bluuebunny/crossref_metadata_2025_split huggingface dataset:
SNX 2112, An Oral Hsp-90 Inhibitor Exerts Antiproliferative Effects in Combination with Bortezomib and Rituximab in Rituximab Resistant Non-Hodgkin's Lymphoma.
Authors: Reddy Nishitha , Hicks Donna , Jagasia Madan , Amiri Kathy 
Published: October 2019

Abstract
Abstract 3733
Poster Board III-669

Background
Heat Shock protein 90 (Hsp90) is a chaperone that plays an important role in maintaining the stability of various other client proteins involved in cell survival and proliferation. Few client proteins that are dependent on Hsp-90 include Akt, Raf, ERK and NF-κB. Hsp-90 is upregulated in several malignancies and thus serves as an important therapeutic target. SNX 2112 (PF-04928473) is an oral heat shock protein inhibitor that is currently under phase-I clinical trials.

Methods and Results
We examined the effect of increasing concentrations of SNX 2112 in combination with other active targeted agents in Non-Hodgkin's lymphoma. In our current work, we exposed rituximab sensitive (RSCL) and rituximab resistant cell lines (RRCL) to varying concentrations of SNX-2112 (500nM, 1μM, μM, 10μM and 20 μM) along with fixed doses of rituximab (10μg/ml) as well as bortezomib (500nM) and performed growth inhibition assays. The growth inhibitory effects of the various agents in Lymphoma cell lines were assessed by measuring 3-2, 5-diphenyl tetrasodium bromide (MTT) dye absorbance. Cells were incubated for 24 hrs and 48hrs. MTT was added to each well the last 4 hrs. Absorbance was measured using spectrophotometer. Accordingly, immunoblotting was performed using lymphoma cell lysates. The whole cell lysates were subjected to polyacrylamide gel electrophoresis and nitrocellulose membranes were immunoblotted with phosphorylated Hsp-90 and α-tubulin. In parental raji cell lines, at 48 hrs, and a dose of 10 μM, SNX 2112 demonstrated potent anti-proliferative effects when combined with bortezomib and rituximab 0.23 OD vs. rituximab alone 1.06 OD (p=0.010) 95% CI; 0.47, 1.18. In rituximab resistant cell lines, SNX 2112 with rituximab and bortezomib demonstrated significant antiproliferative effects 0.199 OD vs. rituximab alone 0.86 OD (p=0.023) 95% CI 0.22-1.11. As a single agent neither rituximab or bortezomib nor the combination of these agents was significant in inhibiting cell growth in rituximab resistant cell lines. Cytotoxicity was more significant in the rituximab resistant cell lines that rituximab sensitive raji cell lines. Immunoblotting performed using phosphorylated hsp-90 also demonstrated significant inhibition when treated with SNX 2112.

Conclusions
Our current research demonstrates that SNX 2112 when added to bortezomib and rituximab has augmented anti-tumor activity (ie, antiproliferation) and demonstrates additive effects. Further in vivo studies are underway to demonstrate this effect. Our promising pre-clinical results of this unique combination support further evaluation in a clinical trial in patients with relapsed Non-Hodgkin's lymphoma.

Disclosures:
No relevant conflicts of interest to declare.
Treatment Model of CNS Lymphoma Using Complement-Dependent Cytotoxicity Induced by Rituximab and Autoserum.
Authors: Miyake Yasuyuki , Okoshi Yasushi , Machino Takayuki , Chiba Shigeru 
Published: October 2019

Abstract
Abstract 3739
Poster Board III-675

Background
Primary central nervous system lymphoma (PCNSL) is almost exclusively CD20-positive non-Hodgkin lymphoma (NHL). Although rituximab (R) is widely used for CD20-positive NHL, it is not considered to reach brain lesions effectively beyond the blood brain barrier. Intraventricule administration (ivt) of R is reported to be effective in meningeal lymphoma but the effect on lesions in the brain parenchyma seems to be limited. Recently, a case of refractory PCNSL that was successfully treated with ivt of R with autologous serum was reported (Takami A, et al. Cancer Science, 2006). Because the cerebrospinal fluid does not contain complements which exists in the serum, induction of complement-dependent cytotoxicity by ivt of R plus autoserum was speculated. To investigate this effect, we developed an animal treatment model of CNSL.

