bluuebunny/crossref_metadata_2025 huggingface dataset:
G-CSF-Primed Bone Marrow As a Source of Hematopoietic Stem Cells for Allogeneic Transplantation in Malignant and Non-Malignant Hematological Disorders
Authors: León-Rodríguez Eucario , Guzmán-Uribe Patricia , Deffis Court Marcela , Perez-Alvarez Sandra Ileana , Rivera Franco Monica M
Published: November 2019

Introduction: To date, worldwide, the main source of stem cells is peripheral blood (PBSCs). However, its use has been associated with a higher incidence of graft versus host disease (GVHD) when compared with the use of bone marrow. It has been demonstrated that the fast engraftment using G-CSF-primed bone marrow as a source of stem cells, compares to that seen with PBSCs, and it is also associated with a decreased incidence of GVHD.

Objective: To describe the results obtained from allogeneic, G-CSF-primed bone marrow transplantations, at INCMNSZ, from November 1998 to March 2013.

Material and methods: A retrospective analysis was performed in patients who underwent allogeneic, G-CSF-primed bone marrow transplantation. Clinical characteristics, frequency of GVHD, and survival, were conducted, using the Statistical Software Package SPSS v21.0.

Results: Forty-nine patients who underwent allogeneic, G-CSF-primed bone marrow transplantation, from November 1998 to March 2013, were included. Patients (male, 63%) had a median age of 29 years (range 16-59). The patients had a following range of underlying diseases: aplastic anemia (n=15, 30.6%), myelodysplastic syndrome (n=12, 24.5%), chronic myeloid leukemia (CML, n=9, 18.4%), acute lymphoblastic leukemia (LLA, n=7, 14.3%), acute myeloid leukemia (AML, n=3, 6.1%), or others (n=3, 6.1%). Patients achieved a bilineage engraftment with a median time of 20 days (range, 11-40) for absolute neutrophil count and 15 days (range, 5-45) for platelets. Acute GVHD was observed in 4 patients (8.2%), reported as grade I and II; and 14 patients (28.6%) experienced a limited form of chronic GVHD. Nine out of 49 (18.3%) patients relapsed: CML (n=5), AML (n=1), AA (n=1), ALL (n=1), others (n=1). All patients with relapsed CML were treated with donor lymphocyte infusions, but only 2 responded. Treatment related mortality (TRM) was 8.16%, including infectious complications in 2 patients, cGVHD in one patient, and veno-oclussive disease (VOD) in one patient. Overall mortality was 26.5%; the majority of deaths (69.2%) were caused by progressive and relapsing disease. With a follow up of 43 months (0-149), the median overall survival (OS) has not been reached; however, the estimated 10-year OS is 69%.

Conclusion: According to our data, it seems that G-CSF-primed bone marrow is a better source of hematopoietic stem cells for allogeneic transplantation in malignant and non-malignant hematological disorders, when compared with PBSCs, due to the similar engraftment time, and a decreased incidence and severity of acute and chronic GVHD. Nevertheless, this strategy does not appear to be valid as an approach for diseases were graft versus tumor effect (GVT) is important to eradicate the malignancy, such as in CML.

Disclosures

No relevant conflicts of interest to declare.
IP3 3-Kinase B Controls Hematopoietic Stem Cell Homeostasis and Prevents Lethal Hematopoietic Failure in Mice
Authors: Sauer Karsten
Published: November 2019

Tight regulation of hematopoietic stem cell (HSC) homeostasis ensures life-long hematopoiesis and prevents blood cancers. The mechanisms balancing HSC quiescence with expansion and differentiation into hematopoietic progenitors are incompletely understood. Here, we identify inositoltrisphosphate (IP3) 3-kinase B (Itpkb) as a novel essential regulator of HSC quiescence and function. Young Itpkb-/- mice accumulated phenotypic HSC which were less quiescent and proliferated more than wildtype controls. Itpkb-/- HSC downregulated quiescence and stemness associated mRNAs, but upregulated activation, oxidative metabolism, protein synthesis and lineage associated transcripts. Although they showed no significant homing defects and had normal to elevated viability, Itpkb-/- HSC had a severely reduced competitive long-term repopulating potential. Aging Itpkb-/- mice lost hematopoietic stem and progenitor cells and died with severe anemia. Wildtype HSC normally repopulated Itpkb-/- hosts, indicating a HSC-intrinsic Itpkb requirement. In vitro, Itpkb-/- HSC had reduced cobblestone-area forming cell activity and showed increased stem cell factor activation of the phosphoinositide 3-kinase (PI3K) effector Akt. This was reversed by exogenous provision of the Itpkb product IP4, a known PI3K/Akt antagonist. Itpkb-/- HSC also showed transcriptome changes consistent with hyperactive Akt/mTOR signaling. Thus, we propose that Itpkb ensures HSC quiescence and function in part by limiting cytokine-induced PI3K signaling in HSC.

