bluuebunny/crossref_metadata_2025 huggingface dataset:
R-COMP Regimen in B Cell Non Hodgkin Lymphoma Patients with Cardiovascular Comorbidities
Authors: Annibali Ombretta , Chiodi Francesca , Sarlo Chiara , Cenfra Natalia , Tomassini Simona , Mecarocci Sergio , Cimino Giuseppe , Avvisati Giuseppe
Published: October 2019

Backgroung: Anthracyclines-based regimens remain the gold standard for the treatment of Lymphomas, although the associated cardiac toxicity may limit their use, especially in frail and elderly patients.

Patients and Methods: From October 2008 to January 2015 we treated 51 newly diagnosed patients B cell with poor-risk non Hodgkin lymphoma and cardiovascular comorbidities using the R-COMP regimen (rituximab, cyclophosphamide, non-pegylated liposome-encapsulated doxorubicin, vincristine and prednisone). Median age was 72 years (range:46-82 yrs ; 62% ≥ 70 years). As for histology, 26/51 (50%) were Diffuse Large B-cell Lymphoma; 10/51 (20%) Follicular Lymphoma; 8/51 (16%)mantle cell lymphoma; 5/51 (10%) Nodal Marginal Zone Lymphoma; 2/51 (4%) other B-cell indolent Lymphoma. IPI and FLIPI prognostic scores were Intermediate to High in the majority (39/51 = 76%) of the patients. Stage III and IV according to Ann-Arbor staging system was present in 40/51 patients (78%). The median age adjusted Charlsons comorbidity index was 6 (range: 3 to 11). Cardiovascular risk factors were considered: hypertension (39/51pts = 76%), a history or recent acute myocardial infarction (9/51 pts = 17%) and Atrial fibrillation (4/51 = 8%). According to National Institute of Aging/National Cancer Institute (NIA/NCI index), a large portion of patients (39%) presented high-impact conditions mainly consisting of ischemic and arrhythmic diseases, under active treatment. Treatment was well tolerated and toxicities were limited grade III/IV cytopenia.

RESULTS: Complete remission was achieved in 37/51 (72%) and partial response in 8/51 (15%). The remaining 12% of patients had a progressive disease . As of July 2015, after a median follow up period of 25 months (range 3-79), the OS, RD, EFS, TTF were not reached ( Figure 1, panel A,B,C respectively). In particular, at 5 years from the treatment starting, OS and RD are both 70% and EFS is 54% .Cardiac toxicity was observed in one patients who died for pulmonary edema, while two patients developed arrhythmias.

Conclusions: This study confirm the efficacy and tolerability of R-COMP regimen in elderly patients with cardiovascular comorbidities.

Figure 1. Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Up-Regulation of CAR-T Cells after G-CSF and Dexamethasone Treatment in a Defuse Large B Cell Lymphoma Patient Receiving 4SCAR19 T Cell Therapy
Authors: Jing Hongmei , Chang Lung-Ji , Chen Mingyi , Bao Fang , Wang Jing , Wang Wenming , Liu Yuanyuan , Kuo Hao-Hsian , Liu Yuchen , Dong Lujia
Published: October 2019

A 37-year-old woman was diagnosed with DLBCL, GCB origin, positive with CD20, Bcl-2, Bcl-6, CD10 and negative with CD3, CD5, ,D21, CD23, CyclinD1. PET-CT scan showed that her lymph nodes were broadly involved. Based on PET-CT, the stage was IIIA and aaIPI score was 1. After eight cycles of R-CHOP (rituximab, cyclophosphamide, epirubicin, vincristine, and prednisone), the tumor regressed to a small retroperitoneum lesion with SUV 2.0, and she received another four cycles of rituximab for maintenance. The tumor relapsed one year later with resistance to ESHAP after she had failed multiple alternative treatments including GA101 trial, 3x DICE, MTX1g, 2x GEMOX, and 2x EPOCH. She was enrolled in a CD19-CAR (chimeric antigen receptor) T cell pilot study in January 2015. Her T cells were apheresis collected and transduced with a 4th generation, apoptosis-inducible, safety-engineered lentiviral CAR: CD19- scFv/CD28/CD137/CD27/CD3ζ-iCasp9 (4SCAR19). A personalized conditioning regimen was given based on the patient's history to chemotherapy: cyclophosphamide 500mg/d d1-3, and fludarabine 50mg/d d1-4, 40mg d5. Two days later, she received infusions of a total dose of 1.27×108 of the 4SCAR19 T cells (2x106/kg). At day 7 (D7) after infusion, she developed a fever over 39o C, which lasted for 7 days; this was controlled with NSAIDs (Non-Steroidal Anti-inflammatory Drugs). The tumor in her lymph nodes began to shrink 5 days after CAR-T infusion and she achieved nCR (near complete response) after 30 days. In a follow up PET/CT 3 months after CAR-T infusion, there was only one suspected retroperitoneum lesion (SUV 3.8) in the whole body, but it was indiscernable whether the SUV signal was tumor or T cell related. We monitored the peripheral blood CAR-T cell counts by qPCR and detected 0.03%, 0.07%, 0.3%, 9.3%, 0.01%, 1.57% on D7, D13, D48, D69, D84 and D112, respectively. In attempt to perform an autotransplantation to pursue a cure, she was mobilized twice with G-CSF on D68-D72, D110-112, plus dexamethasone 20mg/d on D101-102 and 15 mg/d on D110-111. Unexpectedly, the CAR-T cells in the patient peripheral blood increased to 9% after the first mobilization, and to 18% after the second mobilization. To this date, the patient has remained in nCR after 4SCAR19 therapy (January 23 to July 23, 2015). To our knowledge, this is the first report of evident CAR-T cell boost associated with G-CSF plus dexamethasone treatment. Further investigation is warranted to understand the molecular mechanisms behind such a favorable CAR-T therapy outcome in a terminal DLBCL disease.

