bluuebunny/crossref_metadata_2025 huggingface dataset:
Invasive Fungal Disease Following Myeloablative Cord Blood Transplantation for Patients with Hematological Malignancies
Authors: Li Duihong , Jiafu Huang , Li Xiaofan , Jing Zheng , Fu Haiying , Xu Zhenshu , Chen Yuanzhong , LI Nainong
Published: November 2019

Objective: Invasive fungal disease (IFD) is a serious complication after allogeneic hematopoietic stem cell transplantation (HSCT). However, limited data is available on its clinical features after cord blood transplantation (CBT) for patients with hematological malignancies.

Method: We retrospectively reviewed 63 patients who underwent myeloablative CBT at our institution between July 2013 and June 2019 with a median follow-up of 14 months. Micafungin, Voriconzole or Posaconazole was used for IFD prophylaxis.

Results: Eight patients were identified as having an IFD, including 2 with proven IFD, 1 with probable IFD, and 5 with possible IFD. The most common prophylaxis regimen is micafungin (68.2%, 43/63). The incidence rate of IFD in the primary antifungal prophylaxis (PAP) and secondary antifungal prophylaxis (SAP) groups were 11.3% and 25% (P=0.488). The OS of day at +100, six months and +200 in the IFD and No IFD groups were 62.5% vs. 86.9% (P=0.1), 50 % vs.78.2% (P=0.07), and 50% vs.75.9% (P=0.094), respectively. The 3-year probabilities of overall survival (OS) in the IFD and No IFD groups were 50% and 60.5%, respectively (p=0.316). The incidence rate of Non-relapse Mortality in the IFD and No IFD groups were 42.9% and 30%, respectively (p=0.319).

Conclusion: Antifungal prophylaxis could help to reduce the incidence of IFD after CBT for patients with hematological malignancies.

Figure

Disclosures

No relevant conflicts of interest to declare.
Clinical Utility of Flow Cytometry Immunophenotyping in Acute Myeloid Leukemia
Authors: Oliveira Gustavo Henrique de Medeiros , Júnior Lenilton Silva DA Silva , Silva Antonia Eduarda Martins Oliveira Elói , Lima João Pedro Andrade , Soares Victor lima , Freitas Rodrigo Villar , Oliveira Taissa Moura , Silva Dany Geraldo Kramer Cavalcanti , Sales Valéria SF , Paiva Hugo Diogenes De Oliveira , Araujo Rafael Santiago , Paiva Aldair Sousa , Cavalcanti Geraldo Barroso , Silva Alessandra Suelen Jardim
Published: November 2019

INTRUDUCTION: Acute myeloid leukemia (AML) is a heterogeneous group of clonal disease of hematopoietic tissue, characterized by proliferation of abnormal myeloid progenitor cells, suppressing the normal hematopoietic activity and constituting a great diagnostic challenge. With the advent of immunophenotyping by flow cytometry, the diagnosis of these neoplasms became more faithful, facilitating the treatment and follow-up of the patients. METHODS: In this study a flow cytometric immunophenotyping study was performed in bone marrow aspirate and / or peripheral blood samples from 38 patients with AML, using a panel of monoclonal antibodies specific for acute leukemias, also investigating clinical, laboratory and demographic data of these patients. RESULTS AND DISCUSSION: Of the 38 subjects studied, 23 were male and 15 females. In relation to the age, we found a higher number of cases in adult patients. According to clinical data, splenomegaly and hepatomegaly were present in most cases. The immunophenotyping demonstrated a characteristic profile of AML with expression of CD13 and CD33 in all cases and CD34 and CD117 in most cases. The CD14 was reactive in monocytic leukemia (M4 and M5-AML). Were observed negativity for lymphoid antigens CD19, CD10 and CD3 and positivity to CD7, CD56 and CD2, present in 10, 5 and one cases respectively. Regarding cytomorphology, there was a direct correlation with FAB classification, with the prevalence of type myeloid-monocytic (AML-M4). CONCLUSION: These data demonstrate the importance of immunophenotyping in the differential diagnosis of AML and monitoring of these neoplasms.

