bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulatory T Cells Preserve Anti-Viral Immunity While Preventing Lethal Graft-Versus-Host Disease
Authors: Winkler Thomas H. , Seefried Martina , Kroeger Irena , Hoffmann Petra , Edinger Matthias , Ullrich Evelyn , Mach Michael
Published: October 2019

Graft-versus-host disease (GvHD) is a frequent and life-threatening complication of allogeneic hematopoietic stem cell transplantation (HSCT) and is therefore one of the main factors that limits the broad application of HSCT. Over the last decades, several studies have reported a clinical association between GvHD and reactivation of cytomegalovirus (CMV). Using a lethal murine GvHD model with major MHC mismatch (C57BL/6 -> Balb/c), we were able to demonstrate that recipients latently infected with murine CMV (MCMV) before transplantation showed recurrence of CMV infection concomitant with the manifestation of GvHD. Moreover, these preinfected recipients showed an accelerated mortality compared to recipients that were not preinfected. The therapeutic co-infusion of CD4+CD25+ regulatory T cells (Tregs) with conventional T cells (Tcons) prevented lethal GvHD in preinfected mice and, markedly reduced the recurrence of MCMV infection. Remarkably, these mice showed clearance of MCMV 5 weeks post transplantation in contrast to mice receiving only Tcons in which massive virus infection persisted. Enhanced reconstitution of T lymphocytes and establishment of an anti-MCMV antibody titer from donor B cells in these animals suggest that CD4+CD25+ Tregs do not interfere with an anti-viral response while suppressing Tcon-mediated GvHD. Therefore, our study revealed that the suppressive function of CD4+CD25+ Tregs is not affected by CMV reactivation and more importantly, that Tregs do not adversely affect the anti-viral immunity in the recipient. In sum, these results provide important information on the correlation of GvHD and CMV reactivation and underline the possible clinical benefit of Treg application in GvHD patients.

Disclosures:

No relevant conflicts of interest to declare.
Mobilization Scheme With Cytarabine In Poor Mobilizers Patients Diagnosed With Lymphoma
Authors: Calderón-Cabrera Cristina , González Magdalena Carmona , Martín Jesús , Herranz Eduardo Ríos , Novella Pilar Noguerol , De la Cruz Fátima , Carrillo Estrella , Espigado Ildefonso , Pérez-Simón Jose Antonio
Published: October 2019

Introduction

Mobilization regimens for hematopoietic stem cell transplantation have not undergone major changes except for the introduction of new drugs such as plerixafor. However, in patients receiving multiple lines of treatment, to obtain a suitable amount of hematopoietic progenitors (PH) is often difficult even with the addition of new drugs. We present our experience with a protocol of mobilization for poor mobilizer patients based on the use of cytarabine.

Material and Methods

From a total of 111 patients diagnosed with lymphoma who underwent autologous hematopoietic progenitors in our center from January 2005 to March 2013, 32 patients (35% of total) with previous mobilization failure, received cytarabine 400 mg/m2 /day x3 days + G-CSF (filgrastim or lenograstim) 10-16 mg /kg/day from day 7 to the end of collection. Cell blood cound and CD34 assays in peripheral blood were performed from day +11 postchemotherapy, and apheresis was started when CD34 > 10 x10e6/L.

Results

30 patients had non-Hodgkin lymphoma (NHL) (13 diffuse large B-cell lymphoma, 7 mantle cell lymphoma, 5 follicular lymphoma, 2 MALT lymphoma, 1 angioimmunoblastic lymphoma, 2 cutaneous lymphomas) and 2 Hodgkin's lymphoma (HL). Median age was 49 years (range 27-67). Median number of treatment lines prior to mobilization was 3 (1-5). In all 30 patients with NHL, an adequate amount of CD34 + cells was obtained. Only one patient with HL, who had previously received 5 lines of chemotherapy, did not mobilize. The average time from the end of chemotherapy to start of apheresis was 13.7 days (12-20). The average number of CD34 + cells circulating the starting day of apheresis was 41/μL (11-109). The mean of apheresis performed was 1.8 (1-3) with an average amount of CD34 obtained of 4.69 x10e6/kg (1.5-6.8). All patients received outpatient treatment and only 2 required hospitalization: one for short-term neutropenic fever and another due to a limited cutaneous hemorrhage during the neutropenia phase. All patients were transplanted without incidence and successfully engrafted. None required G-CSF early postransplant.

Conclusions

Mobilization with intermediate-dose cytarabine has proven to be a cost-effective regimen in lymphoma patients with prior mobilization failure.

Disclosures:

No relevant conflicts of interest to declare.
A Retrospective Analysis Of The Association Between Common Myeloablative Conditioning Regimens and Hepatotoxicity
Authors: Patel Rushang D. , Klumpp Thomas R. , Martin Mary Ellen , Kropf Patricia , Fasan Omotayo O. , Mangan Kenneth F.
Published: October 2019

