bluuebunny/crossref_metadata_2025 huggingface dataset:
Sirt1 Expression Related with Sarcoma Growth after Stem Cell Transplantation Combined with Depletion of Myeloid-Derived Suppressor Cells
Authors: Li Ming , Shi Ming , Cui Yunze , Adachi Yasushi , Ikehara Susumu
Published: October 2019

Sirtuins (Sirt) play a number of roles in various aging-related diseases such as obesity, diabetes and cancer, and Sirt1 plays a double role in cancer. The combination of bone marrow transplantation (BMT) and thymus transplantation (TT) has been shown to prevent the growth of malignant tumors. Myeloid-derived suppressor cells (MDSCs) can differentiate from bone marrow stem cells under pathological conditions such as cancer, and prevent the potent action of T cells and NK cells, thereby promoting tumor growth. Thus, depleting MDSCs is important when treating cancer. We therefore investigated the relationship between Sirt1 expression and sarcoma growth in sarcoma-bearing mice treated with BMT+TT and the depletion of MDSCs using Gr-1 Ab. Our results showed that only granulocytic MDSCs (G-MDSCs) were depleted by the Gr-1 Ab in these mice. Moreover, the sarcomas decreased significantly in size in the Gr-1 antibody group when compared with the BMT group. This was due to the increase in the percentage of T cells and decrease in the percentage of G-MDSCs. Furthermore, Sirt1 expression decreased significantly in the sarcoma. These findings suggest that inactive Sirt1 may prevent sarcoma growth in sarcoma-bearing mice that receive stem cell transplants and depletion of MDSCs.

Disclosures

No relevant conflicts of interest to declare.
Eighteen Years Experience of Granulocyte Donations - Acceptable Donor Safety?
Authors: Axdorph Nygell Ulla , Sollén-Nilsson Agneta , Bjorkholm Magnus , Lundahl Joachim
Published: October 2019

Background. Granulocyte transfusions are given to patients with life-threatening infections, refractory to treatment. The donors are stimulated with corticosteroids +/- G-CSF. However, data regarding the donor’s safety is sparse. The objective was therefore to evaluate short and long term adverse events (AE) in stimulated donors.

Study design and methods. All consecutive granulocyte donors 1994-2012 were identified through our registry. Data were evaluated for the number of aphereses, stimulation therapy, AE, blood values post donation and recent status from the donation records.

Results. 154 volunteer donors were mobilized for 359 collections. Age at first granulocyte donation was 43 years (median; range 19-64 years). Follow-up was 60 months (median; range 0-229 months). The dose G-CSF per collection was 3.8 ug/kg body weight (median; range 1.6-6.0 ug/kg). Sedimentation agent was HES. Short term AE were mild. Blood values 4 weeks post donation with minor reductions/elevations mostly resolved in later donations. Fourteen donors were excluded from the registry due to hypertension (4), diabetes (2), atrial fladder (1), ca mammae (1), ca in situ urethra (1), MGUS (1), thrombosis (1), anaphylaxis (1), primary biliary cirrhosis (1), unknown (1). Three donors are deceased due to diabetes, acute myocardial infarction and unknown cause. Time from first donation to exclusion was 37 months (median; range 1-198). All donors except one were excluded/died at least 6 months after the first granulocyte donation.

Conclusion. No serious short term AE were observed. Due to the variability of diagnoses among excluded /deceased donors, we propose that it is less likely that granulocyte donations have a causative impact on these donors’ exclusion or death.

Disclosures

No relevant conflicts of interest to declare.
A Novel Protocol Decreased Incidence of Hepatic Veno-Occlusive Disease and Improved Results in Hematopoietic Cell Transplantation for Patients with b-Thalassemia Major
Authors: Li Chunfu , Pei Fuyu , Li Qi , Liao Jianyun , An Shengli , Wu Xuedong , He Yuelin , Feng Xiaoqin , Zhou Xiaohui , Shi Lei , Liu Huaying , Li Na , Xu Wenfeng , Tan Wanxia , Peng Zhiyong , Chen Jiaqi , Ren Yuqiong
Published: October 2019

Hepatic veno-occlusive disease (VOD) is a common complication of hematopoietic cell transplantation (HCT), especially patients with β-thalassemia major (TM). To estimate whether incidence of VOD recently decreased after using NF-08-TM protocol (NP), 311 TM-HCTs performed from February 2003 to June 2013 were analyzed. 241 patients received NP in or after 2009 and 70 received non-NP before 2009. VOD was diagnosed by Seattle criteria (SC) or Baltimore Criteria (BC). Patients were stratified by Nanfang (NF) criteria. A total of 31(10.0%) and 14 (4.5%) HCTs developed VOD (6 and 5 developed severe VOD) in SC and BC cohorts, respectively. The incidence of VOD was significantly lower in NP versus non-NP groups and in Class 2 versus Class 3 groups. Overall survival was significantly higher in NP versus non-NP cohorts. Rate of VOD in alternative donor transplant (ADT) was similar to that in matched sibling transplant (MST). Requirement of platelet and value of D-dimer significantly increased in the VOD patients. Our study showed the incidence of VOD significantly decreased in our center after using NP. ADT was similar to MST on rate of VOD. NF criteria of stratification can indicate occurrence of VOD. The SC can be more suitable criteria for early diagnosis.

