bluuebunny/crossref_metadata_2025 huggingface dataset:
Constitutive Lymphoid Expression of the Nuclear Form of RNase H1 Is Associated with Development of Immature B Cell Lymphomas.
Authors: Sakhuja Kiran B , Gugliotti Lina A , Suzuki Yutaka , Kovalchuk Alexander L , Wang Hongsheng , Shin Dong-Mi , Cerritelli Susana M , Morse Herbert C , Crouch Robert J
Published: October 2019

Abstract 1613

Poster Board I-639

Ribonuclease H1, which cleaves the RNA of RNA/DNA hybrids, plays a vital role in the replication of mitochondrial DNA during mouse embryogenesis, but its contributions to later mammalian development and adult biology are not known. In higher eukaryotes two isoforms of RNase H1, a nuclear form and mitochondrial form, are produced from a single transcript. To investigate the effects of constitutive high level RNase H1 expression, we generated transgenic (TG) mice, strain M27F7, with lymphocyte-specific over-expression of nuclear RNase H1. Transcripts in B cells from TG mice were ∼100-fold higher than in wild type B cells. Although there was no obvious phenotype in young mice, some of the older mice, especially females, developed lymphadenopathy, splenomegaly and mediastinal masses and were diagnosed histologically as having lymphoblastic lymphomas. Cell lines established from the tumors were examined by FACS, gene expression profiling and SKY as well as by FISH for Igh and Myc. By FACS criteria, the tumors were B cell lineage and were arrested at the pre-B to immature stage of differentiation. Expression profiles showed the lines to have increased expression of anti-apoptotic genes and genes promoting cell cycle progression. Chromosomal studies revealed no signs of genomic instability or translocations, although there were trisomies of chromosomes 3 and 15. Myc was structurally intact in all lines. These results suggest that RNase H1 plays a previously unappreciated role in early B cell development and that constitutive overexpression contributes to lymphomagenesis.

This work was supported by the Intramural Research Program of the National Institutes of Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development and National Institute of Allergy and Infectious Diseases.

Disclosures

No relevant conflicts of interest to declare.
Acute Myelogenous Leukemia Blasts Contribute to a Bone Marrow-Stroma in In Vivo NOD/SCID Mouse System..
Authors: Tashiro Haruko , Shirasaki Ryosuke , Oka Yoko , Sugao Toshihiko , Akiyama Nobu , Kawasugi Kazuo , Shirafuji Naoki
Published: October 2019

Abstract 1621

Poster Board I-647

Aims: Cancer stem cell theory has been developed, and whose precise characteristics have been reported. However, there have been no reports on the differentiation of cancer stem cells into the environmental stromal cells. We reported previously that non-adherent acute myelogenous leukemia (AML) cells were differentiated into myofibroblasts to create a microenvironment for proliferation of AML blasts in vitro. In this report we demonstrate that with severe combined immunodeficiency (SCID) mouse system AML blast cells also convert to myofibroblasts to form stroma in vivo.

Materials and Methods

Bone marrow cells were collected from informed AML (M2) patients who had chromosomal translocation of RUNX1 and ETO, from which mononuclear cells were separated with density-gradient sedimentation method. After discarded an adherent cell-fraction, the non-adherent mononuclear cells were injected to the 3.0 Gray-irradiated non-obese diabetes (NOD)/SCID mouse intravenously. For the inactivation of NK cells, anti-Asialo GM1 antibody was injected intra-peritoneally prior to the transplantation, and on each 11th day thereafter. Blood was collected to monitor Runx1 and ETO fusion transcript, and mice were sacrificed after chimeric mRNA was observed. Bone marrow cells were obtained, and sorted with anti-human CD133 antibody and -CD106 to select AML-derived human stromal myofibroblasts referred to the in vitro data. The isolated positive fraction was further cultured, and the biological and the molecular characteristics were analyzed.

Results and Discussion

When non-adherent AML (M2) blast cells were transplanted to NOD/SCID mice, cells were engrafted after 10 weeks. In murine bone marrow cells human stromal cells were identified, in which RUNX1 and ETO gene was fused with FISH analysis. When the parental AML blast cells were cultured on the expanded AML-derived myofibroblasts, AML cells grew extensively. These results indicate that AML cells can create their own microenvironment for proliferation in vivo.

Disclosures

No relevant conflicts of interest to declare.
The Varicella Zoster Virus Immunoglobulin G and the Varicella Zoster Virus Infection in Children with Cancer Under Chemotherapy.
Authors: Shim Ye Jee , Lee Kun Soo , Choi Yong-Mook
Published: October 2019

Abstract 1648

Poster Board I-674

Purpose

The varicella zoster virus (VZV) is known to be fatal especially for immunocompromised patients receiving chemotherapy for childhood cancer. So we investigated the change of varicella zoster virus immunoglobulin G (VZV IgG) before and during chemotherapy in patients who suffered from chicken pox (CP) or herpes zoster (HZ) to find the relationship between them.

Methods

We reviewed the transition of VZV IgG through medical records in children with cancer who experienced CP or HS during chemotherapy from 1998 to 2008 in Kyungpook National University Hospital, Daegu, South Korea.

Results

We checked VZV IgG not only when they were diagnosed as cancer but also when they were receiving chemotherapy for 66 patients (VZV IgG (+) : VZV IgG (-) = 40 : 26). Among initial VZV IgG positive group, 28 patients showed sustained positive IgG during chemotherapy, and 6 patients underwent the disease (CP : HZ = 1 : 5). On the other hand, 12 patients showed negative IgG while chemotherapy and 8 patients experienced the disease (CP : HZ = 4 : 4). The seronegative group after chemotherapy showed higher morbidity of CP or HZ compare to seropositive group (p=0.011). In the case of VZV IgG negative group at diagnosis, 8 patients presented seroconsersion to positive IgG and they had no disease of VZV. Among them, 18 patients who showed still negative IgG developed 4 (CP : HZ = 2 : 2). All together, the seronegative group revealed the tendency of higher morbidity (p=0.034). We treated them using intravenous acyclovir, and no mortality was observed.

