bluuebunny/crossref_metadata_2025 huggingface dataset:
Components of the Metabolic Syndrome in 500 Adult Long-Term Survivors of Childhood Cancer.
Authors: van Waas Marjolein , Neggers Sebastian , Pieters Rob , Van Den Heuvel M.M.
Published: October 2019

Abstract 3068

Poster Board III-5

Introduction

Adult survivors of childhood cancer have been reported to have an increased risk of late sequels. A cluster of abnormalities that contribute to the metabolic syndrome may be expressed at a higher level and therefore result in an increased risk for diabetes mellitus and cardiovascular diseases.

Patients and Methods

We investigated a single centre cohort of 500 adult survivors (228 females) of childhood cancer, median age 28 years (range 18-59 years) and median follow-up time 19 years (range 6–49 years). This cohort included 164 acute lymphoblastic leukaemia (ALL) survivors (75 females). We measured total cholesterol, high-density lipoprotein-cholesterol (HDL), systolic and diastolic blood pressure, body mass index and the prevalence of diabetes mellitus. Data from the Dutch epidemiologic MORGEN-study were used to calculate standard deviation scores as normative values.

Results

The criteria of the metabolic syndrome were met in 13% of the total cohort. ALL survivors treated with cranial irradiation had an increased risk of developing the metabolic syndrome compared to ALL survivors not treated with cranial irradiation (23% vs. 7%, P=0.011). ALL survivors who received CRT had higher total cholesterol levels compared to ALL survivors who did not (mean SDS 0.38 vs. mean SDS –0.05, P=0.027), whereas their HDL levels did not differ. Also, ALL survivors treated with CRT were more often hypertensive compared to ALL survivors not treated with CRT (22% vs. 10%, P=0.036) and more often overweight (59% vs. 34%, P=0.003), however they were not more often obese (12% vs. 9%, ns).

Conclusions

Adult survivors of childhood cancer, especially ALL survivors treated with cranial irradiation, are at increased risk of developing the metabolic syndrome. This increased risk is probably determined by higher prevalence of overweight and hypertension in ALL survivors.

Disclosures

No relevant conflicts of interest to declare.
Pharmacogenetic Risk Factors for Altered Bone Mineral Density and Body Composition in Pediatric Acute Lymphoblastic Leukemia.
Authors: te Winkel Mariël L. , van Beek Robert D , de Muinck Keizer-Schrama Sabine M.P.F. , Uitterlinden André G. , Hop Wim C.J. , Pieters Rob , Van Den Heuvel M.M.
Published: October 2019

Abstract 3071

Poster Board III-8

Introduction

As the survival of pediatric acute lymphoblastic leukemia (ALL) improves, research on treatment-related morbidity, like disturbance of body composition and bone mineral density (BMD), is required. This study investigates pharmacogenetic risk factors for BMD and body composition in pediatric ALL.

Methods

We determined the influence of SNPs in 4 genes (vitamin-D receptor (VDR: BsmI/ ApaI/ TaqI and Cdx-2/ GATA), collagen type I alpha 1 (SpI), estrogen receptor 1 (ESR1: PvuII/ XbaI) and the glucocorticoid receptor (BclI)) on body composition, BMD and fracture risk during dexamethasone-based pediatric ALL treatment. Anthropometry data of 69 patients (mean age 7.4 (range: 1.6-16.8) year) treated according to the DCOG-ALL9 protocol were evaluated. In patients aged >4 years body composition and BMD of the total body (BMD-TB) and the lumbar spine (BMD-LS) were measured repeatedly using dual energy X-ray absorptiometry. To correct for bone size we calculated bone mineral apparent density of the lumbar spine (BMAD-LS) with the model BMAD-LS = BMD-LS x (4/ (n × width)). All values were expressed as standard deviation scores (SDS). Repeated measurements analysis (ANOVA) was used.

Results

Non-carriers of VDR 5'-end (Cdx-2/ GATA) haplotype 3 revealed a significant larger fat gain than carriers (Δ%fat: non-carriers: +1.76 SDS, carriers: +0.77 SDS, p<0.001). At diagnosis and during therapy, BMD-LS was significant higher in non-carriers of VDR 5'-end (Cdx-2/ GATA) haplotype 3 than in carriers. The other SNPs did not influence BMD or fracture risk during or after treatment. The year after treatment discontinuation, lean body mass increased in non-carriers of ESR1 (PvuII/ XbaI) haplotype 3 and decreased in carriers (Δ lean body mass: non-carriers: +0.28 SDS, carriers: -0.55 SDS, p<0.01).

Conclusion

This is the first study investigating the influence of genetic variation of the VDR, COLIA1, ESR1 and GR on body composition, BMD and fracture risk in pediatric ALL. We found the VDR 5'-end (Cdx-2/GATA) haplotype 3 as a protective factor for excessive fat gain during therapy. Moreover, this haplotype 3 of the VDR 5'-promoter was determined as risk factor for a lower BM(A)D-LS at diagnosis that remained during ALL treatment. Carriage of ESR1 (PvuII/XbaI) haplotype 3 negatively influenced recovery of lean body mass after treatment discontinuation.

Disclosures

No relevant conflicts of interest to declare.
Prognosis of Secondary Acute Myeloid Leukemia Is Affected by the Type of the Preceding Hematologic Disorders and the Presence of Trisomy 8.
Authors: Koh Youngil , Kim Inho , Kim Hyun-Kyung , Yoon Sung-Soo , Lee Dong Soon , Park Seonyang , Kim Byoung Kook
Published: October 2019

Abstract 3117

Poster Board III-54

Differences in the clinical course of secondary acute myeloid leukemia (s-AML) according to the type of the preceding disorders are not defined. We compared the outcomes of therapy-related AML (t-AML), AML following myelodysplastic syndrome (MDS-AML) and AML following myeloproliferative disorder (MPD-AML). We also intended to find prognostic factors in s-AML overall.

Retrospective medical record review at Seoul National University Hospital was performed. We assessed response to induction chemotherapy and overall survival (OS).

