bluuebunny/crossref_metadata_2025 huggingface dataset:
5 Year Experience in Chronic Phase-Chronic Myeloid Leukemia Treated with Imatinib Copy Drugs in a Peruvian Hospital
Authors: Torres Aimee , Navarro Juan Ramon , Untama Jose , Moreno Mariela , Murillo Sergio
Published: October 2019

Abstract 4449

BACKGROUND AND OBJECTIVES:

In our Hospital patients with Chronic Myeloid Leukemia (CML) in chronic phase (CP) were treated with imatinib (Glivec) 400 mg qd from june 2001 to june 2006. Since july 2006, only imatinib copy drugs (brand names: Zeite, Imatec, Imatib, Millatus and Celonib, all of Indian origin) are being used to treat this disease at the Rebagliati Hospital in Lima Peru. We present here the efficacy, security, event-free survival, progression free survival, overall survival of patients treated with imatinib copy drugs.

METHODS

Retrospective and descriptive analysis of CP-CML patients treated only with imatinib since july 2006. They all initiated the treatment with 400 mg qd. Five year experience is presented here.

RESULTS

We analyzed data from 51 patients (pts) with CP-CML. Median age at diagnosis was 45.1 years (15–90), 49% female and 51% male, all patients were Chromosome Philadelphia positive. Low, Intermediate and High Hasford Score were 32%, 48% and 20%, respectively. Median time from diagnosis to start of imatinib was 7.6 months (0–123). Five patients (9.8%) began treatment more than six months after diagnosis. All patients had at least a 3 month cytogenetic monitoring.

Thirty five (68.6%) and 39 patients (76.4%) achieved Complete Cytogenetic Response (CCyR) and Mayor Cytogenetic Response (MCyR), respectively. Median time to CCyR was 7.6 months and 71.4% of these patients achieved it during the first six months of treatment. CCyR and MCyR were 38% and 67% at 3 months, 57% and 70% at 6 months, 63% and 83% at 12 monhts, 78% and 91% at 18 months, respectively. Quantitative RT-PCR was performed in 22 patients with CCyR and 18 pts (81.8%) had Mayor Molecular Response (MMR) in five years of follow up.

Four pts (7.8%) progressed to Blastic Phase and all of them had Low Hasford Risk at diagnosis. Eight pts (15.6%) changed to 2nd line treatment. Two pts (3.9%) abandoned treatment.

Only 1 pt (1.9%) had both anemias 3–4. Eighteen pts (35.2%) had neutropenia-3, 8 pts (15.6%) had neutropenia-4, 7 pts (13.7%) had thrombocytopenia-3 and 4 pts (7.8%) had thrombocytopenia-4. Neutropenias 3–4 were managed with filgrastim, as treatment and as prophylaxis.

At 5 years, Event Free Survival (EFS), Progression Free Survival (PFS) and Overall Survival (OS) were 82%, 89% and 91%, respectively (Figure 1, 2 and 3).

CONCLUSIONS

In our hospital, CP-CML patients treated with imatinib copy drugs obtained important cytogenetic and molecular responses with prolonged EFS, PFS and OS and an acceptable safety profile. Hasford Score was not a good prognosis predictor. Treatment with Imatinib copy drugs has proven to be an acceptable treatment in CML patients in our hospital.

Disclosures:

Torres: Bristol Myers Squibb: Speakers Bureau.
Frontline Autologous Stem Cell Transplantation As Intensive Consolidation in Patients with Peripheral T Cell Lymphomas: Multicenter Phase II Trial in Korea
Authors: Ahn Jae-Sook , Yang Deok-Hwan , Chae Yee Soo , Sohn Sang Kyun , Kwak Jae-Yong , Yhim Ho-Young , Lee Se-Ryeon , Jung Sung-Hoon , Bae Soo-Young , Kim Yeo-Kyeoung , Kim Hyeoung-Joon , Lee Je-Jung
Published: October 2019

Abstract 4477

Peripheral T cell lymphomas (PTCLs) are an aggressive subtype of non-Hodgkins lymphoma and they have shown the shorter survival compared with B cell lymphoma. High-dose chemotherapy (HDT) followed by autologous stem cell transplantation (HDT/ASCT) for PTCLs has a potential meaning of consolidating remission for PTCLs. However, the effectiveness of ASCT on distinct conditioning regimens, the optimal transplant timing in the frontline or relapsed are still unclear.

We investigated the clinical outcomes of HDT/ASCT as frontline intensification in 46 patients with newly diagnosed PTCLs except ALK(+) anaplastic large cell lymphoma. Patients underwent ASCT with a uniform conditioning regimen (busulfex, cyclophosphamide and etoposide). The median age was 47 years (17–65). The histological subtypes were 47.9% PTCL-NOS (n=23), 18.8% anaplastic large cell lymphoma (n=9), 4.2% angioimmunoblastic T cell lymphoma (n=2), 25% extranodal NK/T cell, nasal type (n=12), 2.1% hepatosplenic T cell lymphoma (n=1) and 2.1% enteropathy-associated T cell lymphoma. Thirty patients (62.6%) presented with advanced stage disease (III/IV) and 16 (33.3%) had B symptoms. At diagnosis, 21 patients (43.8%) were classified as high-intermediate/high risk by the age-adjusted IPI (aaIPI) and 10 (20.9%) were classified as high-risk (more than 2 factors) by the prognostic index for PTCL (PIT). Thirty-one patients (67%) could undergo HDT/HSCT and disease status at pretransplant consisted of 23 patients (50 %) with CR and 8 patients (17.4%) with PR. 6 out of 8 patients with PR at pretransplantation improved the response to CR after HDT/ASCT. There was no significant difference of the response rate between CHOP alone or CHOP-like chemotherapy and non-anthracycline-based chemotherapeutic regimen. At a median follow-up of 32.9 months, 23 patients (50%) are alive. The 5-year probability of overall and progression-free survival (PFS) was 48.2 ± 8.1 % and 47.4 ± 8.1%, respectively. However, the 5-year OS and PFS rate in transplanted patients was 57.3± 10.2 % and 55.3 ± 11.3 %, respectively.

