bluuebunny/crossref_metadata_2025 huggingface dataset:
Hospitalization Outcome Metrics Based On Payer Status In Myeloma Patients That Receive Autologous Stem Cell Transplant (ASCT)
Authors: Gowda Sachit , Langston Amelia , Kaufman Jonathan L. , Shah Nishi N , Lechowicz Mary Jo , Boise Lawrence H. , Lonial Sagar , Nooka Ajay K.
Published: October 2019

Background

Myeloma patients have experienced great survival benefits in the last decade due to the use of novel agents and autologous stem cell transplant (ASCT). Prior studies report a complex interplay between payer status and the receipt of ASCT. We have evaluated if the payer status affects outcome metrics of length of stay (LOS), in-hospital mortality rate (IHM),  and total hospitalization charges in the context of survival benefit of myeloma with this procedure.

Methods

We used the NIS (Nationwide Inpatient Sample) 2001-2010 database (part of the HCUP database) to obtain the patient data.  Using private insurance as the reference group, we performed multivariate logistic regression to understand the association of payer status with LOS, IHM, hospitalization charges. We adjusted our model for age, race and the presence or absence of co-morbidities. Comorbidites were identified using comorbidity software that created measures reported by Elixhauser et al.

Results

From 01/2001 until 12/2010, 25656 admissions for ASCT as principal procedure for the principal diagnosis of multiple myeloma were included in our analysis. The IHM rate during this period based on payer status was 3.04%, 1.56%, 1.20% and 0.4% for medicare, medicaid, private insurance and others, respectively. Median LOS for medicaid and medicare were 17 days while private insurance and other insurances had a median LOS of 16 days. Medicare patients undergoing ASCT had higher likelihood of IHM compared to private insurance [Odds ratio: 2.62 ( 95%CI 1.46 – 4.72)]; while medicaid patients had non-significant increase. LOS in medicaid patients was longer compared to private insurance [(Odds ratio: 1.53 ( 95%CI 1.16-2.02)].

Conclusion

 Myeloma patients with medicare undergoing ASCT had higher likelihood of in-hospital death compared to patients with private insurance. Medicaid patients had a lengthier in-hospital stay but there seems to be no significant difference in hospitalization charges in the different payer groups. However, the acceptable overall cumulative mortality rate suggests that myeloma patients can continue to enjoy the survival benefits associated with ASCT despite payer status. Further studies evaluating long-term outcomes outside the hospital admission would be required to better understand the association of payer status with overall survival benefits of ASCT.

Disclosures:

Kaufman: Onyx: Consultancy; Celgene: Consultancy, Research Funding; Novartis: Consultancy, Research Funding; Janssen: Consultancy; Millenium: Consultancy; Merck: Research Funding. Boise:Onyx: Consultancy. Lonial:Millennium: Consultancy; Celgene: Consultancy; Novartis: Consultancy; BMS: Consultancy; Sanofi: Consultancy; Onyx: Consultancy.
Upfront Allogeneic-Stem Cell Transplantation for Patients with Non-Localized Untreated Peripheral T-Cell Lymphoma: An Intention-to-Treat Analysis from a Single Center
Authors: Loirat Marion , Chevalier Patrice , Leux Christophe , Moreau Anne , Bossard Céline , Guillaume Thierry , Gastinne Thomas , Delaunay Jacques , Blin Nicolas , Mahe Beatrice , Dubruille Viviane , Augel-Meunier Karine , Peterlin Pierre , Maisonneuve Herve , Morineau Nadine , Moreau Philippe , Jardel Henry , Mohty Mohamad , Moreau Philippe , Legouill Steven
Published: October 2019

Background. Peripheral T-cell lymphomas (PTCLs) are rare and heterogeneous diseases with dismal outcome when treated with chemotherapy alone. Because allogeneic stem cell transplantation (allo-SCT) can cure relapse-refractory patients, we hypothesised that upfront allo-SCT may be less toxic and provide a better outcome. Therefore, all patients that presented with advanced PTCL in our institution at diagnosis were scheduled to undergo upfront allo-SCT after induction chemotherapy.