95 s-AML patients (median age 56.4 years) were analyzed. 26, 57 and 12 patients had t-AML, MDS-AML and MPD-AML respectively. For patients receiving induction chemotherapy, complete remission (CR) rate was 47.5% and CR rate was different according to the type of the preceding disorders (p=0.004). Compared to t-AML (p=0.027) and MDS-AML (p=0.050), MPD-AML had shorter OS. In s-AML, presence of trisomy 8 had a prognostic impact (p=0.003) along with cytogenetic risk group (p=0.016). In multivariate analysis, the type of the preceding disorders (p=0.026), 5q deletion (p=0.015) and trisomy 8 (p=0.040) were independent prognostic factors.

In conclusion, prognosis of s-AML was different according to the type of the preceding disorders with the worst prognosis in MPD-AML. Along with cytogenetic risk grouping, trisomy 8 had a prognostic impact in s-AML.

Table 1 Characteristics of s-AML patients according to preceding disorders Characteristics t-AML AML-MDS AML-MPD p-value Age (median) 54.1 57.2 65.5 0.888 Gender 0.262 Male 13 39 8 Female 13 18 4 Latency period 31.2 6.7 92.6 0.008 White blood cell (median) (/μL) 5615 2905 12785 0.002 Hemoglobin (median) (g/dL) 8.0 8.4 8.7 0.543 Platelet (median) (/μL) 51500 32000 100500 0.509 Risk group 0.001 Favorable 6 0 0 Intermediate 12 37 10 Poor 5 12 2 5q deletion 0.731 Present 5 7 2 Absent 18 42 10 -7/7q deletion 0.107 Present 1 11 1 Absent 22 38 11 Trisomy 8 0.777 Present 6 13 2 Absent 18 36 10 Induction chemotherapy 0.619 Yes 23 47 11 No 3 10 1 Complete remission after induction 0.004 Yes 17 19 2 No 6 27 9 Stem cell transplantation 0.137 Yes 9 4 1 No 8 15 2

Figure 1 CR-1 (A) and OS (B) according to preceding disorders CR-1 was not affected by preceding disorders, however OS was significantly the shorter in AML evolving from MPD (black line) compared to AML evolving from MDS (orange line) and t-AML (green line). Figure 1. CR-1 (A) and OS (B) according to preceding disorders CR-1 was not affected by preceding disorders, however OS was significantly the shorter in AML evolving from MPD (black line) compared to AML evolving from MDS (orange line) and t-AML (green line).

Disclosures

No relevant conflicts of interest to declare.
Atorvastatin Reduces Collagen-Induced Platelet Function in Aspirin-Treated Patients with Acute Myocardial Infarction: Mechanisms Involved.
Authors: Teresa Santos Maria , Fuset Maria Paz , Ruano Miguel , Moscardo Antonio , Cortina Belen , Dasi Maria Angeles , Valles Juana
Published: October 2019

Abstract 3126

Poster Board III-63

Treatment with statin at the onset of acute myocardial infarction (AMI) has demonstrated a clinical benefit. The effects of statins on platelet TXA2 synthesis or platelet function could be involved in the clinical benefit, but has not been previously evaluated in patients with AMI. Collagen-induced (1μg/mL)-platelet aggregation, activation (14C-5HT release, TXA2 synthesis) and platelet recruitment (proaggregatory activity of activated platelets cell-free releasate) (1, 2) were evaluated in 184 patients with AMI within 48h of the onset of the acute event. Patients were divided into 2 groups: treated with aspirin (ASA) alone (n=139) and treated with both ASA and atorvastatin (AT; n=45). Insufficient inhibition of TXA2 (<95% vs. ASA-free normal subjects) was detected in 25% of patients treated with ASA vs. 9% in the group treated with ASA+AT (p<0.05), and persistent TXA2 synthesis was reduced by 87.5% in patients treated with ASA+AT vs. ASA alone. Reduction of TXA2 by AT was associated with significant inhibitions of collagen-induced: aggregation (30%), 14C-5HT release (39%) and recruitment (33%). To evaluate the biochemical mechanisms involved in the AT effect on TXA2 synthesis, experiments in vitro with washed platelets from normal subjects were performed. Incubation of AT alone (1-20μM) or AT plus a suboptimal ASA concentration (1μM) resulted in a significant inhibition of collagen-induced (1μg/mL) TXA2 synthesis and aggregation, confirming the results in the patients. In contrast, AT did not influence arachidonic acid-induced aggregation, suggesting an effect of the statin on the cytosolic phospholipase A2 (cPLA2) activity rather than on cyclooxygenase-1. cPLA2 activity is known to be regulated by its phosphorylation by MAPK (p38 or erk 1/2) and the increase in cytosolic Ca2+. In kinetic studies we found that AT reduced collagen-induced cPLA2 and erk 1/2 phosphorylation, but did not modify p38 phosphorylation. In addition, AT almost blocked collagen-induced increase in cytosolic Ca2+. In conclusion, AT associated with ASA significantly reduced collagen-induced persistent TXA2 synthesis and platelet reactivity early at the onset of AMI, reducing aspirin resistance in these patients. This effect is mediated by the AT control of cPLA2 activity via both erk1/2-mediated cPLA2 phosphorylation and blockade of collagen-induced elevation of cytosolic calcium. 1. Santos MT et.al. J Clin Invest 1991;87:571-80; 2. Vallés J et.al. Blood 2002;99:3978-84.FIS07/0463/MMA2006/RD06/0026

Disclosures

No relevant conflicts of interest to declare.
Age of Red Blood Cell Units Transfused in Canada.
Authors: Kokolo Madzouka , Tinmouth Alan , Fergusson Dean , Giulivi Antonio , Hebert Paul
Published: October 2019

Abstract 3143

Poster Board III-80

Background

During storage, red blood cells (RBCs) undergo well-described biochemical and biomechanical changes, termed the red cell storage lesion. These changes may affect the ability of stored RBCs to deliver oxygen or have other adverse consequences. The transfusion of older RBCs has been associated with increased morbidity/mortality, and planned or on-going randomized controlled trials are exploring a possible causal relationship between the age of RBCs and mortality/morbidity. However, the age-related characteristics of the RBCs units in hospital inventories and of the RBCs units transfused are largely unknown.