Conclusion:

Frontline HDT/ASCT in patients with PTCL could be performed with a high response rates and a substantial impact on improving outcome for PFS. Our findings also indicate that busulfex, cyclophosphamide and etoposide is a feasible conditioning regimen in ASCT for PTCLs.

Disclosures:

No relevant conflicts of interest to declare.
NVP-BEZ235, A Dual Inhibitor of Phosphoinositol-3-Kinase (PI3K) and Mammalian Target of Rapamycin (mTOR), Is a Potent Inhibitor of Lymphoma Cell Growth and Survival
Authors: Buglio Daniela , Lemoine Manuela , Neelapu Sattva S. , Vega Francisco , Berry Donald , Younes Anas
Published: October 2019

Abstract 4965

The Phosphatidylinositol-3-kinase (PI3K)/AKT/mTOR pathway is frequently deregulated in Hodgkin (HL) and non-Hodgkin lymphoma (NHL), and has been linked with tumor cell growth and survival. Although several proteins/enzymes in this pathway can be targeted by a variety of small molecules in vitro and in vivo, it remains unclear which protein target is the ideal for clinical testing. Previous studies demonstrated that the clinical activity of mTOR inhibitors may be attenuated by a negative feedback loop that involves activation of AKT, suggesting that a dual inhibition of AKT and mTOR activation may produce a better therapeutic outcome. To test this hypothesis, we evaluated the in vitro activity of NVP-BEZ235, a dual inhibitor of PI3K and mTOR, in a panel of 13 HL and NHL cell lines. NVP-BEZ235 inhibited cell growth and induced apoptosis in lymphoma cell lines in a time and dose dependent manner. After 48 hours of incubation, the IC50 ranged between 50 and 100 nM, and it was equally effective in ABC and GCB-derived DLBCL cell lines. NVP-BEZ235 induced cell death was primarily due to induction of apoptosis, as evident by the annexin-V and PI dual staining method, and the induction of caspase 3 and PARP cleavage. NVP-BEZ235 effectively inhibited the activation of the PI3K pathway at several steps, including decreasing the phosphorylation level of p-Akt (Ser473), p-Akt (Thr308), p-mTOR, p-4-EBPI and pP70S6K. Because lymphoma cells frequently depend on multiple activated signaling pathways to promote their survival, including the JAK/STAT pathway, we investigated the potential synergy between PI3K and JAK/STAT pathway inhibitors. Lymphoma cells were variably sensitive to the JAK1/2 inhibitor INCB16562 in vitro. Submaximal concentrations of NVP-BEZ235 demonstrated a synergistic activity with INCB16562. Collectively, our data show that the PI3K/mTOR inhibitor NVP-BEZ235 is highly effective against a wide range of lymphoma cell lines, and warrants evaluating it alone and in combination with JAK/STAT inhibitors in phase I/II clinical trials in patients with relapsed lymphoma.

Disclosures:

No relevant conflicts of interest to declare.
Pediatric Myelodysplastic Syndrome In Japan and China According to 2008 WHO Classification
Authors: Yang Wenyu , Zhang Peihong , Hama Asahito , Ito Masafumi , Kojima Seiji , Zhu Xiaofan
Published: October 2019

Abstract 5030

Unlike adult myelodysplastic syndrome (MDS), most of children with MDS have hypocellular bone marrow. Thus, distinguishing MDS from aplastic anemia (AA) can be diagnostically challenging in these patients. To clarify the frequency of pediatric MDS and AA with hypocellular bone marrow in eastern Asia, Japanese and Chinese pediatric hematologists and pathologists conducted a review of 100 patients with cytopenia (50 from Japan, 50 from China). Films stained with both peripheral blood (PB) and bone marrow (BM) along with specimens of BM trephine biopsy were available for all of the patients examined. Patients included 45 males and 55 females, with a median age at diagnosis of 9 years (range: 1 to 19 years). A total of 76 patients had pancytopenia at the time of diagnosis, while 23 had bi-cytopenia and 1 had isolated thrombocytopenia. The medium internal from onset to diagnosis was 319(1–4165) days. Most children present with nonspecific symptoms as a result of cytopenia, including pallor(24%), fatigue(11%), petechiae (27%), bleeding (31%), fever (19%) and infections (10%). Hepatosplenomegaly is uncommon. Before diagnosed, 7% of patients had idiopathic thrombocytopenic purpura (ITP) and 8% of patients had other diseases such as asthma, pneumonia and etc. Cytogenetic analyses performed in 75 patients indicated the karyotype was normal in 45 patients, while 4 had trisomy 8, 1 had monosomy 19, and 2 had trisomy 2. Other abnormalities found included t(X;3)(p11.2;q13), add(6)(p21), and +6. The final diagnoses reached by consensus among the hematologists for the 100 patients included AA in 30 cases, refractory cytopenia of childhood (RCC) in 59 cases, and refractory cytopenia with multi lineage dysplasia (RCMD) in 11 cases. The distribution of the subtypes in the Japanese and Chinese cases were 14:16 for AA, 33:26 for RCC, and 3:8 for RCMD, respectively. In 9 of the patients, the hematologists and pathologists could not agree on the final diagnosis. In 5 of these cases, the hematologists diagnosed the patients as AA, while the pathologists diagnosed RCC. We analyzed the difference between pediatric hematologists and pathologists are the quality of bone marrow and biopsy samples' disparity. Overall, our results suggest that there is no difference in the distribution of the MDS subtypes between Japanese and Chinese children. In order to ensure there is a correct diagnosis of childhood MDS, cooperation between hematologists and pathologists is indispensable.

