bluuebunny/crossref_metadata_2025_split huggingface dataset:
The BCL2 Targeted Deoxyribonucleic Acid Inhibitor (DNAi) PNT2258 Is Active in Patients with Relapsed or Refractory Non-Hodgkin’s Lymphoma
Authors: Harb Wael A. , Lakhani Nehal , Logsdon Ana , Steigelman Mary , Smith-Green Heidi , Gaylor Shari , Rodrigueza Wendi , Woolliscroft Michael , Sooch Mina , Messmann Richard A. , Al-Katib Ayad 
Published: October 2019

Abstract
Background: PNT2258 is a 24-base, single-stranded liposome-encapsulated DNA oligonucleotide that hybridizes to regions of the BCL2 gene, modulating its function. As previously reported, PNT2258 demonstrates antitumor activity in patients with advanced-stage NHL. The updated clinical data that are provided in this abstract (as of 07/25/14) now include anti-tumor activity in patients with DLBCL and Richter’s syndrome.
Methods: All patients had CT-measureable, FDG-PET positive disease with progression prior to study entry, and had previously received rituximab and combination cytotoxic chemotherapy. Refractory tumor status or concomitant medical issues precluded the use of additional cytotoxic therapy in all patients included in the study. PNT2258 induction cycles were administered as a 3-hour IV infusion at 120 mg/m2 days 1-5 of each 21-day cycle for up to 8 cycles. Response evaluation occurred at the end of cycles 2, 6 and 8. Following induction, patients could receive maintenance dosing at 100 mg/m2days 1-2 of each 28-day cycle until PD.
Results: Thirteen patients (median age 65; 8 males, 5 females; ECOG PS 0/1/2: 3/9/1) received all scheduled doses of PNT2258. Regardless of attribution, there were no cycle 1, grade 3/4 AEs. The most common grade 1/2 AEs in cycle 1 included chills, low-grade fever, transient nausea, and tumor or back pain (n=5 for each). There was no evidence of tumor lysis syndrome, febrile neutropenia or significant GI toxicity. Of 4 patients with DLBCL, including 1 with Richter’s syndrome and 1 with Burkitt-like histology, PFS was 9.2 months, range 5.5-12.3 months as of 7/25/14. Best response data (CR/PR/SD/PD) for the DLBCL group was 2/1/1/0. Additionally, in 5 FL patients, best response was 1/1/3/0. Five patients, including 3 DLBCL and 2 FL remain on study as of 7/25/14 receiving maintenance treatment with PNT2258.
Conclusions: PNT2258 is well tolerated and can be administered over prolonged periods of time. Durable single agent activity was previously reported in patients with FL, and has now been observed in patients with aggressive DLBCL. Studies in DLBCL and Richter’s syndrome are being implemented and updated information will be provided at the time of the meeting. Clinical trial information: NCT01733238.

Disclosures
Gaylor: ProNAi Therapeutics: Employment. Rodrigueza:ProNAi Therapeutics: Employment. Woolliscroft:ProNAi Therapeutics: Employment. Sooch:ProNAi Therapeutics: Employment. Messmann:ProNAi Therapeutics: Employment.
Rituximab, Age and High Dose Therapy Followed By Autologus Stem Cell Transplantation Are Independent Prognostic Factors for Survival in the First Line Treatment of Primary CNS-Lymphoma
Authors: Madle Michael , Herth Isabelle , Lehners Nicola , Schwarzbich Mark-Alexander , Wuchter Patrick , Egerer Gerlinde , Ho Anthony D. , Witzens-Harig Mathias 
Published: October 2019