Materials and methods
Raji, CD20-positive Burkitt lymphoma cell line, was inoculated into the deep frontal lobe of the brain of 8-week old F344 (nru-/nru-) nude rats, using brain stereotaxic apparatus. At the same time, a cannula was placed into the ipsilateral lateral ventricle. After several days, R or control immunoglobulin (cIg), plus human serum or saline, was administrated into the lateral ventricle.

Results
The brain was extracted 24 hours after the last administration and frozen section was made. Human CD20-positive Raji cell tumor was also positively stained with FITC-conjugated anti-human IgG antibody when R but not cIg was administrated. Consequently, R in the lateral ventricle was considered to penetrate ependymal cells and brain parenchyma, and bound to lymphoma cells. Next, these rats were treated with ivt of R plus serum (R + Serum), cIg plus serum (cIg + Serum), or R plus saline (R + saline). These were administrated once a day from day 5 to day 9 after inoculation of Raji, and then survival was monitored. When an obvious weakness, such as marked and consecutively loss of activity or weight, was observed, these rats were euthanized and this is defined as dead day. In each case, the brain was extirpated and examined whether lymphoma existed or not. Death without lymphoma or from technical problem was excluded from the analysis. Survival of each group was analyzed by Kaplan-Meier method and log-lank test. R + Serum group had longer survival than cIg + Serum (p = 0.049). Long-term survivors were only seen in R + Serum and this group seemed to be superior to R + saline but statistical difference was not detected (p = 0.083). There were no difference between cIg + Serum and R + saline (p =0.382) and neither group had long-term survivor.

Conclusion
The possibility of novel treatment of CNSL with ivt of R and autoserum was shown in the rat CNSL model. To confirm this approach, clinical trials are warranted.

Disclosures:
No relevant conflicts of interest to declare.
A Phase II Trial of R-FM (Rituximab, Fludarabine and Mitoxantrone) Chemotherapy Followed by Yttrium 90 (90Y) Ibritumomab Tiuxetan (90Y-IT) for Untreated Follicular Lymphoma (FL) Patients.
Authors: Zinzani Pier Luigi , Tani Monica , Broccoli Alessandro , Derenzini Enrico , Pulsoni Alessandro , De Renzo Amalia , Fanti Stefano , Fina Mariapaola , Gandolfi Letizia , Pellegrini Cinzia , Stefoni Vittorio , Baccarani Michele 
Published: October 2019

Abstract
Abstract 3743
Poster Board III-679

Introduction
In 2008 we published a multicenter non-randomized phase II trial of fludarabine and mitoxantrone plus 90Y-IT in untreated patients with FL. By the end of the entire treatment regimen 95% of the patients achieved complete remission (CR). With a median follow-up of 30 months, 3-year PFS was estimated to be 76% and 3-year OS 100%. On the basis of these results we are currently conducting a prospective, multicenter, non-randomized, phase II study of R-FM followed by 90Y-IT in untreated patients with FL in which the number of fludarabine and mitoxantrone cycles has been decreased to four and in which rituximab is administered before each cycle. The rationale of the trial is to use different forms of treatment and to reduce the use of conventional chemotherapy and its related toxic effects.

Patients and Methods
Patients eligibility is represented by: age more than 18, stage II-IV, FL grade I-II, WHO performance status 0-2. Patients are treated with standard FM chemotherapy plus rituximab every 28 days for 4 cycles. Patients are restaged 4 to 8 weeks after completion of immunochemotherapy and those achieving at least a partial response are eligible for 90Y-IT. All patients receive a single dose of 90Y-IT 14,8 MBq/kg. At the time of the analysis we enrolled 55 patients. 25 patients were male and 30 female; the median age was 56 years (range 26-84); 12 patients were stage II, 13 stage III and 30 stage IV; 11 patients had a bulky disease. 52 patients completed the induction chemotherapy, all except 5 were eligible for the consolidation treatment with 90Y-IT and 44 patients were restaged after the entire treatment regimen.

