bluuebunny/crossref_metadata_2025_split huggingface dataset:
G-CSF Mobilized Versus Conventional Donor Lymphocytes in the Treatment of Hematological Malignancies.
Authors: Abbi Kamal K.S. , Ehmann W. Chritopher , Talamo Giampaolo , Lucas Kenneth , Carraher Michelle , Rybka Witold , Claxton David F. 
Published: October 2019

Abstract
Abstract 4301

Introduction
Donor lymphocyte infusions (DLI) can reverse relapse of hematologic malignancy following allogeneic hematopoietic stem cell transplant (HSCT) in some cases. G-CSF is used to mobilize progenitor cells for transplantation and at our center excess cells have been cryopreserved and subsequently used as DLIs. Little is known regarding the effectiveness of G-CSF mobilized cells as DLIs in the treatment of hematological malignancies. We conducted a retrospective study comparing the overall survival in patients treated with G-CSF mobilized vs. conventional DLI for hematological malignancies.

Patients and Methods
After IRB approval, we conducted a retrospective chart review of 67 patients who received DLI treatment of management of hematological malignancies.

Results
Mean age was 48.5 (range 2-78). 41 (62%) patients had AML and 26 (38%) had other hematological malignancies. Patients received 1-4 DLIs each; median was 1 DLI per patient. Median no. of CD3+ cells infused was 107/kg body weight (range 1- 265). 17 (25%) patients received a total of 32 G-CSF mobilized DLIs. 50 (75%) patients received 77 conventional DLI. 20 (29%) patients were treated with immunosuppressive agents at the time of DLI and 49 (73%) received chemotherapy in the 90 days prior to DLI. DLIs were administered for relapse or residual disease post-transplant in 62 patients. 5 patients received 7 DLIs because of poor engraftment. In total 38(56%) patients have died. Death was malignant disease related in 31/38(82%) patients, and caused by graft versus host disease (GVHD) in 5/38 (13%). GVHD deaths were 2/17 in the G-CSF and 3/50 in the conventional group (NS). Median survival after DLI was 213 days for the 67 patients and was similar for GCSF and conventional groups. 19/67 patients survived more than 1 year from the initial DLI. Death and GVHD were not predicted by diagnosis, donor type (related of unrelated), use of immunosuppressives at the time and DLI or number of infused donor CD3 cells. Time to first evidence of GVHD was similar between the two groups (p=0.35). Kaplan-Meier estimates of survival suggested that G-CSF mobilized DLI, while not statistically different from conventional DLI, showed a trend towards a survival advantage (p=0.08). This analysis is shown in the figure below.

Conclusion
In this series of poor prognosis hematological malignancies, DLI was clearly lifesaving in a number of patients. G-CSF mobilized DLI appears to be at least as effective as conventional DLI for the treatment of patients relapsing after allogeneic blood stem cell transplantation.

Disclosures:
No relevant conflicts of interest to declare.
Alternative Donor Hematopoietic Stem Cell Transplantation for Children with Severe Aplastic Anemia.
Authors: Park Meerim , Koh Kyung Nam , Bae Keun Wook , Lee Mee Jeong , Im Ho Joon , Seo Jong Jin , Moon Hyung Nam 
Published: October 2019

Abstract
Abstract 4348

Background
Hematopoietic stem cell transplantation (HSCT) from matched sibling donor is the standard first-line treatment for children with severe aplastic anemia (SAA). However, the management of SAA lacking a suitable donor remains a great challenge. For those children, HSCT using unrelated donor or mismatched related donor could be a therapeutic alternative. The purpose of this study is to evaluate the outcome in children with SAA who received HSCT from donors other than matched sibling.

Patients and Method
Between March 2003 and July 2009, 17 patients received HSCT from alternative donors (AD) at Asan Medical Center. We reviewed their medical records and analyzed their transplant-related parameters and outcome.

Results
Of a total of 17 patients, 11 were male and the median age at HSCT was 9.0 years, ranging from 3.0 to 16.7 years. Four patients had Fanconi anemia and 13 had acquired SAA including 2 who developed SAA after liver transplantation. Donors included unrelated bone marrow (U-BM) in 5, unrelated peripheral blood (U-PB) in 6, unrelated cord blood (U-CB) in 2 and related haploidentical peripheral blood (H-PB) in 4. Of 17 patients, 15 (88%) achieved sustained engraftment. Of 15 with engraftment, only 1 patient who received HSCT from U-CB died of severe GI GVHD and the other 14 patients remain on stable normal counts without transfusion support. All 2 patients (1 U-BM, 1 H-PB) who failed to engraft were dead despite DLI or 2nd HSCT. With a median follow-up of 31.9 months, the Kaplan-Meier estimated overall survival at 2 years was 76.6%.

