MCL patients primary samples sensitive to forodesine and the results from various groups on other T- and B-cell malignancies suggest that clinical trials of forodesine in MCL may be warranted.

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Genetic Alterations of the MHC Class II Transactivator CIITA Are Frequent in Primary Mediastinal Large B-Cell Lymphoma and Associated with Diminished MHC Class II Expression

Blood ◽

10.1182/blood.v124.21.3040.3040 ◽

2014 ◽

Vol 124 (21) ◽

pp. 3040-3040

Author(s):

Anja Mottok ◽

Bruce W. Woolcock ◽

Fong Chun Chan ◽

Adele Telenius ◽

Elizabeth A. Chavez ◽

...

Keyword(s):

Protein Expression ◽

B Cell ◽

Cell Lines ◽

Mhc Class Ii ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Class Ii ◽

Coding Sequence ◽

Hla Dr ◽

Intron 1

Abstract Introduction: Constitutive MHC class II expression is a hallmark of antigen-presenting cells, including B cells, and is indispensable for the initiation of antigen specific immune responses. It has been shown that certain B cell lymphoma entities are able to evade immune recognition by downregulation of MHC molecules on the tumor cell surface. We have previously identified recurrent chromosomal rearrangements of CIITA, the master regulator of MHC class II transcription, as one possible mechanism to reduce MHC class II expression in primary mediastinal large B-cell lymphoma (PMBCL) and classical Hodgkin lymphoma (cHL) (Steidl et al., Nature 2011). Furthermore, we have recently described a 1.6kb breakpoint cluster region within intron 1 of CIITA and have shown in a small sample set of PMBCL cases that deletions, insertions and single nucleotide variants (SNV) are commonly found within this genomic region (Steidl, ASH abstract # 437, 2011). Therefore, we aimed to explore the frequency of these alterations and the correlation with CIITA and MHC class II protein expression in a larger cohort of PMBCL cases and to further characterize their functional significance. Methods: We have comprehensively analyzed 45 diagnostic PMBCL samples for the presence of coding sequence mutations as well as alterations within the promoter III region and the first 3kb of intron 1 using deep amplicon sequencing (Illumina TruSeq) and/or Sanger sequencing. In addition, we characterized the PMBCL-derived cell lines U2940 and Med-B1 by whole transcriptome paired-end sequencing (RNA-seq). To elucidate the functional consequences of the coding sequence mutations identified in these two cell lines we performed retroviral transductions of wild type CIITA and CIITA mutants in a CIITA and HLA-DR expression-negative cell line (DEV, nodular lymphocyte predominant Hodgkin lymphoma-derived). We subsequently analyzed CIITA mRNA expression using qRT-PCR and HLA-DR surface expression using flow cytometry. Furthermore, we applied immunohistochemistry (IHC) to determine expression levels of CIITA and HLA-DR in a large cohort of PMBCL samples represented on two tissue microarrays (TMA, n=149). The TMAs were also used for fluorescence in-situ hybridization (FISH) to evaluate the presence of copy number alterations or translocations of the CIITA locus. Results: FISH was interpretable in 115 samples with a CIITA break-apart (CIITA-ba) frequency of 33.9% (39/115). Correlative analyses revealed that decreased CIITA protein expression by IHC was significantly correlated with the presence of CIITA-ba (P=0.019), whereas HLA-DR expression was not correlated with CIITA-ba status alone (P=0.219). However, we could demonstrate a positive correlation between protein expression of CIITA and HLA-DR (Pearson r=0.45, P<0.0001). Within the subset of 45 PMBCL cases that were analyzed for the presence of genomic alterations, 39% were CIITA-ba positive (16/41), and in 31.8% (14/44) we observed coding sequence mutations and/or alterations affecting the promoter III region. 45.5% (20/44) of the cases presented indels and/or SNVs in intron 1. Using RNA-seq, we have detected two missense mutations in the Med-B1 cell line affecting both alleles in functionally relevant protein domains. Furthermore, we identified a novel NUBP1-CIITA fusion transcript in U2940 also harboring an SNV on the other allele resulting in the transcription of an elongated protein due to the loss of the original stop codon. Ectopic expression of these CIITA mutants in DEV, which has been shown to have undetectable levels of CIITA and HLA-DR due to a biallelic CIITA inactivation, revealed that these individual SNVs showed a diminished capability to restore HLA-DR surface expression in comparison to wild type CIITA as measured by flow cytometry. Conclusions: Here we show that the presence of CIITA rearrangements is significantly associated with low CIITA protein levels, and we could demonstrate that protein expression of CIITA and HLA-DR are positively correlated in PMBCL. Furthermore, CIITA is frequently targeted by coding sequence mutations and intronic deletions in PMBCL cell lines and clinical samples. Functional studies demonstrate that genomic alterations in CIITA contribute to downregulation of MHC class II expression in malignant lymphomas and therefore represent a potent mechanism of acquired immune privilege and escape from immune surveillance. Disclosures No relevant conflicts of interest to declare.

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Motexafin Gadolinium Enhances Rituximab-Induced Cytotoxicity Against B-Cell Lymphoma Cell Lines.

