bluuebunny/crossref_metadata_2025_split huggingface dataset:
RUNX1C-Mediated Reprogramming Of Human Bone Marrow Stromal Cells into Early Blood Progenitors
Authors: Yin Weihong , Porada Christopher D , Walker Stephen , Bishop Colin , Almeida-Porada Graca 
Published: October 2019

Abstract
Somatic cell reprogramming to the hematopoietic lineage, either through a pluripotent state or directly, opens the possibility of production of a ready source of autologous hematopoietic stem cells (HSC) that can be used to treat/cure a wide variety of blood disorders. While it has previously been shown that dermal fibroblasts (HFF) can be directly reprogrammed to the hematopoietic lineage, the efficiency was relatively low and the resultant hematopoietic cells lacked multilineage differentiative potential. Stro1(+) isolated stromal progenitors (SIPs) can easily be isolated from the bone marrow (BM) and expanded ex-vivo to obtain clinically significant numbers of cells. In similarity to HSC, SIPs are derived from the mesoderm, and are intimately linked with HSC specification during ontogeny. As such, they are likely to be epigenetically closer to HSC than HFF, and therefore good candidates for reprogramming into hematopoietic cells. To verify the uniqueness of SIPs for reprogramming, we transduced SIPs and HFF with OCT4 and/or RUNX1C, a master transcription factor (TF) that triggers the developmental onset of definitive hematopoiesis, in the following combinations: 1) OCT4 alone; 2) RUNX1C alone; or 3) OCT4+RUNX1C. We then performed a timeline of gene/cell surface marker expression (using microarray, qRT-PCR, and flow cytometry) from day 3-16 post-transduction. Visual inspection of the cultures showed that, while reprogrammed colonies began to appear in SIPs cultures at day 9, no colonies were seen during this time period in HFF cultures. Flow cytometry and molecular analyses of colonies obtained from OCT4+RUNX1C combination demonstrated that expression of CD41, the earliest marker of commitment to the hematopoietic lineage, commenced within only 3-4 days and peaked at day 5-6, by which time ∼20% of SIPs expressed this marker. This peak in CD41 expression coincided with commencement of expression of CD34 and CD45, and maximal induction of several hematopoiesis-specific TFs and phenotypic markers such as PU.1, HOXB4, GATA2, MIXL, WNT3, KDR, CDX4, which occurred at 1-3 logs higher levels in SIPs than HFF. Further studies demonstrated that the chromatin remodeling function of OCT4 could be replaced with the histone methyltransferase inhibitor Bix-01294, with the combination of RUNX1C and Bix-01294 inducing levels of CD34 and CD41 expression by day 5 that were similar to those achieved with RUNX1C plus OCT4. The present studies thus take several important steps towards making the promise of producing autologous hematopoietic cells for transplantation via direct reprogramming a reality.

Disclosures:
No relevant conflicts of interest to declare.
Phase II Randomized Study Of Lenalidomide Or Lenalidomide and Rituximab As Maintenance Therapy Following Standard Chemotherapy For Patients With Intermediate-High/High Risk Diffuse Large B-Cell Lymphoma (DLBCL)
Authors: Reddy Nishitha M. , Morgan David S. , Park Steven I. , Greer John P , Richards Kristy L. 
Published: October 2019

Background
DLBCL patients (pts) with an intermediate/high to high risk international prognostic index (IPI) are at an increased risk of disease relapse rate in the first year after completion of standard therapy with R-CHOP. Lenalidomide (len), an immunomodulatory drug, has activity in relapsed diffuse large B cell Lymphoma. Len enhances the natural-killer cell mediated antibody-dependant cellular cytotoxicity of rituximab in lymphoma cell lines and inhibits angiogenesis as well as alters cytokine production. Lenalidomide received FDA approval based on its clinical activity in relapsed mantle cell lymphoma.

Methods
Intermediate-high/high risk IPI pts with DLBCL were randomized to len (arm A) alone or len and rituximab (arm B). The primary endpoint of the study was to assess the one year disease-free survival. We expected that a 25% difference of relapse will have clinical significance when compared with current standard therapy. Pts in arm A received len at a dose of 25 mg daily for 21 days of 28 days. Patients on arm B received len at a dose of 20mg daily for 21 days of 28 days along with rituximab on day 8 of even cycles. Treatment on both arms was continued for one year. Treatment was discontinued for disease progression. Len dose adjustments are incorporated in the protocol.

