bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lenalidomide and Low Dose Dexamethasone in Previously Treated Multiple Myeloma Patients with Impaired Renal Function: Efficacy Analysis of PrE1003, a Precog Study
Authors: Mikhael Joseph , Manola Judi , Lonial Sagar , Weiss Brendan M , Oettel Kurt , Hayman Suzanne R. , Kanate Abraham S. , Rajkumar S. Vincent 
Published: November 2019

Abstract
Introduction: Lenalidomide (Len) and dexamethasone (dex) has proven to be a highly effective treatment for multiple myeloma (MM) and serves as the basis of other combinations in relapsed disease. However, nearly 1/3 of myeloma patients have renal insufficiency, and the optimal use of Len in this population is not well known. We undertook this study in 3 cohorts of pts: Group A had creatinine clearance (CrCl) of 30-60 ml/min, Group B with CrCl < 30 ml/min, and Group C, with CrCl < 30 ml/min and requiring dialysis. The Phase I component of the trial has been previously reported (ASCO 2014) demonstrating that full dose Len (25mg daily 21/28 days) can be given to all patients despite renal insufficiency, even when on dialysis. We now present data on response to treatment.
Methods: In the Phase 2 component accrual was slow and the trial was terminated. However, prior to termination a modified exploratory analysis plan was established, including all 34 patients treated at the recommended phase 2 dose. This plan would declare the treatment worthy of further study if 18 or more of the 34 patients responded to treatment. This design had 84% power to distinguish a response rate of 60% from a null rate of 40%, using a 1-sided test with 10% type I error.
Results: The Phase 2 efficacy cohort accrued 34 patients, consisting of 20 patients from Group A, 9 patients from Group B, and 5 patients from Group C. Response of PR or greater was seen in 17 patients, resulting in a 50% overall response rate (CI 37-63%). Treatment was well tolerated with expected adverse events; the most common adverse events were anemia, diarrhea, and fatigue. Eleven episodes of Grade 3 and one Grade 4 pneumonia were reported for 10 patients across all cohorts and dose levels; 3 were considered possibly related to treatment. Seventeen patients have remained on treatment for more than 12 cycles. In the exploratory analyses, median PFS was 7.5 months (95% CI, 3.6 - 19.7 months) and median OS was 19.7 months (95% CI, 10.4 - 42.5 months).
Conclusions: Len Dex can be safely administered to patients with impaired renal function at full dose of 25mg daily 21/28 days, and is associated with a 50% response rate. Further investigation into other combinations with Len Dex in patients with renal impairment should be considered.
Table Table.

Disclosures
Mikhael: Onyx: Research Funding; Sanofi: Research Funding; Abbvie: Research Funding; Celgene: Research Funding. Lonial:Merck: Consultancy; Novartis: Consultancy; Onyx: Consultancy; BMS: Consultancy; Onyx: Consultancy; Celgene: Consultancy; BMS: Consultancy; Novartis: Consultancy; Celgene: Consultancy; Janssen: Consultancy; Millenium: Consultancy; Janssen: Consultancy. Weiss:Novartis: Consultancy.
Evaluation of Various Strategies to Generate NPM1mut-Specific T Cell Clones
Authors: Ruecker-Braun Elke , Heidenreich Falk , Link Cornelia S , Schmiedgen Maria , Wehner Rebekka , Kühn Denise , Heidrich Katharina , Alakel Nael , Schmitz Marc , Thiede Christian , Ehninger Gerhard , Bornhäuser Martin , Schetelig Johannes 
Published: November 2019