Conclusion

Because of higher frequency of CP or HZ in VZV IgG negative group during chemotherapy, it is important to decide when and for whom to carry out VZV vaccination during chemotherapy through regular VZV IgG checking.

Disclosures

No relevant conflicts of interest to declare.
High-Dose Methotrexate, Rituximab, and Temozolomide (MRT) for Patients with Primary CNS Lymphoma (PCNSL).
Authors: Gerstner Elizabeth R. , Hochberg Fred H , Plotkin Scott R. , Eichler April F. , Batchelor Tracy T.
Published: October 2019

Abstract 1672

Poster Board I-698

Introduction

PCNSL is a rare form of non-Hodgkin lymphoma. High-dose methotrexate (HD-MTX) is the backbone of therapy but uncertainty remains about what additional chemotherapies should be added to HD-MTX to improve response rates.

Methods

After receiving IRB consent, we retrospectively evaluated patients with PCNSL treated at our hospital with combination M/R/T at initial diagnosis or at relapse. Patients were treated in 28-day induction cycles as follows: HD-MTX (8g/m2- dose adjusted based on creatinine clearance) on days 1 and 15; rituximab (375 mg/m2) on days 3, 10, 17, and 24; and temozolomide (150-200 mg/m2) on days 7-11. HD-MTX was given every 2 weeks until complete response (CR) and for 2 additional treatments followed by monthly maintenance treatments for 11 months. Rituximab was given weekly for a total of 8 weeks. Temozolomide was continued for 6 months after CR. Brain MRI was done after every other methotrexate treatment to assess response.

Results

Sixteen patients were treated between February 2006 and August 2009. Ten patients received MRT as first-line therapy at the time of initial diagnosis and 6 received MRT as salvage therapy at first or second recurrence. The median age of newly diagnosed patients was 58 (range 47-76) and of relapsed patients was 60 (range 46-76). CSF cytology was atypical in 5/15 patients who underwent lumbar puncture (4 first-line, 1 relapse). After first-line therapy, there were 9 CRs (median cycles to CR = 3) and 1 PR (patient still receiving induction treatment). After salvage therapy, there were 4 CRs (median cycles to CR = 4) and 2 PRs (both patients died while receiving treatment- 1 from an unrelated myocardial infarction). With a median follow-up of 10.3 months in the first-line group and 7.2 months in the relapse group, only 3 patients have progressed; one of whom had clinical progression after a PR and a second who relapsed in the skin. Treatment was well tolerated with reversible grade 4 transammonitis in 1 patient and grade 3 hematological toxicities in 8 patients. One patient experienced grade 4 thrombocytopenia related to temozolomide requiring dose reduction for subsequent cycles.

Conclusions

Combination MRT is well tolerated and resulted in a promising early response rate. The median number of cycles needed to achieve a CR for patients with newly diagnosed PCNSL was less than the 6 cycles we have previously reported for HD-MTX monotherapy suggesting that adding rituximab and temozolomide is beneficial to patients. This combination warrants further evaluation in larger scale, prospective studies.

Disclosures

Off Label Use: temozolomide and rituximab for PCNSL..
Inhibition of Class I PI3K Isoforms Restores the Sensitivity of Acute Myelogenous Leukemia Cells to Multi-Tyrosine Kinase Inhibitors in the Bone Marrow Microenvironment.
Authors: Shimada Akira , Niu Hongmei , Orwick Shelley , Inaba Hiroto , Rubnitz Jeffrey E , Campana Dario , Baker Sharyn D
Published: October 2019

Abstract 1734

Poster Board I-760

The bone marrow microenvironment can protect leukemic cells from cytotoxic agents, an effect that has been attributed to both soluble- and contact-mediated factors. Recently, we showed that 3 AML cell lines (MV4-11, THP1 and U937) became resistant to 3 promising TKIs (sorafenib, sunitinib and midostaurin) in a coculture model with mesenchymal cells (MSCs) (Shimada A. et al. Proceedings ASH 2008). In the current investigation, we show that 9 primary AML blast samples (including 7 with wild-type FLT3 and 2 with FLT3-interal tandem duplications) were less sensitive to 10 μM sorafenib in coculture with MSCs vs. suspension culture [mean percentage of total annexin V positive cells, 25% (range, 17-32%) vs. 36% (range, 31-47%)]. We explored potential mechanisms of resistance in AML cell lines. No alterations in drug intracellular accumulation, FLT3 receptor expression/phosphorylation, or acquired mutations in the FLT3, NRAS or KRAS genes were observed in AML cells in coculture with MSCs. However, we found that contact with AML cells enhanced the secretion of IL-6, IL-8, and VCAM-1 by MSCs. In parallel, multiple signaling pathways were activated in AML cells by MSCs including PI3K/AKT, STAT, ERK, and NFKB. Sorafenib-induced inhibition of phosphoproteins in the PI3K/AKT and ERK pathways was decreased in coculture vs. suspension culture. We evaluated PI103, a selective and potent inhibitor of class I PI3K isoforms, as a potential strategy to overcome TKI resistance in the microenvironment. In 3 AML cell lines, the antileukemic activity of PI103 in combination with each of the 3 TKIs was enhanced to a greater extent in coculture than in suspension culture, which was associated with more effective inhibition of both phospho-AKT and phospho-p70 S6 kinase. In primary AML blast samples, the combination of 1.0 μM PI103 + 10.0 μM sorafenib induced more apoptosis [mean percentage of total annexin V positive cells, 38% (range, 32-47%)] than either agent alone [sorafenib, 25% (16-32%); PI103, 24% (20-36%)]. In conclusion, inhibition of PI3K/AKT signaling represents a potential treatment strategy to restore TKI sensitivity in the microenvironment.