Conclusion
In children with SAA, HSCT from AD including haploidentical family donor could be considered as a treatment option if the patients have no matched sibling donor. Given the limitation of this study such as small number of patients and short follow-up period, further trial will be necessary.

Disclosures:
No relevant conflicts of interest to declare.
Conditional Expression of Heterozygous or Homozygous Jak2V617F From Its Endogenous Promoter Induces a Polycythemia Vera-Like Disease.
Authors: Akada Hajime , Yan Dongqing , Zou Haiying , Hutchison Robert E. , Mohi M. Golam 
Published: October 2019

Abstract
Abstract 437
A somatic point mutation (V617F) in the JAK2 tyrosine kinase was found in a majority of patients with polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). However, the contribution of JAK2V617F in these three clinically distinct myeloproliferative disorders (MPDs) remained unclear. To investigate the actual role of JAK2V617F in the pathogenesis of MPDs, we generated an inducible Jak2V617F knock-in mouse, in which the expression of Jak2V617F is under control of the endogenous Jak2 promoter. Expression of heterozygous Jak2V617F evoked all major features of human PV, which included marked increase in hemoglobin and hematocrit, increased red blood cells, leukocytosis, thrombocytosis, splenomegaly, reduced serum levels of erythropoietin (Epo) and Epo-independent erythroid colonies. Homozygous Jak2V617F expression resulted in a more severe form of PV associated with markedly elevated leukocytosis, neutrophilia and thrombocytosis, and a majority of these mice succumbed due to massive cardiac thrombosis. Activation of Stat5, Akt and Erk was significantly enhanced in erythroblast cells expressing homozygous Jak2V617F compared to heterozygous Jak2V617F, suggesting that the degree of activation of downstream signaling pathways would be affected by the Jak2V617F gene dosage. This may also partly explain the severe phenotype in mice expressing homozygous Jak2V617F. We conclude that heterozygous Jak2V617F is sufficient to cause PV, whereas homozygous Jak2V617F increases the severity of PV disease and the risk of thrombosis. Our results also provide strong evidence that Jak2V617F gene dosage does not play a defining role in determining PV versus ET phenotype in this mouse model of MPD.

Disclosures:
No relevant conflicts of interest to declare.
Relationship Between Expression of TLRs and Disease Activity in CLL.
Authors: Barcellini Wilma , Zaninoni Anna , Imperiali Francesca Guia , Reda Gianluigi , Orofino Nicola , Cortelezzi Agostino , Zanella Alberto 
Published: October 2019

Abstract
Abstract 4379

Background
Toll-like receptors (TLRs) are major agents of innate immunity and initiators of adaptive immune response by acting as costimulatory signals for B cells and inducing maturation, proliferation and antibody production after pathogen recognition. They are also involved in the self-antigen recognition and could play a role in autoimmune phenomena. It is well known that chronic lymphocytic leukemia (CLL) is characterized by an increased incidence of autoimmune phenomena and immunodeficiency, which can greatly influence the disease outcome leading to a variable clinical course.

Aims
To evaluate the correlation between the gene expression of TLRs in 41 patients with CLL (mean age±SD 66±18 years, range 42-90, 15 female and 26 male), the clinical course of the disease and the expression of prognostic factors (mutational status of IgVH region, CD38 and ZAP70 expression, and cytogenetic alterations).

Methods
The gene expression of TLR4, TLR9, and TLR10 was studied. Total RNA was extracted from B cells of patients and controls (pool of 10 healthy donors), cDNA was synthesized, and real-time PCR was performed. For each reaction 50 ng of cDNA were mixed with 1.25μl of TaqMan primer/probe set and 10.25μl of TaqMan Universal Master Mix. The TLR4, TLR9, and TLR10 expression was normalized according to GAPDH as an internal control gene and it was expressed as percentage of control.

