bluuebunny/crossref_metadata_2025_split huggingface dataset:
Incidence and Outcomes of Myeloproliferative Neoplasms (MPN) in Adolescents and Young Adults (AYAs).
Authors: Pemmaraju Naveen , Kantarjian Hagop M , Cortes Jorge E. , Quintas-Cardama Alfonso , Pierce Sherry A. , Verstovsek Srdan 
Published: October 2019

Abstract
Abstract 2845

Background:
The MPNs are a family of chronic hematologic malignancies that typically affect pts ages 60–70s. However, there is growing awareness of hematologic malignancies developing in young pts, particularly a unique subset of pts known as AYAs. Little is known about incidence and outcomes of AYA pts with MPNs.

Objectives:
To determine the incidence and outcomes of MPN AYA pts.

Results:
We retrospectively reviewed charts of 1,616 MPN pts evaluated at our institution from 1986–2011. A total of 171 MPN pts (11%) were identified whose ages were 16–39, defined as AYA pts (per NCCN guideline recommendations on AYA cancers). Breakdown by MPN subtype: Essential Thrombocytosis, ET (n=78, 46% of AYA MPNs), Polycythemia Vera, PV (n=25, 15%), Myelofibrosis, MF(n=24, 14%), [3 post-ET, 1 post-PV, 20 primary MF], Hypereosinophilic syndrome, HES (n=22, 13%), and Systemic Mastocytosis, SM (n=22, 13%). Baseline characteristics of AYA MPN pts detailed in Table 1. Only significant difference among AYA and non-AYA pts in terms of baseline CBC parameters was WBC: 6.4 (0.4–108.4) in AYA MF vs 10.1 (0.6–361) in non-AYA MF, p=0.0049. Analysis of median overall survival (OS), by Kaplan-Meier method, compared by age group (AYA vs non-AYA pts) and broken down by MPN subtype, shown in Table 2.

Conclusion:
MPN AYA pts constitute 11% of MPN pts at our institution. Overall, their 5- and 10- yr OS were significantly better than their older counterparts in the 3 major MPN subtypes (ET, PV, MF) and trended towards better survival but not statistically significant in HES and SM. Among AYA MPN pts, female predominance was noted among ET and SM pts and only 1 transformation event was noted among all MPN AYA pts in this analysis.

Disclosures:
No relevant conflicts of interest to declare.
Signal Transducer and Activator of Transcription (STAT)-3 Induces the Expression of Microrna-155 in Chronic Lymphocytic Leukemia (CLL) Cells.
Authors: Li Ping , Grgurevic Srdana , Liu Zhiming , Harris David , Rozovski Uri , Keating Michael J. , Calin George , Estrov Zeev 
Published: October 2019

Abstract
Abstract 2885
MicroRNAs (miRs) are involved in the initiation, progression and dissemination of CLL cells (Calin GA, Croce CM. Blood 114:4761, 2009). Recent studies showed that high levels of miR-155, previously shown to regulate hematopoietic cell development, are expressed in CLL cells. Because transgenic miR-155 overexpression in the mouse stimulates B-cell proliferation, it is thought that miR-155 plays a role in the pathogenesis of CLL (Calin GA et al. N Engl J Med 353:1793, 2005). STAT3 is constitutively activated in CLL and induces the transcription of several STAT3-regulated genes. A recent study demonstrated that STAT3 activates miR-21 and miR-181b-1 (Iliopolus D. et al. Mol Cell 39:493, 2010). Therefor we wondered whether STAT3 enhances the expression of miR-155 in CLL cells. Because a sequence analysis revealed that the promoter of miRNA-155 harbors γ-interferon activation sequence-like elements typically activated by STAT3, we sought to determine whether STAT3 directly activates miR-155 expression. We generated truncated constructs of the miR-155 promoter, co-transfected them into MM1 cells together with STAT3 small interfering (si) RNA (siRNA), and assessed their luciferase activity. The luciferase activity data suggested that of the two putative STAT3 binding sites only one site is involved in STAT3 induced transcription because STATR3-siRNA reduced the activity of miRNA-155 promoter of constructs that harbor this site. To confirm these data we performed an electrophoretic mobility shift assay (EMSA) and chromatin immune-precipitation (ChIP). The EMSA confirmed that STAT3 bound the miR-155 promoter in fresh CLL cells, and ChIP confirmed that STAT3 bound one putative STAT3-binding site in the miR-155 promoter but not to the other, as demonstrated by the luciferase assay; STAT3 co-immuno-precipitated only one putative STAT3 binding region of miR-155 promoter and other STAT3-regulated genes. Finally, STAT3-small hairpin RNA (shRNA) downregulated miR-155 and other STAT3-regulated genes, suggesting that constitutively activated STAT3, binds miR-155 promoter and induces miR-155 transcription in CLL cells.