Disclosures

No relevant conflicts of interest to declare.
Comparable Leukemia-Free Survival after Autologous Hematopoietic Cell Transplantation Versus HLA-Identical Sibling Hematopoietic Cell Transplantation for Adult Acute Myeloid Leukemia in First Complete Remission: A Registry Study By the Adult AML Working Group of the Japan Society for Hematopoietic Cell Transplantation
Authors: Mizutani Motonori , Hara Masahiko , Fujita Hiroyuki , Aoki Jun , Kanamori Heiwa , Machida Shinichiro , Yamasaki Satoshi , Ohashi Kazuteru , Usuki Kensuke , Fukuda Takahiro , Chou Takaaki , Tanaka Junji , Atsuta Yoshiko , Takami Akiyoshi
Published: November 2019

Allogeneic hematopoietic cell transplantation (HCT) from a matched sibling donor (MSD) may provide a cure for acute myeloid leukemia in first complete remission (AML/CR1), although the procedure is associated with a higher rate of treatment-related mortality (TRM) than autologous HCT, and it remains uncertain which modality is preferable as post-remission treatment. We therefore conducted a retrospective registry-based analysis to compare the outcomes of patients with AML/CR1 receiving autologous peripheral blood (PB) grafts (n=398, median age: 47, range: 17-80) and those receiving allogeneic MSD bone marrow (BM) grafts (n=633, median age: 38, range: 16-73) or allogeneic MSD PB grafts (n=475, median age: 42, range: 16-74) between 1995 and 2011. Consequently, the 5-year overall survival (OS) rates for the autologous PB, allogeneic BM and allogeneic PB recipients were 62% (95% confidence interval [CI], 57-67%), 61% (95% CI, 57-65%; P=0.90) and 54% (95% CI, 49-59%; P=0.07), respectively (Fig. 1-A), and the 5-year leukemia-free survival (LFS) rates were 57% (95% CI, 52-62%), 58% (95% CI, 54-63%; P=0.49) and 51% (95% CI, 46-56%; P=0.12), respectively(Fig. 1-B). Meanwhile, the 5-year cumulative incidence of TRM was 8% (95% CI, 5-11%), 16% (95% CI, 13-19%; P=0.009) and 19% (95% CI, 15-23%; P=0.0001), respectively, while that of relapse was 35% (95% CI, 30-40%), 26% (95% CI, 22-29%; P=0.003) and 30% (95% CI, 26-35%; P=0.08), respectively. A multivariate analysis performed with autologous PB HCT as the reference showed a hazard ratio (HR) for OS of 0.93 (95% CI, 0.73-1.18; P=0.53) for allogeneic BM HCT and 1.08 (95% CI, 0.83-1.39; P=0.57) for allogeneic PB HCT and an HR for LFS of 0.86 (95% CI, 0.69-1.09; P=0.21) and 0.98 (95% CI, 0.77-1.24; P=0.85), respectively. Stratifying the patients according to cytogenetics (favorable, intermediate and poor) and age (<50 years and ≥ 50 years) did not influence the results. Accordingly, autologous PB HCT may be a viable alternative as post-remission therapy in patients with AML/CR1 in the absence of MSD.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Evaluation of the Disease Risk Index on Outcomes Following Allogeneic Hematopoietic Cell Transplantation for Acute Myeloid Leukemia and Myelodysplastic Syndromes
Authors: Michelis Fotios V. , Messner Hans A. , Uhm Jieun , Alam Naheed , Loach David , McGillis Laura , Lambie Anna , Gupta Vikas , Kuruvilla John , Lipton Jeffrey H. , Seftel Matthew D. , Kim Dennis D.
Published: November 2019

Allogeneic hematopoietic cell transplantation (HCT) is a curative treatment option when indicated for myeloid malignancies such as acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). The Disease Risk Index (DRI, Armand et al, 2012) has been developed to predict overall survival and progression-free survival on the basis of differences in relapse risk. The purpose of this single-center study was to retrospectively investigate the prognostic value of the DRI on the outcome of 470 patients that underwent HCT for AML and MDS between 2000 and 2013.

AML patients (n=381) underwent HCT in first and second complete remission and MDS patients (n=89) underwent HCT untreated or in remission. Median age at HCT was 51 (range 18-71), 219 (47%) patients were female. Myeloablative conditioning (MAC) was used in 304 (65%) patients, reduced-intensity (RIC) in 166 (35%) patients. Donors were related for 287 (61%) patients, unrelated for 183 (39%) patients. Grafts were peripheral blood stem cells (PBSC) in 377 (80%) patients and bone marrow in 93 (20%) patients. Median follow-up of patients alive was 44 months (range 1-134). In accordance with the DRI criteria, all 470 patients were in the low risk group concerning disease stage. Concerning disease biology characterized by cytogenetic risk, 11 patients were low risk, 396 patients were intermediate risk and 63 patients high risk. Based on the DRI overall risk stratification, 11, 396 and 63 patients were low, intermediate and high risk respectively.

Univariate analysis demonstrated that the DRI was significantly prognostic for overall survival (OS) with a 3-year OS of 82%, 48% and 29% for low, intermediate and high risk patients respectively (p=0.005, Figure A). For cumulative incidence of relapse (CIR), DRI was again prognostic with 3-year CIR 0%, 21% and 30% for low, intermediate and high risk patients respectively (p<0.0001, Figure B). For non-relapse mortality (NRM), DRI did not demonstrate significant prognostic relevance with 3-year NRM 18%, 32% and 43% for low, intermediate and high risk patients respectively (p=0.14).

Multivariable analysis for OS confirmed the prognostic significance of the DRI with hazard ratio (HR) 3.3 and 4.9 for intermediate and high risk respectively compared to low risk (p=0.01). For CIR, DRI was highly significant (p<0.0001) while for NRM, the DRI was not predictive (p=0.22). This study confirms the prognostic relevance of the DRI for OS and CIR in our cohort of patients undergoing HCT for AML and MDS.

Figure 1 Figure 1.

Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
Outcomes of Allogeneic Hematopoietic Stem Cell Transplantation in Children and Young Adults with Chronic Myeloid Leukemia: A CIBMTR Cohort Analysis
Authors: Chaudhury Sonali , HiJiya Nobuko , Hu Zhenhuan , Sparapani Rodney , Kalaycio Matt E. , Alyea Edwin P. , Popat Uday R. , Saber Wael
Published: November 2019

Chronic myeloid leukemia (CML) in pediatrics is rare accounting for 2–3% of leukemias. Data on allogeneic hematopoietic cell transplantation (HCT) in this group is scarce. Since the introduction of tyrosine kinase inhibitors (TKI), HCT in patients with CML especially with early disease has decreased. Children and young adults have long life expectancies, lower morbidity with HCT and are likely to recieve prolonged TKI therapy with increased risk for complications. HCT for early disease may still benefit this population.