Results
Forty three pts, 21 arm A/ 22 arm B, 21 female/22 male, with a median age of 59 yrs were enrolled. The median IPI was 4 for pts over the age of 60 and the median aa-IPI was 3. Three pts received XRT to areas of bulky disease. At a median follow up of 27 months, the 2 yr DFS and OS was 88%. For patients in arm A and arm B the 2 yr DFS was 90% vs. 86% (figure 1) and the 2 yr OS was 95.2% vs. 81% (figure 2), respectively (P=NS). Two pts discontinued treatment due to adverse events. Grade 3-4 toxicities include neutropenia (28%), fatigue (16%), diarrhea (6%), DVT (3%), rash (3%), febrile neutropenia (3%). Related grade 1-2 toxicities include hypothyroidism (15%) and rash (54%).

Conclusions
Len as maintenance therapy demonstrates clinical activity following standard chemotherapy and improves DFS and OS in DLBCL patients with high risk prognostic features as compared with historical controls. This trial is registered with NCI- #NCT00765245

Disclosures:
Reddy: Celgene: Research Funding. Off Label Use: lenalidomide in diffuse large B cell lymphoma. Park:TEVA: Research Funding; Seattle Genetics, Inc.: Research Funding.
Multiple Myeloma Cells Express IL-17A Creating An Autocrine Loop: An Attractive Therapeutic Target
Authors: Prabhala Rao H. , Fulcinitti Mariateresa , Pelluru Dheeraj , Prabhala Harsha K. , Rashid Naim , Sperling Adam , Nanjappa Puru , Lee Saem , Negroiu Andreea , Lazo-Kallanian Suzan , Munshi Aditya , Richardson Paul G. , Daley John , Anderson Kenneth C. , Munshi Nikhil C. 
Published: October 2019

Abstract
We have previously demonstrated that Th17 cells, which produce IL-17A, are significantly elevated in peripheral blood and bone marrow (BM) of patients with Multiple Myeloma (MM) and IL-17A promotes MM cell growth and survival, both in vitro and in vivo via IL-17A receptor. We have recently evaluated and observed that anti-IL-17A monoclonal antibody (mAb) significantly inhibited MM cell growth in vitro, while IL-17A induced proliferation of MM cells compared to control. We have also observed significant down-regulation of IL-6 production by anti-IL-17A mAb in MM-BMSC co-culture. Importantly, the administration of anti-IL-17A mAb weekly for 4 weeks in the SCIDhu model of human myeloma, where MM cells grow within the human microenvironment in mice led to a significant inhibition of tumor growth compared to the control mice. This remarkable activity of anti-IL17 mAb raised the question of whether the myeloma cells themselves are a possible source of IL-17. In this study, we used transcriptome sequencing (RNA-Seq) data to evaluate the expression of IL-17A in primary CD138+ myeloma cells (N=17) compared to normal plasma cells (NPC) (N=5). Whereas none of the NPCs expressed IL-17A, it was significantly over-expressed in majority of MM cells. In addition, these data also showed that the expression of other IL-17 family members (IL-7B, C, D, E & F) and Th17-associated pro-inflammatory cytokines (IL-21, IL-22 & IL-23) were not significantly elevated in primary myeloma cells compared to normal donor plasma cells. We further validated these observations by IL-17 immunoblot showing IL17 expression in all MM cell lines and 10 out of 14 primary patient MM cells; confirmed IL-17 expression in MM cells by quantitative RT-PCR, and flow cytometry and by immuno-histochemistry and confocal microscopy. We observed that IL-17 knock down by IL-17-specific siRNA inhibited MM cell growth as well as their ability to induce IL-6 production in co-cultures with BMSC. Finally, expression profile data from 172 uniformly treated patients showed that patients with lower IL-17A expression had superior overall survival compared to those with higher expression. These data confirms that MM cells express IL-17 and targeting it with a mAb will abrogate the autocrine loop making it an attractive therapeutic target.