Abstract
Introduction: For patients with diffuse large B-cell lymphoma without the involvement of the CNS, the addition of rituximab to standard chemotherapy has significantly improved survival. Thus far, there have been no prospective randomized trials evaluating the impact of rituximab as part of the primary treatment of PCNSL.
Methods: In this single-center, retrospective analysis, a total of 79 PCNSL-patients treated in our institution between 2000 and 2011 were included. Beside firstline chemotherapy with or without rituximab, we evaluated the impact of age (≤/> 60 years), autologous stem cell transplantation (ASCT +/-) and other factors upon overall survival (OS) and progression-free survival (PFS).
Results: In patients treated with rituximab (n=27), 3-year OS was 77.8% (95%-CI:62-93%). In contrast, in patients treated without rituximab (n=52), 3-year OS was only 39.9% (CI:27-53%, Figure). The difference in OS was significant in the univariate (p=0.002) as well as the multivariate analysis (p=0.049, Hazard ratio (HR)=0.248). In the rituximab group, 80.8% were free of progression at the date of analysis (median not reached), whereas median PFS in the group without rituximab was only 17 months (CI:8-26), (p=0.001). Patients ≤ 60 years of age (n=28) had a 3-year OS of 78.2% (CI:63-94%) and a median PFS of 75 months, in patients > 60 years (n=51) 3-year OS was 38.7% (CI:25-52%) and median PFS was 39 months (CI:6-72). Patients who received high dose therapy and autologous stem cell transplantation (ASCT) had a 3-year OS of 85.2% (CI:72-99%) and 65.1% were alive up to the time of analysis (range 9-131 months). Without ASCT median OS was only 16 months (CI:11-21) and 3-year OS was 35.2% (CI:22-48%). Age and ASCT were significantly associated with better OS in univariate (p=0.002 and p=0.001) as well in multivariate analysis (p=0.004, HR=0.023 and p=0.001, HR=0.014).
Conclusion: Rituximab treatment, ASCT and age are independent prognostic factors for overall survival in the first line treatment of PCNSL.
Figure 1 Figure 1.

Disclosures
Wuchter: Sanofi: Honoraria; ETICHO: Consultancy, Honoraria.
Follicular Lymphoma Survival in Sweden in the Rituximab Era
Authors: Junlén Henna Riikka , Peterson Stefan , Kimby Eva , Lockmer Sandra , Linden Ola , Nilsson-Ehle Herman , Erlanson Martin , Hagberg Hans , Rådlund Anders , Hagberg Oskar , Wahlin Björn E 
Published: October 2019

Abstract
Introduction
Follicular lymphoma (FL) is the most common indolent lymphoma, with 200-250 cases in Sweden per year. FL is considered incurable, because conventional therapy only leads to temporary remissions of varying length. Studies indicate that survival for patients with FL has improved since the introduction of rituximab. In Sweden first-line rituximab was gradually introduced between 2003 and 2007 and the speed of rituximab adoption varied geographically. In the publication of the first national Swedish guidelines for FL in November 2007, rituximab was recommended as first-line therapy and as maintenance after first relapse.
Methods
We conducted a nationwide, population-based registry study to assess patterns of FL survival before and after the introduction of first-line rituximab and to identify prognostic factors in the rituximab era. In the Swedish Lymphoma Registry (SLR) 2641 patients were identified with a FL diagnosed between January 1, 2000 and December 31, 2010. Clinical data, including first-line treatment were collected from the SLR. Overall survival (OS) and relative survival (RS) were estimated by calendar period (the pre-rituximab era 2000-2002, the transitional period 2003-2007 and the established period 2008-2010) and national health care region.
Results
First-line rituximab use increased gradually during the study period (2000-2002, 13%; 2003-2007, 61%; 2008-2010, 86%). During the transitional period, one health care region incorporated first-line rituximab quickly (92% first-line rituximab use) and one slowly (31%), whereas the remaining regions showed a moderate use of first-line rituximab (in total, 62%). OS and RS improved incrementally with each calendar period (P < 0.00005) (Figure 1A). Survival differed, during the transitional period, between regions with different rituximab use (OS, P = 0.039) (Figure 1B). After the national guidelines were published, no regional differences in use of first-line rituximab remained (P = 0.91). The improvement in OS and RS during the study period was greater in women than in men (Figure 1C). Survival improved in all age groups, but most in elderly patients (Figure 2).
Conclusions
The policy of first-line rituximab has resulted in a nationwide improvement in FL survival in Sweden. Women and elderly patients have benefited the most from rituximab.
Figure 1. Cumulative relative survival of follicular lymphoma by (A) calendar period of diagnosis, (B) regional adoption of rituximab during 2003-2007, and (C) calendar period of diagnosis and gender. Figure 1. Cumulative relative survival of follicular lymphoma by (A) calendar period of diagnosis, (B) regional adoption of rituximab during 2003-2007, and (C) calendar period of diagnosis and gender.
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Figure 2 Figure 2.