Disclosures

No relevant conflicts of interest to declare.
Polycomb Group Proteins in Myelodysplastic Syndromes and Its Transformation Into Acute Myeloid Leukemia.
Authors: Shen Hongjie , Chen Zi-Xing , Cen Jiannong , Wang Yuanyuan , He Jun
Published: October 2019

Abstract 1756

Poster Board I-782

The myelodysplastic syndrome (MDS) is a group of clonal blood disorders characterized by refractory cytopenia in peripheral blood, dysplastic cellular morphology in bone marrow, and a high risk of malignant transformation. Recently, expression of BMI1, a polycomb group protein, has been used as an important marker for predicting progression of MDS to acute myeloid leukemia(AML). However the function of BMI1 in MDS is remained unknown. Presently, we have analyzed the expression of SALL4, EZH2, P16 and BMI1 in bone marrow mononuclear cells from 48 newly diagnosed MDS patients and 40 treated MDS patients. Real-time polymerase chain reaction has shown that expression levels of these genes in MDS was frequently higher than that of non-MDS cytopenians (P<0.05). A high level of SALL4 expression in newly diagnosed MDS was always associated with high level of BMI1 expression. When MDS cases were divided into two groups based on their SALL4 mean level, The BMI1 expressed significantly higher in SALL4 higher group than that of SALL4 lower group (P<0.05). The BMI1 expression pattern showed an increasing tendency from MDS-RA moving to MDS-RAEB, without a statistically significance. On the contrary, the EZH2 expression level of MDS-RAEB is lower than that of MDS-RA (P<0.05). Although BMI1 expression level has been down-regulated when MDS patients are in morphological remission, the expression level of SALL4 and EZH2, P16 still remain abnormal (P<0.05). A significantly high level of BMI1 expression could usually be observed in MDS patients in progression, i.e. from RA to RAEB or from MDS to AML. To study whether BMI1 contributes to the MDS progression, we have transfected U937 and K562 cell line with BMI1 full-length cDNA mediated by MSCV retroviral vector and selected subclone cells. The viability of BMI1 transfected cell is much higher than the control cells cultured serum-free medium. Flow cytometry has shown that BMI1 transfected cell also inhibit apoptosis induced by As2O3. By real-time RT-PCR, it was found that BMI1 can repress the expression of EZH2, PTEN and P16 in BMI1 transfected U937 cell line, while only repress the expression of EZH2 in BMI1 transfected K562 cell line. These results suggest that the polycomb group family genes probably play a pivotal role in MDS progression, and BMI1 cooperated with EZH2 may act via an epigenetic mechanism in the pathogenesis of MDS.

Disclosures

No relevant conflicts of interest to declare.
Immunoglobulin D Multiple Myeloma: Clinical Presentation, Response to Therapy and Prognostic Factors in 75 Patients; An Analysis of the Korean Multiple Myeloma Working Party (KMMWP).
Authors: Kim Min Kyoung , Suh Cheolwon , Min Chang-Ki , Kim Seok Jin , Kim Kihyun , Moon Joon Ho , Yoon Sung-Soo , Lee Gyeong-Won , Kang Hye Jin , Kim Sung Hyun , Choi Chul Won , Eom Hyeon Seok , Kwak Jae-Yong , Kim Hyo Jung , Mun Yeung-Chul , Bang Soo-Mee , Lee Keehyun , Shin Ho Jin , Yang Deok Hwan , Lee Jae Hoon
Published: October 2019

Abstract 1792

Poster Board I-818

Purpose

The primary objective of this study was to analyze the clinical and laboratory characteristics, response to therapy, and survival in patients with IgD multiple myeloma. The secondary objectives were to evaluate the efficacy of treatment, including autologous stem cell transplantation, in these patients.

Patients and Methods

Records of 75 patients with IgD multiple myeloma from 18 institutions were reviewed using the Korean Myeloma Registry (KMR) database (www.myeloma.co.kr).

Results

Median patient age was 57 years (range, 34∼81 years) and 67% were male. The main presenting features were bone pain (77%) and fatigue (77%). Thirty patients (40%) had ECOG PS ≥ 2, and 71 (94%) had ≥1 lytic bone lesions. Renal function impairment (sCr level ≥2 mg/dL) and hypercalcemia (sCa level ≥12 mg/dL) were present in 53% and 27% of patients, respectively. Median serum M protein level was 1.4 g/dL, and 89% of patients had lambda light chains. Forty-nine patients (65%) were stage III by the ISS and 92% were stage III by DSS. Forty-nine patients were assessable for chromosomal analysis, 46 by conventional cytogenetics and 3 by FISH analysis. Of these, 23 patients (47%) had chromosomal abnormalities associated with poor prognosis. Thirty-nine patients (52%) were treated with VAD chemotherapy; the 36 patients assessable for response had an objective response rate of 59%. Eighteen patients (24%) received first-line combination chemotherapy including new drugs (bortezomib or thalidomide) and their objective response rate was 88%. At a median follow up of 28.6 months for the surviving patients, the median OS was 18.5 months and median PFS was 10.4 months. Multivariate analysis showed that age ' 65 (vs > 65), absence of extramedullary plasmacytoma, absence of del(13) or hypoploidy and serum beta-2 microglobulin level <8.0 mg/dL (vs ≥ 8.0mg/dL) were significant prognostic factors for OS. Thirty-four patients (45%) underwent SCT, 32 as part of their initial therapy. For patients who received SCT after complete or partial response to initial therapy, the median OS was 30 months.

Conclusion

IgD multiple myeloma is an aggressive disease that is usually detected at an advanced stage. Despite initial response, survival after relapse is dismal. Intensive treatment strategies to induce high quality responses before SCT may improve outcomes in younger patients.

