bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bone Marrow-Derived Mesenchymal Stem Cells Overexpressing Akt1 Protect Against ConA-Induced Liver Injury in Mice
Authors: Zhou Lukun , Liu Shuang , Lu Chuanyi M. , Yao Jianfeng , Shen Yuyan , Yang Xin , Feng Sizhou , Han Mingzhe 
Published: October 2019

Abstract
Liver injury associated with veno-occlusive disease and graft-versus-host disease (GVHD) is a frequent and severe complication of hematopoietic stem cell transplantation, and remains an important cause of transplant-related mortality. Bone marrow derived mesenchymal stem cells (MSCs) have been evaluated for the prevention and treatment of refractory GVHD. However, poor cell viability has limited the therapeutic capacity of mesenchymal stromal cell therapy in vivo. In this study, we genetically engineered C57BL/6 mouse bone marrow MSCs using ex vivo retroviral transduction to overexpress Akt1, a serine threonine kinase and pro-survival signal protein, and tested the hypothesis that Akt1-expressing MSCs (Akt1-MSCs) are more resistant to apoptosis and can ameliorate acute liver injury induced by concanavalin A (ConA) in BALB/c mice. Cell proliferation and apoptosis analyses showed that, under both regular culture and high concentration IFN-γ (100 ng/mL) stimulation conditions, Akt1-GFP-MSCs had proliferation and survival (anti-apoptotic) advantages with down-regulated apoptosis pathways, compared to control GFP-MSCs. Twenty-four hours after receiving lethal dose of ConA (40 mg/kg, intravenous) (N=10 each group), no mouse survived, with or without 1x106 Akt1-MSCs or GFP-MSCs administration (intravenous); however, 3 and 1 survived in the 5×106 Akt1-MSCs group and 5×106 GFP-MSCs groups, respectively. In subsequent sub-lethal dose ConA (20 mg/kg) experiments, compared to GFP-MSCs, mice received Akt1-MSCs administration had significantly lower serum AST, ALT, TNF-α and IFN-γ levels and less histopathological abnormalities. In addition, Akt1-MSCs treated mice had significantly higher serum concentrations of IL-10, vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF). In vivo imaging showed that, hepatic fluorescence signal in sub-lethal ConA+Akt1-MSCs group was significantly stronger than ConA+GFP-MSCs group on day 0, and persisted up to 14 days, whereas the signal in ConA+GFP-MSCs, Akt1-MSCs and GFP-MSCs groups was negligible on both day 7 and day 14. Thus, bone marrow derived MSCs genetically enhanced with Akt1 had survival advantage in vitro and in vivo, and have the potential to be a potent therapy for prevention and amelioration of GVHD-associated liver impairment. Further translational pre-clinical studies are ongoing to further determine the efficacy, dosage and timing of administration of Akt1-MSCs in animal models.

Disclosures
No relevant conflicts of interest to declare.
5-Aza-2'-Deoxycytidine Promotes Cytotoxic Effect of CART-Cells on Leukaemia Cells
Authors: Dolnikov Alla , Rossi Swapna , Xu Ning , Klamer Guy , Shen Sylvie , Micklethwaite Kenneth P. , Lock Richard B , O'Brien Tracey 
Published: October 2019

