bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Outcome of Allogeneic Stem Cell Transplantation in Chronic Lymphocytic Leukaemia: A Single Centre 10-Year-Experience
Authors: Zielinska Patrycja , Krawczyk-Kulis Malgorzata , Markiewicz Miroslaw , Dzierzak-Mietla Monika , Koclega Anna , Liwoch Robert , Kopinska Anna , Kyrcz-Krzemien Slawomira 
Published: October 2019

Abstract
Abstract 4515
Chronic lymphocytic leukemia (CLL) is an incurable disease when treated with standard chemotherapy. The only possibility to provide cure is allogeneic stem cell transplantation (allo-SCT). CLL patients aged less than 55 account for about 15% of patients and these cases allo-SCT should be taken into consideration. The indications for allo-SCT are as follows: del17p, resistance to chemoimmunotherapy, Richter’s syndrome or recurrent disease. A retrospective analysis of allo-SCT in 18 patients (10 males, 8 females) with CLL transplanted in years 2000–2010 was performed. The aim of the study was to assess of long term follow-up outcome of allo-SCT in CLL patients. The median age at diagnosis was 41ys (range: 35–51). The sibling donor was available in 16 cases (2 pts were mismatched), unrelated donors were in 2 cases (1 mismatched). Most of the pts (16 out of 18) were MRD positive when allotransplanted. Median lymphocytosis preceeding allo-SCT was 5.9G/l. Peripheral blood was the source of stem cells in 9 cases (50%), and bone marrow in the remaining 9 cases, 2 pts were transplanted with stem cells from bone marrow and peripheral blood. 4 pts (22%) underwent the allograft procedure twice or more. Reduced intensity conditioning with alemtuzumab was performed in 9 pts (50%), myeloablative regimen in 4 cases and RIC with rituximab in one case.The median number of CD34+cellsx10^6/kg was 4.1 (range: 0.86–9.64). All but one patient engrafted (this pt was transplanted again successfully in one year time). Acute graft-versus host disease (GvHD) was noted in 46% of pts (only in 2 pts grade IV). Extensive GvHD was observed only in 2 pts. Donor lymphocyte infusion (DLI) was performed in 8 pts (44%). With a median follow-up of 73 months (range: 9–89) for surviving patients, the five-year Kaplan-Meier of overall survival (OS) and progression free survival (PFS) was 55,5% and 34%, respectively. At five years, the cumulative probability of non-relapse mortality was 15%. Allogeneic stem cell transplantation remains the effective treatment in CLL for selected group of patients.

Disclosures:
No relevant conflicts of interest to declare.
Effect Of Plerixafor Mobilization On Immune Reconstitution Post High Dose Chemotherapy and Autologous Stem Cell Transplantation
Authors: Solh Melhem M. , Smith Tori , Khaled Yasser , Copik Alicja 
Published: October 2019

Abstract

Introduction
Plerixafor is a reversible CXCR4 antagonist that has been approved by the food and drug administration for autologous hematopoietic stem cell mobilization in patients with multiple myeloma and non-Hodgkin’s lymphoma. Patients mobilized with Plerixafor were shown to have a higher proportion of primitive stem cells (CD34+/CD133+/CD38-), CD4+ T cells and Natural killer cells (CD3-/CD16+/CD56+) in the graft composition when compared to patients mobilized with chemotherapy plus G-CSF alone .We investigated the effect of Plerixafor on immune reconstitution at thirty and sixty days post autologous stem cell transplantation.

Methods
Patients eligible for autologous stem cell transplantation were enrolled on a single arm prospective immune reconstitution trial. A complete blood count, differential and lymphocyte flow cytometry panel (T cell, NK cell and B cell markers) was checked on Days 30 and 60 post autologous transplantation. Stem cell mobilization was carried per our institutional standards. All patients received subcutaneous G-CSF at a dose of 10 µg per kilogram body weight for four consecutive days. On Day4, patient with a peripheral CD34 count of ≤20/µl received plerixafor 0.24mg/kg. Collection was started on day 5 and continued till collection goal was reached or patient failed to get the minimal cell dose after 4 consecutive days of aphaeresis.

