bluuebunny/crossref_metadata_2025_split huggingface dataset:
Haploidentical Peripheral Blood Stem Cell Transplantation with Post-Transplantation Cyclophosphamide in Primary Refractory Acute Myeloid Leukemia
Authors: Jaiswal Sarita Rani , Chatterjee Sumita , Chakrabarti Aditi , Bhargava Sneh , Kunal Ray , Chakrabarti Suparno , O'Donnell Paul V. 
Published: October 2019

Abstract
In a pilot study, 75 patients with Primary Refractory (PRef) AML without matched family donors were offered post-transplantation cyclophosphamide (PTCY) based haploidentical peripheral blood stem cell (PBSC) transplantation. Twenty-seven patients (36%) opted for haploidentical transplantation with or without further chemotherapy. There was no difference in the patient or disease characteristics amongst patients undergoing transplantation or not. The conditioning regimen comprised of FluCyMel (n=5), FluBuMel (n=17) and FluTreoTBI (n=5). MMF was tapered between days 14 and 21 posttransplant in the absence of GVHD and cyclosporine A was tapered between days 60 and 90. The progression free survival at a median follow-up of 25 months was 36.6% and 0% in the transplant and the non-transplant group (p=0.0001). Prompt engraftment was noted at a median of 14 days irrespective of disease status or conditioning regimens. Cumulative incidences of acute graft-versus-host disease (GVHD) and chronic GVHD were 26.6% and 8% respectively. The overall incidence of infections remained low, with CMV reactivation and invasive aspergillosis occurring in 9 and 2 patients respectively. CMV disease was diagnosed in 2 patients. Non-relapse mortality at 1 year was 16.7%. The incidence of disease progression was 54%. Factors positively impacting progression free survival were < 15% marrow blasts at transplant and a Natural Killer Cell Ligand Mismatch (NKLMM) donor. NKLMM, Haplotype or B scores had no impact on CMV infection or GVHD. However, Bx Haplotype was associated with lower NRM (5%, 95%CI-1-9) compared to 48.6% (95%CI 28.2-69.0) in AA Haplotype (p=0.01). Disease status did not impact the overall survival (p=0.11) in the HSCT cohort. In fact, NKLMM donors with B haplotype had the greatest impact on overall survival in both the HSCT cohort (71.4%, 95%CI 54.3-88.5%) compared to 20% (95%CI 8.8-31.2, p=0.01) in those without the same. Our data suggests PTCY based Haploidentical PBSC transplantation is feasible in patients with PRef AML and donor NKLMM might improve progression free survival, provided the conditioning protocol and the post-grafting therapy offer the optimum platform for alloreactivity of NK cells.

Disclosures
No relevant conflicts of interest to declare.
A Novel Recombinant Eklf-GATA1 Fusion Protein Reduces Erythrocytes Sickling in Human Erythroid Culture Model
Authors: Zhu Jianqiong , Chin Kyung , Aerbajinai, Wulin , Li Hongzhen , Kumkhaek Chutima , Hsieh Matthew , Rodgers Griffin P 
Published: October 2019

Abstract
Current gene therapy approaches for treatment of hemoglobinopathies involve viral transduction of hematopoietic stem cells with "antisickling" globin genes. Hemoglobin A2 (HbA2, α2δ2), expressed at a low level due to the lack of Eklf binding motif in its promoter region, is fully functional and could be a valid substitute for hemoglobin A in β-thalassemia, as well as an "anti-sickling" agent in sickle cell disease. We have previously demonstrated that two Eklf-GATA1 fusion proteins, which were recruited to the GATA1 binding motif at the δ-globin promoter, can significantly activate δ-globin expression in K562 cells and hematopoietic stem CD34+ cells. Here, we report that enforced expression of Eklf-GATA1 fusion protein in sickle trait CD34+ cells significantly increased δ-globin expression, as determined by quantitative PCR and HPLC. Upon deoxygenation, the percentage of sickling cells was lower in Eklf-GATA1-transduced red blood cells as compared with mock-transduced cells. By a series of flow cytometry analyses, which includes BRDU incorporation assay, CD71/GPA, and thiazole orange staining, we found that erythroid cells proliferation, differentiation and enucleation were not affected by Eklf-GATA1 expression. To assess the potential off-target effects of our fusion protein constructs, we analyzed differentially expressed genes (and proteins) in vector-only and Eklf-GATA-1 transduced-CD34+ cells by microarray and proteomic changes by liquid chromatography-mass spectrometry (LC-MS). We found that over-expression of Eklf-GATA1 resulted in a less than 2-fold change in the gene expression profile related to bone marrow hematopoiesis, and there were no significant changes that were detected in proteomic profiling. These results indicate that these fusion constructs could be a valuable genetic therapeutic tool for hemoglobinopathies, and warrant further preclinical study and evaluation.