Disclosures:

No relevant conflicts of interest to declare.
Impact of Age on Physician's Decision to Pursue Active Treatment in Elderly Patients with Myelodysplastic Syndromes: Results from an Internet-Based Survey
Authors: Smith B. Douglas , Santini Valeria , Kurtin Sandra E. , Huber John , Campbell Kenneth , Waldman Anita
Published: October 2019

Abstract 5045

PURPOSE:

Myelodysplastic syndromes (MDS) are a group of hematological disorders predominantly affecting older patients. A patient's age influences the treating physician's decision to proceed with active treatment in MDS, with older patients being less likely than younger patients to receive active treatment, irrespective of their physical status.

DESIGN:

This study is an internet-based survey of hematologists and oncologists who treat patients with MDS consisting of a 47-item questionnaire designed to assess 1) factors physicians consider when determining treatment recommendations for MDS patients, 2) the impact of patient age on treatment decisions and 3) views on tools available to physicians to guide their treatment plans. Respondents were given hypothetical patient scenarios and asked to select their recommended treatment based on the available information.

RESULTS:

A total of 310 online interviews were conducted in North America and Europe. Although physicians cited biological age and disease performance status as more important, chronological age nevertheless emerged as a strong influencing factor on their treatment recommendations for MDS patients aged 65 years and older. For instance, physicians were significantly more likely to prescribe active therapy to a 70-year-old patient with secondary MDS and an intermediate International Prognostic Scoring System level than to an 80-year-old patient with otherwise identical characteristics (32% vs. 12%; p<0.05). Concerns regarding treatment toxicity influenced the recommendations of 66% of physicians, and 29% expressed dissatisfaction with the amount of data available to help guide treatment recommendations. A majority (62%) agreed that better guidelines would prevent under-treatment of older patients.

CONCLUSION:

Chronological age can influence physicians' treatment recommendations for MDS patients. The findings suggest that some older patients may be denied appropriate treatment options due to age and support the need for additional clinical evidence for older patients with MDS, such as validated comprehensive geriatric assessment tools to support treatment decisions regarding active treatment.

Disclosures:

Smith: Celgene: Honoraria. Santini:Celgene: Honoraria. Kurtin:Celgene: Honoraria. Huber:Celgene: Honoraria. Campbell:Celgene: Honoraria. Waldman:Celgene: Honoraria.
The Limited Screening Value of Insulin-Like Growth Factor-I in 610 Very Long Term Adult Survivors of Leukemia and Other Types of Childhood Cancer
Authors: Blijdorp Karin , van den Heuvel Marry M. , Pieters Rob , Boot Annemieke , van der Lely Aartjan , Neggers Sebastian J.C.M.M.
Published: October 2019

Abstract 506FN2

Introduction

Over the last decades childhood cancer survival rates have improved significantly. Currently, 70–80% of patients become long term cancer survivors. It has been estimated that 1 out of 640 young adults in the U.S. is a survivor of childhood cancer. Consequently, the incidence of late, treatment-related complications is increasing. Endocrine sequelae, such as the metabolic syndrome, osteopenia, sub fertility, thyroid dysfunction and growth hormone deficiency represent an important category of such late effects. Growth hormone deficiency (GHD) in childhood cancer survivors, mainly caused by cranial radiotherapy, is reflected by low levels of insulin like growth factor 1 (IGF-I). Whereas the value of IGF-I measurement for the diagnosis of GHD is controversial, low IGF-I levels are associated with high body mass index and high visceral fat percentage. However the clinical relevance of low IGF-I levels in long term childhood cancer survivors is not extensively studied. In this study we evaluated whether IGF-I is useful as a marker for altered body composition and growth hormone deficiency in this group.

Methods

We retrospectively analyzed data of 610 adult childhood cancer survivors, retrieved from the Rotterdam late effects clinic, which starts 5 years after cessation of therapy. Median age at diagnosis was 6 years (interquartile range (IQR) 3–11) and follow up time was 18 years (13–24). We assessed IGF-I Z-scores, anthropometrical measures, growth hormone stimulation tests in patients with clinical suspicion of GHD and measures of body composition (assessed by dual X-ray absorptiometry, Lunar Prodigy).

Results

Cranial irradiated acute leukaemia survivors (25 Gy (24–25)) and locally irradiated brain tumour survivors (42 Gy (35–54)) had significantly lower IGF-I Z-scores (p<0.001), lower height SDS (p<0.001), higher body mass index (p=0.01), higher waist-hip ratio (p=0.001), higher visceral fat percentage (p<0.001), higher total fat percentage (p<0.001) and lower lean body mass (p<0.001), compared to non cranial irradiated survivors. IGF-I did not show a strong correlation with BMI (r=-0.12, p=0.04), waist hip ratio (r=-0.15, p=0.01), total fat percentage (r=-0.14, p=0.02) and lean body mass (r=0.15, p=0.01). In the patients with low IGF-I levels who had growth hormone stimulation, IGF-I Z-scores did not significantly differ between the patients with and without severe GHD (p = 0.39).

Conclusion

This study shows that there is a limited value of IGF-I as marker for alterations in body composition, and confirms the fact that low IGF-I levels are not predictive for GHD, in a large cohort of childhood cancer survivors. Therefore the use of IGF-I should not be encouraged in adult childhood cancer survivors.

