bluuebunny/crossref_metadata_2025 huggingface dataset:
CCR2 Mediates Hematopoietic Stem and Progenitor Cell Trafficking to Inflammation Sites to Enhance Organ Repair.
Authors: Si Yue , Charo Israel , Tsou Chia-Lin
Published: October 2019

Abstract 2547

Poster Board II-524

Hematopoietic stem cells (HSCs) are bone-marrow derived, self-renewing pluripotent cells that give rise to terminally differentiated circulating blood cells. Recent work has implicated HSCs in the repair of parenchymal tissue in the setting of inflammation. In response to the antagonist of the chemokine receptor CXCR4, HSCs and their progenitors migrate from bone marrow to the blood. However, little is known about the signals that mediate their trafficking from the blood into peripheral tissues. Recently, we showed that mice genetically deficient in chemokine receptor CCR2 (CCR2–/– mice) have a marked decrease in the number of circulating “inflammatory” (7/4+, Ly6c+) monocytes, but no decrease in myeloid progenitor cells in the bone marrow (Tsou et al, J Clin Invest, 2007, 902). These data indicated that although CCR2 is not necessary for HSCs to differentiate into mature monocytes, it does play a role in monocyte egress from bone marrow to blood. In the current study, we extend this work and investigate the expression of CCR2 on hematopoietic stem cells and progenitor cells (HSPCs), and tested the hypothesis that CCR2 is critical for the recruitment of circulating HSPCs to sites of inflammation. Here we report that the chemokine receptor CCR2 is expressed on subsets of primitive hematopoietic stem cells as well as some early myeloid progenitors but not on the progenitors dedicated to megakaryocyte and erythroid differentiation. CCR2 mediates the chemotaxis of c-Kit+Lin- bone marrow derived cells to MCP-1 (CCL2) and MCP-3 (CCL7), which are known CCR2 ligands. In unchallenged mice, CCR2 appears to play a minimal role in HSCs trafficking. However, following instillation of thioglycollate to cause aseptic inflammation HSPCs were actively recruited to the peritoneum, as demonstrated by both FACS and functional colony formation assays. That these cells were true HSCs was demonstrated by their engraftment into the bone marrow of irradiated recipients. In contrast, mice genetically deficient in CCR2 (CCR2−/−) were profoundly impaired in the recruitment of HSCs to the inflamed peritoneum. Furthermore, in human disease models of acetaminophen induced liver damages, hematopoietic stem and progenitors were increased dramatically at site of injury in WT mice but not in CCR2−/− mice. CCR2 recruited HSPC play a tissue protective role by increase the numbers of M2 macrophages in injured liver. Taken together, these findings document a novel role for CCR2 in the specific homing of primitive hematopoietic stem and progenitors to sites of inflammation and injury, and suggest a new role for chemokines in increasing the numbers of differentiated leukocytes at sites of inflammation. Recruited HSPCs hasten the resolution of the inflammatory response, and promote repair of injured tissue, at least partially due to locally increase the number of tissue reparative macrophages.

Disclosures:

No relevant conflicts of interest to declare.
Pathway Based Evaluation of Cytarabine Pharmacogenetics in Children with Acute Myeloid Leukemia.
Authors: Myers Kasiani C. , Gerbing Robert B. , Alonzo Todd A. , Phillips Christine L , Gamis Alan S. , Radloff Gretchen A. , Perentesis John , Davies Stella M.
Published: October 2019

Abstract 2610

Poster Board II-586

Cytarabine is a key chemotherapeutic agent in the treatment of acute myeloid leukemia (AML). We have taken a pathway based candidate gene approach to select variants likely to be critical to cytarabine metabolism. Single non-synonymous nucleotide polymorphisms and polymorphisms located in promoter regions of genes involved in cytarabine metabolism, with known frequencies > 5% in the Caucasian population were identified. We genotyped 479 children treated for de novo AML on the Children's Cancer Group (CCG) 2941 and 2961 protocols for these polymorphisms. All patients received intensively timed induction therapy with IDA-DCTER (idarubicin, dexamethasone, Ara-C (800 mg/m2 total dose), thioguanine, etoposide and daunomycin) given on days 0 to 3 followed by DCTER given on days 10 to 13. On recovery of ANC and platelet counts, patients were randomized to consolidation therapy with Regimen A: a further course of IDA-DCTER/DCTER or Regimen B: IDA-FLAG (idarubicin, fludarabine, Ara-C (7590 mg/m2 total dose Ara-C) and G-CSF). Patients with a matched family donor received intensification with allogeneic transplant; all others received a single course of high dose Ara-C (24,000 mg/m2 total dose) and L-asparaginase. There were no significant associations between genotypes and clinical characteristics of leukemia, although polymorphism frequencies did differ significantly by race. Significant associations with outcome of therapy in a multivariate analysis adjusted for race are listed below:

The nucleoside transporter gene hENT promoter polymorphism rs747199 influenced 5 year overall survival (OS) from study entry (46 ± 6% in G/G cases versus 58 ± 9% in other patients (p=0.039). Similarly, 5 year OS from end of two courses was improved in children with a C/C genotype at rs693955 (OS 66 ± 7% vs. 54 ± 11%, p= 0.058). No significant differences were found in disease free survival, relapse rate (RR), or treatment related mortality for these variants. Nucleoside transporter polymorphisms may play a critical role in modulating cytarabine transport and influence the outcome of therapy for AML in children.

