bluuebunny/crossref_metadata_2025 huggingface dataset:
Implementation of a Hematological Malignancy-Based Survivorship Clinic.
Authors: Steinberg Amir , Asher Arash
Published: October 2019

Abstract 4550

BACKGROUND

There are approximately twelve million cancer survivors in the United States. Cancer survivors are at increased risk for health complications such as heart disease, secondary malignancies, and psychological distress. Reflecting the significant needs of cancer survivors as part of our health care system, the Institute of Medicine issued a seminal report, From Cancer Patient to Cancer Survivor: Lost in Transition, recognizing cancer survivorship as a “distinct phase of cancer care.” A dedicated survivorship clinic with creation of personal care plans and treatment summaries can serve the needs of this growing cohort of patients.

OBJECTIVE

To determine the viability of a hematological malignancy-based survivorship clinic with treatment summary and care plan for survivors. The malignancies include all lymphomas, leukemias, plasma cell dyscrasias, myeloproliferative disorders, and hematopoietic stem cell transplant patients.

METHODS

A literature search was initiated examining recommendations and guidelines regarding follow-up of cancer patients. We incorporated and consolidated guidelines from ASCO, NMDP, NCCN, and COG, pertinent articles from the MEDLINE/PubMed database, and the growing literature from cancer survivorship textbooks and lectures. Awareness of the program was to be initiated with letters to affiliated physicians, lecture series, patient-oriented educational handouts, and articles in the hospital literature.

DATA SYNTHESIS

A treatment care plan was devised for all survivorship clinic patients. This care plan summarized patients' prior care with recommendations for follow-up – such as cardiac and cancer screening, osteoporosis, thyroid disease, and vaccination schedules–all based on our consolidative review of available evidence-based or expert guidelines. A referral base was developed with specialists focused on lymphedema, psychiatric issues (needs determined by screening tools), social work, art therapy, exercise, and nutrition.

RESULTS

Patients from affiliated hematologists/oncologists were seen in the survivorship clinic. Unique care plans were devised for them. A satisfaction survey was to be implemented. Preliminary satisfaction from both physicians and the patients themselves was positive.

CONCLUSION

A hematological-based survivorship clinic is feasible and fills an important need in our large, urban, hospital-based cancer center. Further investigation will focus on developing a computer-based treatment summary and care plan, increasing patient referrals, and developing a lecture series devoted to survivorship issues.

Disclosures:

No relevant conflicts of interest to declare.
Effectiveness of a Novel Humanized VB22B Minibody against Human TPO Receptor Compared with Recombinant TPO and Eltrombopag Using Primary Human CD34+ Cells and Platelets.
Authors: Matsuki Eri , Yamane Akiko , Okamoto Shinichiro , Miyakawa Yoshitaka
Published: October 2019

Abstract 4559

Thrombopoietin (TPO) is a cytokine produced primarily by the liver and kidney that regulates platelet production by stimulating proliferation and differentiation of hematopoietic stem cells, megakaryocytic progenitor cells and megakaryocytes via activation of its receptor, c-Mpl. Recently, TPO receptor agonists such as eltrombopag and romiplostim have been approved for chronic ITP. huVB22B was created as a novel humanized form of murine sc(Fv) 2VB22B minibody (BLOOD, 2005) which activates human c-Mpl by CDR grafting. The advent of these various TPO receptor agonists prompted us to consider the differences in their mechanisms of action, efficacy or potency. However, to date, there has been no in vivo or in vitro study directly comparing the effects of different TPO receptor agonists. In this study, we compared the efficacy of huVB22B on CFU-GM, CFU-E, CFU-Megakaryocyte (CFU-MK), megakaryocyte maturation (DNA ploidy and proplatelet formation) with those of recombinant human TPO (rhTPO) and eltrombopag. Primary human CD34+ bone marrow cells were cultured with various concentrations of rhTPO, huVB22B and eltrombopag using methylcellulose based media. In serum-free condition, 0.286 nM rhTPO, 0.182 nM huVB22B and 17.7 mcM eltrombopag demonstrated almost equivalent efficacy of megakaryocyte colony formation. At these concentrations, all agents demonstrated similar in vitro efficacy for colony formation of CFU-GM and CFU-E, proplatelet formation and nuclear maturation of megakaryocytes. In preliminary results, huVB22B induced maturation of CFU-MK earlier than rhTPO and eltrombopag, suggesting that huVB22B might have some potential to increase human platelets faster than other agents in vivo. This is compatible with the observation that huVB22B induced tyrosine phosphorylation of STAT3, STAT5 and JAK2 faster and stronger than rhTPO and eltrombopag in human primary platelets. Both rhTPO and huVB22B enhanced low-dose ADP and collagen-induced human platelet aggregation in vitro. In contrast, eltrombopag did not enhance ADP or collagen-induced platelet aggregation, although it induced activation of JAK-STAT pathway in human platelets. Contrary to the fact that huVB22B induces phosphorylation of intracellular signaling molecules faster and stronger than rhTPO in human platelets, the priming effect by huVB22B on platelet aggregation was much weaker than rhTPO. In conclusion, we confirmed that newly created huVB22B minibody induced colony formation of CFU-MK, CFU-E, CFU-GM and maturation of megakaryocytes from human bone marrow-derived CD34+ cells in vitro. The differences among TPO receptor agonists observed in our study would lead to further understanding of the basic biology of megakaryopoiesis and the action of TPO receptor agonists.