Results
After the R-FM chemotherapy, the overall response rate was 92.3% (48/52) including 39 (75%) CR and 9 (17.3%) partial remissions (PR). Time to event analyses, including TTP and duration of response are pending further follow-up. Treatment was well tolerated grade 3-4 haematologic AEs (mostly neutropenia) were seen in 50% of the patients. Among the 44 patients (9 PR and 35 CR) subsequentially treated with 90Y-IT and reassessed for the response, 8/9 (88.9%) PR patients improved their remission status from PR to CR. 90Y-IT toxicity included mostly grade 3-4 neutropenia and thrombocytopenia and was comparable to the literature data.

Conclusions
These preliminary data indicate that radioimmunotherapy appears highly effective and feasible as “consolidation” after short (4 cycles) immunochemotherapy in FL patients, improving quality of response without any cumulative toxicity.

Disclosures:
No relevant conflicts of interest to declare.
Ad-ISF35-Transduced Autologous Cells Promote in Vitro and In Vivo Chemosensitization to FCR in Patients with Del(17p) / P53 Defective Chronic Lymphocytic Leukemia.
Authors: Castro Januario E. , Melo-Cardenas Johanna , Sandoval-Sus Jose , Urquiza Mauricio , Prussak Charles , Kipps Thomas J. 
Published: October 2019

Abstract
Abstract 376
Chronic lymphocytic leukemia (CLL) cells that have del(17p) typically have loss of functional P53, rendering these cells refractory to standard chemotherapeutic agents, which require activation of P53 for their cytotoxic activity. However, del(17p) CLL cells co-cultured in vitro with cells transduced to express the CD40L (CD154) activate another member of the P53 family of proteins, namely P73, which, like P53, can induce transcription of death receptors and pro-apoptotic proteins and sensitize cells to the cytotoxic activity of “P53-dependent” drugs, such as Fludarabine (F-ara-A). Moreover, transduction of del(17p) CLL cells with a replication-defective adenovirus (Ad) encoding recombinant CD154 (Ad-ISF35) can induce such changes in both transduced and bystander CLL cells in vitro. To examine whether a similar activity could be obtained in vivo, we are conducting a phase Ib clinical study in subjects with high-risk CLL who are refractory to fludarabine or have evidence of del(17p). In this study, subjects receive three IV doses of 3×108 autologous CLL cells that had been transduced ex vivo with Ad-ISF35 and two weeks latter they are treated with a truncated chemoimmunotherapy regimen involving 3 monthly courses of fludarabine, cyclophosphamide and rituximab (FCR). P53-defective CLL cells from treated patients were initially resistant to F-ara-A induced apoptosis with IC50 > 10μM prior to treatment. CLL cells collected from patients ≥ 24 hours after the first infusion of autologous Ad ISF35-trasduced CLL cells became sensitive to the cytotoxic effects of F-ara-A, with IC50 0.3-1 μM. Enhanced sensitivity to F-ara-A was associated with induced expression of Bid, DR5, CD95, and P73 by circulating non-transduced “bystander” CLL cells, an effect lasting ≥ 2 weeks following IV infusion. To date, two subjects have completed treatment and this has been well tolerated without serious adverse events. The most common adverse events have been transient fever, malaise and fatigue associated to infusion of Ad-ISF35 transduced cells and cytopenias after treatment with FCR. Both subjects have achieved a compete response, one of them without detectable minimal residual disease (MRD) by sensitive multiparameter flow cytometry of marrow mononuclear cells obtained 3 months following completion of treatment. Moreover, these patients have complete resolution of lymphadenopathy and organomegaly by exam and by whole body computed tomography. These results indicate that Ad-ISF35-cell-gene therapy can sensitize P53-deficient CLL to “P53-dependent” cytotoxic agents in vivo, allowing for effective treatment of patients who otherwise would be resistant to standard forms of therapy.