Disclosures:
No relevant conflicts of interest to declare.
Development Of a Serum Biomarker Panel That Predicts Imminent Risk Of Multiple Myeloma (MM) Progression From Premalignancy
Authors: Kate Sasser A , Casneuf Tineke , Amaratunga Dhammika , Tryputsen Volha , Kumar Shaji K , Rajkumar Vincent , Hall Brett 
Published: October 2019

Abstract
Premalignant states are known to precede malignancy in many different cancers, including multiple myeloma (MM). Each year, 1% of Monoclonal Gammopathy of Undetermined Significance (MGUS) patients and 10% of Smoldering Multiple Myeloma (SMM) patients transition to symptomatic MM, which remains an incurable disease. By identifying patients at imminent risk (within 2 years) of progression to MM, treatment could begin early and potentially prevent full blown MM and associated morbidity and mortality. Multiple risk-factor models are currently utilized to identify patients at high-risk of developing MM. Commonly cited risk factors include percentage of bone marrow plasma cells, free light chain ratio, serum levels of M-protein, IgG subtype and levels of circulating plasma cells. These tests have individual and combined merit but there remains no consensus for measuring imminent risk of MM progression. To determine whether serum proteomics could help predict progression or provide information on relevant pathways involved in progression to MM, we evaluated 200 MGUS and SMM (187 non-IgM) patient samples on the Myriad DiscoveryMAP(r) platform (250+ serum proteins), supplemented by demographic and clinical information. Of the 187 non-IgM patients, 100 progressed to MM within 2 years (high-risk), and 87 did not progress to MM within 5 years (low-risk). We performed different subset analyses to screen for proteins that were informative and predictive of disease progression from MGUS/SMM to MM. First, a univariate Wilcoxon rank sum test was performed. The top 10 proteins that were found to be differential between high-risk/low-risk for all MGUS and SMM patients were IgM, Progesterone, MCP-2, Kallikrein-5, FGF4, EPO, MCP-4, MIP-1β, MCP-1, and TF. Interestingly, when analyzing MGUS and SMM patients separately, additional proteins that were important in distinguishing high-risk/low-risk patients included IgM, EN-RAGE, BAFF, and several others previously shown to be relevant in MM, as well as a handful of novel proteins with unidentified roles in MM. Second, to determine whether specific combinations of proteins could be utilized together to define high/low-risk, a classification tree was assembled. The combination of 8 features (IgM, Progesterone, Aldose Reductase, E-Selectin, Fas Ligand, Hemoglobin, Age and Creatine kinase-MB) appropriately classified 86% of the 187 patients into high/low-risk. Future analyses will include examining this high/low risk classifier in contrast to current risk factor models, and application to two additional sets of similar patient samples that serve as an independent validation set.

Disclosures:
No relevant conflicts of interest to declare.
C950T and C1181G Osteoprotegerin Gene Polymorphisms In Myeloma Bone Disease
Authors: Sonmez Mehmet , Kazaz Nazli , Yucel Burcu , Topbas Murat , Ucar Fahri 
Published: October 2019

Abstract
Bone disease is one of the hallmarks of multiple myeloma (MM). The role of osteoprotegerin (OPG) in the RANK/RANKL/OPG signaling system is well defined in the myeloma bone disease. Polymorphisms of the TNFRSF11B gene encoding OPG have been studied in various bone diseases. However, relationship between the levels of OPG and development of bone lesions regardless of RANKL is yet unknown. In this study, the effects of OPG gene polymorphism on the development of bone lesions in MM were investigated.
52 MM patients, admitted to Karadeniz Technical University, Medical Faculty, Department of Internal Medicine, Division of Hematology between March 2011 and March 2012, were included into the study. Pathologic fracture and/or destructive bone lesions were confirmed in 36 MM patients using X-ray, computed tomography (CT), or magnetic resonance imaging (MRI). No bone lesions were noted in 16 patients. The control group consisted of 20 age and gender matched volunteers.
Exon 1 of OPG gene was sequenced using DNA samples obtained from peripheral blood samples of 52 MM patients and 20 healthy control subjects. OPG gene C950T and C1181G polymorphisms were identified in 45 (33 with bone lesions) MM patients and in 16 control subjects. Polymorphic allele 1181G was higher in MM patients with bone lesions compared to MM patients without bone lesions and control group (χ2=8.629, p=0.013). Other polymorphic allele 950T was also overrepresented (73%; p=0.042) in MM patients with bone lesions compared to MM patients without bone lesions and the control group. The most frequent genotype observed in patients with myeloma bone lesions was 950 TT homozygous variant (56%, p=0.08) followed by 1181 CC homozygous wild type with 50% frequency (p=0.09). The frequency of 950 CT heterozygous genotype was higher in MM patients without bone lesions (50%, p=0.604). 1181 GG genotype showed higher frequency in MM patients with bone lesions compared to patients without bone lesions and control group (χ2=3.853, p=0.145). Of the combined haplotypes, 950 TT/1181 GG haplotype frequency is higher in patients with myeloma bone lesions (36%) compared to MM patients without bone lesions (13%) and controls (15%) (χ2=4.77, p=0.09). The frequency of TT/GG combined haplotype carriers in bone disease patients is higher than of those who do not have either polymorphic homozygous alleles (CT/CC and CC/CC haplotypes) (χ2=6.057, p=0.048). Additionally, 950 CC/1181CC wild type haplotype frequency was reasonably low in patients with myeloma bone lesions (8.3%, p=0.240).
This is the first study searching for the relationship between OPG gene variants C950T (promoter), C1181G (exon 1) and myeloma bone disease. It was concluded that the presence of polymorphic 1181 G/950 T alleles and 950 TT/1181 GG genotypes may play a role in the development of bone disease.

