bluuebunny/crossref_metadata_2025 huggingface dataset:
Resistance to Nutrient Deprivation Via Metabolic Re-Programming As Mechanism of 'Metastasis' Promotion in Multiple Myeloma
Authors: Savvidou Ioanna , Chandrasekara Sahan , Chenn Maoshan , Khong Tiffany , Spencer Andrew
Published: October 2019

Aim: Multiple Myeloma (MM) is a disease of the intra-medullary (IM) compartment but with progression myeloma cells can survive in nutrient deprived extramedullary (EM) sites. Autophagy mediates the degradation and re-cycling of intra-cellular proteins and may promote tumor resistance to metabolic stressors, including nutrient deprivation. Using the MM cell lines TK1 and TK2, contemporaneously propagated from bone marrow (IM) and peripheral blood (EM), respectively, of a MM patient we investigated the role of autophagy in resistance to glutamine (Gln) deprivation.

Methods & Results:TK2 but not TK1 exhibited up-regulated autophagy under both basal and Gln deprived conditions with higher LC3BII/I turnover on immunoblotting and autophagic vacuole formation on electron microscopy following chloroquine (CQ) exposure. Under conditions of Gln deprivation, TK2 was able to proliferate until day 14, whereas TK1 stopped proliferating at day 3. This proliferative advantage under Gln deprivation was abrogated by autophagy inhibitors (CQ or 3-MA) as determined by both viable cell enumeration (p=0.008 at day 7, inhibitor vs no inhibitor) and Ki67 expression (p=0.0017) only in TK2. Modulation of Gln concentration demonstrated an inverse correlation with the viability of TK1 but not TK2 - TK1 cell death increased from 36% at Gln 8mM to 56% at Gln 0mM, irrespective of autophagy inhibition. In contrast, while Gln deprivation had no significant effect on the viability of TK2, under Gln deprivation CQ exposure induced a 2-fold increase cell death measured by annexin V/PI positivity and 2.5-fold increase of cPARP expression (p=0.0045).

RNAseq and GSEA analysis showed enrichment of the Krige Amino Acid Deprivation Response (AADR) pathway (ES=0.083) for TK1 with up-regulated expression of Gln transporters (SLC7A5, SLC1A5, SLC38A5 and SLC7A11). In contrast, Glutamine Synthetase (GS) expression, which is responsible for Gln synthesis, was up-regulated in TK2 cells. Interestingly, immunoblotting analysis revealed that Gln deprivation was able to induce the expression of GS while further inducing autophagic flux (2.66 fold increase of LC3II expression at Gln=0mM) only in TK2, whereas it inhibited mTOR activity in both cell lines. Additionally, GS chemical inhibition under Gln deprivation induced cell death in TK2 but not TK1 (32.56% vs 2.5% increase, respectively) demonstrating that Gln synthesis enables TK2 to overcome Gln deprivation.

Conclusion. Metabolic reprogramming mitigating against nutrient deprivation may promote a more 'metastatic' phenotype in advanced MM.

Disclosures

Spencer: Celgene: Honoraria, Research Funding, Speakers Bureau; Janssen-Cilag: Honoraria, Research Funding, Speakers Bureau; Amgen: Honoraria, Research Funding; BMS: Research Funding; Takeda: Honoraria, Research Funding, Speakers Bureau; STA: Honoraria.
The Role of Nestin+ Mesenchymal Stem Cells (MSCs) in Bone Marrow Chronic Graft-Versus-Host Disease
Authors: Zhang Haiyan , Yang Kaibo , Chen Yanqiu , Jin Hua , Liu Qifa
Published: October 2019

Chronic graft-versus-host disease (cGVHD) is a major cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). cGVHD is a syndrome of variable clinical features resembling autoimmune, involving not only the epithelial target tissues (skin, lung, gastrointestinal tract and liver) but also any other organ system (musculoskeletal, joint, ocular and genital tissues). Bone marrow (BM) suppression without infection is often observed in patients undergoing allo-HSCT as GVHD symptoms appear, suggesting that BM could be a target of GVHD (Yusuke S. et. al. Blood 2010). However, the BM cGVHD is rarely reported and the mechanism for BM cGVHD is still unclear. Nestin+ mesenchymal stem cells (MSCs) are capable of differentiating into cells of multiple cell lineages and involved in the formation of fibrotic lesions (Miaohua M. et. al. Cell Mol Life Sci 2016). To determine whether Nestin+ MSCs play an important role in the development of BM cGVHD. We used two murine models of cGVHD, including MHC-matched BALB/c (H-2d) recipients given B10D2 (H-2d) grafts, and MHC-mismatched BALB/c (H-2d) recipients given C57BL/6 (H-2b) grafts. We observed that BM displayed a significant increase in collagen and reticulin fibers deposition around blood vessels in cGVHD group compared to non-cGVHD group. The decreased absolute numbers of monocytes and neutrophils in the BM indicated the broad BM suppression in the cGVHD group. We further demonstrated that Nestin+MSCs located in perivascular BM niche underwent tremendous expansion in cGVHD group (p<0.001). Nestin+ MSCs in cGVHD group acquired expression of alpha smooth muscle actin (a-SMA), indicating myofibroblast differentiation. Moreover, transforming growth factor- beta1 (TGF-β1) concentrations also increased in cGVHD BM microenvironment (p<0.0001). High concentrations of TGF-β1 induced formation of Nestin+ MSC clusters and more transition from Nestin+ MSCs to a-SMA+ cells (p<0.05). Besides, expanded Nestin+ MSCs were surrounded by many macrophages and expressed high level of TGF-β1 receptor (p<0.001 and p<0.005). Taken together, these results indicate that Nestin+ MSCs participate BM cGVHD development through transition to a-SMA+ cells induced by high TGF-β1 in cGVHD microenvironment.