Results
49 patients were enrolled during the period from September 2010 till May 2012. Median age at time of transplantation was 54 years (range 21; 72 years). 35 patients had multiple myeloma and 14 had non-Hodgkin’s lymphoma. 16 patients received GCSF alone (group A) and 33 had plerixafor plus GCSF (group B) for mobilization. All patients achieved the minimum target of CD34 collection. The mean number of collection days was 1.9 and 1.4 days (p=0.05) with a total collection dose of 7.76 and 7.61 CD34 x106/Kg for groups A and B respectively. The percentage proportion of CD34 in the aphaeresis product was 0.73% and 0.75% (p=0.9) for group A and B. Total infused CD34 dose was similar in both groups (4.88 and 4.56 CD34x106/kg) with time to engraftment of 11.68 vs 11.69 days for neutrophils and 20.62 vs 21.39 for platelets in groups A and B respectively. There was no difference between day 30 absolute lymphocyte count (1.09 vs 1.44 x103/mm3 p=0.18); Absolute NK cell (0.31 vs 0.35 x103/µl; p=0.51); absolute T cell count (0.71 vs 0.96 x103/µl p=0.33) and absolute neutrophil count (2.98 vs 2.63 x103/mm3 p=0.37). The cell count recovery was also not significantly different when analyzed per disease (myeloma or non-Hodgkin’s lymphoma) and at day 60.

Conclusion
Our study shows that patient mobilized with plerixafor and G-CSF have similar immune reconstitution at 30 and 60 days post autologous transplantation compared to patients mobilized with G-CSF alone.

Disclosures:
No relevant conflicts of interest to declare.
Effect Of Allogeneic Blood Transfusion On Peripheral Blood Τregs and Cytokine Levels Of Patients That Underwent Joint Replacement Surgery
Authors: Spadidea Panagiota , Rodi Maria , Chatziantoniou Alexandros , Panagiotopoulos Elias , Mouzaki Athanasia 
Published: October 2019

Abstract

Purpose/Objective
Clinical and experimental studies have established that allogeneic blood transfusion (ABT) can cause immunosuppression. To identify immune parameters that contribute to this effect, we determined the effect of ABT on peripheral blood (PB) cytokine profiles and Treg numbers and function in a cohort of patients with no underlying pathologies.

Materials and Methods
Heparinized PB samples were collected from 46 patients (7M/39F, 28-88 yo) that underwent joint replacement surgery. The samples were collected immediately before surgery (BS), and after surgery (AS) on days 0, 7, 1 month, and 3 months to 1 year. Thirty six patients received ABT and 10 did not. PBMC were isolated, and the numbers and % of CD4+CD25+Foxp3+ Tregs and CD4+CD25high/+CD127low/- Tregs were determined by FACS. Tregs and T effectors (Teff) were isolated from patients on days 0-7 and Treg functional assays were performed by culturing Tregs with PHA-stimulated Teff at different ratios for 72h with CFSE, and analyzed by FACS for proliferation. Cytokine levels were determined in plasma by a cytometric bead array assay for IL-2, IL-4, IL-5, IL-6, IL-10, TNF-á, IFN-ã, and ELISA for TNF-á, TNF-RI(p55/p60) and II(p75/p80), TGF-â1 and â2.

Results
Both, CD4+CD25+Foxp3+ and CD4+CD25high/+CD127low/- Treg populations increased significantly on d0 AS and decreased on d7 AS to below BS levels, to achieve homeostasis >3 months in transfused patients. In contrast, Treg levels remained the same between BS and AS in non-transfused patients.
Functional assays showed that Tregs were functional post-ABT and could suppress Teff proliferation efficiently. In culture, isolated Tregs secreted TGF-â1.
All cytokines and TNF-RI and II plasma levels increased on d0 AS (IL-6, TNF-RI and II significantly), in transfused patients immediately AS, and decreased by d7 AS. In contrast, TGF-â1 levels decreased on d0 AS and increased by d7. No differences in cytokine or receptor levels were observed in non-transfused patients AS.