Disclosures:

No relevant conflicts of interest to declare.
Target Epitopes of Human T Lymphotropic Virus 1 Recognized by Class I MHC-Restricted Cytotoxic T Lymphocytes in HAM/TSP Patients and Infected Patients with Autoimmune Disorders.
Authors: Kozako Tomohiro , Akimoto Masaki , White Yohann , Toji Shingo , Matsushita Kakushi , Kubota Ryuji , Izumo Shuji , Suzuki Shinsuke , Uozumi Kimiharu , Shimeno Hiroshi , Soeda Shinji , Arima Naomichi
Published: October 2019

Abstract 2660

Poster Board II-636

Human T-cell leukemia virus-1 (HTLV-1) causes adult T cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) after long-term infection. A variety of collagen diseases such as Sjögren's syndrome and systemic lupus erythematosus has been reported in HTLV-1-infected individuals, although the precise relationship between these disorders and HTLV-1 infection remains unknown. We have previously reported the decreased frequency and function of HTLV-1 Tax-specific cytotoxic T lymphocytes (CTLs) in ATL patients due to insufficient cytolytic effector molecules compared with asymptomatic carriers (ACs) (ASH annual meeting 2007). However, there is no report on the repertoire of HTLV-1-specific CTLs in HAM/TSP patients or carriers with collagen diseases (CCs), both of which are characterized by abnormal immune states. In order to characterize HTLV-1-specific CTLs in ACs, HAM/TSP patients and CCs, we examined the frequency and diversity of HTLV-1-specific CTLs using 16 distinct HTLV-1 Tax and Env HLA-A*0201 and HLA-A*2402 tetramers. There was no statistically significant difference in the proportion of patients having HTLV-1 Tax specific CTLs among ACs, HAM/TSP and CC patients, but a greater proportion of patients with HAM/TSP or collagen disease had HTLV-1 Env-specific CTLs compared with ACs (Env175-183, Env239-246, Env442-450 for HLA-A*0201 and Env11-19, Env21-29, Env153-161 for HLA-A*2402) (Table 1). Within CD8+ lymphocyte subsets, the percentage of cells binding either Tax11-19/HLA-A*0201 or Tax 301-309/HLA-A*2402 tetramer (Figure 1), was significantly higher for HAM/TSP patients compared to ACs, being lowest among CCs. Additionally, the number of epitope repertoires found on HTLV-1 Env-specific CD8+ cells in ACs was considerably lower than for HAM/TSP patients and CCs (ACs, 1/6; HAM/TSP, 4/6; CCs, 6/6). This study demonstrates the relatively greater importance of CTLs recognizing HTLV-1 envelope tetramers compared to HTLV-1 Tax tetramers, thereby suggesting that the diversity, frequency and repertoire of HTLV-1-specific CTL clones, especially Env-CTLs, may be related to the hyperimmune response in HAM/TSP and carriers with collagen diseases.

Disclosures:

No relevant conflicts of interest to declare.
Interim Report of Phase II Study of Bortezomib Plus CHOP Every 2 Weeks in Patients with Disseminated Stage Diffuse Large B-Cell Lymphoma.
Authors: Kim Jeong Eun , Lee Dae Ho , Lee Soon Il , Lee Jae Hoon , Kim Won Seog , Yoon Bora , Suh Cheolwon
Published: October 2019

Abstract 2688

Poster Board II-664

Rituximab with cyclophosphamide, doxorubicin, vincristine, and prednisolone (R-CHOP) and dose-dense CHOP therapy has improved the treatment results of diffuse Large B-cell lymphoma (DLBCL) patients. Nevertheless, a substantial number of patients progress or relapse. Bortezomib is a proteasome inhibitor that is widely used in myeloma treatment and was recently approved for use in mantle cell lymphoma treatment. Its antitumor activity in DLBCL patients was reported in several studies, both as a single agent and in combination with chemotherapy. In this study, we tried to add bortezomib to dose-dense CHOP every 2 weeks to evaluate the complete response (CR) rate and safety of the treatment. Patients with previously untreated disseminated stage DLBCL and age 70 years or less were eligible. All patients received CHOP (Cyclophosphamide 750 mg/m2 IV, doxorubicin 50mg/m2 IV, vincristine 1.4mg/m2 IV on day 1, and prednisolone 100mg PO on days 1 to 5) and granulocyte colony-stimulating factor at a dose of 5μg/kg from days 4 to 13 every 2 weeks. Bortezomib, as recommended by the previous phase I trial, was administered at a dose of 1.6mg/m2 on days 1 and 4 of each cycle. Thirty-five patients were enrolled from March, 2007 to March 2009. Total 188 cycles of treatment were done. Twenty-six patients finished planned 6 cycles of treatment, while 9 patients could not continue all planned treatment due to treatment related toxicities and one patient experienced disease progression after 5 cycles of treatment. Twenty-four patients had a CR (92.4%), one patient had a partial response (3.8%), and one patient had a progressive disease (3.8%). Grade 3 hematologic toxicity episodes occurred including 20 anemia (10.7%), 4 neutropenia (2.1%), and 6 thrombocytopenia (3.2%). Grade 4 hematologic toxicity episodes comprised 2 anemia (1.6%), 11 neutropenia (5.9%), and 6 thrombocytopenia (3.2%). One patient died of severe infection with neutropenia. Among 35 patients, grade 3 non-hematologic toxicity occurred with fatigue (11.4%), nausea (2.9%), vomiting (5.7%), diarrhea (8.6%), abdominal pain (8.6%), and sensory neuropathy (20.0%). One patient experienced grade 4 constipation. This interim analysis shows that bortezomib plus dose-dense CHOP every two weeks showed promising activity in disseminated DLBCL patients as the first-line treatment with acceptable toxicity. Further accrual will be continued till the planned patient enrollment goal for phase II results.