High dose chemotherapy preceding stem cell transplant carries considerable risk of organ toxicity. Hepatic toxicity is one of the major causes of morbidity post transplant. The etiology is multi-factorial, with the common culprits being graft versus host disease as well as the agents used as part of the conditioning regimen. Evidence of pre-existing mild hepatic insufficiency demands an even more cautious approach while selecting a conditioning regimen prior to stem cell transplant. We analyzed the effect of various myeloablative conditioning regimens on post transplant liver function tests (LFT). CEP (cyclophosphamide, etoposide and cisplatin), BuCy (busulfan and cyclophosphamide), BEAM (carmustine, etoposide, cytarabine and melphalan) and Cy/TBI (cyclophosphamide with total body irradiation) were the four most commonly used regimens for autologous stem cell transplant (auto-SCT) at our center. A total of 404 patients were treated with CEP (210), BuCy (76), CyTBI (71) or BEAM (47) from 1988 to 2013. The majority of the patients underwent an auto-SCT for lymphoma (218 non-Hodgkin lymphoma, 98 Hodgkin’s lymphoma), while 67 transplant patients had leukemia. Patient age ranged from 17 to 73 years at the time of transplant, and the sex distribution revealed a male:female ratio of 1.5:1. We looked at the incidence of total bilirubin greater than 2 mg/dl, alkaline phosphatase greater than upper limit of normal, ALT greater than 126 U/L and AST greater than 82 U/L. The outcome of univariate analysis using chi-square test demonstrated statistically significant inter-group variability for three of the LFTs. CyTBI had the lowest incidence of hepatic insufficiency. Further analysis comparing CyTBI to the other three regimens, confirmed a highly significant statistical difference (p<0.01 by two-sided Fisher’s Exact test, see table below) in elevation of bilirubin, alkaline phosphatase as well as both the transaminases, favoring Cy/TBI as the safer regimen. This is an interesting finding given the common perception that TBI-based regimens are more toxic.CEP, BuCy and BEAMCy/TBIaffected/total (%)affected/total (%)p-valueTotal Bilirubin (≥ 2 mg/dl)27/263 (10.2 %)0/59 (0 %)0.007Alkaline Phosphatase (≥ upper limit of normal)97/262 (37 %)8/58 (13.7 %)0.0005ALT (≥ 126 U/L)39/263 (14.8 %)0/59 (0 %)0.0002AST (≥ 82 U/L)44/264 (16.7 %)2/59 (3.3 %)0.0065

Disclosures:

No relevant conflicts of interest to declare.
Impact Of Dysplasia On Outcome Of Patients With Acute Myeloid Leukemia and Response To Allogeneic Hematopoetic Stem Cell Transplantation
Authors: Farhan Shatha , Carroll Catherine , Pelland Danielle , Velasco Jose Carlos , Wautelet Susan , Ruemenapp Kenneth , Janakiraman Suseela M , Kuriakose Philip , Peres Edward , Janakiraman Nalini
Published: October 2019

Background

Myelodysplastic syndromes (MDS) are a heterogeneous group of malignancies characterized by dysplasia, cytopenia, ineffective hematopoiesis and by an increased risk of transformation to acute myeloid leukemia (AML). Recurrent aberrant karyotypes cannot entirely account for the genetic defects that are at the basis of the pathogenesis of MDS, as they are detected only in approximately 50% of patients.

Allogeneic hematopoietic cell transplantation (HCT) likely prolongs survival in patients with AML and MDS. In this report, we evaluated the impact of presence of dyspoesis in paients with AML with or without cytogenetic (CG) abnormality on outcome in our center.

Methods

We retrospectively reviewed all patients who were diagnosed with AML (non promyelocytic) in our center between 2002 and 2012. Primary objective was to study the impact of MDS with or without CG abnormalities on outcome of patients with AML. Demographics and disease-related variables were collected. OS was defined as the time from diagnosis to the time of death or last contact.

Results

Between 2002 and 2012, 123 patients with high or intermediate risk AML patients were treated at our center. Median age at diagnosis was 60 (range 19-89). Median OS for all patients was 368 days. Of 123 patients, 51 had MDS while 73 did not. CG abnormalities were present in 35 (68%) of patients with dyspoetic changes. Median age of AML patients with MDS was 59 while median age of AML without MDS patients was 55. Median number of blasts in bone marrow and peripheral blood in AML patients with MDS was 38% and 6% respectively. While median number of blasts in bone marrow and peripheral blood in AML patients without MDS was 67% and 39% respectively. Of 51 AML patients with MDS, 14 received HCT with median age of 56. Half of these received myeloablative regimen while the other half received reduced toxicity regimen.

The median survival time for AML patients without MDS was 401 days while the median survival time for AML patients with MDS was 278 days (p = 0.0201), Fig1.

For AML with MDS who received HCT, the median survival time was 586 days while it was 164.5 days for AML patient with MDS who did not receive HCT (p = 0.0013), Fig2.

Conclusion

In this small cohort from a single center, the results suggest that AML patients with dyspoetic changes do have a worse prognosis despite having lower percentage of blasts in bone marrow or peripheral blood. This can be explained by what Walter et al reprted, using next-generation sequencing, that the proportion of neoplastic marrow cells is indistinguishable in MDS and secondary-AML even with myeloblast count of zero. HCT can be performed in those patients including older patients with promising results.

Disclosures:

No relevant conflicts of interest to declare.
Autopsy Analysis On Epidemiology and Site of Involvement Of Invasive Fungal Infections (IFI) In Hematological Malignancies : A Retrospective Study at Hematologic Tertiary Care Department
Authors: Chierichini Anna , Monardo Francesca , Anaclerico Barbara , Borza Paola Anticoli , Bongarzoni Velia , Campagna Domenico , Cedrone Michele , Fenu Susanna , Norata Marianna , Paoloni Francesca , Ronci Benedetto , Annino Luciana
Published: October 2019

Background

Clinical diagnosis of IFI is difficult ,due to lack of sensitive and specific diagnostic tools. An assessment of trends concerning the prevalence of IFI is a challenge and postmortem data may be useful to monitor the local epidemiology ,the frequency and the disease patterns.

Aim

The aim of this retrospective analysis is to determinate the local epidemiology and the prevalence at autopsy of IFI, occurring in hematological malignancies at a single center  over a eleven years period.

