bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bortezomib and High-Dose Melphalan as a Novel Conditioning Regimen for Patients with POEMS Syndrome Undergoing Autologous Peripheral Blood Stem Cell Transplantation.
Authors: Tang XiaoWen , Shi Xiaolan , Hu Xiaohui , Xue Shengli , Qiu Huiying , Han Yue , Yan Lingzhi , Zhou Huifen , Liu Yuejun , Gu Bin , Sun Aining , Depei Wu 
Published: October 2019

Abstract
Abstract 4599

Background:
POEMS syndrome is a multisystem disorder associated with plasma cell dyscrasias which characterized by polyneuropathy (P), organomegaly (O), endocrinopathy (E), serum M-protein (M), and skin changes (S). Recently, we successfully treated two POEMS syndrome patients by using bortezomib-based regimen(VDD) and followed by autologous peripheral blood stem cell transplantation (APBSCT) with bortezomib combined with high-dose melphalan(Mel) conditioning regimen.

Methods:
According to the latest Mayo Clinic criteria, two patients’ diagnosis of POEMS syndrome could be comfirmed. Both of them had no response to the classical treatments, but achieved near complete remission(CR) after 4 cycles and 1 cycle of VDD respectively(Bortezomib 1.3–1.6 mg/m2/w×4 weeks; 40 mg of Liposomal doxorubicin on the fourth day of the first week; and 10 mg of dexamethasone during the initial 4 days of first cycle. Each course was at 21 days interval). Then, autologous peripheral blood stem cell collection was performed sucessfully (2.16×106/kg and 3×106/kg respectively) after mobilization by cyclophosphamide (3g/m2/d for 1 day) with subcutaneous G-CSF. APBSCT were performed approximately 1 month after stem cell collection. Two patients were conditioned with Bortezomib 1.6mg/m2(d-13,d-6,d+1,d+7)+Mel 200mg/m2(d-3)and Bortezomib 1.3 mg/m2(d-8,d-1,d+6,d+13)+ Mel 200mg/m2(d-2) respectively.

Results:
No toxic death or serious adverse effects occurred during APBSCT. Neutrophil and platelet recoveried at +10d, +27d and +7d, +20d respectively. Only case 1 patient developed mild hypoxemia during neutrophil engraftment period, but was successfully treated with corticosteroid and antibiotic therapy. The case 2 presented mucositis, diarrhea and nausea during neutropenic peroid. A follow-up period were 12 and 6 months respectively. Both of them achieved continuous CR. There was a dramatic improvement in clinical symptoms and serum VEGF levels in all two patients post VDD treatment including organomegaly (splenomegaly), M-protein, pericardiac/pleural effusion,ascites and polyneuropathy.

Conclusion:
This is the first report of POEMS patients treated by APBSCT with bortezomib and high dose Melphalan conditioning regimen. Our results suggest that Bortezomib is a new effective and relative safe therapeutic option for POEMS syndrome not only in the conventional treatment but also in APBSCT procedure.

Disclosures:
No relevant conflicts of interest to declare.
Epigenetic Downregulation of Secreted Frizzled-Related Proteins In Philadelphia Positive and Philadelphia Negative Myeloproliferative Neoplasms
Authors: Bennemann Karla , Schubert Claudia , Wilop Stefan , Brummendorf Tim H. , Galm Oliver , Jost Edgar 
Published: October 2019

Abstract
Abstract 4647

Introduction:
Hypermethylation of CpG islands in promoter regions is associated with transcriptional inactivation and represents an important mechanism of tumor suppressor gene silencing and plays a role in the pathogenesis of hematopoietic malignancies. Myeloproliferative neoplasms (MPN) constitute a heterogenous group of hematopoietic malignancies and comprise chronic myeloid leukemia (CML), polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). BCR/ABL-negative MPN (Ph-MPN) are often characterized by an autoactivating mutation in the JAK2 tyrosine kinase (JAK2V617F) as well as additional mutations in the JAK-STAT pathway. Secreted Frizzled-related proteins (SFRPs) are antagonists of the Wnt signaling pathway. Consistent with the central role of the Wnt pathway for stem cell maintenance and differentiation of hematopoietic progenitors. Upregulation of Wnt-pathway has been described in haematopoietic malignancies and epigenetic downregulation of SFRPs has been shown in acute leukemia.