Disclosures:

No relevant conflicts of interest to declare.
Gynecomastia In Multiple Myeloma Patient Treated with Cytoxan
Authors: Jajeh Ahmad
Published: October 2019

Abstract 5131

Cytoxan is an active drug in the treatment of multiple myeloma. It is commonly reserved for patients who became refractory to common active drug therapy or in combination with other active agents such as bortezomib, pegylated liposmal doxorubicin, vincristine, melphalan and decadron in patients who are not eligible for autologous bone marrow transplant. This abstract is intended to present interesting side effect of cytoxan that seen only in multiple myeloma and not seen in other hematological malignancies or solid tumors. Gynecomastia is defined as hypertrophy of breast tissue in male which can be seen in various medical condition associated with endocrine disturbances. In this abstract will describe five cases of significant degree of gynecomastia that seen exclusively in african american males and not seen in white males. Median age of patients is 57 years, range 40–68. Three has failed tandem autologus bone marrow transplant and has been exposed to lenalidomide and bortezomib. The other two have failed many lines of therapy including melphalan and prednisone. Gynecomastia has not been seen in other white males who received cytoxan in the treatment of multiple myeloma. The degree of gynecomastia was very significant in all of the five african american males. Also this is a limited number of patients but it is interesting enough to draw attention on possible endocrine disturbances seen particularly in multiple myeloma and other plasma cell dyscrasia such as Poems syndrome. The fact that gynecomastia is exclusively seen in african american males who all denied abusing marijuana is of scientific interest that needed further study and attention.

Disclosures:

No relevant conflicts of interest to declare.
Alternative Dosing of Cyclophosphamide, Bortezomib and Corticosteroids (CyBorD) for Relapsed/Refractory Multiple Myeloma
Authors: Ho Viet Q. , Finley-Oliver Elizabeth , Shain Kenneth H. , Alsina Melissa , Ochoa Leonel , Nishihori Taiga , Djulbegovic Benjamin , Baz Rachid
Published: October 2019

Abstract 5142

Background:

The combination of an alkylating agent and bortezomib has been shown to be synergistic both preclinically and in the clinical arena. Although the combination of cyclophosphamide, bortezomib and corticosteroids (CyBorD) has been evaluated by several investigators, many have used different doses and schedule of cyclophosphamide and an optimal schedule has not been established. Herein, we report the safety and efficacy of this combination in patients with relapsed and refractory multiple myeloma.

Methods:

All patients with myeloma that received this schedule of CyBorD at the Moffitt Cancer Center were included. CyBorD was comprised of cyclophosphamide 1000 mg/2 IV on Day 1, Bortezomib 1.3 mg/m2 IV on Days 1, 4, 8, and 11 and either low dose dexamethasone (equivalent of 120– 160mg/cycle) or prednisone (100 mg PO day 1–5). Demographic, laboratory and clinical data was collected and evaluated via descriptive statistics. Response and progression are defined as per the IMWG uniform response criteria.

Results:

A total of 20 patients were identified and included for analysis. Demographic data is shown in table 1. At baseline, 35% of patients were platelet transfusion dependent, and the median ß2m was 6.2mg/L. 8 patients (40%) received prophylactic GCSF and 9 patients (45%) received secondary GCSF after significant neutropenia was noted. Overall response rate (PR and better) was 50% (70% minimal response and better), including 1 complete response (CR) and 5 very good partial response (VGPR). Median progression free survival was 3.4 months (95% CI 0–7.5 months) and median overall survival was 11.9 months (95% CI 3.3–20.0). The incidence of grade 3/4 neutropenia and thrombocytopenia was 80% and 75%, respectively. Grade 2 and 3 peripheral neuropahty developed in 1 and 1 patient respectively. Nine patients required dose reductions of cyclophosphamide (only 2 of the 8 who received prophylactic GCSF therapy) and 3 patients required dose reductions of the bortezomib.

Conclusion:

In this cohort of patients with high burden of disease, intermediate dose cyclophosphamide administered once every 3 weeks in combination with standard dose and schedule of bortezomib results in a meaningful response rate (albeit short PFS) with mainly hematologic toxicities that are manageable with growth factor support.

Disclosures:

Off Label Use: Cyclophosphamide in combination with bortezomib for relapsed/refractory multiple myeloma. Finley-Oliver:Millenium: Speakers Bureau. Alsina:Millennium: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees; Allergan: Research Funding. Djulbegovic:Millenium: Research Funding. Baz:Millenium: Research Funding, Speakers Bureau; Celgene: Research Funding.
The Phenotypic Profile of Circulating Microparticles Is Different in Polycythemia Vera and Secondary Erythrocytosis
Authors: Charpentier Agnès , Devaux Stéphanie , Bruge-Debreu Judith , Cambier Nathalie , Rose Christian , Demory Jean-Loup
Published: October 2019

Abstract 5166

Rationale:

In Ph1 myeloproliferative neoplasms (MPN), the blood stream carries a variety of microparticles and exosomes released by cells from myeloid lineages and endothelium; we had already demonstrated an excess of platelet microparticles (PMP) in ET and PV patients (ASH Annual Meeting 2009, Abstr. 1906). A further step has been to characterize five categories of MP depending on their origin. Then we used this method to compare the sub-populations found in Polycythemia vera (PV) and secondary erythrocytosis (SE).