Disclosures

Zhang: National Natural Science Foundation of China (No. 81600141, No. 81770190) and Natural Science Foundation of Guangdong Province (No. 2016A030310390): Research Funding. Yang:National Natural Science Foundation of China (No. 81600141, No. 81770190) and Natural Science Foundation of Guangdong Province (No. 2016A030310390): Research Funding. Chen:National Natural Science Foundation of China (No. 81600141, No. 81770190) and Natural Science Foundation of Guangdong Province (No. 2016A030310390): Research Funding.
Immune-Mediated Hematologic Complications of Checkpoint Inhibitors: A Review of the Literature
Authors: Sallam Omar , Riaz Irbaz Bin , Go Ronald S.
Published: October 2019

Background:

The incidence and types of hematologic complications from immune check point inhibitors are not well known. We conducted this review to describe immune-mediated hematologic complications reported in clinical trials, case series, and case reports.

Methods:

A pre-defined comprehensive search strategy was used to identify case reports, case series, and clinical trials using PubMed. Any study that reported hematologic complications was included. Data were extracted for demographic characteristics and occurrence of immune-mediated hematologic complications. We pooled the data to calculate the frequency of immune-mediated hematologic adverse effects.

Results:

A total of 689 of studies were retrieved using the search criteria and 75 were included in the analysis (31 case reports and case series and 44 clinical trials). There were 44 patients reported having immune-mediated hematologic complications, 4 of them in clinical trials. The complications included aplastic anemia, autoimmune hemolytic anemia, cryoglobulinemia, graft versus host disease, hemophilia A (acquired), immune neutropenia, immune thrombocytopenia, macrophage activation syndrome, myelodysplastic syndrome, pure red cell aplasia, and thrombotic thrombocytopenic purpura. However, the overall rates were very low, ranging from 1-2.2% in clinical trials. Immune thrombocytopenia was the most common (29.5%), followed by autoimmune hemolytic anemia (15.9%), and immune neutropenia (13.6%). Immune-mediated hematologic complications were reported in all classes of checkpoint inhibitors including anti-programmed cell death protein 1 (nivolumab and pembrolizumab), anti-programmed death ligand 1 (avelumab and durvalumab), and anti-cytotoxic T-lymphocyte-associated protein 4 inhibitors (ipilimumab and tremelimumab). Among patients reported in case reports and case series, the median age was 57 years (range, 29-85) and most were males (52.9%). The majority of the complications occurred in patients treated with ipilimumab (38.8%), nivolumab (27.7%) and pembrolizumab (16.0%). The onset was usually within the first week of receiving the first dose but could occur up to 17 months after drug initiation. Indefinite discontinuation of the immunotherapy was the mainstay of treatment resulting in resolution of complications in the majority (74.5%) of the patients. Two patients were re-challenged with the same checkpoint inhibitor and one experienced a relapse of immune cytopenia (autoimmune hemolytic anemia).

Conclusion:

Immune-mediated hematologic complications associated with checkpoint inhibitors are rare. They are usually reversible after discontinuation of such treatment. Relapses may occur with re-challenge.

Disclosures

No relevant conflicts of interest to declare.
Neutrophil KLF2 Regulates Arterial and Venous Thrombosis
Authors: Nayak Lalitha V. , Lapping Stephanie , Maiseyeu Andrei , Schmaier Alvin H. , Jain Mukesh
Published: October 2019

Experimental, clinical and pathological studies support an important link between inflammation and thrombosis. Although accumulating evidence suggests that cells of the innate immune system contribute to the thrombotic process, the identity of nodal molecular determinants operative in immune cells remains poorly understood. Our previous work in mice bearing global deletion of the transcription factor KLF2 identifiedthis factor as a critical mediator of thrombosis). Here, using cell-specific KLF2 deleted murine models (endothelial, platelet, and myeloid- deleted KLF2) we identify myeloid KLF2 as the major determinant of thrombosis. Complete blood counts and coagulation assays in myeloid KLF2 deleted mice (MY-K2-KO) were similar to those in control LysM cre mice. Using two models of vascular thrombosis (carotid artery thrombosis assay, Rose Bengal model and complete inferior vena cava ligation model of venous thrombosis) we observed a robust prothrombotic phenotype in the MY-K2-KO mice as compared to control Cre mice. As LysM Cre deletes in both neutrophils and monocyte/macrophages, we next sought to determine whether one subset was dominantly responsible for the observed phenotype. Adoptive transfer of KLF2-KO neutrophils in control mice conferred both the arterial and venous pro-thrombotic phenotype. Conversely, neutrophil depletion with Ly6G antibody (>90% depletion achieved) reversed the prothrombotic phenotype in MY-K2-KO mice. By contrast, depletion of monocytes/macrophages using clodronate liposome injection did not affect arterial or venous thrombosis. Next, we evaluated the effect of KLF2 deficiency on critical neutrophil functions that may contribute to the pro-thrombotic phenotype. As compared to controls, MY-K2-KO neutrophils demonstrated more pronounced recruitment to the endothelium. In microflow chamber assays, MY-K2-KO neutrophils showed decreased rolling and increased P-selectin induced adhesion. The importance of this was affirmed by the observation that adoptive transfer of MY-K2-KO neutrophils incubated with PSGL-1 antibody was unable to generate a prothrombotic phenotype in control mice. In addition, MY-K2-KO neutrophils demonstrated enhanced neutrophil extracellular traps generation associated with increased myeloperoxidase and neutrophil elastase expression. And finally, MY-K2-KO neutrophils (but not monocytes) demonstrated increased tissue factor expression and activity. Collectively, these studiesidentify neutrophil KLF2 as a critical determinant of both arterial and venous thrombosis and suggest that modulation of neutrophil KLF2 may be a fruitful approach to the management of thrombosis.