Methods:
We determined the promoter methylation status of SFRP1, -2, -4 and -5 in human cell lines and in 60 samples from MPN patients by methylation-specific PCR (MSP). The JAK2V617F mutation was assessed by allele-specific PCR.

Results:
Aberrant methylation among primary MPN samples was 5 % (3/60) for SFRP-1, 24 % (14/60) for SFRP-2, 2% (1/60) for SFRP-4 and 0% (0/60) for SFRP-5. The SFRP-2 promoter was methylated in 0% of CML (0/12), 27% of PV (3/11), 30% of ET (3/10) and 26% of PMF patient samples (7/27). There was no correlation between SFRP-2 hypermethylation and MPN subtype. We detected a trend for a correlation between methylation of the SFRP-2 promoter and presence of JAK2V617F mutation (p = 0,1).

Discussion:
Our data indicate that epigenetic dysregulation of the Wnt signaling pathway is a common event in Ph-MPN with aberrant methylation of at least one SFRP being detected in 30% of the primary patient samples with the most predominant member of the family, SFRP2 being methylated in about one third of the cases. Aberrant methylation of SFRPs may contribute to the pathogenesis of MPN by influencing stem cell maintenance and proliferation of hematopoietic progenitors.

Disclosures:
No relevant conflicts of interest to declare.
Anaphylactic Reaction After Replacement Therapy In Haemophilia B Patient with Inhibitor to Factor IX
Authors: Scaraggi Rosanna , Giordano Paola , Ettorre Cosimo Pietro , Malcangi Giuseppe , Marino Renato , Lassandro Giuseppe , De Mattia Domenico , Scaraggi Francesco Antonio 
Published: October 2019

Abstract
Abstract 4662
Replacement therapy is a very hard challenge in haemophilia B with inhibitor. We describe the case of a child with severe haemophilia B and with a family history positive for development of inhibitor to factor IX (FIX) and for occurrence of allergic reaction after replacement therapy. Genetic analysis demonstrated an almost complete deletion of FIX gene. The child received replacement therapy first in his life when he was 5 years old because of the occurrence of a large haematoma of the left thigh. He was treated with recombinant FIX concentrate at the dosage of 30 IU/kg daily for three days and no inhibitor to FIX was evidenced after this therapy. Two months later the patient was treated with a single dose of recombinant FIX concentrate at the same dosage for the occurrence of a post-traumatic joint bleeding of the right knee. An inhibitor towards FIX (1.7 B.U.) was detected two weeks after this treatment and confirmed in a subsequent analysis performed after ten days (1.4 B.U.). One month later the patient was hospitalized for a post traumatic hemarthrosis of the right shoulder. In this occasion it was planned to treat the patient with recombinant FIX under careful monitoring in intensive care unit considering of the inhibitor. After the slow endovenous infusion of 200 IU of recombinant FIX concentrate we stopped immediately the administration because the child presented cough, mild respiratory failure, tachycardia, tongue and lips oedema, lips cyanosis, diffuse vasodilatation, psychomotor agitation. He received also hydrocortisone, antihistaminic by intramuscular injection and oxygen by facial mask. The presence of inhibitor towards FIX and anaphylactic reaction occurrence preclude forever any replacement therapy with FIX both recombinant and plasmatic (PCC and/or aPCC). After this episode the patient needed another hospitalization for a tonsil bleeding with severe anaemization. We treat him with recombinant activated FVII (rFVIIa) first at the dosage of 270 mcg/kg in bolus, and after at the dosage of 90 mcg/kg every three hours for one day with complete bleeding remission. We encourage the careful monitoring of inhibitor towards FIX in haemophilic B patients especially with large FIX gene deletion. It could prevent severe anaphylactic reaction during replacement therapy. Considering the previous anaphylactic reaction, in this child rFVIIa represents the only therapeutic option for bleeding management.

