bluuebunny/crossref_metadata_2025_split huggingface dataset:
Osteogenic/Adipogenic Imbalance in Bone Marrow Nestin+ mesenchymal Stem Cells in Patients with Chronic Graft-Versus-Host Disease
Authors: Qi Hanzhou , Zhang Haiyan , Yang Kaibo , Chen Yanqiu , Jin Hua , Liu Qifa 
Published: November 2019

Background: Chronic graft-versus-host disease (cGVHD) is a major cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The roles of bone marrow niche in cGVHD pathogenesis have gained increasing attention in recent years. Nestin+ mesenchymal stem cells (MSCs) is an important component of bone marrow niche. However, the potential implication of Nestin+ MSCs in the pathophysiology of cGVHD has not been completely clarified.
Methods: A total of 68 patients with hematologic malignancies who underwent allo-HSCT at Nanfang Hospital between April 2016 and October 2018 were enrolled in this experimental study. We analyzed expanded Nestin+ MSCs from patients with cGVHD and compared them with patients without cGVHD. The diagnosis and grade of cGVHD was made at the time of sample collection according to National Institutes of Health criteria (Howard M.S. et. al. BBMT 2015). The results were compared using one-way ANOVA and unpaired two-tailed Student t-test. Statistical significance was defined as P value of <.05.
Result:
The Nestin+ MSCs from both groups showed similar morphology, immunophenotype, proliferation, and apoptosis. However, the adipogenic differentiation capacity of Nestin+ MSCs in patients with cGVHD was significantly reduced compared with patients without cGVHD (relative expression of PPARγ 2.22±0.27, and 6.82±0.87, respectively, P<0.05). The osteogenic differentiation capacity was significantly increased in patients with cGVHD (relative expression of RUNX2 3.84±0.38, and 1.95±0.52, respectively, P<0.05)(Figure 1A,B). These abnormal differentiations were more significant in patients with moderate/severe cGVHD. Furthermore, β-catenin phosphorylation decreased and nuclear β-catenin increased in the Nestin+ MSCs of cGVHD patients(Figure 1 C).
Conclusion: These results demonstrate that Nestin+ MSCs from cGVHD patients had abnormal differentiation characterized by decreased adipogenic differentiation capacity and enhanced osteogenic capacity. The reduction of phosphorylation of β-catenin play an important role in these abnormal differentiations.

Disclosures
No relevant conflicts of interest to declare.
Epidermal Growth Factor Stimulates Exosomal microRNA-21 Derived from Mesenchymal Stem Cells to Ameliorate aGVHD By Modulating Regulatory T Cells
Authors: Chen Minmin , Zhu Haojie , Lan Lingqiong , Zhang Yuxin , Chen Qiuru , Zeng Yanling , Luo Xiaofeng , Ren Jinhua , Chen Zhizhe , Hu Jianda , Yang Ting 
Published: November 2019

Objective: Several microRNAs have been revealed to be involved in the pathophysiological process of GVHD. This study aimed to characterize the functional relevance of epidermal growth factor (EGF)-stimulated microRNA-21 (miR-21) in the bone marrow-derived mesenchymal stem cells (BMSCs) in a mouse model of aGVHD.
Methods: BMSCs and regulatory T cells (Tregs) were isolated and cultured. The effects of miR-21 knockdown or overexpression and EGF on cell activities of BMSCs as well as the expression of PTEN, Foxp3, AKT phosphorylation and extent of c-jun phosphorylation were examined by gain- and loss-of-function approaches. The efficacy and safety of EGF were further assessed in the mouse model of aGVHD.
Results: Overexpression of miR-21 promoted the proliferation, invasion and migration of BMSCs. Besides, miR-21 in BMSCs-derived exosomes inhibited the expression of PTEN, enhanced AKT as well as GSK-3β phosphorylation and Foxp3 expression in Tregs. In addition, EGF enhanced c-jun phosphorylation to elevate the miR-21 expression. Furthermore, EGF significantly increased the effect of BMSCs in the mouse model of aGVHD, corresponding to reduced IFN-γ expression and organ damage. Moreover, the Foxp3 expression of Tregs in BMSCs-treated aGVHD mice was promoted by the treatment of EGF.
Conclusion: Taken together, EGF induced the BMSCs-derived exosomal miR-21 expression to enhance the expression of Foxp3 in Tregs, thereby improving the therapeutic effect of BMSCs on aGVHD.