Patients and methods. The aim of the present work was to assess the feasibility and toxicity of upfront allo-SCT. From 2004 to 2012, 49 newly diagnosed PTCL patients were scheduled to receive upfront allo-SCT. An HLA-matched donor was found for 42 patients: related to the patient in 15 cases, unrelated in 20 cases, and suitable cord blood units were used in seven cases.

Results. After induction chemotherapy, 17 patients reached complete remission and 29 (60%) proceeded to upfront allo-SCT. For all patients, the one and two-year overall survival (OS) rates were 59% (CI95%; 47-75) and 55 % (CI95%; 43-71), respectively. The most frequent reason we did not proceed to allo-SCT was disease progression or insufficient response after induction. For transplanted patients, the one- and two-year OS were 76% (CI95%; 62-93) and 72.5% (CI95%; 58-91), respectively. Toxicity related mortality one year after allo-SCT was only 8.2% (CI95%; 0-18.5). The two-year progression-free survival (PFS) rate of patients who did not proceed to allo-SCT (n= 20) was below 30%. The disease status at the time of transplantation was a strong predictive marker for both PFS and OS in transplant patients.

Conclusions. Upfront allo-SCT in PTCLs is feasible with low TRM, and it provides long-term disease control. However one third of patients remain chemo-refractory, and thus, new therapeutic approaches are warranted. The role of upfront allo-SCT compared to other therapeutic approaches in PTCLs requires investigation in randomized studies.

Disclosures

No relevant conflicts of interest to declare.
SEC23B Deficiency Results in Different Phenotypes in Humans and Mice
Authors: Khoriaty Rami , Vasievich Matthew , Jones Morgan , Chase Jennifer , Everett Lesley , Zhang Bin , Maillard Ivan , Ginsburg David
Published: October 2019

SEC23B mutations in humans result in the autosomal recessive disease Congenital Dyserythropoietic Anemia type-II (CDAII). CDAII is characterized by moderate anemia in increased bone marrow (BM) bi/multi-nucleated erythroblasts. Despite the identification of the genetic defect underlying CDAII, the pathophysiology of this disease remains unknown. SEC23A and SEC23B are paralogous components of the coat protein complex II (COPII)-coated vesicles, which transport secretory proteins from the Endoplasmic Reticulum to the Golgi apparatus. We generated SEC23B-deficient mice and demonstrated that these animals die perinatally exhibiting massive pancreatic degeneration (Tao et. al PNAS). To examine the impact of SEC23B-deficiency on adult murine hematopoiesis, we harvested fetal liver cells (FLC), which contain hematopoietic stem cells, from SEC23B-deficient or wild-type (WT) control E17.5 embryos and transplanted them into lethally irradiated C57BL/6J mice. Recipients of SEC23B-deficient FLC did not exhibit anemia or any other CDAII characteristic (Khoriaty et. al, Mol Cell Biol), and SEC23B deficient FLC competed effectively with WT FLC at reconstituting hematopoiesis when transplanted into lethally irradiated recipient mice (Khoriaty et. al, Mol Cell Biol). A Sec23bfl conditional-allele was also generated. Mice with hematopoietic-specific SEC23B-deficiency, generated by crossing Vav1-Cre into Sec23bfl mice, also exhibited normal hematopoiesis. In contrast, mice with pancreas-specific Sec23b deficiency generated by crossing the Sec23bfl allele to a p48-Cre or Pdx1-Cre resulted in a phenotype indistinguishable from complete SEC23B-deficiency, demonstrating that loss of pancreatic Sec23b expression is sufficient to explain the perinatal lethality of SEC23B-deficient mice. To investigate different phenotypes of SEC23B deficiency in humans and mice, the SEC23B/SEC23A expression ratio was examined in murine and human tissues. This ratio is higher in mouse pancreas (12.7) compared to BM (2.6), whereas it is higher in human BM (7.8) relative to pancreas (5.5). Taken together with the high degree of amino-acid identity between SEC23A and SEC23B (~85%), these data suggest that the tissue-specific functions of SEC23A and SEC23B have shifted during evolution between humans and mice. To determine if Sec23a can rescue the lethality of SEC23B-deficient mice, we have engineered Sec23a cDNA into the endogenous genomic locus of Sec23b (Sec23A-B) via recombinase-mediated cassette exchange. A heterozygous Sec23A-B intercross is in progress. Rescue of the SEC23B deficient phenotype by SEC23A protein expressed under control of Sec23b regulatory sequences would suggest that increasing the expression of either paralog in erythroid cells might be effective in the treatment of CDAII.