Disclosures:

Okamoto: Alexion: Research Funding. Miyakawa:GlaxoSmithKline: Consultancy.
CXCR4 Blockade as a New Targeted Therapy for Acute Myeloide Leukemia Characterised by High Cell Surface Density of CXCR4.
Authors: Zhang Yanyan , Jouni Dima , Wittner Monika , Bouamar Hakim , Dombret Herve , Coppo Paul , Opolon Paule , Fujii Nobutaka , Ito Mamoru , Vainchenker William , de Botton Stephane , Louache Fawzia
Published: October 2019

Abstract 4570

Stromal cell-derived factor 1 (SDF-1)/CXCR4 axis plays key roles in hematopoiesis regulating the interactions between hematopoietic cells and their stromal microenvironment within the bone marrow, their trafficking from the bone marrow to blood, their proliferation and survival. SDF-1/CXCR4 interactions also participate in the development of leukemic blasts in acute myeloid leukaemia (AML) influencing their trafficking, survival and differentiation. We used the xenotransplantation model of nonobese diabetes/severe combined immunodeficiency γcnull (NOG) mice reconstituted with human primary cells from AML patients and 2 small molecule competitive antagonists of CXCR4, AMD3100 and TN140 to better define the role of CXCR4/SDF-1 on the leukemic blast burden within the bone marrow and in extramedullary sites. In a first set of experiments, we investigated whether there was a correlation between CXCR4 expression or function on leukemic blasts and their ability to engraft the bone marrow of NOG mice in 34 patients. Using flow cytometric analyses, we observed that CXCR4 membrane expression was highly variable between patients. This expression did not correlate with engraftment. In addition, SDF-1 responsiveness evaluated by transwell migration only marginally correlated with engraftment. However, in these initial analyses, we observed that the differences between engrafters and nonengrafters were significant if the cut-point migration is set to 20%. Patients with higher chemotactic response to SDF-1 had a significantly increased NOG repopulating ability. In further studies, NOG mice reconstituted with AML cells from 5 different patients were treated with optimal concentration of either AMD3100 or TN140 for 1 week. We observed that CXCR4 inhibition by TN140 (used as a single therapy) had profound inhibitory effects on the proliferation and the development of extramedullary dissemination of the disease in this xenotransplantation. This patient had FAB M1 leukemia and initially exhibits high CXCR4 level on blast. Our study demonstrated that CXCR4 inhibition in selected patients might be a potent therapy against leukemic development.

Disclosures:

Bouamar: Association pour la Recherche sur le Cancer: Employment; Cancéropôle IDF: Employment.
Human Umbilical Cord Wharton's Jelly-Derived Mesenchymal Stem Cells Differentiate Into Insulin-Producing Cells.
Authors: Ma Lian , Wang Hongwu , He Hongyan , Lin Limin , Li Weizhong , Huang Tianhua , Ma Guixia , Wang Aihong
Published: October 2019

Abstract 4578

Introduction

Islet transplantation is an effective way of reversing type 1 diabetes. However, islet transplantation has been hampered by problems, such as immune rejection, and the scarcity of donor islets. Human Umbilical Cord Wharton's Jelly-derived Mesenchymal Stem Cells (huMSCs), which can be differentiated into insulin-producing cells could provide a source of cells for transplant.

Methods

Vitro Research We isolated and cultured huMSCs, and induced huMSCs differentiated into insulin-producing cells in the condition of islet cells grows. The morphology of huMSCs after induction were monitored by under inversion phase contrast microscope?GImmunocytochemical methods were used to detect the insulin and glucagon protein, and reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect Human insulin gene and PDX-1 gene. Dithizon-stained was used to detect zinc hydronium and radio-immunity was used to detect insulin level of culture supernatant.Vivo Research huMSCs were transplanted into the body of diabetic rats through vena caudalis, and then we observed the change of blood glucose?Abody weight ?Aserum insulin levels and survival ratio in STZ-induced diabetic rats. We detected human insulin by immunohistochemistry and RT-PCR. HE stain was used to detect the morphological changes of rat's pancreatic island.

Results

Vitro Research The morphology of huMSCs under medicine induction gradually changed from fibroblast to round and some of then had the tend of forming clusters.?GThe result of immunocytochemical showed that the expression of human insulin and glucagon was positive after treatment with medicine?GhuMSCs induced by medicine can express insulin and PDX-1 gene by RT-PCR?GDithizon stain show that the cytoplasm of huMSCs after induction were stained in Brownish red color?Gthe results of radio-immunity manifested that the insulin quantity secreted by medicine induction were significant differences compared with control group(t??6.183,P<0.05). Vivo Research When transplanted into Streptozotocin(STZ)-treated diabetics rats, huMSCs can decreased blood glucose, increased body weight and survival ratio in diabetic rats?GAfter being transplanted for one month, we discovered that it can be planted into rat's pancreas and liver by Hoechst33258?Gimmunohistochemistry and RT-PCR show that the pancreas of rat can express human insulin?Gthe morphology of rats' pancreatic island was repaired obviously if compared with diabetic rats before the transplantation through HE-stain.

Conclusion

huMSCs can be differentiated into insulin-producing cells in vitro or in vivo. Therefore, huMSCs have the potential to become an excellent candidate in β cell replacement therapy of diabetes.

Disclosures:

No relevant conflicts of interest to declare.
Differentiation of Human Male Germ Cells From Human Umbilical Cord-Derived Mesenchymal Stem Cells in Vitro and In Vivo.
Authors: Ma Lian , Wu Xiaoying , Lin Limin , Lin Guangyu , Tang Qiuling , Huang Tianhua , Feng Xueyong , Ma Guixia , Xie Qingdong
Published: October 2019

Abstract 4581

Introduction

Infertility affects 15% of couples, about 50% infertility caused by male and growing evidence suggested an increasing problems in male reproductive. Recent studies have demostrated that adult stem cells have more flexible potentials than expected, and possessed the plasticity and capacity to transdifferentiate into mutilineage cells, including germ cells. Human umbilical cord-derived mesenchymal stem cells (HUCMSCs) possess stem cell properties. In this study, we cultured HUCMSCs, and assessed the possibility of HUCMSCs differentiated into human male germ cells in vivo and in vitro, and find a new source of cells for the transplantation to the male infertility.