Disclosures
No relevant conflicts of interest to declare.
Central Nervous System (CNS) Infiltration Assessment Using Cerebrospinal Fluid Flow Cytometry in Hematologic Malignancies: A Single Center Retrospective Analysis of Clinical Outcomes
Authors: Chapchap Eduardo Cerello , Feres Carolina , Bento Laiz Cameirão , Schimidell Daniela , Correia Rodolfo Patussi , Alexandre Anderson Marega , Nosawa Sonia Tsukasa , Pedro Eduardo de Carvalho , Vaz Andressa da Costa , Barroso Rodrigo De Souza , Duarte Carlos Augusto Senne , Bacal Nydia Strachman , Hamerschlak Nelson 
Published: November 2019

INTRODUCTION: The detection of central nervous system (CNS) disease in hematologic malignancies is important to guide optimal therapeutic approach, refine prognosis and understand patient`s unexplained neurologic symptoms. Newer flow cytometry (FC) techniques are emerging, also there are increasingly reports of higher accuracy than routine cytospin. Moreover, these are becoming incorporated more frequently in clinical work up practices. However, there is still uncertainty on clinical approach of CNS-positive patients, detected only by FC.
OBJECTIVE: To analyze accuracy and clinical outcomes of CNS disease by cytopsin or FC in patients with hematologic malignancies.
MATERIAL AND METHODS: FC cerebrospinal samples and medical charts of 84 consecutive patients evaluated for CNS infiltration by hematologic malignancies from January/2014 to December/2016 were reviewed. Statistical analysis were done with SPSS and STATA softwares.
RESULTS: Baseline patients characteristics were: male (62%), median age 53 years; non-hodgkin lymphoma (52%), Acute Lymphoblastic Leukemia (26%), Acute Myeloblastic Leukemia (15,5%), Multiple Myeloma (6,5%); CNS-positivity rates according to each technique were: Cytopsin-/FC- (71,4%), Cytospin+ (14,3%), Cytospin-/FC+ (14,3%); CNS-disease was detected by FC in 32,3%, while for cytospin was 16,7%. Overall survival was 71,4% and relapse rate 38,1% at 2,5 years of median follow-up. Relapsed (HR: 2,76 p0,023) and CNS-positivity (HR: 2,01 p0,037) patients were significantly associated with an inferior overall survival. Also, progression free survival (PFS) of Cytospin-/FC+ was significantly inferior than CNS-negative subgroup (HR: 2,93 p0,022).
CONCLUSION: FC sensitivity appears to be higher than classicaly cytospin methods to detect CNS disease, also CNS-positivity was associated with a worse prognosis, as well as in the subset of patients Cytospin-/FC+. Further studies with a more homogeneous cohort and larger sample sizes are needed to validate our findings.
Figure

Disclosures
No relevant conflicts of interest to declare.
Combination of Haploidentical Hematopoietic with Low-Dose ATG, Ptcy and Cord Blood Competitive Transplantation for Hematologic Malignancies
Authors: Chen Yiling , Li Xiaofan , Xu Zhenshu , Yuanzhong Chen , LI Nainong 
Published: November 2019