Results
TLR4 gene expression was significantly lower in CLL patients compared to controls (18±3%, mean±SE) while TLR9, and TLR10 gene expression was higher (3457±500% and 2897±440%, respectively). Moreover, considering “progressive” CLL patients (n=20), TLR4 gene expression was significantly lower compared with “indolent” ones (n=21) (10.2±2.2% vs 24.9±4.9%, p=0.01). Patients with unmutated IgVH expression and unfavorable cytogenetic alterations (mostly 11q-) showed even more reduced TLR4 gene expression, but the small numbers did not allow statistical evaluation. TLR4 gene expression was lower in patients with infective diathesis, although not significantly (11.5±2.9% vs 20.0±3.9).The 6 “progressive” CLL patients with autoimmune diseases (hemolytic anemia, Evan's syndrome and phemphigus) showed significantly higher TLR4 gene expression compared with “progressive” patients without autoimmune phenomena (17.7±4.4% vs 6.9±2.1%, p=0.02), possible expression of activated immune system.

Conclusion
The reduced TLR4 gene expression in CLL patients with “progressive” disease, unmutated IgVH status, and unfavourable cytogenetic alterations is consistent with a reduced ability to a proper immune response to Gram-negative bacterial lipopolysaccarides and consequent increased susceptibility to infections.

Disclosures:
No relevant conflicts of interest to declare.
Applicability of a New Prognostic Scoring System in Myelodysplastic Syndromes. MDS Study Group of Hematology.
Authors: Flores Gabriela , Santos Maria Isabel , Alfonso Graciela , Cabrejo Maria , Prates Maria Virginia , Iastrebner Marcelo , Fantl Dorotea Beatriz , Arbelbide Jorge , Campestri Reinaldo , Penchasky Diana , Cardenas Paula , Sackmann Federico , Garcia Juan , Nucifora Elsa , Basquiera Ana , Pavlovsky Astrid , Pavlovsky Miguel Arturo , Fernandez Isolda , Viñuales Susana 
Published: October 2019

Abstract
Abstract 4838

Background
Myelodysplastic Syndromes (MDS) are so heterogeneous, that new prognostic scoring systems are being continuously developed to help choose the best treatment strategy. The International Prognostic Scoring System (IPSS) excludes secondary MDS, prior therapy and Chronic Myelomonocytic Leukemia (CMML) with leucocytosis. Recently, a new risk model has been published by Kantarjian et al. (Cancer 2008; 113; 6; 1351) that is applicable to all MDS patients.

Aims
To validate this new risk model in our MDS population.

Methods
We analyzed 253 patients reported from 15 centers in our country from Jan 2007 through Jun 2009. We took into account age, performance status (PS), hemoglobin, platelets, leucocytes, bone marrow blasts, karyotype, and transfusion requirements. The new model divided patients into 4 prognostic groups: Low Risk (LR): 0-4; Intermediate-1 (I-1): 5-6; Intermediate-2 (I-2): 7-8; and High Risk (HR): ≥9. We assessed the prognostic impact of this New Risk Model and compared results to IPSS. Mortality Rate and AML progression risk were analyzed.

Results
164 patients were evaluable, mean age 69 (R 21-92), primary MDS: 144, and secondary MDS: 20. Risk Group assessment for the new score: LR (34%), I-1 (31%), I-2 (18%), and HR (17%). The mean Follow-up was 22 months;

Results
are shown in table and graphic.

Conclusion
Even though our sample is not considerably big, and follow-up is short, we confirmed 4 categories with different prognosis and found a close correlation with Mortality Rate. This New Scoring System allowed us to identify Low Risk IPSS patients with different outcomes, highlighted the importance of adding new prognostic factors like age and performance status, refined the cut-offs of thrombocytopenia and anemia, and recognized the adverse impact of prior transfusions needs.