Abstract
Mutated nucleophosmin (NPM1) was identified as a promising leukemia-specific antigen for cytotoxic T lymphocytes (CTL). NPM1 is a multifunctional nucleocytoplasmic shuttling phosphoprotein. In AML patients with normal cytogenetics NPM1 mutations are the most frequent molecular genetic abnormalities, accounting for up to 60% of the patients. The peptide (AIQDLCLAV) derived from the mutated NPM1 (NPM1mut) has been described to elicit a CTL response restricted to HLA-A*02:01. We observed that NPM1mut multimer+ T cells were very rare in peripheral blood. The limitation of the multimer technology is the absence of a positive control; nevertheless it is an attractive tool to generate antigen positive T cell clones.
The goal was to compare strategies for the generation of NPM1mut multimer+ T cell clones systematically. For this purpose we analyzed blood samples from two patients with AML after transplantation and six different healthy donors.
We explored different strategies to isolate HLA-A*02:01 restricted NPM1mut multimer+ T single cells. The first strategy was to isolate multimer+ T cells directly from the blood without any supplements by single cell sorting. The second strategy was to sort multimer+ T cells which were previously CD8+ enriched supplementing the media either with or without IL-21. Published by Yongqing et al.IL-21 enhances the generation of human antigen-specific CD8+ T cells. A further strategy was to previously enrich CD14+ cells for the generation of autologous monocyte-derived dendritic cells (MoDCs). The co-cultivation of MoDCs loaded with the NPM1mut peptide and CD8+ cells were performed either with or without IL-21, as well. We expanded the last strategy by a second round of NPM1mut-specific stimulation.
So far it was not possible to generate NPM1mut-specific T cell clones based on the advanced strategies and consistently there is no data published on NPM1mut multimer+ T cell clones. This fact raises the question why NPM1mut specific clones display such low frequencies. We want to point out that although we varied the strategies and we used eight different donors the isolation of NPM1mut-specific T cells restricted to HLA-A*02:01 apparently is challenging. Greater efforts, e.g. a larger number of donors or the use of immunological checkpoint inhibitors during cell culture are needed.

Disclosures
Thiede: AgenDix: Employment, Other: Ownership. Schetelig:Sanofi: Honoraria.
Imaging Human Immune Cell Infiltration in a Xenograft Graft-Versus-Host Disease Model
Authors: Elssen Catharina Van , Rashidian Mohammad , Vrbanac Vladimir , Freund Christian , Cragnolini Juanjo , Ingram Jessica , Tager Andrew , Ploegh Hidde 
Published: November 2019

Abstract
Graft versus host disease (GvHD) is one of the major complications of hematopoetic stem cell transplantation (HSCT), and is characterized by massive alloreactive T cell infiltration into target organs and tissues. Currently, non-invasive tools to diagnose and follow up disease are lacking. Here we show non-invasive imaging of infiltrating immunocytes in a human xenograft model for chronic GvHD, using a single domain camelid-derived antibody fragment (VHH) specific for human Class II MHC products present on myeloid cells, B cells and activated T cells. NSG (NOD-scid gamma) mice reconstituted with human fetal thymus, bone marrow and liver (BLT mice) spontaneously develop a GvHD-like condition, characterized by alopecia, blepharitis and target organ infiltration by activated human T cells. In diseased animals, we show an increased PET signal in the liver, attributable to infiltration of activated Class II MHC positive T cells. Non-invasive imaging of specific immune infiltration and activation could thus be of importance for diagnosis and treatment evaluation of GvHD and holds promise for other diseases characterized by inflammation.

Disclosures
No relevant conflicts of interest to declare.
Dendritic Cell Production from Allogeneic Donor Cd34+ Stem Cells and Mononuclear Cells; Cancer Vaccine
Authors: Unal Ali , Birekul Ayse , Unal Mehmet Cagri , Karakus Esen , Köker Yavuz , Ozkul Yusuf , Comu Faruk Metin 
Published: November 2019