Disclosures

Schmaier: Alnylam: Research Funding; Biomotiv: Consultancy; Cleveland Clinic Foundation: Research Funding; Temple University: Patents & Royalties; Enzyme Research Laboratories: Honoraria; Shire: Consultancy, Honoraria, Research Funding.
Sequential Infusion of Anti-CD22 and Anti-CD19 Chimeric Antigen Receptor T Cells Following Autologous HSCT in Patients with B-NHL
Authors: Cao Yang , Wang Na , Wang Gaoxiang , Xiao Yi , Huang Liang , Li Chunrui , Xiao Min , Zhang Tongcun , Zhang Yicheng , Zhou Jianfeng
Published: October 2019

Salvage chemotherapy followed by autologous hematopoietic stem cell transplantation (HSCT) is the standard treatment for the patients with chemotherapy sensitive relapsed or refractory B-NHL. However, more than half of patients will ultimately relapse. To improve long-term remission rate, we administered adoptive T-cell immunotherapy after HSCT. Based on the findings that CD19/CD22 dual antigen targeting have been demonstrated to be a promising approach for overcoming antigen escape relapse, we conducted an open-label, single-center and single-arm pilot study of sequential infusion of anti-CD22 and anti-CD19 CAR-T cells following autologous HSCT. We aimed to evaluate its safety and efficacy in patients with relapsed, refractory and high-risk B cell lymphoma. This trial is registered with ChiCTR, number ChiCTR-OPN-16009847.

Between December 2016 and March 2018, 20 patients were enrolled in this clinical trial, with a median age of 33 years (range, 24-61 years). Of these 20 treated patients, 2 had PMBCL, 4 had tFL,14 had DLBCL, 70.0% (14/20) had received at least three previous therapies. 50.0% patients (8/16) harbor a TP53 mutation. At baseline assessment (prior to conditioning), 3 participants had a CR, 8 had a PR, 8 had a PD, 1 had a SD. All patients received bis-chloroethyl nitrosourea, etoposide, Ara-C, and melphalan, CD22 and CD19 CAR T cells were infused 2 to 7 days after stem cell infusion. The median anti-CD22 and anti-CD19 CAR-T cells dose per kilogram of body weight were 4.0× 106 (range, 1.0-10.0×106) and 4.2× 106 (range, 1.8-10.0×106) respectively. Normal HSCT-associated neutrophil engraftment within 20 days in all 20 patients was observed. 11/20 (50.5%) patients experienced cytokine release syndrome and no patient had severe CRS (grade 3-4). Neurologic events occurred in 2 (10.0%) patients. At 3 months, there was a 90.0% remission rate (95% CI ,68.3-98.8) with 85.0 % complete remissions (CRs) and 5.0% partial remissions (PRs) accessed by PET-CT. The duration of responses currently ranges from 3.7 months to19.1 months. All patients (17/20) who achieved CRs are ongoing, and the current median duration of all CRs is 13.2 months.

Our results indicated that patients with relapsed, refractory and high-risk B cell lymphoma who received sequential infusion of anti-CD22 and anti-CD19 CAR-T cell therapy following HSCT had high rates of durable remission, with safety profile. Anti-CD22 and Anti-CD19 CAR-T cell therapy following HSCT may become a useful treatment approach for relapsed , refractory B cell lymphoma and high-risk B cell lymphoma.

Disclosures

No relevant conflicts of interest to declare.
Compound Application of Thalidomide and Recombinant Human Interferon-α-1b, Interleukin-2 in Different Disease Status of Acute Myeloid Leukemia a Multi-Center Prospective Study of a New Combination of Old Drugs
Authors: Chen Lin , Mi Ruihua , Wei Xudong , Ying Qingsong , Yuan Fangfang , Liang Lijie , Li Mengjuan , Liu Delong , Song Yongping
Published: October 2019

Objectives: The aim of this study was to investigate the therapeutic effects of combinational application of thalidomide and recombinant human interferon-α-1b, interleukin-2 (the ITI scheme) in treating acute myeloid leukemia (AML) in different settings.

Methods: AML patients with different disease states were categorized into three groups and received treatments following the ITI scheme. Group A consisted of relapsed or refractory AML patients (R/R-AML) and primary AML patients who were unable to receive chemotherapy. Group B included patients with morphologically complete remission (CR) and consistently positive minimal residual disease (MRD). Group C is composed of AML patients with initial complete remission and negative MRD. The CR rate, partial remission rate, MRD status, quality of life, and long-term survival were observed accordingly. ITI scheme is administrated as: IFNα-1b 60ug/d qod ih, IL-2 1001 million units/d qod ih, Thalidomide 200mg qn po. the Compound Salvia Tablets were recommended orally taken in order to prevent deep venous thrombosis.

Results: a. With a total response rate of 30% in 60 AML patients of Group A, 4 CR, 6 CRi and 8 PR, who maintained good quality of life with no transfusion with red blood cells or platelet components any more (Table1).The MRD of 13 cases (72.2%) turned negative among 18 patients in Group B. MRD levels turned unmeasurable in seven patients receiving a conventional dose of ITI regimen, and MRD levels significantly decreased in three patients. MRD levels turned unmeasurable after receiving a higher dose of ITI regimen in two patients, and one patients MRD level decreased. Five patients failed to this regimen and suffered a hematologic recurrence. The efficacy is positively correlated with level of MRD level before ITI administration. 81.8% (9/11) patients responded to ITI regimen in whose MRD level ≤1.0%. However, only 57.1% (4/7) responded in whose MRD≥1.0%. 3 patients responded to a extra dose of ITI after no response to a normal dose. Indicated that the efficacy of this regimen is positively correlated with the dose of the regimen.(Table 2)In group C of 88 patients, 11 patients failed to maintain MRD negative, and the relapse rate was 12.5% (Table 3and figure 1).