Graphic

Disclosures
No relevant conflicts of interest to declare.
Removal of Autologous Activated CD4 Positive T Lymphocytes Also Result in Increased Colony-Forming Units (CFUs) in Patients with Myelodysplastic Syndromes.
Authors: Zhang Zheng , Li Xiao , Zhang Qingqiao , He Qi , Xu Feng , Wu Lingyun 
Published: October 2019

Abstract
Abstract 4862
Autologous activated CD4+ T lymphocytes (CD4+/CCR5+ double positive cells) in BMNC of 29 MDS patients were depleted or plused in in vitro cultures. That cultures with removed CD4+/CCR5+ cells exhibited significantly increased CFU generation. Conversely, the 4-fold CD4+/CCR5+ T lymphocyte-plused BMNC exhibited no colonies at all in in vitro cultures. After CD4+/CCR5+ depleted in vitro cultures, the clonal cells increased in patients with chromosome 5q– or 20q–, kept almost unchanged in patients with trisomy 8, but dramatically decreased in patients with monosomy 7. In addition, after the removal of CD4+/CCR5+ T cells, better CFUs generation were found in those patients who were with higher BM Th1 (CD4+/IFNγ+) cells or hypocellularity, or bearing DR15 allele. We concluded that selective removal of autologous activated CD4+ T-cells can increase the generation of CFUs, while the increased CFU consisting of residual normal hemopoiesis or clonal hemopoiesis kept unconcluded.

Disclosures
No relevant conflicts of interest to declare.
Stable MAGEC1/CT7 Inhibition by Short Hairpin RNA (shRNA) Does Not Affect Tumor Cell Proliferation and Invasive Potential in Sko-007 Multiple Myeloma Cell Line.
Authors: de Carvalho Fabricio , Camargo Anamaria A. , Costa Érico T. , Masotti Cibele , Andrade Valéria C.C. , Strauss Bryan E. , Caballero Otávia L. , Colleoni Gisele W.B. 
Published: October 2019

Abstract
Abstract 4888

Introduction
MAGEC1/CT7 gene encodes for a cancer testis (CT) antigen frequently expressed in multiple myeloma (MM) and may be a potential target for immunotherapy in this still incurable disease. This CT gene expression is restricted to malignant plasma cells and may be related with tumor progression, since it seems to play a role as a “gatekeeper” gene for other CT antigens and can be associated with a more aggressive phenotype. However, the exact function of this protein for tumor biology is not yet understood.

Objective
The aim of this study is to elucidate the role of MAGEC1/CT7 in the control of cellular proliferation, DNA synthesis and invasive potential of Sko-007 myeloma cell line.

Material and Methods
We used a short hairpin RNA (shRNA) specific for MAGEC1/CT7 gene that was previously inserted in the pRS (pRETROSUPER) retroviral vector. The pRS-shRNA-MAGEC1/CT7 construct was co-transfected with pCL-amphotropic packing vector in 293T cells to produce virus particles. Sko-007 myeloma cell line was transduced and selected for stable expression of shRNA-MAGEC1/CT7. Expression of MAGEC1/CT7 was analyzed by Western Blot and Real Time PCR (RQ-PCR). Functional studies included cell proliferation, 3H thymidine incorporation and invasion potential, using matrigel-coated transwell filters.

Results
Sko-007 was chosen to perform the initial functional studies since the basal MAGEC1/CT7 gene expression level was higher in this cell line when compared with the other myeloma cell lines (U266 and SK-MM-2). Sko-007wt (wild type) cell was divided into three derivatives, Sko-007ev (empty vector), Sko-007shMAGEC1/CT7Δ (antisense strand deleted – GC bases) and Sko-007shMAGEC1/CT7. MAGEC1/CT7 expression was approximately five times down regulated in Sko-007shMAGEC1/CT7 when compared with Sko-007wt, Sko-007ev, and Sko-007shMAGEC1/CT7Δ cells by RQ-PCR. Sko-007shMAGE1/CT7 had an 80% decrease in MAGEC1/CT7 protein expression when compared with Sko-007wt, Sko-007ev and Sko-007shMAGEC1/CT7Δ by Western Blot. Our preliminary functional assays did not show any change in cell proliferation and DNA synthesis when Sko-007shMAGEC1/CT7 was compared with Sko-007wt, Sko-007ev and Sko-007shMAGEC1/CT7Δ. Indeed, Sko-007 cell line has invasive potential, but no change was observed in Sko-007shMAGEC1/CT7 or other cells (Sko-007wt, Sko-007ev and Sko-007shMAGEC1/CT7Δ).

Conclusions
Expression of MAGEC1/CT7 mRNA could be inhibited by the shRNA strategy, but the concomitant reduction of its protein level did not seem to have impact in proliferation, DNA synthesis or in invasive potential of Sko-007 MM cell line (Supported CNPq and LICR São Paulo Branch).