Abstract
T cells modified to express CD19-specific chimeric antigen receptors (CAR) have shown anti-tumour efficacy in early phase clinical trials in patients with relapsed and refractory B-cell malignancies. In addition to direct cytotoxicity, chemotherapeutic drugs can have an immunomodulatory effect both through enhancing the tumour-specific immune response and increasing the tumour’s susceptibility to immune mediated destruction. Hence, combining immunomodulatory chemotherapy and CAR T-cells is an attractive approach for eliminating tumours, particularly in advanced stages. 5-aza-2'-deoxycytidine (5-AZA) is a hypomethylating agent that induces terminal differentiation, senescence or apoptosis in haematological malignancies. Here, we have explored a CAR-based immunotherapy combined with 5-AZA to maximise the effect of the CAR T-cells against CD19+ B-cell leukaemia. A second generation CAR including CD3zeta and the CD28 co-stimulatory domain was cloned into the PiggyBac-transposon vector and was used to generate CAR19 -T cells. Cord blood -derived mononuclear cells (MNC) were transfected with CAR19-transposon/transposase plasmids and expanded with CD3/28 beads for 2 weeks in the presence of 20ng/ml IL2 and 10ng/ml IL7. CAR19 T-cells efficiently induced cytolysis of CD19+ leukaemia cells in vitro and exhibited anti-tumour activity in vivo in a xenograft mouse model of leukaemia. Pre-treatment with 5-AZA produced greater leukaemia cell cytolysis in vitro and maximised anti-tumour activity of CAR19 T-cells in vivo against xenograft primary leukaemia compared to 5-AZA or CAR19 T-cells alone. In vitro analysis revealed that pre-treatment with 5-AZA up-regulates CD19 expression in leukaemia cells and improves CAR19 T-cell recognition of target cells increasing the formation of effector/ target cell conjugates and target cell cytolysis. Therefore using 5-AZA pre-treatment can be particularly useful for B-cell leukaemias with reduced expression of CD19. We have also demonstrated that pre-treatment of target cells with 5-AZA potentiates the effect of CAR19 T-cells used at low dose or low effector:target (E:T) suggesting that even small numbers of CAR19 T-cells can mediate a potent antitumor effect when combined with 5-AZA pre-treatment of target cells. This is particularly important for patients receiving limited numbers of CAR T-cells or for patients with large leukaemic burden. In addition, we speculate that the enhanced cellular cytotoxicity produced by 5-AZA-conditioning may allow the infusion of decreased numbers of CAR19 T-cells.

Disclosures
No relevant conflicts of interest to declare.
What Can We Do for the Unrelated Donors of Hematopoietic Stem Cell to Lessen Their Agony?
Authors: Lee Mark , Na Jung Hwa , Jang Hyung In , Jang Jin Ho , Lee Jongchan , Lyu Chuhl Joo , Kim Sung-Yong 
Published: October 2019

Abstract
Objectives: The study purposes are as follows. What are the incidences of adverse events (AEs) after hematopoietic stem cell donations among the unrelated donors registered in Korea Marrow Donor Program (KMDP)? What donor factors have influence on their general discomfort and the limitation of daily activity at the time of stem cell collection? What prohibits the donors from donating stem cell repetitively as a Good Samaritan?
Methods: 1,666 consecutive unrelated donors were registered at KMDP from November 2007 to October 2013. General discomfort and pain were assessed using a visual analog scale (numerical measurement, 0 to 10) and other donor symptoms were assessed by making either yes or no reply to each item. The limitation of daily activity was assessed by numerical measurement (0 to 10 scales), 10 being the worst activity. The donor’s intention of repetitive donation was assessed by yes or no reply. The donor symptoms and the intention of repetitive donation were assessed within 48 hours after stem cell collection, and then at 1 week, 4 weeks, and 4 months thereafter.
Results: There were 1,292 males and 374 females. 1,525 donors underwent peripheral blood stem cells (PBSC) collection, 100 bone marrow (BM) harvest and 40 leukapheresis for donor leukocyte infusion (DLI). All of 1,525 PBSC donors received granulocyte-colony stimulating factor (G-CSF). 76 donors underwent central line insertion for PB cells collection. The degree of general discomfort and the limitation of daily activity were analyzed in variables of sex, stem cell source, G-CSF administration, collection method, collection period and the intention of repetitive donation. A multiple linear regression analysis showed that general discomfort at the time of stem cell collection was significantly influenced by sex and collection period (P<0.05). The limitation of daily activity at the time of stem cell collection was significantly influenced by sex, stem cell source, and collection method (P<0.05). A multiple logistic regression analysis showed that the intention of repetitive donation at the time of stem cell collection and thereafter was significantly influenced by sex only. At least 80% of stem cell donors had the intention of repetitive donation.
Compared to the donors using peripheral line, the donors using central line for PB cells collection suffered more frequently from various AEs: C-line site discomfort, hands and feet numbness, perioral numbness, dizziness, chills, nausea/vomiting and abdominal pain.
Conclusions: Stem cell collection via peripheral line is recommended to alleviate the physical stress and to improve daily activity and to encourage repetitive donation, and female donors are more sensitive to stem cell collection than male donors.