Method

A descriptive observational study from 9 centres from 5 Canadian provinces was conducted. Using a randomly generated schedule, data was collected on one weekday for 30 consecutive weeks. Reports of the RBCs inventories and the transfused RBCs units for all ABO and Rh blood groups were collected and analyzed.

Results

Academic centers contributed more than half of the collected data, both for inventory and for transfused units. Overall, the median age of RBCs units in inventory was 14 days (IQR=10-20) and their median age upon transfusion was 16 days (IQR=12-22). The cumulative proportion of younger blood (less than 8 days) in inventory was 9% (range for individual centres: 5 to 20%) for units in inventory and 5% (range for individual centres: 1 to 17%) for transfused units. The cumulative proportion of older blood (more than 28 days) was 8% (range for individual centres: 6 to 14%) for units in inventory, and 13% (range for individual centres: 9 to 23%) for transfused units. Community centres tended to have the largest proportion of younger units transfused (12%) as well as in inventory (14%).

Conclusion

This study provides important data that may contribute to decision-making regarding blood bank inventory management and age-based transfusion practices. If the results of ongoing studies demonstrate an adverse effect from transfusing older RBCs, then significant changes to the inventory management may be required. In Canada, shortening the shelf-life for storage of RBC units to 28 days may be less problematic, but restricting the age of RBCs transfused to less than 8 days for all RBC units may pose significant challenges.

Disclosures

Tinmouth: Canadian Blood Services: Consultancy; Novartis: Research Funding; CSL: Research Funding; Bayer: Research Funding; Wyeth: Research Funding.
Exploratory in Vitro Study of Red Blood Cell Storage Container with An Alternative Plasticizer.
Authors: Dumont Larry J. , Baker Sharry , Waters Susan L , Herschel Louise , Dumont Deborah F , David Raymond M , Otter Rainer
Published: October 2019

Abstract 3149

Poster Board III-86

Background

The plasticizer di-2-ethylhexyl phthalate (DEHP) is a common component in medical plastics. Unique to the storage of RBC, DEHP is a necessary component of these storage bags, and is required to prevent excessive hemolysis over storage. Due to concerns related to the long term effects of DEHP exposure and issues related to disposal of these materials, there is a drive to replace this component. Our objective is to evaluate a candidate replacement plasticizer (Hexamoll TM DINCH, Cyclohexane-1,2-dicarboxylic acid, dinonylester, branched and linear) compared to control DEHP in an in vitro feasibility study. We hypothesize that the candidate will provide at least equivalent protection against hemolysis for RBC stored in additive solution for 42 days, and periodic mixing of RBC stored in additive solution for 42 days will add additional protection against hemolysis.

Methods

Whole blood was collected into CPD in standard collection sets (Terumo), combined into pools of 2 ABO identical RBC, divided, leukocyte-reduced, centrifuged and separated into plasma and packed RBC, then additive solution (AS-5) was added to the RBC, and the RBC/AS-5 were transferred into 2 study bags (matched pair) for storage up to 42 days under standard blood bank conditions of 4°C. In the first part, both study bags were Hexamoll TM DINCH, and one bag of each pair was mixed once per week, the other was held undisturbed. In the second part, DEHP bags were used as control held 42 days undisturbed, and the DINCH test bag was mixed once per week during storage. Standard in vitro RBC characteristics were determined at days 0 and 42.

Results

DINCH bags mixed weekly retained RBC morphology better, had lower extracellular potassium and had lower hemolysis compared to static storage. There were no detectable differences between DEHP static stored bags and DINCH bags mixed weekly when comparing in vitro parameters.

Conclusion

Periodic mixing of RBC stored in DINCH bags provides additional protection against hemolysis over 42 days of storage. DINCH plasticizer provided at least equivalent protection against hemolysis and potassium leakage for RBC stored in additive solution for 42 days. These initial observations should be extended to larger sample sizes and to in vivo autologous RBC recovery studies. The need for periodic mixing will need to be evaluated further. The plasticizer DINCH may be a viable candidate for replacement DEHP in RBC storage bags.

Disclosures

Dumont: BASF: Research Funding. Baker:BASF: Research Funding. Waters:BASF: Research Funding. Herschel:BASF: Research Funding. Dumont:BASF: Research Funding. David:BASF: Employment. Otter:BASF: Employment.
E3 Ligase c-CBL Mediates Ubiquitination-Proteasomal Degradation of BCR-ABL and Therapeutic Effects against BCR-ABL Leukemia.
Authors: Mao Jianhua , Sun Xiaoyan , Zhang Qun-Ye , Huang Qiu-Hua , Liu Ping , Xie Yinyin , Lu Jing , Chen Saijuan , Chen Zhu
Published: October 2019

Abstract 3257

Poster Board III-1

We previously demonstrated that the cellular level of the fusion oncoprotein BCR-ABL in chronic myelogenous leukemia (CML) could be regulated by the proteolysis via ubiquitin-proteasome pathway and arsenic sulfide (As4S4) could enhance this process to exert therapeutic effects against CML. However, the exact biochemical mechanism and molecules involved, including the ubiquitin E3 ligases of BCR-ABL, their modification sites and the topologic type of ubiquitination remain uncovered. In this work, by using immunoprecipitation (IP) /2D-nano-HPLC/MALDI-TOF-TOF, we found a number of components in ubiquitination process, such as ubiquitin, E1, E2 and E3 ligases in the BCR-ABL protein complex. Particularly, several E3 ligases including the CBL family E3 ligase c-CBL were identified. The in vitro ubiquitination experiments indicated that c-CBL was one of the E3 ligases of BCR-ABL. Overexpression and knockdown experiments of the c-CBL gene revealed it participated in the process of endogeneous BCR-ABL ubiquitination and turnover. The enforced overexpression of c-CBL could enhance the degradation of BCR-ABL and apoptosis of CML cells whereas the down regulation of c-CBL generated opposite effects. Furthermore, we mapped a domain of c-CBL as the interface to form complex with BCR-ABL based on the protein structure prediction. Either the identification of BCR-ABL ubiquitination sites or the interaction experiment among the c-CBL and BCR-ABL are undergoing. Our data thus suggest that induction of c-CBL may be a new therapeutic approach for CML.