Methods

We have retrospectively reviewed 161 patients – median age  62,5 yrs, range 22 -83 - with hematological malignancies, who underwent autopsy between 2002 -2012. Acute Myeloid Leukemia (AML) were 77, Acute Lymphoid  Leukemia (ALL) 11,  Lymphoproliferative disorders (LPD) 56 and other disorders 17. Acute leukemia pts received systemic antifungal  prophilaxis, whereas the others not absorbable prophilaxis. None patients received transplant procedures. An experienced pathologist evaluated the organ involvement and the IFI pathologic pattern. Fisher’s Exact test was used to recognize the IFI prevalence, the main occurring pathogens and the involved site; a p-value of <0.05 was considered statistically significant.

Results

The analysis of 161 consecutive autopsies identified 40  pts.(25%)resulting to have IFI; of these, 22 were AML (55%) ,6 ALL (15%),11LPD (28%) and 1 other. Aspergillus  spp. infection was detected in 20 cases (50%), Mucor spp in 8 (20%) and Candida spp. in 12 (30%). Moulds  were prevalent in acute leukemia pts. and Aspergillus spp. is the leading pathogen with respect to Candida and Mucor spp. (p 0,0396),with a statistically significant prevalence in ALL (p 0,0186).The site more involved resulted lung (p 0.0002). Whereas the standardized EORTC/MSG criteria applied in vivo were conclusive for  IFI in 6  pts ( 15%) only, the postmortem findings revealed fungal infections in further 34  pts (85%).

Conclusion

This analysis confirms that the IFI diagnosis is still an unresolved issue in hematological malignancies. Acute leukemias remain the subset with the higher prevalence of mould infections. As in other largest studies, in our experience Aspergillus spp and lung proved to be the most recurrent pathogen and site of involvement. At now, the diagnostic methods are not still completely able to identify the underlying IFI, thus  the autopsy rate should be increased to achieve a better knowledge of epidemiology and to critically review previous misdiagnosis.

Disclosures:

No relevant conflicts of interest to declare.
Synergism Between HOXA9 and Mutant JAK3 (M511I) Leads to Rapid Leukemia Development within an in Vivo Murine Bone Marrow Transplant Model
Authors: de Bock Charles E , Degryse Sandrine , Demeyer Sofie , Sweron Bram , Gielen Olga , Mentens Nicole , Geerdens Ellen , Cools Jan
Published: October 2019

Activation mutations in JAK3 occur in 16% of T-cell acute lymphoblastic leukemia (T-ALL) cases, and co-occur frequently with HOXA cluster rearrangement. Genomic rearrangement of the HOXA cluster results in increased expression of HOXA9 and HOXA10. However it remains unclear if either HOXA9 or HOXA10 can cooperate with activating JAK3 mutations during oncogenic transformation and leukemogenesis. We have previously shown that JAK3 mutations lead to cell transformation and cause a long latency T-ALL in vivo using a mouse bone marrow transplant model. In this study we demonstrate that co-expression of the activating JAK3(M511I) protein with HOXA9 cooperate to develop leukemia within 30 days of transplant using an in vivo bone marrow transplant model. In our cooperative model, murine hematopoietic stem / progenitor cells were co-transduced with either both retroviral vectors encoding JAK3(M511I)/GFP and HOXA9/mCherry or each individually and then injected into sub-lethally irradiated recipient mice. Mice transplanted with bone marrow cells expressing JAK3(M511I) mutant alone developed T-ALL in 120 to 150 days. In sharp contrast, mice transplanted with cells expressing both JAK3(M511I) and HOXA9 showed rapid leukemia development within 30 days after transplant. Leukemia development was characterized by the rapid and specific increase in GFP-mCherry double positive cells. These animals showed high WBC, and splenomegaly and accumulation of immature CD8 single positive cells in the thymus. Similar experiments with HOXA10 did not show cooperation suggesting that HOXA9 is the more important oncogene in HOXA rearranged leukemias when a JAK3 activating mutation is present. To determine the underlying genetic mechanism for cooperation between HOXA9 and JAK3(M511I) the single positive JAK3 and double positive JAK3/HOXA9 expressing cells were isolated from thymi of leukemic mice for both epigenomic profiling using ATAC-seq and gene expression profiling. These analyses identified genetic pathways activated by the co-expression of HOXA9 and JAK3(M511I) mutation and provide mechanistic insight into the synergistic interaction between these two factors in driving leukemia development. Treatment of the animals with a JAK kinase inhibitor resulted in delayed leukemia development, confirming that the leukemia cells remain sensitive to the JAK inhibitor. This mouse model provides insight in the cooperation between oncogenes in leukemia development and provides a model for the study of targeted agents in this setting.

Disclosures

No relevant conflicts of interest to declare.
Anthracycline Type during Induction Associated with Outcome in Pediatric t(8;21) and Inv(16) AML
Authors: Hasle Henrik , Abrahamsson Jonas , De Bont Evelina S , de Haas Valerie , De Moerloose Barbara , Forestier Erik , Ha Shau Yin , Heldrup Jesper , Jahnukainen Kirsi , Jónsson Ólafur G , Lausen Birgitte , Palle Josefine , Reedijk Ardine , Saks Kadri , Zeller Bernward
Published: October 2019

Background. The core-binding factor AMLs, t(8;21)(q22;q22), AML1-ETO or RUNX1-RUNX1T1 and inv(16)(p13q22), CBFB–MYH11, are considered as favorable risk factors in AML with superior event-free survival (EFS) and overall survival (OS). However, within the AML protocol from the Nordic society of pediatric hematology/oncology (NOPHO) from 2004 we observed a decrease in EFS for t(8;21) AML.