Conclusions: The ITI scheme can provide a new, effective, and affordable treatment option for AML patients that are intolerant to conventional chemotherapy, including R/R-AML patients, patients with CR status but MRD positive, and patients after initial CR.

Disclosures

No relevant conflicts of interest to declare.
Abnormal Coagulation Function in Chimeric Antigen Receptor T Cell Treatment of Hematologic Malignancies
Authors: Qi Kunming , Cheng Hai , Cao Jiang , Chen Wei , Qiao Jianlin , Zeng Lingyu , Li Zhenyu , Xu Kailin
Published: October 2019

Chimeric Antigen Receptor T cell( CAR-T) is an effective treatment for refractory recurrent hematologic malignancies nowdays. However, CAR T cells can potentially damage normal tissues by specifically targeting a tumor-associated antigen that is also expressed on those tissues. Cytokine release syndrome (CRS), a systemic inflammatory response caused by cytokines, is the most common type of toxicity, including fever, hyoxemia, pain and so on. In our study, we found 6 cases abnormal coagulation function which was not very common complication after CAR-T infusion. 4 cases relapsed/refractory acute B lymphoblastic leukemia(B-ALL), including 2 female 53 and 45 years old, 2 male 20 and 6 years old. 1 case multiple myeloma(MM), female 65 years old and 1 case relapsed/refractory diffuse large B-cell lymphoma (DLBCL), male 17 years old. After conditioning chemotherapy with fludarabine and cyclophosphamide, CAR-T cells were infused into patients. In these cases, female 65 years old B-ALL patient had the most severe cytokine release reactions, multiple organ function damage, and abnormal coagulation function that prothrombin time (PT) and activated partial thromboplastin time (APTT) were prolonged and reached a peak (Too long to detect) on the 14th day after CAR-T infusion, while fibrinogen (FIB) was reduce d (peak on the 14 day, 0.328g/L) and D-Dimer, fibrinogen degradation products (FDP) significantly increased (peak on the 12th day, 28.7ug/mL and more than 10mg/L respectively). After positive hemostasis and other treatments with amperenic acid, vitamin K1 and so on, the level of PT, APTT, FIB, etc were returned to normal on the 19thday. The other 5 patients went through the same process, only slightly. Patients with severe abnormal coagulation function were prone to bleed and even life-threatening, and giving active treatment, finally they all recovered. For the first time, we reported the abnormal coagulation function after CAR-T infusion in hematologic malignancies.

Disclosures

No relevant conflicts of interest to declare.
Synergic Effect of Histone Deacetylase Inhibitor Chidamide with Doxorubicin on Diffuse Large B-Cell Lymphoma with Double BCL2/MYC Expression
Authors: Fang Ying , Liu Feng , Zhao Wei-li
Published: October 2019

Introduction

Diffuse large B-cell lymphoma (DLBCL) represents the most common neoplastic disorder of B-lymphocytes. Although great progress has been made in the treatment of DLBCL, patients with double BCL2/MYC expression have been reported to be refractory to R-CHOP regimen. In our clincial trial (NCT02753647), we found that chidamide, a histone deacetylase inhibitor, plus R-CHOP elicits favorable outcome, especially for DLBCL patients with double BCL2/MYC expression. Therefore,we set out to investigate the underlying mechanism.

Methods

Core needle biopsy of tumor samples of DLBCL with double BCL2/MYC expression were transplanted into immunodecient (NOD-SCID) mice to establish patient-derived xenograft (PDX) models. Heterotopic PDX models were maintained by passaging tumor tissues from mouse to mouse. Chidamide and doxorubicin, either alone or in combination, were applied to PDX models with low passage numbers (<10) to preserve the genetic integrity of the parental tumors. The dose and administration schedule were as follows: doxorubicin 0.6mg/kg twice a week, chidamide 12.5mg/kg/day for two weeks. RNA-seq and ATAC-seq were performed to achieve an integrative transcriptomic and epigenomic anlaysis of PDX models on Day 7 and Day 14 of treatment. The study was approved by Shanghai Rui Jin Hospital Review Board and written informed consent were obtained from patients in accordance with the Declaration of Helsinki.

Results

We observed significant growth inhibition of xenografted tumors treated with chidamide combined with doxorubicin, as compared to those of the single treatment. RNA-seq analysis revealed that chidamide and doxorubicin synergistically regulate a set of genes associated with B-cell differentiation and multiple signaling transduction pathways involved regulation of BCL2 and MYC, including the IL6-JAK-STAT and PI3K-AKT-MTOR pathways. Furthermore, accessible chroamtin region profiling by ATAC-seq indicated that chidamide and doxorubin co-repress distal cis-regulatory elements (i.e. enhancers) associated with lymphocyte development.

Conclusions

As a histone deaceltylase inhibitor, chidamide is able to synergize with doxorubicin to inhibit the growth of DLBCL expressing high levels of MYC and BCL2 in PDX models. This is, in part, due to remodeling of the landscape of accessible chromatins in lymphoma cells. Our study provides insight into how epigenetic therapeutics can sensitize DLBCL with double BCL2/MYC expression to conventional chemotherapy.

Disclosures

No relevant conflicts of interest to declare.
Antiplatelet Therapy for Patients with Hemophilia after Percutaneous Coronary Intervention
Authors: Xie Peter , Malik Devin , Kuriakose Philip
Published: October 2019

Background:

The incidence of coronary artery disease is increasing in hemophilia patients as their life expectancy improves. However, there are limited evidence-based studies available, and most current guidelines for antiplatelet therapy in patients with hemophilia are largely based on expert opinions. The assessment of benefit and risk of using antiplatelet therapy in these patients remains a clinical challenge.