Disclosures

Suh: Janssen Korea: Research Funding.
Molecular Characterization of Human Multiple Myeloma Cell Lines by Genome-Wide Profiling.
Authors: Elosua Carolina , Catalina Purificacion , Palma Cristina , Walker Brian A , Dickens Nicholas J , Gonzalez David , Davies Faith E , Morgan Gareth J , Leone Paola E.
Published: October 2019

Abstract 1793

Poster Board I-819

Multiple Myeloma (MM) is a malignancy depicted by clonal expansion of plasma cells in the bone marrow. There are two broad genetic subtypes of multiple myeloma as defined as hyperdiploid multiple myeloma (H-MM), characterized by trisomies of chromosomes 3, 5, 7, 9, 11, 15, 19, and 21, and nonhyperdiploid multiple myeloma (NH-MM) associated with primary translocations involving the immunoglobulin heavy chain (IgH). These two subtypes of multiple myeloma have two different molecular pathogenesis and characteristic changes of each have been already observed. In our study, we previously described the patterns of genetic lesions and molecular pathogenesis of 12 HMCLs with Affymetrix 500K Single Nucleotide Polymorphism-based mapping arrays, now we re-depict those patterns using the Illumina 1M array set and compare the two studies. These techniques allow the examination and identification of copy number changes, bi-allelic deletions and the identification of loss of heterozygosity (LOH) due to loss and uniparental disomy (UPD), as well as gene localization and identification. The 12 HMCLs utilized are characterized for their structural alterations and not by hyperdiploidy. In addition, so as to fulfill the selection criteria, a minimum of 3 cell lines must present the alterations cited below. Previously described gains were observed in 1q, 7q, 8, 11q, 18, 19, and 20q; but also found at 4q. The bi-allelic deletions were ascertained on 3p. Similarly, we identified the regions of mono-allelic deletions on 1, 2q, 6q, 8q, 9p, 11q, 12, 13q, 14q, 17p, and 20p. In addition, described regions of bi-allelic deletions were detected on 1p, 6q, 8p, 13q, 16q, and 22q, and furthermore located on 2q, 3, 4q, 9, 10q, 12p, and 20p. Finally, the UPD obtained were traced on 1q, 4q, 8q, 10q, and 22q. The use of the new platform has allowed us to re-identify, significantly increase in information above the original set and finely delimit the regions previously described. Taken together, the dysregulated genes from the myeloma genome indicate that the crucial pathways in myeloma include NF-kB, apoptosis, cell-cycle and critical intracellular signaling pathways including the JAK/STAT, Wnt signaling, RAS/RAF/MAPK and PI3k/AKT.

Disclosures

No relevant conflicts of interest to declare.
Molecular Profiling of Extramedullary and Medullary Plasmacytomas.
Authors: Mahindra Anuj , Amin Samir B , Motyckova Gabriela , Sohani Aliyah R. , Patel Kishan , Vallet Sonia , Chetri Gaurav , Ackerman Adam , Pozzi Samantha , Santo Loredana , Cirstea Diana , Toomey Christiana E. , Hochberg Ephraim P. , Cin Paola Dal , Hideshima Teru , Anderson Kenneth C. , Munshi Nikhil , Raje Noopur
Published: October 2019

Abstract 1806

Poster Board I-832

Plasmacytomas are rare clonal proliferations of plasma cells that though cytologically identical to plasma cell myeloma, present with osseous or extraosseous growth pattern. Understanding their molecular characteristics can provide crucial insights into their pathogenesis and risk of progression to multiple myeloma (MM). To investigate the differences between extramedullary (EMP) and medullary plasmacytomas (MP) and MM without plasmacytomas, we sought to molecularly profile these tumors by tissue microarrays, gene expression, microRNA, and FISH.

We identified 85 patients from our data base with a pathological diagnosis of plasmacytoma. Of the 85 patients, 13 patients presented with EMP, and 72 had MP. Among the patients with EMP (n=13), 2 patients presented with multiple lesions. Three of 13 (23%) patients progressed to develop MM at a median of 12 months. 72 patients presented with MP, of which 21 had solitary lesions and 27 (37%) progressed to MM at a median of 20.5months. There was a male preponderance (67% vs 33%) and the median age at diagnosis was 60.5 years (range 27.7-87.6). The mean overall survival for patients with EMP was 121 months (95% confidence interval[CI] 97-144 months) and for patients with MP was 102 months (95% CI 93-128 months) { p=0.025}

MicroRNA (miRNAs) profiling was performed on MP (n=19) and MM samples (n=66). Data was normalized using U6 endogenous control. Three hundred and one miRNAs out of a total 665 were significantly differentially expressed between MP vs MM samples. Gene expression profiling performed on MP will be correlated with the miRNA data to identify genes and transcripts of interest which will be functionally validated. Tissue microarrays were performed on 52 patients (8: EMP, 44: MP,) in whom paraffin-embedded tissue was available. Of samples analyzed, CD56 positivity was observed in 55% MP and 71% EMP samples (p=0.67). Additional staining for cyclin D1, Bcl 2 and FISH analysis will be reported.

Differential expression patterns of factors involved in proliferation, survival, adhesion, and stroma-tumor cell interactions may help explain plasmacytoma biology and identify factors responsible for progression to MM. These insights may help identify new therapeutic approaches and targets in the treatment of these plasma cell disorders.

