bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lung-Derived Microvesicles Induce Stable Long-Term Epigenetic Changes In Marrow Cells
Authors: Aliotta Jason M , Pereira Mandy , Amaral Ashley , Dooner Mark S , DelTatto Michael , Papa Elaine , Goldberg Laura R , Quesenberry Peter J 
Published: October 2019

Abstract
Abstract 4799
We have previously shown that microvesicles derived from irradiated murine lung cells enter murine bone marrow cells and induce expression of the pulmonary epithelial cell-specific mRNAs Surfactants A-D (Sp-A-D), Aquaporin-5 (Aq-5) and Clara Cell Specific Protein (CCSP). The early mRNA increases are due to direct delivery of mRNA and due to the transfer of a transcription factor to marrow cells (Aliotta JM, et al, Exp Hematol, 2010). The modulated marrow cells produce pulmonary epithelial cell-specific protein and have an enhanced ability to convert into pulmonary epithelial cells after transplantation into lethally-irradiated mice. In the present studies, we evaluated the stability of pulmonary epithelial cell-specific mRNA expression in cytokine-stimulated liquid culture and after in vivo transplantation into lethally-irradiated mice. Whole bone marrow cells isolated from C57BL/6 mice were cultured for seven days opposite C57BL/6 lung fragments, but separated from them by a 0.4 micron cell-impermeable membrane. Marrow cells were then either infused into lethally-irradiated C57BL/6 mice or established in culture with DMEM supplemented with IL-3, IL-6, IL-11 and steel factor. Engrafted mice were sacrificed six weeks after transplantation and real-time RT-PCR was carried out on marrow, liver, spleen and thymus tissue. Expression of Sp-A, B and D, CCSP and Aq-5 were variably seen in all tissues and increases over control ranged from 12-fold to 58-fold. Liquid culture was analyzed every two weeks out to 12 weeks and high levels of expression of different pulmonary epithelial cell-specific mRNAs and proteins (by western blot and Immunohistochemical staining) were observed. In order to determine the mechanism for the increased expression in culture, we established co-cultures of Fischer-344 rat lung opposite C57BL/6 mouse bone marrow cells and, using species-specific primers, determined whether the increased pulmonary epithelial cell-specific mRNA expression at each time point of secondary cytokine-supported culture was rat or mouse-specific. In every instance, the pulmonary epithelial cell-specific mRNA which was detected was mouse-specific, not rat-specific, and thus the long-term expression of lung-specific mRNA in mouse marrow was due to induced transcription by the mouse marrow cells. These data indicate that lung-derived microvesicles induce long-term stable epigenetic changes in marrow cells and suggests that the genetic changes induced by tissue microvesicles in neighboring cells may be biologically important.

Disclosures:
No relevant conflicts of interest to declare.
Long Term Results of Stanford V Regimen and Highly Active Antiretroviral Therapy (HAART) In 59 Patients (pts) with HD and HIV Infection (HD-HIV)
Authors: Spina Michele , Gabarre Jean , Mancuso Salvatrice , Re Alessandro , Schiantarelli Clara , Talamini Renato , Vaccher Emanuela , Tirelli Umberto 
Published: October 2019

Abstract
Abstract 4827

Background:
The introduction of HAART has significantly improved the outcome of pts with HD-HIV. However there are no data on the long term follow-up of HD-HIV pts treated with conventional chemotherapy (CT) regimens. In 2002, we reported the results of a prospective phase II study with the intensive 12-week CT with adjuvant radiotherapy (Stanford V) and concomitant HAART in 59 pts (Spina et al. Blood 2002;100:1984-1988).

Methods:
To analyze the long term outcome of patients included in the Stanford V and HAART protocol.

Results:
The median follow-up is 67months (range 3–156 months The 5-yr overall survival (OS), freedom from progression (FFP), disease free survival and event free survival are 54%, 52%, 60% and 37%, respectively. The 5-year OS is significantly different in pts with an international prognostic score (IPS) >2 in comparison to that of pts with an IPS <3 (84% vs 36%, p= 0.0005). Similarly, the percentages of FFP at 5 years in these groups are 72% and 45% (p= 0.03).