We retrospectively evaluated outcomes in CML patients receiving myeoloablative HCT reported to the CIBMTR, comparing pediatric patients (n= 212) to young adults (n=200). Table 1 describes patient variables.

5 y overall survival (OS) and leukemia free survival (LFS) in <18 y was 71% (95%CI 65-77) and 51% (95%CI 43-58) respectively. In adjusted analysis, there was a statistically insignificant effect of prior TKI therapy on LFS (P=0.07), OS (0.06), transplant related mortality (TRM) (P=0.47) or relapse (P=0.88). Favorable factors for OS were early disease, matched sibling donors (MSD), bone marrow (BM) or cord blood (CB) grafts. LFS was superior with total body irradiation (TBI), non alemtuzumab based regimens and recent (>2006) HCTs. 5 year OS and LFS for the 18-25y group was 71 (95%CI 65-77)% and 53 (45-60)% respectively. In adjusted analysis there was no difference in OS (P=0.7), LFS (P=0.4) for 18-25y group compared to the pediatric group. Favorable factors for OS/DFS remained early disease, MSD and BM grafts.

Allogeneic HCT outcomes were comparable in children and young adults. HCT should remain an option for young patients with CML especially with a MSD using TBI containing regimens with BM as the stem cell source. We could not show an advantage for TKI use prior to HCT. Long term toxicities of TKI and allogeneic HCT need to be further evaluated in this group.

Table1 Description of variables for Allogeneic HCT in CML Table1. Description of variables for Allogeneic HCT in CML

Figure 1 Adjusted OS and LFS for <18y vs. 18-25y Figure 1. Adjusted OS and LFS for <18y vs. 18-25y

Disclosures

HiJiya: Pfizer: Consultancy; Jazz: Consultancy; Enzon: Consultancy; Sanofi: Consultancy.
Dysregulation of the TGF β Pathway in Induced Pluripotent Stem Cells (iPSCs) Generated from Patients with Diamond Blackfan Anemia (DBA)
Authors: Ge Jingping , Apicella Marisa , Mills Jason A. , Garcon Loic , French Deborah L. , Weiss Mitchell J. , Bessler Monica , Mason Philip J
Published: November 2019

DBA is an inherited bone marrow failure syndrome that usually develops in the first year of life with clinical features of red cell aplasia and sometimes variable developmental abnormalities. Most affected patients have heterozygous loss of function mutations of one of the 11 ribosomal proteins (RPs) or mutations in the GATA1 gene which encodes an erythroid specific transcription factor. We have previously demonstrated that induced pluripotent stem cells (iPSCs) from fibroblast of DBA patients with RPS19 or RPL5 mutations recapitulate the pathogenesis of DBA, with the mutant lines showing abnormal ribosome biogenesis and altered erythropoiesis. The mechanism whereby haploinsufficiency for RPs causes failure of erythropoiesis and the other DBA features is still unknown. We investigated the pathways that are affected in these DBA iPSCs using an Affymetrix human exon array, and we observed the striking dysregulation of the TGF β pathway in DBA lines. The TGF β downstream target genes, such as DKK1, BAMBI, FN1, COL3A1, COLA1A1 and PAI-1 significantly increased in the DBA iPSCs. The TGF β signaling is complex and can occur via a canonical pathway or by a number of non-canonical pathways. We measured levels of a number of intermediates in these pathways by western blot, and observed a significant increase in the levels of p-JNK, a mediator of a non-canonical pathway, in the DBA iPSCs. Moreover, when the mutant cells were corrected by ectopic expression of WT RPS19 or RPL5, levels of p-JNK returned to normal. We also investigated the SMAD family, which are mediators of the TGF β canonical pathway and are known to negatively regulate the regeneration of hematopoietic stem cells. We observed a drastic decrease in SMAD4, but no change in p-SMAD2. Again corrected lines showed normal expression levels of SMAD4. Our data suggests that the activation of a non-canonical TGF β pathway in the DBA iPSCs may lead increased expression of the downstream genes; and the decrease of anti-proliferative factor SMAD4 may explain how DBA iPSCs maintain their growth. We conclude that the mutations of RPS19 or RPL5 both affect ribosome biogenesis and TGF β signaling, which can cause the failure of erythropoiesis at the stem cell stage. We further suggest that the suppression of SMAD4 may be used as a therapeutic target for DBA treatment.

Disclosures

No relevant conflicts of interest to declare.
Allogeneic Hematopoietic Stem Cell Transplantation for Patients with Acute Myeloid Leukemia Transformed from Ph-Negative Myeloproliferative Neoplasm: A Study from the Adult AML Working Group of the Japan Society for Hematopoietic Cell Transplantation (JSHCT)
Authors: Takagi Shinsuke , Masuoka Kazuhiro , Uchida Naoyuki , Kurokawa Mineo , Nakamae Hirohisa , Tsudo Mitsuru , Iwato Koji , Ichinohe Tatsuo , Atsuta Yoshiko , Takami Akiyoshi
Published: November 2019

BACKGROUND/METHODS: Long-term survivors with Philadelphia chromosome-negative myeloproliferative neoplasms (Ph-neg MPNs) can eventually develop acute myeloid leukemia (AML, i.e. blast transformation). Allogeneic hematopoietic stem cell transplantation (allo-HSCT) can be the only curative treatment for such patients. To clarify its outcome, we retrospectively studied the cases with AML transformed from Ph-neg MPNs using the national registry data of JSHCT. The cases transplanted after 2000 were collected.