Disclosures
No relevant conflicts of interest to declare.
Higher BTK-Dependent Cell Proliferation in Unmutated Chronic Lymphocytic Leukemia Confers Increased Sensitivity to Ibrutinib
Authors: Guo Ailin , Lu Pin , Galanina Natalie , Nabhan Chadi , Smith Sonali M. , Coleman Morton , Wang Yue Lynn 
Published: October 2019

Abstract
In chronic lymphocytic leukemia (CLL), mutation status of the immunoglobulin heavy chain variable region gene (IGHV) has prognostic significance, with the unmutated phenotype (UM-CLL) having worse outcomes than mutated CLL (M-CLL) following chemotherapy or chemoimmunotherapy. However, in the era of BCR-targeted therapies, the adverse prognostic impact of unmutated IGHV seems to be diminishing, and there are clinical datasets showing unexpected improved responses in UM-CLL. We investigated the biological differences of BTK activity between these subgroups and further compared the impact of ibrutinib on molecular and cellular behaviors. Immunoblotting analysis revealed that phosphorylated active BTK. is significantly higher in UM-CLL. Moreover, UM-CLL, compared to M-CLL, displayed a much higher proliferative capacity that was associated with greater sensitivity to ibrutinib. In addition, BTK depletion with siRNA led to a more prominent reduction in the proliferation of UM-CLL, suggesting that elevated BTK activity is responsible for increased cell proliferation. Further, cell signaling activity by multiple measurements was consistently higher in UM-CLL accompanied by a higher sensitivity to ibrutinib. These studies link UM-CLL to elevated BCR signaling, higher BTK-dependent cell proliferation and increased sensitivity to ibrutinib. The prognostic significance of IGHV mutation should be reevaluated in the era of new therapies targeting BCR signaling.

Disclosures
Nabhan: Celgene Corporation: Honoraria, Research Funding. Smith:Celgene: Consultancy; Pharmacyclics: Consultancy.
Validation of the Nextseq 500 and Development of a High-Throughput NGS Pipeline for Identifying Clinically-Relevant Gene Variants in Multiple Myeloma Specimens
Authors: Zielinska Agnieszka K. , Leigh Kenton , Gomez Horacio , Van Laar Ryan K 
Published: October 2019

Abstract
As molecular profiling technologies have evolved, our understanding of multiple myeloma heterogeneity and the relative effectiveness of treatment options have increased dramatically. Most recently, next generation sequencing (NGS) studies have provided a new degree of molecular resolution into this disease, however it remains a challenge to translate these methodologies and insights from research tools to widely-available clinical assays which can be performed as part of routine patient care.
MyPRS® is a clinically and scientifically validated high-throughput gene-expression (Affymetrix) based assay available in all 50 US states. The CLIA and CAP-accredited laboratory workflow is able to isolate sufficient RNA from small amounts of fresh bone marrow aspirate, up to 72 hours post collection[1]. In order to expand the content of the MyPRS assay to include DNA-sequencing based variant, copy number and mutation detection, we have validated highly-scalable methods for isolating RNA and DNA separately and in parallel from individual patient specimens of varying quality and quantity.
Yet another challenge of translational NGS profiling is performing complex laboratory procedures, developed primarily for research use only, with the requisite reproducibility and accuracy required for submission to regulatory agencies and ultimately for clinical use. Coupled with the protocols we developed for isolation of both DNA and RNA from small amounts of patient bone marrow aspirate, we performed an investigation of the Illumina NextSeq 500 and "on-site" BaseSpace data processing server. With the multiplexing and parallel processing capability of this platform we estimate being able to sequence, align and variant-call up to 150 myeloma-relevant genes at 1000x coverage per run.
Results from our MM cell-line-, normal human- and NIST reference-DNA whole-exome-profiling studies show extremely high levels of technical reproducibility and agreement with results generated from 3rd party laboratories. In addition, we describe associations between the (RNA-expression based) prognostic, molecular subtyping and virtual karyotyping currently included in the MyPRS assay, with results from DNA-based exome profiling, performed on matched specimens. We believe findings underscore the need for comprehensive DNA and RNA based molecular profiling in order to make the most informed patient management decisions.
1. van Laar R, Flinchum R, Brown N, Ramsey J, Riccitelli S, Heuck C, Barlogie B, Shaughnessy Jr J: Translating a gene expression signature for multiple myeloma prognosis into a robust high-throughput assay for clinical use. BMC Medical Genomics 2014, 7 (1):25.