Disclosures

No relevant conflicts of interest to declare.
One-Year Outcomes with Once or Twice Daily Enoxaparin
Authors: Klil-Drori Adi J. , Nazha Sara , Gharib Marlene , Perreault Sylvie , Tagalakis Vicky
Published: November 2019

Background: Enoxaparin given once daily (QD) for thrombotic disorders is less burdensome than twice-daily (BID) dosing. However, long-term outcomes when enoxaparin given as QD monotherapy are unknown.

Methods: We did a population-based cohort study. New users of enoxaparin alone (2005-2014) were identified in the linked healthcare databases of Quebec, Canada, and followed up for up to one year. The number of dispensed syringes divided by prescription length determined QD or BID enoxaparin by intention to treat. Cumulative rates of major bleeding and re-treatment with anticoagulants at one year were compared between enoxaparin groups. Re-treatment was initiation of anticoagulation after at least 30 days of no dispensed anticoagulants. The duration of enoxaparin monotherapy was the sum of prescriptions until discontinuation.

Results: The cohort included 504 patients; QD and BID enoxaparin users were 445 and 59, respectively. Mean (SD) age was 78.0 (6.6) years, 43.8% were males, and 61.9% had cancer. At 12 months, major bleeding occurred in 21 (4.7%) and 4 (6.8%) among QD and BID enoxaparin users, respectively (Figure 1A, P = 0.49). Re-treatment with anticoagulants occurred in 37 (9.6%) and 5 (9.4%) of QD and BID users, respectively (Figure 1B, P = 0.98). The duration of enoxaparin monotherapy was on average 13.9 (95% CI, 4.4-23.4, P = 0.005) days longer with QD vs BID use.

Conclusions: Monotherapy with QD enoxaparin was common and longer than BID enoxaparin with no apparent differences in bleeding or re-treatment with anticoagulants.

Figure 1

Disclosures

Klil-Drori: Sanofi Canada: Research Funding. Nazha:Sanofi Canada: Employment. Gharib:Sanofi Canada: Employment. Perreault:Sanofi Canada: Research Funding. Tagalakis:Servier: Other: participated on ad boards; Bayer: Other: participated on ad boards; Pfizer: Other: participated on ad boards; Sanofi Aventis: Other: investigator initiated grant;participated on ad boards; BMS-Pfizer: Other: participated on ad boards.
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.
The Study of Proliferation Relative Long Non-Coding RNA in CD59- Cell from Paroxysmalnocturnal Hemoglobinuria Patients
Authors: Fu Rong , Wang Honglei , Liu Hui , Shao Zonghong , Li Liyan
Published: November 2019

Background: Paroxysmal nocturnal hemoglobinuria (PNH) is a rare clonogenic disease of hematopoietic stem cells. LncRNA has a wide range of biological functions, including regulation of gene expression, cell differentiation, cell proliferation and substance metabolism.LncRNA maybe contribute to the proliferation of PNH clone.

Methods:

PNH clone (CD59-cells) and (CD59+cells) from PNH patients were sorted and analyzed by RNA sequencing in 5 PNH patients. The results were analyzed by KEGG, we focus on the proliferation relative pathway-NF-kB pathway. The mRNA(TAB2, TLR4, LYN,CFLAR, TNFAIP3, PTGS2, TRIM25, CXCL8) which FPKM>10 and over 3 pathients were chosen to search out the upstream regulation LncRNA. MALAT1 and LINC01002 were screened by Co-expression. Then the expression of MALAT1 and LINC01002 in 30 PNH patients were detected by qRT-PCR to vertify the LncRNA sequencing results.

Results: Transcription analysis revealed that 742 upregulation LncRNA and 3276 upregulation mRNA were identified in CD59- cells. The highly expressed NF-kB pathway mRNA (TAB2, TLR4, LYN,CFLAR, TNFAIP3, PTGS2, TRIM25, CXCL8) were analysed by LncRNA co-expression, after that MALAT1and LINC01002 were concerned with the 8 mRNA. The results of PNH primary cells (CD59-cells) showed that the level of MALAT1and LINC01002 expression was significantly higher than that of the CD59+ cell in 30 PNH patients (p<0.05).