Disclosures:
Keating: Celgene Corporation: Consultancy, Research Funding; Roche: Consultancy, Research Funding; Xcenda: Consultancy, Speakers Bureau.
Discovery and Characterization of Chemical Inhibitors of UBC13.
Authors: Madiraju Charitha , Matsuzawa Shu-ichi , Ardecky Robert , Pass Ian , Ngo Tram , Matsuzawa Yasuko , Wimer Carina , Cuddy Michael , Diaz Paul , Ricardo Correa , Tamble Craig , Shu Chih-Wen , Pinkerton Anthony , Sergienko Eduard , Hassig Chris , Chung Thomas , Reed John C. 
Published: October 2019

Abstract
Abstract 2950
Poly-ubiquitination of signaling proteins via lysine 63 (K63)-linked chains is recognized as a critical post-translational modification involved in activation of NF-kB and stress kinases in the context of signaling by Tumor Necrosis Factor Receptors (TNFRs), Toll-like receptors (TLRs), and antigen receptors. UBC13 is a K63-specific ubiquitin conjugating enzyme that partners with TNFR-associated factors (TRAFs) to mediate K63-linked ubiquitination. Gene ablation studies have shown UBC13 is required for NF-kB signaling induced by a variety of stimuli in specific types of immune cells, making it a potential target for certain cancers, autoimmune and inflammatory diseases. UBC13 operates together with obligatory cofactors, either UEV1A in the cytosol or MMS2 in the nucleus. The nuclear function of UBC13 is evolutionarily conserved, where it plays a critical role in double strand DNA repair, making UBC13 a potential chemo- and radio-sensitizer target for oncology.
To identify chemical inhibitors of UBC13, we developed a HTS assay measuring UBC13-UEV1A enzymatic activity by TR-FRET, screening altogether ∼450,000 diverse compounds. Hit compounds were characterized using a rigorous testing funnel consisting of (a) informatics filtering against a database of > 100 HTS campaigns conducted with the same libraries, to eliminate promiscuous compounds; (b) counter-screens against E1, another cysteine-dependent enzyme (caspase-3), and against an irrelevant target formatted as a TR-FRET assay; and (c) ordering compounds from fresh powders and demonstrating reproducible concentration-dependent inhibition of UBC13. The surviving hits were then analyzed by cell-based assays for suppression of TRAF6 ubiquitination but not Mdm2-mediated ubiquitination of p53, resulting in 14 promising hits that included two chemical series. While suppressing TRAF6 ubiquitination (UBC13-dependent) in cells, these compounds did not interfere with either SUMOylation (UBC9-dependent) or NEDDylation (UBC12-dependent) of cellular proteins. UBC13 inhibitors also suppressed NF-kB activity (measured using stably integrated NF-kB-driven luciferase reporter gene) induced by PKC activators (Carma/Bcl-10/MALT pathway) and DNA damaging agent (Doxorubicin) but not by TNF-a. Investigations of the bioactivity of these UBC13 inhibitory compounds and their analogs will be described for a variety of hematolymphoid malignancies. (Supported by NIH R03-MH085677, NIH U54–005033, and by a fellowship grant from International Myeloma Foundation).