Disclosures

Hochberg: Enzon: Consultancy, Speakers Bureau; Biogen-Idec: Speakers Bureau; Genentech: Speakers Bureau; Amgen: Speakers Bureau. Anderson:Millennium: Research Funding. Raje:Celgene, Norvartis, Astrazeneca: Research Funding.
The Effect of the CXCR4 Inhibitors, AMD3100 and AMD3465, in Multiple Myeloma Cell Homing and Its Interaction with the Bone Marrow Microenvironment.
Authors: Runnels Judith M. , Azab Abdel Kareem , Pitsillides Costas , Farag Jonathan , Wu Juwell , Roccaro Aldo M. , Maiso Patricia , Lin Charles P. , Ghobrial Irene M.
Published: October 2019

Abstract 1826

Poster Board I-852

Patients with multiple myeloma (MM) typically present with the disease spread diffusely throughout the bone marrow (BM). This fact points to the role MM cell trafficking plays in disease progression. Like normal plasma cells, MM cell trafficking is directed by the cytokine, SDF-1 and its receptor, CXCR4. Using the small bicyclam molecule, AMD3100, to block the SDF-1/CXCR4 interaction, and in vivo flow cytometry to measure circulation time of MM cells in a xenograft model, we have previously shown that homing of injected AMD3100-treated MM cells to the bone marrow is perturbed. Soon after injection fewer AMD3100-treated than untreated MM cells are detected in mouse skull BM using in vivo fluorescence confocal microscopy. Furthermore, the combined treatment of established tumors with AMD3100 and bortezomib enhanced survival. Here we present data that the 2nd generation SDF-1/CXCR4 small monomacrocyclic inhibitor, AMD3465 behaves like its predecessor in blocking antibody binding to CXCR4 on MM cells, migration of MM cells toward SDF-1, and in vivo homing, but at concentrations 10-50 fold lower than AMD3100. Incubation with either 1uM AMD3465 or 50uM AMD3100 reduced antibody binding to CXCR4 on MM cells to isotype control levels. Likewise, treatment with AMD3465 reduced MM1S migration to 15% of that of untreated cells in a transwell migration assay. These in vitro effects translated in vivo into longer circulation times for AMD3465-treated MM cells than control cells. In vivo flow cytometry revealed that 40-50% of AMD3465-pre-treated cells remained in the circulation one hr after injection whereas untreated cells depleted to 20% of their original circulating cell count by that time. Future experiments will further describe the effect of AMD 3465 on MM cells in the BM microenvironment to answer whether AMD3465 can be used to mobilize MM cells from the BM environment or enhance survival when used in conjunction with therapeutic drugs.

Disclosures

Ghobrial: Millennium: Honoraria, Research Funding, Speakers Bureau; Celgene: Consultancy, Honoraria, Speakers Bureau; Novartis: Honoraria, Speakers Bureau.
Impact of ABO Imcompatibility On Acute GvHD and Thrombotic Microangiopathy After Reduced-Intensity Cord Blood Transplantation.
Authors: Tsuji Masanori , Wake Atsushi , Uchida Naoyuki , Ishiwata Kazuya , Nakano Nobuaki , Takagi Shinsuke , Yamamoto Hisashi , Mori Yuki , Matsuno Naofumi , Masuoka Kazuhiro , Makino Shigeyoshi , Yoneyama Akiko , Taniguchi Shuichi
Published: October 2019

Abstract 2298

Poster Board II-275

Introduction:

Althouth ABO blood type is one of two antigen system for transplantation, the effect of ABO incompatibility on transplantation outcome still remains controversy. Furthermore, there is little data about ABO incompatibility on the outcome of unrelated cord blood transplantation following reduced-intensity conditioning (RI-CBT).

Design and Methods:

We retrospectively analyzed data of 155 patients who underwent RI-CBT performed at Toranomon Hospital from January 2005 to December 2008. The patients include 45 ABO-identical, 47 minor, 43 major, and 20 bidirectional ABO mismatched. All patients were performed using fludarabine-based reduced-intensity conditioning, and 114 patients (74%) were performed using TBI-containing regimen. Median age were 57 years-old and 94 patients (61%) were over 55 years-old. We evaluated the association between ABO incompatibility and neutrophil engraftment, one-year overall survival (OS) ; one-year non-relapsed mortality (NRM) ; and one-year relapse rate. We also analyzed the incidence of pre-engraftment immune reaction (PIR), acute graft-versus-host disease (GvHD) including severity, and thrombotic microangiopathy (TMA).

Results:

There were no significant differences in neutrophil-engraftment time, reticulocyte-engraftment time, and the incidence of PIR. The incidence of acute GvHD and grade 2-4 acute GvHD were significantly higher in major/bidirectional ABO-incompatible group than ABO-identical/minor ABO-incompatible group (respectively P=0.0008 and P=0.0116). The incidence of TMA tended to be higher in minor/bidirectional ABO-incompatible group than ABO-identical/major ABO-incompatible group (P=0.0637). There were no significant differences in one-year OS, NRM, and relapse rate. In multivariate analysis, risk factors of acute GvHD were age over 55, TBI-containing regimen, CD34-positive cells>0.7×10e5/kg, and major/bi-directional ABO incompatibility, and those of TMA were grade 3-4 acute GvHD and minor/bi-directional ABO incompatibility.

Discussion:

This study showed major-directional ABO incompatibility setting increased the incidence of acute GvHD. Sex incompatibility and HLA incompatibility were not significantly influenced the incidence of acute GvHD. The use of steroid for severe GvHD and the expression of ABO antigen on the surface of vascular endothelial cell may influence pathogenesis of TMA. Further studies including larger patients numbers are required to elucidate the impact of ABO incompatibility on the clinical outcome of RI-CBT.

Disclosures:

No relevant conflicts of interest to declare.
Higher Incidence of Relapse with Higher Doses of CD34+ Cells From Leukapheresis Products Infused in Adults with Acute Myelocytic Leukemia Autografted During the First Remission.
Authors: Gorin Norbert C , Labopin Myriam , Rocha Vanderson
Published: October 2019

Abstract 2315

Poster Board II-292

Purpose

The cell source for autologous stem cell transplantation has shifted from bone marrow (BM) to peripheral blood (PB). For patients with AML in CR1, we previously showed that the risk of relapse was greater with PB than BM and a poorer outcome was associated with a shorter interval from CR1 to PB transplantation (≦80 days) (JCO 2009 in press).