Disclosures:

No relevant conflicts of interest to declare.
Loss of Growth Arrest DNA Damage 45a,b (GADD45a,b) Enhances Oncogenicity in BCR/ABL-Driven Chronic Myelogenous Leukemia.
Authors: Sha Xiaojin , Liebermann Dan A. , Hoffman Barbara
Published: October 2019

Abstract 3264

Poster Board III-1

The bcr/abl oncogene causes chronic myelogenous leukemia (CML) in human. BCR/ABL induces the transformation of myeloid lineage through MAPK, JNK/SAPK, PI3K signaling pathways. Growth arrest DNA damage 45A (GADD45A) and GADD45B are upregulated during myeloid lineage terminal differentiation. They are involved in G2/M cell cycle arrest and apoptosis in response to exogenous stress stimuli through MAPK and JNK/SAPK pathways. To investigate the effect of GADD45A and GADD45B in the development of CML, syngeneic wild type lethally irradiated mice were reconstituted with wild type, gadd45a or gadd45b null myeloid progenitors transduced with a retrovirally expressed 210-kD BCR/ABL fusion oncoprotein. We found that loss of gadd45a or gadd45b accelerated the development of CML-like disease in wild type recipients. BCR/ABL transformed gadd45a or gadd45b deficient progenitor recipients exhibited a significantly accelerated kinetics of increase in the number of WBC and percentage of myeloid blasts in blood compared to mice reconstituted with the same number of wild type bone marrow cells transduced with BCR/ABL. There was also increase in the rate of accumulation of CD11b+Gr1+ cells in the bone marrow and spleen. Using in vitro and in vivo BrdU assays, enhanced proliferation capacity was observed for both BCR/ABL transduced gadd45a and gadd45b deficient myeloid progenitors. BCR/ABL transduced gadd45a and gadd45b deficient primary myeloid progenitors formed more and bigger colonies compared to BCR/ABL transformed wild type progenitors. Impaired apoptosis was showed in BCR/ABL transduced gadd45a deficient myeloid progenitors. These results indicate that both gadd45a and gadd45b function as suppressors of the development of BCR/ABL driven CML, where gadd45a appears to suppress CML via mechanism involving inhibition of cell proliferation enhancement of apoptosis, whereas gadd45b appears to only inhibit cellular proliferation. Dissecting the molecular nature of signaling paths involved in the suppressive function of gadd45a and gadd45b in BCR/ABL driven CML, as well as analysis of Gadd45 in CML patients, is underway.

Disclosures:

No relevant conflicts of interest to declare.
Sub-Morphologic Evidence of Disease Prior to Stem Cell Transplantation Correlates with Inferior Post Transplant Outcome in Childhood Acute Myeloid Leukemia.
Authors: Rosenberg Abby R. , Leisenring Wendy , Sanders Jean E , Woolfrey Ann E , Carpenter Paul A. , Wells Denise A. , Loken Michael R. , Meshinchi Soheil
Published: October 2019

Abstract 328

Hematopoietic cell transplantation (HCT) is utilized in the treatment of a subset of children with acute myeloid leukemia (AML). Although presence of minimal residual disease (MRD) has been associated with a high relapse rate in children with AML, the clinical implications of MRD prior to HCT have not previously been defined. We explored the clinical outcomes associated with the presence of morphologic or sub-morphologic disease prior to HCT. In the period of 1995-2007, 89 patients age 1-18 years received myeloablative conditioning and allogeneic HCT from related (N=36) or unrelated (N=53) donors at Fred Hutchinson Cancer Research Center. At the time of HCT, 63 patients received transplants in complete remission (CR) and 26 had morphologic disease. Among those in morphologic CR, 43 were in 1st CR, 17 in 2nd CR and 3 were in 3rd CR. All patients had bone marrow evaluation for MRD by multi-dimensional flow cytometry (MDF) 2-4 weeks prior to HCT. MDF testing showed that 10 of 63 patients (16%) who received transplants in morphologic CR had MRD levels ranging from 0.3% to 12% (median 3.7%) while the remaining 53 patients had no detectable MRD. The median follow up for the 38 survivors is 6 years (range 1 – 12). Actuarial overall survival at 3 years from HCT for patients who received HCT with and without morphologic disease was 12% vs. 59%, (p<0.001) with corresponding relapse rates of 62% and 24%, (p<0.001), respectively. We subsequently evaluated clinical outcomes for patients who received HCT in morphologic CR with and without evidence of MRD by MDF. Actuarial overall survival for patients who received transplants in morphologic CR with MRD was 15% vs. 67% for those without MRD, p=0.001. The corresponding relapse rate for those with MRD was 65% vs. 17% for those without MRD (p<0.001). Among the 10 CR patients with MRD, 4 had high risk cytogenetics and 6 patients had standard risk cytogenetics. In 53 MRD-negative patients who received HCT in CR, 10 (19%) had high risk, 7 had favorable risk cytogenetics and the remaining 37 (69%) had standard risk cytogenetics. It is notable that the relapse and overall survival probabilities for patients who were received HCT in morphologic CR but with MRD appear similar to those for subjects in morphologic relapse. This data demonstrates that presence of sub-morphologic disease prior to HCT portends high relapse risk and worse post transplant survival. Larger studies are evaluating the full impact of pre-transplant MRD as well and the clinically significant threshold of MRD prior to stem cell transplantation in AML.