Disclosures:
Prussak: Memgen:
Employment.
OSU-DY7, a Novel D-Tyrosinol Derivative, Mediates Cytotoxicity in Chronic Lymphocytic Leukemia and Lymphoblastic Lymphoma through p38 Mitogen-Activated Protein Kinase Pathway.
Authors: Bai Li-Yuan , Ma Yihui , Kulp Samuel K , Wang Shu-Huei , Chiu Chang-Fang , Frissora Frank , Mo Xiaokui , Jarjoura David , Byrd John C. , Chen Ching-Shih , Muthusamy Natarajan 
Published: October 2019

Abstract
Abstract 3778
Poster Board III-714

Introduction
Chronic lymphocytic leukemia (CLL) is a most common form of adult leukemia with minimal treatment options. Drug resistance and associated immune deregulation limit use of current therapies, thus warranting need for alternative therapy development. Our laboratories recently identified a novel D-tyrosinol-derived compound targeting p38 MAPK pathway, OSU-DY7 [(R)-2-amino-3-(4-heptyloxy-phenyl)-propan-1-ol]. Here we demonstrate the efficacy of OSU-DY7 for lymphocytic cell lines and primary B cells from CLL patients.

Materials and Methods
We tested the pre-clinical efficacy of OSU-DY7 in primary CLL B cells and B cell lines representing CLL (MEC-1), ALL (697), and lymphoblastic lymphoma (Raji and Ramos).

Results
The cytotoxicity of OSU-DY7 was both dose- and time-dependent. The IC50 of OSU-DY7 at 24 hrs for MEC-1, 697, Raji, Ramos and primary B-CLL cells were 2.40 μM, 10.39 μM, 6.04 μM, 3.75 μM and 3.58 μM respectively. OSU-DY7 induced activation of caspase-3 and poly (adenocine diphosphate-ribose) polymerase (PARP) cleavage. Pancaspase inhibitor Z-VAD-FMK partially rescued OSU-DY7-induced cytotoxicity in CLL B cells and cell lines. Interestingly, OSU-DY7 mediated-cytotoxicity was associated with increased phosphorylation of p38 MAPK and its downstream target protein MAPKAPK2 in both lymphocytic cell lines and primary B-CLL cells. Compared with control group, the ratio of p-p38MAPK (Thr180Tyr182) versus p38MAPK in Raji cells treated with 2 μM, 4 μM and 8 μM of OSU-DY7 increased 2.2, 4.7 and 11 fold respectively (p<0.0001). Similar increase in p38MAPK phosphorylation was also observed in six CLL patient samples after treatment with OSU-DY7 2 μM, 4 μM and 8 μM for 24 hours (p=0.0004). Moreover, SB202190, a specific p38 MAPK inhibitor, decreased MAPKAPK2 protein level with concomitant rescue of the cells from the OSU-DY7 mediated-cytotoxicity. Thus pretreatment of Raji and primary CLL B cells with SB202190 reduced the OSU-DY7 induced cytotoxicity by 36.7% and 24.3% respectively by 24hrs (Raji:p<0.0001; primary CLL B cells: p<0.005 ). Furthermore, SB202190 rescued OSU-DY7 down regulated survivin protein by ∼2 fold (p<0.0001, n=3) and mRNA levels by 2.4 fold (95% CI: 1.56∼3.84 fold, p=0.0026, n=3) in Raji cells indicating a role for p38MAPK dependent regulation of survivin by OSU-DY7.