Disclosures

Kuo: America Yuva Biomed: Employment. Liu:America Yuva Biomed: Employment. Dong:America Yuva Biomed: Consultancy.
Correlation Between Tumor-Associated Macrophage and Inflammatory Factors in Lymphoma Patients
Authors: Sun Xiaolin , Ni Jun , Wang Hong , Sun Xin , Wang Cuimei , Wu Wei , Gu Jian , Depei Wu
Published: October 2019

Introduction

To study the expression of tumor associated macrophage (TAMs), inflammatory cytokines IL-1β (interleukin-lβ), TNF-α (tumor necrosis factor-α), IL-6 (interleukin 6) and IL-10 (interleukin 10) and its significance in different types of lymphoma in lymph node pathological tissue.

Methods

106 cases of clinical pathology specimens of patients were collected from different types and different staging classifications. The expressions of tumor-associated macrophage and interleukin-1β (IL-1β), tumor nec.rosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10) in lymphoma tissues were detected by immunohistochemical staining;

Results

Both CD68 and CSF-1R showed high expression in lymphoma tumor tissues. The positive rates of CD68 and CSF-1R in Ⅳ period patients was higher than Ⅱperiod (P <0.05), and the level of CSF-1R in high-risk group was higher than low-risk group (P <0.05). The expression of CD68 was positively correlated with CSF-1R. The inflammatory cytokines such as IL-1β, TNF-α, IL-6, IL-10 were highly expressed in tumor tissues; the positive rate of IL-1β in lymphoma IV period was higher than periodⅡ(P <0.05) and the level of IL- 6 in high-risk group were significantly higher than the low-risk group (P <0.05).The expression of tumor-associated macrophage phenotype CD68 in lymphoma tissues was positively correlated with inflammatory cytokines IL-1β, TNF-α, and IL-6 (P <0.05).

Conclusion

1.CD68, tumor-associated macrophage phenotype, and CSF-1R were highly expressed in lymphoma tissues

2.The expression of CD68 was positively correlated with inflammatory cytokines.

3.TAMs were involved in tumor inflammatory microenvironment.

Funded by Jiangsu Provincial Special Program of Medical Science (BL2012005)

Disclosures

No relevant conflicts of interest to declare.
Survey of Qol (quality of life) on Patients Receiving Tyrosine Kinase Inhibitors for Chronic Myeloid Leukemia: Korean CML Working Party Group
Authors: Kim Kyoung Ha , Kim Dae-Young , Kim Sung-Hyun , Kim Yeo-Kyeoung , Kim Inho , Kim Hee-Jin , Sohn Sang Kyun , Lee Jeon-Ok , Cheong June-Won , Jung Chul Won , Cho Young-Uk
Published: October 2019

Background

With imatinib therapy, the survival of patients with CML has improved dramatically. In recent years, second generation TKI (2G TKI) led to more increased responses. Despite the demonstrated superiority of efficacy and toxicities as previous treatments, several adverse events of TKIs remain. Because CML treatment is lifelong, it is important for patients to maintain adherent to treatment. Correlations between poor adherence to TKI treatment and adverse events and quality of life (QoL) have been demonstrated among CML patients in clinical settings. The FACT-leu is a validated tool that measures leukemia-specific and more general QoL concerns. However, there are few studies about QoL of CML patients in Korea. Therefore, we evaluated the QoL of CML patients treated with TKIs.

Methods

The CML working party of the Korean society of Hematology was produced by the questionnaire included the questions of FACT-leu. We distributed the questionnaire over the online or offline. QoL of patients with CML was analyzed using these questionnaires.

Results

A total of 384 patients were surveyed between March 2014 and June 2014. Male was 234 (60.94%) patients. Median age was 47 (range 12-71) years. Current taking medication at survey were imatinib (n=197), dasatinib (n=71), nilitinib (n=88), radotinib (n=28). Patients with high compliance of TKI (taking 90% or more drugs) were 88.28% and lower compliance (taking 70% or less drugs) were 1.82%. In queries about poor adherence to TKI therapy, common causes were forgetfulness (18.49%) and adverse events (8.59%). FACT-Leu scale was higher in patients with 2G TKIs than imatinib (122.9 ± 22.65 vs 116.0 ± 25.00 (p=0.005)). Female, older patients, patients with CML in longer time showed meaningful worse QoL in FACT-leu scale. There was no significant difference between dasatinib (122.2±25.5) and nilotinib (124.6±22.06) in FACT-leu scale.

Conclusion

In 2G TKI era, QoL of CML patients seemed that is better than before. Additional prospective studies are warranted using adequate measuring tool of QoL.

Disclosures

No relevant conflicts of interest to declare.
Labile Plasma Iron (LPI), More Practical and More Sensitive to Iron Overload in Myelodysplastic Syndromes
Authors: Chang Chunkang , Gu Shucheng , Guo Juan , Zhao Youshan , Fei Chengming , Li Xiao
Published: October 2019

Background: In order to gain an insight into LPI in iron metabolism microenvironment in MDS.

Materials and Methods: We performed ELISA, quantitative RT-PCR, flow cytometry, MRI T2* assays to test LPI, iron biochemical parameters and LIC among 22 MDS patients.