Disclosures
No relevant conflicts of interest to declare.
NFATc1 Is Transcriptionally Activated in Chronic Lymphocytic Leukemia (CLL) By Promotor DNA-Hypomethylation Which Correlates with in-Vitro Vulnerability to Calcineurin Inhibitors
Authors: Wolf Christine , Filarsky Katharina , Garding Angela , Claus Rainer , Oakes Christopher C , Idler Irina , Plass Christoph , Lichter Peter , Doehner Hartmut , Stilgenbauer Stephan , Mertens Daniel 
Published: October 2019

Abstract
Chronic lymphocytic leukemia (CLL), the most frequent adult leukemia in Western countries, is characterized by progressive accumulation of mature, monoclonal B lymphocytes in blood, bone marrow, and lymphoid tissues. In the pathogenesis and treatment of CLL, B cell receptor (BCR) signaling plays a crucial role, and aberrations in downstream pathways that become activated in CLL need to be better defined. One downstream target of BCR signaling is NFATc1, a transcription factor with a high oncogenic and transforming potential. Employing a genome-wide comparative DNA methylation analysis the NFATc1 5’ region was identified to be DNA hypomethylated in CLL patient samples. The pilot series comprised CD19+ B-cells from 13 CLL patients without previous treatment (9 with mutated IGHV using a 98% similarity cut-off) and CD19+ B-cells from 6 age-matched healthy donors. DNA methylation was relatively quantified using methyl-CpG immunoprecipitation (MCIp) on custom oligonucleotide promoter arrays covering -8kbp to +2.5kbp of all human transcription start sites. In total 1944 genes showed significant DNA hypomethylation in CLL cells compared to non-malignant B-cells (p<0.05), with NFATc1 ranking at position 36. DNA hypomethylation of the NFATc1 promoter was validated in peripheral blood CD19+ B-cells from 76 CLL patients without previous treatment compared to 14 additional age-matched healthy donors using mass spectrometry based quantitative high resolution measurement of DNA methylation (MASSARRAY). These findings suggest activation of the NFATc1 gene in CLL cells. In fact, NFATc1 mRNA was significantly upregulated in CLL samples compared to non-malignant B-cells, and expression correlated highly significantly with DNA methylation (r=-0.66, p<0.001). Inhibition of NFATc1 in primary CLL cells from patients (n=5) using calcineurin inhibitors like CsA and FK-506 induced significant apoptosis in primary CLL cells in-vitro even when protected by co-culture with bone-marrow stromal cells, suggesting a dependence of CLL cells on the activity of NFATc1. For a mechanistic insight into NFATc1 signal transduction in primary CLL cells and to allow modeling of the signaling network, a panel of phospho-specific antibodies is currently used in flow cytometry. This allows to monitor multiple components of signaling pathways in parallel with high time resolution to understand the impact of NFATc1 upregulation on the cellular signaling networks. The approach will shed further light on how CLL cell survival is regulated how this pathway can be targeted in CLL.