Blood ◽

10.1182/blood.v104.11.2296.2296 ◽

2004 ◽

Vol 104 (11) ◽

pp. 2296-2296 ◽

Cited By ~ 1

Author(s):

Jun Chen ◽

Jason Ramos ◽

Mint Sirisawad ◽

Richard A. Miller ◽

Louie Naumovski

Keyword(s):

Growth Inhibition ◽

B Cell ◽

Cell Line ◽

Cell Lines ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Motexafin Gadolinium ◽

Redox Active ◽

Apoptotic Effect

Abstract Motexafin gadolinium (MGd, Xcytrin®) is a tumor selective redox active drug that is cytotoxic to many hematolymphoid cell lines and chronic lymphocytic leukemia cells. Previous studies have shown that MGd oxidizes various cellular reducing metabolites and produces reactive oxygen species (ROS), which can induce apoptosis. Rituximab, an anti-CD20 antibody, is used widely in the treatment of B-cell malignancies; however, its precise mechanism of action is uncertain. Some studies have demonstrated that rituximab induces the generation of ROS in sensitive cells. We evaluated the effects of combining rituximab and MGd in the in vitro treatment of HF-1, a cell line derived from a patient with follicular lymphoma. Increases in both apoptosis and cell growth inhibition were seen with MGd plus rituximab compared to each agent used separately. Apoptosis was demonstrated by both annexin-V positivity and caspase-3 activity. Loss of mitochondrial membrane potential and PARP cleavage were also greater when the two drugs were used together than when used separately. Data analysis with CalcuSyn software showed that, with the combination, the Dose Reduction Index (DRI) was more than 1.5 for MGd and 2 to 6 for rituximab regarding both apoptotic effect and growth inhibition. Consistent with MGd as a redox active agent, peroxiredoxin oxidation was higher when MGd and rituximab were used together. Further experiments revealed no increase in uptake of MGd by rituximab treatment, nor a change in the amount of CD20 antigen on the surface of MGd treated cells. We investigated another B-cell lymphoma cell line, DHL-4 for similar cooperative activity of MGd and rituximab. While MGd alone has no substantial apoptotic effect on DHL-4 cells, it enhanced the activity of rituximab in inducing apoptosis. These in vitro findings support the combined use of MGd and rituximab in the treatment of B cell lymphoma.

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Lenalidomide Enhances the Biological Activity of Galiximab Against Non-Hogkin’s Lymphoma In Vitro and In Vivo.

Blood ◽

10.1182/blood.v110.11.2353.2353 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2353-2353

Author(s):

Nishitha Reddy ◽

Francisco J. Hernandez-Ilizaliturri ◽

Joy Knight ◽

Czuczman S. Myron ◽

Keyword(s):

Growth Inhibition ◽

B Cell ◽

Cell Lines ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Raji Cells ◽

Isotype Control ◽

Tail Vein Injection

Abstract Lenalidomide, a thalidomide analog, has dual anti-tumor activities against B-cell lymphomas and other hematological malignancies by inducing direct apoptosis in malignant cells and modulating the tumor microenviroment (angiogenesis inhibition) or the host immune response. We previously demonstrated that lenalidomide enhances rituximab-mediated antibody dependent cellular cytotoxicity (ADCC). In our current work we further studied the effects of combining lenalidomide with galiximab (IDEC114), a primatized anti-CD80 monoclonal antibody, which is undergoing clinical testing in B-cell lymphoma. To this end a panel of rituximab-sensitive or rituximab-resistant B-cell lymphoma cell lines was exposed in vitro to lenalidomide (10μg/ml) or DMSO (0.01%) with our without galiximab (10μg/ml) or isotype control antibody (10μg/ml) and incubated at 37°C, 5%CO2 over five days. Changes in DNA synthesis were determined by measuring [3H]-thymidine uptake at 24 and 48 hrs. To asses changes in galiximab-mediated ADCC, peripheral blood mononuclear cells (PBMCs) isolated from healthy volunteers were incubated with lenalidomide or control for 72 hrs. Subsequently, NHL cells were labeled with 51Cr and then exposed to galiximab or isotype control (10mg/ml) and PBMCs (Effector:Target ratio, 40:1). For in vivo studies, 6–8 week old SCID mice were inoculated with 1×106 Raji cells via tail vein injection and after a period of 72 hours, in order to allow for tumor engraftment, animals were divided into four cohorts (control, lenalidomide alone, galiximab alone, and lenalidomide + galiximab). Lenalidomide was administered intraperitoneally (i.p) at 0.5mg/kg/dose on days +3, +4, +8, +9, +13, +14, +18 and +19. Galiximab was administered via tail vein injection at 10mg/kg/dose on days +5, +10, +15 and +20. Difference in survival between treatment groups was performed by Kaplan-Meier analysis. Results: In vitro exposure of various NHL cells to lenalidomide for five consecutive days enhanced the anti-proliferative effects of galiximab against Raji cells (40% of growth inhibition) when compared to Galiximab (15% growth inhibition) or Lenalidomide (14% growth inhibition) at 24 hrs. Similar results were observed in other cell lines. In addition, an improvement in galiximab-associated ADCC was observed in lenalidomide-exposed NHL cells. In vivo treatment of SCID mice with lenalidomide in combination with galiximab led to prolongation of the median survival (39 days, 35–42 95% C.I.) compared to galiximab (28 days, 26–29 95% C.I.) or Lenalidomide (23 days, 22–25 95% C.I.) alone ((p<0.01). Conclusions: Our current research demonstrates that Lenalidomide when added to galiximab has augmented in vitro antitumor activity (i.e, antiproliferation and ADCC) and synergistic effects in vivo (i.e., prolongation of survival compared to either monotherapy). Our promising preclinical results of the unique combination of lenalidomide plus galiximab supports future evaluation of this doublet as a clinical trial. (Supported by USPHS grant PO1-CA103985 from the National Cancer Institute.)