Conclusions:
Our data confirm the long term efficacy of Stanford V regimen in combination with HAART in HD-HIV. However, Stanford V is significantly less effective in pts with IPS>2 and therefore new strategies be tested in this setting.
Supported by AIRC and ISS grants.

Disclosures:
No relevant conflicts of interest to declare.
Deletion of PAX5 Is Frequently Accompanied by CDKN2A Deletion but Not by ETV6-RUNX1 Rearrangement In B-Lineage Acute Lymphoblastic Leukemia
Authors: Kim Miyoung , Jeon Young Jin , She Cha Ja , Hwang Sang Mee , Kang Hyoung Jin , Shin Hee Young , Ahn Hyo Seop , Yoon Sung-Soo , Kim Byoung Kook , Park Myoung Hee , Lee Dong Soon 
Published: October 2019

Abstract
Abstract 4841

Introduction
The PAX5 gene (Paired box gene 5) works as a guardian in B-lymphopoiesis. It activates essential components of B lymphopoiesis and represses transcription of genes for differentiation to other lineage. Thus, abrogation of normal function of PAX5 is believed to contribute the leukemogenesis in B-ALL. In this study, we evaluated the utility of the detection of PAX5 gene alteration in practice in relation to PAX5 immunoexpression, other common cytogenetic changes and survival in childhood and in adult B-ALL.

Materials and Methods
Interphase FISH using PAX5 split signal probe (DakoCytomation, Glostrup, Denmark), and probes for BCR-ABL1, ETV6-RUNX1 and MLL rearrangements, and CDKN2A deletion was performed in 112 CD19+ B-ALL patients (79 children, 33 adults). G-banding and immunohistochemical staining with anti-PAX5 (BC/24, BIOCARE MEDICAL, CA, USA) antibody was also performed.

Result
The incidence of translocation, deletion and amplification was 2.5%, 10.0% and 12.7% in children, and 0.0%, 18.2% and 6.1% in adults, respectively. Alteration of PAX5 gene was more common than BCR-ABL1 and MLL rearrangement in children (8.9% and 5.1%, respectively), and MLL rearrangement in adults (3.1%). None of the children with ETV6-RUNX1 rearrangement (22.8%) showed PAX5 gene alteration. Most cases of the deletion of PAX5 were accompanied by CDKN2A deletion. Positive immunoexpression in PAX5 IHC was observed in 74.4% in children and in 76.9% in adults Either the children with positive or negative PAX5 immunoexpression had PAX5 gene abnormalities. The presence of PAX5 gene alteration or negative immunoexpression of PAX5 did not have prognostic impact.

Discussion
Based on high incidence of structural alteration of PAX5 in dominant leukemic clone, we suggest PAX5 FISH as a useful tool for the diagnosis and the disease monitoring in B-ALL. Frequent CDKN2A deletion, but not ETV6-RUNX1 rearrangement with structural alteration of PAX5 implies the role of PAX5 in determining prognosis of B-ALL. No definite relationship between the structural alteration and immunoexpression of PAX5 might be resulted from allele specific regulation and haploinsufficiency.

Disclosures:
No relevant conflicts of interest to declare.
Analysis of Genetic Alterations In 843 Korean Acute Leukemia Patients by Multiplex RT-PCR Assay (HemaVision) and Karyotyping
Authors: Lim Jihyang , Kim Myungshin , Kim Yonggoo , Han Kyungja , Kim Hee-Je , Lee Seok , Min Woo-Sung , Chung Nak-Gyun , Cho Bin , Kim Hack-Ki 
Published: October 2019

Abstract
Abstract 4845

Background:
The importance of molecular tests has been increased for diagnosis, classification and MRD monitoring in acute leukemia. Recently, multiplex RT-PCR assays (HemaVision) for simultaneous detection of 28 leukemia-associated translocations was introduced and has been used for molecular screening of leukemic patients. We analyzed the results of multiplex RT-PCR assay (HemaVision) and karyotyping in Korean acute leukemia patients.