Results: LPI has a statistical difference (P<0.001 by ANOVA), which decreased gradually, among three groups, while no difference was found in ASF (P=0.086 by ANOVA). After DFO treatment, serum hepcidin expression increased from 301.26±59.78¦Ìg/L to 340.33±49.78¦Ìg/L (P=0.032), while hepcidin/ASF was up-regulated gradually from 0.16±0.08 to 0.22±0.03 (P=0.045). APAF-1 expression (P=0.047) and erythroid apoptosis rate (P=0.009) decreased significantly, respectively. No statistical difference was found in EPO (P=0.247) and GDF15 expression (P=0.172). LIC dropped from 9.83±4.84mg/g dry weight to 6.28±4.01mg/g dry weight (P<0.001). No significant difference was found in cardiac T2* (P=0.594). LPI has a closer connection to LIC than ASF (r=0.739, P<0.001 vs. r=0.321, P=0.034). (Figure1)

Conclusion: LPI seems to be a real-time indicator which reflects body iron loading status instantaneously. Despite the limited knowledge available on LPI speciation in different types and degrees of IO, LPI measurements can be and are in fact used for identifying systemic IO and for initiating/adjusting chelation regimens.

Before DFO treatment, a patient had an abnormal liver function. Along with iron chelation therapy, his LPI decreased rapidly. Meanwhile, his ALT dropped and his hemoglobin increased gradually. Furthmore, his LIC decreased after receiving DFO treatment.

Lane a: LIC was 9.97mg/g dry weight on week 0.

Lane b: LIC was 5.81mg/g dry weight on week 26.

Figure 1. The improvement of ALT, hemoglobin and LIC Figure 1. The improvement of ALT, hemoglobin and LIC Figure 2. Figure 2.

Disclosures

No relevant conflicts of interest to declare.
Invasive Fungal Infections in Patients with High-Risk MDS and AML Treated with Azacitidine
Authors: Pomares Helena , Arnan Montserrat , Sánchez-Ortega Isabel , Sureda Anna , Duarte Rafael F.
Published: October 2019

Introduction: The incidence of invasive fungal infections (IFI) and need for antifungal prophylaxis in patients with high-risk myelodysplastic syndromes and acute myeloid leukemia (MDS/AML) treated with hypomethylating agents is currently unknown.

Methods: We retrospectively analyzed the incidence of IFI in all MDS/AML patients receiving azacitidine (75 mg/m2/day 7 days every 28-day cycle) in our center between June 2007 and June 2015. IFI diagnosis follows the European Organization for Research and Treatment of Cancer and Mycosis Study Group (EORTC/MSG) 2008 Criteria. Patients did not receive antifungal prophylaxis.

Results: One hundred and twenty one consecutive unselected patients (86 MDS and 35 AML; Table 1) received a total of 948 cycles of azacitidine in this series (median 5, range 1-43). One hundred and ten (91%) received azacitidine frontline and 11 (9%) following refractoriness or relapse after previous intensive chemotherapy. Seventy one patients received ≥4 azacitidine cycles. Forty two (35 %) patients had complete medullar and cytogenetic responses. Two hundred and seventy four azacitidine cycles (28%) in 49 patients (40%) were administered during and/or led to prolonged grade IV neutropenia (<0.5x109/L ³10 days), and caused febrile neutropenia episodes in 121 cycles (12% of total, 44% of neutropenic cycles) in 45 patients (37%), range 1-6 per patient. Four IFI occurred in these patients: one possible, two probable and one proven (Table 2). The incidence of IFI was 0.42% per treatment cycle and 3.3% per patient treated for the overall series, and 0.72% per treatment cycle and 4.1% per patient treated among those with prolonged severe neutropenia. Of note, all cases of IFI occurred in either the first or the second azacitidine courses. Two patients died from IFI, leading to an IFI-related mortality rate of 1.65% per patient treated and 0.21% per treatment cycle. The numbers needed to treat with primary prophylaxis, even for an ideal agent that could potentially prevent all IFI, are well in excess of 100 courses of azacitidine and well over 20 patients throughout their treatment course, to prevent one single IFI in these patients, even among those with neutropenia.

Conclusion: Our data show that patients with MDS/AML treated with azacitidine have a very low risk of IFI, including those with concurrent severe prolonged neutropenia. Primary antifungal prophylaxis should not be recommended for this subset of patients. This very low risk of IFI is a potential additional benefit from treatment of MDS/AML with hypomethylating agents, compared to conventional intensive chemotherapy. Prospective studies are needed to better address this issue.

Disclosures

Sureda: Takeda: Consultancy, Speakers Bureau.
The Retinoid Derivative Drug, Acitretin, Modulates CD38 Expression on MEC-1 Cells and Primary Chronic Lymphocytic Leukemia (CLL) Cells and Reduces Migration in Response to the Chemokine CXCL12
Authors: Elsir Mohammed Sally , Murphy Philip , Quinn John , Thornton Patrick , Bergin Stephen , Hennessy Bryan
Published: October 2019

Background: CD38 is a cell surface receptor and ectoenzyme expressed in a variety of haematological malignancies. In CLL, CD38 expression is a marker of poor prognostic disease and is now attracting growing interest as a target molecule for anti CLL therapy.

Retinoids modulate CD38 expression due to the presence of the Retinoic Acid Response Element (RARE) DNA sequence in the promotor region of the CD38 gene. In this study we demonstrate that the retinoic acid derivative, acitretin, upregulates the expression of CD38 on the MEC-1 cell line and on primary CLL cells from CD38 positive patients. We propose that, by doing so, Acitretin may render these cells more susceptible to the actions of anti CD38 monoclonal antibody drugs and thus have a potential role as an adjunct to such therapies.

Methods: Primary CLL cells were freshly isolated from whole blood of CLL patients using the FICOLL density centrifugation technique. The CLL like cell line MEC-1 was obtained from DSMZ, Germany.

MEC-1 cells and cells from three CD38 negative and five CD38 positive patients were treated with 10micM acitretin for 24, 48 and 72hrs. CD38 expression was measured by FACS analysis at baseline and at the different time points. Results are expressed as fold increase in Mean Fluorescence Intensity (MFI).

Migration assays were performed using 2x10^6 cells per well incubated with 10microM acitretin overnight in 1%FBS. Cells were subsequently transferred to chamber inserts overlying 10%FBS media containing 200ng/ml CXCL12 in all wells, except negative controls. Cells were then allowed to migrate for 4hrs. Migration was calculated as percentage of cells in lower chamber to upper chamber.