Conclusion: MALAT1and LINC01002 seems to be involved in the proliferation of PNH clones. Its mechanism of action in patients with PNH needs further study.

Disclosures

No relevant conflicts of interest to declare.
Faster Diagnosis of TTP Using New Technoscreen Adamts-13 Activity Assay
Authors: Braf Annica , von Horn Henrik , Bark Niklas , Norberg Eva-Marie , Truedsson Asa , Binder Nikolaus B , Antovic Jovan P
Published: November 2019

Background

The thrombocytopenic purpura (TTP) is a life-threatening condition characterized by microvascular thrombosis where it is of great importance to shorten the time to the treatment. TTP can be caused by congenital or acquired deficiency of ADAMTS13 (an enzyme that cleaves the von Willebrand factor). Deficiency leads to the accumulation of ultra-large von Willebrand multimers, which causes platelet aggregation and the formation of microthrombi. TTP laboratory findings include thrombocytopenia, hemolytic anemia, the presence of schistocytes and a markedly reduced activity of ADAMTS13. ADAMTS13 activity can be analyzed using an ELISA method. We have evaluated whether a new semi-quantitative method, TECHNOSCREEN from Technoclone, can be used as a screening method to shorten the time for patient diagnosis.

Method

We analyzed 43 unidentified samples with known ADAMTS 13 activity, which consisted of 20 samples with an ADAMTS13 activity <10%. The new screening method (TECHNOSCREEN ADAMTS13 Activity) was compared to the existing ELISA method (TECHNOZYM ADAMTS13 Activity ELISA). Both methods were from Technoclone, Vienna, Austria. The methods are based on incubating patient plasma with von Willebrand fragments and specific antibodies to cleavage fragment. Color intensity correlates with the amount of cleavage and thus ADAMTS13 activity of the sample. In the screening method, color development takes place in a flow through test device and is semi-quantitatively graded by visual comparison against a color chart graded as 0.0, 0.1, 0.4 and 0.8 IU /mL.

Results and conclusion

The results showed good agreement between the existing ELISA and the newTECHNOSCREEN method. Sensitivity was 100% and specificity 71% if the ELISA is used as a gold standard. In May 2019, the TECHNOSCREEN method was introduced as a first-line analysis. This has resulted in faster patient diagnosis as well as saving laboratory staff time. The TECHNOSCREEN assay takes about 1 hour to perform compared to about 5 hours for the ELISA. To exactly quantify ADAMTS13 activity or detect the presence of antibodies, the ELISAs are performed approximately once a week.

Disclosures

Binder: Technoclone: Employment. Antovic:Shire: Other: Grants and Personal Fees; Roche: Other: Personal Fees; Stago: Other: Personal Fees; Werfen: Other: Personal Fees; Siemens: Other: Personal Fees; Sobi: Other: Personal Fees; Novo Nordisk: Membership on an entity's Board of Directors or advisory committees, Other: Personal fees; CSL Behring: Other: Personal Fees.
Prevalence of Bronchiolitis Obliterans Syndrome (BOS) Following Allogeneic Hematopoietic Stem Cell Transplant (alloHSCT) in the USA, Europe and Japan
Authors: Hofstetter Emilie , Henig Noreen Roth , Kappeler Dominik
Published: November 2019

Background:

Bronchiolitis obliterans syndrome (BOS), also known as respiratory chronic Graft versus Host Disease (cGvHD) is an obstructive airway disease of the lungs mainly associated with alloHSCT or lung transplantation. BOS is characterized by T-cell mediated inflammation and fibrosis of bronchiolar walls that reduces the diameter of the bronchioles resulting in progressive and irreversible airflow obstruction.

BOS is a well described complication following lung transplant with a 5-year prevalence rate of 50%. It can also be a complication following alloHSCT with similar histopathology and clinical symptoms. Currently, there are no approved treatments for BOS. The aim of this study is to describe the prevalence of BOS following alloHSCT.