Aim:

This study reports our institution's clinical outcome of patients with inherited hemophilia who underwent antiplatelet therapy for percutaneous coronary intervention in the last 10 years.

Methods:

Retrospective chart review from a single hospital for 10 calendar years of patients with inherited hemophilia A or B and a diagnosis of coronary artery disease who received oral antiplatelet agents.

Results:

See data Table 1 below.

Conclusions:

The usage of antiplatelet therapy for coronary artery disease as well as dual-antiplatelet therapy post percutaneous coronary intervention in patients with Hemophilia A or B carries a realistic risk of bleeding. However, outside of one patient with traumatic laceration of the liver, we did not identify any life threatening bleeding events in our patients who were placed on antiplatelet therapy. Since all subjects had no more than mild to moderate factor deficiency, it would be important to see how patients with severe deficiency would do on such therapy. Further studies are needed in the future to provide evidence and clarify the overall risk versus benefit of antiplatelet therapy in hemophilia patients with coronary artery disease.

Disclosures

No relevant conflicts of interest to declare.
Most CD5+ B Lymphocytes Secrete IL-10 in Autoimmune Hemolytic Anemia/Evans Syndrome Patients
Authors: Xing Limin , Zhao Manjun , Zhu Hongli , Shao Zonghong
Published: October 2019

Objective:

Autoimmune hemolytic anemia (AIHA)/Evans syndrome is a form of immune-related cytopenia characterized by autoantibodies directed against red blood cells. A subset of B lymphocytes expressing CD5 is found in patients with AIHA/Evans syndrome. Previous studies have shown that the number of CD5+ B lymphocytes is correlated with the severity of AIHA/Evans syndrome. IL-10 can be induced in various types of human B cells, including naive, memory, and CD5+B cells, following B-cell activation by Th-cell-dependent and TLR-dependent signals. This study is to investigate the secretion function of CD5+ B lymphocytes in AIHA/Evans syndrome (ES) patients.

Methods:

25 untreated AIHA/ES patients, 28 remission AIHA/ES patients and 25 healthy controls (HCs) were enrolled in this study. The quantity of CD5+B lymphocytes which produce interleukin-10 (IL-10) (CD5+IL-10+) were detected by flow cytometry. CD5+ B lymphocytes were sorted from Peripheral blood (PB) by CytoFLEX Flow Cytometer. The expression of IL-10 mRNA in CD5+ B cells were measured by real-time PCR (RT-PCR). The concentrations of IL-10 in serum were detected by Enzyme-linked Immunosorbent assay (ELISA).

Results:

The percentage of CD5+IL-10+B cells in CD5+ B lymphocytes in newly diagnosed patients was 82.18±14.78%, which being significantly higher than that of remission AIHA/ES patients (56.68±24.39%) and HC (51.90±22.95%)(p<0.05). The percentage of CD5+IL-10+ B cells in CD5+ B lymphocytes in newly diagnosed patients was negatively correlated with their serum level of hemoglobin, C3 (P<0.05) and positively correlated with their serum level of LDH, TBIL and IBIL (P<0.05).

The expression level of CD5+ B lymphocytes IL-10 mRNA in newly diagnosed patients (49.34±22.84) was higher than that of remission patients (3.97±3.83) and HC (1.78±1.66) (P<0.05).

The concentration of IL-10 in serum in newly diagnosed patients (4.01±1.54 pg/ml) was lower than that of remission patients (5.08±1.72pg/ml) and HC (5.70±1.60pg/ml) (P<0.05).

Conclusions:

Most CD5+ B lymphocytes secrete IL-10 in AIHA/Evans patients and are positively correlated with the severity of the disease. The increased quantity of CD5+IL-10+ B cells and the decreased cytokines-secreted level maybe involved in the occurrence of AIHA/Evans syndrome.

Disclosures

No relevant conflicts of interest to declare.
The Pan-PIM Kinase Inhibitor LGB321 Strongly Increases Ibrutinib Effects in CLL, By Blocking Anti-Apoptotic Pathways and Microenvironmental Interactions
Authors: Decker Sarah , Saleem Shifa , Kissel Sandra , Zwick Anabel , Aumann Konrad , Zirlik Katja , Duyster Justus , Dierks Christine
Published: October 2019

Despite remarkable anti-tumor activity of ibrutinib in CLL, some patients develop resistance due to acquired mutations. By adding drugs which target survival pathways and the CLL protective microenvironmental interactions, we aim to increase ibrutinib efficacy and to prevent ibrutinib resistance. The three PIM kinases are involved in important disease mechanisms in CLL, with PIM1 regulating CXCR4 surface expression impacting the interaction with the protective microenvironment, and PIM2/3 affecting the apoptotic machinery by regulating BAD.

Here, we investigated the effect of the Pan-PIM kinase inhibitor LGB321 in CLL in vitro and in vivo and its cooperative effect with ibrutinib. LGB321 was highly effective in inducing apoptosis in primary CLL cells, independent of prognostic factors or microenvironmental interactions, and reduced pBAD and total BAD protein levels in CLL. In vitro combination of ibrutinib and LGB321 demonstrated strong additive effects with doubled apoptosis rates compared to single treatment. Further, LGB321 treatment blocked the CXCR4/CXCL12 axis by dephosphorylating the CXCR4 receptor on Ser339, by reducing total CXCR4 protein levels, and by blocking the externalization of the CXCR4 receptor. Concordantly, LGB321 blocked CXCR4 functions regarding migration towards a CXCL12 gradient and homing of LGB321-pretreated primary CLL cells towards the bone marrow of NOG mice.