RESULTS: Thirty-nine cases were extracted (male, n=21; female, n=18). Median age was 57 years (range, 22-71). Underlying Ph-neg MPNs included essential thrombocythemia (ET, n=21), primary myelofibrosis (PMF, n=11) and polycythemia vera (PV, n=7). FAB classification was M0 (n=4), M1 (n=4), M2 (n=10), M3 (n=0), M4 (n=1), M5 (n=3), M6 (n=0), M7 (n=1) and unknown (n=16). Median value of WBC at diagnosis of AML was 8300/uL (250-338000). Karyotype at diagnosis of AML was normal (n=6), complex (n=12), others (n=17) and unknown (n=4). Thirty-two cases (82%) were not in remission at the time of allo-HSCT (1st relapse, n=7; primary induction failure, n=18; untreated, n=12). Median duration between diagnosis of AML and allo-HSCT was 134 days (range, 24-369). The donors were related bone marrow or related mobilized peripheral blood stem cell (rBM/rPBSC, n=8), unrelated bone marrow (uBM, n=15) and unrelated umbilical cord blood (uCB, n=16). Myeloablative conditioning regimens (MAC) were used in 15 cases, and the remaining 24 cases were conditioned by reduced-intensity regimens (RIC), according to CIBMTR definition (Giralt S, et al, 2009). Cumulative incidence of neutrophil engraftment was 74.4% at day 60 (patients engrafted, n=29; death before day 60, n=6; relapse before day 60, n=4). At 2 years after transplant, overall survival (OS) was 29.2%. The median duration of follow up of survivors was 1989.5 days (range, 285-3270). In univariate analysis, age (>57 vs. <57, p=0.87), underlying MPNs (PMF vs. ET vs. PV, p=0.16), disease status at allo-HSCT (CR vs. non-CR, p=0.09), conditioning regimen (MAC vs. RIC, p=0.95) and donor selection (rBM/PBSC vs. uBM vs. uCB, p=0.10) had no impact on OS. The only variable that influenced on OS was chromosome at diagnosis of AML (normal vs. others vs. complex, p=0.02). The cumulative incidence of relapse and non-relapse mortality at 3 years was 34.4% and 38.1%, respectively.

CONCLUSION: Although the prognosis of AML transformed from Ph-neg MPNs is dismal in general, the study suggested a promising result that there were curable patients with allo-HSCT.

Figure 1 Figure 1.

Figure 2 Figure 2.

Disclosures

No relevant conflicts of interest to declare.
The Association of KIR2DS4 and Its Variant KIR1D with CMV Infection after HLA-Matched Sibling Hematopoietic Stem Cell Transplantation
Authors: Yao Yao , Li Xiaoli , Xu Liangjing , Liu Bin , Han Yue , Tian Xiaopeng , Fu Chengcheng , Bao Xiaojing , He Jun , Wu Xiaojin , Depei Wu
Published: November 2019

Objective To expolore the association of KIR2DS4 and its variant KIR1D with cytomegalovirus(CMV) infection after HLA-matched sibling hematopoietic stem cell transplantation.

Methods Polymerase chain reaction with sequence-speciﬁc primers (PCR-SSP) method was used to genotype KIR genes in 267 donor-recipient pairs from Oct 2005 to Apr 2014. Posttransplant monitoring for CMV infection was performed by immune histochemically assays .165 donor-recipient pairs who belong to KIR gene haplotype AA were analyzed for the presence of KIR2DS4 and its variant KIR1D and then further subdivided into the following groups: 2DS4-/1D+ (homozygous for the deletion variant KIR1D), 2DS4+/1D+ (heterozygous), 2DS4+/1D- (two intact KIR2DS4 alleles). Furthermore, we investigated the influence of the KIR2DS4 variants on CMV infection of 165 patients receiving Sibling related HLA matched transplantation.

Results There were no significant differences in frequency of KIR2DS4 or KIR1D between donors and recipients in the haplotype AA group. The ratio of 2DS4+ and KIR1D in haplotype AA group was 2:1.There was no difference on neutrophil engraftment and platelet recovery among the three groups after hematopoietic stem cell transplantation. The CMV infection rate was significantly higher in 2DS4+/1D- group compared with 2DS4+/1D+ group (44.0% vs 19.0%，P=0.002)．In 2DS4-/1D+ group ，the CMV infection rate was higer than that in 2DS4+KIR1D+ group (50.0% vs 19%，P=0.028). However，there was no difference in CMV infection rate between 2DS4+/1D-group and 2DS4-/1D+ group.

Conclusion KIR2DS4 and its variant KIR1D are associated with CMV infection after HLA-matched sibling hematopoietic stem cell transplantation

Disclosures

No relevant conflicts of interest to declare.
Haploidentical Hematopoietic Stem Cell Transplantation (Haplo-SCT) with Pre-Transplant Immunosuppression (PTIS) and Post-Transplant Cyclophosphamide (Post-Cy) in Severe Thalassemia: Safe Approach and Excellent Disease Control
Authors: Hongeng Suradej , Pakakasama Samart , Anurathapan Usanarat , Andersson Borje
Published: November 2019

Background

Currently, the thalassemia free survival rate after hematopoietic stem cell transplant (HSCT) is about 80-90% with either matched sibling or unrelated donor. However, the probability to find a suitable door is only 25-50%. Since most of these patients (pts) could not find the potential donor, we would like to investigate the haploidentical donor HSCT (Hapo-SCT) in thalassemia.