A review and analysis of the following data sources was performed: AlloHSCT activity reports from The European Society for Blood and Marrow Transplant 2016 the Center for International Blood and Marrow Transplant Research 2016, and the Japanese Data Center Hematopoeitic Stem Cell Transplantation 2015 in order to identify the number of procedures performed. Further a PubMED literature search was also conducted and identified publications with > 100 patients using the key terms "bronchiolitis obliterans and haematopoietic stem cell", and "bronchiolitis obliterans after stem cell transplant" from 2011 to 2017. Case reports and redundant publications were excluded from the analysis.

Results

Approximately 30,000 alloHSCTs were performed each year in the USA (8766 in 2016), Europe (17,617 in 2016) and Japan (3700 in 2015). Based on seven eligible studies published between 2011 and 2016, the reported prevalence of BOS in patients who underwent alloHSCT ranged from 3.4% to 10%. Median time to diagnosis of BOS following alloHSCT was 273 to 547 days post-transplant. Those with routine Forced Expiratory Volume at 1 second (FEV1) monitoring had a higher likelihood of being diagnosed with BOS and a shorter time to diagnosis.

Conclusion

Based on estimated prevalence, up to 3000 patients following alloHSCT per year could be diagnosed with BOS. Routine monitoring may positively impact early and shorten time to diagnosis and allow treating such a population.

Figure

Disclosures

Hofstetter: Breath Therapeutics GmbH: Consultancy. Henig:Breath Therapeutics Inc.: Employment. Kappeler:Breath Therapeutics GmbH: Consultancy.
Free Fatty Acid Uptake By Hematopoietic Stem and Progenitor Cells Drives Immune Cell Expansion in Response to Salmonella Typhimurium infection
Authors: Mistry Jayna J , Moore Jamie A , Hellmich Charlotte , Marlein Christopher R , Beraza Naiara , Di Palma Federica , Bowles Kristian M , Rushworth Stuart A
Published: November 2019

Haematopoietic stem and progenitor cells (HSPC) are tightly regulated in a balance between self-renewal and differentiation and undergo rapid expansion in response to infection. In the context of acute bacterial infection, the hematopoietic system needs energy to drive the immune response necessary for host survival. In an adult human, bone marrow adipose tissue accounts for up to 70% of bone marrow and we have previously shown that free fatty acids are transferred from adipocytes to AML blasts providing the AML blasts more energy for proliferation and survival (Shafat, Blood 2017). Therefore, we hypothesis that during an acute bacterial infection, the HSPC population uptakes free fatty acids (FFA) to support and sustain the rapid response to bacterial infection. To determine if HSPC acquired FFA we infected C57BL/6 with Salmonella Typhimurium (S.typhimurium) or treated C57BL/6 with lipopolysaccharide (LPS) for 2 and 16 hours. Using both models we show that the HSPC populations (HSC (haematopoietic stem cell), MPP (multipotent progenitor) and LSK (LN-, CD117+, Sca1+)) all accumulate FFA as measured by staining with neutral lipid-specific BODIPY 493/503 dye. To determine if this was uptake or accumulation of fatty acid within the HSPC of animals infected with S.typhimurium we used BODIPY®-dodecanoic acid fluorescent fatty acid to trace FFA uptake. HSPC populations from animals treated with LPS had a higher uptake of BODIPY®-dodecanoic acid when compared to the control treated mice. To assess metabolic changes within HSC population in response to infection, cells were sorted (LN-, CD117+, Sca1+, CD48-, CD150+ and CD34+) and analysed for gene expression changes. In HSC, increases were observed in several metabolic genes including FABP4, CD36 and CPT1 in response to S.typhimurium as well as LPS at 16 hours post stimulation. Moreover, following cell sorting, the LSK (LN-, CD117+, Sca1+) population were assessed for metabolic activity for different substrates. The LSK population from LPS treated animals had an increased dependency on beta oxidation when compared to the control. Moreover, etoximir (a beta-oxidation inhibitor) inhibited LSK basal metabolism and reduced LSK expansion (as measured by colony forming cell assay and Ki-67 staining) in animals treated with LPS. Our data indicates that infection drives uptake of FFA in different HSPC populations. The FFA are metabolised by beta oxidation which provides the energy for the expansion of leukocytes.