Disclosures
No relevant conflicts of interest to declare.
EZH2 Inhibition and the Combination with Proteasome Inhibition Are Novel Potential Strategies for the Treatment of Multiple Myeloma
Authors: Rizq Ola , Mimura Naoya , Koide Shuhei , Ma Anqi , Jin Jian , Iseki Tohru , Nakaseko Chiaki , Iwama Atsushi 
Published: October 2019

Abstract
PRC2 catalytic component EZH2 is a methyltransferase which induces trimethylation of histone H3 at lysine 27 (H3K27me3) to repress the transcription of target genes and promotes tumor growth in some cancers. Previous reports demonstrated that EZH2 is overexpressed in multiple myeloma (MM) cells and its expression level correlates with the progression of the disease, suggesting that EZH2 acts as an oncogene in MM. However, the detailed function of EZH2 in MM has not yet been elucidated. Therefore in this study, we examined the impact of EZH2 inhibition on MM cells and evaluated the preclinical efficacy of EZH2 inhibitors in the treatment of MM. We first examined the biologic impact of EZH2 knockdown using lentiviral shRNA and observed significant growth inhibition in knockdown cells. We next examined the effect of EZH2 inhibition using a novel selective EZH2 inhibitor UNC1999 (ACS Chem Biol. 2013). UNC1999 induced significant growth inhibition in MM cell lines such as MM.1S, H929, RPMI8226, and DOX40. We confirmed that UNC1999 reduced the levels of H3K27me3 in dose- and time-dependent manners in western blotting. Importantly, UNC1999 in combination with bortezomib demonstrated synergistic cytotoxicity analyzed by MTS assay in these cell lines. Even in the presence of conditioned media derived from bone marrow stromal cells, UNC1999 enhanced bortezomib-induced cytotoxicity in MM.1S cells. Similar synergistic effects were also observed in the combination of another EZH2 inhibitor GSK126 with bortezomib or in the combination of UNC1999 with another proteasome inhibitor carfilzomib. Moreover, apoptosis induced by bortezomib was enhanced by UNC1999 as evidenced by increased annexin V-positive cells in flow cytometric analysis. Apoptosis was further confirmed by cleavage of caspase-8, -9, -3 and PARP in western blotting. To assess the biologic impact of UNC1999 alone or in combination with bortezomib on MM cells, we performed gene set enrichment analysis using our microarray data. We found significantly positive enrichment of polycomb target genes in both UNC1999- and combination- treated MM.1S cells, and also in bortezomib-treated cells. Of interest, we observed that bortezomib alone downregulated EZH2 mRNA and EZH2 protein, which was enhanced by UNC1999, suggesting one mechanism of the synergistic effects of this combination. In order to examine the anti-MM activities of UNC1999, we used the MM.1S subcutaneous xenograft model in NOG mice. Intraperitoneal dosing of UNC1999 (25mg/kg) twice a week for three weeks significantly reduced the MM tumor growth in comparison to the control group (at day 22, p=0.04). In conclusion, these data indicate that selective EZH2 inhibition and the combination with proteasome inhibition are potential novel therapeutic options in MM.

Disclosures
No relevant conflicts of interest to declare.
Role of Erythroferrone in the Pathophysiology of Common Anemias
Authors: Kautz Leon , Jung Grace , Nemeth Elizabeta , Ganz Tomas 
Published: October 2019