Results: Acitretin increased the expression of CD38 on MEC-1 cells by a mean of 7.4 fold and on cells from CD38 positive patients by a mean of 3.4 folds (n=5). Acitretin, however, did not modulate CD38 expression levels on primary CLL cells from CD38 negative patients (n=3).

Migration of primary CLL cells from 11/14 patients toward the CXCL12 chemokine was reduced in response to Acitretin treatment as compared to untreated controls. Mean migration of treated cells was 12.3% versus 27.3% mean migration in untreated controls.

Conclusion: Our findings re-affirm the potent effect of retinoid drugs as upregulators of CD38 expression and suggest that their use in CLL therapeutics in conjunction with anti CD38 monoclonal antibodies should be explored. We also propose that acitretin appears to exert a favourable blocking effect on CLL cell migration towards micro-environmental chemokines such as CXCL12, warranting further evaluation.

Disclosures

Murphy: Celgene: Honoraria.
Mutations in the Second Linker of KLF1 Cause Congenital Non-Spherocytic Hemolytic Anemia Due to Global Reduction of In Vivo DNA-Binding Affinity
Authors: Huang Stephen , Gillinder Kevin R. , Sorolla Annabel , Whitelaw Emma , Perkins Andrew C
Published: October 2019

The mommeD45 mutation generates an amino acid transversion (H350R) within a conserved linker peptide between zinc fingers two and three of Klf1 (linker 2). Klf1H350R/H350R mice have mild compensated microcytic anaemia 1. Mice Carrying the H350R mutation were interbred with Klf1+/- mice. Klf1H350R/-mice have severe perinatal haemolytic anaemia, jaundice and marked splenomegaly. Haematological evaluation of these mice shows similar phenotypes to human patients who are compound heterozygotes for null and linker 2 mutations in KLF12. Analysis of Klf1H350R/- fetal liver by flow cytometry showed an increase in circulating immature CD71+ Ter119+ erythroblasts. In the bone marrow, a marked reduction in mature (Cd71- Ter119+) red blood cells was observed. Flow cytometric analysis of the spleen from Klf1H350R/- animals revealed an expansion of erythroid cells consistent with extramedullary erythropoiesis. ChIP-seq for Klf1 in 14.5DPC fetal liver from Klf1H350R/H350R mice revealed no loss in specificity when compared to wildtype Klf1, but a global reduction in affinity. Affinity measurements of recombinant zinc finger domains in vitro will be presented. By RNA-seq, we observed significantly lower expression of Klf1 target genes in mice homozygous for the H350R mutation compared to mice carrying a wildtype allele. And this correlates with reduced DNA binding observed in ChIP-seq and in vitro assays. Previous studies of the linkers in C2H2 zinc finger transcription factors have revealed their necessity as structural and regulatory components for the C2H2 class of transcription factors. Our results show the second linker of Klf1 plays an indirect role in DNA-binding and does not act just as a spacer for the zinc fingers.

References:

1. Sorolla A, Tallack MR, Oey H, et al. Identification of novel hypomorphic and null mutations in Klf1 derived from a genetic screen for modifiers of alpha-globin transgene variegation. Genomics. 2015;105(2):116-122.

2. Viprakasit V, Ekwattanakit S, Riolueang S, et al. Mutations in Kruppel-like factor 1 cause transfusion-dependent hemolytic anemia and persistence of embryonic globin gene expression. Blood. 2014;123(10):1586-1595.

Disclosures

Perkins: Novartis Oncology: Honoraria, Membership on an entity's Board of Directors or advisory committees; Bristol-Myers Squibb: Honoraria.
The TAL1 Short Isoform Generated By PRMT1-Mediated Alternative RNA Splicing Promotes Erythroid Differentiation
Authors: Jin Shuiling , Tran Ngoc-Tung , Su Hairui , Huang Suming , Zhao Xinyang , Liu Yanyan
Published: October 2019

The isoforms of key transcription factors in hematopoiesis such as TAL1, GATA1 and RUNX1 are generated through alternative RNA splicing regulated by the PRMT1-RBM15 axis (Zhang et al. 2015). The functions of short isoforms of GATA1 (GATA1s) and RUNX1 (RUNX1a) are well characterized, yet it is unknown how the short isoform of TAL1 (TAL1s) regulates hematopoiesis. In this presentation, we report that the short isoform of TAL1, i.e. TAL1s, is generated via alternative RNA splicing as detected by isoform specific real-time PCR reactions using RNA isolated from leukemia cell lines and primary human cord blood cells. RBM15, an RNA binding protein, which is involved in chromosome translocation to produce RBM15-MKL1 fusion protein in acute megakaryocytic leukemia, regulates the alternative RNA splicing of TAL1. RBM15 promotes the production of full-length TAL1 mRNA, while reduction of RBM15 protein level via PRMT1-mediated degradation pathway favors the production of TAL1s. RBM15 directly binds to intronic regions on TAL1 pre-mRNA. Binding of RBM15 is responsible for recruiting SF3B1-associated RNA splicing complex. Given that PRMT1 senses the hypoxia status of hematopoietic cells, the changing of TAL1s/TAL1fl ratio by PRMT1 activity may be an adaptive response of hematopoietic cells to hypoxia status. The short form TAL1s still contains the helix-loop-helix DNA binding domain but not the N terminal regions upstream of the DNA binding domain. Thus, the TAL1s may act as a dominant negative mutant of the full-length TAL1fl to block TAL1fl-regulated transcription. We demonstrated that overexpression of TAL1s not the full-length TAL1promotes the erythroid differentiation of K562 cells. Although TAL1 gene is required for both erythroid and megakaryocyte differentiation at early stage of hematopoiesis, TAL1s does not promote megakaryocyte differentiation. Therefore, fine-tuning the TAL1 isoforms by the PRMT1-RBM15 axis determine the cell fate of a MEP progenitor cell. Using immunoprecipitation assays and mass spectrometry analysis, we identified proteins specifically associated with the N terminal region of TAL1. How unique TAL1s-associated transcriptional regulatory complex plays in erythroid differentiation will be discussed in the presentation in comparison with the Tal1fl-asociated protein complex. In summary, our findings stratify another new layer of regulation by PRMT1, which relays extracellular signals (such as hypoxia signal) to transcriptional regulatory program. Given that PRMT1 is often constitutively highly expressed in leukemia cells, how overproduction of short form TAL1 interferes with normal hematopoiesis may help to explain the molecular mechanisms of many hematological malignancies associated with dysregulation of TAL1 expression.