Abstract
Aim: Active immunotherapy provides better recognition of tumor-related antigens by immune system of patient, enhanced immune system and elimination of malignant cells. This modality employs therapeutic potential of donor specific and tumor specific immune responses. Active immunotherapy targets immunosuppressive and tolerogenic mechanisms suppressed by tumor cells. T lymphocytes and antigen-presenting cells (dendritic cells) are two cell lineages that play crucial role in the battle of organism against cancer. Close similarity between cancer cells and normal cell structure is the most important reason of escape from defense cells, namely T lymphocytes. Stimulation and enhancement of T lymphocytes against cancer cells comprise principal part of therapy.
Material and Method: To generate allogeneic dendritic cells, leukemic stem cells were isolated from bone marrow samples from patients with acute leukemia. Lysate was prepared from leukemic stem cells identified by flow cytometer. Stem cells and mononuclear cells (1x10>6/kg) obtained from sibling donors by apheresis were separated to produce Dendritic Cells. For Dendritic Cell transformation, GM-CSF and IL-4 were added to media where leukemic stem cell lysate from patient and mononuclear cells from sibling donor were treated. From samples taken from the culture medium, after 48, 72 and 96 hours, dendritic cell surface markers (CD 80, CD 83 and CD 86) was assessed by flow cytometry. CD3, CD14, CD19, CD56, CD66b-negative, and HLA-DR, CD86 positive are indicative of immature DC. HLA-DR, CD 80 and CD83 positive are indicative of mature DC.
Results: Mononuclear cells were detected by 27 % among allogeneic hematopoietic cell groups harvested by apheresis. After culture under GMP conditions, mononuclear cell rate was found to be 24% on hours 96 and 120. It was seen that 88% of mononuclear cells transformed to mature dendritic cells after 96 hours culture.
Conclusion: In cancer patients, minimal residual disease can be eliminated by active tumor vaccine after reducing tumor burden by standard methods. Tumor vaccine obtained from allogeneic sibling donor can be used in lieu of autologous tumor vaccine and it is thought to be more effective. Allogeneic dendritic cells produced at 37¡C in CO2 media under GMP conditions can be used in tumor immunotherapy. More effective method would have been used by employing dendritic cells against cancer stem cells rather than cancer cells itself.
Figure Mature dendritic cells (CD83+) Figure. Mature dendritic cells (CD83+)

Disclosures
No relevant conflicts of interest to declare.
The Role of MLL1 and MLL2 in MLL Fusion Oncoprotein-Initiated Leukemia
Authors: Ernst Patricia , Anastassiadis Konstantinos , Kranz Andrea , Stewart A. Francis , Arndt Kathrin , Waskow Claudia , Yokoyama Akihiko , Jones Kenneth , Neff Tobias , Chen Yufei 
Published: November 2019

Abstract
The MLL1 histone methyltransferase gene at 11q23 undergoes many distinct chromosomal translocations to yield poor prognosis leukemia. In these MLL-rearranged (MLL-r) leukemias, the remaining wild-type allele is typically expressed, and the fusion oncoprotein lacks the C terminal histone methyltransferase domain. The role of the wild-type, endogenous MLL1 allele in MLL-fusion protein mediated leukemogenesis is not fully understood, with some evidence suggesting an important functional role. On the other hand, the wild-type allele is lost in some patients and in the MLL-r ML2 cell line, suggesting that the wild-type allele is not absolutely required. Here we rigorously address the role of endogenous MLL1 in MLL-r leukemia. Using precise and independent animal models, we show that MLL1 is dispensable for MLL-AF9- or MLL-AF6-mediated acute myelogenous leukemia (AML). In parallel experiments, deletion of Menin, encoding a chromatin targeting cofactor for MLL-fusion proteins, has severe effects on the proliferation and survival of MLL-r leukemia, as previously published. Potential redundancy between MLL family methyltransferases was addressed by co-deleting the closest paralog, Mll2. Surprisingly, deletion of Mll2 by itself had a significant impact on survival of MLL-AF9 leukemia cells, and co-deletion of Mll1 and Mll2 more effectively killed these cells, suggesting a redundant or synthetic lethal relationship between the two genes. A comparison of gene expression and histone modification changes in Mll1, Mll2 and Mll1;Mll2 double knockout leukemia cells showed that 1) global defects in histone H3, lysine 4 methyation can be exclusively attributed to MLL2, 2) MLL1- versus MLL2-dependent genes are largely distinct, and 3) Mll1;Mll2 double knockout leukemia cells exhibit deficiencies in NFκB and integrin ß3 pathways, but exhibit little evidence of deregulation of either MLL1- or MLL-fusion protein target genes. These findings reshape our thinking about the role of endogenous MLL family proteins in MLL-fusion protein leukemogenesis. The data presented here, in conjunction with the minimal effect of Mll2 deletion on hematopoietic stem and progenitor cells make MLL2 an attractive candidate for targeted therapy in MLL-r leukemia.