Abstract
Introduction: We recently described the erythroid hormone erythroferrone (ERFE) as a critical regulator of hepcidin production during increased erythropoietic activity. Both anemia of inflammation (AI) and beta-thalassemia are associated with elevated levels of erythropoietin, the signal directing ERFE expression, suggesting that ERFE may play a role in the pathogenesis of these disorders. Indeed, Erfe mRNA expression was highly increased in the bone marrow and the spleen of a mouse model of β-thalassemia Hbbth3/+ (Th3/+). We therefore investigated the role of erythroferrone in hepcidin regulation in anemia of inflammation and β-thalassemia. We report that ERFE contributes to the recovery from anemia of inflammation and may be a hepcidin-suppressive factor responsible for iron accumulation in thalassemia.
Methods: We explored the role of ERFE in anemia of inflammation using the heat-killed Brucellaabortus (BA) mouse model of AI. Hepcidin regulation and recovery from anemia was compared between wild-type and Erfe-deficient mice. To determine whether ERFE is the hepcidin-suppressive factor in thalassemia, we generated Erfe-/-/Th3/+ double mutant mice and compared them to their littermate WT, Erfe-/- and Th3/+ mice. Mice were compared at 3, 6 and 12 weeks of age.
Results: Compared to wild-type mice, Erfe-deficient mice did not appropriately decrease hepcidin production during recovery from AI and exhibited greater severity of anemia (figure). These data suggest that compensatory hepcidin suppression during the recovery phase of AI is mediated by ERFE. However, in mice this defect was partly compensated by prolonged stimulation of erythropoiesis and higher reticulocytosis resulting in Erfe-deficient mice reaching wild-type hemoglobin levels by day 28 (figure).
Figure 1 Figure 1.
At the other end of the spectrum, we showed that thalassemic mice exhibited greatly increased expression of Erfe mRNA in the bone marrow and the spleen. Ablation of Erfe in Th3/+ mice restored normal hepcidin levels and significantly reduced serum iron concentration and hepatic iron overload at 6 weeks of age. Comparison of Erfe-/- / Th3/+ and Th3/+ mice at 3, 6 and 12 weeks of age did not show any difference in the severity of the anemia in absence of ERFE suggesting that ERFE regulates hepcidin expression but its absence did not cause iron restriction and did not ameliorate ineffective erythropoiesis.
Conclusion: ERFE is a critical regulator of hepcidin expression during recovery from AI and ERFE agonists may alleviate iron restriction in AI. ERFE may also be the factor responsible for hepcidin suppression and secondary iron overload in β-thalassemia.

Disclosures
Ganz: Intrinsic LifeSciences: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees; Keryx Pharma: Consultancy; Merganser Biotech: Consultancy, Equity Ownership.
Molecular Signatures of Tumor-Initiating Cells Unveil Wnt Pathway As a Therapeutic Target in Mantle Cell Lymphoma
Authors: Samaniego Felipe , Sehgal Lalit , Braun Frank K , Berkova Zuzana , Romaguera Jorge E. , Wang Michael , Rodriguez M. Alma , Neelapu Sattva S , Mathur Rohit 
Published: October 2019

Abstract
Introduction
Mantle cell lymphoma (MCL) is an aggressive and incurable form of non-Hodgkin’s lymphoma. Despite initial responses to intense chemotherapy, up to 50% of cases of MCL relapse, often in a chemoresistant form. We hypothesized that the recently identified MCL-initiating cells (MCL-ICs) are the main reason for relapse and chemoresistance of MCL.
Methods
We isolated MCL-ICs from primary MCL cells on the basis of a defined marker expression pattern; CD34-CD3-CD45+CD19-. The MCL-ICs, MCL-non-ICs, and peripheral blood lymphocytes from healthy donors were analyzed for gene expression using the Arraystar platform. The differences in mRNA levels of genes of interest were confirmed by quantitative RT-PCR. The prominent differentially expressed transcripts were analyzed using the Ingenuity Platform. Primary MCL cells were co-culture with mesenchymal stem cells to assess the effects of chemotherapeutic agents such as vincristine, doxorubicin and the newly approved Burton tyrosine kinase inhibitor ibrutinib, and Wnt signaling inhibitors.
Results
Approximately 1% of primary MCL cells are MCL-ICs and they can be maintained in co-culture with mesenchymal stem cells. Comparison of gene expression profiles of MCL-ICs and MCL-non-ICs revealed activation of stem cell-specific pathways in MCL-ICs by expression of Wnt, Notch, and Hedgehog and enhanced expression of Nanog, Oct4, KLF4, ADH1, MT1b and ABCC3. Gene expression microarray data and RT-PCR data suggested predominant activation of the Wnt/Frizzled pathway. Indeed, MCL-ICs were particularly sensitive to Wnt pathway inhibitors. Targeting Wnt-dependent β-catenin‒TCF4 interaction with CCT036477, iCRT14, or PKF118-310 preferentially eliminated the MCL-ICs, reduced the expression of stem cell transcription factors (Myc, Nanog, Oct4, Klf4), and sensitized MCL cells to vincristine, doxorubicin, and ibrutinib. Interestingly, while vincristine, doxorubicin or ibrutinib did kill MCL cells, they did not reduce the percentage of MCL-ICs in treated co-culture.
Conclusion
MCL-ICs are present in primary MCL isolates and they show gene expression pattern of chemoresistant, stem cell-like cells with predominant activation of Wnt signaling. In order to produce durable remissions in MCL patients, treatment strategies should be directed to target MCL-ICs.