Disclosures:

No relevant conflicts of interest to declare.
Long-Term Outcomes of Patients with Newly Diagnosed NK/T-Cell Lymphoma Treated by EPOCH Regimen.
Authors: Huang Huiqiang , Lin Zexiao , Lin Xubing , Cai Qingqing , Xia Zhongjun , Jiang Wenqi
Published: October 2019

Abstract 2689

Poster Board II-665

Introduction:

Extranodal NK/T-cell lymphoma (ENKL) is a distinct clinicopathological entity of non-Hodgkin's lymphoma, which is characterized by a highly aggressive clinical process and poor prognosis. There are no standard treatment strategies available now. The present study aimed to evaluate the treatment outcome and toxicity of EPOCH regimen for ENKL.

Methods:

From June 2003 to June 2008, 34 patients including 30 nasal ENKL (88.2%) and 4 extranasal ENKL (11.8%) received doxorubicin, vincristine, etoposide over 96 hours infusion with bolus cyclophosphamide and oral predinisone (EPOCH) combination as first-line treatment. By the Ann Arbor lymphoma Staging Classification, 29 patients (85.3%) had stage ±E-IIE disease and 5 patients (14.7%) had stage IIIE-‡WE disease. Median cycles of EPOCH regimen administered were 2.5 (range from 1 to 6 cycles). Additional involved field radiation therapy (IFRT) was administered to patients with localized regional disease after chemotherapy.

Results:

Among 34 patients, 33 were eligible for objective response evaluation. The response rate (RR) for the whole group was 60.6% (20/33) with complete remission (CR) rate of 45.5% (15/33). The RR of patients with nasal ENKL was 66.7% (20/30), the CR rate was 50% (15/30). Of the 4 patients with extranasal ENKL, only one achieved stable disease (SD), the other three had progressive disease (PD) during chemotherapy. After a median follow-up of 22 (2–68) months, the estimated overall 3-year overall survival rate (OS) was 52.2%. For patients with nasal ENKL, the estimated median survival time was not reached, the 3-year OS was 59.4%. For patients with extranasal ENKL, the estimated median survival time was only 7 months. For stage ±E-IIE patients with nasal ENKL who received initial EPOCH chemotherapy followed IFRT, the CR rate was 75.0%, and the 3-year survival rate was 75.0%. 102 cycles of chemotherapy were administered to 34 patients. Major adverse effect was myelosuppression. The incidence of grade III-‡W neutropenia was 30.9% and 60 cycles needed G-CSF support. Other toxicities were mild, no treatment-related mortality occurred.

Conclusion:

EPOCH regiment was effective on ENKL with acceptable toxlicity. EPOCH followed by IFRT yielded promising outcome for patients with localized nasal ENKL. However, patients with extranasal ENKL responded poorly, and more efficacious treatment strategies are urgently needed.

Disclosures:

No relevant conflicts of interest to declare.
Alterations of DCC Gene in B-Cell Malignancies.
Authors: Nagoshi Hisao , Taki Tomohiko , Nishida Kazuhiro , Goto Minako , Kobayashi Satoru , Tsutsumi Yasuhiko , Matsumoto Yosuke , Kuroda Junya , Horiike Shigeo , Otsuki Takemi , Taniwaki Masafumi
Published: October 2019

Abstract 4428

Many genetic alterations have been detected in various tumors to date, and these are concerned with oncogenesis closely. Oligonucleotide array analysis has contributed to detect genomic gains and losses implicated in the development of human malignant tumors, identifying either oncogenes or tumor suppressor genes. In our recent data, oligonucleotide array analysis showed recurrent genomic copy number variations at 18q21-22 in malignant lymphoma and multiple myeloma cell lines. There are a lot of genes related to oncogenesis at 18q21-22, such as SMAD4, DCC, MBD2, TCF4, TXNL1, WDR7, FECH, NARS, MALT1, and BCL2. We studied expression of these genes by reverse-transcriptase polymerase chain reaction (RT-PCR). Although most genes examined expressed in almost all cell lines examined, DCC (deleted in colorectal cancer) could be detected in a small number of malignant lymphoma cell lines.

DCC, which encodes a receptor for netrin-1, would control apoptosis and be concerned in oncogenesis. Loss of one DCC allele or decreased DCC expression occur in many solid tumors, suggesting that DCC inactivation constitutes a critical event in the development of these tumors. Similarly, the inactivation of DCC can have a role in the development of hematologic malignancies, including leukemia and malignant lymphoma. We analyzed DCC expression in 15 multiple myeloma and 9 malignant lymphoma cell lines, by RT-PCR using various primer pairs. However, no products could be detected using any primer pairs containing DCC exon 1 primer in three multiple myeloma cell lines, although other primer pairs could amplify products, suggesting the DCC transcripts lacking exon 1 in these cell lines. This result made us predict the abnormal form of DCC. Therefore, we tried to clone the DCC transcripts fused to unknown sequences instead of exon 1, using bubble PCR method for cDNA in these three cell lines. Some products were obtained and sequenceanalysis revealed that the bubble PCR products contained exon 2 of DCC fused to unknown sequences. BLAST search revealed that these unknown sequences were part of intron 1 of DCC, resulting in the lack of start codon in exon 1. Putative abnormal DCC protein lacks a part of N-terminus, suggesting that structural abnormalities of DCC play some roles in oncogenesis of hematologic malignancies. Although the previous studies described the abnormalities of DCC derived from allelic loss or decreased expression, structural abnormalities of DCC we found in this study might be another critical mechanism for DCC abnormalities even if the normal expression could be detected by RT-PCR which could amplify only limited portion of the gene.