Disclosures
Ernst: Amgen: Other: stocks. Yokoyama:Sumitomo Dainippon Pharma: Research Funding. Neff:Bristol Myers Squibb: Other: Travel; Janssen: Other: Travel; Epizyme: Patents & Royalties: patent filed.
Comparison of the Terumo BCT MNC and Cmnc Protocols for Peripheral Blood Stem Cell Collections
Authors: Westbrook Lindsey , Roig Joseph , Bagamasbad Neil , Dilag Reynold , Nasser Melissa , Sauthoff Sheri , Berg Mary 
Published: November 2019

Abstract
Background: Terumo BCT recently offered a new method of peripheral blood stem cell (PBSC) collection using the Spectra Optia¨, an apheresis instrument. The new protocol includes a continuous mononuclear cell collection (CMNC) as opposed to an older version, the mononuclear cell collection (MNC), which involves an additional step where product pools through a cell separation insert. Our institution has used both methods and the purpose of this study was to compare the CMNC to the MNC protocol, including run times and PBSC product characteristics.
Methods: A retrospective review and comparison of parameters from 120 collection procedures using the MNC insert and 60 collection procedures using the CMNC insert was done using the t-test. Data from patients/donors (including 20 allogeneic donors) as well as procedure details including run time, flow cytometry marker for stem cells (CD34)-positive (CD34+) throughput, CD34+ collection efficiency (CE%), platelet loss (plt loss/total blood volume [TBV]), and collection product characteristics were included in the analysis.
Results: The MNC donor group included 12 allogeneic donors, which is comparable to the 8 allogeneic donors in the CMNC group. Donor weight and patient weight was not significantly different between the two groups. Pre-procedure laboratory values (WBC, percentage of lymphocytes [lymph%], percentage of monocytes [MNC%], and platelet count) were also similar between the two groups.
Run time was found to be significantly shorter using the CMNC protocol compared to the MNC protocol. Product volume was also significantly lower in the CMNC group compared to the MNC group. Although the volume was lower, the CMNC product had significantly higher white blood cell count (WBC), MNC%, and lymph% when compared to the MNC product. The CD34+ throughput was significantly higher in the CMNC group than the MNC group. The CD34+ CE% was found to be slightly increased in the CMNC group, though not significantly. The platelet loss was nearly identical in both protocols when normalized for total blood volume. Product hematocrit (HCT%) was significantly higher using the CMNC protocol; however, the red blood cell volume never exceeded 20 mL due to the lower product volume with the CMNC protocol. The numerical results are summarized in the Table.
Conclusion: The CMNC protocol collects a smaller volume of a purer product when compared to the MNC protocol with comparable platelet and red blood cell loss. Staff members who perform apheresis procedures are pleased by the shorter run time.
Table Table.

Disclosures
Roig: Terumo BCT: Employment.
Hematopoietic Recovery after in-Vivo T-Cell Depleted Allogeneic Stem Cell Transplant- Effects of ABO Incompatibility, Rhesus Incompatibility and Acute Gvhd
Authors: Gergis Usama , Al-Mulla Naima Ali , Javier Segovia , Hafez Rania , Phillips Adrienne A. , Mayer Sebastian , Shore Tsiporah B. , Hsu Jing-ming , Slotky Ronit , Vasovic Ljiljana V. , Cushing Melissa M. , van Besien Koen 
Published: November 2019