Disclosures
Wang: Pharmacyclics, Janssen: Honoraria, Research Funding.
The Inhibitor of NF-KB Kinase, IKKβ Regulates the Transcriptional Activity of GLI1 By Blocking Its Proteasomal Degradation
Authors: Agarwal Nitin Kumar , Kim Chae Hwa , Kunkalla Kranthi , Lossos Izidore S. , Vega Francisco 
Published: October 2019

Abstract
GLI1 is a Hedgehog (Hh) related transcription factor originally discovered as an amplified product in gliomas. Inappropriate activation of the GLI1 has been shown in many cancers including diffuse large B cell lymphoma (DLBCL). We previously showed that GLI1 mediated canonical Hh signaling is constitutive active in DLBCL and contributes to cell survival, proliferation and enhances chemotolerance. Although the importance of GLI1 in tumor development is well recognized, the molecular mechanisms controlling the transcriptional activity of GLI1 are poorly characterized.
To identify regulatory components that participate in the transcriptional activity of GLI1, we explored GLI1 putative interacting proteins by liquid chromatography tandem mass spectrometry following immunoprecipitation of endogenous GLI1. We detected that the inhibitor of NF-KB kinase, IKKβ, is one of the proteins associated with GLI1 transcription factor. Here we investigate the regulatory role of IKKβ in the transcriptional activity of GLI1. We show that IKKβ regulates the transcriptional activity of GLI1 by phosphorylating GLI1 in C-terminal region and modulating its protein stability. Short stimulation of SUDHL4 and DOHH2 cells with TNF-α (20ng/mL) resulted in increased GLI1 protein levels. Similar results were observed in 293T cells transiently transfected with GLI1 and IKKβ kinase constructs. Moreover, silencing of IKKβ using siRNA and shRNAs led to decreased GLI1 protein levels and its transcriptional activity in DLBCL cell lines with constitutive activation of the NF-KB.
Next, we characterized nine probable IKKβ dependent GLI1 phosphorylation sites (S543-S548, S1070, S1071 and S1074 identified by nanospray ion trap mass spectrometry) using mutational and deletions studies. We show that IKKβ phosphorylates GLI1 at Thr1074 and decreases binding between GLI1 and HECT-type E3 ubiquitin ligase (ITCH) resulting in reduced GLI1 polyubiquitination and degradation. Point mutation of Threonine 1074 to Alanine prevents IKKβ-mediated GLI1 phosphorylation and facilitates GLI1-ITCH interaction, polyubiquitination and degradation of GLI1 in the proteasome.
Collectively, our data links IKKβ-mediated NF-kB signaling to the transcriptional activity of GLI1 and illustrates a novel cross talk between these two pathways. This is of clinical interest because activation of the NF-kB pathway is frequent in DLBCL and the connection between Hh and NF-kB pathways may open novel therapeutic avenues for DLBCL.

Disclosures
No relevant conflicts of interest to declare.