Disclosures

No relevant conflicts of interest to declare.
Clinical Study of Cytomegalovirus Infection after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: He Jun , Ding Jiahua , Ni Ming , Chen Baoan
Published: October 2019

To analysis the cytomegalovirus（CMV） infection rate and median time which the CMV-DNA was positive for the first time. Simultaneously, to demonstrete the risk factors associated with CMV infection and CMV disease distribution and mortality after allogeneic hematopoietic stem cell transplantation（allo-HSCT).A retrospective study among 108 patients with allogeneic hematopoietic stem cell transplantation allogeneic hematopoietic stem cell transplantation(allo-HSCT) was conducted during 1st Jan 2004 and 31stMar 2014 in Zhongda hospital.The infection rate of CMV was 52.78%(57/108), the median time of CMV infection was 44d, especially during 30d~100d. The univariate analysis showed that CMV infection rate was associated with the HLA-identical situation between the donor and recipient, whether the use of Anti-human thymus globulin(ATG) conditioning regimen, neutropenic period after transplantation exceeded 10d and graft-versus-host disease (GVHD). Multivariate analysis showed that CMV infection rate was associated with neutropenic period after transplantation which exceeded 10d and graft-versus-host disease (GVHD).The mortality of the patients who had CMV infection was 10/17 and the mortality of CMV interstitial pneumonia was higher. CMV infection was one of the most commonly happened infection after allo-HSCT.

To reduce the incidence of CMV disease and mortality, the best choice of Allo-HSCT was HLA-identical donor, ATG should be used during the process, and neutropenic period should be reduced less than 10 days. Moreover, it is necessary to strengthen the treatment of CMV infection actively.

Disclosures

No relevant conflicts of interest to declare.
Low Percent of Granzyme B Positive T-Regulatory Cells (CD4+CD25high) As a Predictor of Acute Graft-Versus-Host Disease (GVHD) after Allogeneic Hematopoietic Stem Cell Transplantation (allo-HSCT)
Authors: Drokov Mikhail , Parovichnikova Elena N. , Kuzmina Larisa A , Galtseva Irina , Vasilieva Vera , Savchenko Valeri G.
Published: October 2019

Introduction. Granzyme B is a serine protease commonly found in the granules of cytotoxic lymphocytes and natural killer cells. It is secreted with the pore forming protein perforin and mediates apoptosis in target cells. Granzyme B – mediated cytolysis is one of the regulatory mechanisms (together with IL-2 receptors (CD25)) by which T-regulatory cells (Tregs) influence on T-effectors cells. It is a well-known fact that Tregs participate in a pathogenesis of aGVHD and their amount after allo-HSCT inversely correlates with the probability of aGVHD incidence. No clinical data exist regarding the Granzyme B positive Tregs in this setting.

Patients and methods. Peripheral blood samples were collected in EDTA-tubes at day +30, +60, +90 after allo-HSCT and at day of acute GVHD onset from 27 patients with hematological malignancies (AML=12, ALL n=12, LPD=2, MDS =1; 6 with active disease, 21 - in CR) after allo-HSCT (from matched unrelated donor n=23, from matched related donor n=4; MAC = 10, RIC=17).

The anti-CD4-APC-Cy7, anti-CD25-FITC and anti-Granzyme B-PE (Becton Dickinson, USA) antibodies were used to determine T-regulatory cells population as CD4+CD25high. We did not include anti-FoxP3 antibodies as FoxP3 is not so specific in humans and particularly after allo-HSCT due to technical difficulties, several isoforms, FoxP3-positivity on activated nonregulatory CD4+ cells. CD4- lymphocytes (CD8+ and NK-cells certainly containing granzyme B) were used as internal positive control to define the percent of CD4+CD25highGranzymeB+ cells and gMFI (geometric mean fluorescence intensity) among the mononuclear cell fraction. 50000 of CD4+ cells were analyzed on a BD FACSCanto II (Becton Dickinson, USA).

Results. 11 patients (42%) have developed aGVHD (II-IV) at a median time of +26 day (8 - 88) after HSCT. The percent of CD4+CD25highGranzymeB+ cells among CD4+cells at day +30 after allo-HSCT in patients who never developed aGVHD was statistically higher (9,49±2,79; p=0.003) than in patients at day of aGVHD onset (3,8±1,78). It’s worth to note that in 4 patients who did not have signs of a GVHD on day +30 but developed it later (on day +40, +45, +74, +88), the percent of CD4+CD25highGranzymeB+ cells at day +30 was as low (3,38±1,47) as at day of aGVHD onset and significantly lower (p=0.003) than in patients who never developed aGVHD, thus predicting the occurrence of aGVHD in the future.

Conclusion Despite the fact that the analyzed group is small, we suggest that low relative amount of CD4+CD25highGranzymeB+ cells after allo-HSCT may contribute to the development aGVHD and even predict it onset. This fact, of course, needs further investigation.

Disclosures

No relevant conflicts of interest to declare.
Single Center Retrospective Comparison of Gvhd Preventive Regimens after Cy/Flu Minimal Intensity Allotransplantation
Authors: Ranganath Praveen , Robertson Michael J. , Abonour Rafat , Schwartz Jennifer E. , Farag Sherif , Wood Lisa l , Kiel Patrick , Nelson Robert
Published: October 2019

Graft-versus-host disease (GVHD) remains the 2nd leading cause of mortality—after relapse/progression following allogeneic hematopoietic cell transplantation for the treatment of hematological malignancies. The incidence of clinically significant GVHD ranges from 10 to 80% and accounts for approximately 10-20% transplant related mortality. The combination calcineurin inhibitor and methotrexate is widely used as a GVHD preventive following myeloablative conditioning; however, there is no standard prophylaxis regimen for Cy/Flu minimal intensity nonablative transplants. The objective of the current study is to compare outcomes in patients treated at a single institution with identical conditioning regimens and supportive methods but different GVHD preventive regimens.

Outcomes on 134 patients who underwent nonmyeloablative transplant between May 2000 and Nov 2011 were retrospectively analyzed. The primary endpoint was incidence and severity of acute GVHD (aGVHD). Secondary endpoints included incidence and severity of chronic GVHD (cGVHD), incidence of relapse, overall survival (OS), progression-free survival (PFS) and causes of death.