Methods

The ethics committee of our institution approved this study. HUCMSCs were isolated from the Wharton's jelly of the umbilical cord, clonally expanded. To investigate the capacity of differentiation in vitro, HUCMSCs were treated with human menopausal gonadotropinn (HMG) and retinoid acid (RA) in vitro. While investigate the capacity of differentiation in vivo, HUCMSCs were transplanted into the seminiferous tubules of busulfan-treated mice testes after labeled with pIRES2-EGFP or bromodeoxyuridine (BrdU). After induction in vitro, the morphologic changes of the differentiated cells were detected by phase contrast microscopy?Aelectron microscopy and transmission electron microscope?Gthe male germ cell markers were detected by immunohistochemistry, Western-blot and PT-PCR. HUCMSCs were also transplanted into the seminiferous tubules of the busulfan-treated mice by microinjection. To assess the fate of HUCMSCs in the testis, the survival?Amigration and germ cell markers of the HUCMSCs in the infertility mice testis were detected by immunohistochemistry?A immunofluorescence.

Results

HUCMSCs can express some some molecular markers of germ cells after induction. Immunohistochemistry revealed that HUCMSCs can survive in the mice testis at least 120 days, and they can migrate from the lumens to the basement membrane then to lumens again. Immunofluorescence showed that HUCMSCs can go further differentiation in the mice favorable testicular environment, and express the germ cell markers.

Conclusions

These suggested that HUCMSCs may differentiate into male germ cell-like cells after induced by HMG and RA in vitro; and it could survive also in a favorable testicular environment, may differentiate into germ cell lineages. This finding may provide a new strategy for the treatment of male infertility.

Disclosures:

No relevant conflicts of interest to declare.
Distinct Mechanisms Regulate the Expression of flt3 Gene in Normal and Leukaemia-Like Stem Cells.
Authors: Volpe Giacomo , Walton David , Dassé Emilie , del Pozzo Walter , O'Neill Laura , Göttgens Berthold , Frampton Jonathan , Dumon Stephanie
Published: October 2019

Abstract 4586

The interaction between the tyrosine kinase receptor Flt3 and its ligand FL leads to crucial signalling during the early stages of the commitment of haematopoietic stem cells (HSCs). Consequent to mutations or over-expression, constitutive activation of the Flt3/FL pathway is also a key factor in the enhanced survival and expansion of leukaemic cells. Flt3 expression is in particular a feature of Acute Myeloid Leukemia (AML) as it is found in the vast majority of cases. As well as a powerful inducer of leukaemogenesis, Flt3 expression could therefore be viewed as a measurable consequence of the leukaemic transcriptional programme. Hence, understanding its transcriptional regulation would improve our knowledge of the molecular events underlying normal stem cell commitment and AML disease. Accounting for 40% of AML cases, normal cytogenetic AML (NC-AML) is a heterogeneous group of diseases that present a number of different alterations in transcriptional regulators of haematopoiesis, including mutations (MLL, C/EBPα, AML1, PU.1, GATA-1) or altered expression brought about by silencing (C/EBPα, PU.1) or activation (MN1, MLL, ERG1, Cdx2, Hox proteins). Here, we show that a substantial number of these transcription factors are implicated in the regulation of the flt3 gene, and by comparing cell line models of normal HSCs and AML, we pinpoint the importance of HoxA9, Meis1, C/EBPα and c-Myb, and remodelling of the epigenetic environment in the regulation of flt3 expression in leukaemic cells. C/EBPa mutations are found in 10% of AML cases and are considered to be associated with a good prognosis in the absence of Flt3 constitutive activation. We present a model linking C/EBPα and c-Myb activity to flt3 gene expression and FLT3 signalling.

Disclosures:

No relevant conflicts of interest to declare.
Follicular Dendritic Cell (FDC)-Induced Microrna-Mediated up-Regulation of PRDM1 and Down-Regulation of BCL6 in Germinal Center B Lymphocytes: A Potential Mechanism for B-Cell Differentiation.
Authors: Lin Jianhong , Lwin Tint , Zhao Jianjun , Zhao Jie , Crespo Luis , Sotomayor Eduardo M. , Moscinski Lynn C. , Dalton William S. , Wright Ken , Tao Jianguo
Published: October 2019

Abstract 4588

B-cell differentiation process is tightly regulated by suppression or induction of specific transcription factors. Among various transcriptional regulators, BCL6 and PRDM-1 are master regulators for germinal center (GC) formation and terminal B-cell differentiation. Dysregulation of BCL6 and PRDM-1 have been associated with lymphomagenesis. However how these transcription factors are regulated and what determines their expression are unclear. Given that follicular dendritic cells (FDC) closely interact with B cells within the GC, provide survival signal to protect B cells from apoptosis and are essential for the differentiation of GC B cells, we used an in vitro FDC-B-cell co-culture model to explore the role of FDC-B cell interaction and FDC-induced miRNA in the regulation of BCL6 and PRDM-1 expression. In this study 1) we revealed that follicular dendritic cells (FDCs, HK) regulate expression of transcription factor (BCL6, and PRDM1) via cell-cell contact, 2) we showed that FDCs regulate expression of B-cell survival and differentiation-related microRNAs, 3) we demonstrated that microRNAs regulate expression of transcription factors BCl6 and PRDM1 and 4) we documented that follicular dendritic cells regulate expression of transcription factor (BCL6, and PRDM1) through microRNAs and plays an important role in B-differentiation. These studies establish new molecular mechanisms for regulation of BCL6 and PRDM-1. FDC-induce miRNA mediated down- and up-regulation of transcriptional factors may contribute to the phenotype maintenance of GC, and pathogenesis of non-Hodgkin's lymphoma (NHL) by interfering with normal B-cell terminal differentiation.