Disclosures:

No relevant conflicts of interest to declare.
Research of Gene Expression in Patients with Myelodysplastic Syndromes.
Authors: Chen Bao-An , Zhang Bo , Gao Chong , Xia Guo-Hua , Shao Ze-ye , Ding Jia-Hua , Chen Jian , Wang Jun , Song Hui-Hui , Bao Wen , Zhong Yue-Jiao , Pei Xiao-Ping , Wang Fei , Chen Ning-Na
Published: October 2019

Abstract 4433

Object

This study was aimed to investigate the expression of c-FLIPL, c-FLIPS and DLK1 mRNA in Myelodysplastic Syndromes (MDS) patients, as compared with normal people and AML patients, and to find its clinical significance.

Methods

The expression of c-FLIPL, c-FLIPS and DLK1 mRNA in bone marrow mononuclear cells (BMNNC) of 16 patients with MDS, 8 patients with AML and 3 controls were detected by RT-PCR.

Results

The expression of DLK1 mRNA was up-regulated in MDS, including RA and RAEB, as compared with controls(P<0.05). There was no significant difference in expression of DLK1 between RA and RAEB(P>0.05). The expression of DLK1 was significant higher in AML patients, compared with controls(P<0.05). There was no significant difference between MDS and AML patients(P>0.05). The expression of c-FLIPL mRNA was higher than that in controls, both in RA and RAEB(P<0.05). There was no significant difference in expression of c-FLIPL between RA and RAEB(P>0.05). In eight AML patients, c-FLIPL gene's expression was up-regulated, as compared with controls(P<0.05). Between AML and MDS patients, there was no significant difference(P>0.05); The expression of c-FLIPS mRNA had no significant difference between MDS patients and controls(P>0.05), but its expression in RAEB was significant higher as compared with RA patients and controls(P<0.05). And in AML patients, the expression of c-FLIPS was higher than that in controls(P<0.05), but there was no significant difference between AML and MDS patients(P>0.05).

Conclusion

It is concluded that the expressions of DLK1, c-FLIPL and c-FLIPS mRNA in MDS/AML patients are abnormal as compared with normal people, although there are no significant difference have been found between AML and MDS. These genes may play critical roles in escaping malignant clone of MDS from apoptosis and acquiring the ability to divide unlimitedly, they can become important indexes for evaluating of development in MDS.

Disclosures:

No relevant conflicts of interest to declare.
Impact of Therapeutics On Fatigue in Chronic Immune Thrombocytopenia (ITP).
Authors: Bussel James B.
Published: October 2019

Abstract 4455

Complaints of tiredness, lethargy or lack of energy are common in patients with chronic Immune ThrombocytoPenia (ITP), however they are not well understood. This study assessed whether different clinical and laboratory aspects are related to fatigue and how different therapeutic modalities might impact these symptoms.

Methods

Patients with chronic ITP older than 15 years old were enrolled from 2007-2008 at the Platelet Disorders Center at the Weill-Cornell Medical Center, NY. They completed the Short-Form 36 questionnaire (SF-36) and Fatigue severity scale (FSS) (2 standardized Health-related quality of life [HRQoL] questionnaires) and the Beck Depression Inventory (BDI). Laboratory testing included CBC, TSH, ferritin, iron + TIBC, and blood serotonin levels.

Results

83 patients with a mean age of 40.2 years (67.5% women) were enrolled. The mean platelet counts were 109,000/ul (range of 12,000-500,000/ul). Forty-one % of patients had duration of ITP of 10 years or more and 25% had had previous splenectomy. Sixty-one% were on treatment: 11% steroids, 23% Tpo-R agents and 28% “Other” (7.2 % Rigel, 13.3 % IVIG, 3.6 % anti-D, 2.4 % rituximab and 1.2% Danazol/Azathioprine). Surprisingly, analysis of the SF36 showed that the study (ITP) population is similar to the general US population and that they do not report increased fatigue or depression. These results contrast with a previous report by McMillan, AJH, 2007 (fig 1). However when the current patients were divided according to age, abnormal low scores on SF36 were observed on vitality domain (mean 50) for the age group of 45-55 years and on role physical domain (mean 48) for the age group of 55-65 years. Furthermore, differences were observed in patients stratified according to treatment (figure 2) where the mean scores for patients on the steroid group on the respective domains were lower: vitality 49 and role physical 42. The Tpo-R agonists group had better results in all SF36 domains; similar findings were seen with the FSS and BDI. The only laboratory finding is that patients with low ferritin levels had worse results on FSS.

Conclusion

Although fatigue and related symptoms have been previously reported by others including an improvement with successful treatment eg in studies of the TPO-R agonists, to our surprise we did not find the same results. Whether this study population is intrinsically different or receiving different treatments, eg avoidance of steroids, might impact these results here are 2 possible hypotheses. Use of different fatigue assessments as ITP-PAQ or CDC symptom inventory and further studies addressing the role of the pro-inflammatory cytokines in these patients would be useful.

Disclosures:

No relevant conflicts of interest to declare.
TNF-α and Its Receptors in Patients with Acute Myeloid Leukemia Undergoing alloHSCT.
Authors: Kyrcz-Krzemien Slawomira , Zielinska Monika , Wieclawek Agnieszka , Czerw Tomasz , Holowiecka Aleksandra , Zielinska Patrycja , Mazurek Urszula
Published: October 2019

Abstract 4484

Introduction

Several studies have demonstrated, that TNF α and its receptors (sTNFR) can be used as prognostic markers for major transplant related complications (TRC) after allogeneic haematopoietic stem cell transplantation (alloHSCT). Measurement of that inflammatory cytokine and sTNFR during pretransplant conditioning and early after transplantation may also reflect the conditioning-induced tissue damage.