Disclosures:
No relevant conflicts of interest to declare.
Combined Bendamustine, Prednisone and Bortezomib (BPV) In Patients With Relapsed Or Refractory Multiple Myeloma and Light Chain Induced Renal Failure
Authors: Pönisch Wolfram , Moll Barbara , Niederwieser Dietger 
Published: October 2019

Abstract

Introduction
Serious renal failure represents a main complication of Multiple Myeloma (MM). An estimated 25% to 50% of patients are affected during the course of their disease. These patients are at increased risk for infections and have a significantly worse prognosis. Small phase I/II studies suggest that treatment with chemotherapy and/or new substances results in recovery of renal function in more than 25%. The window of opportunity for reversal of renal impairment is rather small making an immediate and highly active treatment strategy mandatory. Bortezomib and bendamustine have turned out to be quickly acting and effective drugs in the treatment of MM.

Methods
Between March 2005 and March 2013, 36 patients (median age 64; range 32-81 years) with relapsed/refractory MM and light chain induced renal failure (creatinine clearance <60ml/min) were treated with bendamustine 60mg/qm on day 1 and 2, bortezomib 1.3mg/qm on day 1, 4, 8 and 11, and prednisone 100mg on day 1, 2, 4, 8 and 11. Cycles were repeated every 21 days. Patients were divided into two groups: group A (n=20) consisted of patients with moderate or severe renal dysfunction (eGFR 15-59ml/min) and group B (n=16) of patients with renal failure/dialysis (eGFR <15ml/min).

Results
The median number of the BPV-treatment was 2 (1-7) cycles. 24 patients (67%) responded after at least one cycle of chemotherapy with 3 CR, 3 nCR, 6 VGPR, and 12 PR. Six patients had MR, 2 patients stable disease and 4 patients had a progress. With a median follow up of 22 months of the surviving patients, median PFS and OS for patients with moderate or severe renal dysfunction (group A) were 10 months and 25 months, respectively. Outcome for these patients was significantly better compared to patients with renal failure/dialysis (group B) with a median PFS and OS of 3 months and 7 months, respectively (p<0.02).
Eleven patients showed a CRrenal, 5 patients a PRrenal and 15 patients a MRrenal. Median time to first renal response and best renal response were 21 days and 42 days, respectively. The most common severe side effect was grade 3-4 thrombocytopenia in 81% of the patients. Grade 3-4 neutropenia was observed in 50% of the patients. Moderate to severe infections were seen in 13 patients.

Summary
These results indicate that the combination of bortezomib, bendamustine and prednisone is effective and well tolerated in patients with relapsed/refractory MM and light chain induced renal failure.

Disclosures:
No relevant conflicts of interest to declare.
JAK1/2 Inhibition Suppresses T Cell Proliferation and Inflammatory Cytokine Production In An Allogeneic Setting and Ameliorates Graft-Versus-Host Disease (GvHD) In a Murine GvHD Model
Authors: Spoerl Silvia , Chen Sophia , Bscheider Michael , Mathew Nimitha , Schmickl Martina , Mueller Tony , Schmitt-Graeff Annette , Peschel Christian , Duyster Justus , Poeck Hendrik , Zeiser Robert , von Bubnoff Nikolas 
Published: October 2019