Disclosures:
No relevant conflicts of interest to declare.
Essential Role for Mir-181a1/b1 In T-Cell Acute Lymphoblastic Leukemia
Authors: Fragoso Ana Rita , Mao Tin , Wang Song , Schaffert Steven , Min Hyeyoung , Pear Warren S. , Chen Chang-Zheng 
Published: October 2019

Abstract
Abstract 470
MiRNA-mediated gene regulation represents a fundamental layer of post-transcriptional control of gene expression with diverse functional roles in normal development and tumorigenesis. Whereas some studies have shown that over-expression of miRNA genes may contribute to cancer development and progression, it is yet to be rigorously tested by the loss-of-function genetic approaches whether miRNA genes are required for cancer development and maintenance in mice. Here we show that mir-181a1/b1 coordinates Notch and pre-TCR signals during normal thymocyte differentiation and plays an essential role in development and onset of T-cell acute lymphoblastic leukemia (T-ALL) induced by some Notch mutations. Using gain-of-function and loss-of-function approaches, we demonstrated that mir-181a1/b1 controls Notch and pre-TCR receptor signals during the early stages of T cell development in the thymus by repressing multiple negative regulators of both pathways, including Nrarp, PTPN-22, SHP2, DUSP5, and DUSP6. These results illustrate that a single miRNA can coordinate multiple signaling pathways by modulating the timing and strength of signaling at different stages. Intriguingly, synergistic signaling between Notch and pre-TCR pathways is necessary for the development of T-ALL, and miR-181 family miRNAs are aberrantly expressed in T-ALL patients. These observations raise the possibility that mir-181a1/b1 might contribute to the onset or maintenance of T-ALL by targeting similar pathways in tumor cells as it does in normal thymic progenitor cells. In support of this notion, we found that loss of mir-181a1/b1 significantly delayed the onset and development of T-ALL induced by intracellular domain of Notch1 (ICN1) and caused a 32% increase in the median survival time from 41 days to 54 days in T-ALL mice. Importantly, we noted that loss of mir-181a1/b1 more efficiently repressed the leukemogeneic potential of cells with lower levels of ICN1 expression, suggesting that mir-181a1/b1 may be more effective in inhibiting T-ALL development induced by a Notch mutant with weaker signal strength. Indeed, we demonstrated that loss of mir-181a1b1 essentially blocked T-ALL development induced by the weaker Notch mutant and dramatically decreased mortality from 60% to 10% in these T-ALL mice. Since human Notch mutations identified in T-ALL patients generally have weaker signaling strength and lower oncogenic potential than that of ICN1, our findings indicate that mir-181a1/b1 may play an essential role in development of normal thymic progenitors and Notch-induced T-ALL and may be targeted to treat T-ALL patients harboring Notch mutations.

Disclosures:
No relevant conflicts of interest to declare.
Incipient Release of Leukocytes After Allogeneic Hematopoietic Cell Transplantation Is Indicated by Increasing Serum Levels of Uric Acid Before Detection of First Leukocytes
Authors: Haen Sebastian P. , Eyb Vicky , Kanz Lothar , Bethge Wolfgang A. 
Published: October 2019