Median follow-up was 49 months; the median age at transplant was 56 years, 66% of patient were not in remission at time of transplant (predominately myelodysplasia). Donors were peripheral blood mononuclear cell (PBMC, n=133) or bone marrow (n=1) HLA matched related or partially matched (9/10) donors. GVHD preventive regimens included the following: (1) single agent cyclosporine (CsA); (2) combination CsA/mycophenolate mofetil (MMF); (3) CsA+ basiliximab after ANC > 500/mm3; (4) CsA + basiliximab post-transplant, pre-engraftment; (5) sirolimus + tacrolimus.

The incidence of grade III-IV acute GVHD was lower in patients who received sirolimus/tacrolimus than patients in the cyclosporine group (25% and 32%, respectively; p = .062). There was a significantly higher proportion of patients in the cyclosporine group who had clinical extensive chronic GVHD (62% vs. 25%, p = .008). The incidence of relapse trended higher in the sirolimus/tacrolimus group compared to CsA alone or CsA/MMF (50% vs. 28% vs. 30%, respectively, p= .519). There was a trend towards slightly better OS and PFS with the cyclosporine-based prophylaxis, but follow-up was longer with these regimens.

Sirolimus/tacrolimus provides potent GVHD prophylaxis for nonmyeloablative Cy/Flu allogeneic transplantation; however, may be associated with higher relapse/progression rates. Larger numbers and/or comprehensive analyses of multicenter registry data might help further delineate differences in clinical outcomes between these regimens.

Figure 1 Figure 1.

Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
Outcome of Allogeneic Hematopoietic Cell Transplantation in AML or MDS with a HLA-Mismatched Unrelated Donor: A Single Center Analysis
Authors: Schmohl Jörg , Dorn Christiane , Haen Sebastian P. , Faul Christoph , Kanz Lothar , Bethge Wolfgang
Published: October 2019

Introduction: In acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS), allogeneic hematopoietic cell transplantation (HCT) can constitute a curative treatment option. Best results are reported using 10/10 HLA-matched donors, and several reports indicated inferior outcomes because of increased graft versus host disease (GVHD) and non-relapse mortality (NRM) if a HLA-mismatched donor is used.

Methods: Here we present a single center analyses evaluating all patients (pts) with AML or MDS receiving allogeneic HCT with a mismatch unrelated donors (MMUD) between 2004 and 2014.

Results:137 pts were evaluated. Median age was 51 years (range 18-76). For myeloablative conditioning (n=52) the following regimens were used: 12 Gy total body irradiation (TBI) and high-dose cyclophosphamide (n=40), or high-dose busulfan and cyclophosphamide (n=8); for reduced intensity conditioning (n=83) fludarabine with either busulfan (n=27), melphalan (n=6), treosulfan (n=7), cyclophosphamide (=15) or TBI (n=19) was used. Stem cell sources were either peripheral blood (n=134) or bone marrow (n=3). The donor/recipient match was a 9/10 alleleic mismatch in 41% and a 9/10 antigen-mismatch in 59% of cases. All but 21 patients received antithymocyte globuline prior to transplantation for GVHD prophylaxis. Post-transplant immunosuppression consisted of a combination of calcineurine inhibitors with either methotrexate (n=65) or mycophenolate mofetil (n=28). Kaplan-Meier estimated overall survival (OS) for all patients at 1 year was 70% and at 3 years 23%. Cumulative incidence of relapse was 32%. Median follow up of patients alive was 40 months (4-122 month). Engraftment with >500 neutrophils and >25.000 platelets was observed at a median of 20 and 20 days after HCT, respectively. Cumulative incidence of NRM until day 100, 1 year and 3 years was 16%, 27% and 28%, respectively. OS and NRM were not different in pts with allelic or antigen-mismatched donors (OS at 1 year/3 years: 65%/48% versus 48%/33% (p=0,2411), NRM at day 100, 1 year and 3 years: 21%, 24% and 33% versus 20%, 27% and 37% (p=0,6210) ). Acute GvHD mostly occurred at lower grades (grade I 34%, grade II 16%, ≥ grade III 10%). Risk of acute GVHD for patients with an antigen-mismatched donor was not different compared to those with an allelic-mismatched donors (51% versus 42%, p=0,4086).

Conclusion: Stem cell treatment with mismatched donors in allogeneic HCT is a valuable and safe option for pts with missing matched stem cell source. Due to optimized pre- and-posttransplant treatment, satisfactory long term survival can be achieved.

Disclosures

No relevant conflicts of interest to declare.
Outcome in Allo- SCT in Patients with Refrectory Acute Lymphoblastic Leukemia with Non TBI Conditioning Regimens
Authors: Sharma Ajay , Nair Velu , Das Sataya Ranjan , Sharma Sanjeevan , Singh Jasjit , Kumar Rajeev , Kotwal Jyoti , Kaur Jasmit
Published: October 2019

Background: Despite intensive remission induction chemotherapy, up to 5–10% AYAs with acute lymphoblastic leukemia (ALL) are primarily refractory.1,2 Of those that achieve a remission, as much as 70–80% fail consolidation largely due to relapse or sustained aplasia. Allogeneic stem-cell transplantation (allo-SCT) remains the only salvage option with curative potentials available, but the long-term benefits of the procedure and whether all patients should benefit from it remains controversial. There has always been a discussion about best conditioning regimen for ALL transplants especially in resource limited situation where TBI is not available. We analyzed our data on allo-transplants for ALL, wherein we had used non-TBI protocols for conditioning.