Patients and Methods

Between Jan 2013 and August 2014, 12 thalassemia patients (pts) underwent haplo-HSCT. Six subjects were male and 6 were female. The median age was 16 yrs (range; 2-22). Nine of 12 received stem cells from mother and 3 from father. Ten of 12 were high risk class 3. These high risk class 3 pts received hydroxyurea 20 mg/kg/d for at least 3 months prior to HSCT. All pts received 2 courses of pre-transplant immunosuppression (PTIS) consisting of fludarabine (Flu) 40 mg/m2/d together with dexamethasone (dex) 25 mg/m2/d for 5 days. After 2 courses of PTIS, all pts received a reduced-toxicity conditioning regimen consisting of thymoglobulin 1.5 mg/kg/d (d-11 to d-9), Flu 35 mg/m2/d i.v. (-7 to -2) each dose immediately followed by busulfan (Bu) 130 mg/m2once daily i.v. (d-7 to d -4) for pts > 10 years and 0.95-1.2 mg/kg every 6 hr (d-7 to d-4) for pts < 10 yrs. GVHD prophylaxis consisted of cyclophosphamide (Cy) 50 mg/kg/d (d +3 to d+4). Tacrolimus was given for 6 months to 1 yr started together with mycophenolate mofetil on d+5, the latter was quickly tapered after 2 months. T-cell repleted peripheral blood stem cells (PBSC) were given to all pts, targeting a CD34+ dose of 7-10 x 106cells/kg.

Results

Eleven of 12 were engrafted with full donor chimerism (100%) while one pt suffered graft failure. However, this pt received second transplant on day +30 with minimal conditioning regimen and additional PBSC after which she achieved full donor chimerism. The median time to neutrophil engraftment was 18 days (range; 14 -22). Four pts had acute GVHD gr I, 2 grade II and 1 gr III. Only one had limited chronic GVHD. At this time, all 12 pts survive thalassemia-free and have sustained full donor chimerism (100%). Thalassemia free survial and overall survival rates are 100%. The median follow up time is 12 months (range 4-20).

Conclusion

Haploidentical HSCT for high risk thalassemia pts with our novel approach is safe and should be considered as modality to secure thalassemia-free survival with a low risk for graft rejection and treatment-related mortality. In view of our results, we suggest that all thalassemia pts even those with high risk class 3 features should be offred the chance for cure with HSCT.

Disclosures

No relevant conflicts of interest to declare.
Cyclin D1 C.870G>a Polymorphism in Patients with Multiple Myeloma after Allogeneic Stem Cells Transplantation
Authors: Blau Olga , Slonova Ekaterina , Berenstein Rimma , Nogai Axel , Rieger Kathrin , Pezzutto Antonio , Dörken Bernd , Blau Igor Wolfgang
Published: November 2019

Introduction: Over the last decade, the cyclin D1 (CCND1) c.870G>A polymorphism has been variously reported to confer risk for a large number of cancers. Deregulation of a D-group cyclin is a key feature of multiple myeloma (MM). Previously published meta-analysis shown a relationship between CCND1 c.870G>A and risk of t(11;14) in MM (Wienhold, 2013). Allogeneic stem cell transplantation (AlloSCT) is a potential curative treatment for MM patients. There are no data with regard to the role of CCND1 c.870G>A polymorphism in stem cells donors and outcome of transplantation in MM patients.

Methods: In this study we developed an endonuclease restriction method to identify CCND1 c.870G>A polymorphism. Directly Sanger sequencing were used to confirm the results.

Results: At first, we analyzed 130 MM patients and 100 healthy donors as a control. CCND1 c.870G>A polymorphism was strongly associated with the t(11;14)(q13;q32) (P<0.001). In addition, we have analyzed CCND1 c.870G>A polymorphism 55 pairs of MM patients and their allogeneic stem cell donors, retrospectively. Interestingly, c.870G-genotype in related and unrelated donors was statistically correlated with poor outcome after AlloSCT (P<0.01). Moreover, only patients transplanted with “G-Genotype” donors developed secondary cancer after AlloSCT (P<0.01). No secondary tumors were diagnosed in the group of MM patients received grafts from “A-, AG-genotype” donors.

Conclusion: Our data suggest the published data that constitutive genetic factor (CCND1 c.870 G>A polymorphism) is associated with a specific chromosomal translocation in patients with MM. Despite the fact that our group of patients after AlloSCT is small, we found a statistical association between the c.870G-genotype in donor and a poor prognosis after transplantation.

Disclosures

No relevant conflicts of interest to declare.
Updated Outcomes of NF-08-TM Protocol in HSCT for Patients with b-Thalassemia Major: A Large Prospective Study from Single Center
Authors: Li Chunfu , Wu Xuedong , He Yuelin , Feng Xiaoqin , Liao Jianyun , Zhou Xiaohui
Published: November 2019

Background: The optimization of both erythrocyte transfusion and iron chelating has resulted in a remarkable improvement in the life expectancy of patients with thalassemia major (TM). However, only curative therapy remains allogeneic hematopoietic stem cell transplantation (HSCT). HLA matched sibling transplant (MST) has been commonly used for TM patients with well results. However, this option is unavailable to many patients as a result of a lack of compatible MS, especially in China. Matched unrelated-donor transplant (MUT) and Haploidentical-donor transplant (HIT) have be well performed in malignant disease. But in few thalassemia patients because of high risk of transplantation so far. To expand donor pool and to lower risk of HSCT from alterative donor, we designed a NF-08-TM protocol for MST, MUT and HIT for TM patients since 2009. Outcomes before June 2011 have published in “Blood” in 2012. Here we updated these outcomes.

Aims: to check stability and reliability of NF-08-TM protocol in MST, MUT and HIT for thalassemia patients.