Disclosures
No relevant conflicts of interest to declare.
Cryostem: Establishment of a National Thematic Collection of Biological Samples Pre- and Post-Allogeneic Hematologic Stem Cell Transplantation for the Study of Graft-Versus-Host Disease
Authors: Peffault de Latour Regis , Fontenille Claire , Yakoub-Agha Ibrahim , Calmels Boris 
Published: October 2019

Abstract
Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) is the only curative treatment for many patients with hematologic disease. Unfortunately, the transplanted immune system can reacts against the patient, inducing Graft-versus-Host Disease (GvHD). GvHD is the leading cause of treatment related mortality and morbidity following HSCT and is expected to increase in the forthcoming years. Pathophysiology and long-term determinants of GvHD are still largely unknown.
The French Society for Stem Cell Transplantation and Cell Therapy (SFGM-TC) launched the CryoStem project in 2011 with the goal of establishing a national biobank on GvHD. CryoStem has been selected and funded to the extent of 5,5 millions € over a period of 9 years by the French Government’s Investissements d’Avenir program. As of today, CryoStem brings together 33 HSCT Units and 23 Biological Resources Centers that actively contribute to a networked, prospective, longitudinal and standardized peripheral blood samples collection with concomitant well documented clinical data.
Patients sampling is scheduled before and after HSCT, depending on the occurrence or not of acute and/or chronic GvHD. Three types of pre-analytical products are processed and stored, according to standardized techniques: viable cells in DMSO, dried cell pellets and plasma. Samples and clinical data are anonymized and centralized in CryoStem-dedicated web-based database application.
Recruitment started in July 2012: currently, about 1,590 patients and 690 related donors have been included. So far, 3,820 blood samples have been processed, corresponding to 48,400 frozen pre-analytical samples. In order to guarantee the quality of frozen samples, an ISO9001 certification of the network is ongoing.
National projects are the only way to acquire the large number of samples needed for statistically significant results in a reasonable time. The collection is on embargo until further notice and the first call for scientific projects is expected to be launched in December 2014.
CryoStem is a crucial step toward progress in human GvHD research: it will allow collaborative studies between leading French and international research organizations and foster technology transfer with potential industrial players whenever possible to eventually improve the translational research on GvHD.

Disclosures
No relevant conflicts of interest to declare.
Rituximab in the Treatment of Epstein-Barr Virus Infection after hematopoietic stem cell Transplantation in Children
Authors: Peng Zhiyong , Li Chunfu , He Yuelin 
Published: October 2019