Material. We analyzed all children with t(8;21) or inv(16) treated on NOPHO-AML 93 and NOPHO-AML 2004 from Denmark, Finland, Iceland, Norway, and Sweden from 1993 to 2013 and patients from countries later joining the NOPHO-AML 2004 protocol; Estonia (2004), Hong Kong (2007), Belgium and the Netherlands (2010). The choice of anthracycline during induction changed from two courses including doxorubicin (75 mg/m2 in each course), Dox-Dox, in the NOPHO-AML 93 to idarubicin (36 mg/m2) and mitoxantrone (30 mg/m2), Ida-Mitox, in NOPHO-AML 04. After the observation of a decrease in EFS in patients with t(8;21) in late 2010 it was recommended to give fludarabine, cytarabine, and DaunoXome (liposomal daunorubicin 180 mg/m2) as second induction to patients with t(8;21); Ida-DNX.

Results. From 1993 to 2013 a total of 90 children, median age 9.1 years range 1-16, with t(8;21) AML were treated with Dox-Dox (n=25), Ida-Mitox (n=41), and Ida-DNX (n=24). Hematopoietic stem cell transplantation was given to 10 patients (11%), mostly with sibling donors in the NOPHO-AML 93 protocol (n=7). A reduction of the 3-year EFS was noted in patients treated with Ida-Mitox (41%) compared with Dox-Dox (64%). Those treated with Ida-DNX had significantly higher 3-year EFS of 87% (Figure). The events in t(8;21) AML on Ida-Mitox were relapses (n=20), death in CR1 (n=2), and secondary malignancy (n=2). The salvage rate for relapsed t(8;21) patients remained high and the overall favorable OS did not change significantly (Figure). From 1993 to 2013 a total of 58 children with inv(16) AML were treated with Dox-Dox (n=18) and Ida-Mitox (n=40). No significant changes in EFS (72% vs. 65%) or OS (89% vs. 79%) were observed for patients with inv(16).

Conclusion. Despite the overall favorable survival in t(8;21) the present study suggests that the choice of anthracycline during induction may be of major importance for the EFS in t(8;21) AML. Idarubicin and mitoxantrone during induction was associated with an inferior EFS compared with doxorubicin in t(8;21) AML. DaunoXome combined with fludarabine and cytarabine seemed to be particular effective during induction therapy in t(8;21) AML.

Figure 1 Figure 1.

Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
New Interleukin-15 Superagonist (IL-15 SA) Combined with Donor Lymphocyte Infusion (DLI) Significantly Enhances Graft-Versus-Tumor Activity after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Bailey Cavan , Panis Michelle M , Buyukgoz Cihangir , Budak-Alpdogan Tulin , Flomenberg Neal , Alpdogan Onder
Published: October 2019

Donor lymphocyte infusion (DLI) has been successfully used clinically to augment the graft-versus-tumor (GVT) effect following hematopoietic stem cell transplantation (HSCT) in relapsed patients. However, improvements can still be made in enhancing anti-tumor activity, reducing graft-versus-host disease (GVHD) and decreasing complications from opportunistic infections. Our studies present clear evidence of increased tumor clearance via cytokine therapy in combination with DLI as a way to “boost” the infused cells function. Interleukin-15 (IL-15) is a potent cytokine that increases CD8+ T and NK cells number and function in normal mice and recipients of stem cell transplantation. Despite this, obstacles remain for use of IL-15 therapeutically, specifically its low potency and short in vivo half-life. To overcome this, a new IL-15 superagonist (IL-15 SA-(ALT-803)) has been developed with a longer half-life and increased potency.

Administration of IL-15 SA to recipients of CFSE labeled T cells increases proliferation of CD8+T cells and IFN-γ and TNF-α secretion from CD8+T cells. We developed a murine DLI model by titrating the dose of infused T cells in a parent-F1 model, and then combined IL-15 SA administration with DLI in murine recipients of allogeneic HSCT. In this model, lethally irradiated CB6F1 (H2Kb/d) mice were transplanted with T- cell depleted bone marrow cells from C57BL6 mice (H2Kb). All recipients of HSCT were also co-injected A20 B-cell lymphoma cells transfected with a luciferase-producing gene, which allows bioluminescent imaging and tracking of tumor progress in vivo. Mice receiving DLI (2.5 X 105 T cells) with IL-15 SA injections given at 1μg/mouse on days 17 and 24 post-BMT show less tumor burden and increased overall survival (p = 0.04) and decreased tumor growth (p = 0.02) (Figure 1). The IL-15 SA treated group had a significantly less weight loss than the control group (p = 0.007). No GVHD symptoms were noted via weekly clinical scoring, highlighting both the efficacy and overall safety of the IL-15 SA therapy. Furthermore, we evaluated T- cell exhaustion markers on CD8+ T cells in surviving mice. We found increased programmed death-1 (PD-1) expression on T cells even when the tumor burden is cleared. Treatment with IL-15 SA reduced PD-1 expression on donor CD8+ T-cells in mice surviving more than 120 days post-transplant.

We conclude that IL-15 SA enhances CD8+ T cell function by increasing cytokine secretion and proliferation of T cells whereas could also prevent T cell exhaustion. We suggest that IL-15 SA is a long-waited lymphoid growth factor and has the potential to use in combination with DLI for the treatment of recurrent disease after HSCT.