Disclosures

Bowles: Abbvie: Research Funding; Janssen: Research Funding. Rushworth:Abbvie: Research Funding; Janssen: Research Funding.
Novel Immune Checkpoint Deficiency and Susceptibility to EBV-Associated Lymphoma
Authors: Somekh Ido , Magg Thomas , Duque Alejandro Gallón , Stauber Tali , Lev Atar , Unal Ekrem , Simon Amos J. , Rohlfs Meino , Conca Raffaele , Tatematsu Megumi , Hauck Fabian , Somech Raz , Klein Christoph
Published: November 2019

Background: Understanding genetic predisposition to cancer is of paramount importance to tailor therapeutic strategies. Several defects in immune checkpoint regulators have been discovered in patients with EBV-induced lymphoma (e.g. CD27, PRKCD, RASGRP1, MAGT1, SH2D1A, ITK). Here, we describe the clinical and immune phenotype of 2 unrelated patients from consanguineous families presenting with a primary immune deficiency (PID) and EBV-associated lymphoproliferation due to biallelic mutations in CD137 (TNFRSF9/4-1BB).

Methods: One patient of Turkish origin (P1) and Palestinian origin (P2), respectively were evaluated. Genetic analysis using whole exome sequencing was conducted. Immunological and biochemical assays were performed on primary patient material.

Results: Clinical findings included recurrent sinopulmonary and herpes virus infections from childhood on. One patient suffered from auto-immunity (AIHA and auto-immune thrombocytopenia). Abnormal immunoglobulin levels were documented (IgG 413-1670, IgM 105-714, IgA 49-68 mg/dL). Both patients developed EBV-associated lymphoproliferative disorders: P1 developed Burkitt lymphoma and was treated according to NHL-BFM2000 regimen in combination with rituximab. He is currently in remission. P2 had monoclonal EBV-positive lymphoproliferation and was successfully treated with immunosuppressive therapy (e.g. cellcept, glucocorticoids). To shed light on the underlying genetic etiology, we performed whole exome sequencing. A large homozygous deletion in CD137 (c.1_545+1716del) was identified for P1, while P2 harbored a homozygous missense mutation (c.C452T, p.Thr151Met). Impaired anti-CD3 T cell lymphocyte activation and proliferation were observed, amenable to correction upon addition of anti-CD28 monoclonal antibodies. In an attempt to provide definitive proof that the CD137 gene variant causes the activation defect of T-cells, we designed a genetic rescue experiment in patient T cells. Upon retrovirus-mediated recombinant expression of WT CD137, proliferation and activation defects in T cells were restored.

Conclusions: In sum, we here show that a genetic defect in the immune checkpoint molecule CD137 causes a new primary immunodeficiency disorder with susceptibility to EBV-induced lymphomagenesis.

Disclosures

No relevant conflicts of interest to declare.
Improved Outcomes of All-Trans-Retinoic Acid and Arsenic Trioxide Plus Idarubicin As a Frontline Treatment in Adult Patients with Acute Promyelocytic Leukemia
Authors: Yang Shiwei , Zhu Zunmin , Zhu Xiaojian , Zhang Yicheng
Published: November 2019