Disclosures:

No relevant conflicts of interest to declare.
In Vitro Knock-Down of Myh9 Leads to a Myh9-Related Disorder Phenotype in Human Megakaryocytes.
Authors: Chen Yolande , Boukour Siham , Romdhane Monia , Seye Ababacar , Bluteau Olivier , Lordier Larissa , Chang Yunhua , Galy Anne , Favier Rémi , Nurden Paquita , Raslova Hana , Vainchenker William , Debili Najet
Published: October 2019

Abstract 4596

Normal platelet production is dependent on the formation of branched long cytoplasmic extensions, called proplatelets (PPT). Mutations of the Myh9 gene (encoding for the nonmuscle myosin heavy chain IIA) result in autosomal dominant disorders, where patients develop various degrees of macrothrombocytopenia, with sometimes glomerular impairment, hearing loss and cataracts. There has been questioning as to whether the mechanism for the macrothrombocytopenia is haploinsufficiency, or a dominant negative effect of the mutated gene. We performed an in vitro study to investigate PPF from patient megakaryocytes (MK). By this approach, a decrease in PPF from patient CD34 derived MKs was observed in comparison to normal cultured MK. Surprisingly this defect of PPF observed in patients was rescued by blebbistatin, an inhibitor of class II myosin. Immunofluorescence studies performed showed that besides clusterization of GPIb in patient's platelets, no major repartition abnormalities were seen in cultured MKs derived from patient's CD34 for myosin, actin, tubulin, vWF, and Rac (except in one patient where actin and Rac formed aggregates to some extend, in a small number of MKs). In order to better understand the role of myosin during normal and abnormal PPF, we used a shRNA strategy to disrupt the Myh9 expression during normal MK differentiation and compared shRNA-treated MKs with MKs derived from patient CD34. Megakaryocytes treated with a shRNA that knocks down the protein of about 50%, did not alter MK ploidization, but decreased in vitro PPF, as previously observed for cells issued from patients. Moreover, shRNA-treated MKs exhibited the same ultrastructural abnormalities as patient MKs. Addition of Blebbistatin to shRNA treated MKs led to an increase of PPF, suggesting that the remaining myosin II might be hyperactivated and inhibit PPF.

Altogether this study strongly suggests that the thrombocytopenia of the Myh9 syndrome is essentially related to haploinsufficiency in myosin II.

Disclosures:

No relevant conflicts of interest to declare.
Splenomegaly with Increased Erythroblast Numbers During Erythropoietic Stress Caused by Ambient Low-Temperature in Mice.
Authors: Maekawa Shun , Iemura Hitomi , Miyazaki Hiroshi , Kato Takashi
Published: October 2019

Abstract 4602

Erythropoietic stress such as hypoxia has been well described in mammals. These conditions decrease oxygen supply and then enhance production of erythropoietin (EPO) regulates production of red blood cells. Moreover, spleen becomes the main organ of erythropoiesis due to limited marrow space. Here, we describe a new erythropoietic stress, ambient low-temperature. It has been reported that peripheral blood cell counts are affected by ambient low-temperature in several vertebrates. Cold-acclimated rat and chicken exhibit polycythemia (Dveci et al, J Comp Physiol, 2001; Yahav S, Poult Sci, 1997). The response is considered to increase demand of tissues for oxygen, and then enhance metabolic rate and capacity for heat production to acclimate to ambient low-temperature. However, the physiological mechanisms had not been investigated. First, we examined peripheral erythrocyte levels in C57BL/6 mice putting into 5°C ambient. Hematocrits increased from 48% to a plateau of 53% after fourteen days. Likewise, hemoglobin concentration, initially 15 g/dl, rose to 17 g/dl. Reticulocyte production index significantly increased from 4% to 8% after seven days. These data suggested that mice exposed to low-temperature enhanced production of erythrocytes, so we next examined the anatomy and cell composition of their spleens. On day 5, spleens were about 6 mg/g of body weight, two-fold those on day 0. They gradually decreased to their initial weights on day 14. Flow cytometry showed 38% more Ter119+ splenocytes and four-fold more CD71 high Ter119+ early erythroblasts than normal. These values also gradually declined to their initial numbers by day 14. The results suggested the elevated red blood cell counts were due to an increase in production. To test erythropoietic activity in serum, we used an erythrocyte colony-forming assay. Serum from mice kept at low-temperature showed no ability to stimulate CFU-E colony formation in vitro. However, a combination of the serum with EPO (0.5 U/ml) increased CFU-E numbers 1.5 to 2 times that of a combination of normal serum plus EPO or EPO alone. Whether inducible factors account for these effects is the focus of future investigation. Our findings suggest a low-temperature environment is an erythropoietic stress that may offer insights into the adaptive physiology of red blood cell production in mice.

Disclosures:

No relevant conflicts of interest to declare.
Pain Rate and Social Circumstances Rather Than Cumulative Organ Damage Determine the Quality of Life in Adults with Sickle Cell Disease.
Authors: van Tuijn Charlotte F.J. , van Beers Eduard J. , Schnog John-John , Biemond Bart J.
Published: October 2019

Abstract 4605

Background

Previous studies have demonstrated a significantly reduced quality of life (QoL) in sickle cell patients which appeared to be related to the significant morbidity associated with sickle cell disease such as frequent painful crises. With the longevity of sickle cell patients in the Western world chronic organ damage may also be an important factor to the QoL of sickle cell patients. Furthermore, not directly disease related factors such as socioeconomic circumstances may also be important in the general well being of these patients.