Leukemic and normal progenitors bear the CD34+ antigen and can be mobilized together; we questioned whether there was a relation linking the doses of CD34+ cells infused to the outcome.

Methods

Out of 1262 patients autografted with PB more than 80 days post CR1 and reported to the EBMT registry using MEDB form, the dose of CD34+ cells infused was available in 772.

Results

The CD34+ cell doses were categorized by percentiles to divide the whole group into five categories containing the same number of patients . We identified the fifth percentile (> 7.16 × 106/kg) as the cut off point for Relapse Incidence (RI) and Leukemia-Free survival (LFS). Patients receiving the highest dose had a higher RI (57+/-4%, vs. 44+/-2%; p=0.008) and a lower LFS (34+/-4% vs. 49+/-2%; p=0.007).

In a multivariate analysis adjusted for differences, RI was higher in patients receiving the highest CD34+ cell dose ((hazard ratio [HR], 1.48; 95% CI, 1.12-1.95; p = 0.005).and the LFS was worse (HR, 0.72; 95% CI, 0.55-0.93; p = 0.01).

Conclusion

For patients with AML in CR1 autografted with PB, risk of relapse is greater and LFS is lower in those receiving the highest doses of CD34+ cells.

Disclosures:

No relevant conflicts of interest to declare.
Pulmonary Infiltration by CLL/SLL in the Presence of Inflammation: Significant or Incidental Finding?.
Authors: Weil Andrew C , Kalaycio Matt E , Cook James R.
Published: October 2019

Abstract 2352

Poster Board II-329

Radiographically identified pulmonary infiltrates in patients with CLL are usually secondary to infectious causes. However, a subset of patients develop pathologic leukemic infiltrates. Lung biopsies containing both inflammation and CLL infiltrates are particularly problematic. It is unclear in such cases whether the CLL infiltrate represents a nonspecific “passenger effect” secondary to ongoing inflammation versus a pathologic leukemic infiltrate. Furthermore, it is unknown whether there are clinical factors that would help predict who is at higher risk for pulmonary infiltration by CLL. We retrospectively analyzed 45 transbronchial lung biopsies taken from 37 patients at the Cleveland Clinic between 1999 and 2008. We reviewed the biopsies for evidence of acute or chronic inflammation and involvment by CLL. Acute inflammation in our biopsies consisted of acute pneumonias. Cases displaying a lymphocytic infiltrate suspicious for CLL on routine H&E sections were further investigated by immunohistochemistry. The patients studied were 67.5% male with a median age of 67 (45-83) and a median absolute lymphocyte count of 2.75 (0.12 – 86.08). All patients at the time of diagnosis with CLL had flow cytometery of blood and/or bone marrow demonstrating CD5, CD19 positive B cells consistent with CLL. 72% of all patients were previously treated and 52% of patients had a RAI stage > 2 at the time of their diagnosis with CLL. CLL was identified in 20% of the biopsies. Of the 31.1% of biopsies showing acute inflammation, none had leukemia. CLL infiltration was found in association with and without chronic inflammation in 25% and 30.4% of the biopsies respectively. The median peripheral blood absolute lymphocyte count was 3.64 (1.33 – 86.08) in those with leukemic biopsies and 8.31 (0.12 – 79.12) in those without leukemic biopsies. Furthermore, only 33% of the patients with CLL positive biopsies had a RAI stage ≥ 3. At the time of biopsy, 7 previously treated patients were in remission; none of these cases displayed evidence of CLL in lung tissue. We found that the incidence of leukemic infiltration of lung biopsies did not increase in the presence of inflammation. In fact, the incidence of infiltration of biopsies with inflammation was lower than those without inflammation. These results demonstrate that leukemic infiltration on biopsies with inflammation is uncommon, and suggests that this phenomenon represents true, pathologic infiltration of the tissue by CLL and not a “passenger effect”. To the authors knowledge this is the largest study evaluating the association of inflammation and pathologic infiltration of pulmonary tissue by CLL.

Disclosures:

No relevant conflicts of interest to declare.
STAT3 Controls the Neutrophil Migratory Response to CXCR2 Ligands by Direct Activation of G-CSF-Responsive CXCR2 Expression and Via Modulation of CXCR2 Signal Transduction .
Authors: Nguyen-Jackson Hoainam , Panopoulos Athanasia , Zhang Huiyuan , Li Haiyan S. , Watowich Stephanie
Published: October 2019

Abstract 236

Neutrophil mobilization, the release of neutrophils from the bone marrow reserve into circulating blood, is important to increase peripheral neutrophil amounts during bacterial infections. Granulocyte colony-stimulating factor (G-CSF) and chemokines, such as MIP-2 (CXCL2), can induce neutrophil mobilization, but the mechanism(s) they employ remain unclear. The transcription factor STAT3 is the principal intracellular signaling molecule activated upon G-CSF-ligation of its receptor. Using a murine model with conditional STAT3 deletion in bone marrow, we demonstrated previously that acute G-CSF-responsive neutrophil mobilization and MIP-2-dependent neutrophil chemotaxis is mediated by STAT3. In this study, we show that STAT3 is also necessary for MIP-2-elicited neutrophil mobilization. STAT3 appears to function by regulating the amplitude of Raf/MEK/Erk activation by MIP-2, a signaling pathway that is important for MIP-2-mediated chemotaxis. In addition, we demonstrate that G-CSF stimulates the expression of the MIP-2 receptor via STAT3-dependent transcriptional activation of Il8rb. G-CSF treatment also induces changes in bone marrow chemokine expression levels in a STAT3-dependent manner, which may further affect neutrophil retention and release. Taken together, our study demonstrates that STAT3 regulates multiple aspects of chemokine and chemokine receptor expression and function within the bone marrow, indicating a central role in the neutrophil mobilization response.