Disclosures

No relevant conflicts of interest to declare.
Universal Screening for G6PD Deficiency Detects More Cases Than Selective Testing
Authors: Segal Ilan , Aviner Shraga , Abu Rmaileh Ashraf , Cohen Ornit , Sayag Avraham , Zangen Shmuel
Published: October 2019

Background: Testing for glucose-6-phosphate dehydrogenase (G6PD) deficiency is traditionally restricted to jaundiced neonates, as well as to those whose risk factors provide sufficient grounds to suspect the deficiency. We assumed that in a multi-ethnic, multicultural society, routine neonatal screening for G6PD deficiency would detect more affected neonates.

Methods: An observational, population-based, cohort study was designed to compare the incidence of G6PD deficiency between two groups of neonates using a validated qualitative enzyme test. Incidence of G6PD deficiency was calculated retrospectively for one group of neonates born between January 2005 and July 2012 (i.e., epoch 1), when only "at-risk" newborns were tested for the deficiency (i.e., group 1). Incidence was also calculated for a second, prospective group of neonates, who were all to be born between August 2012 and April 2014 (i.e., epoch 2), and were all screened for the deficiency irrespective of risk factors.

Results: A total of 39,268 live-born infants were included in our study. In epoch 1, 6.8% of all newborns were tested for the deficiency, compared to about 87% in epoch 2. The incidence of G6PD deficiency was 0.4% (119/29,332) and 1.6% (156/9,936) in epoch 1 and epoch 2, respectively (p<0.05). Among females, there was a 6.6-fold increase in incidence of G6PD deficiency between epoch 1 (0.08% [12/14,410]) and epoch 2 (0.53% [26/4,881]), while among males there was a 3.4-fold increase in incidence between epoch 1 and epoch 2 (0.72% [107/14,922] and 2.47% [125/5055], respectively).

Conclusions: As universal screening for G6PD deficiency detects more affected newborns than the selective, risk-factor-based testing does, we suggest that validation of our study and cost-effectiveness analyses may further the aim of introducing universal screening for G6PD deficiency for neonates born in multi-ethnic, multicultural society setting.

Disclosures

No relevant conflicts of interest to declare.
ANX005, an Inhibitory Antibody Against C1q, Blocks Complement Activation Triggered By Cold Agglutinins in Human Disease
Authors: Gertz Morie A. , Qiu Haiyan , Kendall Laura , Saltarelli Mario , Yednock Ted , Sankaranarayanan Sethu
Published: October 2019

Cold agglutinin disease (CAD) is an autoimmune hemolytic anemia characterized by the presence of autoantibodies (cold agglutinins) that bind to red blood cells (RBC) at low temperatures. Cold agglutinin binding to RBCs results in antibody-mediated classical complement activation with deposition of complement C4 and C3 fragments onto the RBC surface. Complement-opsonized RBCs are removed from the circulation by macrophage-driven phagocytosis in the liver or spleen and via intravascular hemolysis following assembly of the terminal lytic complex (C5b - C9), together resulting in clinical anemia.

C1q is the initiating molecule for cold agglutinin-mediated complement activation on the surface of human RBCs. We hypothesized that directly blocking C1q recruitment onto cold agglutinin-sensitized RBCs will prevent complement activation and opsonization and reduce hemolysis. To this end, we have developed a humanized monoclonal antibody (ANX005) that binds with high-affinity (~10 pM) to C1q and blocks classical complement activation & hemolysis in an in vitro sheep RBC assay. We evaluated the impact of ANX005 on hemolysis and complement deposition on human RBCs that were pre-sensitized with sera from CAD subjects. ANX005 showed a dose-dependent reduction in hemolysis using both individual and pooled CAD sera as the source of cold agglutinin. We further demonstrated that C1q blockade led to a robust reduction in C4 and C3 fragment deposition onto human RBCs. These results demonstrate that C1q inhibition is an effective way to impede C4 and C3 activation and downstream assembly of the lytic complex in sera from CAD patients, and support the clinical development of ANX005 in CAD and other antibody-mediated diseases.

Disclosures

Gertz: Prothena Therapeutics: Research Funding; Sandoz Inc: Honoraria; NCI Frederick: Honoraria; Celgene: Honoraria; Med Learning Group: Honoraria, Speakers Bureau; Research to Practice: Honoraria, Speakers Bureau; Alnylam Pharmaceuticals: Research Funding; Novartis: Research Funding; Ionis: Research Funding; Annexon Biosciences: Research Funding; GSK: Honoraria. Qiu:Annexon Biosciences: Employment, Equity Ownership. Kendall:Annexon Biosciences: Employment, Equity Ownership. Saltarelli:Mallinckrodt: Equity Ownership; Abbvie: Equity Ownership; Annexon Biosciences: Employment, Equity Ownership, Patents & Royalties. Yednock:Annexon, Inc: Employment, Equity Ownership. Sankaranarayanan:Annexon Biosciences: Employment, Equity Ownership.
The Impact of a Multidisciplinary Sickle Cell and Gynecology Clinic
Authors: Tasha Vardya A
Published: October 2019

Background: Contraception and pregnancy planning are critically important for patients with sickle cell disease (SCD). However, rates of contraceptive utilization are often suboptimal and unplanned pregnancy remains a problem among this patient population. In an effort to improve care, a multidisciplinary hematology and gynecology clinic was initiated at University Hospitals Case Medical Center in which women of reproductive age received comprehensive care in single visits.