Disclosures:
No relevant conflicts of interest to declare.
Inhibition of Wnt/ß-Catenin Pathway with ICG-001 Induces Apoptosis in Multiple Myeloma.
Authors: Grigson Eileen R , Pisklakova Alexandra , Weissman Brittany , Nefedova Yulia 
Published: October 2019

Abstract
Abstract 2951
Multiple Myeloma (MM) is characterized by clonal expansion of malignant plasma cells, which preferentially reside in the bone marrow (BM). It has been previously shown that canonical Wnt signaling plays a critical role in proliferation and differentiation in MM. ICG-001 is a selective Wnt/ß-catenin signaling inhibitor that binds to the amino terminus of CREB, thus preventing ß-catenin/TCF signaling in the nucleus. Here we investigated whether the pharmacological inhibition of canonical Wnt signaling with ICG-001 could affect viability of MM cells and produce anti-myeloma effect.
C-myc and cyclin D1 are known to be downstream target genes of canonical Wnt signaling. MM cells lines treated with ICG-001 revealed down regulation of c-myc and cyclin D1 as compared to control. To investigate the varying sensitivity of cell lines to ICG-001, four cells lines were used: RPMI-8226, NCI-H929, U266, and MM1S. MM cells lines were treated with ICG-001 (1uM – 40uM) and data was collected using MTT assay or flow cytometry (Annexin binding assay). ICG-001 significantly reduced viability in all four cell lines due to induction of apoptosis, with RPMI-8226 showing the greatest sensitivity. We performed cleaved-caspase assays and demonstrated that apoptosis was caspase-3 dependent. We have also evaluated the effect of ICG-001 in BM samples from patients with MM. BM cells were treated with ICG-001 for 24 hours and the level of cleaved-caspase-3 was evaluated by flow cytometry. MM cells were defined as double positive CD138+ and Lambda/Kappa+. Our results demonstrated that MM cells were sensitive to ICG-001, while this drug was not able to induce cell death in non-tumor BM cells.
To test the effect of ICG-001 in vivo we injected SCID/Beige mice with 10 × 105 8226 cells in 100 μL phosphate-buffered saline (PBS). After 3 weeks when a tumor was measurable, the mice were split into two groups and were treated with either ICG-001 (100 mg/kg) or vehicle control (PBS) daily, for 14 days. Tumor size was consistently monitored during treatment and 3 weeks after treatment. We observed that ICG-001 produced a significant antitumor effect as compared to control group.
These data indicate targeting Wnt/ß-catenin Pathway with ICG-001 could be therapeutically beneficial to patients with MM.

Disclosures:
No relevant conflicts of interest to declare.
Identification of Multiple Subclones in Peripheral T-Cell Lymphoma, Not Otherwise Specified with Genomic Aberrations
Authors: Yoshida Noriaki , Umino Akira , Liu Fang , Arita Kotaro , Karube Kennosuke , Tsuzuki Shinobu , Ohshima Koichi , Seto Masao 
Published: October 2019

Abstract
Abstract 298
Peripheral T-cell lymphoma, not otherwise specified (PTCL, NOS) is a heterogeneous group of nodal and extra-nodal mature T-cell lymphomas that do not belong to any of the hitherto characterized T cell lymphoma subtypes. Adult T-cell leukemia/lymphoma (ATLL) is a mature T-cell neoplasm caused by human T-cell leukemia virus type 1 (HTLV-1). PTCL, NOS with genomic aberrations has been shown to resemble lymphoma-type ATLL in terms of genomic aberration patterns, histopathology and prognosis (Clin Cancer Res., 15:30–38). We have recently shown that a majority of patients with acute-type ATLL have multiple subclones that were likely produced in lymph nodes (Blood, 117:5473–5478). In this study, high-resolution oligo-array comparative genomic hybridization (CGH) was employed to determine whether PTCL, NOS with genomic aberrations also possesses multiple subclones as found in ATLL. Thirteen cases of PTCL, NOS were available for 44K high-resolution array CGH analysis. These samples analyzed in this study were shown to be monoclonal by Southern blot analysis. We obtained average log2 ratios of regions where aberrations continued over 400 probes and showed a linear average ratio of >0.1 or <-0.1. The log2 ratio imbalance was defined when a patient had a different average log2 ratio for gain or loss regions. The results showed that 11 of the 13 cases (84.6%) had a log2 ratio imbalance, suggesting that multiple subclones exist in PTCL, NOS with genomic aberrations. As previously demonstrated, the average log2 ratio of aberrant regions reflected the ratio of tumors (Blood, 117:5473–5478). In one case, as represented in Figure 1, all tumor cells had regions of loss on chromosomes 3 and 13. Fifty-four percent of tumor cells had a region of loss on chromosome 16, and 83% of tumor cells had loss on chromosome 17. It is therefore speculated that clonal evolution took place as shown in Figure 2. In an effort to examine the association between multiple subclones and prognosis, we used our previous BAC array data for 24 cases and found that the presence of multiple subclones was associated with a poor prognosis (p=0.0279) (Figure 3). Our finding concerning the presence of multiple subclones in PTCL, NOS and ATLL should assist greatly in revealing the mechanism of lymphoma development, progression and drug-resistance associated with these diseases.