In vivo experiments confirmed the efficacy of LGB321 in 5 different CLL xenograft studies. Transplantation of primary CLL cells into NOG mice and treatment with LGB321 for 2 weeks strongly reduced WBC counts, spleen size and spleen infiltration with human CLL cells in all 5 CLL cases, and blocked BAD as well as CXCR4 phosphorylation also in vivo.

Xenograft studies combining ibrutinib with LGB321 demonstrated much faster reduction of WBC counts than ibrutinib single treatment and strongly reduced disease burden in the double treated mice, confirming the cooperative effects of those two inhibitors also in vivo.

Our results demonstrate that LGB321 might be an effective treatment option for CLL patients by impairing PIM2/3 mediated CLL-cell survival, and by blocking the PIM1/CXCR4-mediated interaction of CLL cells with the protective microenvironment. Furthermore, LGB321 enhances ibrutinib treatment effects and might help to increase its efficacy and to avoid the development of ibrutinib resistance.

Disclosures

No relevant conflicts of interest to declare.
Quality of Life Among Adolescents with Beta-Thalassemia Major (TM) in Qatar
Authors: Nashwan Abdulqadir Jeprel , Yassin Mohamed A , Babu Ganga , Nair Sindhumole LK , Libo-on Izette , Hijazi Hothaifah , De Sanctis Vincenzo , Soliman Ashraf Tawfiq
Published: October 2019

BACKGROUND: Thalassemia is a heterogeneous group of inherited disorders of hemoglobin synthesis. It is a common disease in Mediterranean, Southeast Asia, Indian subcontinent, and Middle East countries, including Qatar.

PURPOSE: The aim of this study was to assess the quality of life (QOL) among patients aged 14 to 18 years with thalassemia major (TM) in Qatar and correlates their QOL with bio-demographic data of the patients compared to healthy controls.

MATERIALS AND METHODS: This cross-sectional study measured the QOL in adolescents with thalassemia major who were attending ambulatory units in a tertiary hospital in Qatar. Forty children and adolescents with TM and 40 healthy participants were enrolled in the study. Forty-two (52.5%) participants were males and 38 (47.5%) females. Data were collected utilizing PedsQLTM 4.0 generic core scale and were analyzed using the appropriate statistical method.

RESULTS: Children with TM had significantly lower and more variable overall quality of life score (69.1 ± 16.8) compared to healthy matched children (77 ± 12.8), (p <0.001). Both groups were not different from the physical, emotional, and social domains. Thalassemic adolescents had also a significantly lower school performance.

CONCLUSIONS: TM adversely affects the QOL of children and adolescents and this necessitates applying more efforts to help them improve and achieve a desirable quality of life. Patients with TM need more attention in schools that can be accomplished by implementing a special program for their management that needs a mutual collaboration between Ministry of Public Health (MoPH) and Ministry of Education (MoE) in Qatar.

Disclosures

No relevant conflicts of interest to declare.
Low and Very Low Bone Mineral Density Among Adult Survivors of Childhood Cancer Can be Predicted
Authors: Van Atteveld Jenneke E. , Pluijm Saskia M.F. , Ness Kirsten K. , Hudson Melissa M. , Chemaitilly Wassim , Kaste Sue C. , Robison Leslie L. , Neggers Sebastian J.C.M.M. , Van den Heuvel-Eibrink Marry M , Wilson Carmen L.
Published: October 2019

Background: Although low bone mineral density (BMD) is prevalent among adult survivors of childhood cancer, evidence-based guidance for surveillance of low BMD among survivors by dual X-ray absorptiometry (DXA) is limited. Therefore, we aimed to develop and validate prediction models for low and very low BMD based on patient and treatment characteristics that identify adult survivors who require screening by DXA.

Methods: Caucasian survivors (N=2412, median attained age, 30.9 [range: 18.1-65.9] years, 60% survivors of hematologic malignancies) enrolled in the St. Jude Lifetime Cohort (SJLIFE, development) and survivors previously treated at the Erasmus Medical Center (validation), the Netherlands (N=410, median age, 24.5 [range: 18.0-49.3] years, 73% survivors of hematologic malignancies) were evaluated with DXA to determine lumbar spine (BMDLS) and total body (BMDTB) BMD, with low and very low BMD defined as BMDLS and/or BMDTB Z-score ≤-1 or ≤-2, respectively. Backward multivariable logistic regression was used to build prediction models; performance was assessed using receiver operating characteristic curves. Diagnostic values were calculated at different probabilities. Diagnostic values were calculated at different probabilities. The formula PlowBMD=e^(β0+β1*x1+...+βn*xn)/(1+e^(β0+β1*x1+...+βn*xn)) was used to predict individual probability of low BMD (β0=intercept; βn=regression coefficient (β) of each predictor xn).

Results: Low BMD was prevalent among 52% and 44% of SJLIFE and Dutch participants, and very low BMD among 20% and 10%, respectively. The model for low BMD included male sex (β=0.50), height (β=-0.05), weight (β=-0.03), age at cancer diagnosis (β=-0.02), current smoking (β=0.19) and cranial irradiation (β=0.31) as significant predictors. Areas under the curves (AUC's) were 0.72 (95%-CI 0.70-0.74) in SJLIFE and 0.70 (95%-CI 0.65-0.75) in the Dutch cohort. Sensitivity (69.6%), specificity (63.9%) and accuracy (66.9%) were optimized at a probability of 0.50. The model for very low BMD included male sex (β=0.55), height (β=-0.05), weight (β=-0.04), cranial (β=0.31) and abdominal (β=0.22) irradiation, yielding AUC's of 0.75 (SJLIFE cohort) and 0.72 (Dutch cohort).