Patients and methods

Our study included a group of 36 adult patients with acute myeloid leukemia (AML) in complete remission, who underwent alloHSCT following standard myeloablative conditioning according to Bu/Cy protocol (17 patients) or reduced-toxicity myeloablative conditioning with treosulfan-based regimens (19 patients). 21 patients received alloHSCT from a sibling and 15 from an unrelated donor. The expression of TNF-α, TNFRI and TNFRII were analyzed using reverse transcription-polymerase chain reaction (RT-PCR) in peripheral blood mononuclear cells (PBMC) before the start of conditioning, on the day of transplantation and on day 30 after alloHSCT. We examined β-actin as well, which constitutes an endogenous amplification control and is a housekeeping gene. sTNFRI and sTNFRII serum levels were measured using an enzyme-linked immunosorbent assay (ELISA) at the same defined time points before and after alloHSCT. The Mann-Whitney U test was used to evaluate the significance of differences between the analyzed groups and the Shapiro-Wilk and Lilliefords tests to estimate data distribution.

Results

TNFRII mRNA was not detected in PBMC, but TNF-α and TNFRI mRNA as well as β-actin were discovered. The real time PCR showed significant decrease (p<0.004) in TNFRI expression on day 30 after transplantation in comparison to day 0. On day 30 expression of TNFRI was higher in patients treated with Treosulfan-based regimens than in patients treated with Bu/Cy (p=0.027). Plasma levels of sTNFRI and sTNFRII observed on day 0 and 30 were significantly elevated (p<0.05) compared with the levels prior to conditioning.

Conclusion

Our data indicate that sTNFRI and sTNFRII can be investigated as the markers reflecting the toxicity of the conditioning regimens in AML patients undergoing alloHSCT. The associations between TNF-α, TNFRI expression, serum levels of sTNFR and the clinical outcome after alloHSCT should be evaluated.

Disclosures:

No relevant conflicts of interest to declare.
Targeted Therapy by Using Tissue Specific Promoter and Granules as Storage Compartments for Therapeutic Proteins in Hematopoietic Cells.
Authors: Afsharian Parvaneh , Johansson Åsa , Persson Ann-Maj , Olsson Inge , Hansson Markus
Published: October 2019

Abstract 4508

Systemic administration of immunotherapeutics often give rise to severe side effects. A local deposition, using secretory lysosomes of hematopoietic cells as vehicles for delivery, may overcome this problem. Our research regarding sorting mechanisms of granule proteins have led to a new therapeutic idea – targeted therapy with hematopoietic cells as carriers. Exogenous proteins are expressed, and the gene product is directed for secretory lysosome or granule storage. These organelles are unique storage compartments in neutrophils for biological active proteins, designated for inflammatory foci. The exogenous proteins, would accordingly, be locally released at the site of inflammation. Our model construct, soluble TNF receptor 1 (sTNFR), can be directed for storage by adding a transmembranous region and a sorting signal. We have earlier shown that this construct, retrovirally expressed in murine hematopoietic progenitor cells, are destined for granule targeting. Furthermore, we have transplanted transduced murine progenitor cells with modified granule content, and found stable expression and inducible release of sTNFR1 in vivo. Our goal is cell linage specific expression in order to selectively express sTNFR1 with a transmembranous region and a signal peptide for granule targeting. To achieve this, lactoferrin, perforin and MPO promoters, including regulatory elements, have been inserted into vectors (we use a retroviral vector with self-inactivating LTRs). The vectors is retrovirally transduced into murine hematopoietic progenitor cells, and the cells are cultured during cytokine driven differentiation and analyzed by FACS and Western blotting. Our results suggest that a concept of using secretory organelles of hematopoietic cells as vehicles for therapeutic agents in vivo may become feasible. It will be possible to test this principle by experiments in animal models of inflammatory and malignant diseases. Likewise, development of this concept may give new information on secretory lysosome function and secretion at inflamed/malignant sites.

Disclosures:

No relevant conflicts of interest to declare.
Five Pro-Inflammatory Molecules Polymorphisms Analysis in Ischemic Cerebrovascular Disease (ICD).
Authors: Moreno Vanessa P. , García-Raso Aránzazu , García-Bueno María José , García Araceli , Sánchez Carmen , Subirá Dolores , LLamas Pilar
Published: October 2019

Abstract 4514

Introduction

Ischemic cerebrovascular disease (ICD) is a multifactorial entity whose anatomopathological substrate is the pro-inflammatory state links to atherosclerosis. The inflammatory cells migrate to vascular endothelium unleashing atheroma plaque formation and pro-inflammatory cytoquine release.

Objective

The purpose of this case-control study is to analyze the ICD risk associated to polymorphisms MCP-1 -2518 G/A, IL6 -174 G/C, IL 1B -511 C/T, IL1B +3954 C/T, IL1A -889 C/T.

Methods and patients

A total of 282 ICD diagnosed patients (134 female and 148 male; mean age 68,2±14,9 years) and 287 controls without thrombotic history (143 Female and 144 male; mean age 65,8±14 years) were analysed. AH, DM, dyslipidemia and smoking habit data were collected for each group. Polymorphisms were analysed by PCR-RFLP. Seric IL6 was measured by ELISA. Statistic Analysis was made with Stata v.10 software.