Objective: To evaluate the efficacy and safety of the combination of haploidentical hematopoietic with low-dose ATG, PTCy and cord blood competitive transplantation for hematologic malignancies.
Methods: This study was conducted as a retrospective review of medical records of 31 patients with hematologic malignancies who received a combination of haploidentical hematopoietic with low-dose ATG, PTCy and cord blood competitive transplantation at Fujian medical university union hospital, from November 2016 to April 2019.
Results: Among the 31 patients with hematologic malignancies, the median age was 22 (6-52) years, of which 17 were CR patients and 14 were No CR patients. The conditioning was modified FABuCy+ low-dose ATG (total dose 5mg/kg) based regimen. Cord blood units were selected based on the results of HLA typing and cell doses evaluated before freezing. Units with at least 5/10 matched HLA loci became the candidates. Prophylaxis for graft-versus host disease(GVHD) was by CsA(cyclosporine), MMF (mycophenolate mofetil) plus PTCy(post-transplant cyclophosphamide). The median values of absolute total nucleated cell counts were 123.0 (49.0-258.8) × 107 / kg in The haploidentical grafts and 2.5 (1.1-13.0 × 107 / kg in the cord blood units,respectively. The median Doses of CD34+ cells infused were 70.0 (11.8-297.6) × 105 / kg in the haploidentical grafts and 1.4 (0.1-13.0) × 105 / kg in the cord blood units, respectively. All patients attained complete engraftment , of which 19 were haploidentical engraftment and 12 were cord blood engraftment. The median durations of myeloid engraftment were 14 (12-37) days and 14.5 (10-48) days for Platelets. With a cumulative platelet engraftment incidence of 90.3%. During a median follow-up of 8.5 (1.3-30.7) months, the incidence of grade II-IV acute GVHD at 100-day was 47.5%, grade III-IV acute GVHD was 13.8%, respectally. The incidence of limited chronic GVHD at 1-year was 35.1%, severe chronic GVHD was 14.6 %, respectively. The estimated 1-year OS was 76.1%,DFS was 68.5% ,GRFS was 60.6%, NRM was 18.5% , RM was 6.7% , respectively. The estimated 1-year OS of CR patients and No CR patients were 92.9% and 55.0%(p=0.114), DFS were 83.6% and 51.9%(p=0.053),NRM were 7.1% and 31.2%(p=0.132),relapse were 10.0% and 30.4%(p=0.131), respectively.
Conclusion:The results suggested that the combination of haploidentical hematopoietic with low-dose ATG, PTCy and cord blood competitive transplantation may potentially improve the outcome of HSCT. It offers a transplant alternative for patients with hematologic malignancies who lack matching donors.

Disclosures
No relevant conflicts of interest to declare.
KIR-Genotype Mismatch, Donor KIR Locus 3DS1 Positive or 2DS3 Negative Facilitates a Positive Outcome of Allogeneic Hematopoietic Stem Cell Transplantation: A Single-Center Retrospective Study of 74 Patients
Authors: Zhang Yu , Chen Shaozhen , Ren Jinhua , Luo Xiaofeng , Hu Jianda , Yang Ting 
Published: November 2019

Objection: To investigate the potential influence of donor-recipient KIR genotype matching and KIR alleles on the prognosis of allogeneic hematopoietic stem cell transplantation (allo-HSCT).
Method :74 cases of AML patients who admitted and treated in the transplantation ward of our hospital from May 2015 to May 2019 were analyzed retrospectively. Binary Logistic regression and k-m survival curve were used to demonstrate the influence of donor age, blood group ABO compatibility, donor-recipient HLA matching, donor-recipient KIR mismatch model and donor KIR gene locus on the transplant outcome.
Result :Among the 74 patients with AML, 44 were male (59.5%) and 30 female (40.5%), with the median age of 24 years (1-59 years). 49 cases (66.2%) were in remission at the time of transplantation, and 25 (33.8%) not in remission. aGVHD occurred in 16 patients (21.6%) and recurrence occurred in 11 patients (14.9%). Multivariate analysis revealed that KIR-genotype matching was closely related to the increased risk of aGVHD. The incidence of aGVHD in the KIR mismatching group was significantly lower than that in the matching group (12% vs 31%, P=0.019), but no significant effects on survival and recurrence. Donor KIR 3DS1 negative had an increased risk of aGVHD compared with those with 3DS1 positive (28.2% vs 11.7%, P=0.033). The recurrence rate (RR) of donor KIR 2DS3 negative, even when the recipients with hematologic malignancies were not in remission at the time of HSCT, were much lower than those with 2DS3 positive (10% vs 66.7%, P=0.045). However, donor age, blood group ABO compatibility, donor-recipient HLA matching, and other donor KIR locus had no significant effect on OS, aGVHD and RR.
Conclusion: Thus, it is recommended to select the donor with KIR mismatching, KIR locus 3DS1 positive or 2DS3 negative to improve the outcome of allo-HSCT by reducing the risk of aGVHD or recurrence risk, which offer a clinically applicable donor selection strategy.