Disclosures:

No relevant conflicts of interest to declare.
Hematopoietic Cell Transplantation for Chronic Lymphocytic Leukemia: Similar Efficacy of Non-Myeloablative and Myeloablative Conditioning; Long-Term Evaluation From the University of Minnesota.
Authors: McClune Brian , Burns Linda J. , DeFor Todd E. , Slungaard Arne , Bachanova Veronika , Brunstein Claudio , Weisdorf Daniel J.
Published: October 2019

Abstract 3385

Poster Board III-273

Allogeneic hematopoietic cell transplantation (HCT) for chronic lymphocytic leukemia (CLL) is increasingly utilized. We retrospectively analyzed outcomes of HCT for 43 patients with CLL at the University of Minnesota from February 1985 through March 2009 using myeloablative (MA, n=19) (total body irradiation [TBI] 1320 cGY in 8 fractions and cyclophosphamide [Cy] 60 mg/kg x 2 days) and non-myeloablative (NMA, n=24) preparative regimens (fludarabine 40 mg/m2 × 5 days or cladribine 50 mg/m2 IV with TBI 200cGy in 1 fraction and either oral busulfan [Bu] 8 mg/kg divided every 6 hours over 2 days or Cy 50 mg/kg x1 day). Graft-versus-host disease (GVHD) prophylaxis included cyclosporine (CSA) and methotrexate in most MA patients and CSA and mycophenolate mofetil (MMF) for all NMA patients. Median follow up for survivors was 3.3 years. Nine-two percent of NMA HCT took place after 2000 whereas only 26% of MA HCTs happened during this time. Most patients were male (15/19 MA, 20/24 NMA). Time from diagnosis to treatment was nearly twice as long in NMA vs. MA patients (median 72 vs. 39 months) with MA patients progressing through a median of 3 regimens (range, 1-5) and NMA 4 (1-7). Poor-risk cytogenetics (11q-, 17p-) were found in 5 MA and 10 NMA patients. Seven and 13 patients were fludarabine refractory in the MA and NMA groups, respectively. Eighteen patients in the MA group had matched related donors (MRD), while only 10 MRD were amongst the NMA patients. Umbilical cord blood stem cells were used in 12 NMA patients. Neutrophil recovery was slightly faster after NMA HCT vs. MA (median 12.5 vs. 18.5 days; p=0.14). Incidence of acute GVHD, grades II-IV, was similar (p=0.11) but grades III-IV at day 100 was more frequent in the NMA than the MA group (33% vs. 5%, p=0.04). The chronic GVHD incidence was 43% (95% CI, 18-68%) in MA and 29% in NMA (11-47%, p=0.08) patients. Transplant-related mortality (TRM) at 1 year was 42% (20-64) in the MA arm vs. 25% (8-42) (p=0.34) for the NMA group. Relapse rates at 3 years were higher in the NMA HCT group (MA 18% [0-36] vs. NMA 26% [8-44]). Three-year progression-free survival (PFS) was similar in MA (33% [13-55]) and NMA (39% [19-58]) patients (p=0.61). Overall survival at three years was also similar (MA 37%, [15-60] vs. NMA 52%, [30-70]) (p=0.28). While these data suggest that dose intensity may affect TRM and relapse, NMA HCT yields equivalent and encouraging PFS and OS as compared to MA HCT. Our results support further investigation of the role of allogeneic transplantation in the treatment of high risk and advanced CLL.

Disclosures:

No relevant conflicts of interest to declare.
Nonmyeloablative Allogeneic HCT From Unrelated Donors for Multiple Myeloma (MM).
Authors: Rotta Marcello , Storer Barry E , Lange Thoralf , Pulsipher Michael , Shizuru Judith , Bruno Benedetto , McSweeney Peter A. , Agura Edward , Chauncey Thomas , Appelbaum Frederick R , Bensinger William , Sandmaier Brenda M , Storb Rainer F , Georges George E. , Maloney David G
Published: October 2019

Abstract 3391

Poster Board III-279

BACKGROUND:

We previously reported on the outcome of unrelated donor nonmyeloablative hematopoietic cell transplantation (HCT) in 24 patients (pts) with poor-risk multiple myeloma treated by Seattle Consortium Centers (Georges et al BBMT 2007). Here we update our observation to 43 pts with a median follow up of 3.3 years after allografting.

PATIENTS:

Pts with stage II-III MM (n=43) received AlloHCT at 9 centers between May 2000 and September 2008. Forty pts (93%) were matched with their donors for 10 of 10 HLA alleles, and 3 (7%) had single HLA-C allele-level mismatches. Median age at allotransplant was 53 (range 35–67) years. Median number of prior treatments was 2 (1–3), and median number of prior treatment cycles was 8 (5–22). All pts but 2 received at least 1 (range 1-3) high dose-Autograft regimen. Fifteen pts (35%) received planned tandem Auto/AlloHCT as consolidation to first line therapy. Allogeneic conditioning was with 2 Gy TBI plus fludarabine 90 mg/m2 and post allografting immunosuppression was with mycophenolate mofetil (MMF) and cyclosporine or tacrolimus. Disease status at allogeneic HCT included complete remission (CR, 6 pts, 14%), very good partial remission (VGPR, 12 pts, 28%), partial remission (PR, 14 pts, 33%) and refractory disease (RD, 11 pts, 26%).

RESULTS:

All pts had sustained donor engraftment. Twenty-eight (65%) developed grade 2 to 4 acute graft-versus-host-disease (GVHD) and 6 pts (14%) developed 3 to 4 acute GVHD. Twenty-six pts (60%) had extensive chronic GVHD. The overall response rate was 86%, with 18 pts (42%) achieving CR, 14 (33%) VGPR and 5 (12%) PR. With a median follow-up of 3.3 (0.3–8.1) years from allografting, median time to progression was 1.1 years. Median overall survival (OS) has not been reached. Median progression-free survival (PFS) was 1.5 years. Five-year estimated OS and PFS were 51% and 21% respectively. Cumulative incidence of nonrelapse mortality (NRM) at 100 days, 1 and 5 years were 2%, 16% and 19% respectively. The subgroup of 15 pts receiving upfront tandem Auto/AlloHCT had five-year estimated OS and PFS of 72% and 37%, respectively. These results are similar to the outcomes we observed in a series of 102 patients with MM who received upfront tandem Auto/AlloHCT from HLA-identical sibling donor (Rotta et al, Blood 2009) where five-year OS and PFS were 64% and 36%, respectively.

CONCLUSION:

The use of unrelated donors leads to sustained donor engraftment and is associated with a low 1-year NRM (16%). As consolidation of first remission, Tandem Auto/Allo HCT leads to similar 5-year outcomes as HCT from HLA-identical sibling donors.