Disclosures

No relevant conflicts of interest to declare.
FLT Imaging Reveals Kinetics and Biology of Engraftment after Myeloablative HSCT
Authors: Williams Kirsten M , Holter Chakrabarty Jennifer L. , Lindenberg Liza , Adler Steve , Gea-Banacloche Juan , Blacklock-Schuver Bazetta , Hakim Frances T. , Hickstein Dennis D. , Kochenderfer James N , Wilder Jennifer , Chinn Tracey , Kurdziel Karen , Steinberg Seth M. , Khuu Hanh , Lin Frank I , Fowler Daniel H. , Halverson David , Avila Daniele N. , Selby George , Taylor Tiffani N. , Mann Jennifer , Hsu Jennifer , Epstein Robert B. , Anderson Stacy L , Chuong Nguyen , Havlicek Joseph , Li Shibo , Pham Tom , Kraus Teresa , Vesely Sara K. , Pavletic Steven Z. , Bollard Catherine M. , Choyke Peter , Gress Ronald E
Published: October 2019

Bone marrow failure is associated with high morbidity and mortality after hematopoietic stem cell transplantation (HSCT) due to complications associated with prolonged neutropenia. Using the positron emission tomography (PET) imaging probe, 3’-deoxy-3 18F-fluorothymidine (18FLT), we illustrate HSC homing very early after myeloablative HSCT in adults in vivo. In our prospective trial, NCT01338987, we imaged 14 patients undergoing myeloablative transplantation with Cytoxan and total body irradiation and evaluated corresponding uptake of radiotracer 18FLT in osseous structures throughout the bone marrow compartment following full myeloblation, early engraftment, and hematopoietic reconstitution. 18FLT was safe and did not delay engraftment. Objective increases in 18FLT uptake measured by standardized uptake value (SUV) revealed evidence of stem cell homing to medullary bone marrow sites within 5 days of donor cell infusion. In patients without relapse (n=11), mean thoracic spine SUV was greater among patients with rapid (<21 days) as compared to slow engraftment (≥21 days). Using SUV, we could differentiate full ablation (SUV of 0.65) from early engraftment at day +5-6 (SUV 1.1, p <0.05). 18FLTsignal increases revealed a consistent pattern of marrow recovery that recapitulated fetal ontogeny, claiming sites of hematopoiesis that are normally dormant in adults. These findings were substantiated with in vitro cellular recovery data. 18FLT SUV could also predict marrow recovery after secondary graft failure without additional HSC infusion. In summary, our data show that 18FLT reveals homing, kinetics, and biology of hematopoietic engraftment in adults in vivo.18FLT could predict engraftment very early after HSCT and could be of value to predict primary or secondary graft failure in high risk populations, such as recipients of cord blood transplantation or haplo-identical transplantation.

Disclosures

No relevant conflicts of interest to declare.
Ethnic Disparities in Survival of Chronic Myeloid Leukemia
Authors: Ghimire Krishna B , Shah Binay K.
Published: October 2019

Introduction:

Development of tyrosine kinase inhibitors has led to significant improvement in survival of chronic myeloid leukemia (CML) patients. We conducted this study to evaluate any ethnic disparity in CML survival.

Methods:

We analyzed surveillance, Epidemiology, and End Results, SEER 18 registry to compare 1 and 3 years relative survival rates of CML patients diagnosed between 2001- 2008 by ethnicity - Caucasian, African American (AA), Others (O). We analyzed survival rates by age (≤65, >65), and time periods: (2001-2004) and (2005-2008). We used SEER*Stat software to calculate Z value.

Results:

There were 6,306 CML patients during (2001-2008); 3,111 during (2001-2004) and 3,195 during (2005-2008). The 1-year relative survival rates among Caucasians (n=6,306), AA (n=790), and Others (n=532) were 80.0±0.5%, 83.8 ±1.4%, and 84.0±1.7% [Z value 2.43 (AA vs Caucasian), 2.15 (O vs Caucasian)] respectively. The 3 years RS rates were 65.6±0.7%, 70.2±1.8%, and 70.9 ±2.1% respectively with Z value 2.49 (AA vs Caucasian) and 2.42 (Caucasian vs O).

Among younger patients (age ≤65 years, n=4,245) Caucasians had better RS compared to AA (83.1±0.7% vs 78.7±1.8%, Z value 2.24) at 3 years. There was no difference survival at 1and 3 years in older patients (age >65 years, n= 3,383).

The survival rates of patients diagnosed during 2001-2004 were similar for all ethnic groups. Among patients diagnosed during 2005-2008, survival rates were significantly higher for AA versus Caucasians (1 year RS 86.6±1.8% vs 81.3±0.7%, Z value 2.45, and 3 year RS 73.8±2.4% vs 68.3±0.9%, Z value 2.20). Others also had better RS compared to Caucasians (87.1±2.1% vs 81.3±0.7%, Z value 2.22) at 1 year.

There was no racial disparity in survival rates when analyzed by age (≤65 and >65) and sex at 1 and 3 years during 2001-2004 and 2005-2008.

Conclusions:

Our study showed that there is ethnic disparity in CML survival. Among CML patients diagnosed during 2005-2008, AA and Other races had superior survival rates compared to Caucasians.

Disclosures

No relevant conflicts of interest to declare.
Prognostic Understanding, Quality of Life, and Mood in Patients Undergoing Hematopoietic Stem Cell Transplantation
Authors: El-Jawahri Areej , Traeger Lara , Kuzmuk Kailyn , Eusebio Justin , Vandusen Harry , Shin Jennifer , Gallagher Emily , Greer Joseph , Pirl William , Jackson Vicki , Ballen Karen K. , Spitzer Thomas R. , Graubert Timothy A. , McAfee Steven L. , Dey Bimalangshu R. , Chen Yi-Bin , Temel Jennifer S.
Published: October 2019

Background: In advanced cancers, patients' prognostic perceptions influence their medical decisions. However, little is known about how patients with hematologic malignancies undergoing hematopoietic stem cell transplantation (HCT) and their family caregivers (FC) understand their prognosis. We examined prognostic understanding in these patients and their FC during hospitalization for HCT and its relationship with quality of life (QOL) and mood.