Disclosures
Zielinska: Signal Genetics: Employment. Leigh:Signal Genetics: Employment. Gomez:Signal Genetics: Employment. Van Laar:Signal Genetics: Employment.
Plasmocytosis and Immunoparesis but Not Neutrophil to Lymphocyte Ratio Can Predict Time to Treatment in Smoldering Myeloma
Authors: Romano Alessandra , Consoli Marzia L , Parisi Marina , Cavalli Maide , Parrinello Nunziatina L , Conticello Concetta , Di Raimondo Francesco 
Published: October 2019

Abstract
Objectives: We recently identified the ratio between absolute neutrophils count and absolute lymphocyte count, NLR ≥2, as a predictor of progression free survival (PFS) and overall survival (OS) in patients younger than 65 years with symptomatic Multiple Myeloma (MM). We retrospectively examined the NLR in a cohort of 140 smoldering Myeloma (sMM) defined accordingly to the updated IMWG 2014 guidelines accessed our Center between June 2006 and June 2014.
Methods: NLR was calculated using data obtained from the complete blood count (CBC) at diagnosis and subsequently correlated with time to treatment (TTT) for symptomatic MM. All patients underwent bone marrow evaluation to estimate plasma cells infiltration (BMPC), Magnetic Resonance Imaging (MRI) to detect bone lesions, serum free-lite chain evaluation (sFLC) starting from January 2012 when they became evaluable in our center.
Results: The mean NLR was 2.0 ± 0.1, lower than the value previously found for MM (2.7 ± 0.2, p= 0.005). Higher NLR was independent of BMPC amount, cytogenetics and sFLC. Using NLR ≥2 as predictive biomarker we could not predict TTT.
In univariate analysis only BMPC ≥ 30% (p<0.0001), immunoparesis (suppression of one or more uninvolved immunoglobulins, p=0.017) and presence of at least one bone lesion at MRI (p<0.0001) could predict TTT. In multivariate analysis, these three parameters were independent (p<0.0001). Since new guidelines consider BMPC≥ 60% and positive MRI as myeloma-defining events, we proposed a simplified score model based on BMPC ≥ 30% (1 point) and immunoparesis (1 point).
We identified patients with score 0 (20%) as low-risk (TTT at 60 months 0%), with score 1 (61%) as intermediate risk (TTT at 60 months 61%) and with score 2 as high-risk (TTT at 60 months 22%, p<0.0001).
Conclusion: We confirmed BMPC and immunoparesis as strong predictors of outcome in sMM.

Disclosures
No relevant conflicts of interest to declare.
Different Levels of Alkaline Phospatase (ALP) in Multiple Myeloma (MM) and SOLID Tumors with BONE Lesions
Authors: Annibali Ombretta , Petrucci Maria Teresa , Santini Daniele , Lisi Elisabetta , Mariani MArco , Pisani Francesco , Bongarzoni Velia , Venditti Olga , Rago Angela , Cerchiara Elisabetta , Fiorini Alessia , Franceschini Luca , Felici Stefano , Russano MArco , PAntano Francesco , Avvisati Giuseppe 
Published: October 2019

Abstract
Background: alkaline phosphatase (ALP) is an enzyme localized in different tissues and its levels may increase in diseases with skeletal involvement.
Aim: To compare ALP levels in patients with Multiple Myeloma and osteolytic lesions, and in patients with solid tumors (breast, prostate, lung, stomach, kidney and colon) and osteolytic, osteoblastic and mixed (osteolytic and osteoblastic) bone metastatic lesions.
Patients and Methods: From 1991 we collected 400 patients with MM and 308 patients with solid cancer with bone involvement. We grouped patients according to the metastases type (osteolytic, osteoblastic mixed), number of metastasis (1, 2-3, more than 3), ISS and D&S Stage. Among patients with solid tumors 45% had lytic lesions, while 31,5% had osteoblastic lesions and 23,5% had mixed lesions. Comparing MM vs. bone metastasis from solid cancer respectively 64.5% vs. 59% had normal ALP values, while 29% vs. 28% had <2xUNL, and 6.5% vs. 13% had >2UNL values of ALP. Patients with MM had significantly lower ALP values when compared to patients with osteoblastic bone lesions (P<0.05). In particular, comparing ALP levels of MM patients with the levels observed in cancer patients with >3 osteoblastic lesions difference is even more significant (P<0.01). Moreover, patients with Breast and Prostate cancer had ALP values significantly higher than MM pts (P<0.05 and P<0.05 respectively).In the group of patients with solid tumors, ALP levels were significantly higher in those with >3 osteoblastic lesions than in those with only osteolytic lesions (P<0.05).
Conclusion: These preliminary results indicate that ALP should be part of the initial work up in pts with bone lesions. This simple and cheap test, if normal or reduced in presence of osteolytic bone lesions, suggests an initial complete protein study including serum and urine protein electrophoresis associated to Bone Marrow aspirate, in case of the presence of a paraprotein.