Disclosures
No relevant conflicts of interest to declare.
Antitumor Effects and Mechanisms of Curcumin On Human Multiple Myeloma Cell Lines.
Authors: Bai Qingxian , Liu Qifa 
Published: October 2019

Abstract
Abstract 4896

BackgroundF
Multiple myeloma(MM) is a malignant plasma disease, which is characterized as high relapse rate and high resistance to chemotherapy. Curcumin is a polyphenol derived from the rhizome of Curcuma spp. It possesses diverse pharmacologic actions, such as antitumor, anti-inflammatory,anti- oxidation properties .Curcumin has the property of inhibit multiple tumor cell lines, in which included multiple myeloma cell. The real mechanism is not completely clear yet. We explored the mechanisms of curcumin on human multiple myeloma cell lines (RPMI8226 and H929), and investigated whether the combination of curcumin and adriamycin(Adr) has a synergistic effect.

MethodsF
The effect of curcumin on proliferation of RPMI8226 and H929 was observed with MTT assay. The synergetic effect of curcumin and Adr was analyzed by median-effect principle. Cell cycle distribution and apoptosis were studied with flow cytometry. Expression of surviving, bcl-2, bax mRNA was detected by RT-PCR.

ResultsF
Curcumin could inhibit the proliferation of RPMI8226 and H929 cells in a time- and dose-dependent manner. The IC50 values for RPMI8226 and H929 cell line were 12.15 μmol/L,17.24μmol/L respectively. The combination of curcumin and Adr showed synergistic effect even at low concentration of Adr. Apoptotic ratio of treated cells was significantly higher than untreated controls (36.9% vs 10.6%, p<0.05). Cells treated with curcumin showed cell cycle arrest at G2/M phase. Curcumin upregulated expression of survivin, bcl-2, while bax mRNA was significantly downregulated.

ConclusionF
Curcumin could suppress the proliferation of multiple myeloma cells and induce apoptosis. Adr combining with curcumin can show synergistic effect at low concentration of Adr. The mechanism of curcumin's antitumous effect might be related to down-regulation of surviving, bcl-2 mRNA and up-regulation of bax mRNA.

Disclosures
No relevant conflicts of interest to declare.
Localized AL Amyloidosis of the Breast: A Case Series.
Authors: Charlot Marjory , Seldin David C. , O'Hara Carl , Skinner Martha , Sanchorawala Vaishali 
Published: October 2019

Abstract
Abstract 4906
AL amyloidosis is characterized by widespread, progressive deposition of fibrillar amyloid protein derived from monoclonal immunoglobulin light chains, leading to organ failure and death. This disease is typically systemic, however, it can occur as a localized form. In localized amyloidosis, the deposits occur near the site of synthesis of the precursor protein and in some cases, plasma cells have been demonstrated histologically adjacent to the deposits. For unknown reasons, the tracheobronchial tree is the most common site for localized AL amyloidosis. Localized AL amyloidosis of the breast is a rare entity that has been described in the literature in isolated case reports. It can present as a palpable mass or as calcifications on routine screening mammography. We report here a case series of seven women (median age 63 years, range 46 to75) seen and evaluated at Boston University Medical Center from 1990-2008. We evaluated 1502 new patients with AL amyloidosis in this time period, making the incidence of localized AL amyloidosis of the breast to be 0.5% at a single referral center. All seven patients had abnormal screening mammography with calcifications, and biopsies that revealed Congo red positive amyloid deposits. Histologically, the amyloid deposits appeared as amorphous material in the stroma around the ducts and lobules in most patients; one patient had amyloid deposits in the ducts only, but not in the stroma. None of the patients had clinical or laboratory evidence of other organ involvement, all had negative Congo red staining of an abdominal fat pad aspirate, and all had a negative work up for a plasma cell dyscrasia or circulating paraprotein. The patients were treated with local excision of the regions of calcification or lumpectomy. Three out of seven patients underwent routine follow up within 6-12 months from the time of diagnosis with no evidence of disease recurrence or progression to systemic AL amyloidosis. One out of seven patients had bilateral and recurrent amyloidosis of the breasts and was found to have an associated stage I invasive ductal adenocarcinoma that was treated with lumpectomy and radiation. In summary, breast amyloidosis is rare, is not associated with a systemic plasma cell dyscrasia or amyloidosis in other organs, and can be treated surgically.

Disclosures
No relevant conflicts of interest to declare.