Methods:
From April 2007 to March 2010, multiplex RT-PCR assay (HemaVision, DNA Technology A/S, Denmark) was performed in 843 Korean acute leukemia cases (AML 566 vs. ALL 277 and adults 596 vs. pediatrics 237) for screening leukemia-associated translocations. All results were compared with chromosome analysis results by conventional karyotyping.

Results:
Three hundreds fifty-one cases (41.6%) were positive in multiplex RT-PCR. In AML cases, 221 cases (39.0%) revealed positive results and RUNX1-RUNX1T1 85 cases (38.5%), PML-RARA 74 cases (33.5%), CBFB-MYH11 25 cases (11.3%) and MLL-related 19 cases (8.6%) were frequently detected. In contrast, 130 cases (46.9%) revealed positive results in 277 ALL cases and BCR-ABL1 68 cases (52.3%), ETV6-RUNX1 28 cases (21.5%), TCF3-PBX1 15 cases (11.5%), MLL-related 10 cases (7.7%) were frequently detected. In comparison to karyotyping, 337 cases (96.0%) revealed abnormal karyotype and 14 cases (4.0%) revealed normal karyotype in 351 cases that revealed positive results by multiplex RT-PCR assay. In 492 cases that revealed negative results by multiplex RT-PCR assay, 278 cases (56.5%) revealed abnormal karyotype and 214 cases (43.5%) revealed normal karyotype. Overall 629 cases (74.6%) were detected genetic alterations by multiplex RT-PCR assay and/or karyotyping.

Conclusion:
Most acute leukemias may have a genetic alteration in their leukemogenesis. Both multiplex RT-PCR assay and karyotyping are essential for screening leukemia-associated genetic alterations.

Disclosures:
No relevant conflicts of interest to declare.
Chromatin Texture and Molecular Features Are Independent Prognostic Factors In AML
Authors: Mello Mariana Rezende Bandeira , Albuquerque Dulcinéia Martins , Albanez Krizzia Borges , Adam Randall L. , Pereira-Cunha Fernanda Gonçalves , Pagnano Katia B. , Costa Fernando Ferreira , Metze Konradin , Lorand-Metze Irene 
Published: October 2019

Abstract
Abstract 4850

Introduction:
The WHO 2008 classification of acute myeloid leukemia (AML) is based on morphology, cytogenetics and molecular features. Among them, mutations and internal tandem duplications of FLT3 in AML with a normal karyotype have been associated to a poor prognosis. Mutated NPM1 in the absence of FLT3-ITD is associated to a favorable. On the other hand, variables of nuclear chromatin texture have been described as independent risk factors in several malignancies (ALL, melanoma and multiple myeloma).

Aim:
To compare the influence on overall survival of the chromatin fractal dimension and molecular features in adult patients with AML.

Patients and Methods:
We analyzed 106 consecutive cases diagnosed at our Institution between 2007 and 2009. Diagnosis was made by bone marrow (BM) cytology and karyotype, and cases were classified by WHO criteria. Genomic DNA was extracted by phenol-chloroform. Genotyping was made with the MegaBACE 1000 equipment and analyzed in the Fragment Profiler v1.2. For detection of the FLT3-TKD mutation, genomic DNA was amplified by PCR followed by restriction analysis. Blasts from the diagnostic BM cytology were digitalized, segmented and nuclear morphometric variables were examined. Their influence on overall survival was analyzed in the Cox model.

Results:
Median age: 52 years; peripheral blood (PB) leukocytes: 24.0×109/l (0.7-281.3). In the univariate analysis were significant: PB leukocyte count (p=0.005), low-risk karyotype (p=0.002), FLT3 ITD+ (p=0.002), FLT3+NPM1- (p=0.029) and “goodness of fit” (R245) of the chromatin fractal dimension (Minkowski) (p=0.03). Age, fractal dimension and methylation status of p15, p16, p57, p73, ER and MDR1 were not significant. In the multivariate analyses including age, PB leukocytes, R245 and mutations, high leukocyte counts (p=0.03) and low R245 (p=0.01) were independent unfavorable and FLT3-NPM1- (p=0.04) and FLT3-NPM1+ (p=0.02) were favorable prognostic variables.