Disclosures

No relevant conflicts of interest to declare.
Single Cell Transcriptome-Based Dissection of Lineage Fate Decisions in Myelopoiesis
Authors: Olsson Andre , Grimes H. Leighton , Chaudhri Virendra K , Dexheimer Philip , Aronow Bruce J , Singh Harinder
Published: October 2019

In spite of tremendous advances in the analysis of hematopoietic progenitors and transcription factors that give rise to different lineages, molecular insight into the mechanisms that underlie cell fate choice at the level of individual cells is lacking. We utilized single-cell RNA sequencing of murine granulocyte-monocyte progenitors (GMPs) to analyze the molecular basis of cell fate choice. Over 200 libraries were generated with average read depths of 4 million per library and an expressed gene call of over 3,800 genes with FPKM >3. Our data reveal a varied but coherent spectrum of gene expression patterns in individual murine GMPs. The majority of cells could be clustered into ones expressing either granulocytic or monocytic genes, suggesting that they were primed for lineage determination. A minority of GMPs expressed a mixed-lineage pattern of genes. The single-cell data suggested an antagonistic transcription factor circuit involving Gfi1 and IRF8 that was validated with both loss- and gain-of-function experiments in GMPs. Our data highlight the utility of single cell RNA-Seq analysis to reveal molecular mechanisms controlling lineage fate decisions in hematopoiesis.

Disclosures

No relevant conflicts of interest to declare.
IL-21-Mediated Generation of Human Regulatory B Cells Occurs Independently of CD40 Signaling
Authors: Jahrsdörfer Bernd , Kaltenmeier Christof , Gawanbacht Ali , Beyer Thamara , Schütz Catharina , Hönig Manfred , Kirchhoff Frank , Schrezenmeier Hubert
Published: October 2019

Recently, we and others found that B cells differentiate into regulatory B cells (Breg) in response to interleukin (IL-)21. Of note, the key characteristic of human IL-21-induced Breg is expression of the serine protease granzyme B (GrB), whereas murine Breg, which require both IL-21 and CD40 ligand (CD40L) for their induction, predominantly express the immunosuppressive cytokine IL-10. Using two different disease models and various immunological methods, we further characterized the conditions leading to Breg differentiation in humans. Here, we demonstrate that in humans CD40L determines whether IL-21 induces differentiation of B cells into plasma cells (CD40L presence) or into GrB+ Breg (CD40L absence), which can directly control T cell proliferation by GrB-dependent degradation of the T cell receptor z-chain. Furthermore, we show that GrB+ Breg are circulating at high frequencies in the peripheral blood of untreated, highly viremic HIV patients, but not in healthy subjects. Of note, HIV-infected CD4+ T cells express IL-21, but not CD40L, and induce a GrB+ regulatory phenotype in healthy third party B cells in vitro. Consequently, addition of CD40L multimers can compensate for this insufficient T helper cell function, resulting in increased plasma cell/Breg ratios. Moreover, we investigated a patient with a congenital defect of Nuclear-Factor-kappa-B-Essential-Modulator (NEMO), which is essential for normal CD40 signaling. Even in the presence of viral infections, when CD4+ T helper cells from such patients are highly activated with strong expression of IL-21, they are not able to establish sufficient antibody responses. Instead, we found this patient to almost exclusively harbor B cells with a regulatory phenotype including high basal levels of GrB. When untreated NEMO B cells were co-cultured with allogeneic T cells from a healthy third party donor, these T cells failed to proliferate and to survive in response to a 6-day stimulation with anti-CD3/CD28 antibodies, an effect not observed with B cells from healthy donors. Since NEMO B cells lack normal CD40 signaling, our findings unequivocally demonstrate that in contrast to murine Breg IL-21-dependent induction of human Breg can occur in a CD40-independent fashion.