Abstract
Background:
Transfusion needs after allogeneic transplantation are closely associated with morbidity, cost of transplantation and may also be associated with long-term outcome. In an effort to better predict determinants of recovery after reduced intensity conditioning, we analyzed 214 consecutive patients who received allogeneic hematopoietic stem cell transplantation (HSCT) at our institution from January 2012 to December 2013.
Patients and Methods:
We excluded patients who experienced early (in the first 4 months) relapse (n=28), non-relapse mortality (n=28) or who experienced prolonged hospital stay before day 100 for transplant related complications (n=51). One hundred and seven patients had uneventful recovery until day 100 and met the inclusion criteria. The majority of patients received Fludarabine and melphalan conditioning (87%). Approximately one third each of the patients had HLA-identical related, HLA-matched unrelated donors or underwent haplo-cord transplantation. Recipients of HLA-identical related or unrelated donor transplant received alemtuzumab and post-transplant tacrolimus. Haplo-cord recipients received thymoglobulin and post-transplant tacrolimus and mycophenolate. Patients and transplant characteristics are summarized in table 1. T-tests were used for comparison between groups.
Results: In our analysis of 107 patients who underwent T cell depleted allogeneic transplantation and fared well at day 100, the determinants of prolonged anemia were ABO incompatibility (p=0.006), rhesus incompatibility (p=0.01) and acute graft versus host disease (aGVHD) (p=0.02). Whereas prolonged thrombocytopenia was only associated with the development of any grade aGVHD (p=0.04). At day 100, the hematopoietic recovery of haplo-cord grafts is similar to matched related and unrelated grafts.
Conclusion:
ABO incompatibility, rhesus incompatibility and occurrence of acute graft vs host disease are the major determinants of red blood cell recovery. Occurrence of acute graft vs host disease is a major determinant of platelet recovery. Of interest, the use of umbilical cord blood grafts combined with haplo-identical grafts is associated with recovery of red blood cells and platelets that is similar to that of adult donor grafts.

Disclosures
No relevant conflicts of interest to declare.
Comparison of Reduced Intensity Conditioning (RIC) Regimen and Myeloablative Conditioning (MAC) Regimen in Allogeneic Hematopoietic Stem Cell Transplantation (Allo-HSCT) for Adult Patients with Acute Leukemia
Authors: Gurman Gunhan , Aydemir Guloksuz Gozde , Atilla Erden , Ataca Atilla Pinar , Civriz Bozdag Sinem , Kurt Yuksel Meltem , Toprak Selami Kocak , Topcuoglu Pervin , Arslan Onder , Ozcan Muhit , Demirer Taner , Ilhan Osman , Akan Hamdi , Beksac Meral 
Published: November 2019

Abstract
Introduction:The development of RIC regimens enabled HSCT also for elderly patients who had contraindications for intensive conditioning regimens. The aim of this study is to evaluate the effects of conditioning regimens on outcomes of alllo-HSCT for adult patients with acute leukemia.
Method:In this study, we retrospectively analysed 490 acute leukemia patients who underwent allo-HSCT between June 1988 to June 2015 at Ankara University School of Medicine Department of Hematology. We compared the outcomes of MAC vs RIC conditioning by Fisher's test and chi-square test. Kaplan-meier curves were used for survival. The factors that effect on survival were evaluated by univariate analysis-log-rank test and multivariate analysis-cox regression.
Results: In our study, the median age of participants was 32 with M/F ratio of 285 (57.9%) / 207 (42.1%). The patients were diagnosed as acute myeloid leukemia (AML) in 344 patients (69.9%), acute lymphoblastic leukemia (ALL) in 140 patients (28.5%) and others in 8 patients (1.6%). Pre-transplant active disease detected in 105 (22.7%) patients. Patients received grafts from related donors in 397 (80.7%) transplants. The most common stem cell source was peripheral blood in 378 patients (76.8%) followed by bone marrow in 99 patients (20.1%). MAC was used in 224 (90.4%) recipients whereas RIC was used in 47 (9.6%). The age, unrelated transplants, number of AML patients and intermediate-high risk patients were significantly higher in RIC group (44 vs 31, p<0.001; 37% vs 18%, p=0.002; 81.4% vs 43.7%, p<0.001). Both 1-year OS and 1-year RFS rate were lower in the patients treated with RIC (65.6% vs 34.9%; 44.4% vs 16.7%). Among the patients treated with MAC, 1 year-OS was higher in patients who developed chronic GVHD (78.6% vs 51.1%, p<0.05). The higher OS was related with patients who were transplanted in remission, received grafts from related donors as well as having acute and chronic graft vs host disease.
Conclusion: Lower OS and RFS were observed in the RIC conditioned patients compared to MAC in our study. In the treatment of young patients who do not have comorbidities for intense conditioning regimens, show good performance status and are high risk; the initial use of standard myeloablative conditioning regimens is feasible. However, the lower number of RIC transplants and the variations between groups in terms of patient and donor characteristics might effect the results of this study. Therefore, conducting further prospective studies with larger cohorts would be beneficial to enhance our knowledge.

Disclosures
No relevant conflicts of interest to declare.