Methods: 293 consecutive patients with TM underwent HSCT between January, 2009 and December, 2013 in our center, including 143 in MUT, 21 in HIT (≥ one HLA mismatch) and 105 in MST. Rate of male to female is 185:108. The median age at transplant was 6 years (rang:0.6-16), The median follow-up time is 29 months (range: 6-64). All patients received the NF-08-TM protocol, which included a new risk classification to adjust dose of IV Busulfex (Bu) and Cyclophosphamide (Cy), and a conditioning regimen of Bu following Cy instead of Cy following Bu. Simultaneously, a intensify graft versus host disease (GVHD) prophylaxis consisted of ATG, Cs A, MMF and short MTX was given.

Results: The estimated 5-year overall survival and TM-free survival were 92.6%, 89.5% and 94.1%, and 90.5%, 89.5% and 92.2% in the MUT, HIT and MST, respectively. The cumulative incidence of graft rejection was 2.1% in total. The cumulative transplant-related mortality was 7.4%, 10.5% and 5.9%, respectively, in the MUT, HIT and MST. Incidence of cytopenia post-transplant (white cell count less than 3.0x109/L for 4 weeks or longer but CMV infection) was 20.6%.

Conclusion: Our large sample prospective study provides results comparing MUT or HIT with the MST for TM patients, which showed that MUT or HIT was comparable with MST when using NF-08-TM-HSCT protocol.

Disclosures

No relevant conflicts of interest to declare.
Clinical Impact of CD25 Expression of Leukemic Blasts on Subsequent Transplant Outcomes in Patients with Refractory or Relapsed Acute Myeloid Leukemia
Authors: Ikegawa Shuntaro , Doki Noriko , Yamamoto Keita , Hino Yutaro , Harada Kaito , Shingai Naoki , Watanabe Ken , Senoo Yasushi , Igarashi Aiko , Najima Yuho , Kobayashi Takeshi , Kakihana Kazuhiko , Sakamaki Hisashi , Ohashi Kazuteru
Published: November 2019

Introduction: We have recently reported that the evaluation CD25 expression on leukemic blasts at the time of disease onset could be an alternative non-molecular tool for predicting transplant outcomes in patients with cytogenetically intermediate acute myeloid leukemia (AML). We herein focus on patients with refractory or relapsed AML and examine the clinical correlation of the surface CD25 expression measured by flow cytometry at the time of transplantation and subsequent transplant outcomes.

Patients and methods: A total of 60 AML samples with primary induction failure (n = 32), first relapse (n = 23) or second relapse (n = 5) were analyzed by means of flow cytometric CD25 antigen expression on leukemic blasts at the time of transplantation. Survival outcomes were compared with regard to the CD25 expression. Overall survival (OS) and progression-free survival (PFS) were estimated by Kaplan-Meiyer. Factors associated with at least borderline significance (p< 0.10) on univariate analyses were subjected to multivariate analysis using backward stepwise proportional hazard modeling. Multivariate analysis was performed using the Cox proportional hazards regression model.

Results: CD25 antigen expression (>10%) on leukemic blasts was observed in 24 patients (40%). There was no significant difference between CD25 positive and negative groups in terms of clinical characteristics including cytogenetic risk, WHO classification, marrow or peripheral blasts % at transplantation, comorbidity, conditioning regimen, donor source or interval between diagnosis and transplantation. Although more early relapse cases within 180 days were observed in the CD25 positive group (p = 0.01), cumulative incidence of relapse, non-relapse mortality, acute or chronic graft-versus-host disease were not different between 2 groups. However, there was a significant difference in OS or PFS between 2 groups: 2-years OS; 8.3% vs. 34.0%, p=0.004 (Fig 1A), 2-years PFS; 8.3% vs. 26.3%, p=0.003 (Fig 1B). Moreover, multivariate analysis showed that CD25 expression was an independent adverse factor for OS (hazard ratio: 2.01, 95% confidence interval: 1.10-3.67, p = 0.02).

Conclusion: Our cohorts with total 60 patients with refractory or relapsed AML showed that CD25 endowed these patients with an adverse prognostic impact, which might not be overcome even by transplantation. AML patients with residual CD25+ blasts at the time of transplantation might require additional therapy before or after transplantation for better survival. However, our small experience clearly desires larger series patients for evaluating the real impact of CD25 expression.

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Race-Based Differences in Routine Cytogenetic Profiles of Patients with Multiple Myeloma
Authors: Blue Brandon Jamaal , Sanfilippo Kristen M. , Ganti Arun , Gumbel Jason , O'Brian Katiuscia , Luo Suhong , Carson Ken R
Published: November 2019

Background: Multiple myeloma (MM) is a common hematologic malignancy for which standard therapy does not offer a cure. Incidence of MM in African-Americans is twice that of Caucasians, suggesting differences in either environmental or genetic risk factors. Historically, black patients have also had a slightly better prognosis than white patients, suggesting racial differences in prognostic factors such as cytogenetics. Since metaphase cytogenetic profiles are routinely collected in MM patients, we sought to determine if race-based differences in cytogenetics exist, using data from the Veterans Health Administration (VHA) cancer registry.

Methods: Using CPT codes, 88271-88275, 88291, 88299, 88365, 83896, 88237, 88261-88264, 88280, 88283, and 88285, we identified 988 patients with MM diagnosed between 1998 and 2009, who also had standard metaphase cytogenetic analysis performed on a bone marrow specimen at the time of diagnosis (228 Black and 585 White) . Fisher’s exact test was used to assess for race-based differences in the following cytogenetic abnormalities: 13q deletion, Hypodiploidy, Hyperdiploidy, and translocations involving chromosome 14.