Study: This study retrospectively analyzes the outcome in 32 patients ( 19 females & 13 males) who underwent Allo-SCT for refractory acute lymphoblastic leukemia at our center between 2000-2013. The median age was 28 years (range, 7–26). 14 patients had relapse, 8 failed re-induction, 5 in partial remission & 7 patients had high risk disease ( Ph+ disease). All patients were in a state of bone marrow remission at the time of transplantation. All received myeloablative conditioning using busulfan( BU) & cyclophosphamide( CY). The stem cell harvesting was done by peripheral G-mobilized stem cell collection from related donors. All patients received graft-versus-host disease (GVHD) prophylaxis with cyclosporin and methotrexate. Infection prophylaxis & supportive care was provided as per standard protocols. The median cell dose was 5.4 MNCx10 7/Kg (3.8-7.6) & CD34 cells 3.7x105/Kg. 31 patients engrafted successfully by median day 12(7-23) for White cells & median day 38 ( 27-48) for platelets. One patient experienced late graft failure. 21 patients developed Gd-II acute-GVHD, 11 of them progressed to severe form( Gd-III). 5 pateints had severe grade IV Ac GVHD. 3 of them died chronic-GVHD appeared in 14 patients,. 19 patients (58%%) remain alive after a median of 60 months (range, 6–174); with 13 deaths, mostly from relapse (n=10) and infections (n=3). Overall survival (OS) and progression-free survival (PFS) at 5 years was 53% and 48%, respectively. OS and PFS were significantly better with blasts 20% and time to transplant 1 year.

Conclusions: We conclude that patients with refractory leukemia can benefit from allogeneic BMT, if they are transplanted in a state of remission, by using non-TBI based protocols. Thus in a resource limited situation, the outcome in refractory ALL can still be improved by improvisation of protocols without using TBI in conditioning regimens.

Disclosures

No relevant conflicts of interest to declare.
Analysis of Racial Disparity in Plasma of Healthy Volunteers Using Rotational Thromboelastometry Reveals Higher Prothrombotic Profile in African Americans
Authors: Janbain Maissaa , Liszewski Walter J , Leissinger Cindy A
Published: October 2019

Racial differences in the incidence of arterial and venous thrombotic events are well established in literature, with a higher incidence noted in African Americans compared to Caucasians. Several studies have tried to explain this difference, by looking separately at chemical biomarkers, socioeconomic and clinical risk factors. Yet the exact reason behind this disparity remains unclear. At least one study of thrombin generation suggested that African ethnicity was associated with increased peak thrombin generation when compared to Caucasians. Rotational thromboelastometry is a visco-elastic methodology that offers a global assessment of hemostasis using either whole blood or plasma. We explored the racial differences in rotational thromboelastometry findings using plasma samples from healthy volunteers.

We studied a cohort of 9 otherwise healthy adult volunteers with no history of cardiovascular nor thromboembolic events, 5 African Americans and 4 Caucasians. After informed consent, we collected citrated whole blood samples and processed them within 3 hours of phlebotomy. Platelet free plasma, obtained after centrifugation of whole blood for 20 minutes, was kept frozen at -70°C, and then thawed at 37°C for 5 minutes prior to testing. Samples were re-calcified with star-tem® reagent, and then the in-tem® reagent was added. The latter contains an optimized concentration of ellagic acid and partial thromboplastin phospholipid from rabbit brain. Thromboelastometry parameters including Clot Formation Time, Alpha Angle, and Maximum Clot Firmness were determined. We then compared the data between the two study populations using parametric unpaired Student’s t-test.

Our results showed that the Clot Formation Time was higher in the plasma of Caucasians when compared to African Americans with a difference between means of 40.1 ± 4.4 seconds (p <.0001); the Alpha Angle was lower in Caucasians with a difference between means of 3.45 ± 0.3 degrees (p <.0001); and the Maximum Clot Firmness was lower in Caucasians with a difference between means of 9.75 ± 2 mm (p <.01). These findings demonstrated that the plasma of Caucasians took longer to reach the maximum firmness, and this maximum firmness was less than that reached in the plasma of African Americans. This reveals a significantly increased prothrombotic profile in the plasma of African Americans compared to Caucasians.

Despite the limited number of participants, the striking observed differences in the thromboelastometry parameters suggest that global assays may offer benefit in assessing thrombotic risks in disparate patient populations, as they incorporate multiple components of the hemostatic system. Higher numbers of subjects and assessment of both whole blood and plasma are indicated. We are currently in the process of expanding our cohort to confirm these preliminary results.

Disclosures

No relevant conflicts of interest to declare.
Health Risks Among Adults after a Prolonged Flaring Incident at the British Petroleum Plant in Texas City
Authors: D'Andrea Mark , Mitter Ajay , Reddy G. Kesava
Published: October 2019

Objective: Human exposure to benzene is associated with multiple adverse health effects including hematopoietic malignancies, respiratory irritation, and immune system alterations and central nervous system abnormalities. The purpose of this study is to examine health effects of a benzene exposure among adult subjects from a prolonged flaring incident at the British petroleum (BP) refinery in the Texas City, Texas.

Methods: The study included adults aged 18 years or older who had been exposed and unexposed to benzene. Using medical charts, clinical data including white blood cells (WBC), platelets, hemoglobin, hematocrit, blood urea nitrogen (BUN), creatinine, alkaline phosphatase (ALP), aspartate amino transferase (AST), and alanine amino transferase (ALT) in adults exposed to benzene were reviewed and analyzed and compared with unexposed adults.

Results: A total of 2213 adult subjects (benzene exposed, n=1826 and unexposed, n=387) were included. Benzene exposed subjects had significantly higher levels of WBC (X 103 per µL) counts (7.9±2.3 versus 6.8±1.6, P=0.0000) and platelet (X 103 per µL) counts (270.8±60.9 versus 242.5±53.7, P=0.0.0000) compared with the unexposed subjects. The mean serum creatinine levels (mg/dL) were also significantly increased in the benzene exposed group compared with the unexposed group (1.0 ±0.2 versus 0.8±0.2, P=0.000). Serum levels of ALP (IU/L) was significantly elevated in the benzene exposed subjects compared with the unexposed subjects (82.1±15.6 versus 71.8±8.2, P=0.000). Similarly, benzene exposed subjects had significantly higher levels of AST and ALT compared with those unexposed to benzene.