Disclosures
No relevant conflicts of interest to declare.
A Randomized Evaluation of Vinorelbine Versus Gemcitabine Chemotherapy Mobilization of Stem Cells in Myeloma Patients
Authors: Jeker Barbara , Farag Sarah , Novak Urban , Bacher Ulrike , Mansouri Taleghani Behrouz , Li Qiyu , Pabst Thomas 
Published: November 2019

Background: In myeloma patients, a non-myelosuppressive regimen combining a single dose of vinorelbine chemotherapy together with G-CSF is safe and effective to harvest peripheral blood progenitor and stem cells (PBSC) before autologous stem cell transplantation (ASCT). Considering its neurotoxic potential, vinorelbine may aggravate pre-existing bortezomib-induced polyneuropathy, and gemcitabine represents an alternative mobilization regimen. We have reported that monotherapy with gemcitabine together with G-CSF can be used as a promising alternative mobilization chemotherapy to vinorelbine, even more so as gemcitabine avoids neurotoxicity associated with vinorelbine
Methods: In this prospective phase-II study (ViGeM Trial; NCT# 02791373), we compared in a 1:1 randomization the efficacy and toxicity of vinorelbine or gemcitabine with G-CSF (and, if needed, plerixafor) in 130 evaluable myeloma patients in first remission. The objective of the study is to demonstrate the non-inferiority of gemcitabine versus vinorelbine. The primary endpoint is mobilization success, which is defined as a collected apheresis product comprising at least 6.0×106 CD34+ cells/kg b.w. at day 8 after chemotherapy (vinorelbine or gemcitabine) together with G-CSF in the absence of plerixafor.
Results: We observed successful CD34+ mobilization in 75% [95% confidence interval (CI): 63%-85%] of patients in the vinorelbine group versus 49% [95% CI: 36%-62%] with gemcitabine; the pre-specified non-inferiority margin -15% was not reached. More vinorelbine recipients achieved the collection goal in a single-day procedure (78% vs 60%). The median CD34+ yield was 11.4×106/kg b.w. in vinorelbine versus 8.7×106/kg in gemcitabine (P=0.001). At apheresis as well as day +100 following ASCT, polyneuropathy occurred more frequent in vinorelbine recipients. Special attention should be given to the fact that no patients with gemcitabine was observed with grade ≥ 3 polyneuropathy (Table 1). Finally, less patients in the vinorelbine group (1%) needed two apheresis days as compared to 14% in the gemcitabine group, respectively (P=0.007), whereas the use of rescue plerixafor was similar (11% versus 10%).
Conclusion: This prospective comparison indicated that non-inferiority of gemcitabine as compared to vinorelbine chemotherapy together with G-CSF stimulation regarding CD34+ mobilization was not reached. Polyneuropathy was aggravated in the vinorelbine cohort, but not in gemcitabine recipients. Our study allows individualization of CD34+ mobilization regimens in myeloma patients.

Disclosures
No relevant conflicts of interest to declare.

OffLabel Disclosure:
Vinorelbine and gemcitabine are used for stem cell mobilization in this study and, thus, off-label.
Anti-CD19-CAR T-Cell from Patients Relapsed after Allo-HSCT with a Low Level of Donor Chimerism Restored to Complete Donor Chimerism: Successful Cases and Analysis
Authors: Li Qing , Mu Juan , Yang Zhen-xing , Mou Nan , Wang Jia , Yuan Ji-jun , Jiang Yanyu , Juanxia Meng , Deng Qi 
Published: November 2019