Disclosures:
No relevant conflicts of interest to declare.
Tissue-Specific Homing and Different Impact in Gvhd Prevention of NK Cell Subpopulations.
Authors: Meinhardt Kathrin , Bauer Ruth , Kroeger Irena , Schneider Julia , Ganss Franziska , Dudziak Diana , Rehli Michael , Mackensen Andreas , Ullrich Evelyn 
Published: October 2019

Abstract
Abstract 3004
Clinical studies exploiting the impact of natural killer (NK) cells in allogeneic hematopoietic stem cell transplantation (HSCT) have provided promising results. It is known that NK cells are a heterogeneous population and can be divided into functionally distinct NK cell subpopulations. Murine NK cells can be separated along their expression of CD27 and CD11b and CD117 (c-kit). However, the functional relevance of distinct NK cell subsets in graft-versus-host-disease (GVHD) has not been investigated in detail so far. We have established different protocols for ex vivo isolation and expansion of murine NK cell subpopulations. These NK subsets were further analyzed in vitro and in vivo in an allogeneic murine GVHD model. Here we report on different genomic, phenotypic and functional properties of 4 NK cell subsets. Our data clearly demonstrate that CD27+ NK cells revealed the highest IFN-g production upon coculture with tumor cells and/or IL-2.
Interestingly, the CD11b+ NK cells express multiple genes of cytotoxic pathways and develop the highest cytotoxic capacity towards tumor cells. We observed up to 60% tumor lysis by CD27- CD11b+ NK cells compared to 40–45% by CD27+ CD11b+, about 25% by CD27+ CD11b- and 10% by c-kit+ CD11b- NK cells at an effector-target ratio of 5:1, respectively. Furthermore, the CD11b+ NK cell subset significantly reduced T cell proliferation induced by allogeneic dendritic cells in mixed lymphocytes reactions.
Next, we analyzed the migratory capacity and tissue-specific homing of FACS-sorted NK cell subsets by adoptive transfer of congeneic CD45.1+ and Luc+ NK cell subpopulations in autologous and allogeneic bone marrow transplantation. Of interest, FACS analysis and in vivo imaging showed that CD11b+ NK cells migrated to peripheral GVHD target organs, whereas CD27+ NK cells preferentially homed to the bone marrow.
Finally, this study addressed for the first time the role of distinct NK cell subpopulations in the development of GVHD in a fully MHC mismatched HSCT mouse model. Importantly, we identified the CD11b+ NK cell population as the NK cell subset that significantly diminished GVHD. In vivo imaging of Luc+CD11b+ NK cells revealed that this subset migrates to the colonic tissue to prevent development of GVHD colitis as shown by colonoscopy.
In summary, our comparative study outlines that only CD11b+ NK cells, migrating to the peripheral GVHD target organs and providing the most efficient cytolytic capacity directed against allogeneic dendritic cells, protect against GVHD. These new insights are highly relevant for the selection of optimal NK cell subsets in the field of cellular immunotherapy.