Objective: To determine the impact on pregnancy, contraception use and sexually transmitted infections (STIs) diagnoses of a multidisciplinary hematology and gynecology clinic for women with SCD.

Methods: A retrospective cohort study of women seen in University Hospitals Hematology Clinic for sickle cell disease from 2011 to 2015. Baseline demographics as well as the subsequent outcomes of pregnancies, contraception use and STIs were collected. Total differences in outcomes before and after the multidisciplinary clinical were evaluated with Fisher's exact test. Longitudinal differences in pregnancies and STIs were analyzed with time-to-event analyses. For outcomes of interest p charts were constructed with upper and lower control limits.

Results: A total of 133 women total were identified as reproductive age women with sickle cell disease from 2011 to 2015. The percentage of SCD patients using birth control before and after the multidisciplinary clinic increased from 12.3% to 37% (p=0.0001). The p chart for contraception use is provided bellow and was highly significant. No significant differences in pregnancy or STIs were identified in time-to-event analysis and p charts, though the total number of STIs before and after the intervention was limited (eighteen).

Conclusion: Incorporating a multidisciplinary approach with combined hematology and gynecology care for women with sickle cell disease has the potential to improve access to contraception use in this population.

Figure Figure.

Disclosures

No relevant conflicts of interest to declare.
Clinical and Biological Determinants of Bleeding Manifestation in Congenital Factor XI Deficiency; A Systematic Review
Authors: Drakulic Milana , Chai-Adisaksopha Chatree , Iorio Alfonso
Published: October 2019

Background: Congenital factor XI (FXI) deficiency is a rare inherited bleeding disorder caused by mutation of the FXI gene. Clinical manifestation and bleeding severity of FXI deficiency are varied. To our best knowledge, there are no studies investigating the predictors of bleeding in these patient population. Our study aims to investigate the clinical presentation of congenital FXI deficiency and to identify clinical, biological, and genetic determinants of bleeding.

Methods: We conducted a systematic search in MEDLINE and EMBASE databases from inception to March 2016. We included the studies of congenital FXI deficiency. In terms of study design, we included prospective or retrospective studies as well as case series and case reports. We collected clinical characteristics of the patients including age, sex, family history of FXI deficiency, family history of bleeding, and site of bleeding. We also collected the laboratories including baseline aPTT, FXI level, and genetic test results.

Results: There were 141 studies included in this review, involving 728 patients with reported congenital FXI deficiency. Forty-three percent of the patients were male. Clinical characteristics of the patients was demonstrated in Table 1. Of these, 421 (51.84%) patients had bleeding tendency. Surgical bleeding was the most common bleeding manifestation (31.88%), following bymucocutaneous bleeding (13.35%) and hypermenorrhea (9.11%). Factor XI gene mutation was found in 422 patients (57.97%). Type II mutation was the most common reported mutation (45.19%). With regards tozygosity of patients with gene mutation, 43.94% of patients had heterozygosity of factor XI gene mutation. Univariate analysis revealed that patients with baseline factor XI level less than 25% were associated with higher risk of bleeding (odd ratio 1.48 (95% confidence interval [CI]; 1.05 to 2.09). In addition, patients with combined mutation were at higher risk of bleeding when compared to patients with single mutation (odd ratio 2.83 (95% CI; 1.09-7.39).

Conclusion: Our study found that patients with factor XI deficiency has mild bleeding manifestation. Half of the patients do not experience bleeding tendency. Surgical bleeding is the most common presentation in these patient population and likely to be the first bleeding symptom. Baseline factor XI level less than 25% is the predictor of bleeders as well as combined gene mutation.

Table 1 Characteristics of patients with congenital factor XI deficiency Table 1. Characteristics of patients with congenital factor XI deficiency

Disclosures

No relevant conflicts of interest to declare.
HSP27: A Therapeutic Target in Myelofibrosis
Authors: Sevin Margaux , Pernet Nicolas , Vitte Franck , Ramla Selim , Sagot Paul , Martin Laurent , Villeval Jean Luc , Lacout Catherine , Plo Isabelle , Garrido Carmen , Girodon Francois , de Thonel Aurelie
Published: October 2019

Myelofibrosis (MF) is the most aggressive myeloproliferative neoplasms (MPN) with the highest degree of morbidity and mortality, including progressive bone marrow fibrosis resulting into bone marrow failure. JAK2 kinase inhibitors have been successfully used for a few years in MPN and more particularly for MF treatment. Despite their beneficial effects on spleen size and symptoms, JAK2 inhibitors induce low molecular and survival responses underscoring the urgent need for other therapeutic approaches. Recently, heat shock protein 90 (HSP90) - known to stabilize JAK2 - has been reported as a promising therapeutic target in MPN. However HSP90 inhibitors show toxicity and induce the expression of stress-inducible proteins like HSP70 and, most likely HSP27 as previously shown in other cancers. In addition, we and others have shown that HSP27, was strongly expressed in patients with idiopathic pulmonary, lung or kidney tubulointerstitial fibrosis, underlying a relevant role of HSP27 in fibrotic processes. Taking into account both the beneficial effects of HSP inhibitors in leukemia and in MPN, and the possible implication of HSP27 in fibrosis, we have evaluated in this work, the status of HSP27 in MF patient's samples and assess the effectiveness of an HSP27 oligonucleotide inhibitor called OGX-427.