Results

Stroke subtype distribution was: atherothrombotic (32,8%), inhabitual and indeterminate (28,9%), cardioembolic (24,5%), and lacunar (13,7%). Risk factors were more frequent in patients with AH and DM showed a higher prevalence: (OR: 1.684 IC 95%:1.161- 2.443, p:0.006) and (OR: 2.393, IC95%:1.447- 3.956, p:0.01) respectively. Logistic regression was used to determinate polymorphisms distribution. Results are shown in table 1. Analysis of IL-1 polymorphisms showed a synergic relation in IL1A -889 C/T (p:0.05; OR: 0.625; IC95%:0.388-1.008). The G allele IL6 –174 G/C carriers group presented higher seric IL6 levels than C allele carriers, 5.85 pg/ml (2.9- 34.5) vs 4.94pg/ml (2.5- 11.4) respectively. IL-6 genotype frequency G/G + G/C vs C/C was analyzed. Atherothrombotic stroke IL-6 genotype frequency was G/G + G/C (24.3%) vs C/C (12.0%) (p: 0.082).

Conclusion

There is no association between MCP-1 -2518 G/A, IL6 -174 G/C, IL 1B -511 C/T, IL1B +3954 C/T, IL1A -889 C/T polymorphisms and ischemic stroke. We found a synergic effect between IL1 mutations. The G allele IL6 –174 G/C carriers group presented higher seric IL6 levels than C allele carriers. The analysis performed by ICD subtypes showed an association between IL6 G allele and atherothrombotic subtype.

* SETH and MMA GRANT

Disclosures:

No relevant conflicts of interest to declare.
Experience with Hemophagocytic Lymphohistiocytosis at a Single Institution.
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 4517

Background

Hemophagocytic lymphohistiocytosis/macrophage activation syndrome (HLH/MAS), whether primary or secondary, is a serious and potentially life- threatening histiocytic disorder and generally fatal without immune modulating chemotherapy. This study aimed to define the clinical spectrum and treatment outcome in Korean children.

Patients and Method

Patients who were diagnosed with HLH/MAS at Asan Medical Center between October 1997 and April 2009 were identified and charts were reviewed.

Results

Twenty-six patients (16 male, 10 female) were evaluated during 13 - year period. Median age at diagnosis was 29.5 months (range: 0 - 180 months). Twelve patients had HLH secondary to underlying disorders, including viral infection, autoimmune disease. Epstein-Barr virus (EBV) - associated HLH was present in 9 patients, 1 of whom died, while 4 of 17 non-EBV associated patients died. Initial treatment with intravenous immunoglobulin (IVIG) was given to 9 patients and they had no response to IVIG. Eventually, all the patients except two underwent full protocol therapy (HLH 1994/2004) and the 2 patients died of rapid disease progression with delayed diagnosis. Seven patients revealed disease reactivation, median 3 months from HLH protocol therapy, and further treatment using the same regimen induced second remission of short duration. Seven patients (26.9%) had evidence of central nervous system disease at the time of diagnosis and neurological symptoms consisted of seizure, irritability, vomiting. Five of them are long-term survivors including one who received allogeneic stem cell transplantation (SCT) and all 5 patients show normal neurological function after treatment. The familial form of HLH was identified in 3 patients and all received allogeneic SCT from unrelated donor in second remission. They are still alive without relapse. At a median follow-up of 17.5 months, the estimated one-year overall survival rate was 82.8 ± 7.9 %.

Conclusion

Treatment outcomes using HLH 1994/2004 protocols in Korean children have become comparable to those reported in an international collaborative study. And allogeneic SCT represents a curative treatment for familial form of HLH. Further studies are required to more precisely define clinical spectrum and treatment outcome.

Disclosures:

No relevant conflicts of interest to declare.
Hodgkin's Lymphoma in the Elderly Patient: Impact of Age and Co-Morbidities On Outcomes.
Authors: Nakkar Samer , Pacioles Toni , Ballester Gabriela , Tirona Maria , Ballester Oscar F
Published: October 2019

Abstract 4529

Introduction

Age older than 45 years is a recognized independent prognostic factor for patients with Hodgkin's Lymphoma (HL), and it is part of the International Prognostic Score (IPS) system. This score system was based on data derived from selected patients included in clinical trials. It is unclear from these studies if age affects primarily disease biology or the ability of older patients to tolerate therapy.

Patients and Methods

We retrospectively reviewed all consecutive patients with newly diagnosed HL at our institution. Data collected included their IPS and a co-morbidity score (CoM) which includes assessment of co-existing cardiac, hepatic, pulmonary, renal and other morbidities.

Results

Forty five patients were identified. Twenty nine patients (64%) were younger than 45 years and 16 (35%) were 45 years or older. Patients were treated wit ABVD (adriamycin, bleomycin, vinblastine and DTIC) chemotherapy, with or without radiotherapy, except for 4 patients who were accrued to clinical trials. An IPS of 2 or more was documented in 17% of younger patients as compared to 71% of those older. There was a significant association between age and IPS score (p= 0.001). Similarly, there was a significant association between age and CoM score (p= 0.01). A CoM score of 4 or higher was seen in only 7.6% of the younger population but in 46.6% of the older patients. With a median follow up of 62 months, overall survival is 86 % for the entire population. Overall survival for the younger patients is 93% and for the older 71% (p= 0.01). Five of the six documented deaths occurred in patients 45 years or older, and 4 of these 5 were seen in patients over 60 years of age. Crude death rates for patients <45 (n= 29), 45 to 59 (n= 9) and 60 or older (n= 7) are: 3.4%, 11% and 57% respectively.

Conclusions

Older patients are more likely to present with high IPS and CoM scores which explain at least in part their poor outcomes. A high mortality rate in patients >60 years underscores the need to explore new therapeutic approaches in the older population.