Disclosures
Petrucci: Celgene, Janssen-Cilag, Amgen, Mundipharma, BMS: Honoraria.
Hyper-Active Non-Homologous End Joining Selects for Synthetic Lethality Resistant and Pathological Hematopoietic Stem Cells
Authors: Du Wei , Amarachintha Surya , Andrew Wilson , Pang Qishen 
Published: October 2019

Abstract
The prominent role of Fanconi anemia (FA) proteins involves homologous recombination (HR) repair. Poly[ADP-ribose] polymerase1 (PARP1) functions in multiple cellular processes including DNA repair and PARP inhibition is an emerging targeted therapy for cancer patients deficient in HR. Here we show that PARP1 activation in hematopoietic stem and progenitor cells (HSPCs) in response to genotoxic or oxidative stress attenuates HSPC exhaustion. Mechanistically, PARP1 controls the balance between HR and non-homologous end joining (NHEJ) in double strand break (DSB) repair by preventing excessive NHEJ. Disruption of the FA core complex skews PARP1 function in DSB repair and led to hyper-active NHEJ in Fanca-/- or Fancc-/- HSPCs. Re-expression of PARP1 rescues the hyper-active NHEJ phenotype in Brca1-/-Parp1-/- but less effective in Fanca-/-Parp1-/- cells. Inhibition of NHEJ prevents myeloid/erythroid pathologies associated with "synthetic lethality" and reduces human engraftment. Our results suggest that hyper-active NHEJ may select for "synthetic lethality" resistant and pathological HSPCs.

Disclosures
No relevant conflicts of interest to declare.
Stem Cell Mobilization Chemotherapy with Gemcitabine Is Effective and Safe in Myeloma Patients with Bortezomib Induced Neurotoxicity
Authors: Mueller Beatrice U. , Keller Sandra , Seipel Katja , Mansouri Taleghani Behrouz , Pabst Thomas 
Published: October 2019

Abstract
Background: Vinorelbine chemotherapy with G-CSF stimulation is a widely applied non-myelosuppressive mobilization regimen in Switzerland for myeloma patients, but its neurotoxic potential limits its use in patients with bortezomib induced polyneuropathy. Methods: In this single-center study, we alternatively evaluated safety and effectiveness of gemcitabine chemotherapy with G-CSF for mobilization of autologous stem cells. Results: Between 03/2012 and 02/2013, all bortezomib pretreated myeloma patients planned to undergo first-line high-dose melphalan chemotherapy received a single dose of 1250mg/m2 gemcitabine, with G-CSF started on day four. The 24 patients in this study have received a median of four cycles of bortezomib-dexamethason based induction. Bortezomib-caused polyneuropathy was identified in 21 patients (88%) by clinical evaluation and a standardized questionnaire. Administration of gemcitabine mobilization did not induce novel or aggravate preexisting neuropathy. Stem cell mobilization was successful in all 24 patients, with a single day of apheresis being sufficient in 19 patients (78%). The median yield was 9.51 x 106 CD34+ cells/kg. Stem collection could be accomplished at day 8 in 67%. Conclusion: Our data suggest that single-dose gemcitabine together with G-CSF is an effective mobilization regimen in myeloma patients and a safe alternative non-myelosuppressive mobilization chemotherapy for myeloma patients with bortezomib induced polyneuropathy.

Disclosures
No relevant conflicts of interest to declare.
Seroprevalence for Toxoplasma Gondii in Allogeneic Hematopoietic Stem Cell Transplantation Recipients
Authors: Atilla Erden , Ataca Pinar , Balli Sevinc , Isikoglu Baris , Topcuoglu Pervin , Ozcan Muhit , Akan Hamdi 
Published: October 2019