Conclusions
This study provides an evidence for a role of OSU-DY7 in p38 MAPK dependent activation and survivin down regulation associated with apoptosis in lymphocytic cells, thus warranting further development of this novel agent for alternative therapy for lymphocytic malignancies. [This work was supported by D. Warren Brown Foundation and Leukemia and Lymphoma Society]

Disclosures:
No relevant conflicts of interest to declare.
The PI3K Inhibitor GDC-0941 Synergizes with Standard of Care Therapies to Induce Growth Inhibition and Apoptosis of Multiple Myeloma Cells.
Authors: Munugalavadla Veerendra , Berry Leanne , Du Changchun , Mariathasan Sanjeev , Slaga Dion , Sun Laura , Chesi Marta , Bergsagel Peter Leif , Ebens Allen J. 
Published: October 2019

Abstract
Abstract 3788
Poster Board III-724
Multiple myeloma (MM) is a malignancy characterized by clonal expansion and accumulation of long-lived plasma cells within the bone marrow. Phosphatidylinositol 3' kinase (PI3K) -mediated signaling is frequently dysregulated in cancer and controls fundamental cellular functions such as cell migration, growth, survival and development of drug resistance in many cancers, including MM, and therefore represents an attractive therapeutic target. Here, we demonstrate in vitro, that a potent and selective pan-isoform PI3Kinhibitor, GDC-0941, modulates the expected pharmacodynamic markers, inhibits cell-cycle progression and induces apoptosis; overcomes resistance to apoptosis in MM cells conferred by IGF-1 and IL-6; and is additive or synergistic with current standard of care drugs including dexamethasone, melphalan, lenolidamide and bortezomib. In cell lines we find sensitivity to GDC-0941 is positively correlated with pathway activation as determined by phospho-AKT-specific flow-cytometry and Western-blot analysis. Preliminary results indicate apoptosis of MM cells is correlated with increased expression of the proapoptotic BH3-only protein BIM; mechanisms of increased apoptosis in MM will be further explored and an update presented. We further extend these in vitro findings to show that GDC-0941 has activity as a single agent in vivo and combines well with standard of care agents in several murine xenograft models to delay tumor progression and prolong survival. Our results suggest that GDC-0941 may combine well with existing therapies, providing a framework for the clinical use of this agent, and a rational approach to improving the efficacy of myeloma treatment.

Disclosures:
Munugalavadla: Genentech: Employment, Patents & Royalties. Berry:Genentech: Employment, Patents & Royalties. Du:Genentech, Inc.: Employment, Equity Ownership. Mariathasan:Genentech: Employment, Patents & Royalties. Slaga:Genentech: Employment, Patents & Royalties. Sun:Genentech Inc.: Employment. Chesi:Genentech, Inc.: Consultancy; Amgen: Consultancy; Celgene: Consultancy; Merck: Research Funding. Bergsagel:Genentech: Consultancy; Amgen: Consultancy; Celgene: Consultancy; Merck: Research Funding. Ebens:Genentech, Inc.: Employment, Equity Ownership, Patents & Royalties.
Patients with Del(5q) MDS Who Fail to Achieve Erythroid or Cytogenetic Remission After Treatment with Lenalidomide Have An Increased Risk for Clonal Evolution and AML Progression.
Authors: Göhring Gudrun , Giagounidis Aristoteles , Büsche Guntram , Kreipe Hans , Zimmermann Martin , Hellstrom-Lindberg Eva , Aul Carlo , Schlegelberger Brigitte 
Published: October 2019

Abstract
Abstract 3818
Poster Board III-754
Lenalidomide consistently induces transfusion independence and complete cytogenetic response in patients with low and intermediate risk myelodysplastic syndromes (MDS) with 5q deletion. Only limited information on long-term outcome of patients treated with this novel approach is currently available. We performed a long-term follow-up analysis of 42 patients with low or intermediate risk MDS and 5q deletion treated with lenalidomide. At a median follow-up of 40 months, 58% of the patients achieved an erythoid response and 48% a cytogenetic response. 36% of patients progressed into acute myeloid leukaemia (AML). Most of them (87%) acquired chromosome aberrations in addition to the 5q deletion, i.e. underwent clonal evolution during leukaemogenesis. There were no clinical, cytological or cytogenetic markers at study entry that allowed prediction of an increased risk of leukaemic transformation. However,erythroid and cytogenetic responders had a significantly decreased risk of progression to AML (p=0.001 and p=0.009, respectively) compared to non-responders. 3 and 5 years after study entry, the cumulative incidence of AML for patients with a cytogenetic response was 10% and 21%, respectively, and for patients without cytogenetic response, it was 46% and 60%. Patients with del(5q) MDS who fail to achieve erythroid or cytogenetic remission after treatment with lenalidomide have an increased risk for clonal evolution and AML progression. In refractory, or relapsing, patients, genetic instability and clonal evolution seem to be the driving forces of leukaemic transformation. Regular follow-up investigations of del(5q) MDS patients treated with lenalidomide may help identifying patients needing alternative treatment strategies.