In this study, we first assessed the extracellular and intracellular level of HSPs from MF patients by ELISA, flow cytometry and by immunohistochemistry. We observed for the first time a specific increase in both intracellular and extracellular HSP27 in CD34+ circulating hematopoietic progenitor cells (n=9-16; P=0.0097) and in the sera of patients (n=24-27; P<0.0001) with MF compared with healthy donors, respectively. Moreover, we identified the presence of HSP27 in the bone marrow's MF patients.

We then investigated the in vivo impact of OGX-427, a specific inhibitor of HSP27, or an oligonucleotide control in a murine TPO-induced MF mouse model. The use of OGX-427 limited the progression of the disease in our MF mouse model (n=9). In particular, OGX-427 was associated with a marked reduction in both the spleen weight and size. Also, we noted a decrease of megakaryocyte hyperplasia in the bone marrow accompanied by a visible restoration of spleen structure and lymphoid white pulp territories with OGX-427.

Taking altogether, our results support a key role of HSP27 in the pathophysiology in MF and highlight the potential therapeutic benefit of HSP27 inhibitors in this disorder.

Disclosures

No relevant conflicts of interest to declare.
Antithymocyte Globulin-Based Prophylaxis for Graft Versus Host Disease Compared to Post-Transplant Cyclophosphamide-Based Prophylaxis in Matched Unrelated Donor Transplantation
Authors: Redondo Velao Sara , Kwon Mi , Champ Diana , Pascual Cascon MJ , Balsalobre Pascual , Diez J , Vidales Mancha I , Serrano David , Fernandez Noemi , Anguita Javier , Buño Ismael , Gayoso Jorge , Diez-Martin Jose Luis
Published: October 2019

INTRODUCTION

Post-transplant high-dose cyclophosphamide (PTCy) has shown to prevent graft versus host disease (GvHD) after haploidentical allogeneic hematopoietic cell transplantation (HSCT). Although less extended, its use after matched unrelated donor (MUD) HSCT in combination with additional immunosuppression has been encouraging. The aim of this study was to analyze and compare outcomes of antithymocyte globulin based (ATG) versus PTCy-based GvHD prophylaxis in matched unrelated donor HSCT.

MATERIAL AND METHODS

We retrospectively analyzed 51 MUD (8/8 HLA-matched) transplants performed in two Spanish centers: 25 received ATG-based prophylaxis and were performed from 2010 to 2013, and 26 received PTCy-based prophylaxis and were performed from 2013 to 2016.

RESULTS

Characteristics of patients and post-HSCT complications are detailed in Table 1. There were no significant differences in age, gender, diagnosis and Disease risk index. For the ATG group, conditioning regimen consisted of fludarabine combined with busulfan (80%) or melphalan (12%), and TBI plus cyclophosphamide (8%). All patients received ATG 2 mg/kg from day -4 to day -2 followed by methotrexate and cyclosporine (CsA). Conditioning regimen for patients from the PTCy group consisted of fludarabine and busulfan (85%) and thiotepa combined with busulfan and fludarabine (15%). GvHD prophylaxis consisted of PTCy 50 mg/kg on days +3 and +4, combined with: CsA plus MMF in 57%, CsA alone in 31%, or tacrolimus plus MMF or sirolimus in 8%, and tacrolimus alone in 4%.

With a median follow-up of 58 months for the ATG group and 12 months for the PTCy group, there were no statistically significant differences in overall survival at 18 months (56% vs 85%, p=0.08), in event free survival (56% vs 68%, p=0.5), and in cumulative incidence of relapse (12% vs 18%, p=0.3). Non-relapse mortality at 12 months was significantly higher in the ATG group (32% vs 10%, p = 0.04). The ATG group presented more cases of hepatic toxicity grades I-IV and hemorrhagic cystitis than PTCy group (Table 1). Cumulative incidence of acute GvHD grades II-IV was higher in the ATG group as well as acute GvHD grades III-IV, with no differences in moderate/severe chronic GvHD incidence (Figure A).

CONCLUSION

In our experience, albeit differences in follow-up and limited number of patients, we conclude that PTCy combined with additional immunosuppression after MUD allogeneic HSCT offers lower rates of acute GvHD together with less toxicity and non-relapse mortality compared to ATG-based prophylaxis. Further analysis including larger series and follow-up are needed to confirm these observations.

Disclosures

No relevant conflicts of interest to declare.
Hematopoeitic Stem Cell Transplant in Aplastic Anemia: Is It Time to Revise the Treatment Alogirthm
Authors: Gupta Ashish O. , Shein Steven L , Dalal Jignesh D
Published: October 2019

Background: Aplastic anemia (AA) is a rare bone marrow failure (BMF) syndrome of unknown etiology characterized by hypocellular bone marrow and severe persistent pancytopenia. Pediatric AA is more common in adolescent age group with better outcomes as compared to adults. Current treatment modalities include immunosuppressive therapy (IST) or matched related bone marrow transplant, when possible. There is a high risk of disease relapse with IST and clonal evolution into myelodysplastic syndrome. There is limited pediatric data on outcomes with different treatment options.

Methods: Retrospective database analysis of Pediatric Health Information Systems (PHIS) database was performed to evaluate healthcare outcomes of pediatric patients (age less than 21 years) with AA from January 1, 2006 to December 31, 2015. PHIS is an administrative quality-controlled database from 43 not-for-profit children's hospitals. ICD-9 code 284.9 was used to identify patients with acquired AA. Appropriate procedure and pharmacy billing codes were used to obtain information regarding transplants and various drugs used. SAS based statistical software was used to perform analysis.