Patients and methods:

We analyzed plasma samples from 20 PV diagnosed in accordance to WHO classification and 20 SE patients. SE was defined by elevated hematocrit, normal or increased serum EPO, Jak2V617F negative and a presumable cause, mainly respiratory. The 2 groups were comparable for age and sex-ratio. Mean Hb level was 18.0g/dl in the PV group and 18.1g/dl in the SE group. The concentration of MP was measured by flow cytometry using a FC500 cytometer (BeckmanCoulter™). All the MP subtypes express Annexin V thus defining the total MP. Additional co-expression of either CD41, CD144, CD14, CD11b or GPA identify MP populations originating respectively from platelets (PMP), endothelial cells (EMP), monocytes (MoMP), granulocytes (GrMP) and red cells (EryMP). Pre-analytical and testing procedures complied with the recommendations of the ISTH Standardization Sub-committee.

Results:

MP from endothelial cells (EMP) and leukocytes (GrMP and MoMP) are minor populations whose proportions are similar in both groups of patients (7%, 4% and 2% respectively). Conversely, EryMP and PMP are significantly different (p<0.001 Kursall-Wallis test) in PV and SE as shown in the following table.

So, in PV most of the MP are PMP while EryMP widely predominate in SE; in an attempt to explain such unexpected results we postulate that the cells generating MP are more or less mature depending on the physiopathology: clonal hematopoietic progenitors in MPN and well-differentiated cells in reactive conditions. However, even in SE, the EryMP concentration does not correlate with RBC count, haemoglobin or hematocrit.

Conclusion:

This study demonstrates a different phenotypic profile of circulating MP in PV and SE, possibly helpful to precise the origin of an elevated hematocrit, especially when molecular markers are negative. The significance of the MP subtype pattern observed in PV and SE remains unclear, calling for further studies.

Disclosures:

No relevant conflicts of interest to declare.
Tubastatin A, a Selective HDAC6 Inhibitor, Enhances Antigen-Presenting Cell (APC) Function and Restores the Responsiveness of Anergic CD4+ T Cells
Authors: Wang Hongwei , Wang Zi , Cheng Fengdong , Woan Karrune V. , Rock-Klotz Jennifer , Sahakian Eva , Powers John , Javier Pinilla , Bergman Joel , Villagra Alejandro , Kozikowski Alan , Sotomayor Eduardo M.
Published: October 2019

Abstract 520

Histones play a critical role in transcriptional regulation, cell cycle progression and developmental events. Histone acetylation/deacetylation alters chromatin structure and affects transcription factor access to DNA. Histone deacetylase inhibitors (HDI) induce growth arrest, cellular differentiation, and apoptosis and are being pursued as anticancer drugs. Interestingly, in addition to their antitumor properties, HDI have also been shown to modulate inflammatory responses.

Recently, we have found that HDAC6 is required for the production of the immunosuppressive cytokine IL-10 by APCs. Given the role of this cytokine in T-cell tolerance induction we asked whether inhibition of HDAC6 with Tubastatin A, a potent and selective HDAC6 inhibitor would influence the inflammatory status of APCs and their ability to determine activation versus tolerance of antigen-specific CD4+ T cells in vitro. First, treatment of peritoneal elicited macrophages (PEM) with increasing concentration of Tubastatin A resulted in enhanced tubulin acetylation which was accompanied by enhanced expression of co-stimulatory molecules in treated cells. In addition, Tubastatin-A treated PEM were unable to produce IL-10 and TNF-a in response to LPS stimulation. In sharp contrast, Tubastatin-A treated PEM produce higher level of the pro-inflammatory cytokines IL-12 and IL-6. Next, we evaluated the ability of Tubastatin A-treated APCs to present cognate antigen to naïve and tolerant CD4+ T-cells specific for a MHC class II restricted epitope of influenza hemagglutinin (HA). We found that treatment of PEM with Tubastatin A significantly enhanced their antigen-presenting capabilities leading to effective priming of naïve CD4+ T-cells confirmed by their increased production of IL-2 and IFN-g in response to cognate antigen. More importantly, Tubastatin-A treated APCs were able to restore the responsiveness of tolerant CD4+ T cells isolated from lymphoma bearing mice.

Taken together, selective HDAC6 inhibition with Tubastatin A provides a novel therapeutic approach to induce inflammatory APCs and overcome the significant barrier that T-cell tolerance has imposed to effective lymphoma immunotherapy.

Disclosures:

No relevant conflicts of interest to declare.
Establishment of An Angioimmunoblastic T-Cell Lymphoma (AITL) Model in the NOG Mouse
Authors: Ito Asahi , Ishida Takashi , Sato Fumihiko , Mori Fumiko , Masaki Ayako , Ri Masaki , Kusumoto Shigeru , Komatsu Hirokazu , Inagaki Hiroshi , Iida Shinsuke , Ueda Ryuzo
Published: October 2019

Abstract 5207

The aim of the present study was to establish a murine AITL model. We inoculated cells from affected LN of an AITL patient intraperitoneally into NOG mice. Hepatosplenomegaly developed in these animals about 2 months later, and normal splenic architecture was replaced by multi-focal deposit of lymphocytes and numerous blood vessels. Some of the former consisted of AITL cells characterized by small to medium size, and clear to pale cytoplasm. The remaining lymphocytes were non-neoplastic reactive cells including CD8-positive cells, B cells, and plasma cells. Double immunostaining revealed that the neoplastic cells were positive for both UCHL-1 (CD45RO) and BCL-6. In addition, significant levels of human IgG/A/M were detected in these animals. The AITL cells engrafted in the NOG mice indeed functioned as follicular helper T (Tfh) cells and induced antibody production by B-cells, consistent with recent evidence that AITL is a neoplasm originating from Tfh cells. These clinical and histological features in the mice are almost identical to those seen in AITL patients. Cells from spleens of affected animals could be serially transplanted, with enrichment of the AITL cells together with reduction of the reactive cells at each passage. This phenomenon might reflect the progressive nature of AITL. TCRB analysis demonstrated that the AITL clone in the mice was identical to that from the donating patient. This is the first mouse model of AITL, and could be a powerful tool for investigating, and developing novel treatment modalities for this type of lymphoma.