Conclusion: Validated models for low BMD including sex, age at cancer diagnosis, height, weight, smoking status, cranial irradiation and abdominal irradiation correctly identified BMD status in most Caucasian survivors through age 50 years. Clinical applicability can be warranted using our online calculator.

Disclosures

No relevant conflicts of interest to declare.
Prognostic Value of IPSS-R Score, Cytogenetic Aberrations, and Cytopenias on Outcome of Allogeneic Hematopoietic Stem Cell Transplant in Myelodysplastic Syndrome in GATA2 Deficiency
Authors: Scheiermann Julia , West Robert , Embree Lisa J. , Calvo Katherine R. , Shah Nirali N. , Tuschong Laura , Parta Mark , Hickstein Dennis D.
Published: October 2019

Sporadic or familial germline mutations in GATA2 frequently result in a bone marrow failure syndrome characterized by recurrent life-threatening infections, cytopenias, and progression to myelodysplastic syndrome (MDS), acute myelogenous leukemia (AML), or chronic myelomonocytic leukemia (CMML). Hematopoietic stem cell transplantation (HSCT) represents the only definitive therapy for GATA2 deficiency. However, the highly variable disease penetrance in different families, and in mutation positive individuals within a single family, make the timing of HSCT challenging. We assessed the utility of scoring patient characteristics according to the Revised International Prognostic Scoring System (IPSS-R) to predict survival after HSCT in a cohort of 51 patients with GATA2 deficiency who had progressed to MDS. Higher IPSS-R scores significantly correlated with reduced overall survival after transplant. In evaluating the components of the IPSS-R score for patients with GATA2 deficiency, cytopenias, including anemia, neutropenia, and thrombocytopenia, were important variables in placing these patients in higher risk IPSS-R categories. Since GATA2 deficiency patients have very low peripheral blood monocyte, B cell, and NK cell counts, we evaluated these cytopenias in conjunction with IPSS-R scoring. These cytopenias also had prognostic value for outcome after HSCT in patients with GATA2 deficiency. Cytogenetics, another component of the IPSS-R score, also placed patients with GATA2 deficiency into higher risk categories. In particular, patients harboring monosomy 7 had poorer survival outcomes than patients with normal cytogenetics or trisomy 8. Of note, all five patients with GATA2 deficiency and trisomy 1q successfully underwent HSCT. Clonal cytogenetic progression with the development of additional cytogenetic aberrations was a harbinger of myeloid progression and led to HSCT. In a cohort of four patients who underwent HSCT following progression to AML/CMML, there was only one survivor. Thus, HSCT is recommended in patients with GATA2 deficiency and intermediate or high-risk IPSS-R scores before the development of AML or CMML.

Disclosures

No relevant conflicts of interest to declare.
Thrombotic Thrombocytopenic Purpura in Oman; Manifestations and Outcome. Retrospective Study
Authors: Al Hashmi Khalid Nasser , Al Dowaiki Samata
Published: October 2019

Abstract:

Background: Thrombotic Thrombocytopenic Purpura( TTP) is a rare life threatening disease. It is caused by congenital deficiency of metalloprotease ADAMTS -13 or acquired autoantibodies to this enzyme which normally cleaves vWF macromolecules. This results in Microangiopathic hemolytic anemia (haemolytic anemia, low platelets, and schiztocytes), systemic microvascular thrombi formation and subsequent end organ ischemia (renal and neurological manifestations). Nowadays the early recognition and treatment modalities resulted in survival rate reaching up to >=90%. So far, in Oman there have been no studies evaluating this rare disease.

Aim: This is a retrospective study conducted in a tertiary center in Oman aims to describe the clinical and the demographic characteristics of Omani patients diagnosed of TTP, their remission rate, and survival.

Methods:. We included all patients who were diagnosed between 2006 and 2018 who meet the diagnostic criteria of TTP and aged ≥ 12 years old. Data on patients' age, gender, region, ADAMTS-13 level, clinical presentation, lab results, treatment modalities and condition on discharge are described.

Results: Total of 38 patients met diagnostic criteria of TTP during the study period. 25 Females (66%) and 13 Males (34%),with Female: Male ratio was 1.9:1. Age ranges between12-83 years (mean: 42 years). Mean age at onset: 36 years (range 1- 79 years). Majority of patients were from Governates Al-Batinah 39% and Muscat 21%. Clinical features in descending order of frequency: fever (63%), petechiae/ ecchymosis (31%), seizures (21%), high BP reading(21%), headache(15%) , confusion(15%), postpartum presentation(15%) and stroke (7.9%). The mean values at presentation are consistent with microangiopathic hemolytic anemia. Deranged RFT was found in 19 patients (50%), ADAMTS 13% was tested in 18 cases, congenital deficiency was found in 4 patients, deficiency due to autoantibodies was found in 7 patients. Treatment modality received were steroids and Plasma exchange (number of sessions ranges between 3-30 sessions, average 9 sessions). 19 cases required rituximab and 10 patients required cyclosporin. 5 patients (13%) had renal derangement requiring dialysis as inpatients, only 1 patient continued to dialyze after discharge. None of the 3 patients with documented stroke had neurological deficits on discharge. Patients who had seizures (3 patients) were free of seizures and on maintenance antiepileptic medications. Relapses after first presentation was noted in 7 patients (17%) with mean relapse ranges 1 to 7 years (mean of 3 years) after first presentation. There was 1 death with TTP in a relapsed Non- Hodgkin's lymphoma pateint. Overall survival rate was 97% at 10 years study period.