Disclosures:

No relevant conflicts of interest to declare.
Chemotherapy Induced Neutropenic Fever Seen at Emergency Room: A Review of 102 Cases.
Authors: Lim Joo Han , Yi Hyeon Gyu , Lee Moon Hee , Kim Hoon , Kim Chul Soo
Published: October 2019

Abstract 4530

Introduction

Febrile neutropenia (FN) is a major toxicity of chemotherapy in cancer patients requiring prompt medical measures. Although the FN is a situation of oncologic emergency, the diagnosis and treatment may vary among institutions, and how fast the patients are being treated often depends on competence of House Officers at Emergency Room (ER). There has been limited data regarding the clinical outcome of patients with FN who were brought to ER.

Patients and Methods

We evaluated the clinical manifestations, therapeutic outcomes, and risk factors of FN in a retrospective analysis of 102 adult patients who visited ER from January 1, 2006 to March 31, 2009. FN was defined as a body temperature >38°C and a neutrophil count >0.5 × 109/L on the day of fever or the day after.

Results

The ECOG performance status (PS) was 0 in 15 patients, 1 in 67, and 2 in 20. The patients had a mean age of 57 years (range, 25–84). Fifty four patients were male, 52 female. Underlying diagnosis was lymphoma in 30, and soid tumors other than lymphoma in 72. The mean ANC was 436.8/mm3 (range, 0–1000); 4 patients had an ANC of <10/mm3. Twenty-three patients (22.5%) died of complications related to FN. There was no statistical difference in therapeutic outcome between tumor types, i.e. lymphoma versus non-lymphoma (p=.521), PS (p=.438), sex (p=.099), depth of neutropenia (p=.162), or time intervals from visiting ER to starting antibiotic therapy (p=.414). Age was important prognostic factor in therapeutic outcome. The median age in fatal cases was 62 years where as that of non-fatal cases was 54 (p=.016). Bacteremia was documented in 19 patients among whom 10 (53%) died. Mortality was significantly higher in patients with blood culture proven bacteria than patients whose blood culture yielded no organism (p=.013).

Conclusion

Cases of FN seen at ER showed substantial mortality especially in elderly patients. Given the rising age in cancer diagnosis as well as therapeutic intervention, higher mortality rate associated with chemotherapy induced FN in elderly patients needs further study seeking the way to reduce the risk of death

Disclosures:

No relevant conflicts of interest to declare.
Jamshidi needle Biopsy of the Sternal Bone Marrow.
Authors: Davila Enrique
Published: October 2019

Abstract 4548

The aspiration and biopsy of the bone marrow is one of the most valuable and important tests in hematology, oncology and medicine. It is a high yield, safe, fast and informative test performed frequently in medical practice with minimal complications. For reasons of ease and safety, bone marrow aspiration and biopsy are usually obtained from the posterior iliac crest. I reviewed my experience in obtaining 37 consecutive bone marrow biopsies from the sternum using a Jamshidi needle (gauge 11; external diameter 3.048 mm) in 36 consecutive patients (twice in one patient) over a 9 year period, in whom a posterior iliac crest study could not be done.

Technique

After performing the sternal bone marrow aspiration in the usual manner, a small skin incision is made over the sternum with a scalpel. The Jamshidi needle is introduced at approximately a 90 degree angle in the middle of the sternum at the level of the 3rd intercostal space. After a ”give” is felt, indicating that the needle has reached the bone marrow cavity, the tip of the needle is angled downwards at 45 degrees or less and with a clockwise - counterclockwise movement, the needle is advanced for 3 to 10 mm. After a slight change of angle aiming at “breaking” the distal attachment of the bone marrow piece, the needle is slowly withdrawn with the same rotatory movements. In no case did I feel that I had reached the inner table of the sternum. All patients were observed and examined 20 minutes and 24 hours after the procedure.

Results

There were 22 inpatient and 15 outpatient procedures. The reasons that precluded the performance of the preferred posterior iliac crest bone marrow biopsy were: immobility in 17 patients, obesity in 13, prior radiation in 3 and other in 4. The final diagnosis was a malignant disorder in 17 patients (leukemia, lymphoma, myelodysplasia, plasma cell dyscrasia or metastatic cancer). All but one were new diagnoses. In 20 cases the final diagnosis was a benign hematological disorder or a non diagnostic bone marrow examination. In 9 occasions (mostly obese patients and patients with prior radiation therapy) a previous attempt at performing a posterior iliac crest biopsy had failed. The only complications were the development of a tumor nodule in the needle tract in one patient with an aggressive, Burkitt's type lymphoma and a small superficial hematoma in a patient with a highly vascular metastatic breast cancer.

The bone marrow core biopsy of the sternum, performed as described, in the hands of an experienced practitioner is a safe and helpful test in the evaluation of the bone marrow cytology, architecture and anatomy in selected patients in whom the performance of the preferred posterior iliac crest biopsy cannot be done.

Disclosures:

No relevant conflicts of interest to declare.
Implementation of Hematology-Oncology Lecture Series for Housestaff.
Authors: Steinberg Amir
Published: October 2019

Abstract 4554

BACKGROUND

Currently, the hospital has a floor dedicated to cancer patients. The housestaff taking care of these patients includes three interns and one hematology-oncology fellow. In addition, there is a consult service comprising another fellow, an intern, and a resident. There was a paucity of formal, didactic, teaching in place devoted to the subjects of hematology and oncology.