Abstract
OBJECTIVE: To investigate the efficacy and safety of Rituximab on Epstein-Barr virus infection disease after allogeneic hematopoietic stem-cell transplantation (allo-HSCT) in children.
METHODS: A retrospective analysis was performed based on clinical data of 25 children with median age of 6 years, (3~14 years) diagnosed as EBV infection and received Rituximab from Jan. 2012 to Jun. 2014 in our center. Of them, 3 patients were diagnosed as post-transplant lymphoproliferative disorders (PTLD). All patients received Rituximab at dose of 375 mg/m2 once a week for 4 sequence weeks or discontinued therapy when EBV viral loads was <500 copies/mL two times consecutively. The complete remission (CR) was defined as EBV viral loads <500 copies/mL consecutively at least two time and without any related clinical symptoms. Side effects during infusion were evaluated by Common Terminology Criteria for Adverse Events.
RESULTS: The median EBV-infection time was on day 70 (18~200) after HSCT. Each of patients received a median of 2 (1~4) infusions. There were no severe side effects during the infusion of Rituximab. The cumulative complete remission (CR) rate were 88% in total. The CR rate was 56.0±9.9 %, 80.0±8.0%, 84.0±7.3%, and 88.0±6.5%, respectively, in 1st, 2nd, 3rd, and 4th week. Two patients died, one due to infections and the other died of hepatitis while receiving therapy. The CR rate was 100% in patients with PTLD (3/3).
CONCLUSIONS: These data suggest Rituximab is safe and effective when treating children with EBV infection disease after allo-HSCT.

Disclosures
No relevant conflicts of interest to declare.
Effect of Severe Hypoalbuminemia on Myelosuppression and Other Toxicities of High Dose Melphalan and Autologous Stem Cell Transplantation in AL Amyloidosis Patients
Authors: Meehan Robert , Seldin David , Sloan John Mark , Quillen Karen , Brauneis Dina , Shelton Anthony C , Sanchorawala Vaishali 
Published: October 2019

Abstract
Background: Treatment of AL amyloidosis with high dose intravenous melphalan followed by autologous stem cell transplant (HDM/SCT) is effective in inducing hematologic and clinical remissions associated with prolonged survival. The major toxicities are myelosuppression and GI side effects. Studies have shown that ~75% of melphalan in the blood is bound to plasma proteins, with ~25% free. We hypothesized that AL patients with severe nephrotic syndrome and profound hypoalbuminemia might have a higher fraction of free melphalan, a higher effective dose, and greater toxicity of treatment.
Methods: Patients with AL amyloidosis and severe hypoalbuminemia, defined as serum albumin level of < 2 g/dL, treated from 2011 to 2013, were studied retrospectively. The stem cell transplant database was queried for dose of HDM, treatment-related complications, and days of neutrophil and platelet engraftment after SCT.
Results: Of 71 patients with AL amyloidosis who underwent HDM/SCT between Jan 2011 and Dec 2013, 12 patients had severe hypoalbuminemia. Of these, 5 received full HDM at 200 mg/m2 and 7 received modified HDM at 140 mg/m2. All patients received GCSF mobilized peripheral blood stem cells following HDM, with a median stem cell dose of CD34+ cells 8.1 x 106/kg (range, 4.0 to 12.2). The median time to engraftment of neutrophils was 11 days, and not statistically different based upon melphalan dose. The median time to platelet engraftment was 13 days, and also did not differ significantly by dose. These times were similar to controls without severe hypoalbuminemia. Grade 4 toxicities were observed in 2 of 7 patients with modified HDM/SCT and 1 of 5 patients with full HDM/SCT.
Conclusions: These data suggest that patients with severe hypoalbuminemia do not have more prolonged myelosuppression or increased non-hematologic toxicities compared to other patients. In this retrospective study, we did not measure free melphalan concentrations in the blood. However, these data suggest that patients with severe hypoalbuminemia do not require adjustment of melphalan dosing.

Disclosures
No relevant conflicts of interest to declare.
Successful Unrelated Peripheral Blood Stem Cell Transplantation from Unrelated Donor for a Concomitant Paroxysmal Nocturnal Hemoglobinuria and Heterozygous α-Thalassemia Patient
Authors: Wu Xuedong , Wen Jianyun , Liu Xuan , Wang Yanhua , Lu Linglong , Li Na , Li Chunfu 
Published: October 2019