Purpose: To study the efficacy of anti-CD19-CAR T-cell therapy and the changes of donor chimerism in CAR-T cells in vitro culture for patients who relapsed after allo-HSCT with a low level of donor chimerism.
Methods: Two patients relapsed after allo-HSCT with a low level of donor chimerism (11.22% and 16.08%). Their autologous PBMCs were selected for anti-CD19-CAR T-cell therapy because their sibling or unrelated donor could not provide PBMC cells. The adverse events (AEs) were observed after CAR T-cell infusion. The cytokine release syndrome (CRS), the expansion of anti-CD19-CAR T-cells, the DNA level of anti-CD19-CAR gene and the changes of the donor chimerism were detected after CAR T-cell infusion. Then we selected other 7 acute leukemia patients relapsed after allo-HSCT with a low level of donor chimerism to detect the changes of donor chimerism in their T cells and anti-CD19-CAR T-cells in vitro culture.
Results: Their notable AEs were grade 2 CRS. They had no aGVHD during anti-CD19-CAR T-cell therapy. They achieved CR 14 days after anti-CD19-CAR T-cell therapy with a restoration of complete donor chimerism (98.61% and 98.48%). To date, they had leukemia-free survival (LFS) and complete donor chimerism for 8 and 7 months. It was very interesting and fortunate that the CAR-T cells infused into the two patients were restorated to complete donor chimerism (99.2% and 99.4%). Then we found that complete donor chimerism were restored after 12 days culture in vitro in all the T cells and anti-CD19-CAR T-cells in the other 7 patients relapsed after allo-HSCT with a low level of donor chimerism.
Conclusions: Anti-CD19-CAR T-cells derived from patients relapsed after allo-HSCT with a low level of donor chimerism were effective for this salvage therapy and could restorate to complete donor chimerism after 12 days culture in vitro.

Disclosures
No relevant conflicts of interest to declare.
Clinical Impact of Enrollment of Clinical Trial in Multiple Myeloma; Single Center Experience
Authors: Ahn Seo-Yeon , Jung Sung-Hoon , Song Ga-Young , Yang Deok-Hwan , Ahn Jae-Sook , Kim Hyeoung-Joon , Lee Je-Jung 
Published: November 2019

Purpose : Many clinical trials have been conducted to prove the efficiacy of the new agrent or treatment regimen in patients with multiple myeloma (MM), but there was no analysis of the effect of enrollment in clinical trials on survival in patients with MM. Therefore, we evaluate the survival outcome in patients with multiple myeloma who participated in clinical trial.
Method: Retrospective data of 446 patients with newly diagnosed MM between January 2003 and December 2016 in Chonnam National University Hwasun Hospital were analyzed. Patients diagnosed with monoclonal gammopathy of undetermined significance, asymptomatic MM, and plasma cell leukemia were excluded. In addition, non-interventional study was not included as a clinical trial in this study.
Results: The median overall survival (OS) for the entire cohort was 62.5 months (95%CI 52.4-72), and median OS has gradually improved (46.5 months in 2003-2008 vs. 64.9 months in 2009-2014 vs. not reached in 2015-2016, P = 0.007). A total of 197 patients (44.2%) participated in the clinical trial and 7.4% participated in more than two clinical trials during the entire treatment period. Participation rates in clinical trials were significantly higher in transplant-eligible patients than transplant-ineligible patients (55.7% vs. 34.8%, P <0.001). There was no significant difference in OS according to the enrollment of clinical trial in entire patients (62.9 months vs. 64.9 months, P = 0.265). In transplant-eligible patients, patients who participated clinical trial showed improved OS than patients who did not participate clinical trial (77.0 months vs. 52.1 months, P = 0.058). However, in transplant-ineligible patients, no survival benefits were showed from participation in clinical trial (50.3 months vs. 66.0 months, P = 0.811)
Conclusion: This study showed survival benefit of enrollment of clinical trial in transplant-eligible patients. Further analysis is needed to analysis the effects of clinical trial in elderly patients.
Figure

Disclosures
No relevant conflicts of interest to declare.
Ex Vivo Activity of Immunotherapeutic Approaches Targeting CD38 Against Daratumumab-Resistant Multiple Myeloma Patient Samples
Authors: Karvouni Maria , Zhou Heyue , Wagner Arnika Kathleen , Ma Qiangzhong , Baloch Alamdar H. , Zhu Tong , Zhang Hong , Gilljam Mari , Lundqvist Andreas , Kaufmann Gunnar F. , Alici Evren 
Published: November 2019