Disclosures:
Göhring: Celgene Corp.: cytogenetic reference lab for Celgene MDS-004 study. Giagounidis:Novartis Pharma: Consultancy; Amgen: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Glaxo SmithKline: Consultancy; Johnson&Johnson: Consultancy; Celgene Corp.: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. Büsche:Celgene Corp.: histology reference lab for MDS-004 study. Kreipe:Celgene Corp.: histology reference lab for MDS-004 study. Hellstrom-Lindberg:Celgene Corp.: Research Funding. Schlegelberger:Celgene Corp.: cytogenetic reference lab for Celgene MDS-004 study.
Interleukin-6 (IL-6) Enhances C-MYC Translation IN MULTIPLE MYELOMA (MM) CELLS: ROLE of IL-6-INDUCED EFFECTS On the C-MYC RNA-Binding PROTEIN, HNRNPA1.
Authors: Shi Yijiang , Gera Joseph , Lichtenstein Alan 
Published: October 2019

Abstract
Abstract 3839
Poster Board III-775
Our previous work (Cancer Research 68:10215, 2008) demonstrated that IL-6 enhanced c-myc protein expression in MM cells and function of the RNA-binding protein, hnRNPA1 (A1), was required. This occurred by way of enhanced cap-independent translation mediated via the internal ribosome entry site (IRES) in the 5'UTR of the myc RNA. IRES-dependent translation is the fail safe mechanism for protein expression when cap-dependent translation is suppressed by mTOR inhibition (with curtailed RNA cap-ribosome binding) and is especially important when an mRNA leader is relatively long and highly structured, such that scanning ribosomes are unlikely to efficiently initiate translation. The IRES directly recruits the ribosome to within close proximity to the start codon, bypassing the need for cap binding and ribosome scanning. HNRNPA1 (A1) is a documented IRES trans-acting factor (ITAF) for the myc IRES, facilitating translation but how its effects are enhanced by IL-6 is unknown. HNRNPA1 is a shuttling protein which binds the myc RNA in the nucleus and transports it to the cytoplasm. Initial experiments demonstrated that IL-6 stimulation of ANBL-6 and OCI-My5 MM cell lines, as well as several primary MM specimens, significantly increased the cytoplasmic localization of A1. To test if this enhanced cytoplasmic localization was critical, we stably expressed a dominant negative (DN) A1 gene that is incapable of nuclear-to-cytoplasmic shuttling and which also prevents endogenous hnRNPA1 shuttling. The DN also prevented A1-mediated transport of the shuttling protein, FUS. Expression of the DN in ANBL-6 cells prevented IL-6-induced effects on myc expression and on ANBL-6 cell growth. We also tested if IL-6 treatment affected A1 binding to the myc RNA by immunoprecipitating A1 and performing real time PCR on the immunoprecipitate for myc RNA levels. A significant increase in A1-myc RNA binding was confirmed. Mass spectroscopy demonstrated that IL-6 induced phosphorylation of A1 in its RNA-binding domain, which possibly mediated the enhanced binding. These results demonstrate that, by enhanced binding of the myc ITAF, hnRNPA1, to the myc IRES, and by enhanced transport of the complex to the translationally active cytoplasmic subcellular site, IL-6 may stimulate c-myc translation and subsequent MM cell growth.

Disclosures:
No relevant conflicts of interest to declare.