Disclosures:
No relevant conflicts of interest to declare.
Allogeneic Cytokine-Induced Killer Cell-Dependent Eimination of Host Dendritic Cells Leads to Less Graft-Versus-Host Disease.
Authors: Mase Shintaro , Maeba Hideaki , Fujiki Toshihiro , Kuroda Rie , Araki Raita , Koizumi Shoichi , Yachie Akihiro , Nishimura Ryosei 
Published: October 2019

Abstract
Abstract 3011
Cytokine-induced killer (CIK) cells are ex vivo-expanded T lymphocytes expressing both natural killer (NK)- and T-cell markers. We have reported that adoptive transfer of allogeneic CIK cells in a murine model caused minimal graft-versus-host disease (GVHD) with retention of antitumor activity mediated by NKG2D, which is an activating receptor expressed on NK cells. The mechanism of suppression of GVHD with CIK cells is not fully understood. Host residual dendritic cells (DCs) are most important cells in initiating GVHD reaction. Therefore, we hypothesized that alloreactive NK cells, even when infused in large numbers, do not cause GVHD by killing recipient DCs with a same mechanism as CIK cells kill tumor cells. To test this, DCs generated from rodent bone marrow cells were used for 51-Cr release cytotoxicity assays as target cells. We demonstrated that though autologous CIK cells (Balb/c) had relatively strong killing activity against mature DCs (Balb/c), allogeneic CIK cells (B6) had much more killing activity even from at a 10:1 effector -target ratio as shown in Fig 1 below. In addition, killing activity against DCs did not changed with/without adding NKG2D blocking antibody, suggesting that there were possible mechanisms for cell killing other than NKG2D/NKG2D ligand system in CIK cells. To further evaluate whether allogeneic CIK cells eliminate host DCs to reduce GVHD in vivo, lethally irradiated Balb/c recipients were given BM (B6) with CIK cells or splenocytes to compare the number and activation status of residual host-typed DCs in the spleen after bone marrow transplantation. As shown in Fig 2, on day 1 after BMT absolute number of host DCs in spleen receiving splenocytes and CIK cells were almost same, however, the numbers of DCs in the mice receiving BM alone were much less. On day 3, absolute number of host DCs in all groups decreased. In particular, the number of host DCs in the mice receiving CIK cells dramatically decreased and finally reached to the same number of host DCs in the mice receiving BM alone. On the contrary, the number of host DCs in the mice receiving splenocytes didn't decrease so much and was significantly greater than number of host DCs in the mice receiving BM alone. Considering the above, when allo-CIK cells first met to DCs, both DCs and CIK cells were expanded on day 1 after BMT, and then allo-CIK cells turned to kill host DCs. In conclusion, allogeneic CIK cells caused less GVHD due in part to elimination of host DCs.

Disclosures:
No relevant conflicts of interest to declare.
Peripheral Blood CD34+ Specific Donor Chimerism Analysis As an Early Indicator of Disease Relapse After Allogeneic Hematopoietic Stem Cell Transplant in Patients with Myelodysplastic Syndromes and Acute Myelogenous Leukemia.
Authors: Norkin Maxim , Meacham Amy , Ta Mai H , Benson Neil , Goldstein Steven , Rosenau Emma , Hsu Jack W. , Cogle Christopher R. , Hiemenz John W , Brown Randy A. , Wingard John R. 
Published: October 2019

Abstract
Abstract 3074

Background:
Disease relapse after allogeneic hematopoietic stem cell transplant (HCT) remains a major obstacle to treatment success and cure for patients with myelodysplastic syndromes (MDS) and acute myelogenous leukemia (AML). Early identification of patients at a high risk for disease relapse has significant practical implications allowing early therapeutic interventions. We evaluated a novel approach for an early identification of a post-transplant relapse in patients with MDS and AML.

Methods:
CD34+ specific donor chimerism analysis was obtained in 6 HCT recipients with CD34+ AML and MDS. Starting at day +30 post HCT CD34+ cells from peripheral blood were positively selected by immunomagnetic isolation and then underwent fluorescence-activated cell sorting. Purified CD34+ cells were subsequently evaluated for percentage of donor DNA contribution by short tandem repeat (STR) analysis. CD34+ specific donor chimerism analysis was repeated monthly until hematologic relapse or death occurred. Simultaneously, conventional donor chimerism analysis was measured in the subpopulations of peripheral blood cells.