Aim of the study

First, to determine the contribution of chronic organ damage and cumulative sickle cell related complications to QoL of sickle cell patients in the Netherlands. Second, we analysed the effect of socio-economic circumstances on QoL in our patients.

Methods

Consecutive adult sickle cell patients cared for in a teaching hospital were included. QoL was assessed with SF-36 forms and was represented in a physical component scale (PCS) and a mental component scale (MCS). In all patients a systematic evaluation of chronic organ damage was performed including: pulmonary hypertension, renal failure, microalbuminuria, retinopathy and iron overload. In addition, sickle cell related complications including: the frequency of painful crises, chronic leg ulcers, avasculair osteonecrosis, stroke, acute chest syndrome and priapism were assessed in the previous five years by chart review. We studied the QoL scores in relation to presence of chronic organ damage, sickle cell related complications and socioeconomic circumstances (patients occupation and educational level).

Results

A total of 96 patients were included in our study. The presence of chronic organ damage was not related to the QoL of our patients despite the incidence of pulmonary hypertension in 24% of our patients. Of the sickle cell related complications in the previous five years, pain rate was significantly associated with mental component score (MCS) but not with the physical component score (PCS). With respect to the socioeconomic circumstances, 35% of the sickle cell patients were unemployed as compared to 6% of the general population and 16% of immigrants without SCD. Both occupation and the level of education were significantly related to PCS while no relation with MCS or pain rate was found.

Conclusion

In sickle cell patients in Amsterdam, the Netherlands, QoL is mainly determined by pain rate and socioeconomic circumstances such as occupation and educational level. Chronic organ damage, although a major factor determining life expectancy in SCD, was not a determinant of QoL.

Disclosures:

No relevant conflicts of interest to declare.
Current Epidemiology and Hospitalization Characteristics for Acute Chest Syndrome: A Nationally Representative Survey.
Authors: Goel Ruchika , Viswanathan Prabhu , Krishnamurti Lakshmanan
Published: October 2019

Abstract 4608

Background

Acute chest syndrome (ACS) is a major cause of morbidity and the leading cause of mortality in patients with sickle cell disease (SCD). While the causes, outcome, and response to therapy in ACS has been described before, there is scant literature on current nationally representative estimates of burden, characteristics and cost of hospitalizations for SCD patients with ACS.

Methods

The Nationwide Inpatient Sample (NIS) database, sponsored by the Agency for Health Care Quality and Research, is a stratified probability sample of 20% of all hospital discharges among U.S. community hospitals (n=1,044, sampling universe = 90% of all such discharges). Sampling weights were applied to represent all community hospital discharges in the US for the year 2007.

Presence of following ICD-9-CM diagnosis codes were used to define SCD related hospitalizations: 28241, 28242, 2825, 28260, 28261, 28262, 28263, 28264, 28268, and 28269. ICD-9-CM 517.3 was used to identify ACS as a diagnosis during hospitalization for SCD. Only data on SCD related hospitalizations were analyzed.

Results

In 2007, there were 86,318 discharges nationwide with a primary diagnosis and 79,766 with a secondary diagnosis of SCD (total=166,084). Of these, 7039 (4.2%) had ACS as a complication. Though, males represented 37.5% of SCD related hospitalizations but accounted for 52.1% of ACS episodes. ACS was more common complication in those with age <18 yrs than 18-44 age group (8.1% vs. 3.5%)

Pulmonary hypertension as one of the discharge diagnosis was reported in 4.7% of the patients with ACS as compared to 2.2% of the patients without ACS (p <0.001). At least one simple transfusion was seen in 43.0% of SCD patients admitted with a ACS compared to 21.7% without ACS (p-value <0.001). Exchange transfusion (3.6% vs. 0.4%; p <0.001) and mechanical ventilation (2.2% vs. 0.7%; p<0.001) were more frequently among hospitalizations with ACS than without it.. The mean (SD) length of hospitalization in SCD patients with ACS was higher (6.9±0.3 vs. 5.2±0.1; p<0.001) than those without ACS. The mean length of stay by various age categories is depicted in Table 1. Presence of ACS was associated with significantly higher mean charges per hospitalization (USD 35145 vs. USD 22836; p<0.001) Higher in-hospital mortality (1.3% vs. 0.5%, p<0.001) was seen among SCD hospitalizations with ACS as complications than without. The majority of deaths were in the 18-44 years age group (69.9%) followed by 18.2% deaths in <18 years group.

Conclusions

ACS is a major and common complication in hospitalizations with SCD as a discharge diagnosis. Among SCD patients, presence of ACS is associated with an increased risk of receiving a simple or exchange transfusion, mechanical ventilation, increased length of hospital stay, higher hospital charges and in-hospital mortality.

Disclosures:

No relevant conflicts of interest to declare.
Improved T Cell Recovery After Transplantation of CD3/CD19 depleted Haploidentical Stem Cell Grafts in Pediatric Patients.
Authors: Teltschik Heiko-Manuel , Feuchtinger Tobias , Pfeiffer Matthias , Schumm Michael , Mueller Ingo , Handgretinger Rupert , Lang Peter J.
Published: October 2019

Abstract 4652

Profound depletion of T and B cells is a fundamental prerequisite for haploidentical transplantation and allows to minimize GvHD despite HLA incompatibility. However, posttransplant recovery of donor derived T cells is delayed after various graft manipulation procedures and may result in severe infections. Methods to improve this recovery are of great importance. Here we present immune reconstitution data in patients who received CD3/CD19 depleted stem cells in combination with melphalan based or standard conditioning regimens.