Objective To analyze the outcomes of all-trans-retinoic acid (ATRA) and IV arsenic trioxide (ATO) plus idarubicin (IDA) as a frontline induction therapy in 118 acute promyelocytic leukemia (APL) patients with high risk (HR) and standard risk (SR) from January 2008 to December 2017. Methods The medical records of 118 APL patients (HR, n=45; SR, n=73) who received the frontline triple combination regimen at Henan Provincial People's Hospital and Tongji Hospital were retrospectively reviewed. Consolidation therapy comprised 6 cycles of ATO and ATRA plus IDA, and IDA was administered in 1~4 cycles of consolidation therapy based on the comparable clinical efficacy compared with 6 cycles and fewer side effects to myocardium without subsequent maintenance therapy. Results Of 118 patients, there were 3 (2.5%) early deaths and 115 (97.5%) hematologic complete remissions (hCR), and 102 (88.7%) of 115 patients achieved molecular complete remissions (mCR) following induction therapy, and all patients were PCR-negative for PML-RARa after the first cycle of consolidation therapy. The 5-year overall survival (OS) and event-free survival (EFS) were 93.0±2.9% and 92.4±3.0%, respectively. Early death, hCR, mCR and toxicities were comparable between the HR and SR groups. The cumulative incidence of relapse (CIR) in the HR group was 4.7% (n=2), and the CIR in the SR group was 0. The OS and EFS of the SR and HR groups were comparable (92.3±4.5 vs. 92.8±4.0%, X2=0.263, P=0.608; 92.3±4.5 vs. 91.1±4.2%, X2=0.917, P=0.338). The total dosage of IDA was approximately 181~258 mg, and no patient experienced cardiotoxicity. OS and EFS were not influenced by fms-related tyrosine kinase 3 internal tandem duplication (FLT3-ITD) mutation status (P=0.405, 0.528, respectively). Conclusion The triple combination of ATRA and ATO plus IDA as both an induction and consolidation therapy for the HR and SR groups attained excellent outcomes, and this regimen was effective, safe and easy, without maintenance therapy. The triple combination treatment might be a preferred frontline therapy for APL patients, especially for HR patients or patients with the APL FLT3-ITD mutation.

Disclosures

No relevant conflicts of interest to declare.
This Is a Title in Title Case: A Combination of Low-Dose Decitabine and Chidamide Resulted in Synergistic Effects on the Proliferation and Apoptosis of Human Myeloid Leukemia Cell Lines
Authors: Xu Fangfang , Guo Honggang , Shi Mingyue , Liu Siwei , Wei Min , Sun Kai , Chen Yuqing
Published: November 2019

Introduction: Myeloid leukemia is a malignant disease caused by both genetic and epigenetic changes. Aberrant DNA methylation and histone modifications are prevalent in cancers including leukemia and the process of epigenetic modifications is reversible, which makes them potential treatment targets using specific inhibitors. In this study, we sought to determine the antileukemic effects of low-dose decitabine (one of the most widely used DNA methyltransferase inhibitor) combined with chidamide (a novel orally active histone deacetylase inhibitor) on myeloid leukemia cells by detecting cell proliferation, cell cycle distribution and cell apoptosis to provide a promising regimen for clinical application.

Methods: We conducted CCK-8 assay to determine the effect of low-dose decitabine combinated with chidamide on cell viability, cell cycle distribution analysis by use of flow cytometry accompanied with RT-qPCR and western blotting analysis to reveal its inhibitory mechanism of cell growth, and cell apoptotic analysis also by use of flow cytometry accompanied with RT-qPCR, western blotting analysis, intracellular ROS production and mitochondrial transmembrane potential assay to determine the apoptotic mechanism of combination of low-dose decitabine and chidamide in K562 and THP-1 cells.

Result: Low-dose decitabine and chidamide alone played an inhibitory role in the proliferation of K562 and THP-1 cells in a dose- and time-dependent manner, and the combination of the two enhanced this effect, accompanied by induction of p21 expression and cell cycle arrest at G0/G1 phase. Chidamide instead of decitabine had pronounced effects on the histone H3 acetylation levels of leukemia cells. Meanwhile, combination of the two significantly induced cell apoptosis via mitochondrial transmembrane potential loss, resulting in upregulation of the cell apoptosis-related gene Bax and the protein cleaved PARP-1 and downregulation of antiapoptosis gene Bcl-2, and proteins, including pro-caspase 9, pro-caspase 3 and PARP-1.

Conclusion: Our results provided evidence for the antileukemia effects of low-dose DNA methyltransferase inhibitor decitabine combinated with histone deacetylase inhibitor chidamide in preclinical myeloid leukemia models and suggested that combination of the two showed therapeutic potential for myeloid leukemia treatment.

Disclosures

No relevant conflicts of interest to declare.