Abstract
Abstract 4701
Uric acid serves as an endogenous danger signal to the immune system upon the release from dying cells activating both innate and adaptive immunity. Multiple cytokines and danger signals are released into the blood modulating cellular and humoral immune responses during haematopoietic cell transplantation (HCT). Here, we investigated serum levels of uric acid during haematopoietic regeneration after allogeneic HCT to evaluate whether uric acid can be used as marker for hematopoietic engraftment indicating immunologic activity below the detection limit of differential blood counts (50/μ l). Differential blood counts and serum levels of uric acid, creatinine and urea were retrospectively analyzed in 47 consecutive patients (22 men and 25 women, median age 48 years, range 19 to 73 years) during 50 allogeneic HCT (three patients received a second transplantation due to recurrent disease). Serum levels of uric acid, creatinine and peripheral blood counts were determined daily. All patients received allopurinol for 5 days from the beginning of the conditioning which was discontinued afterwards. The mean nadir in uric acid levels was observed 13.4 days after the last dose of allopurinol (range 2 to 32 days) and 9.1 days after transplantation (range 2 to 21 days) irrespective of the conditioning regimen used. In all patients a significant decrease of uric acid serum levels after the transplantation was observed (Student's t-test, p < 0.001). A re-increase of uric acid was observed 1 to 10 days (mean 4.1 days, SD 2.4) before detection of white blood cells (leukocytes > 50/μ L) in 35 out of 50 transplantations (70%, group 1). In three cases (6%), the increase of uric acid levels was observed concomitantly with increasing leukocytes (group 2). In the remaining 12 cases, this elevation was only observed later (1 to 6 days after re-increase of leukocytes, mean 2.8 days, SD 1.5, group 3). The difference between group 1 and 3 was statistically significant (Student's t-test, p < 0.001). No correlation with patient age, conditioning regimen, HLA compatibility or additional medications could be observed. Therefore, a re-increase of uric acid can be indicative for incipient haematopoietic engraftment after allogeneic HCT and may serve as indicator for consumption of the ealiest differentiated cells in peripheral blood even before leukocytes can be detected.

Disclosures:
No relevant conflicts of interest to declare.
Studies on Preparation of Artesunate-Loaded mPEG-PLGA-Nanoparticles and Its Inhibition on K562 cells
Authors: Xiaoli Liu , Wang Shuang , Du Qingfeng , Xu Na , Yang Jun , Wang Yu , Zhang Jinfang , Xu Lulu , Li Xiang , Li Guofeng 
Published: October 2019

Abstract
Abstract 4712
The aim of the present work was to investigate the characterization of Artesunate-loaded methoxy poly (ethylene glyeol) -poly (lactic-co- glycolic acid) copolymer (mPEG-PLGA) nanoparticles and the anti-tumoral activity of the Art-Nps on human leukemia cells K562. The Artesunate (Art) poly (ethylene glyeol-lactic-co-glycolic acid) nanoparticles (Art-Nps) were prepared by modified-Spontaneous emulsion solvent diffusion method. The shape of the nanoparticles was observed by SEM. The mean diameter and the size distribution of nanoparticles were determined by laser light scattering. The drug loading efficiency, encapsulation rate and releasing behavior of Art -NPs in vitro were examined by HPLC. The effects of Art-Nps on the proliferation of K562 cells were studied by MTT assay and Hoechst staining. Artesunate loaded mPEG-PLGA nanoparticles were spheric with the mean size of (156.7±1.01)nm, zeta potential was -(26.23±1.86) mV, and the average drug loading and encapsulation efficiency were (14.5±0.2)% and (86.5±0.5)%, respectively. In vitro release behavior could be described by the Higuchi equation: Q=4.11t1/2+27.05, r=0.983. MTT assay showed different concentrations. different times of Art-Nps could inhibit the proliferation of K562 cells (P<0.01), and both have a synergistic effect (P=0.002), it showed that concentration was dependented with time, and the inhibition rate after 72h was exceeded to the the control group (P<0.05), it was showed the Art-Nps had sustained-release effect. Art-NPs of the cells (treated by 12.5μ g/m. 25μ g/ml. 50μ g/ml) resulted in significantly higher apoptosis than blank groups. The Art-NP obtained were characted with a small size and high drug loading and entrapment efficiency, in vitro release showed a good sustained-release nature, and it can inhibit the proliferation of human leukemia K562 cells in vitro, extending the time on leukemia cells. This study was provide an experimental basis for develop a new intravenous artesunate formulations.