Disclosures:

No relevant conflicts of interest to declare.
Stage I or II Marginal Zone B-Cell Lymphoma; Multicenter Retrospective Analysis of 217 Cases
Authors: Oh Sung Yong , Kim Won Seog , Suh Cheolwon , Hong Junshik , Lee Soon Il , Song Moo-Kon , Kim Jung Han
Published: October 2019

Abstract 5219

Background:

Local stage- Ann Arbor stage I/II- Marginal zone B-cell lymphoma (MZL) was occupied 50∼70% of all MZL patients. Even though almost of them were well controlled with local treatment based therapy, But there is no data about risk factors and role of additional chemotherapy in stage II after local treatment. In this study, we conducted multicenter retrospective analyses of local stage MZL in order to assess their clinical features, optimal treatments, and prognoses.

Methods:

Between May 1992 and Jan. 2011, a total of 217 patients with biopsy-confirmed Ann Arbor stage I/II -MZL from 7 different institutions in Korea were included for analysis in this study.

Results:

The median age of our subjects was 48 (range: 14–82) years. This study involved 102 males (47%) and 115 females (53%). The most common primary sites of involvement were the orbit and ocular adnexa (43%) followed by small or large bowel (14%), and nodal MZL (10%). 190 patients (88%) and 27 patients (12%) were stage I and II, respectively. Of 217 patients with stage I or II, 213 patients were available for treatment outcome analysis. 167 (77%) were treated with local modality such as radiation therapy or surgery alone, 24 patients (11%) received additional systemic chemotherapy after local treatment. 26 patients (12%) were treated with chemotherapy alone. The overall response rate was 98% (95% CI, 96–100%) with 188 CRs and 20 PRs. Median progression free survivals (PFS) was 10.5 years (95% CI, 7.5–13.4 years). Overall survival (OS) did not reach the median value. 5 year and 10 year OS rates were 94% and 88%, respectively. During the follow-up 38 relapse events were observed. Poor prognostic factors influence to PFS were stage II (P=0.007) and chemotherapy alone treatment (P=0.002).

Conclusion:

Local stage- Ann Arbor stage I/II- MZL had been observed a good clinical course and well controlled with local treatment modality without additional chemotherapy. But, stage II MZL patients who need chemotherapy had relatively poor survival.

Disclosures:

No relevant conflicts of interest to declare.
The Nup98-HoxA9 Leukemogenic Fusion Protein Blocks Xpo1 and Tap-Mediated Nuclear Export
Authors: Dogan Yildirim , Sahay Sutapa , Dorn David C , Franceschino Anna , Xu Michael , Moore Malcolm A.S.
Published: October 2019

Abstract 5228

In leukemia Nup98 fusion genes are found as a product of the reciprocal translocation between Nucleoporin 98 (nup98) and homeobox-cluster genes. Nup98-HoxA9 (NHA9), the product of the t(7;11) translocation, is detected in acute myeloid leukemias (AMLs) and as a secondary abnormality in blast crisis of chronic myeloid leukemia (CML). The role of NHA9 in leukemogenesis is complex and incompletely understood. Here, we show an abrogation of nucleocytoplasmic shuttling of the nuclear export receptor Xpo1 and Tap in NHA9-expressing cells by using retroviral nuclear trafficking as a model. Lentiviral Rev, the prototype for nuclear export signal (NES)-containing proteins is shuttled through the nucleopore by Xpo1. NHA9 sequestered Xpo1 from the nuclear rim into nuclear aggregates resulting in deficient Xpo1-dependent nuclear exit of Rev and its mRNA substrates. Tap is involved in mRNA nucleocytoplasmic shuttling and is also responsible for the nuclear export of D-type retrovirus CTE-mRNAs. By using Tap/CTE-mRNA nuclear export as a model we also found that Tap colocalized in NHA9 nuclear aggregates leading to impaired Tap-mediated nuclear exit of CTE-mRNA substrates. Leukemogenicity of Nup98 fusion proteins may be accounted for in part by defects in Tap and Xpo1-mediated export of their substrates.

Disclosures:

No relevant conflicts of interest to declare.
Molecular and Pathological Analysis of DOCK Family Proteins in Hematopoietic Malignancy
Authors: Nishihara Hiroshi , Wang Lei , Nishio Mitsufumi , Endo Tomoyuki , Fujimoto Katsuya , Obara Masato , Tanaka Shinya
Published: October 2019

Abstract 5230

Crk, originally identified as an oncogene product v-Crk encoding in a chicken retrovirus CT10 in 1988, is a signaling adaptor protein mostly composed of SH2 and SH3 domains, and has been shown to play a pivotal role in cell proliferation, differentiation, and migration in various human cancers. Crk SH3 domain binds to DOCK180 which has been reported to activate Rac regulating cytoskeletal reorganization leading to the enhancement of cell motility and engulfment. In 1999, we identified DOCK2, a hematopoietic cell-specific homolog of the DOCK family protein, and revealed that DOCK2 regulated cell motility and cytokine production through the activation of Rac in human hematopoietic cells (1–3). Recently, we demonstrated the prominent expression of DOCK2 in human B cell lymphoma, and DOCK2 regulated cell proliferation through Rac and ERK activation in B cell lymphoma cell lines (4). Immunostaining for DOCK2 in 20 cases of human B cell lymphoma tissue specimens including diffuse large B cell lymphoma and follicular lymphoma revealed the prominent expression of DOCK2 in all of the lymphoma cells. DOCK2-knockdown (KD) of the B cell lymphoma cell lines, Ramos and Raji, using the lentiviral shRNA system presented decreased cell proliferation compared to the control cells. Furthermore, the tumor formation of DOCK2-KD Ramos cell in nude mice was significantly abrogated. Western blotting analysis and pull-down assay using GST-PAK-RBD chimeric protein suggested the presence of DOCK2-Rac-ERK pathway regulating the cell proliferation of these lymphoma cells. In addition, we confirmed the elevated expression levels of DOCK2 in various types of hematopoietic tumor cell lines as well as in the primary cells from the patients with leukemia and lymphoma. Interestingly, we also found the expression of DOCK180, not DOCK2, in Hodgkin/Reed-Sternberg cells of Hodgkin lymphoma tissue specimens as well as in three cell lines derived from Hodgkin lymphoma. We are currently analyzing the tumorigenic roles of DOCK2 and DOCK180 using the suitable cell lines derived from various hematopoietic neoplasms.