Disclosures:

Nguyen-Jackson: Amgen, Inc: Research Funding. Panopoulos:Amgen, Inc: Research Funding. Zhang:Amgen, Inc: Research Funding. Li:Amgen, Inc: Research Funding. Watowich:Amgen, Inc: Research Funding.
Passive Transfer of Hepatitis B Core Antibody by Intravenous Immune Globulin in Patients with Immune Thrombocytopenic Purpura.
Authors: Arnold Donald M. , Carruthers Julie , DiTomasso Julie , Gagliardi Thea , Meyer Ralph M. , Crowther Mark A. , Kelton John G.
Published: October 2019

Abstract 2418

Poster Board II-395

Background:

Rituximab may reactivate remote hepatitis B infection. Thus, hepatitis B core antibody (HBcAb) seropositivity was an exclusion for a randomized trial of rituximab in patients with immune thrombocytopenic purpura (ITP) in Canada. During screening for this trial, we observed a high rate of HBcAb seropositivity at our centre. We investigated passive antibody transfer by intravenous immune globulin (IVIg) as a potential explanation.

Methods:

We performed a case-control study of consecutive ITP patients screened for eligibility for a randomized trial of rituximab. We compared the frequency of prior IVIg use and the IVIg product administered to patients who tested positive (n=11) and negative (n=13) for HBcAb. Total (IgG and IgM) HBcAb was tested using a microparticle enzyme immunoassay (MEIA) that measures the rate of inhibition compared with an internal cut-off value (AxSYM®, Abbott Laboratories, IL, USA). We calculated the odds ratio (OR) and 95% confidence interval (CI) for IVIg exposure. We also tested different lots of 2 IVIg products directly for HBcAb in the MEIA.

Results:

Of 24 consecutive patients with ITP screened, 11 (45.8%) were positive for HBcAb and thus were excluded from the randomized trial. The observed rate of HBcAb seropositivity in this cohort was 35 times higher than the expected seroprevalence in Canada of 1.3% (O'Brien Transfusion 2009). Of 11 HBcAb-positive patients, 10 (90.9%) had received prior IVIg [OR, 16 (95% CI, 1.54, 166.05)]. Of the 10 patients who received IVIg, 8 (80.0%) received Gamunex (10% human immune globulin, Talecris Biotherapeutics, Clayton, NC). HBcAb was repeated in 8 seropositive patients, of whom 4 were negative on repeat testing 7 to 104 weeks after the last dose of IVIg. Of the 13 HBcAb-negative patients, 5 (38.5%) had received prior IVIg; none were Gamunex. Seroconversion was documented in one patient serially tested after receiving Gamunex; MEIA rates decreased (became positive) rapidly, then gradually increased to normal after 7 weeks. Samples from 4 lots of Gamunex were reactive in the MEIA; whereas 3 lots of IGIVnex (Talecris Biotherapeutics) were non-reactive.

Conclusions:

False positive HBcAb results may occur due to the passive transfer of HBcAb from Gamunex, an IVIg product manufactured from donors who are not screened for HBcAb. HBcAb should be interpreted cautiously in any patient who received IVIg. We recommend HBcAb testing before IVIg administration so that treatments such as rituximab are not withheld unnecessarily.

Disclosures:

Arnold: Hoffman-LaRoche: Research Funding; Amgen: Consultancy, Honoraria. Meyer:The NCIC CTG is a clinical trials cooperative group. It receives its base grant support from the Canadian Cancer Society Research Institute. Many of its clinical trials include research support from industry. : Research Funding.
The Effects of Hyperthermia On Platelet Physiology.
Authors: Shi Quanwei , Wang Zhicheng , Yan Rong , Du Juan , Liu Jun , Ruan Changgeng , Dai Kesheng
Published: October 2019

Abstract 2422

Poster Board II-399

Introduction:

In some patients with fever or hyperthermia, hemorrhage is a significant pathological feature, which incurs severe or even fatal consequence. Although the mechanisms of hemorrhage in patients with hyperthermia or fever have been though to be complex, whether there is an association between hemorrhage and hyperthermia or fever is not well understood. Platelets play a central role in maintaining integrity of endothelium and biological hemostasis. Platelet count obviously reduces in most of patients with dengue fever or heatstroke, and reduced platelet aggregations have been found in patients with hemorrhage fever, raising the possibility that fever or hyperthermia incurs reduction of platelet count or function leading to hemorrhage.

Methods and Results:

To explore the effect of hyperthermia on platelet function, platelet-rich plasma (PRP) was isolated and incubated at hypothermia (22 degrees C), normothermia (37 degrees C) or hyperthermia (40 and 42 degrees C) for 1 or 2 hours, and then induced to aggregation by ADP. Platelet aggregation was significantly reduced with the growth of temperature in a time-dependent manner. To exclude the possible interference from plasma proteins, and to further investigate the effects of hyperthermia on platelet function, alpha-thrombin induced platelet aggregations were examined in washed platelets incubated at different temperatures for 1 or 2 hours. Washed platelets presented normal aggregation response to alpha-thrombin in platelets incubated at hypothermia, whereas alpha-thrombin induced platelet aggregations reduced gradually with the increase of temperature and time duration. Next, we investigated whether the reductions of platelet aggregation were resulted from platelet apoptosis under high temperature conditions. Hyperthermia gradually induced apoptotic events in platelets with the increase of temperature and time duration, including depolarization of mitochondrial inner transmembrane potential (ΔΨm), gelsolin cleavage, and phosphatidylserine (PS) exposure. Furthermore, hyperthermia incurs platelet glycoprotein (GP) Ibalpha ectodomain shedding.

Conclusions:

These results indicate that hyperthermia induces platelet apoptosis, thus suggesting the possible reason why platelet count or function was reduced in some patients with fever or hyperthermia. These findings not only have important implications for the pathogenesis of hemorrhage in some fever or hyperthermia-related diseases, but also suggest that attentions should be paid on platelet apoptosis under relative high temperature conditions, such as during hyperthermia therapy or platelet storage.