Results: A total of 5127 inpatient admissions were noted for AA during the study period. An overall increasing trend of transplant was observed from 2006-2015 (Figure 1). Almost equal inpatient admission rate was noted in males and females, with a slightly higher rate amongst males who underwent transplant (55% vs 45%). Inpatient admission was more common amongst whites in both transplant (n=407, 8%) and non-transplant groups (n=4717, 92%; Table 1). Overall inpatient mortality was about 2% with a similar mortality in those who underwent transplant (5%). Cyclosporine (n= 1785, 38%) and steroids were most common immunosuppressive drugs used in non-transplant group. In the transplant group (Table 2), graft failure (20%) was the most common complication, while Cyclophosphamide (77%), Fludarabine (53%) and anti-thymocyte globulin (31%) based conditioning was most commonly used. Transplant itself was an independent risk factor for mortality (p=0.04) along with graft failure (p=0.02). Steroids (92%) and Cyclosporine (73%) were the most common immunosuppression used.

Conclusion: Hematopoietic stem cell transplants have good outcomes in pediatric AA patients. With an increasing trend of HSCT in these patients, successful outcomes with other transplant options such as match unrelated donor transplants should be considered. Treatment algorithm for Pediatric AA needs to be revised to include other transplant sources.

Disclosures

No relevant conflicts of interest to declare.
FVIII-Specific TCR Engineered Human Regulatory T Cells Suppress the Production of FVIII-Specific Responses Vitro and In Vivo
Authors: Kim Yong Chan , Zhang Aihong , Yoon Jeong-Heon , Scott David W.
Published: October 2019

Expanded antigen-specific engineered regulatory T cells (Tregs) have been proposed for potential clinical application for the treatment of undesirable immune responses, such as inhibitor responses in hemophilia A patients and autoimmune diseases. By providing an antigen-specific T-cell receptor (TCR) to polyclonal natural Tregs, we suggested that antigen-specific engineered Tregs would migrate specifically to particular target tissues and induce antigen-specific immune tolerance in the local milieu. Previously, we developed FVIII C2-specific Tregs using a long-term stabilization protocol in vitro and demonstrated that these stabilized engineered Tregs successfully modulated FVIII-specific T-cell and B-cell immune responses in vitro. Furthermore, these engineered Tregs could suppress T-effectors specific for additional epitopes in local milieu in both a cell contact and contactless manner. From these data, we hypothesized that IL-2 and related signaling pathways are major regulatory mechanisms of the suppression. To further investigate how IL-2R signaling is engaged to control T effectors and Tregs, we followed the phospho-STAT5 status of these cells kinetically. Our results showed clearly that IL-2 from activated T effectors is a key requirement for Treg activation, inducing subsequent blockage of STAT5 signal in T effectors by activated Tregs.

As further evidence of the efficacy of these specific Tregs, we then determined whether FVIII C2-specific Tregs could suppress the induction of FVIII inhibitor antibody in vivo. Thus, we transferred FVIII C2-specific human Tregs into HLA DR1 hemophilic mice and challenged them with FVIII in vivo. Our results showed that induction of FVIII-specific antibodies was inhibited for over 8 weeks. Taken together, our results suggest a potential therapeutic trial of FVIII-specific engineered Tregs in hemophilia A.

Disclosures

Kim: Henry Jackson Foundation: Patents & Royalties: Provisional submitted.
P27 Deficiency Results in Enhancing Hematopoiesis By Increasing IL22 Production from Bone Marrow Mesenchymal Stem Cells
Authors: Lu Rui-Nan , Wang Qian , Li Jianyong , Miao Dengshun
Published: October 2019

Hematopoietic microenvironment plays an important role in hematopoiesis, in which the bone marrow mesenchymal stem cells (BM-MSCs) can directly or indirectly affect the hematopoietic cells and regulate hematopoietic function. It has been reported that bone marrow hematopoietic progenitor cells were increased in p27 deficient mice, however, it is unclear whether BM-MSCs are mediated enhancing hematopoiesis resulting from p27 deficiency.

To answer this question, the conditioned medium (CM) were harvested from BM-MSC cultures derived from WT and p27 knockout (KO) mice, respectively and added to the cultures of bone marrow cells derived from WT mice. After 4 days, the sca-1+ckit+Lin- cells (hematopoietic stem cells, HSC) and sca-1+ckit+Lin+ cells (hematopoietic progenitor cells, HPC) in the resulting cells were analyzed by flow cytometry, and the assay for colony forming cells (CFCs) was performed and the colony numbers of the CFC were counted. Results showed that the HPC fraction and the number of CFCs were increased significantly, but the HSC fraction was not increased in the bone marrow cell cultures with the CM from p27 KO mice compared with those with the CM from WT mice.

To identify potential factors from the CM which enhance hematopoiesis caused by p27 deficiency, differences of protein expression profiles in the CM from the WT and p27 KO mice were examined by protein chip analysis. Five proteins were identified which were significantly increased in the CM from p27 KO mice, including interleukin-22 (IL-22), type I receptor of transforming growth factor-β, tumor necrosis factor- related apoptosis-inducing ligand, VE-cadherin and vascular endothelial growth factor B.

To determine whether IL22, as a paracrine factor, plays a role in stimulating hematopoiesis by activating JAK-STAT signal pathway mediated by IL22 receptor A1 (IL22RA1), we examined the effect of IL22 on HPCs and the expression of signal molecules of JAK-STAT pathway. Results showed that the expression levels of IL22 at both mRNA and protein levels were up-regulated significantly in BM-MSCs derived from p27 deficient mice compared with those from WT mice. The HPC fraction and the number of CFCs were increased significantly in bone marrow cultures in the present of IL22, and were decreased significantly in the cultures with the CM from p27 knockout mice containing IL22 antibody. The percentages of IL22RA1, STAT3 and p-STAT3-S727 positive HPCs were increased significantly in the bone marrow from p27 KO mice compared with that from WT mice.

Results in this study indicate that IL22, as a paracrine factor, might play an important role in stimulating hematopoiesis by activating JAK-STAT signal pathway mediated by IL22RA1.

Disclosures

No relevant conflicts of interest to declare.