Conclusion: Together, the results of the study reveal that Benzene exposure from the prolonged BP flaring incident caused significant alterations in their hematological and liver markers indicating that adult subjects exposed to benzene may be at a higher risk of developing hepatic or blood related disorders.

Disclosures

No relevant conflicts of interest to declare.
LDL Discordance in Pediatric Survivors of Acute Lymphoblastic Leukemia
Authors: Rosen Galit , Shaibi Gabriel Q
Published: October 2019

Background: Survivors of childhood acute lymphoblastic leukemia (ALL) have a four-fold excess risk of mortality secondary to cardiovascular events compared to the general population. While it is well-known that LDL has a role in atherogenesis and subsequent cardiac events, recent studies consistently show that LDL particle number (LDLp) is a stronger, independent predictor of coronary heart disease in adults compared to LDL-cholesterol (LDLc), especially when the values are discordant. There is emerging evidence that LDL discordance also occurs in children. We studied this phenomenon in pediatric ALL survivors and identify associated risk factors.

Methods: Complete data were available from 64 patients enrolled in the Phoenix Children’s Hospital pediatric cancer survivorship registry. Patients were 12.9±3.5 years of age (range: 7.7-22.7 years) with a mean time off therapy of 5.5±2.6 years (range 2.3-11.4 years). LDLc and LDLp were assessed by NMR spectroscopy. Patients were assessed by a registered dietician for calcium, fat, and fruit intake as well as physical activity. Ideal LDLc and LDLp were defined as <160 mg/dL and <1600 nmol/L, respectively, based on American Heart Association guidelines for people with standard cardiac risk. Discordance was classified based on ideal vs. not-ideal status for each parameter.

Results: Mean BMI was 22.7±5.7 kg/m2 (range 14-38 kg/m2); mean BMI percentile was 73.1±26.1% (0.9-99.5%) with 42% of patients considered overweight or obese at the time of evaluation. Mean LDLc and LDLp were 79±25.5 mg/dL (29-164 nmol/L) and 1124±517 nmol/L (411-2837 nmol/L), respectively, with 89% (n=56) of survivors exhibiting ideal LDLc and 47% (n=30) of survivors exhibiting ideal LDLp. In a subgroup of patients with ideal LDLc, 40.6% (n=26) exhibited not-ideal LDLp and were classified as discordant. Regression analysis showed that after adjusting for age and gender, BMI (p<0.00001) and LDLc (p<0.00001) were significant independent predictors of LDLp while lifestyle factors and time off therapy were not significant predictors of LDLp. When patients were grouped as concordant (ideal LDLc and LDLp, n=30) or discordant (ideal LDLc and elevated LDLp, n=25), BMI was 17.6% higher in the discordant group (25.0±5.4 kg/m2 vs. 20.6±5.0 kg/m2, p=0.003).

Conclusion: In this population of pediatric ALL survivors, LDLc measurements may not provide a complete assessment of cardiovascular disease risk. Given that LDLp is an early predictor of cardiovascular outcomes, conventional lipid testing may underestimate the true cardiovascular disease risk in ALL survivors. Survivors may benefit from early, expanded screening and targeted intervention. Further studies should focus on the broader pathophysiology of cardiovascular disease in pediatric ALL survivors.

Disclosures

No relevant conflicts of interest to declare.
Periodic Monitoring of Components of the Metabolic Syndrome in Adolescent Survivors of Childhood Cancer
Authors: Lee Young-Ho , Shin HyeKyung , Kang Hye-Roung , Yang Seung
Published: October 2019

Purpose: The components of metabolic syndrome (MS) have been tended to increase and associated with cardiovascular risks in long-term survivors of childhood cancer. We investigated the prevalence of components of MS in adolescent survivors of childhood cancer and tried to monitor them periodically.

Materials and methods: We investigated 44 adolescent survivors of childhood cancer, median age 14.9 years (range 10–19.8 years) and median follow-up time elapsed after off-therapy 7.4 years (range 5–16.5 years). We measured body mass index (BMI), systolic and diastolic blood pressure, triglycerides (TG), high-density lipoprotein (HDL)–cholesterol, and fasting glucose. Fatty liver was evaluated by ultrasound examinations during follow-up period.

Results: No survivors demonstrated MS with 3 components, however, 18% of survivors (8/44) have 2 abnormal components and 43% (19/44) have 1 abnormal components, respectively. The frequency of each component was: increased BMI, 11%; elevated blood pressure, 0%; elevated TG level, 41%, low HDL cholesterol, 27%; and elevated fasting glucose, 0%. Among these components, the increased TG levels were highly prevalent in survivors than in general population (p=0.000). Fatty liver was identified in 8 survivors (18.2%). Sixteen of 44 survivors (36.4%) received at least 3 repeated examinations annually. Twelve out of 16 survivors have 1 or 2 abnormal components at initial examination. Their number of metabolic components have been shown to decrease in 4, persisted in 3, and increased in 5 survivors. Three of 4 survivors who have no components of MS at initial examination showed at least 1 abnormal component during follow-up.

Conclusions: We observed the high incidence of increased TG level in adolescent survivors of childhood cancer, and detected abnormal components of MS during periodic follow-up. Lifestyle interventions and periodic long-term follow-up monitoring would be needed to reduce the metabolic risks in childhood cancer survivors.

Disclosures

No relevant conflicts of interest to declare.
Bilateral Involvement Is an Independent Risk Factor in Testicular Relapse of Childhood ALL: Results of the Trials ALL-REZ BFM 83 - 2002
Authors: Chen-Santel Christiane , Tabien Ulrike , van Schewick Claudia , Schrappe Martin , Escherich Gabriele , Eggert Angelika , Henze Günter , Stackelberg Arend , Hundsdörfer Patrick
Published: October 2019

Introduction:

In childhood acute lymphoblastic leukemia (ALL) about 15-20% of patients suffer from relapse after frontline therapy. The testes are the second most frequent site of extramedullary relapse. Up to now there are no data on the impact of the extent of testicular relapse on prognosis.