Disclosures:

No relevant conflicts of interest to declare.
Outcome of HLA-Mismatched Unrelated Donor Allogeneic Stem Cell Transplantation Depends On the Intensity of the Conditioning Regimen.
Authors: Ayuk Francis , Zabelina Tatjana , Bacher Ulrike , Wolschke Christine , Lellek Heinrich , Eiermann Thomas , Erttmann Rudolf , Kröger Nicolaus , Zander Axel R.
Published: October 2019

Abstract 3393

Poster Board III-281

Background:

HLA-mismatched unrelated donors are increasingly becoming a graft source for both standard myeloablative (MAC) and reduced intensity conditioning (RIC) hematopoetic stem cell transplantation (HSCT).

Patients and methods:

We retrospectively analysed the effects of HLA-mismatching in 553 patients undergoing MAC (n = 342) or RIC (n = 211) unrelated donor HLA-matched (n = 289) or -mismatched (n = 264) HSCT with antithymocyte globulin as part of conditioning. Patient characteristics well matched between HLA-matched and –mismatched groups of MAC and RIC patients.

Results:

Median follow-up was 1946 days for MAC patients and 765 days for RIC patients. In MAC patients, there was no difference in the incidence of aGvHD, treatment related mortality (TRM) and overall survival (OS) between recipients of HLA-matched vs. –mismatched allografts. In RIC patients on the contrary, the incidence of aGvHD II-IV (46%, vs. 32.0% p = 0.05), III-IV (18%, % vs. 9.0 p = 0.07) and TRM (35% vs. 20%, p = 0.04) were higher and OS lower (31% vs. 50%, p = 0.001) for recipients of HLA-mismatched vs. –matched transplants. In the RIC patients, female donor gender (RR: 1.8; p = 0.02) and HLA-mismatch (RR: 1.8; p = 0.02) negatively influenced TRM while female donor gender (RR: 1.58, p = 0.03), HLA-mismatch (RR: 1.82, p = 0.003) and bad risk disease (RR: 2.14, p < 0.001) negatively impacted OS.

Conclusion:

Our analyses surprisingly reveal that the positive effect of ATG in HLA-mismatched transplants is only limited to patients undergoing standard myeloablative conditioning. HLA-mismatching and female donor gender negatively impact outcome of RIC allogeneic unrelated donor HSCT despite the use of ATG, highlighting the need for improved strategies.

Acknowledgments

We thank the staff of the BMT unit for providing outstanding care to our patients and the medical technicians for their excellent work in the BMT laboratory.

Disclosures:

Off Label Use: Antithymocyte globulin for prevention of severe GVHD.
Effect of the Deacetylase Inhibitor Suberoylanilide Hydroxamic Acid (SAHA) On Chronic Lymphocytic Leukemia Cells: Apoptosis, Migration Impairment and Drug Sensitization.
Authors: Stamatopoulos Basile , Meuleman Nathalie , De Bruyn Cécile , Delforge Alain , Bron Dominique , Lagneaux Laurence
Published: October 2019

Abstract 3435

Poster Board III-323

Chronic lymphocytic leukemia (CLL) is a neoplastic disorder that arises largely as a result of defective apoptosis leading to chemoresistance. Furthermore, SDF-1 and its receptor CXCR4 has been shown to play an important role in CLL cell trafficking and survival. Since histone acetylation is involved in the modulation of gene expression, cellular differentiation, and survival, we evaluated the effects of suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, on CLL cells in vitro and in particular on cell survival, CXCR4 expression, migration and in combination with different chemotherapies. Here, we showed that a 48-hour treatment of SAHA induced a dose-dependent decrease in CLL cell viability via apoptosis (n=20, p=0.0032). This effect was also seen in previously untreated and chemoresistant CLL patients. Using specific caspase inhibitors, we demonstrated the participation of caspases-3, -6 and -8, suggesting an activation of the extrinsic pathway. Additionally, SAHA decreased actin polymerization (about 45%) in response to SDF-1 (n=6, p=0.0313). SAHA also significantly decreased CXCR4 mRNA (n=10, p=0.0010) and protein expression (n=25, p<0.0001). Consequently, CLL cell migration in response to SDF-1 (n=17, p=0.0003) or through bone marrow stromal cells (pseudoemperipolesis) was dramatically impaired. Finally, SAHA at low concentration (5μM) could increase sensitivity of CLL cells to fludarabine, flavopiridol, thalidomide or bortezomib. In conclusion, SAHA induces apoptosis in CLL cells via the extrinsic pathway and downregulates CXCR4 expression leading to decreased cell migration. SAHA (alone or in combination with other drugs) represents thus a promising therapeutic approach to inhibiting migration, inducing apoptosis in CLL cell and potentially overcoming drug resistance.

Disclosures

No relevant conflicts of interest to declare.
Downregulation of Mir-22 in ALL Cells Is Associated with the Accumulation of Histone Modification but Independent of DNA Methylation.
Authors: Li Xiaoqing , Liu Jun , Zhou Rui , Huang Shi , Chen Xian-Ming , Huang Shiang
Published: October 2019

Abstract 3464

Poster Board III-352

MicroRNA-22 is one of the miRNAs frequently downregulated in human ALL cells and may play an important anti-tumor role in normal hematopoiesis. Histone modification and DNA methylation can have different roles in gene silencing in cancer. To investigate whether histone modifications would contribute to the dysregulation of miRNA-22 in acute lymphoblastic leukemia (ALL), the effect of a histone deacetylase inhibitor, trichostatin A (TSA), on miRNA-22 expression of primary ALL cells was analyzed by real-time PCR. The total number of patients included to this study is 33, including 26 samples of leukemia (18 of ALL and 8 of acute myeloid leukemia) and 7 normal controls. All patient blood samples were collected at the time of diagnosis. We detected a lower expression of pri-miR-22 in PMBCs from ALL patients compared with that from the health volunteers. Treatment with TSA significantly increased pri-miR-22 expression in PMBCs from ALL patients, but not in cells from the health volunteers. Whereas PMBCs from ALL patients and AML patients showed comparable levels of pri-miR-22. TSA treatment had no effect on pri-miR-22 expression in PMBCs from AML patients, suggesting TSA-mediated upregulation of miR-22 transcription in ALL but not AML malignant cells. Moreover, we used MPS assay to analyze the methylation status at the promoter element of miR-22 gene in primary human specimens. No DNA hypermethylation was detected in PMBCs from the health volunteers and patients with either ALL or AML. These data provide further evidence that miR-22 silencing in ALL cells may be DNA methylation-independent. In contrast, accumulation of the repressive histone marker H3K27 trimethylation (H3K27triM) was indentified around the transcriptional start point of the gene, which reduced by TSA treatment.