Background: Multiple myeloma (MM) is a plasma cell malignancy that remains incurable. The identification of CD38, a transmembrane glycoprotein overexpressed on MM cells, led to the development of target-specific therapeutics such as the FDA approved monoclonal antibody (mAb) Daratumumab (DARA). Although a valuable treatment option for refractory/relapsed (R/R) MM patients, DARA has a limited response rate of below 50%, which highlights the clinical need for novel therapeutics.
Aims: Aiming to further exploit the therapeutic potential of CD38 in the MM setting, immunotherapies based on the novel anti-CD38 mAb CD38A2 were tested.
Methods: For the first approach, the CD38A2 mAb -that binds to a unique, distinct from DARA's, CD38 epitope- was conjugated with either the alkylating agent Duomycin (ADC-136) or the microtubulin binder Duostatin (ADC-129). The ADCs were compared to DARA, in cultures of primary MM cells from patients refractory to DARA treatment. In a second approach, a chimeric antigen receptor (CAR) consisting of the CD38A2 scFv and the intracellular domains of CD28 and CD3ζ was used to transduce primary T and NK cells from R/R MM patients. The functionality of the CAR-T and CAR-NK cells was assessed in cytotoxicity assays against autologous myeloma cells.
Results: ADC-136 demonstrated the most potent cytotoxicity against the MM cells with an IC50 of 6pM at day 6 following a single dose treatment. ADC-129 showed cell killing with an IC50 of 30pM, while DARA did not exhibit appreciable cytotoxicity. Regarding the cell therapy approach, patients' T and NK cells were effectively transduced, showing a CD38A2-CAR expression ranging between 11-68%. In functional assays, CAR-T and CAR-NK cells were assayed against autologous myeloma cells, where they exhibited an increase in target cell cytotoxicity, compared to the untransduced cells.
Summary/Conclusion: Altogether, our preliminary findings demonstrate that CD38 targeting using CD38A2-based immunotherapies could be a viable therapeutic approach in R/R MM patients previously exposed to DARA. Currently, an anti-CD38 CAR-T therapy based on CD38A2 is being evaluated in Phase 1 studies in R/R MM patients by Sorrento Therapeutics, Inc.

Disclosures
Zhou: Sorrento Therapeutics Inc: Employment, Equity Ownership. Ma:Sorrento Therapeutics Inc: Employment, Equity Ownership. Zhu:Sorrento Therapeutics Inc: Employment, Equity Ownership. Zhang:Sorrento Therapeutics Inc: Employment, Equity Ownership. Kaufmann:Sorrento Therapeutics, Inc.: Employment, Equity Ownership, Patents & Royalties.
The Efficacy of a Novel Oral Proteasome Inhibitor NNU546 in Multiple Myeloma
Authors: Yang Ye , Zhu Yongqiang , Gu Chunyan 
Published: November 2019

Proteasome inhibition demonstrates highly effective impact on multiple myeloma (MM) treatment. In this study we aimed to examine a novel well tolerated orally applicable proteasome inhibitor NNU546 and its hydrolyzed pharmacologically active form NNU219. NNU219 showed more selective inhibition to proteasome catalytic subunits and less off-target effect than bortezomib ex vivo. Furthermore, intravenous and oral administration of NNU219 and NNU546, respectively, led to a more sustained pharmacodynamic inhibition of proteasome activities compared with bortezomib. Intriguingly, NNU219 exhibited potential anti-MM activity on both MM cell lines and primary samples in vitro. The anti-MM activity of NNU219 was determined to be associated with induction of G2/M-phase arrest, as well as induction of apoptosis via activation of the caspase cascade and endoplasmic reticulum stress response. At well-tolerated doses, significant growth-inhibitory effects of NNU219 and NNU546 were observed in 3 different human MM xenograft mouse models and significant anti-MM activity was observed even in the presence of a bone marrow microenvironment. Taken together, these findings provided the basis for clinical trials of NNU546 to determine its potential as a candidate for MM treatment.

Disclosures
No relevant conflicts of interest to declare.