Here we summarize the results of our molecular and pathological analysis for the DOCK family proteins in various human hematopoietic malignancies such as lymphomas and leukemia. Our results suggest the possible target molecules of DOCK family proteins including DOCK2 and DOCK180, and also Rac for lymphoma and leukemia therapy.

Disclosures:

No relevant conflicts of interest to declare.
Loss of Heterozygosity (LOH) of PTPRJ in Non-Hodgkin`s Lymphoma (NHL)
Authors: Aya-Bonilla Carlos A , Green Michael R , Camilleri Emily , Marlton Paula , Lea Rod , Gandhi Maher , Griffiths Lyn R
Published: October 2019

Abstract 5231

The identification of Tumor Suppressor Genes (TSGs) in tumor tissue lacking of its normal germ-line counterpart has been limited due to the absence of proper statistical approaches that allow inferring the specific germ-line heterozygous status of a patient. Nonetheless, the implementation of Hidden Markov model (HMM) or cohort heterozygosity comparisons (CHC) approaches in the analysis of SNP array data from un-paired tumor DNA samples have permitted the detection of LOH regions and possible TSGs that might be implicated in the origin and development of cancer. Non-Hodgkin's Lymphoma (NHL) is the most common type of lymphoma. Nevertheless, its aetiology and pathogenesis remain unclear because of heterogeneity in biology, genetics and chemoresponse.

Here, we employed high-density SNP array analysis of tumors from two of the most frequent subtypes of NHL, Follicular Lymphoma (FL) and Diffuse Large B-cell Lymphoma (DLBCL). This revealed a high frequency of LOH over the chromosomal region of 11p11.2, containing the gene encoding the receptor-type protein tyrosine phosphatase, PTPRJ. LOH of this gene has been previously described in colorectal, lung, thyroid and breast cancer and the gene product regulates key survival pathways in B-cells, such as MAPK, PLCγ1, Scr and BCR signalling.

We validated LOH of the PTPRJ region by microsatellite analysis of an extended cohort of NHL tumors and found significantly increased rates of homozygosity compared to a patient-matched control population. Furthermore, sequencing of NHL cases revealed a strong association between LOH and the presence of a non-synonymous SNP in the PTPRJ coding region. Together, these results show the utility of LOH screening in identifying novel tumor suppressor genes, and implicate LOH of the PTPRJ gene as a potential lymphomagenic mechanism.

Disclosures:

Aya-Bonilla: Cancer Council of Queensland: Research Funding.
The AG Genotype of the Wilms Tumor-1 rs16754 SNP Is Associated with Poor Outcome in Pediatric AML Patients Treated with Stem Cell Transplantation but Not in Adults
Authors: Abalkhail Hala , El-Solh Hassan , Alseraihy Amal , Ayas Mouhab , Al-Ahmari Ali , Belgaumi Asim F , Al-Jefri Abdullah , Altombakty Ola , Toulimat Mohammed , Almhareb Fahed , Chaudhri Naeem A. , Walter Claudia Ulrike , Aljurf Mahmoud , Albitar Maher
Published: October 2019

Abstract 5237

The synonymous single nucleotide polymorphism rs16754 in the Wilms Tumor-1 gene (WT1) has been reported to correlate with outcome in adult patients with acute myeloid leukemia (AML) when treated with intensive chemotherapy. Specifically the GG genotype is significantly associated with favorable outcome, but also the AG genotype has been reported to be associated with better outcome as well. The clinical relevance of the rs16754 SNP in pediatric AML patients is far from clarified. In addition, it is not known whether allogeneic hematopoietic stem cell transplantation (HSCT) can modify the role of this SNP and its association with outcome.

Methods:

Genotyping of the SNP by direct sequencing of the exon 7 was performed on bone marrow samples from 86 AML patients (38 pediatrics and 48 adults). All patients were treated with HSCT. Most patients (39 adults and 28 pediatrics) were transplanted in first remission (CR1). The median age was 25 years (range: 14–54) for adults and 8 years (range: 8 months–14 years) for the pediatric group.

Results:

In pediatric AML, we detected the AA genotype in 22 patients (53%), the GG genotype in 2 patients (5%) and the remaining (42%) had AG genotype. In the adult patients, the AA genotypye was present in 26 patients (54%), GG genotype in 3 (6%) patients, and the AG genotype in the remaining patients (39%). A similar distribution was observed in the normal population (58%, 12%, 30%, for AA, GG, and AG, respectively). In pediatric patients, the AG genotype significantly correlated with shorter overall survival (OS) (P=0.04) and event free survival (EFS) (P=0.04) when compared with the patients with AA genotype. In contrast, in adult AML, groups with AG and AA geneotypes completely overlapped for OS and EFS. When only patients treated with HSCT in CR1 were considered, the pediatric patients showed the same trend (P=0.07), but there was no correlation with survival in the adult group. This analysis included patients with intermediate and adverse risk cytogenetics. AG genotype was not a predictor of outcome in multivariate model incorporating cytogenetics and the WT1 genotype. The patients with the GG genotype were too few for analysis.