In conclusion, we showed that histone modification is involved in miRNA dysregulation in human ALL cells. Specifically, the silencing of miR-22 in ALL cells is associated with the accumulation of histone modification in its promoter element of miR-22 gene but independent of DNA methylation. The accumulation of H3K27triM may be a novel epigenetic mechanism for miR-22 silencing in ALL.

Disclosures

No relevant conflicts of interest to declare.
Fluorescent PCR Based Gene Dose Assessment for Detection of Deletion Mutations in the FIX Gene Among Carriers of Hemophilia B.
Authors: Edison Eunice Sindhuvi , Devi G. Sankari , Jayandharan G. , Velayudhan Shaji R , Viswabandya Auro , Mathews Vikram , Chandy Mammen , Srivastava Alok
Published: October 2019

Abstract 3486

Poster Board III-423

Haemophilia B (HB), an X linked inherited disorder is caused by heterogeneous mutations in the F9 gene. Approximately 3% of hemophilia B patients have major deletions in the F9 gene. Gross and small deletions in the F9 gene in HB affected males are easily detected by PCR but detecting the carrier state of females in the family is challenging due to the presence of the normal allele. Different methods like linkage analysis, real time PCR and MLPA have been used to assess the carrier status in this situation. Linkage analysis is limited by the availability of informative markers and adequate number of family members. Real time PCR involves standardisation and preparation of calibration curves for each run. Although MLPA is a better alternative, it can be time consuming and involves multiple steps. We have therefore developed a fluorescent PCR based gene dosage approach which is simple, rapid and cost-effective for determining the carrier status of females in families with deletions in the F9 gene. 200ng of DNA extracted by standard protocols was used for amplification with primers designed to amplify a 160bp product from exon h in the F9 gene. One of the primers was fluorescently labelled. Amplification was carried out using these primers for 20 cycles only and the amplified product was subjected to capillary electrophoresis on an ABI 310 genetic analyser. A 230 bp fragment in the albumin gene was used as the control. Analysis was done using Genescan and Genotyper software. The ratio between the peak heights of the exon h in the F9 and control genes in the patient samples were normalised to a normal control. Five families with deletional HB were analysed (in toto deletion-1; Ex g-h – 1; Ex g-poly A-1; Ex h-poly A-2). The ratios in the probands and the family members are presented in the table. Out of eight females analysed, 6 were carriers and 2 were normal. The mean ratio in the carriers was 0.49±0.08 and 0.75±0.05 in the normal. Deletions are not uncommon in HB and deletions involving the exon g and h constitute a major group. Among 212 families with HB assessed at our centre, we have identified large deletions in 8 families (3.7%). It is interesting to note that all except one of these deletional mutations involved exon h. This method confirmed the presence of these deletions in the males and helped us to identify the carrier status of the females in the family.

Identification of carrier status of females with deletions in F9 gene by gene dosage Subject ID Peak height of Exh in F9 Peak height of albumin Normalised Ratio Interpretation HB5 284 442 0.59 Carrier HB6 305 489 0.57 Carrier HB22 188 372 0.47 Carrier HB63 85 165 0.48 Carrier HB129 247 295 0.78 Normal HB238 94 326 0.4 Carrier HB280 372 679 0.77 Normal HB384 202 670 0.4 Carrier

Disclosures:

No relevant conflicts of interest to declare.
A Comparison of the Sex-Specific Incidence of Immune Thrombocytopenia in Children, Teenagers, and Adults in the General Practice Research Database.
Authors: Yong Mellissa , Schoonen W. Marieke , Li Lin , Kaye James A.
Published: October 2019

Abstract 3502

Poster Board III-439

Background

Immune thrombocytopenia (ITP) is an autoimmune condition characterized by low blood platelet counts. It has been reported that the epidemiology of ITP in the pediatric population (<18 years old) is different from that in adults. We therefore assessed the incidence of ITP in the pediatric population as compared to the adult population in a large UK-based primary care database – the General Practice Research Database (GPRD).

Methods

We identified all patients with a first-time diagnosis of ITP recorded in the GPRD during the period from 1990 to 2005. Age- and sex-specific incidences of ITP were estimated as the number of patients with a first-time recorded diagnosis of ITP divided by the person-time contributed by all subjects in the database who did not have a diagnosis of ITP.

Results

Of the total of 1,146 incident cases of ITP among 29.2 million person-years (PY) of observation from 1990 to 2005, we observed 257 incident cases of pediatric ITP (<18 years old) in 6.1 million PY, of whom 197 were children (aged less than 10 years) and 60 were teenagers (10 – 17 years old) at diagnosis. The age-specific incidence estimate for ITP in children was 5.7 per 100,000 PY [95% confidence interval (CI): 5.0 – 6.6 per 100,000 PY], with a statistically significantly 60% higher incidence in boys compared to girls. By contrast, among teenagers the overall incidence was lower [2.2 (95% CI: 1.7 – 2.9) per 100,000 PY] with a suggestion of a 50% higher incidence rate in girls compared to boys. The overall incidence in the adult population was 3.8 (95% CI: 3.6 – 4.1) per 100,000 PY, with women having a statistically significantly 48% higher incidence compared to men.

Conclusions

Our results showed an interaction between age and sex with ITP incidence, suggesting that patterns of disease burden differ among children, teenagers, and the adult population. Further investigation into these differences, including comorbidities and treatment practices is warranted.