Disclosures

No relevant conflicts of interest to declare.
NADPH Oxidase Mediates Antiphospholipid Antibody-Induced Endothelial Cell Activation
Authors: Wu Meifang , McCrae Keith R.
Published: October 2019

Introduction: Antiphospholipid Syndrome (APS) is characterized by thrombosis and/or recurrent fetal loss in the presence of persistently elevated antiphospholipid antibodies (APLA). The majority of pathologic APLA are directed against β2-glycoprotein I (β2GPI), an abundant plasma phospholipid binding protein. APLA/anti-β2GPI antibodies activate endothelial cells in a β2GPI-dependent manner, though the underlying mechanisms are not well defined.

Objective: To define the role of NOX1 in the generation of ROS and activation of endothelial cells by anti-β2GPI antibodies.

Methods: Endothelial cells were incubated with β2GPI and either control or affinity-purified anti-β2GPI antibodies in the absence or presence of diphenyleneiodonium (DPI), an NADPH oxidase (NOX) inhibitor. Generation of reactive oxygen species (ROS) in treated cells and conditioned medium were measured by using fluorescent dyes (CM-H2DCFDA and CellROX Deep Red) or luminescent substrate. NOX mRNA and protein expression were assessed using quantitative PCR and immunoblot. Endothelial cell activation was measured by increased expression of E-selectin.

Results: Incubation of endothelial cells with β2GPI and anti-β2GPI antibodies stimulated ROS generation in endothelial cells, as well as the release of ROS into conditioned medium. The expression of NOX1 mRNA and protein levels were significantly increased in endothelial cells exposed to anti-β2GPI antibodies, but not control IgG (Figure 1). The ability of β2GPI and anti-β2GPI antibodies to induce endothelial cell E-selectin mRNA expression was blocked by pretreatment of cells with DPI (Figure 2), suggesting that ROS is required for downstream events underlying endothelial cell activation.

Conclusions: Endothelial cells exposed to β2GPI and anti-β2GPI antibodies generate ROS, which is subsequently released into the conditioned medium. NOX1 appears to be essential for ROS generation. The impairment of endothelial cell activation by DPI suggests that NOX is also essential for endothelial cell activation by anti-β2GPI antibodies. We hypothesize that production of ROS by NOX plays a central role in APLA-induced endothelial dysfunction.

Acknowledgment: This work was supported by an ASH Bridge Grant

Figure 1 Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Characterization of Patients with Glanzmann Thrombasthenia and Identification of 17 Novel Mutations
Authors: Sandrock-Lang Kirstin , Oldenburg Johannes , Wiegering Verena , Halimeh Susan , Santoso Sentot , Kurnik Karin , Fischer Lars , Tsakiris Dimitrios , Sigl-Kraetzig Michael , Brand Brigit , Kraetzer Katharina , Deeg Niklas , Busse Eileen , Barbara Zieger
Published: October 2019

Background: Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder. Platelets from patients with GT show quantitative and/or qualitative defects of the platelet membrane glycoprotein (GP) IIb/IIIa complex, also called integrin αIIbβ3. On activated platelets the αIIbβ3 binds von fibrinogen and Willebrand factor which leads to platelet spreading and formation of platelet-platelet protein bridges.