Conclusion
ABT induces the immediate production of plasma cytokines, especially pro-inflammatory IL-6 and TNF-RI and II that decreases by d7. In parallel, ABT induces the proliferation of Tregs. These Tregs are functional, and secrete TGF-â1 that reaches BS plasma levels by d7. The Tregs seem to be Th3 inducible Tregs. Homeostasis is reached by 3 months post-ABT.

Disclosures:
No relevant conflicts of interest to declare.
OSU-T315, An Integrin-Linked Kinase (ILK) Inhibitor, Induces Apoptosis By Targeting B Cell Receptor and CD49d Mediated AKT/ERK Activation In Chronic Lymphocytic Leukemia Cells
Authors: Liu Ta-Ming , Ling YongHua , Phelps Mitch A. , Woyach Jennifer A. , Hertlein Erin K , Towns William H. , MacMurray Jessica , Salunke Santosh B. , Chen Ching-Shih , Byrd John C. , Johnson Amy J. 
Published: October 2019

Abstract
Aberrant regulation of endogenous survival pathways plays a major role in pathogenesis of chronic lymphocytic leukemia (CLL). Signaling via conjugation of surface receptors within the tumor environmental niche is believed to activate survival and proliferation pathway in CLL. PI3K/AKT appears to be the pivotal axis to support CLL growth. Current studies identify Integrin-linked kinase (ILK) as a Phosphoinositide-dependent kinase-2 (PDK2), by which it mediates AKT activation and concomitant with tumorigenesis in many solid tumors. As a result of our search for novel therapeutic targets, we first report the potent inhibitor, OSU-T315, modeled from the scaffold that docks into AKT-binding site of ILK in therapeutic of hematological malignancy. OSU-T315 triggers apoptosis in CLL by targeting both intrinsic and extrinsic AKT and ERK activation (Figure 1A). OSU-T315 induces cytotoxicity to both CLL-derived cell lines Mec-1 and OSU-CLL, as well as in primary CLL cells, while illustrating alleviated toxicity in normal B and T cells (Figure 1B, C). In contrast to the highly successful BTK and PI3 kinase inhibitors which inhibit BCR signaling proximally, OSU-T315 specifically inhibits down-stream AKT and ERK signaling cascades without affecting proximal BCR or focal-adhesion signaling. In particular, external stimuli through BCR or CD49d, by which triggers survival signal and protects CLL from ankoisis via AKT/ERK activation, is potently inhibited by OSU-T315. Downstream anti-apoptotic molecule Mcl-1 is suppressed upon OSU-T315 treatment, which is later accompanied by increased Caspase 3/7 activity. Further study indicates that OSU-T315 mediated cell death is Caspase dependent. In vivo efficacy of OSU-T315 is supported by a therapeutic study in the TCL1 mouse model of CLL, from which demonstrates improvement in total counts of white blood cells and reduced splenomegaly, thereby contributing to enhanced overall survival following OSU-T315 oral treatment regimen (Figure 2). Together, our findings suggest the innovative dual-targeted approaches by suppressing both AKT and ERK pathway in CLL therapy through inhibiting ILK, and provide an alternative agent for potential therapeutic development in CLL.

Disclosures:
No relevant conflicts of interest to declare.
CD49d Is The Strongest Flow Cytometry-Based Predictor Of Overall Survival In Chronic Lymphocytic Leukemia
Authors: Bulian Pietro , Shanafelt Tait D , Fegan Chris , Zucchetto Antonella , Cro Lilla , Nuckel Holger , Baldini Luca , Kurtova Antonina , Ferrajoli Alessandra , Burger Jan A. , Gaidano Gianluca , Del Poeta Giovanni , Pepper Chris , Rossi Davide , Gattei Valter 
Published: October 2019

Abstract

Purpose
Although CD49d is an unfavorable prognostic marker in chronic lymphocytic leukemia (CLL), definitive validation evidence is lacking. A worldwide multi-center analysis was performed using published and unpublished CLL series to evaluate the impact of CD49d as overall survival (OS) and treatment free survival (TFS) predictor.