Financial disclosure

This study was supported by Amgen Inc.

Disclosures:

Yong: Amgen Inc.: Employment, Equity Ownership. Schoonen:Amgen Inc.: Employment, Equity Ownership. Kaye:Amgen Inc.: Research Funding.
Bone Marrow Cell Rejection by Innate Immune Pathways Is Exclusively Dependent On the TRIF Pathway.
Authors: Xu Hong , Yan Jun , Zhu Ziqiang , Hussain Lala-Rukh , Huang Yiming , Wen Yujie , Ildstad Suzanne T
Published: October 2019

Abstract 3526

Poster Board III-463

Research on transplantation rejection, tolerance, and immunosuppressive agents has been directed toward the adaptive immune response as, until now, it has been believed to be the dominant mechanism for alloreactivity. The role of innate immunity in transplantation has not been fully defined. Despite the sentinel role of innate immunity in driving and shaping adaptive immunity, the contribution of innate immunity to bone narrow cells has only recently been identified. TLR4-driven MyD88-dependent immunity has been demonstrated to be critical for allogeneic innate immune responses to solid organ transplants. Whether innate immunity plays a role in BMC rejection has not been defined. In the present studies we tested whether the absence of MyD88 would enhance allogeneic engraftment in bone marrow transplantation (BMT). The core component of TLR signaling is activation of an IL-1-like pathway dependent upon the adapter MyD88. MyD88−/− B6 mice (n=6) were used to study the role of MyD88 signaling in allogeneic BMC rejection. As TRIF is another crucial adapter for TLR3 signaling and responsible for induction of signaling via type 1 IFNs, B6 TRIF−/− mice (n=6) were also investigated. Wild-type B6 (H2b, n=6) mice were used as controls. In the present study, MHC plus minor antigen-disparate BALB/c (H2d) mice served as allogeneic bone marrow donors. Recipients were nonmyeloablatively conditioned with anti-CD154 mAb (day0 and +3) to block CD40:CD154 co-stimulatory pathway and sirolimus (day0 to +4) to block late-stage T cell activation by inhibiting IL-2 responsiveness. Recipients were transplanted with 15 × 106 allogeneic (BALB/c; H2d) marrow cells 4-6 hours following conditioning with 100 cGy total body irradiation (TBI). Allogeneic engraftment was achieved in 33.3% of MyD88−/− recipients at 1 mo after BMT, a percentage similar to that for wild-type B6 mice. Surprisingly, 100% of TRIF−/− mice engrafted. The level of donor chimerism in TRIF−/− mice was 5.1 ± 0.6% at one month after BMT, which was significantly higher than in MyD88−/− (1.8 ± 1.0%; P < 0.0001) or in wild-type B6 mice (1.3 ± 0.8%; P < 0.005) and thereafter through all time points tested (2, 4 and 5 months, P < 0.05). The levels of donor chimerism increased with time and the level of donor chimerism reached 14.7 ± 7.1% TRIF−/− at five months. These results suggest that allogeneic BMC rejection is uniquely independent of the MyD88 pathway but rather involves TRIF signaling. These data suggest that the innate immune response to BMC differs from that for solid organs and may reflect the specificity of BMT compared with other transplants. These findings demonstrate for the first time that BMC elicit an innate immune response via the TRIF pathway and may open the door to novel approaches for immune-based conditioning to promote engraftment.

Disclosures:

Ildstad: Regenerex: Equity Ownership.
NK Cell-Mediated Rejection of Bone Marrow Cells- In Vivo Evidence of NK Cell Subset Licensing.
Authors: Sun Kai , Barao Isabel , Redelman Doug , Murphy William
Published: October 2019

Abstract 3537

Poster Board III-474

Natural killer (NK) cells have been shown to attack virally-infected and transformed cells as well allogeneic bone marrow cells (BMC) but not normal self-tissues. The mechanism of missing self recognition and self tolerance of NK cells is poorly understood. NK cells exist as subsets based on expression of inhibitory receptors (Ly49 in mouse, KIR in man) that bind MHC class molecules. In vitro data have shown that murine NK cell subsets bearing Ly49 receptors for self MHC class I molecules have intrinsically higher effector function, supporting the hypothesis that NK cells undergo a host MHC class I-dependent functional education, allowing the NK cells bearing the appropriate Ly49 receptors to preferentially mediate effector function. Thus far, no in vivo evidence for this preferential licensing or arming has been shown. We assessed the intrinsic response potential of the different Ly49+ NK cell subsets in BMC rejection without having the complicating effects of binding MHC on the target cell (which delivers potent inhibitory signals to the NK cell) by using β2-microglobulin deficient (β2m−/−) mice as donors which totally lack class I MHC molecules. Using syngeneic, congenic, and allogeneic strains of mice as recipients and depleting the different Ly49 subsets, we found that NK cell subsets whose Ly49 molecules have been shown to bind “self-MHC Class I” (ie Ly49G2 in H2d and Ly49C in H2b haplotypes) were found to be the dominant subset responsible for mediating the rejection of the β2m−/− BMC. This provides the first in vivo evidence for host MHC class I-dependent functional education (licensing or arming). Importantly, we also demonstrated that prior activation of the NK cells in vivo could override this licensing effect and allow the “non-licensed subset” to mediate rejection. The pattern of NK mediated rejection ability by Ly49 subsets was observed in B10D2 allogeneic H2d strain mice but not observed in BALB/c allogeneic H2d strain mice indicating that licensing ability was not solely dependent on host MHC expression and supporting the role of MHC class I–independent system for NK-cell self-tolerance. Interestingly, all H2d strain mice (B10.D2, BALB/c, and B6D2F1) were able to resist significantly greater amounts of B2m-/- BMC than H2b strain mice indicating that the rheostat hypothesis regarding Ly49 affinities for MHC and NK cell function impacts BMC rejection capability. These results demonstrate that both MHC and non-MHC genes on the host has multiple effects on NK cell subset-mediated BMC rejection and that licensing or arming of Ly49 NK cell subsets can be observed in vivo.

Disclosures:

No relevant conflicts of interest to declare.