Conclusions:
The blast chromatin texture measured by R245 was an independent prognostic factor together with known risk variables in AML.
Supported by: FAPESP and CNPq

Disclosures:
No relevant conflicts of interest to declare.
Genetic Alterations of Wilms' Tumor 1 (WT1) Gene In Korean Patients with Normal Karyotype Acute Myeloid Leukemia
Authors: Choi Yunsuk , Lee Je-Hwan , Lee Jung-Hee , Kim Dae-Young , Kim Sung-Doo , Lim Sung-Nam , Hur Eun-Hye , Seoul Miee , Lee Young-Shin , Kang Young-Ah , Jeon Mijin , Lee Kyoo-Hyung 
Published: October 2019

Abstract
Abstract 4852

Introduction:
Acquired mutations of the WT1 gene have been reported in 5–10% of normal karyotype AML (NK-AML) patients. Poor prognostic impact of the mutations has been reported, but some studied found no such correlation. Recent report from Germany showed the association of single nucleotide polymorphism (SNP) rs16754 status with clinical outcomes in NK-AML (J Clin Oncol 2009; 28:578). We assessed clinical implication of WT1 gene alterations in Korean patients with NK-AML.

Patients and Methods:
This study included a total of 75 patients with NK-AML. All patients received standard induction chemotherapy (‘7+3’ regimen) at the Asan Medical Center, Seoul, Korea between May 1999 and Dec 2006. WT1 exons 7 and 9 were amplified using polymerase chain reaction (PCR) and purified PCR fragments were directly sequenced. The clinico-laboratory data were retrieved from the AMC Leukemia Registry.

Results:
WT1 mutations were found in four patients (5.3%): two duplication mutations and two insertion mutations. All mutations were in exon 7 and none in exon 9. Two of four patients harboring WT1 mutation failed to achieve complete remission (CR) and died at 3.9 and 7.0 months after diagnosis of AML. One of two patients attaining CR was alive without relapse at 44.0 month and another died in CR at 4.7 months. WT1 SNP rs16754 was AA genotype (WT1AA) in 5 (3.7%), AG (WT1AG) in 29 (38.2%), and GG (WT1GG) in 41 (54.7%). The findings are significantly different from those of German report (WT1AA in 74.3%, WT1AG in 24.1%, and WT1GG in 1.6%) (P<0.0001). No significant difference of baseline clinico-laboratory findings was observed between different genotypes of WT1 SNP rs16754 (WT1AA/AG vs. WT1GG). The CR rates were similar between two SNP groups (WT1AA/AG vs. WT1GG, 85.3% vs. 80.5%, P=0.78). The five-year probabilities of overall (OS) and relapse-free (RFS) survival appeared to be lower in patients with WT1GG genotype, but the differences were not statistically significant (WT1AA/AG vs. WT1GG; OS, 49.3% vs. 36.9%, P=0.43; RFS, 56.3% vs. 42.5%, P=0.64).

Conclusion:
The prognostic impact of WT1 mutation in NK-AML could not be assessed in this study due to low number of patients harboring the mutation, but three of four patients died with relatively short survival duration, suggesting poor prognosis of the patients with WT1 mutation. The frequency of WT1 SNPs rs16754 in our patients was different from that of German report and survivals were not significantly different according to the SNP genotypes.