Results: Among the 988 patients in the cohort, normal cytogenetic profiles, isolated Y chromosome deletion, or no mitotic activity were observed in 704(71%) patients. Translocations involving the immunoglobulin heavy chain (chromosome 14) (n=4) were uncommonly observed on routine cytogenetics, such that no statistical conclusions could be drawn. Hyperdiploidy was noted in 13/228(5.7%) of the black and 45/585(7.7%) of the white patients (p=0.32). Similarly 13q deletions were detected in 8/228(3.5%) black patients, and 21/585(3.6%) of the white patients (p=0.96). Hypodiploidy was also not significantly different in black 4/228(1.8%) and white patients 12/585(2.0%).

Conclusion: This is the largest study of race-based differences in MM cyogenetics presented to date. We found no significant race-based differences in standard metaphase cytogenetics. Future studies should focus on determining if race-based differences can be discovered using fluorescent in situ hybridization (FISH) or molecular testing not available for this retrospective study. Our study adds to this growing body of evidence suggesting that metaphase cytogenetic differences are not a significant factor in MM outcome disparities.

Disclosures

Carson: Spectrum Pharmaceuticals: Honoraria, Research Funding, Speakers Bureau.
Analyzing Survival Trends in Multiple Myeloma Patients in Pre and Post-Bortezomib Era Using the SEER Database
Authors: Manikkam Umakanthan Jayadev , Uprety Dipesh , Kasireddy Vineela
Published: November 2019

Background: Bortezomib is an antineoplastic agent that acts through protease inhibition. Since, its approval in 2003 by U.S. Food and Drug administration, it has been used religiously for multiple myeloma. There is no current population based study that assesses the survival benefit in multiple myeloma from bortezomib. We conducted a population based study to evaluate the relative survival rates in multiple myeloma patients in pre and post-Bortezomib era in the United States.

Methods: We analyzed the Surveillance, Epidemiology, and End Results (SEER) 18 registry database to compare five-year relative survival rates (RS) among multiple myeloma patients. The study arms were categorized by gender (male and female), race (Caucasians and African-Americans) and age (20-59, ≥60 years). The RS were compared between pre-bortezomib era (1991-2002) and post-bortezomib era (2004-2011). The survival rate accompanied standard errors and the statistical significance was defined as p value <0.05.

Results: The database comprised of 57,328 patients. The RS have improved significantly during post-bortezomib era in all cohorts under consideration. There was no significant difference in survival rate between male and female and between the African American and Caucasian. However, the relative survival was much better with the young adult (20-59 years) as compared to adult ≥60 years. The table detailing the relative survival is given below.

Conclusion: The Primary treatment protocols for Multiple Myeloma have changed dramatically since the approval of Bortezomib in 2003. Other novel agents introduced in the last decade include thalidomide, Lenalidomide and liposomal doxorubicin that are used in conjunction with Bortezomib. Survival trends continue to improve and we believe this modest improvement in survival rate is mainly due to the introduction of Bortezomib and other novel agents. The next challenge is to find new drugs which would prevent relapses and further prolong survival. Abstract 2639. Table.Study armSubclass of cohortsPre-bortezomib era (1991-2002)Post- bortezomib era (2004-2011)Z scoreP ValueNRS (%)Relative Standard Error (SE) (%)NRS (%)Relative Standard Error (SE) (%) GenderMale13,06634.60.517,76546.10.615.904<0.0001Female11,58032.10.514,91743.70.614.765<0.0001 RaceCaucasian18,83632.90.423,98145.00.519.259<0.0001African American4,27434.70.86,39145.10.98.488<0.0001 Age group20-596,35848.10.69,43960.20.712.920<0.0001>=6018,28827.80.423,24338.40.516.606<0.0001

Total: male and female: 1991-2002; 24,646, RS 33.4%; SE 0.3%; 2004-2011; 32,682, RS 45.0%, SE 0.4%; Z score 21.766, P value <0.0001.

Disclosures

No relevant conflicts of interest to declare.
Collapse of Ribosome Biogenesis Induces Rapid Erythroblast Differentiation
Authors: Raimbault Anna , Floquet Celia , Guyot Boris , Cuhadar Ulku , Kosmider Olivier , Morle Francois , Mayeux Patrick , Fontenay Michaela
Published: November 2019

Genetic insults of ribosomal protein (RP) genes including heterozygous mutations in Diamond Blackfan Anemia (DBA) or RPS14 haploinsufficiency in acquired 5q- syndrome selectively impair erythropoiesis suggesting that the integrity of ribosome biogenesis (RB) is fully required for the production of 10e11erythrocytes per day in humans. Defective RB consecutive to RPL5, RPL11 mutation or RPS14 deletion is thought to induce cell cycle arrest and a various degree of apoptosis in erythroblasts (EB). However, recent data showed that, loss of RPL5/RPL11neither induces apoptosis nor arrests cell cycle, but hampers the progression through the G1 phase, in primary fibroblasts. Furthermore, RB coordinates cell cycle to cell growth for the maintenance of constant cell size. In this work, we investigate the role of RB in erythroid cell differentiation and cell size regulation.

We first analyzed the renewal of ribosome during in vitro differentiation of human EB derived from cytapheresis CD34+ cells and mouse erythroblasts derived from mouse fetal liver by a pulsed SILAC (Stable Isotopic Labeling by Amino acids in Culture cell) riboproteomic assay. Ribosome biogenesis dramatically decreases with the disappearance of proEB and basophilic EB and the onset of poly- and ortho-chromatophilic EB. Importantly, inhibition of RNA polI by CX-5461 in proEB forced them to enter the final maturation steps with an increase of glycophorin A (GPA) expression. To study the effect of RPS14 heterozygous deletion on RB, UT7/EPO cell line was infected by a lentivirus containing an inducible GFP-shRNA RPS14. After a 48-h treatment with doxycyclin, Rps14 protein expression was reduced by half and sorted GFP-positive cells had an altered ribosome profile devoid of 40S small subunit or 80S entire particle. Consistently, RB inhibition induced a cell size reduction. Second, we compared RB level in cells responsive to SCF+EPO or EPO alone. RB was optimal when EB responded to SCF+EPO and this was correlated with cell size being higher in SCF+EPO-responsive cells compared to EPO-responsive cells. Both cytokines additively activate the cell growth regulator, p70S6K1. Third, inhibition of p70S6K1 by rapamycin, or a specific chemical S6K1 inhibitor significantly reduced RB as shown by a 50% decrease of ribosome renewal in pulsed-SILAC. Inhibition of RB by rapamycin led to a size reduction and to GPA acquisition, which are the features of erythroid cell differentiation.