Methods: We conducted a longitudinal study of patients hospitalized for HCT and their FC. At baseline (6 days pre-HCT), we used a 10-item questionnaire to measure patients' and FC information preferences and perception of prognosis. We also asked oncologists to estimate the patients' prognosis. At day-6, day+1, day+8 of HCT, we assessed QOL (Functional Assessment of Cancer Therapy-Bone Marrow Transplantation (FACT-BMT)), and mood (Hospital Anxiety and Depression Scale). We evaluated the relationship between participants' prognostic understanding and their QOL and mood during hospitalization for HCT using multivariable linear mixed models.

Results: We enrolled 97% of consecutively eligible patients undergoing autologous (n=30), myeloablative (n=30) or reduced intensity (n=30) allogeneic HCT. Most patients (80/90, 88.9%) and FC (41/47, 87.1%) believed that it is extremely or very important to know about prognosis. However, the majority of patients (66/85, 77.6%) and FC (33/46, 71.7%) reported an inaccurate and more optimistic perception of the patients' prognosis compared to the oncologist (P < 0.0001) [Figure 1]. Patients with an accurate understanding of their prognosis reported worse QOL (F = 4.3, P = 0.03) and a higher rate of depression (F = 4.4, P = 0.04) with worsening depressive symptoms over time (F = 4.13, P = 0.006) compared to patients with an inaccurate prognostic perception.

Conclusions: The majority of patients and FC reported inaccurate and more optimistic perception of prognosis compared with the oncologist. Patients with an accurate perception of their prognosis had lower QOL, higher rates of depression, and worsening depression symptoms during hospitalization for HCT. Interventions are needed to improve prognostic understanding, while providing them with the psychosocial support they need to cope with this difficult information.

Figure 1: Patients', Family Caregivers', and Oncologists' Prognostic Perception Figure 1:. Patients', Family Caregivers', and Oncologists' Prognostic Perception

Disclosures

Off Label Use: Sorafenib for FLT3-ITD AML.
Development of Quality Metrics to Evaluate Pediatric Hematologic Oncology Care in the Outpatient Setting
Authors: Teichman Jennifer R , Gupta Sumit , Punnett Angela
Published: October 2019

Introduction:

Systems to quantify and incentivize quality of care (QoC) have been developed in multiple healthcare settings. In pediatric oncology, lists of QoC metrics or recommendations have been procured through consensus methodologies such as the Delphi process. To date, no QoC metrics have been developed for outpatient pediatric oncology.

Objectives:

The aim of this study was to develop a list of QoC metrics for the leukeumia-lymphoma (LL) clinic at the Hospital for Sick Children in Toronto, using a consensus process that could be adapted to other clinic settings.

Methods:

A modified Delphi process following the American Society of Clinical Oncology (ASCO) guidelines was used to generate consensus on a list of QoC metrics (Loblaw et al., 2012). A Medline-Ovid search was conducted for quality indicators, metrics and recommendations relevant to pediatric oncology. Results were screened for (a) system-level metrics that could be translated to a clinic level and (b) clinic-level recommendations that could be converted to measurable quantities. Additional metrics outside the literature search were considered. A provisional list was compiled and circulated electronically to local stakeholders, including medical and nursing staff (n=10). Stakeholders ranked each metric on a 5-point Likert scale based on importance and feasibility of measurement (round 1). Stakeholders provided feedback on the metrics and suggested additional metrics. Median, interquartile range and full ranges were calculated for each metric. A metric was considered to reach consensus if the percent of respondents ranking within two consecutive scores was ≥70%. Results and comments from round 1 were re-circulated to stakeholders in personalized reports. This allowed each stakeholder to compare his or her previous scores with overall scores for each metric. Stakeholders were asked to re-rank each metric (round 2).

Results:

The literature search yielded 2 relevant publications from which a provisional list of 27 metrics was generated. Metrics were grouped into 7 categories (Table 1). In round 1, 19/27 (70%) metrics reached consensus. Stakeholders’ comments resulted in 4 new metrics and edits to 8 original metrics. All metrics were included in round 2 for a total of 31. Twenty-four of 31 (77%) metrics reached consensus after round 2 (Table 1). Thirteen were chosen for the final list based on highest consensus scores, highest interquartile and full ranges, and minimizing redundancy.

Conclusion:

This study demonstrates the feasibility of using a modified Delphi process to generate QoC metrics for a pediatric hematology oncology clinic, and provides a model other clinics may employ for local use. The final metrics will be used to evaluate the quality of care in the LL clinic, and to identify areas for improvement in clinic function.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Non-Coding RNAs Transcribed from ERV-9 LTR Retrotransposons Regulate Erythropoiesis
Authors: Hu Tianxiang , Pi Wenhu , Tuan Dorothy
Published: October 2019

Long noncoding RNAs (lncRNAs) regulate diverse cellular processes in development, differentiation and malignancy. In human cells, over 80% of the lncRNAs contain retrotransposon sequences transcribed from Alu, L1 and LTR retrotransposons, which comprise ~40% of the human genome. The functional significance of the retrotransposon lncRNAs is largely unknown. The human genome contains ~4000 copies of the ERV-9 LTR retrotransposon, which exhibits strong enhancer activity and initiates synthesis of ERV-9 lncRNAs in erythroid progenitor cells. Recently, we discovered that depletion of the ERV-9 lncRNAs in human erythroid progenitor cells cultured ex vivo from peripheral blood CD34+ cells inhibited ex vivo erythropoiesis. Whole genome RNA sequencing (RNA-seq) found that depletion of ERV-9 lncRNAs significantly suppressed transcription of 608 genes including ~50 key erythroid genes. We hypothesize that the ERV-9 lncRNAs together with the ERV-9 LTR act in cis to regulate transcription of these key erythroid genes and other genes to set up a transcriptional network that promotes erythropoiesis.