Patients: In this study, 18 patients with GT were investigated with molecular genetic analyses. The patients presented with bleeding symptoms such as epistaxis, mucocutaneous bleeding, haematomas, petechiae, gastrointestinal bleeding, and menorrhagia.

Methods and Results: As cause of GT in the patients homozygous or compound heterozygous mutations in ITGA2B or ITGB3 were identified through sequencing of genomic DNA. All exons including exon/intron boundaries of both genes were analyzed. In summary, 16 of 18 patients revealed 27 different mutations (ITGA2B: n = 17, ITGB3: n = 10). Of these mutations, 17 have not been published yet.

Conclusion: In 16 patients mutations in ITGA2B or ITGB3 were identified as cause of GT. A total of 27 mutations including 17 novel missense, nonsense, frameshift and splice site mutations were detected. None of these mutations were present more than once in unrelated patients. In addition, 2 patients were without molecular genetic findings in ITGA2B or ITGB3 that could explain the suspected diagnosis of GT. We hypothesize that these patients may harbour defects in a regulatory element affecting the transcription of these genes or there may exist other proteins important for the activation of the αIIbβ3 complex that could be affected.

Disclosures

No relevant conflicts of interest to declare.
Effects of the Timing of Venous Thromboembolic Events on Survival of Patients with Uterine Cancer
Authors: Remasankar Heera , Zhou Xiao , Vadhan-Raj Saroj
Published: October 2019

Background: Cancer patients are at high risk for venous thromboembolic event (VTE), and the occurrence of VTE can affect the prognosis. However, it is unclear if the timing of VTE can influence the prognosis. The purpose of this study was to evaluate the incidence and impact of the timing of VTE (early vs late) on the survival of patients with uterine cancers.

Methods: A retrospective cohort study was conducted. The study population included all uterine cancer patients who had newly diagnosed episode of VTE from January 1st 2006 to December 31st2007 at MD Anderson Cancer Center. Medical records of these patients were reviewed for cancer diagnosis, patient demographics, metastasis, date of diagnosis of VTE, type of VTE, the site of VTE, and any recurrences during the follow up period of 2 years since the first VTE. Clinical and laboratory parameters predictive for survival were also reviewed. All VTE episodes, including symptomatic as well as incidental VTEs were confirmed by the radiological studies using CT ANGIO, CT scan, Doppler compression ultrasound or V/Q perfusion scans. The survival time was defined as the time from the date of cancer diagnosis to the date of death, or to the date of last follow up. Survival analysis was conducted using Kaplan-Meier method and Cox proportional hazard models.

Results: Of the 2151 patients with newly diagnosed VTEs within the 2 years study period, 33 patients were found to have uterine cancers. The median age was 60 years (range 41-84 years). Among all the primary VTEs, 42% were pulmonary embolus (PE); 45% were deep vein thrombosis (DVT), and 12% were concurrent PE/DVT (diagnosed on the same day). The median time of VTE from the date of cancer diagnosis was 8 months (range 0 - 88 months). The majority of these VTE events (55%) were found to occur within 1 year from the date of cancer diagnosis. A total of 8 patients had 10 recurrent episodes of VTE (2 patients had 2 recurrent VTE episodes) during the follow up period. The median survival time was 33 months (range 2.6-121.6 months); 82% of these patients had a metastatic disease. Multivariate proportional hazards model showed that the diagnosis of VTE within 6 months from the cancer diagnosis [Hazard ratio: 3.8, 95% CI (1.1-12.9), p=0.03] and the presence of baseline white blood cell (WBC) count of greater than 11,000/uL [Hazard ratio: 3.5, 95% CI (1.0-12.2), p=0.045] were statistically significant independent predictors for 2 year survival adjusting for age, presence of central venous catheter (CVC), and the baseline platelet count of greater than 350,000/uL.