Patients and Methods
A training/validation strategy was chosen to find the optimal CD49d cut-off. The hazard ratio (HR) for death and treatment imposed by CD49d was estimated by pooled analysis of 2,972 CLL, and Cox analysis stratified by center and stage was used to adjust for confounding variables. The importance of CD49d over other flow cytometry-based prognosticators (CD38, ZAP-70) was ranked by recursive partitioning.

Results
Patients with ≥30% of neoplastic cells expressing CD49d were considered CD49d+. The decrease in OS at 5 and 10-years among CD49d+ cases was 7% and 23% (decrease in TFS 26% and 25% respectively). The pooled HR of CD49d for OS was 2.5 (2.3 for TFS) in univariate analysis. This HR remained significant and of similar magnitude (HR=2.0) in a Cox model adjusted for clinical and biological prognosticators. Hierarchical trees including all cases, or restricted to early stage or patients ≤65 years, always selected CD49d as the most important flow-cytometry-based biomarker, with negligible additional prognostic information added by CD38 or ZAP-70. Consistently, by bivariate analysis, CD49d reliably identified patients' subsets with poorer outcome independent of CD38 and ZAP-70.

Conclusions
In this analysis of ∼3000 patients, CD49d emerged as the strongest flow cytometry-based predictor of OS and TFS in CLL.

Disclosures:
Shanafelt: Genentech: Research Funding; Glaxo-Smith-Kline: Research Funding; Cephalon: Research Funding; Hospira: Research Funding; Celgene: Research Funding; Polyphenon E International: Research Funding. Burger:Pharmacyclics: Research Funding.
First US Phase I Clinical Trial Of Globin Gene Transfer For The Treatment Of Beta-Thalassemia Major
Authors: Boulad Farid , Riviere Isabelle , Wang Xiuyan , Bartido Shirley , Prockop Susan E. , Barone Rita , Moi Paolo , Maggio Aurelio , Sadelain Michel 
Published: October 2019

Abstract

To date, the only curative therapeutic approach for beta-thalassemia major has been allogeneic stem cell transplantation (SCT) for patients with HLA-matched siblings. For the majority of patients who do not have a matched sibling, allogeneic SCT is associated with major risks of morbidity and mortality. The stable transfer of a functional globin gene into the patient’s own hematopoietic progenitor cells (HPCs) yields a perfectly matched graft that does not require immunosuppression to engraft. We previously demonstrated successful globin gene therapy in murine thalassemia models, using a lentiviral vector that encodes the human ß-globin promoter and arrayed regulatory elements uniquely combined to achieve high level and erythroid-specific globin expression. In vivo in thalassemic mice, the vector termed TNS9.3.55, increased hemoglobin levels by an average 4-6 g/dL per vector copy. We obtained in 2012 the first US Food and Drug Administration (FDA) approval to proceed to a clinical study in adult subjects with beta-thalassemia major (NCT01639690). We have to date enrolled 5 patients and recently treated the first three, administering the transduced HPCs after non-myeloablative conditioning. Engraftment data are available for the first two patients. Patient 3 was recently infused with CD34+ cells and is at this time too early to evaluate.
Patient 1 is a 23 year old female with a ß039 – IVS1,110 mutation. Patient 2 is an 18 year old female with a ß039 – IVS1,6 mutation. Both patients underwent mobilization of peripheral blood stem cells (PBSCs) with filgrastim and mobilized 25 x 10^6 and 9.9 x 10^6 CD34 cells/Kg respectively. CD34+ PBSCs were transduced with the lentiviral vector TNS9.3.55 encoding the normal human beta-globin gene. The average vector copy number (VCN) in bulk CD34+ cells for these two patients was respectively 0.39 and 0.21 copies per cell. Both patients underwent non-myeloablative cytoreduction with busulfan administered at 2 mg/Kg/dose Q12H x 4 doses (total 8 mg/Kg), followed by reinfusion of 11.8 x 10^6 and 8.4 x 10^6 CD34+ cells/Kg, respectively. Both patients tolerated cytoreduction well and recovered their blood counts. While they continue to be transfusion dependent, both patients show a gradual rise in vector copy number in peripheral blood white blood cells and neutrophils, steadily increasing by 1-2% every month, reaching an average VCN of 5-7% 3-6 months after transplantation.
In summary, patients with thalassemia major underwent safe and effective mobilization followed by excellent transduction of mobilized CD34+ cells. The transplant non-myeloablative conditioning was well tolerated, and followed by rapid engraftment and gradual rise in VCN. Continued clinical and molecular monitoring is on-going and will be presented.