Conclusion:This is the first study that demonstrates the demographic, clinical and survival rate of TTP patients in Oman, and provides a general picture of the TTP patients in our country

Disclosures

No relevant conflicts of interest to declare.
MM-derived exosomes can promote apoptosis and inhibit proliferation of HD-CD4+T, inhibit apoptosis and promote proliferation but inhibit perforin secretion of HD-CD8+T, inhibit apoptosis and promote proliferation HD-Treg, and promotes IL-10 secretion
Authors: Fu Rong , Shao Qing , Zhaoyun Liu , Liu Hui , Chen Jin , Shao Zonghong
Published: October 2019

OBJECTIVE: To study the effects of multiple myeloma (MM)-derived exosomes on the number and function of CD4+T, CD8+T and regulatory T cells in healthy donors (HD) and MM patients.

METHODS: MM cell-derived (OPM2, U266) exosomes were extracted by ultracentrifugation and kit method and co-cultured with CD4+T, CD8+T and CD4+CD25+CD127dimT cells (Treg) from HD and MM patients sorted by magnetic beads. Flow cytometry was used to detect the apoptosis rate of T cells, level of perforin and granzyme B in CD8+T. Cell viability was detected by CCK8 kit, and IL-10 and TGF-β levels were detected by ELISA.

RESULTS: HD-CD4+T apoptotic rate was higher in the OPM2 group (6.24±1.24%) and U266 group (5.42±1.07%) than in the control group (4.37±0.96%). The HD-CD4+T viability in U266 group (85.45±6.09%) decreased compared with the control group. The apoptotic rate of HD-CD8+T was decreased in the OPM2 group (9.97±1.28%) and the U266 group (11.00±1.75%) compared with the control group (16.12±2.95%). HD-CD8+T cell viability was increased in the OPM2 group (112.63±3.88%) and the U266 group (111.70±3.62%) compared with the control group. Perforin of HD-CD8+T in U266 group (9.34±2.36%) was lower than the control group (12.76±3.31%); Perforin of MM-CD8+T in OPM2 group (6.48±1.06%) and U266 group (5.63±1.15%) decreased compared with the control group (10.55 ± 2.50%). The apoptosis rate of HD-Treg was lower in OPM2 group (15.33±3.87%) and U266 group (11.71±2.71%) than in control group (19.61±3.50%). The apoptosis rate of MM-Treg was higher in U266 group (37.29±6.54%) than the control group (29.95 ± 6.68%). The viability of HD-Treg in U266 group (173.48±34.99%) was higher than the control group. The viability of MM-Treg in OPM2 group (82.29±12.16%) was lower than the control group. The supernatant IL-10 from HD-Treg in OPM2 group (18.53±8.08 pg/ml) was higher than the control group (8.49±4.02 pg/ml). The supernatant TGF-βfrom MM-Treg was decreased in the OPM2 group (290.29±23.21pg/ml) and the U266 group (230.03±15.96pg/ml) compared with the control group (387.49±21.60pg/ml) (P<0.05).

Conclusion: MM-derived exosomes can promote apoptosis and inhibit proliferation of HD-CD4+T, inhibit apoptosis and promote proliferation but inhibit perforin secretion of HD-CD8+T, inhibit apoptosis and promote proliferation HD-Treg, and promotes IL-10 secretion.

Disclosures

No relevant conflicts of interest to declare.
Patient-Reported Outcome Labeling for Malignant Hematology Drugs Approved in the United States: 2011-2017
Authors: Zettler Marjorie E , Basch Ethan , Nabhan Chadi
Published: October 2019

Introduction: Patient-reported outcomes (PROs), defined as any report on the status of a patient's health condition that comes directly from the patient without interpretation by anyone else, play an increasingly important role in drug development. In 2009 the FDA issued final guidance on using PRO measures to support labeling claims, to incorporate the patient perspective into the drug approval process. The 21st Century Cures Act emphasizes PROs as a differentiating element in the FDA approval process of new drugs, beyond traditional clinical outcome measures. Further, recent data has shown that intervention based on PROs can improve survival in metastatic cancers (Basch et al; 2017). The incorporation of PROs into the labeling of new drugs for malignant hematology disorders has not been studied and is the subject of this investigation.

Methods: We reviewed the FDA's Novel New Drug Summaries (2011-2017) to identify drugs approved for malignant hematology indications. Drug approval packages and product labeling were accessed via the Drugs@FDA database and analyzed for PRO endpoints, measures, and labeling claims. Clinical trial designs and published study results were retrieved via the ClinicalTrials.gov website and PubMed.

Results: Of 250 novel drugs approved by the FDA between 2011 and 2017, 22 (8.8%) were approved for malignant hematology indications. Interestingly, only 1 had PRO-based claims in their labeling, even though 13 of the 22 drugs (59%) collected PRO data in pivotal trials that led to their approval. Notably, the proportion of malignant hematology trials assessing PROs has increased over time, with 4 of the 5 drugs approved in 2017 for malignant hematology indications evaluating PROs in their development programs, compared with 9 of the 17 drugs approved in the preceding 6 years (80% vs. 53%). PROs evaluated included generic instruments such as the EQ-5D, and disease-specific instruments such as the EORTC QLQ-C30 (see table). Reasons cited for rejection of PRO data inclusion in drug labeling were single arm trial design, excessive missing data, statistical issues, and use of an inappropriate PRO instrument.

Conclusions: While the FDA encourages PRO data submission as part of the new drug approval process, and although more than half of all malignant hematology drugs approved in the past 7 years assessed PROs during development, only 1 was able to successfully acquire labeling claims. Whether this is due to lack of PRO expertise on clinical development teams or absence of strong regulatory guidance on how best to implement PROs remains unknown and requires further research. Designing strategies to develop, validate and report PROs effectively is needed to meet regulatory requirements and enhance patients' voices in their own care.

Table. Table.

Disclosures

Nabhan: Cardinal Health: Employment, Equity Ownership.