In the human b-globin gene locus, we showed previously that the ERV-9 LTR retrotransposon performs a beneficial biological function: The ERV-9 LTR enhancer binds NF-Y and GATA-1 and -2 to assemble an LTR-pol II transcription complex, which transcribes long, noncoding RNAs (lncRNAs) from the LTR R-U5 regions through the downstream locus control region (LCR) and intergenic DNAs to reach and activate transcription of b-globin gene 70 kb away. In this tracking and transcription (T&T) mechanism of long-range LTR enhancer function, the ERV-9 lncRNAs could be merely by-products of the tracking and transcribing process of the LTR complex without any functional significance. However, we found recently that depletion of the ERV-9 lncRNAs suppressed transcription of the entire human globin gene locus, diminished occupancies of NF-Y, GATA-1 and -2 and pol II at the ERV-9 LTR and reduced the looping frequency of the ERV-9 LTR with the globin gene locus in erythroid progenitor cells. Thus, the ERV-9 lncRNAs acted in cis to facilitate assembly of the LTR-pol II complex and modulate long-range LTR enhancer function in transcriptional activation of b-globin gene. Genome-wide, whether the ERV-9 lncRNAs transcribed from other gene loci perform similar biological function is under investigation.

Disclosures

No relevant conflicts of interest to declare.
Elevated Red Cell Reduced Glutathione Is a Novel Biomarker of Diamond-Blackfan Anemia
Authors: Utsugisawa Taiju , Uchiyama Toshitaka , Ogura Hiromi , Aoki Takako , Hamaguchi Isao , Ishiguro Akira , Ohara Akira , Kojima Seiji , Ohga Shouichi , Ito Etsuro , Kanno Hitoshi
Published: October 2019

Diamond-Blackfan anemia (DBA) is a rare congenital red cell aplasia characterized by congenital anomalies and predisposition to cancer. Recent observation disclosed that heterogeneous mutations in ribosomal protein (RP) genes are present in approximately 50% of patients, suggesting that diagnosis should be made by clinical phenotypes such as age, hematological findings or positive family history. Although elevated activity of red cell adenosine deaminase (eADA) has been utilized as a useful biomarker for differential diagnosis of DBA, approximately 20% of DBA patients are eADA-negative. Recent observations suggested that ribosomal haploinsufficiency increases oxidative stress, leading to p53 gene activation and premature death of erythroid cells. We hypothesized that reduced glutathione (GSH), an essential antioxidant of erythroid cells, might be upregulated in red cells of DBA subjects. In order to test this hypothesis, we examined red cell GSH as well as eADA of 22 patients in 18 DBA families, in whom we had identified gene mutations in RPS19, RPL5, RPL11, RPS10, RPS17 or RPS35a. All except one DBA patients showed elevated GSH (>88.6 mg/dl RBC, M+SD), whereas 17 out of 22 patients exhibited elevated eADA (>2.31 IU/g Hb, M+3SD). We also examined 14 unaffected members of the DBA families, with 1 out of 14 subjects showing elevated GSH and none showing elevated eADA. We performed linear discriminant analysis between DBA and non-DBA subjects with both eADA and GSH using the Support Vector Machine (SVM) from 36 subjects, and successfully obtained a formula to discriminate DBA from unaffected subjects: 0.937*eADA+0.0702*GSH-7.9044 >0. By using this formula, all DBA examined can be diagnosed and unaffected family members can be excluded. Since approximately 50% of clinically diagnosed DBA cases have no causative RP gene mutations, the combined assessment of eADA and GSH might be quite useful for biochemical diagnosis of DBA.

Disclosures

No relevant conflicts of interest to declare.
An Unsuspected Dyserythropoiesis in Gaucher Disease
Authors: Reihani Niloofar , Arlet Jean-Benoît , Billette de Villemeur Thierry , Belmatoug Nadia , Colin Yves , Hermine Olivier , Le Van Kim Caroline , Franco Mélanie
Published: October 2019

Introduction: Gaucher disease (GD) is a lysosomal storage disorder, impairing glucosylceramide catabolism. GlcCer-laden macrophages transform into Gaucher cells and are responsible for the major GD symptoms (tissue infiltration, organomegaly, bone lesion). However, other signs of the disease such as anemia, spleen and bone infarcts might involve red blood cells (RBC). Despite the lysosomal characteristic of GD and the evidence of the central rdole of macrophages, our recent findings pointed out unexpected RBC abnormalities (Franco et al, Blood. 2013;121:546-555). These findings uncovered an overlooked aspect in GD showing that GD RBC exhibit abnormal rheological, morphological and membrane adhesion properties. Thus, RBC could be considered as culprit for ischemic events in GD. In this study, we analyzed the erythroid differentiation in GD to address the etiology of red cell abnormal behavior. We induced in vitro erythropoiesis from circulating GD patient progenitors.

Methods: After purification of CD34+ cells from peripheral blood of Gaucher patients (n=20) or controls (CTL), erythroid progenitors were expanded in the absence of erythropoietin (EPO) and differentiated in a second step with erythropoietin. GD and CTL CD34- cells were differentiated into macrophages by addition of M-CSF in the culture medium. To determine the effect of CTL and GD macrophages on normal erythropoiesis, in vitro generated macrophages were co-cultured with CTL erythroid progenitors in the presence of EPO. Proliferation of progenitors as well as erythroid differentiation were examined by flow cytometry and cytology.