OBJECTIVE

the development of a monthly lecture series for housestaff rotating on a hematology-oncology rotation that would focus on three key areas:

METHODS

Topics were chosen based on pertinence. These included: leukemia, lymphoma, multiple myeloma, stem cell transplantation (these first four comprising at least 50% of patients on the unit), coagulation, breast cancer, lung cancer, colon cancer, gynecological malignancies, and pain management/supportive care. Potential speakers included full-time faculty as well as private physicians affiliated with the cancer center. Emails and/or phone calls were placed to the physicians and announcements made at division meetings and schedules developed. The plan was also for the development of a series of questions testing the housestaff at the start of their one month rotation and at the end of the rotation. Lectures were scheduled for afternoons if possible to avoid conflicts with morning rounds. Occasional morning talks were necessary to accommodate lecturers. Format of lectures was flexible (power point, dry-eraser board).

RESULTS

Overall feedback was positive for the lecture series. The two conditions affecting optimal success of the program concerned scheduling, including intern and fellow schedules as they relate to rounds and housestaff clinics and availability of when speakers could lecture and recruitment of speakers appropriate to the relevant topics. In addition, repeating the topics every month proved challenging for finding lecturing participants. The question/answer testing is still early in testing and no conclusion can be made.

CONCLUSION

A didactic lecture series is an important aspect of hematology-oncology education for housestaff. Repeating lectures every month is a challenge for several reasons. Possible interventions in the future may include:

Disclosures:

No relevant conflicts of interest to declare.
The Activation of STAT3 Contributes to the Effect of Autocrined G-CSF On the Proliferation of HL60 Cells Induced by ATRA.
Authors: Yang Weihua , Wen Peie , Liu Chuanfang
Published: October 2019

Abstract 4565

Background

In previous works, we detected several significant cytokines in the serum of patients with APL after treatment with ATRA, and the results revealed a close association of serum G-CSF and the number of promyelocytes or more mature granulocytes. Some of the results also indicate that ATRA in vitro has a conspicuous effect on the expression of some cytokines, such as IL-1β, IL-8, TNF, G-CSF, and so on. At least, it is showed that cytokines in serum or supernatant secreted by APL cells perhaps played an important role in the genesis of leukocytosis. However, the role of G-CSF in proliferation of APL cells has not yet been explained clearly. In the present study, we analyse the secretion and expression of cytokines in APL patients who has hyperleukocytosis, study the relationship between cytokines and leukocytosis, and detect the signal transduction pathway of cytokines.

Objective

To detect the signal pathway of IL-1β and G-CSF secreted by primary acute promyelocytic leukemia (APL) cells induced by all-trans retinoic acid (ATRA).

Methods

Primary APL cells from ATRA-treated patients were cultured in RPMI 1640, then the level of supernatant IL-1β and G-CSF were detected by ELISA assay. The expression of IL-1β or G-CSF mRNA was measured by RT-PCR, and the level of STAT1α and STAT3 evaluated by Western blot.

Results

Treated with ATRA, the peripheral blood leucocytes of patients with APL was obviously up-regulated, and peaked at 12 days averagely, of which mainly were myelocytes and metamyelocytes. The secretion of IL-1β or G-CSF in supernatant was up-regulated (P<0.05). RT-PCR assay confirmed that in the primary APL cells, with higher secretion of IL-1β and/or G-CSF, the expression of IL-1β and/or G-CSF was up-regulated, correspondingly. Cytokines and signal transduction factors analysis indicated that the expression of STAT1α protein increased in all the patients, irrelevant to the expression of IL-1β or G-CSF, but the increase of STAT3 expression was consistent with the expression and secretion of G-CSF.

Conclusion

The higher expression of G-CSF in primary APL cells induced by ATRA contributes to the activation of STAT3.

Disclosures:

No relevant conflicts of interest to declare.
Investigation of the Mechanistic Events Governing Platelet Release.
Authors: Thon Jonathan Noah , Anderson-Daniels Jordan M. , Braithwaite Matthew , Larson Mark K. , Hoffmeister Karin M. , Hartwig John H. , Italiano Joseph
Published: October 2019

Abstract 4590

Megakaryocytes generate platelets by remodeling their cytoplasm into long proplatelet extensions from which platelets release. While the mechanics of proplatelet elongation have been studied, the terminal steps of platelet production remain poorly understood. To understand the mechanics of platelet release, we developed a novel gradient sedimentation protocol to isolate released proplatelets. This has enabled us to define and quantify different stages in platelet maturation and identify a new intermediate stage in platelet production—the preplatelet. Preplatelets are anucleate discoid particles considerably larger (2-10μm) than platelets that have the capacity to reversibly convert into barbell-shaped proplatelets. Video-microscopy of GFP-tubulin expressing cells reveals that preplatelets elongate and twist into barbell forms by a process mediated by microtubule-based forces. α-Granules and dense granules present in the preplatelet translocate between platelet-sized swellings during barbell proplatelet formation, and ultimately concentrate at the terminal loops. Fission releases two mature platelets from the barbell ends, and isolated proplatelets will mature into bona fide platelets in culture over a 5-day period. Correspondingly, mature platelets are generated within 2 hours post-transfusion of fluorescently-tagged proplatelets into recipient mice. Released proplatelets were plated on extracellular matrix proteins (fibrinogen, laminin, fibronectin, collagen, matrigel) or a BSA control, and cultured for 24 hours to recapitulate the bone marrow vascular niche and determine whether these cell-substrate interactions mediate platelet release. Fibrinogen and laminin binding increased platelet release roughly 4-fold, suggesting cell-substrate interactions in the bone marrow vascular niche and surrounding vasculature are critical for the final stages or platelet production. These findings confirm that proplatelets mature into multiple individual platelets both in vivo/vitro, and begin to define the mechanistic events leading to platelet production.

Disclosures:

No relevant conflicts of interest to declare.