Results:
CD34+ cells were isolated from peripheral blood with a very high purity >95% and in sufficient quantity for reliable STR analysis in all study patients. Out of 6 evaluated patients, 3 developed hematologic relapse confirmed by bone marrow evaluation. In each relapsed patient CD34+ specific chimerism dropped to <15% prior to the confirmation of hematologic relapse. However, conventional STR analysis in granulocytes, T-cells and B-cells remained >80% (Figure 1). Moreover, obvious and steady decline in CD34+ specific donor chimerism significantly preceded the time of hematologic relapse. In contrast, no decline in CD34+ chimerism was identified in 3 patients without hematologic relapse. (Figure 1).

Conclusion:
Though this is a preliminary study, the results suggest that CD34+ specific donor chimerism analysis is a useful technique for timely identification of hematologic relapse in MDS and AML patients after HCT and may provide significantly earlier relapse detection than conventional chimerism analysis.

Disclosures:
No relevant conflicts of interest to declare.
Bortezomib-Based Induction Overcomes Effect of 1q Amplification On Response in Newly-Diagnosed Myeloma and Results in Similar 2 Year Event-Free Survival
Authors: Joshua Douglas E , Hertzberg Mark S , Auguston Bradley , Spencer Andrew , Horvath Noemi , Bashford John , Campbell Lynda J , Ashurst Eleanor , Wade Jacquelyn , Copeman Michael C. , Butcher Belinda E , Prince Miles 
Published: October 2019

Abstract
Abstract 4065

Background:
Amplification of chromosome 1q occurs in 40% of myeloma and predicts lower response to induction regimens. We assessed if a PAD regimen overcomes effects of 1q amplification on overall response rate (ORR), event-free (EFS) and overall survival (OS) in newly-diagnosed, transplant-eligible myeloma.

Methods:
This phase II trial enrolled 107 patients, stratified by 1q amplification. All could receive 4× 21-day cycles of PAD induction: bortezomib 1.3mg/m2 D 1,4,8,11; doxorubicin 20mg/m2 D1,4; dexamethasone 20mg D1,2,4,5,8,9,11,12. Responders proceeded to melphalan 200mg/m2 and ASCT. ORR after PAD was primary endpoint. Secondary endpoints included ORR 3 monthspost-ASCT, and 2-year EFS and OS.

Results:
22% (20/93 evaluable cases) had amplified 1q. Median PAD cycles was4. ORRs were similar in amplified and non-amplified cases (100% vs. 87% post-PAD; 100% vs. 93% post-ASCT). After median 2 years, 2-year EFS was similar in amplified and non-amplified cases (70% vs. 75%; log-rank p=0.86), but with a trend to lower 2-year OS in amplified cases (86% vs. 94%; log-rank p=0.28). Common grade 3 or 4 adverse events included constipation (0% vs. 8% of patients), neutropenia (12% vs. 4% of patients), anaemia (4% vs. 6% of patients), back pain (4% vs. 6% of patients), neuralgia (0% vs. 6% of patients) and neuropathy (8% vs. 4%).

Conclusions:
Four cycles of PAD induced high response rates in both 1q amplified and non-amplified myeloma with acceptable toxicity. After 2 years, EFS was also similar, but with a non-significant trend to lower OS in patients with 1q amplification.

Funding:
This study is sponsored by Janssen-Cilag Pty Ltd, Australia.

Trial Registration:
ACTRN12609000296235.

Disclosures:
Joshua: Janssen-Cilag Pty Ltd: Research Funding. Hertzberg:Janssen-Cilag Pty Ltd: Research Funding. Auguston:Janssen-Cilag Pty Ltd: Research Funding. Spencer:Novartis: Honoraria; Janssen: Honoraria, Speakers Bureau; Celgene: Honoraria. Horvath:Janssen-Cilag Pty Ltd: Research Funding. Bashford:Janssen-Cilag Pty Ltd: Research Funding. Campbell:Janssen-Cilag Pty Ltd: Research Funding. Ashurst:Janssen-Cilag Pty Ltd: Employment. Wade:Janssen-Cilag Pty Ltd: Employment. Copeman:Janssen-Cilag Pty Ltd: Employment. Butcher:Janssen-Cilag Pty Ltd: Consultancy. Prince:Janssen-Cilag Pty Ltd: Research Funding.