Disclosures:
No relevant conflicts of interest to declare.
Characterizing the Global Transcriptional Effects of TEL-AML1-Fusion in Childhood Acute Lymphoblastic Leukemia
Authors: Teppo Susanna , Teittinen Kaisa J , Grönroos Toni , Viiri Keijo , Kaikkonen Minna , Heinäniemi Merja , Lohi Olli 
Published: October 2019

Abstract
Acute lymphoblastic leukemia (ALL) is the most common childhood cancer affecting thousands of children worldwide each year. Most patients are cured but only after long chemotherapy treatment and still 15 % of them relapse. Therefore, better understanding of the molecular biology behind the disease is fundamental. TEL-AML1 fusion - comprised of two transcription factors important in hematopoiesis regulation - is the most frequent genetic variation found in 25 % of pediatric ALL-patients. The translocation occurs during pregnancy and is suggested to serve as a “first hit” enabling secondary changes that lead to cancer formation. Despite its frequency, genetic targets of this aberrant transcription factor are unclear, and its function in leukemia initiation and progression remains elusive.
Here we aim at solving the genome-wide effects of TEL-AML1 on transcription using pre-B-leukemic cell models. After a time-series induction of wild-type or DNA-binding compromised mutated TEL-AML1, or a constitutive expression of shRNA silencing the fusion, the cells were studied using global nuclear run-on sequencing (GRO-seq). GRO-seq technology can map RNA polymerase II location and identify actively transcribed coding and non-coding genes and their directionality. Thus, GRO-seq provides a snapshot of global active transcription and was used here to identify bona fide targets of the TEL-AML1-fusion protein.
In addition to transcriptional changes in known genes, we identified a notable number of novel transcripts including antisense-, lincRNA-, and microRNAs. We also characterized genomic locations where the fusion functions as a direct regulator by taking advantage of non-coding RNA transcripts known as enhancer RNAs. Furthermore, we performed transcription factor motif enrichment analysis on these active regulatory eRNA regions. The results uncover early transcriptional changes induced by TEL-AML1-fusion and may reveal critical steps in leukemia initiation and development. The findings also highlight the importance of non-coding RNAs in the pathogenesis of pediatric leukemia.

Disclosures
No relevant conflicts of interest to declare.
In Vitro Preactivation of PBMCs with PM-mb21-41BBL Particles Stimulates in Vivo Expansion of NK Cells Under Low IL-2 in NSG Mice
Authors: Oyer Jeremiah L , Pandey Veethika , Igarashi Robert Y , Colosimo Dominic A , Solh Melhem M. , Altomare Deborah A , Copik Alicja J 
Published: October 2019