Disclosures:
No relevant conflicts of interest to declare.
A Single Dose Filgastrim In the Treatment of Chemotherapy Induced Neutropenia
Authors: Jagarlamudi Kuppuswamy , Ahluwalia Meena , Patel Jay , Patel Arunbhai G. 
Published: October 2019

Abstract
Abstract 4774

Background:
Chemotherapy induced neutropenia (CIN) was a challenge to physicians until the discovery of growth factors. Neutropenia predisposes the patient to infection with increased morbidity, mortality, and cost of hospitalization. Not only does CIN leads to a delay in chemotherapy but also a dose reduction, which results in decreased efficacy and impact on survival. Filgrastim is a granulocyte colony stimulating factor that leads to the proliferation, differentiation and function of neutrophils. With an increase in the neutrophil count and function, Filgrastim reduces the incidence of neutropenia and the complications associated with febrile neutropenia (FN), making it possible to deliver chemotherapy on time, without dose adjustment. ASCO guidelines recommend that Filgastrim be given at 5 mcg/kg/day starting from days 1–3 after chemotherapy for patients with the risk of FN greater than 20% until the absolute neutrophil count is near-normal. It was recently reported that two doses of Filgrastim were comparable to higher number doses to prevent the nadir. We hypothesized that only a single dose is sufficient to prevent the nadir and delay in treatment.

Patients and Methods:
A retrospective analysis of chart data from all patients treated at The Brooklyn Hospital Center ambulatory clinic was reviewed after the initiation of chemotherapy. Data collected included the white cell count, neutrophil count, dates of chemotherapy and Filgrastim administration. Patients who received one dose of Filgastrim within 2–3 days, but not less than 24 hours prior to receiving subsequent chemotherapy comprised the study population. Data was analyzed for each individual cycle of chemotherapy.

Results:
The study population consisted of 11 patients with a total of 33 cycles of chemotherapy. Of the 11 patients, 9 had breast cancer, 1 had embryonal cancer, and 1 had colon cancer. Chemotherapy varied from Etoposide and Cisplatin, FOLFOX with Bevacizumab, and Adriamycin with Cyclophosphamid (AC) followed by Paclitaxel where the AC was given as a dose dense regimen. All the cycles had adequate response in neutrophil count to a single dose of Filgrastim when it was delivered within 2–3 days, but not before 24 hours of receiving chemotherapy. The mean absolute neutrophil count prior to Filgrastim was 0.8 compared to 6.0 (p=0.0001) after a single dose. There was no delay in treatment noticed as a result of the single dose of Filgrastim in all the 33 cycles.

Conclusion:
Filgrastim as a single dose is sufficient to treat CIN and prevent delay in subsequent treatment.

Disclosures:
No relevant conflicts of interest to declare.
Lineage-Specific Sox7 Expression In Hematopoietic Progenitor Cells Derived From Human Umbilical Cord Blood
Authors: Wan HaiXia , Chow Howard C.H. , Fung Tsz-Kan , Fan Kin-Pong , Wong Anders S.Y. , Liang Raymond , Leung Anskar Y.H. 
Published: October 2019

Abstract
Abstract 4781

Introduction:
The SOX (Sry-related HMG box) genes belong to a family of transcription factors containing a High-Mobility-Group box domain. In an initial screen of SOX genes in human leukemias, SOX7 is uniquely down-regulated in acute myeloid leukemia, myelodysplastic syndrome and chronic myelogenous leukemia but up-regulated in most cases of acute lymphoblastic leukemia. The observation led to the proposition that SOX7 may play a role in lineage differentiation in hematopoiesis. In this study, we examined SOX7 expression in human umbilical cord blood (UCB) with a view to understand its role in hematopoietic lineage specification.

Methods:
Mononuclear cells (MNC) were isolated from UCB and fractionated into CD34+, CD34-, CD34+CD38+ and CD34+CD38- populations by immunomagnetic selection and fluorescence activated cell sorting (FACS). 0.1–0.4 × 106 CD34+ UCB cells were transplanted intravenously into sub-lethally irradiated NOD/SCID mice with or without prior anti-CD122 antibody injection. Donor engraftment was assessed after 7–8 weeks and engrafting human cells were sorted for CD34+, CD33+, CD19+ expression. CD34+ UCB cells were also plated in colony-forming unit (CFU) assay. SOX7 expression was evaluated by quantitative real-time reverse-transcriptase polymerase chain reaction (Q-RT-PCR).