32 patients with ALL (n=14), AML/MDS (n=17), CML (n=1) were included. T and B cells were directly depleted using antiCD3/antiCD19 coated magnetic microbeads and the CliniMACSTM device.

The patients received either TBI or Bu i.v. and OKT3 (n=9) or a reduced intensity conditioning (“RIC”: Mel 140mg/m2, Flud 160mg/m2, TT 10mg/kg, OKT3, n= 23).

Absolute numbers of lymphocyte subsets per microliter on day 90 were compared within these both groups and with a historical control group (patients with leukemias who received CD34 selected grafts and TBI or Bu based standard conditioning regimen in combination with ATG, n=28).

CD3+4+, CD3+8+ and total numbers of CD3+ of patients after CD3/CD19 depletion were significantly higher in the RIC-group than in the TBI/Bu-group (mean numbers: 85.83 vs. 38.84; 133.46 vs. 19.69; 270.27 vs. 63.99; p<0.05, unpaired t-test).

Comparison with the whole CD34 historical group showed a faster recovery of CD3+4+ in patients with CD3/19 depletion and RIC (104.26 vs. 54.22; p= 0,034) but no significant difference in CD3+8+ and CD3+. Furthermore, subgroups of the CD34 historical population were compared: patients with CD3/19 depletion and RIC had a significantly faster recovery of CD3+4+, CD3+8+ and CD3+ than CD34 patients with TBI (104.26vs. 25.48; 133.46vs. 43.17; 270.27 vs. 65.86; p<0.05) but had no advantage over CD34 patients with non-TBI conditioning.

Conclusions: the type of graft manipulation appeared to have an influence on the speed of CD4+ recovery (CD3/19 depletion > CD34 selection). Moreover, the use of TBI had a clear negative impact on all T cell subsets: patients with TBI had a slower recovery than patients with non-TBI conditioning, independent from graft manipulation procedures and probably due to thymic damage. Thus, the use of RIC-protocols in combination with CD3/CD19 depletion may help to speed up the immune recovery after haploidentical transplantation. Further studies are warranted to evaluate the risk of relapse.

Disclosures:

No relevant conflicts of interest to declare.
HHV-6 Reactivation Lowers the Risk of Acute Graft-Versus-Host Disease but Enhances the Risk of Infections Due to Activation of CD4+FoxP3+ Lymphocytes.
Authors: Lange Andrzej , Jaskula Emilia , Madej Sylwia , Dlubek Dorota
Published: October 2019

Abstract 4653

CMV, EBV, and HHV-6 reactivation was monitored for 20 days to 4 years after allogeneic hematopoietic stem cell transplantation (alloHSCT) with respect to hematological and immunological reconstitution as well as fate in 118 patients (17 children and 101 adults) who received alloHSCT (100 peripheral blood progenitor cell (PBPC) and 18 bone marrow transplants from 53 siblings and 65 unrelated donors, 64 patients receiving reduced intensive conditioning (RIC) and 54 myeloablative conditioning). All patients on RIC and those transplanted from unrelated donors received anti-thymocyte globulin (ATG). Herpes virus DNA copies were measured by quantitative PCR (primers and probes designed according to Kimura 1999, Machida 2000, and Locatelli 2000). This study focused on HHV-6.

It was found that:Multivariate analysis showed that alternative donor (OR=3.06, p=0.038), myeloablative conditioning regimen (OR=0.22, p=0.004), and an HLA-matched (10/10 at allele level) graft (OR=0.14, p=0.011) were risk factors of HHV-6 reactivation. Interestingly, they were different from those of CMV reactivation (acute GvHD grade >=I, OR=2.74, p=0.024).HHV-6 reactivation was found earlier post-transplant than CMV reactivation (19 vs. 47 days, median values, p=0.014) and rather before clinically apparent aGvHD manifestation, in contrast to CMV and EBV infections (percentages of reactivation events prior to aGvHD were 71, 14, and 14 for HHV-6, CMV and EBV, respectively).HHV-6 was associated with an increase in CD4+ lymphocytes (p=0.028, Wilcoxon matched pairs test), in contrast to CMV infection, in which CD4+ cells were rather low.HHV-6 reactivation noted during 100 days post-transplant was associated with a higher fraction of FoxP3+ in CD4+ cells compared with those of counterparts (13% vs. 8.5%, median values, p=0.008).Encephalitis was frequent in HHV-6 (8/14 vs. 26/104, p=0.023) and EBV (9/14 vs. 22/104, p=0.002) positive patients. HHV-6 preceded EBV (14/26 vs. 28/92, p=0.037) but not CMV (9/36 vs. 33/82, p=0.145) reactivation before 100 days after HSCT.HHV-6 positivity in the 100 days post-HSCT was associated with a higher mortality rate early post-transplant (6-month survival rates of HHV-6-positive and -negative cases: 55% vs. 80%, respectively).

In conclusion: HHV-6 reactivation affects patients early post-transplant and associates with an increase in CD4+ lymphocytes in blood which frequently have FoxP3 generation potential. Risk of aGvHD is consequently lower, but there is a higher incidence of infections, including EBV, encephalitis, and sepsis, and survival is poor.

Disclosures:

No relevant conflicts of interest to declare.
Efficacy and Safety of Micafungin as An Empirical Antifungal Agent for Febrile Neutropenic Patients with Hematological Diseases.
Authors: Park Joon Seong , Kim Dong Hwan , Choi Chul Won , Jeong Seong Hyun , Choi Jin-Hyuk , Kim Kihyun , Kim Seok Jin , Jung Chul Won , Yang Deok-Hwan , Kim Hugh Chul , Jang Jun Ho
Published: October 2019

Abstract 4661

Introduction

Invasive or possible fungal infection is often fatal and its incidence is increasing in febrile neutropenic patients with hematological malignancies. Thus, empirical antifungal agent should be carefully selected for those patients.