Disclosures:

No relevant conflicts of interest to declare.
Health Outcomes and Costs of Rituximab in Combination with Cyclophosphamide, Vincristine and Prednisolone in the Treatment of Patients with Advanced Follicular Lymphoma in Portugal.
Authors: Braga Paula , Carvalho Susana , Gomes Marília , Guerra Lurdes , Lúcio Paulo , Marques Herlander , Negreiro Filipa , Pereira Catarina , Silva Catarina , Teixeira Adriana
Published: October 2019

Abstract 2481

Poster Board II-458

Objective:

With the pressure on healthcare budgets, it has become increasingly important for healthcare decision makers to consider the value for money of the treatments they reimburse. The objective of this analysis was to evaluate the long term outcomes and costs of rituximab in combination with cyclophosphamide/vincristine/prednisolone chemotherapy regimen (R-CVP) versus CVP alone, in previously untreated patients with indolent Non-Hodgkin Lymphoma (NHL) from the Portuguese National Health System (NHS) perspective.

Methods:

Cost-effectiveness (Life Years Gained - LYG) and cost-utility analysis (Quality Adjusted Life years – QALYs) were performed for a time horizon of 10 years, according to a Markov economic model with three health states (“progression free survival”, “progression” and “death”) and monthly cycles for a population of previously untreated patients with indolent NHL. Data from a phase III clinical trial (Marcus 2007) was used and expanded to include unpublished 53-month median follow-up data. Survival after first-line therapy was estimated from the Scotland and Newcastle Lymphoma Group registry data and utilities were derived from a study in the UK performed in patients with follicular lymphoma. Resource consumption was estimated by a Portuguese expert panel (Delbecq Panel). Costs were calculated from the Portuguese NHS perspective through official data with prices updated to 2008. Only direct medical costs were considered. Costs and clinical outcomes were discounted at 5% per annum. Deterministic and probabilistic sensitivity analysis were performed around assumptions on the time horizon, costs, utilities and excess mortality rate due to progression applied in the base-case analysis.

Results:

The 10-year base-case analysis showed a lower total cost per patient with CVP alone (€ 85,838) in comparison with R-CVP (€ 87,774). Life expectancy and quality-adjusted life expectancy per patient were higher with R-CVP (6.361 and 4.166, respectively) than with CVP alone (5.557 and 3.438, respectively), representing increases of 0.804 in LYG and 0.728 (8.7 months) in QALYs gained. The incremental cost per LYG was € 2,407 and the incremental cost per QALY gained was € 2,661. The probabilistic sensitivity analysis confirmed the robustness of the base-case analysis results.

Conclusions:

This study demonstrates that the combination R-CVP in previously untreated non-Hodgkin lymphoma patients improves life expectancy and is a cost-effective alternative to CVP in Portugal.

Disclosures:

Braga: Roche Farmacêutica Química, Lda: Employment. Pereira:Roche Farmacêutica Química, Lda.: Employment.
Quality of Life in Non-Hodgkin Lymphoma: A Comparison Between Survivors of Indolent and Aggressive Disease.
Authors: Blaes Anne Hudson , Ma Linan , Peterson Bruce A.
Published: October 2019

Abstract 2494

Poster Board II-471

Introduction:

Few studies have examined the functional limitations, physical health, mental health, and general quality of life in cancer survivors with various types of non-Hodgkin lymphoma (NHL). Patients with aggressive NHL (AGG) require immediate chemotherapy for a potentially curable disease in contrast to patients with indolent NHL (IND) that is characterized by a chronic course with repeated relapses and progression despite therapy. This study examined the QOL in adult survivors of NHL >1 year from diagnosis and not currently on therapy; we hypothesized that fatigue and quality of life would be worse in patients with low grade NHL given the expectation of future progressive disease and repetition of multiple therapies.

Methods:

109 patients with NHL (58 AGG, 51 IND), more than one year from initial diagnosis, and at least three months from any active therapy, completed two health related quality of life assessments using the Medical Outcomes Study 36-Item Short-Form Healthy Survey (MOS SF-36) which assesses 8 subsets and 2 summary scores, and the Functional Assessment in Cancer Therapy – Fatigue (FACT-F). Physical and Mental SF-36 scores or FACT-F scores between IND and AGG were compared using a two sample t-test. Multiple linear regression* was performed to account for any potentially explanatory variables (age, use of chemotherapy, time from last treatment to survey).

Results:

Median age was 60 years [61 years (33-88) IND and 57 years (22-90) AGG]. 48.6% were female (52.9% IND, 44.8% AGG). 74.3% were in complete remission at the time of the survey (52.9% IND, 93.1% AGG) (p<0.001). 70.6% had received chemotherapy (43.1% IND, 94.8% AGG) and 55% had received immunotherapy (31.4% IND, 75.9% AGG). 17.6% of IND had received no therapy. The overall physical (PCS) and mental (MCS) component quality of life scores of the SF-36 did not differ between survivors with aggressive and indolent NHL; the median PCS was 51.8 (11.2-66.5) [56.8 IND, 51.4 AGG, p=0.192] and the median MCS was 54.2 (10.8-65.8) [54.6 IND, 53.3 AGG, p=0.239]. Examining the eight subcategories of the SF-36, physical function in survivors of IND was significantly better when compared with those of AGG NHL. Using the FACT-F, fatigue scores also did not differ between the two populations; the median FACT-F score was 45 (8-52) [47 IND, 43 AGG, p=0.114).

Conclusion:

While some other studies have reported a decreased quality of life in NHL survivors, our study reports a similar overall quality of life between survivors of IND and AGG NHL more than one year from initial diagnosis; this may be explained by a significant proportion of our IND patients having never received therapy or an acclimation to the need for future therapy. Physical function may be more impaired in survivors of AGG NHL and warrants further investigation.

Disclosures:

No relevant conflicts of interest to declare.