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Correlations between baseline 18F-FDG PET tumour parameters and circulating DNA in diffuse large B cell lymphoma and Hodgkin lymphoma

EJNMMI Research ◽

10.1186/s13550-020-00717-y ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Pierre Decazes ◽

Vincent Camus ◽

Elodie Bohers ◽

Pierre-Julien Viailly ◽

Hervé Tilly ◽

...

Keyword(s):

B Cell ◽

Hodgkin Lymphoma ◽

Fdg Pet ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Circulating Dna ◽

Tumour Burden ◽

B Cell Malignancies ◽

Large B Cell Lymphoma ◽

Large B Cell

Abstract Background 18F-FDG PET/CT is a standard for many B cell malignancies, while blood DNA measurements are emerging tools. Our objective was to evaluate the correlations between baseline PET parameters and circulating DNA in diffuse large B cell lymphoma (DLBCL) and classical Hodgkin lymphoma (cHL). Methods Twenty-seven DLBCL and forty-eight cHL were prospectively included. Twelve PET parameters were analysed. Spearman’s correlations were used to compare PET parameters each other and to circulating cell-free DNA ([cfDNA]) and circulating tumour DNA ([ctDNA]). p values were controlled by Benjamini–Hochberg correction. Results Among the PET parameters, three different clusters for tumour burden, fragmentation/massiveness and dispersion parameters were observed. Some PET parameters were significantly correlated with blood DNA parameters, including the total metabolic tumour surface (TMTS) describing the tumour–host interface (e.g. ρ = 0.81 p < 0.001 for [ctDNA] of DLBLC), the tumour median distance between the periphery and the centroid (medPCD) describing the tumour’s massiveness (e.g. ρ = 0.81 p < 0.001 for [ctDNA] of DLBLC) and the volume of the bounding box including tumours (TumBB) describing the disease’s dispersion (e.g. ρ = 0.83 p < 0.001 for [ctDNA] of DLBLC). Conclusions Some PET parameters describing tumour burden, fragmentation/massiveness and dispersion are significantly correlated with circulating DNA parameters of DLBCL and cHL patients. These results could help to understand the pathophysiology of B cell malignancies.

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MicroRNA-181a Inhibits Activated B-Cell-Like Diffuse Large B-Cell Lymphoma Progression by Repressing CARD11

Journal of Oncology ◽

10.1155/2019/9832956 ◽

2019 ◽

Vol 2019 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Danxia Zhu ◽

Cheng Fang ◽

Wenting He ◽

Chen Wu ◽

Xiaodong Li ◽

...

Keyword(s):

B Cell ◽

Binding Site ◽

Cell Lines ◽

Expression Vector ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Wild Type ◽

Protein Levels ◽

Large B Cell Lymphoma ◽

Large B Cell

We investigated the role of miR-181a in diffuse large B-cell lymphoma (DLBCL) and its potential target genes. miR-181a levels were lower in activated B-cell- (ABC-) like DLBCL cells than that in germinal center B-cell- (GCB-) like DLBCL cells. Overexpression of miR-181a in ABC-like DLBCL cell lines (OCI-LY10 and U2932) resulted in G0/G1 cell cycle arrest, increased apoptosis, and decreased invasiveness. miRNA target prediction programs (miRanda, TargetScan, and miRDB) identified caspase recruitment domain-containing protein 11 (CARD11) as a putative miR-181a target. CARD11 mRNA and protein levels were higher in the ABC-like DLBCL than that in GCB-like DLBCL. Moreover, CARD11 mRNA and protein levels were downregulated in the OCI-LY10 and U2932 cell lines overexpressing miR-181a. Dual luciferase reporter assays confirmed the miR-181a binding site in the CARD11 3′UTR region. OCI-LY10 and U2932 cells transfected with a CARD11 expression vector encoding miR-181a with a mutated binding site showed higher CARD11 protein levels, cell viability, G2/M phase cells, and invasiveness compared to those transfected with a wild-type CARD11 expression vector. Nude mice xenografted with OCI-LY10 cells with overexpressed wild-type miR-181a generated smaller tumors compared to those with overexpressed mutated binding site of CARD11 3′UTR and miR-181a. These results indicate that miR-181a inhibits ABC-like DLBCL by repressing CARD11.

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