Results: Our preliminary results showed: (i) a dramatic decrease in proliferation of progenitors in the first step of expansion (**p <0.005), (ii) an accelerated erythroid maturation as evaluated by the percentage of cells expressing and non-expressing Glycophorin A and C-Kit, respectively (**p <0.005), (iii) an absence of effect of Gaucher macrophages co-culture on normal erythroid terminal maturation, and (iv) a correlation between the acceleration level of maturation and the degree of anemia in GD patients as determined by hemoglobin level (**p<0.005).

Conclusion: We evidenced an unexpected dyserythropoiesis characterized by accelerated erythroid differentiation in GD. The observed dyserythropoiesis is independent of macrophage defects and is intrinsic to the GD erythroid lineage. These studies by helping to understand the role of erythroid cells in the pathophysiology of GD as well as the mechanism of anemia should prove useful for the development of additive therapies to alleviate debilitating clinical complications of this morbid disease such as osteonecrosis and splenomegaly.

Disclosures

Le Van Kim: SHIRE: Research Funding.
Edx-17: A Novel, Safe and Efficacious Treatment for Sickle Cell Disease
Authors: Broyles Robert H. , Joshi Sunil K. , Curtis Carol D. , Roth Austin C. , Floyd Patrick A. , Trudel Marie , Belegu Visar , Baker Ashley A. , Floyd Robert A..
Published: October 2019

Gene regulation of developmental hemoglobin switching holds the potential for therapeutic relief from all symptoms associated with Sickle Cell Disease (SCD). Reactivation of fetal gamma-globin expression (HbF) can replace mutant betaS-globin (HbS) to produce functional hemoglobin tetramers and eliminate the hemoglobin polymerization that is characteristic of sickled red blood cells. We have discovered a protein that regulates this developmental switch, and have identified a compound that stimulates expression of this protein. EdX-17 promotes expression of the anti-stress factor ferritin heavy chain (FtH), which enters the nucleus of erythroid precursor cells and activates expression of fetal gamma-globin, producing HbF (PNAS 98:9145-50, 2001; Blood 108:790a, 2006). Mononuclear cells were isolated from SCD patient blood and matured in vitro to the advanced erythroblast stage using a 28-day, 2-phase culture system (Methods in Molecular Biology 482:127-40, 2009; Blood 119:6296-306, 2012). Treatment with EdX-17 for 24h resulted in a dose-responsive induction of gamma-globin gene expression and a concomitant dose-responsive increase in HbF was observed after 28 days in culture. These studies demonstrate that EdX-17 doses in the picomolar range are sufficient to significantly enhance HbF. Furthermore, EdX-17 treatment reconstitutes fetal hemoglobin (HbF) in transgenic betaYAC mice to levels above 25-30% - the range thought to be sufficient to ameliorate symptoms of SCD – with no detectable ill effects. In fact, mice treated with EdX-17 tend to have shinier coats, are more alert and stronger than age-matched, untreated mice. Development of this novel therapeutic is expected to ameliorate SCD symptoms, decrease pain and morbidity, increase life-span, greatly improve patient quality of life, and significantly reduce treatment costs. Supported in part by The Sickle Cell Cure Foundation, Inc., the Bill & Melinda Gates Foundation, and EpimedX, LLC.

Disclosures

Broyles: EpimedX, LLC: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding. Curtis:EpimedX, LLC: Employment, Equity Ownership, Research Funding. Roth:EpimedX, LLC: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding. Floyd:EpimedX, LLC: Employment, Equity Ownership, Research Funding. Belegu:EpimedX, LLC: Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding. Floyd:EpimedX, LLC: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding.
Inorganic Polyphosphate Activates mTOR Complexes 1 and 2 in Vascular Endothelial Cells
Authors: Hassanian Mehr Seyed Mahdi , Dinarvand Peyman , Rezaie Alireza R.
Published: October 2019

Inorganic polyphosphate (polyP) elicits proinflammatory signaling responses in endothelial cells through interaction with two receptors, RAGE and P2Y1. We demonstrate here that polyP elicits proinflammatory signaling responses through activation of mTOR complexes 1 (mTORC1) and 2 (mTORC2) in endothelial cells. Thus, polyP effectively induced the phosphorylation of the mTORC1 substrate, p70S6K, in endothelial cells by an AKT-dependent but ERK-independent mechanism. Furthermore, it was found that activation of the PI3K/PLC/PKC/Ca2+ signaling axis is required for the polyP-mediated phosphorylation of p70S6K. The inhibition of mTORC1 by either rapamycin or by the siRNA knockdown of raptor (mTORC1-specific component) but not rictor (mTORC2-specific component) abrogated polyP-mediated phosphorylation of p70S6K. By contrast, the siRNA knockdown of rictor but not raptor eliminated the barrier disruptive effect of polyP, suggesting that the barrier permeability regulating function of polyP is mediated through activation of mTORC2. Specific NF-kB inhibitors abrogated polyP-mediated phosphorylation of p70S6K and rapamycin suppressed polyP-induced activation of NF-kB, suggesting both proinflammatory and mTOR signaling functions of polyP are linked. Finally, inhibitors of mTOR or its upstream signaling network eliminated polyP-mediated vascular leakage and leukocyte recruitment in animal models, suggesting that findings in cellular models are physiologically relevant.

Disclosures

No relevant conflicts of interest to declare.