Conclusion:

This data supports the concept that the biology of AML in pediatric patient is different from that in adults. The role of this synonymous SNP in AML needs further exploration, especially investigating the potential that this SNP may have some effects on the host and not only the disease.

Disclosures:

No relevant conflicts of interest to declare.
Evaluation of ADAMTS13 Activity and Inflammatory Markers in Deep Venous Thrombosis Patients
Authors: de Moraes Mazetto Bruna , Orsi Fernanda A. , Barnabé Aline , de Paula Erich V , Annichino-Bizzacchi Joyce M
Published: October 2019

Abstract 5246

BACKGROUND:

The role of inflammation on the pathophysiology of arterial thrombosis has been extensively studied. However, the relationship between inflammation and deep vein thrombosis (DVT) is not completely understood. Conflicting reports have been published about the levels of proteins involved in the inflammatory response in the context of DVT. Changes in ADAMTS13 levels have been reported in other inflammatory conditions such as sepsis, and this protease could be involved in the interplay between hemostasis and inflammation.

OBJECTIVE:

To evaluate ADAMTS13 activity in DVT patients and its association with inflammatory markers (IL-6, IL-8, CRP, TNF-α), D-dimer and von Willebrand Factor (VWF) levels.

METHODOLOGY:

Thirty-eight DVT patients, from 6 months to five years after the diagnosis of DVT, followed at the outpatient unit of thrombotic diseases from University of Campinas and 38 healthy volunteers selected as controls were included in the study. ADAMTS13 activity was determined by the residual binding of VWF to collagen. VWF, IL-6, IL-8 and TNF-alpha levels were determined by ELISA, and D-dimer levels was determined by a turbidimetric method.

RESULTS:

In this study population DVT was triggered by transient risk factors, particularly the use of oral contraceptives. No patient presented renal, hepatic or malignant disease. Median ADAMTS13 activity was not statistically different between patients (median: 98.2%; range: 70–146) and controls (median: 96.1%; range: 66–117; p=0.35). IL-6 and TNF-alfa levels were also higher in patients (median: 1.0pg/mL and 2.3pg/mL) compared to controls (median: 0.64pg/mL and 1.7pg/mL, p=0,01 and p=0,009). In addition, VWF and D-dimer levels were also significantly higher in patients (all P<0.01). No significant difference could be demonstrated between IL-8 and CRP levels from patients and controls. No significant correlation between ADAMTS13 levels and other inflammatory markers could be demonstrated. VWF levels correlated significantly with IL-8 (r=-0.4635, p< 0.0001).

CONCLUSIONS:

This study reinforces the role of inflammation in the pathogenesis of DVT. The correlation of VWF and IL-8 supports the notion that IL-8 stimulates the secretion of VWF. The role of ADAMTS13 in the context of DVT is yet to be determined.

Disclosures:

No relevant conflicts of interest to declare.
Analysis of the Genetic Expression of Inflammatory Mediators From Patients with Spontaneous Deep Venous Thrombosis
Authors: Barnabé Aline , Santiago-Bassora Fernanda Dutra , dos Santos Andrey , Carazzolle Marcelo Falsarella , Pereira Gonçalo AG , de Andrade Orsi Fernanda Loureiro , de Paula Erich V , Höehr Nelci Fenalti , Annichino-Bizzacchi Joyce M
Published: October 2019

Abstract 5251

Deep venous thrombosis (DVT) is a multifactorial disease and in about 30% of patients no risk factor can be identified. Association of inflammation and hemostasis is thought to play a significant role in the pathogenesis of DVT. We hypothesized that distinctive patterns of expression of inflammatory mediators in mononuclear cells from patients with previous DVT could be relevant to the pathogenesis of DVT. cDNA microarray technology (CodeLink Bioarrays) was chosen to study the gene expression profile of inflammatory mediators in DVT patients with clinical characteristics of a high penetrance of any putative hereditary risk, and their healthy siblings. These microarrays were hybridized with synthesized probes from pooled samples collected from patients and control siblings. Patients were divided into two clinical groups: (1) patients with one spontaneous DVT (n=2), and (2) patients with recurrent spontaneous DVT episodes (n=1). Preliminary analysis showed that 2% of approximately 55,000 transcripts contained in the array had significant (higher than 2-fold) differences in expression in spontaneous and recurrent spontaneous DVT relative to their asymptomatic siblings. In group 1, 738 genes were upregulated and 504 genes were downregulated. In group 2, 1229 genes were upregulated and 87 genes were downregulated. Genes with different expression compared to controls were associated to immune and inflammatory response, motility, cell adhesion, cell-cell signaling, defense response, signal transduction and apoptosis. Genes that presented the highest differences compared to controls were interleukin 8 (43-fold increase), chemokine (C-X-C motif) ligand 2 (11-fold increase), major histocompatibility complex, class II, DR beta 5 (8-fold increase), major histocompatibility complex, class II, DQ alpha 1 (29-fold decrease), caspase recruitment domain family, member 15 (7-fold increase), carboxypeptidase A5 (11-fold decrease), cathepsin G (8-fold decrease), and azurocidin 1 (3-fold decrease). A comprehensive list of these genes was generated. Evaluation of the significance of these distinctive gene expression profiles between patients with spontaneous and recurrent DVT compared to their asymptomatic siblings could reveal novel insights into the pathophysiology of the first DVT episode and of DVT recurrence, as well as new biomarkers or therapeutic targets.

Disclosures:

No relevant conflicts of interest to declare.