Abstract
Introduction:
Toxoplasmosis is a rare but serious and life-threatening infection following Allogeneic Stem Cell Transplantation (Allo-HSCT). The main cause of toxoplasmosis is the reactivation of latent infection in pre-transplant seropositive patients. In our study, our aim is to present toxoplasma gondii seroprevalence in our center.
Patients and Methods:
A total of 251 allogeneic stem cell transplant recipient were evaluated retrospectively from 1998 to 2015. During Allo-HSCT preparation procedures all recepients were serologically tested. Donor serology records were not adequate. Toxoplasma gondii-specific IgG and IgM antibodies were determined by Chemiluminescence by ARCHITECT lab analyzers using commercially available kits (Abbott, USA). All patients were followed for reactivation.
Results:
Toxoplasma IgG positivity was detected in 51 of patients (20.3%). There was no statistically difference detected between related or unrelated transplants. The mean age of the group was 36 (range 14-71). 144 recipients were male (57%). 208 (83%) patients were transplanted for malign hematological diseases. The majority of patients were grafted from full-matched related donors (173, 69%). The most common source of stem cell was peripheral blood in 192 patients (77%) followed by bone-marrow in 51 patients (51%), bone-marrow plus peripheral blood in 7 patients (2.8%) and cord in 1 patient (0.4%). 192 (77%) patients received myeloablative conditioning regimens. All patients received prophylactic twice weekly trimethoprim/sulfamethoxazole (TMP/SMX) (160/800mg PO daily) after engraftment. Two patients had clinical symptoms of toxoplasmosis however they were seronegative in serology. None of the patients developed an active toxoplasmosis after Allo-HSCT.
Conclusion:
Since less than 30% seropositivity defined as low rate, our institution had low rate seropositivity for Toxoplasma Gondii in Allo-HSCT recipients. Routine use of TMP/SMX prophylaxis and difficulties in diagnosis of toxoplasmosis may lead to low frequency active toxoplasmosis.

Disclosures
No relevant conflicts of interest to declare.
Outpatient Hematopoietic Grafting in Patients with Multiple Sclerosis Employing Autologous Non-Cryopreserved Peripheral Blood Stem Cells: A Feasibility Study
Authors: Ruiz-Arguelles Guillermo J. , León-Peña Andrés , Medina-Ceballos Emilio , Ruiz-Arguelles Alejandro , Ruiz-Delgado Manuel A , Ruiz-Delgado Rodrigo J , Ruiz-Reyes Guillermo , Priesca-Marin Manuel , Ruiz-Delgado Guillermo J 
Published: October 2019

Abstract
Background:
Multiple sclerosis (MS) is a chronic, inflammatory, debilitating disease that causes destruction of central nervous system (CNS) myelin, with varying degrees of axonal damage. With the goalofd re-setting the immune system, autologous hematopoietic stem cell transplantations (HSCT) have been done in patients with MS since 1996 and more than 700 HSCTs have been performed around the world. The risk of transplant related mortality in HSCT for MS has declined over the past years.
Material and methods:
Consecutive patients with MS were autografted in a single center using: Hematopoietic stem cells (HSC) were mobilized with cyclophosphamide (Cy), 3 gr/m2 and G-CSF, the procedure was conducted on outpatient basis employing peripheral blood non-frozen HSC and conditioning with high-dose Cy (100 mg/Kg) and post-transplant G-CSF and rituximab. Antibiotics, antimycotics and antivirals were given orally.
Results:
Thirteen patients with MS were prospectively accrued in the study. There were 7 females and 6 males. Median age was 48 years, range 24 to 65. The expanded disability status scale (EDSS) score of these patients had a median of 5 points (range 1 to 6). All the autografts were started on an outpatient basis and two persons were admitted to the hospital during the procedure (persistent nausea/vomiting and neutropenic fever); they stayed in the hospital for 48 hours. In order to obtain a minimum of 1 x106 viable CD34+ cells/Kg, one to four apheresis were done (median 1). The total number of viable CD34+ cells infused to the patients ranged between 1 and 9.6x106 (median 3.1). Patients recovered above 0.5 x109/L absolute granulocytes on median day 9 (range 6 to 12). No individuals needed transfusions of red blood cells nor platelets transfusions. There were no transplant-related deaths and the 23-month overall survival of the autografted patients is 100%. Median cost of the procedure was 30 000 USD. In 8 persons the EDSS was assessed three months after the graft; it diminished from a median of 4.5 to a median of 2.5. In 5 patients, the three months re-assessment of the EDSS has not been possible as a result of the time elapsed after the autograft.
Discussion:
These data indicate that it is possible to conduct autotrasplants for patients with MS employing a simplification of the conventional procedures by means of non-frozen peripheral blood stem cells and outpatient conduction. Additional information is needed to asses the efficacy of these procedures in the treatment of patients with MS.

Disclosures
No relevant conflicts of interest to declare.