Results:
The relative SOX7 expression in CD34+ and CD34+CD38- UCB cells was 9.15±2.18 and 4.53±1.09 fold higher than in CD34- (P=0.0001) and CD34+CD38+ (P=0.014) respectively. SOX7 expression in CD34+ cells (arbitrarily set as 1.0) was down-regulated upon differentiation in CFU assay (CFU-GM/G: 0.09±0.06 fold, P=0.007; BFU-E: 0.05±0.03 fold, P=0.003). CD34+ cells engrafted into NOD/SCID mice as enumerated by human CD45+ cells in mouse BM (with anti-CD122: 51±6.3%; without anti-CD122: 28±9%; p=0.017) and gave rise to predominantly B-lymphoid (CD19+) (53.1±8.18%) and to a less extent myeloid (CD33+) (2.9±0.54%) differentiated cells. Prior injection with anti-CD122 antibody had no effect on lineage differentiation and the results were pooled. When SOX7 expression in engrafting myeloid and B-lymphoid differentiated cells (arbitrarily set as 1.0) were compared, it was significantly down-regulated in the myeloid series (0.28±0.08 fold, p = 0.036).

Conclusion
SOX7 is selectively expressed in human HSC from UCB. It is preferentially down-regulated during myeloid differentiation both in-vivo and in-vitro. The function of SOX7 during lineage specification and its link to myeloid malignancies is currently investigated in our laboratory.

Acknowledgments
The project was supported by a grant from the strategic Research Theme of cancer stem cells in the HKU.

Disclosures:
No relevant conflicts of interest to declare.
Differential Regulation of Expression of the Anti-Leukemogenic Orphan Nuclear Receptor Nur77 (nr4a1) In HSCs, CLPs, and CMPs
Authors: Shin Yusup , Manjunath Shwetha , Gross Matthew , Land Ruben , Garrett Russell , Emerson Stephen G. , Punt Jenni A. 
Published: October 2019

Abstract
Abstract 4782
The molecular basis for the balance between self-renewal and differentiation that regulates hematopoietic stem cell (HSC) development is poorly understood. Recent studies suggest that levels of expression of the orphan nuclear hormone receptor Nur77 (nr4a1), previously implicated in thymic selection, may play a pivotal role in HSC fate decisions. Previously published quantitative PCR data suggest that murine HSCs express relatively high levels of Nur77 mRNA. Genetic deletion of Nur77 and its close family member Nor-1 (nr4a3) in mice resulted in the development of acute myeloid leukemia (Mullican et al. Nat Med, 13:730;2007), consistent with Nur77's role as an anti-leukemogenic transcription factor, but also suggestive of a role in HSC homeostasis. To more closely investigate the role of Nur77 in normal hematopoiesis, we constructed a transgenic reporter mouse in which GFP is driven by the Nur77 promoter (eGFPNur77). Unexpectedly, we found that fewer than 3% of freshly isolated bone marrow HSCs (Lin-c-Kit+Sca-1+, IL-7R-) and common lymphoid progenitors (CLP = Lin-c-Kit+Sca-1+, IL-7R+), and no common myeloid progenitors (CMP) expressed Nur77 in our novel reporter system. Interestingly, culture of Lin- bone marrow cells with serum-containing medium resulted in a marked increase in Nur77 (GFP) expression among the majority of HSC's and CLP's, but not CMP's. Further studies indicate that a heat-insensitive component of serum, also present in plasma, is responsible for this induction. Taken together with published data, our studies suggest that induction and modulation of Nur77 may play a previously unappreciated role in self-renewal and lymphoid versus myeloid lineage commitment decisions of HSCs. We are currently working to understand the developmental difference in HSC/CLP/CMP responsiveness and to identify the serum component(s) responsible for upregulating Nur77.

Disclosures:
No relevant conflicts of interest to declare.