Patients and Methods:

We retrospectively analyzed outcome of children with testis relapse of ALL who were treated according to ALL-REZ BFM trials from March 1983 until December 2013. Only patients with first relapse were included in the analysis. From a total of 1603 boys registered within the ALL-REZ trials since 1983, testicular involvement was identified in 302 patients. Local therapy consisted of orchiectomy (or irradiation at 24 Gray) of clinically involved testes. Clinically not involved testes were irradiated at 15 or 18 Gray in case of histologically proven leukemic infiltration. Most patients received intensive multi-drug chemotherapy followed by conventional maintenance therapy.

Results:

Extent and time point of relapse did not significantly differ between isolated testicular (n=135) and combined BM and testicular relapse (n=167). Isolated testicular relapse was associated with improved 5 year-event free survival (pEFS) compared with combined relapse (0.70±0.04 vs. 0.54±0.04, p=0.005). Bilateral testicular involvement accounted for a worse pEFS than unilateral involvement (0.48±0.05 vs. 0.71±0.04, p<0.0001). This was particularly evident in patients with isolated testicular relapse (bilateral (0.53±0.08) vs. unilateral relapse (0.79±0.05), p<0.0001), but not in combined relapse (bilateral (0.45±0.06) vs. unilateral relapse (0.62±0.06), p=0.11). Only in patients with very early/early isolated testicular relapse bilateral testicular involvement was associated with poor outcome (0.40±0.09 vs. 0.68±0.08, p=0.015), whereas no significant differences were detected in the late relapse group (0.79±0.11 vs 0.86±0.05, p=0.11). Multivariate analysis identified time point of relapse, extent of testicular relapse and combined relapse as independent risk factors in testicular ALL relapse. Subsequent local relapse was rare, ninty-eight patients (32.5%) suffered a subsequent, mostly BM relapse.

Conclusion:

Isolated testicular relapse is associated with improved outcome compared to combined BM/testicular relapse. Bilateral testicular involvement is an independent risk factor for relapse, particularly prognosis of patients with (very) early isolated bilateral testicular relapse was poor. As local in contrast to systemic subsequent relapse was rare, this group of patients might benefit from intensification of chemotherapy including allogeneic stem cell transplantation.

Disclosures

No relevant conflicts of interest to declare.
Ex vivo Fucosylation Enhances Regulatory T Cell Anti-Graft Versus Host Disease Potency in Mice
Authors: Parmar Simrit , Liu Xiaoying , Eric Yvon , zweidler-mcCay Patrick , Shah Nina , Rezvani Katayoun , McNiece Ian K. , Shpall Elizabeth J.
Published: October 2019

Emerging preclinical data indicate a potential therapeutic use of regulatory T cells (Tregs) to suppress or eliminate graft versus host disease (GVHD). Adoptive therapy with Tregs has been examined as a prophylactic strategy for GVHD. Thus far, 1:1 ratio of Tregs to conventional T cells (Tcons) has been required to prevent GVHD, and such high targeted cell doses are hard to achieve in a clinical setting due to variability in Treg frequency and expansion potential. As a result, novel strategies are needed to generate clinically-effective Treg products. Additionally, since the ability of Tregs to enter inflamed tissues has been shown to be critically dependent on their ability to bind E- and P-selectins, we sought to exploit this pathway to improve Treg homing. Incubation of culture expanded Cord Blood (CB) Tregswith fucosyltransferase-VI (FTVI) increased the degree of fucosylation from 8% to 64%. Importantly, fucosylated Tregs are able to suppress T cell proliferation in an in vitro allogeneic mixed lymphocyte assay and show preferential increase in the E-selectin binding ability when compared to untreated Tregs. We injected 10e6 Tregs vs. 10e6 fucosylated Tregs followed by 10e7 peripheral blood mononuclear cells in the sublethally irradiated NOD/SCID gamma null (NSG) xenogenic GVHD mouse model, and recipients of untreated Tregs showed weight loss as early as day 6 whereas fucosylated Treg recipients retained weight until day 12 (Fig 1). Fucosylated Tregs were detected in the peripheral blood of mice until day 31 as opposed to day 12 for untreated Tregs. As compared to recipients of untreated Tregs, lower numbers of GVH-inducing allogeneic T-cells were detected in the secondary lymphoid organs in the fucosylated Treg recipients (Table 1). The overall survival of fucosylated Treg recipients was significantly superior to that of untreated Treg recipients (Fig 2). We conclude that prophylactic treatment with fucosylated Tregs prevents GVHD and improves survival in a xenogenic mouse model at doses that are i) at less than 1:1 ratio with conventional T-cells (Tcon) and ii) are lower as compared to untreated Tregs. We believe that by overcoming the 1:1 dose requirement of Tregs: Tcons, we will be able to effectively translate Treg based adoptive therapy into the clinic for the prevention of GVHD. Establishing fucosylated Tregs as a preventive strategy for GVHD will result in a major breakthrough in the field of stem cell transplantation eliminating a major life threatening outcome of the therapy.

Figure 1 Figure 1.

Figure 2 Figure 2.

Figure 3 Figure 3.