Our data shows that the collapse of RB due to the loss of c-Kit and reduced activation of p70S6K1 is a key step for cell growth inhibition and induction of terminal differentiation in human or mouse erythroblasts.

Disclosures

No relevant conflicts of interest to declare.
Sphingosine 1-Phosphate Induces Erythrocytes Sickling in Sickle Cell Disease By Promoting Hemoglobin S Aggregation
Authors: Zhang Yujin , Berka Vladimir , Song Anren , Kellems Rodney E , Xia Yang
Published: November 2019

Sickle cell disease (SCD) is a genetic hemolytic disease with high morbidity and mortality affecting millions of individuals worldwide. Although SCD was identified a century ago, we still lack effective mechanism-based therapies to treat this disease. Our recent study showed inhibition of sphingosine kinase 1 (Sphk1, the enzyme that produces S1P) significantly decreased S1P level in sickle erythrocytes, attenuated sickling cells. We also found sickling was induced by high concentration (10µM) but not low concentration (0.5 µM) of S1P, which indicates S1P induced sickling is independent of its receptors. However, the mechanism of S1P inducing sickling in SCD is unknown. Because sickling is induced by formation of polymers of deoxygenated sickle hemoglobin, we hypothesized that S1P functions intracellularly to induce sickling by directly promoting polymerization of deoxygenated HbS. To test this possibility, we determined the effects of S1P on deoxy-HbS polymerization using temperature jump method. We found that 5µM S1P began to induce aggregation of deoxy-HbS (0.25mM) with shorter delay time at 17.65 ± 0.57min compared to HbS treated with solvent (methanol) which with delay time at 19.28 ± 0.17 min. Moroever, S1P at 10µM induced deoxy-HbS aggregation with further decreased delay time at 15.71 ± 0.38 min, clearly demonstrating a dose-dependent effect. Thus, this study indicates that S1P at concentrations higher than 5µM dramatically and specifically accelerated HbS aggregation. Next, we found that aggregation amplitude (ΔA) of deoxy-HbS was significantly increased by S1P at a concentration of 5µM and was further elevated at 10mM of S1P. indicating that S1P enhanced polymerization ability of deoxy-HbS in a dosage-dependent manner. Finally, we detected the effect of S1P on deaggregation of aggregated deoxy-HbS. We found that S1P significantly reduced temperature drop-mediated unaggregated HbS and this effect was in a dosage dependent manner. In order to test whether anti-sickling drug can increase delay time of Hb S aggregation, we chose 5-HMF that was reported anti-sickling. We found 5-HMF significantly increased delay time of HbS. Taken together, we demonstrated that S1P induces HbS polymerization via three mechanisms: 1) Accelerating polymerization of HbS by shortening temperature jump-mediated delay time; 2) enhancing the aggregate formation during temperature jumping; and 3) reducing deaggregation ability during temperature dropping. Thus, this study strongly supports our in vivo animal, in vitro cellular and functional evidence that S1P contributes to sickling by promoting polymerization.

Disclosures

No relevant conflicts of interest to declare.
Reversibility of Iron Loading in Transferrin-Deficient Mice
Authors: Bartnikas Thomas Benedict
Published: November 2019

An abundant serum metal-binding protein, transferrin is a critical factor in mammalian iron homeostasis. As transferrin is essential for iron delivery for erythropoiesis, transferrin-deficient patients and mice develop anemia but also a paradoxical systemic iron overload. Iron overload results from deficiency in hepcidin, a peptide hormone synthesized largely by the liver that inhibits dietary iron absorption and macrophage iron efflux. Using transferrin-deficient mice, we have previously shown that transferrin deficiency leads to hepcidin deficiency via two mechanisms. First, the anemia and/or hypoxia that results from inadequate iron delivery to bone marrow actively suppresses hepcidin expression by the liver. Second, transferrin directly stimulates hepcidin expression by the liver independently of transferrin's role in erythropoiesis; this second mechanism is dependent upon hemojuvelin, a bone morphogenetic protein coreceptor essential for hepcidin expression that is mutated in juvenile hemochromatosis. Here we present data examining the reversibility of iron loading in transferrin deficiency. Measurement of tissue metal levels in adult transferrin-deficient mice demonstrated progressive and systemic iron loading yet minimal to moderate changes in levels of other metals. Treatment of adult transferrin-deficient mice with exogenous transferrin normalized red cell parameters and hepcidin levels and reduced iron load in most organs. To further investigate the reduced tissue iron burden in transferrin-treated mice, we employed iron-loaded cell lines to demonstrate that cellular iron efflux can be stimulated by transferrin and that this efflux is specific to iron but not manganese or zinc. Current studies focus on two goals. First, we are determining if the reduction in tissue iron load represents solely a redistribution of iron stores to the erythron or additionally excretion of iron via urine and/or feces by measuring whole body iron levels in mice before and after transferrin treatment. Second, we are identifying transporters required for mobilization of iron stores using expression profiling of known and putative iron transporters in mouse tissues and validating the role of transporters of interest using our cell culture model. Results of these studies and their implications will be presented.

Disclosures

No relevant conflicts of interest to declare.