Abstract
NK cell immunotherapy as a cancer treatment shows promise, but methods for consistent expansion of NK cells from a small fraction (~5%) of peripheral blood mononuclear cells (PBMCs) to therapeutically effective amounts are not widely accessible. Strategies that promote in vivo expansion rely on high-dose IL-2 in combination with lymphodepletion and suffer from severe treatment related toxicities as well as limited expansion. On the other hand, in vitro methods are available for robust NK cell expansion, but rely on prolonged in vitro co-culture with feeder cells that is costly and complex and requires high dose IL-2 (25,000-35,000 U) for persistence after injection into mice. An optimal NK cell expansion method would rely on off the shelf reagents, would not require long-term complex cultures, and would induce a rapid, sustained in vivo expansion of NK cells from unsorted PBMCs under low concentration of IL-2. We have developed a method to prepare particles from K562 cells engineered to express specific cytokines, and such particles stimulate NK cell expansion. Particles prepared using K562 cells expressing IL-21 and 41BBL (PM21 particles) cause highly specific expansion of cytotoxic NK cells from unsorted PBMCs that achieves 95% NK cells in ~14 days and ~10,000 fold expansion of NK in about 21 days. NK cell expansion is consistent using different preparations of PM-particles or leukocyte sources, and the PM-particles retain expansion efficacy with storage. The PM-mb21-41BBL particles were also used to stimulate in vivo NK cell expansion in NSG mice under ultralow IL-2 (1,000 units per injection, 3 injections per week, per mouse). Unsorted human PBMCs, either shortly pre-activated in culture for two days or not pre-activated, were injected (2 x 106 PBMCs per mice by i.p.) and human CD56+CD3- NK cells and other relevant hCD45+ lymphocytes were monitored in the peripheral blood over time and in tissues and fluids at the time of euthanization. In vivo NK cell expansion was observed in mice injected with PBMCs that were pre-activated with PM21 particles. The extent of NK cell expansion observed in the peripheral blood was in levels that would be relevant for clinical cancer treatment (>50,000 NK cells/mL of mouse blood 14 days post injection). In vivo NK cell expansion was further confirmed with analogous experiments using PBMCs that were stained with CellTrace Violet to monitor proliferation. Notably, NK cells were also found in spleen, bone marrow, lung, liver and brain. These results taken together provide proof of principle that PM-particle technology is effective for direct in vivo NK cell expansion from unsorted PBMCs under low dose IL-2.

Disclosures
No relevant conflicts of interest to declare.
Utilization Trend and in-Hospital Complications of Autologous Hematopoietic Stem Cell Transplantation in Multiple Myeloma in the United States: 13 Years Perspective
Authors: Savani Bipin N. , Mukherjee Akash , Savani Ghanshyambhai T , Ankit Chothani 
Published: October 2019

Abstract
Background: We aimed to explore utilization, complication and in-hospital mortality related to Autologous Hematopoietic Stem Cell Transplant (ASCT) in Multiple myeloma (MM) from 1998 to 2011 in United States.
Methods: We used Nationwide Inpatient Sample (NIS) database to isolate multiple myeloma cases undergoing ASCT. Study included adults age >18 years. Patient and hospital demographics were extracted from NIS. Peri-procedural complications were calculated using ICD-9-CM codes. Hierarchical multivariate regression models were formulated to assess predictors of in-hospital mortality and complication.
Results: Total of 6,957 (weighted-34,837) were available for analysis. Majority of patients were male (57%), white (57%) who presented to large bed size (78%),teaching (96%) hospital. Utilization of ASCT for MM increased from 5% in 1998 to 21.6% in 2011 (p<0.001). Despite temporal increase in mean age and comorbidites, mortality reduced (2.9 vs. 0.7, p<0.01) significantly from 1998 to 2011. In-hospital complications were common after ASCT with stomatitis (44.7%) being the most common followed by anemia requiring transfusion (28.5%), febrile neutropenia (16.5%) and bacteremia (15.0%). Mechanical ventilation and acute respiratory failure are the strongest predictors of in hospital mortality post ASCT followed by acute kidney injury,total Body irradiation (TBI) and bacteremia. Higher Charlson comorbidity index, female sex and TBI were associated with higher complication rates post ASCT.
Conclusions: Utilization of ASCT for MM patients has increased significantly since 1998 and even though complications are very common in patients undergoing ASCT (77.4%), in-hospital mortality is rare (<2%) and has decreased significantly over the years secondary to decreased use of pre transplant TBI and better transplant care.
Figure 1 Figure 1.
Figure 2 Figure 2.
Figure 3 Figure 3.

Disclosures
No relevant conflicts of interest to declare.