Disclosures:
No relevant conflicts of interest to declare.
The Activator of Transcription 3 (STAT3) Gene Polymorphisms Are Associated with Treatment Outcomes In Acute Myeloid Leukemia
Authors: Chen Bao-An , Zhong Yue-jiao , Feng Ji-feng , Cheng Lu , Li Yufeng , Qian Jun , Cheng Jian , Bao Wen , Ding Jia-hua 
Published: October 2019

Abstract
Abstract 4858

Objectives:
In the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway signaling pathway, the STAT3 is one of the most prominent predictable factors for cancer and leukemia. STAT3 activation might promote cellular transformation and therefore have an important role in human tumors. This study was aimed to investigate the relationship between the STAT3 polymorphisms and the treatment responses of AML in the Chinese population.

Methods:
We tested three single nucleotide polymorphisms (SNPs) with 130 Acute Myeloid Leukemia (AML) patients. Genomic DNA was isolated from peripheral blood and assayed by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF).

Results:
The results showed there were strong relationship between unfavorable cytogenetic, partial remission (even no remission) and the frequencies of GG genotype in rs9909659 (P=0.01 and 0.03) and the patients less than 45 years old was significant association with GA/AA genotype (P=0.01). But associations were not confirmed about event-free survival and leukemia-free survival in the Chinese population studied.

Conclusions:
It is clear that the AML patients with GG genotype in rs9909659 are not sensitive to standard chemotherapy method

Disclosures:
No relevant conflicts of interest to declare.
Polymorphisms In Signal Transducer and Activator of Transcription 5 (STAT5) and Response to Cytarabine-Based Chemotherapy In Acute Myeloid Leukemia Patients
Authors: Chen Bao-An , Zhong Yue-jiao , Feng Ji-feng , Cheng Lu , Li Yu-feng , Qian Jun , Ding Jia-hua , Gao Feng , Xia Guo-hua , Lu Zuhong , Cheng Jian , Bao Wen 
Published: October 2019

Abstract
Abstract 4859

Objectives:
Signal transducer and activator of transcription 5 (STAT5) protein is one of the concernful part of STATs families. The constitutive STATs activation has been especially showed in transformation by fusion genes in leukemias.

Methods:
In this study, we aimed to investigate whether the genetic polymorphisms in the STAT5 gene are associated with the treatment outcomes of Ara-C-based chemotherapy regimens in Acute Myeloid Leukemia (AML) patients. 152 AML patients in a Chinese sample were enrolled in our study. Peripheral blood samples were analyzed by matrix assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF-MS).

Results:
The results showed that the frequencies of the T/T genotype in rs2293157 and rs1135669 were higher in unfavorable and intermediate group respectively (P=0.023 and P=0.033). The frequency of patients with T/T genotype of rs1135669 was significantly higher in complete remission (CR) group.

Conclusions:
We concluded that the T/T genotype of rs1135669 might be an important marker for therapeutic efficiency of Ara-C-based chemotherapy in AML patients, especially in Chinese population.

Disclosures:
No relevant conflicts of interest to declare.
Rituximab, Endoxan, Ganciclovir, Dexamethasone, Tocilizumab Sequential Treatment In Kaposi's Sarcoma-Associated Herpesvirus (KSHV) -Related Multicentric Castleman Disease. A Case Report with 8 Months Follow-up
Authors: Iuliano Francesco , Mingrone Tecla , Infusino Stefania , Perricelli Alessia , Pomillo Angelo , Luci Maria , Maio Massimo Di 
Published: October 2019