Patients and Methods

The study was conducted as a prospective multicenter trial to document the efficacy and safety of micafungin (Mycamine®), a class of echinocandin, as an empirical antifungal agent in febrile neutropenic patients. Micafungin was administrated for sustained fever (>38.4°C) on day 3 - 5 after initiation of empirical antibiotic therapy. The overall success rate according to the composite score (no breakthrough fungal infection, survival for seven days beyond the end of therapy, did not discontinue therapy prematurely, had resolution of fever during the period of neutropenia, and successfully treated a documented base-line fungal infection) and side effects were evaluated.

Results

A total of 47 patients with AML, ALL, MDS or lymphomas were enrolled. The overall success rate was 61.7% (29/47). Three patients (6.4 %) experienced grade 3/4 elevated aspartate aminotransferase and ten patients (21 %) showed grade 3/4 hyperbilirubinemia and nine of which resolved, and four patients died of septic shock. Patients with young age (< 50 yr) and ALL rather than AML showed a better response to micafungin. Less profoundly neutropenic (≥50 /mm3) patients revealed a better response, as did the patients who recovered from fever or neutropenia.

Conclusions

Micafungin has been reported to have an excellent efficacy (61.7 %) and safety profile when used as an empirical antifungal agent to treat febrile neutropenic patients with hematological disorders.

Disclosures:

No relevant conflicts of interest to declare.
Favorable Transplantation Outcomes Associated with Early Recovery of Lymphocytes and CD8+ Cells Following Allogeneic Stem Cell Transplantation.
Authors: Moon Joon Ho , Cho Yoon Young , Kang Byung Woog , Kim Shi Nae , Lee Soo Jung , Kim Yun Jeong , Suh Jang Soo , Lee Kun Soo , Park Jae Hoo , Sohn Sang Kyun
Published: October 2019

Abstract 4665

Background

Immune reconstitution is an important component for successful transplantation. The laboratory parameters regarding immune reconstitution at 3 months after allogeneic stem cell transplantation (SCT) were analyzed in the current study.

Methods

To assess the kinetics of lymphocyte subset recovery, 128 patients who underwent allogeneic SCT and survived at least 3 months after transplantation were monitored by surface markers (CD3, CD4, CD8, CD19, CD56).

Results

Factors affecting CD4+ cell recovery at 3 months after transplantation included the use of cyclosporine instead of tacrolimus (odds ratio [OR]=0.256, p=0.007), in vivo T-cell depletion (OR=0.263, p=0.001), and use of peripheral blood stem cells instead of bone marrow (OR=0.281, p=0.016). The overall survival (OS) was better for the patients with early recovery of absolute lymphocyte counts (ANC, >1500/mm3), CD3+ (>1000/mm3), CD4+ (>300/mm3), CD8+ (>600/mm3), and CD19+ cells (>30/mm3). But CD56+ NK cell recovery (>300/mm3) failed to predict better survival. The patient group with early recovery of ANC, CD3+, and CD19+ cells showed a lower relapse rates. Treatment related mortality (TRM) was lower for the patients with early recovery of ANC, CD3+, and CD4+ cells. Chronic graft-versus-host disease (GVHD) was not related with the lymphocyte recovery. In the multivariate analysis, high risk disease status (hazard ratio [HR]=1.903, p=0.031), acute GVHD ≥grade 2 (HR=1.975, p=0.039), and HLA mismatch (HR=4.124, p=0.002) were significantly associated with lower OS, whereas, early recovery of ANC (HR=0.455, p=0.047) and CD8+ cells (HR=0.485, p=0.057) at 3 months showed a better survival.

Conclusion

Faster lymphocyte recovery and CD8+ cell recovery at 3 months after allogeneic SCT showed a better survival and lower TRM rate.

Disclosures:

No relevant conflicts of interest to declare.
Monitoring of Minimal Residual Disease in NPM1 Mutated Acute Myeloid Leukemia (AML) – a Single Center Experience in 27 Patients.
Authors: Dvorakova Dana , Racil Zdenek , Palasek Ivo , Protivankova Marketa , Jeziskova Ivana , Lengerova Martina , Mayer Jiri
Published: October 2019

Abstract 4672

Background

Mutations within NPM1 gene occurs in about 60% of adult cytogenetic normal AML (CN-AML) and represent the single most frequent molecular aberration in this subgroups of patients. These mutations usually occur at exon 12 and induce most frequently a net insertion of four base pairs.

Aims

To examine the applicability and sensitivity of DNA-based real-time quantitative polymerase chain reaction (RQ-PCR) with mutation-specific reverse primers and common minor groove binding (MGB) probe and to evaluate whether minimal residual disease levels are of prognostic relevance in CN-AML patients with NPM1 mutations.

Methods

Patients were treated within different AML trials and follow-up samples of peripheral blood or bone marrow were referred to perform an RQ-PCR. Samples were analysed at diagnosis, during, and after therapy. The NPM1 mutations were A (17 pts), B (1 pt), D (2 pts) and 7 patients with individual rare types. For all cases, levels of minimal residual disease were determined by DNA-based RQ-PCR with mutation-specific reverse primer, one common forward primer and one common MGB probe. The NPM1 mutation value was normalized on the number of albumin gene copies and expressed as the number of NPM1 mutations every 106 genomic equivalents.

This assay is highly specific as no wildtype NPM1 could be detected. Maximal reproducible sensitivity was 10 plasmide molecules per reaction.