Abstract
Background: Paroxysmal nocturnal haemoglobinuria (PNH) is a rare acquired clonal hematopoietic stem cell (HSC) disorder that manifests as hemolytic anemia, venous thrombosis, and deficient hematopoiesis. PNH concomitant with inherited hemolytic anemia has been rarely reported. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is considered the only curative therapeutic measure. Here, We present our first allo-HSCT form unrelated donor for a 23-year-old patient with PNH and heterozygous α-thalassemia.
Case report: Flow cytometry of the patient's peripheral blood demonstrated that 7.9% and 11.9% of the erythrocytes were CD59 and CD55 deficient, respectively. Thalassemia gene analysis revealed heterozygous α-thalassemia. The patient was diagnosed with PNH and heterozygous α-thalassemia, and became transfusion dependent 1 year later and was admitted to our hospital in March 2011. Conditioning was consisited of Cyclophosphamide (d-10 to d-9), Fludarabine(d-8 to d-4), Busulfan (d-7 to d-5), Thiotepa (d-4) and ATG-F (d-3 to d-1). Stem cell source was peripheral blood stem cells (PBSC) harvested on the 16th October 2011 from a HLA-matched (8/8) unrelated donor. The infused mononuclear cell (MNC) and CD34+cell does were 8×08/kg and 1.38×106/kg body weight of the recipient, respecitively.The time to platelets engraftment and leukocyte engraftment was 21 and 25 days,respecitively.The patient did not accoured acute graft versus host disease (aGVHD). Out-patient were followed up for 2 years, the patient's liver and kidney function, urine which showed normal, as well as reviewed the implant remains for independent plantentry.
Conclusions: The present case suggests that allo-HSCT can be used to cure patient with PNH and heterozygous α-thalassemia. Allo-HCT from unrelated donors is a new and potentially curative option for patients with PNH and inherited hemolytic anemia.

Disclosures
No relevant conflicts of interest to declare.
Human C1q Deficiency and Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Olsson Richard , Ringden Olle , Truedsson Lennart , Åhlin Anders 
Published: October 2019

Abstract
Human C1q deficiency is a rare disorder associated with systemic lupus erythematosus (SLE) and increased susceptibility to severe bacterial infections with mainly encapsulated organisms. Among patients with C1q deficiency there is a great variation in the severity of symptoms, however, the majority of patients requires comprehensive medical therapy and some develop treatment resistant disease. Since C1q is produced by monocytes it has been speculated that allogeneic hematopoietic stem cell transplantation (allo-HSCT) may cure these patients. At our center, we have so far treated five patients with C1q deficiency. In three cases, SLE symptoms remained relatively mild after the initiation of medical therapy, but two patients developed treatment resistant SLE and we decided to pursue with allo-HSCT. Here, we report an unrelated allo-HSCT in a 9-year-old boy who suffered from refractory SLE of the central nervous system and a related allo-HSCT in a 12-year-old girl with refractory SLE. Both patients restored C1q production post-transplant, and the severity of their SLE was attenuated. The boy developed post-transplant lymphoproliferative disease, which resolved after rituximab and donor lymphocyte infusions (DLI). Unfortunately, the DLIs induced cortisone-resistant severe gastrointestinal graft-versus-host disease, and he died from multiple organ failure four months post-transplant. The girl is doing well 21 months post-transplant, and clinically all signs of SLE have resolved. We believe that allo-HSCT is a promising treatment in patients with C1q deficiency and severe SLE, which needs further investigation.

Disclosures
No relevant conflicts of interest to declare.
High Glucose/Thrombin-Induced Endothelial Inflammation Via Microrna-146a and Nox4 Regulation
Authors: Peng Ching-Tien , Lo Wan Yu , Wang Huang Joe 
Published: October 2019