Abstract
Abstract 4872
Clinical history of Kaposi sarcoma-associated herpesvirus (KSHV),-related Multicentric Castleman Disease is characterized by a rapidly progressive and often fatal course. We report a case successfully treated sequentially with a course of 4 infusions of rituximab 375mg/m2 at weekly intervals, Endoxan, 750 mg/m2 as an intravenous infusion once weekly for 2 weeks, intravenous ganciclovir, 5 mg/kg twice daily for 2 weeks and then once daily, dexamethasone 20 mg on days 1–4, 9–12, 17–20 and anti-interleukin-6 receptor antibody (tocilizumab) administered intravenously at a dose of 8 mg/kg every 2 weeks, starting at day 32.
On December 12, 2009, a 54-year-old man came to the medicine department because of a 2-week history of progressive fatigue, wasting, high-grade fever (39°C), profuse sweating, and severe autoimmune hemolytic anemia (4.2 mg/dL). He had been diagnosed with asymptomatic MCD 6 months before admission. Generalized lymphadenopathy and hepato-splenomegaly were evident on physical examination and enlarged lymph nodes in retroperitoneal regions (CT scan). Blood biochemistry showed very high CRP, IL-6, beta 2 microglobulin serum levels, low LDH and albumin, increase polyclonal immunoglobulin, grade 3 thrombocytopenia and neutropenia. Serologic tests (Epstein-Barr virus, hepatitis B and C viruses, cytomegalovirus and human immunodeficiency virus) were negative. On hospital day 10, 250.000 copies of HHV-8 DNA were detected in 1 mL plasma by RT-PCR. The patient achieved CR two months after admission, still maintained at 8 month follow-up. HHV-8 DNA became undetectable over the course of 8 weeks, at which time ganciclovir was discontinued.
Conclusions: In our case the onset of the disease was serious and life-threatening.
Sequential therapy has proven to be able to save the life of the patient acting on different biological targets. To date is the first case treated with this schedule.

Disclosures:
No relevant conflicts of interest to declare.
Comparing the Outcomes of CHOP Chemotherapy Alone with Rituximab Plus CHOP for Hong Kong Chinese Patients with Diffuse Large B-Cell lymphoma
Authors: Law Man Fai , Yip Sze-Fai , Chan Hay Nun , Yeung Yiu Ming , Choi Wai 
Published: October 2019

Abstract
Abstract 4894

Introduction
Diffuse large B-cell lymphoma (DLBL) is the commonest type of lymphoma worldwide. The condition is the same in Hong Kong and it accounts for about 30–40% of lymphoma cases. CHOP (cyclophosphamide, doxorubicin, vincristine and prednisolone) was used for treatment of DLBL for many years. Rituximab, a chimeric anti-CD20 monoclonal antibody was used for the treatment of DLBL for about 10 years. Previous studies have shown the better efficacy of rituximab plus CHOP over CHOP alone, including the remission rate, overall and event-free survival. No previous study compared the efficacy of rituximab plus CHOP with CHOP alone in Hong Kong Chinese.
We conducted a study to compare the outcomes of CHOP chemotherapy alone with rituximab plus CHOP in a local hospital in Hong Kong.

Method
It was a retrospective study from January 1999 to December 2009. Hong Kong Chinese patients with newly diagnosed diffuse large B-cell lymphoma were recruited in the study. They were divided into two groups. One group of patients was given CHOP chemotherapy and the other group was given rituximab plus CHOP. The remission rate, overall survival and event-free survival were compared in the two groups.

Results
62 patients were recruited in the study. Twenty-nine patients with median age of 55 (range 23–77) were given CHOP chemotherapy alone. Thirty-two patients with a median age of 53 (range 20–75) were given rituximab plus CHOP. The baseline characteristics were similar in both groups. There were 41% of male patients in each group. IPI (international prognostic index) score as well as the proportion of patients at stage III and IV disease were similar in both groups.
The complete remission rate was 38% in CHOP alone group and 78% in rituximab plus CHOP group (p=0.002, Fisher's exact test).
The 3-year overall survival rate was 47% in CHOP group and 74% in rituximab plus CHOP group (p=0.043). The 3-year event-free survival was also better in the rituximab plus CHOP group (p=0.026). 19 events (including relapse, progression and death) were observed in the CHOP alone group and 8 events were observed in the rituximab plus chemotherapy group.
The side effects profile was similar in both groups and the rate of infection is comparable in both arms.

Conclusion
This study has shown that the use of rituximab plus CHOP was better than CHOP alone in Hong Kong Chinese patients with newly diagnosed diffuse large B-cell lymphoma. The complete remission rate, overall survival and event-free survival were significantly higher in the rituximab plus chemotherapy group.

Disclosures:
No relevant conflicts of interest to declare.