Abstract
Graft-versus-host-disease (GvHD) constitutes a severe complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). In GvHD, tissue damage is mediated by pro-inflammatory cytokines. Cytokine responses are mediated by activated Janus kinases (JAKs). We hypothesized that JAK1/2 inhibition might reduce T effector cell responses and inflammatory cytokine production in an allogeneic system, thereby ameliorating GvHD. We established an allogeneic (major mismatch) cell culture system using naive BALB/c CD4+ CD62Lhigh T cells that were co-cultured with C57BL/6 (B6) bone marrow derived dendritic cells (DC). JAK 1/2 signaling was specifically blocked using the small molecule inhibitor Ruxolitinib (INCB018424). By using a 3H Thymidine proliferation assay, we found that Ruxolitinib was able to inhibit the proliferation of allogeneic CD4+ effector T cells. In our intracellular cytokine staining experiments Ruxolitinib was able to impair the differentiation of naïve T cells into IFN-gamma and IL17A-producing T effector cells. Both cytokines – IFN-gamma and IL-17A – have been linked to aggravated courses of GvHD severity.
In vivo administration of Ruxolitinib signficantly reduced lethal GvHD after allo-HSCT in a murine major mismatch model. BALB/c recipient mice were irradiated and received C57BL/6 (B6) T cell depleted bone marrow along with CD4+ and CD8+ T cells. Animals that were treated with an oral gavage of Ruxolitinib twice daily displayed significantly lower serum levels of IFN-gamma, TNF-alpha, IL-10 and IL-12p70, lower histological and clinical GvHD scores and improved overall survival compared to the control group.
By using luciferase-positive (luc+) CD4+ and CD8+ T cells, we were able to visualize and quantify T cell migration after GvHD induction. The recipient mice received luc+ T cells along with conventional T cell depleted bone marrow. We were able to detect the luc+ T cell signal with a bioluminescence imaging system. At day 9 after allogeneic bone marrow transplantation, migration of donor T cells to GvHD target sites was significantly reduced in Ruxolitinib treated mice compared to control mice.
In summary, our data demonstrate that suppression of inflammatory cytokine responses by JAK1/2 inhibitors modulates T effector cell responses in an allogeneic setting and improves survival in a murine major mismatch GvHD model. These observations suggest that JAK1/2 inhibition might be used for the treatment of GvHD following allo-HSCT.

Disclosures:
No relevant conflicts of interest to declare.
Influence Of Race and Ethnicity On The Collection Of G-CSF Mobilized Peripheral Blood Stem Cells From Unrelated Donors, a Center For International Blood and Marrow Transplant Research Analysis
Authors: Hsu Jack W. , Wingard John R. , Logan Brent R. , Chitphakdithai Pintip , Shaw Bronwen E. , Goldstein Steven C. , O’Donnell Paul V. , Confer Dennis L. 
Published: October 2019

Abstract

Peripheral blood stem cell (PBSC) collection is increasingly used in allogeneic stem cell transplantation. However, a small percentage of healthy donors have a poor mobilization response to G-CSF. Very little information exists on the effect of donor race or ethnicity on PBSC mobilization. We analyzed 10776 unrelated donors from the National Marrow Donor Program (NMDP) who underwent G-CSF mobilized PBSC collection from 2006–2012. We investigated the effect of self-reported donor race/ethnicity on collection efficiency, defined as number of CD34+ cells/L (of donor blood processed), number of mononuclear cells (MNC)/L and CD34+ cells/MNC collected on the first day of apheresis. Categorical variables were analyzed by the Chi-square test and the Kruskal-Wallis test was used for continuous variables. A linear regression model was used to compare the various race/ethnic groups while controlling for potential confounding factors (such as age, BMI, gender, and year of apheresis). The result of our analysis is shown in Table 1. Univariate analysis revealed statistically significant differences in CD34+ cells/L, MNC/L and CD34+/MNC in all races analyzed. In general, African Americans (AA) had the highest collection efficiency while Caucasians had the lowest. Other races/ethnicities had collection efficiencies between the two groups. On multivariate analysis, statistically significant differences in CD34+ cell/L were seen in Hispanics, AA and Asian/Pacific Islanders (API), primarily in the obese (Hispanic, AA, API) and overweight (AA, API) donors. In the API group the differences in collection efficiency were predominately seen in males. No differences were seen between Caucasians and Native Americans. This study reveals significant racial/ethnic differences in the efficiency of collection of CD34+ cells in unrelated donors. Although these differences do not appear to interfere with the ability to collect adequate numbers of PBSC, it is currently unknown why they exist. This is an area for continued research.

Disclosures:
No relevant conflicts of interest to declare.