Abstract
Diabetes is associated with hyperglycemia and increased thrombin generation. It is unknown whether high glucose (HG)/thrombin can modulate the expression of NAPDH oxidase (Nox) subtypes in human aortic endothelial cells (HAECs). Besides, we investigate whether miR-146a is involved in endothelial cell inflammation. We observed that HG (25 mmol/l) exerted a synergistic effect with thrombin (2 U/ml) for induction of Nox4 mRNA level in HAECs. The increased Nox4 mRNA was associated with increased Nox4 protein and ROS production. We also demonstrated that HG/thrombin treatment increased interleukin-8 and interleukin-6 protein levels. Besides, HG/thrombin treatment caused an 11.43-fold increase of THP-1 adhesion to HAECs. In Silico analysis identified homology between miR-146a and the 3’-UTR of the human Nox4 mRNA, suggesting a potential regulation of Nox4 by miR-146a. Furthermore, HG/thrombin treatment decreased miR146a expression to 58% of the control, indicating an impaired feedback restrain of HG/thrombin-induced endothelial inflammation. MiR-146a mimic transfection prevented HG/thrombin-induced upregulation of Nox4 mRNA, Nox4 protein, and ROS generation. In addition, inflammatory phenotypes were attenuated in miR-146a mimic-transfected HAECs. In conclusion, miR-146a is involved in the regulation of endothelial inflammation via modulation of Nox4 in an in-vitro milieu mimicking diabetic atherothrombosis.

Disclosures
No relevant conflicts of interest to declare.
Evidence of Defibrotide Internalization and Its Protective Effect in a Hepatic Endothelial in Vitro model
Authors: Diaz-Ricart Maribel , Palomo Marta , Mir Enric , Rovira Montserrat , Escolar Gines , Carreras Enric 
Published: October 2019

Abstract
INTRODUCTION: Defibrotide (DF) interferes with several steps of the coagulation-inflammation cycle. Last October (2013), DF received EMA authorization for its use in the treatment of severe hepatic veno-occlusive disease (VOD) after hematopoietic cell transplantation (HCT). Since endothelial dysfunction has been associated with the development of VOD, among other HCT complications, we decided to explore the mechanisms of action of DF in a hepatic endothelial in vitromodel.
OBJECTIVES: To investigate the interaction of DF with a hepatic endothelial cell line from human origin (SK-HEP-1), the mechanisms involved in its traffic and its potential protective effect on fludarabine induced apoptotic mechanisms.
METHODS: SK-HEP-1 cells were exposed to DF (20µg/ml), previously labelled with ULYSIS® Nucleic Acid Labeling Kit, for up to 24 hours. Confocal microscopy (CM, Leica TCS SP5) was used to localize DF into the different cell compartments. Additional experiments were performed to assess colocalization of DF with different endocytosis pathways, by using specific antibodies. MTT assay to measure cell viability was performed in SK-HEP-1 cells after incubation with DF at different concentrations (10 to 200µg/mL, for 24h) followed by fludarabine exposure (at 20µg/mL, for 24h).
RESULTS: CM analysis revealed a significant interaction of DF with endothelial cell membranes followed by internalization and redistribution to the cytoplasm. Mechanisms involved seem to be dependent on endosomes and cytoskeletal assembly. DF did not reach the cell nucleus even after 24h of exposure. In addition, MTT assay demonstrated the protective effect of DF (at concentrations >20µg/mL) on the fludarabine damage on endothelial cells.
CONCLUSIONS: Our studies show that DF interacts with endothelial cell membranes, becoming internalized and redistributed into the endothelial cell cytoplasm without evidence in the nucleus. Moreover, DF protects hepatic endothelial cells from the toxic effect of fludarabine. Our findings may contribute to a better understanding of the precise mechanisms of action of DF as a therapeutic and potential preventive agent on the endothelial damage underlying different pathological situations.
Partially supported by grant SAF2011-28214 (Spanish MINECO and FEDER).

Disclosures
Diaz-Ricart: Gentium / Jazz Pharmaceuticals: Research Funding. Palomo:Gentium / Jazz Pharmaceuticals: Research Funding. Mir:Gentium / Jazz Pharmaceuticals: Research Funding. Carreras:Gentium / Jazz Pharmaceuticals: Research Funding.