Results

A total of 950 samples of bone marrow and/or peripheral blood from 27 patients have been analyzed. Twenty of 27 patients (74%) achieved molecular remission (MR), twenty-six of 27 patients (96%) achieved hematological remission (HR). 6 of 27 (22%) patients achieved HR without MR and one patient failed therapy. 8 of 20 patients (40%) with MR after treatment relapsed at molecular level and except one in all these patients hematological relaps occured (one patient is still in HR with bone marrow blast present, but < 5%). Considering relapsed patients, time from molecular to hematological relapse was 1 to 5 months (median: 3 months).

Considering all 14 patients with HR without MR (6 pts) or with molecular relapse (8 pts), in 11 of them hematological relaps occured (79%) and molecular positivity anticipating hematological relaps with median of 3,5 month (1-7 months). 3 of these 14 patients are still in HR.

Conclusions

Mutations within NPM1 gene are a sensitive marker for monitoring minimal residual disease in CN-AML patients. RQ-PCR using a MGB probe is an efficient approach to long-term follow-up of residual leukemia cells and frequent quantitative monitoring is useful for reliably predicting hematological relapse. Achievement of negativity appears to predict favorable clinical outcome.

This work was partially supported by research grant No. MSM0021622430

Disclosures:

No relevant conflicts of interest to declare.
Relationship Between CD56 Antigen Expression and Quantification of MDR1 Gene Expression in Patients with De Novo Acute Myeloid Leukemia(AML).
Authors: Xu Bing , Song Xiaoyan , Shi Pengcheng , Xiao Pengnan , Yu Zhengshan , Zhou Shuyun
Published: October 2019

Abstract 4677

Introdution

The expression of CD56 antigen and over-expression of the multidrug resistance gene 1 (MDR1) seems to confer the poor therapeutic outcome to AML. The aim of this study is to investigate the relationship between CD56 antigen expression and MDR1 gene expression in patients with de novo AML and explore the indicating effect of these two factors on clinical drug resistance.

Patients and methods

A real-time quantitative reverse transcriptase polymerase chain reaction method was established for detecting MDR1 expression levels and a three-color flow cytometry analysis with CD45/SSC gating was used to examine CD56 antigen expression in 79 patients with de novo AML.

Results

CD56 antigen was recorded in 19 out of 79 cases (24.1%) and particularly in those with M5 subtype and t(8;21) AML. Moreover, CD56 expression was significantly associated with unfavorable cytogenetic abnormalities (P< 0.05). Significantly higher percentage (57.1%, 4/7) of patients with t(8;21) demonstrated CD56 expression than those with favorable cytogenetic abnormalities (P< 0.05). CD56+ AML patients had higher incidence of splenohepatomegalia and level of lactate dehydrogenase than CD56- patients (P< 0.05). The median expression level of MDR1 was statistically higher in CD56+ AML patients than that in CD56- cases (P< 0.001) and 89.5% (17/19) CD56+ AML patients were found with high MDR1 expression. The CR rate in high MDR1 / CD56+ AML patients was significantly lower than that in low MDR1/ CD56- cases. (58.8% vs 89.2%, P< 0.01).

Conclusions

There is a linear correlation between MDR1 and CD56 expression in AML. This relationship may explain why CD56 expression is related to a poor prognosis in AML. Therefore both with high MDR1 expression level and CD56 antigen expression can identify AML patients with unfavorable outcome.

Disclosures:

No relevant conflicts of interest to declare.
Transforming and Tumorogenic Activity of JAK2 by Fusion to BCR: Molecular Mechanisms of Action of a Novel BCR-JAK2 Tyrosin-Kinase.
Authors: Cuesta-Domínguez Álvaro , Ortega Mara , Ormazabal Cristina , Santos-Roncero Matilde , Galán-Díez Marta , Steegmann Juan Luis , Arranz Eva , Figuera María Ángeles , Sayyah Jaqueline , Sayeski Peter P. , Vizmanos José Luis , Bueren Juan A. , Rio Paula , Fernandez-Ruiz Elena
Published: October 2019

Abstract 4683

Chromosomal translocations in human tumors frequently produce fusion genes whose chimeric protein products play an essential role in oncogenesis. Recent reports have found a BCR-JAK2 fusion gene in cases of chronic or acute myeloid leukemia, but the protein had not been characterized. We describe a BCR-JAK2 fusion gene by fluorescence in situ hybridization and RT-PCR amplification from bone marrow at diagnosis of a patient with acute lymphoblastic leukemia. After induction therapy, real time PCR showed persistent molecular response correlating with hematological remission maintained up to present. BCR-JAK2 is a 110 KDa chimeric protein containing the BCR oligomerization domain fused to the JAK2 tyrosine-kinase domain. In vitro analysis showed that BCR-JAK2 was constitutively phosphorylated and was located to the cytoplasm. BCR-JAK2 transformed the IL-3-dependent murine hematopoietic cell line Ba/F3 into IL-3 independent growth and induced STAT5b phosphorylation and translocation into the cell nuclei. The treatment with a JAK2 inhibitor abrogated BCR-JAK2 and STAT5b phosphorylation, leading to apoptosis of transformed Ba/F3 cells. To test whether BCR-JAK2 has tumorogenic ability in vivo, we performed experiments with nude mice, in which we injected subcutaneously cells transduced with the control vector and cells expressing BCR-JAK2. Notably, we only obtained tumors in the flank injected with BCR-JAK2 expressing cells, thus confirming the tumorogenic activity of the BCR-JAK2 fusion protein. We conclude that BCR-JAK2 is a new tyrosine-kinase that induces proliferation and cell survival, which can be abrogated by JAK2 inhibitors. In vitro studies demonstrate that BCR-JAK2 displays transforming activity. Moreover, the nude mice model reveals its ability to cause tumors.

Disclosures:

No relevant conflicts of interest to declare.