Disclosures

No relevant conflicts of interest to declare.
PIM2 Pro-Survival Functions Are Mediated By RSK2 in AML
Authors: Hospital Marie-Anne , Jacquel Arnaud , Green Alexa , Lambert Mireille , Auberger Patrick , Lacombe Catherine , Hermine Olivier , Moura Ivan C , Mayeux Patrick , Bouscary Didier , Tamburini Jerome
Published: October 2019

Acute myeloid leukemia (AML) with FLT3 internal tandem duplication (FLT3-ITD) is a poor prognosis hematologic malignancy accounting for 30% of AML cases. Constitutive FLT3-ITD activation drives STAT5 signaling resulting in enhanced PIM kinases expression. PIM serine/threonine kinases (including PIM1,-2,-3) are involved in cell cycle and apoptosis regulation and thus represent emerging therapeutic targets. We previously reported an increased PIM2 protein expression in primary AML cells compared to normal CD34+ immature hematopoietic cells. Here, we aimed to study PIM kinases as potential therapeutic target in FLT3-ITD AML.

In two distinct FLT3-ITD+ human AML cell lines (MV4-11 and MOLM-14) doxycycline (Dox)-induced shRNA-mediated PIM2 knockdown enhanced apoptosis, attested by an increase in early apoptotic (annexin V positive, DAPI negative) and late apoptotic (both annexin V, DAPI positive) cells. Cell death upon PIM2 knockdown was confirmed by an inhibition of colony formation in methylcellulose. Mechanisms of apoptosis induction involved release of second mitochondria-derived activator of caspases (SMAC) as well as increased p53 and Bax expression and Bax nuclear translocation, leading to loss of mitochondrial membrane potential.

To gain further mechanistic insights, we performed global gene expression profiling in the MOLM-14 cell line lentivirally transduced with Dox-inducible PIM2 shRNA. Consistent with our functional analysis, cell-cycle regulatory genes (including c-MYC, CHK1 or PLK1) and pro-survival genes (including RSK2 or BCL2) were down-regulated, while pro-apoptotic genes (most notably TP53 and BAX) were up-regulated. Here we focused on RSK2, a member of the mitogen-activated protein kinase (MAPK) signaling pathway that has not been previously reported to be a target of PIM2. In the MOLM-14 cell line, PIM2 knockdown reduced RSK2 mRNA and protein levels. RSK2 knockdown using a Dox-inducible RSK2 shRNA induced apoptosis similarly to that observed following PIM2 knockdown in this cell line. In parallel with PIM2 knockdown, RSK2 down-regulation enhanced p53 expression and activity, as measured by increased expression of its transcriptional target p21, as well as Bax expression.

In summary, our results suggest that RSK2 is a PIM2 transcriptional target that contributes to PIM-2-dependent cell survival mediated by a novel RSK2-p53-Bax signaling pathway. RSK2 therefore warrants further study as a potential pharmacologic target in AML (particularly in cases resistant to PIM inhibition) as well as other PIM-addicted malignancies.

Disclosures

No relevant conflicts of interest to declare.
APP Gene Is Correlated with C-KIT Mutations and Indicates Poor Disease Outcome in AML1-ETO-Positive Acute Myeloid Leukemia
Authors: Yu Guopan , Jiang Ling , Meng Fan Yi , Yin Changxin , Wang Zhixiang , Zhou Hongsheng , Jiang Xuejie , Zhong Qingxiu , Cao Rui , Liao Libin
Published: October 2019

Amyloid precursor protein (APP) has been reported to be highly expressed in AML1-ETO-positive acute myeloid leukemia (AML1-ETO-positive AML), and we found it correlate with extramedullary infiltration regulated of by APP/ERK/MMP-2 signal pathway in our previous study. It is also known that C-KIT mutations highly expressed in AML1-ETO-positive AML and cooperates with full-length AML1-ETO to induce AML in mice. In this study we further described a close correlation of APP gene with C-KIT mutations, as well as APP related clinical and prognostic significance in 65 patients with AML1-ETO-positive AML.

65 cases of AML1-ETO-positive AML patients with median age of 30 years old, who were admitted to our hospital from February, 2006 to June, 2013 and made the diagnosis according to WHO2008 diagnosis standard, were enrolled into this study. APP expression in bone marrow cells before the first chemotherapy was assessed by quantitative reverse transcriptase (QRT)-PCR method. These cases were accordingly divided into APP-H group (n=33, with high level of APP by QRT-PCR) and APP-L group (n=32, with lower level of APP by QRT-PCR) according to median APP expression. Incidence of C-KIT mutations, clinical characteristics and prognosis including complete response (CR), overall survival (OS), and recurrence free survival (RFS) with median 35 (6-96) months followed-up was differentiated between the two groups. Furthermore, expression of APP and AML1/ETO fusion gene were simultaneously monitored at the time of 3, 6, 12 and 24 months or relapse after CR by QRT-PCR method.

The incidence of C-KIT mutations was significantly increased in the APP-H group, as compared with the APP-L group (39.4% versus 12.5%) and it was positively correlative with APP expression (rp=0.435, P=0.004). Of the 17 patients harboring C-KIT mutations, 13 patients overexpressed APP gene (P=0.014) (Figure 1). Clinically, APP-H patients exhibited significantly elevated white blood cells count, increased extramedullary infiltration (P=0.039 and P=0.019, respectively). Moreover, APP overexpression was related to low rate of two-cycle CR, RFS and OS (P=0.020, P=0.001 and P=0.029, respectively) (Table 1). In addition, the change of APP expression was consistent with that of AML1-ETO fusion gene monitored by QRT-PCR method at different status of leukemia, though APP expressed differently in different patients with the same AML1-ETO expression.

Taken together, these data suggest that APP gene is correlated with C-KIT mutations and indicates poor disease outcome and dynamic monitoring APP expression could be another choice of minimal residual disease monitoring in AML1-ETO-positive AML.

Figure 1 Figure 1.

Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
Genome Editing Toolbox
Authors: Scharenberg Andrew M.
Published: October 2019

Nucleases capable of making targeted breaks in genomic DNA are a core technology required for genome engineering, an emerging field of technology for making precise alterations in cellular genomes. Over the past ten years, four major platforms have emerged for generation of nucleases able to make targeted DNA breaks with a high degree of efficiency and specificity: homing endonucleases, zinc finger nucleases, transcription activator-like (TAL) effector nucleases, and RNA-guided nucleases. This talk will cover the biochemistry and platform-specific attributes of each type of nuclease, along with evolution/improvements in nucleases and related technologies and aspects of the practical implementation of nuclease technology for gene knockout and gene repair in primary hematopoietic cells.

Disclosures

Scharenberg: Pregenen Inc.: Equity Ownership, Membership on an entity's Board of Directors or advisory committees; Cellectis therapeutics: Consultancy.
Taking it to the Clinic: Genome Editing for Blood Disorders
Authors: Urnov Fyodor D.
Published: October 2019

The ability to engineer precise genetic modification of human stem cells offers the promise to extend the range of their potential therapeutic application. This possibility is now being realized via the use of zinc finger nucleases (ZFNs). ZFNs are customizable, sequence-specific endonucleases that can be designed to introduce a discrete cleavage event at any user-chosen location within the stem cell genome. This presentation will describe our progress in translating ZFN-modified hematopoietic stem/progenitor cells (HSPCs) from concept to clinic, focusing on preclinical proof-of-concept studies supporting the development of autologous, genome-edited CD34+ HSPCs as potential treatments for both HIV/AIDS as well as certain monogenic diseases of the blood, such as β-thalassemia.

Disclosures

Urnov: Sangamo BioSciences: Employment.
Hepcidin at the Nexus of Infection and Anemia
Authors: Drakesmith Hal
Published: October 2019

Humans have two main reasons for needing to control the amount and distribution of iron in the body. First, iron is used in fundamental physiological processes, including carrying oxygen, generating energy from oxygen, and in utilizing the energy for activities such as macromolecular synthesis and DNA repair. However, iron requirements are not stable over time; blood loss, growth, pregnancy, changes in diet and even altitude all have powerful and sometimes acute effects on our need for iron and how we route it to tissues that need it – for example, in the case of blood loss, iron is released for erythropoiesis to help replace lost red blood cells. The second reason to regulate iron is in response to infection; evidence for a critical influence of iron on the outcome of infections is broad. Sequencing of microbial genomes reveals significant investment into iron acquisition, allowing pathogenic organisms to obtain iron from multiple host sources. Iron is required for pathogen proliferation and increased availability of host iron, either through experimental administration in animal models or, in humans, due to genetic causes or because of nutritive or therapeutic iron supplementation, exacerbates infection or increases its incidence. Part of the innate immune response to infection is to deny iron to the pathogen, slowing its growth and giving more time for other arms of immunity to mobilize. The hypoferremia of infection helps to inhibit growth of microbes in the bloodstream, and may be a critical response to prevent potentially fatal septicemia. Hepcidin is the liver-encoded peptide hormone that allows the body to maintain iron homeostasis, to rapidly release iron for erythropoiesis, and to lock iron away from microorganisms in response to infection. The ability to integrate these activities lies in the unique sensitivity of hepcidin expression to diverse physiological inputs. Transcription of hepcidin is enhanced by signals deriving from iron accumulation (through BMP/SMAD signaling) and from immune mediators (IL-6, IL-22, IFN-a), and is suppressed by expanded erythropoiesis via erythroferrone. In animal models the relative strengths of these signals determine hepcidin synthesis, and thus determine iron absorption, release, and storage. Our lab has been focusing on understanding hepcidin regulation in human populations where infectious disease, anemia, iron deficiency and inherited red cell disorders are all prevalent, for example sub-Saharan Africa and the Indian subcontinent. This lecture will describe how hepcidin analysis has helped us to understand the complex etiology of childhood anemia in malarious regions, suggested methods to improve iron therapeutic strategies, and indicated ways to use hepcidin diagnostically. The latter applications are now beginning to be tested clinically at the University of Oxford.

Disclosures

No relevant conflicts of interest to declare.
Regulation of Hematopoiesis by Cholesterol
Authors: Tall Alan R.
Published: October 2019

Leukocytosis is a risk factor for athero-thrombotic disease in humans, and develops in animal models of atherosclerosis in response to feeding high-fat, high-cholesterol diets. The ATP binding cassette transporters ABCA1 and ABCG1 promote cholesterol efflux to apoA-1 and high density lipoprotein (HDL), respectively and are targets of liver X receptor (LXR) transcription factors. Mice lacking ABCA1/G1 develop a dramatic myeloproliferative phenotype with monocytosis and neutrophilia, associated with expansion and proliferation of hematopoietic stem and myeloid progenitor populations (HSPCs). The transporters are highly expressed in HSPCs where they act to control proliferative responses to growth factors (IL-3, GM-CSF) by regulating plasma membrane lipid rafts and cell surface expression of the common β subunit of the IL-3/GM-CSF receptor. ABCG4 is closely related to ABCG1 but is expressed primarily in the megakaryocyte progenitor (MkP) population of the bone marrow. ABCG4-deficient mice have MkP proliferation and expansion, thrombocytosis, increased platelet/leukocyte aggregates and accelerated atherosclerosis. ABCG4 promotes cholesterol efflux onto HDL, and thereby reduces the cell surface expression of the thrombopoietin (TPO) receptor. This appears to involve membrane cholesterol enrichment and interruption of a negative feedback loop involving the TPO receptor and mediated by Lyn Kinase and c-CBL which mediate ubiquitination and internalization/degradation of the receptor. Overall results suggest that ATP binding cassette transporters promote cholesterol efflux, decrease membrane lipid raft formation and enhance the feedback downregulation of growth factor receptors in response to growth factor binding, with anti-proliferative responses that may be beneficial in atherosclerosis and myeloproliferative neoplasms.

Disclosures

No relevant conflicts of interest to declare.