Conclusion: These findings suggest that the timing of VTE is an important indicator of prognosis in patients with uterine cancer; patients who had VTE within 6 months from cancer diagnosis had a shorter 2 year survival. Future larger studies may better define the impact of timing of VTE on survival of patients with Uterine Cancers.

Disclosures

No relevant conflicts of interest to declare.
Critical Role of T Cells in PF4/Heparin Antibody Production
Authors: Zheng Yongwei , Yu Mei , Padmanabhan Anand , Aster Richard H. , Wen Renren , Wang Demin
Published: October 2019

Heparin-induced thrombocytopenia (HIT) is an antibody-mediated disorder that can cause arterial or venous thrombosis/thromboembolism, and platelet factor 4 (PF4)/ heparin-reactive antibodies are essential to the pathogenesis of HIT. Our recent studies have demonstrated that marginal zone (MZ) B cells play a major role in production of PF4/heparin-specific antibodies. However, the role of T cells in production of these pathogenic antibodies is not clear. Here we showed that PF4/heparin complex-induced production of PF4/heparin-specific antibodies was markedly impaired in mice, in which CD4 T cells were depleted by administration of GK1.5 anti-CD4 monoclonal antibody. As expected, the CD4 T cell-depleted mice responded normally to T cell-independent antigen TNP-Ficoll but not T cell-dependent antigen NP-CGG, in agreement with the lack of CD4 T cells in these GK1.5-treated mice. Further, following adoptive transfer of a mixture of wild-type splenic B cells and splenocytes from B cell-deficient μMT mice, T and B cell-deficient Rag1 knockout mice responded to PF4/heparin complex challenge to produce PF4/heparin-specific antibodies. In contrast, Rag1-deficient mice that received a mixture of wild-type splenic B cells and splenocytes from Rag1-deficient mice barely produced PF4/heparin-specific antibodies upon PF4/heparin complex challenge. These data suggest that T cells are required for production of PF4/heparin-specific antibodies. Consistent with this concept, mice with B cells lacking CD40 molecule, a B cell costimulatory molecule that helps T cell-dependent B cell responses, displayed a marked reduction of PF4/heparin-specific antibody production following PF4/heparin complex challenge. Also as expected, mice with CD40-deficient B cells were able to respond to T cell-independent antigen TNP-Ficoll but not T cell-dependent antigen NP-CGG, consistent with the lack of T-cell help in these mice. Taken together, these findings demonstrate that T cells play an essential role in production of PF4/heparin-specific antibodies by MZ B cells.

Disclosures

No relevant conflicts of interest to declare.
Histone Acetylation Mediated by Brd1 Is Crucial for Cd8 Gene Activation during Early Thymocyte Development
Authors: Mishima Yuta , Wang Changshan , Miyagi Satoru , Saraya Atsunori , Hosokawa Hiroyuki , Yamaguchi Naoto , Nakayama Toshinori , Koseki Haruhiko , Taniuchi Ichiro , Iwama Atsushi
Published: October 2019

During T cell development, Cd8 expression is controlled via dynamic regulation of its cis-regulatory enhancer elements. Insufficient enhancer activity during initial Cd8 activation is known to cause a variegated CD8 expression, thereby generating a CD4+CD8-TCRb-/low population that should appear as CD4+CD8+ double-positive (DP) thymocytes. However, epigenetic and molecular mechanisms involved in initial Cd8 gene activation remain elusive. We previously reported that Brd1, also known as Brpf2, is responsible for the global acetylation of H3K14 as a subunit of the Hbo1 histone acetyltransferase (HAT) complex.

In this study, we generated conditional Brd1 knockout mice (Tie2-Cre;Brd1fl/fl mice), in which Brd1 is inactivated in all hematopoietic cells. Although Tie2-Cre;Brd1fl/fl mice were born and grew normally, detailed analysis revealed an abnormal thymocyte differentiation. Deletion of Brd1 resulted in the appearance of CD4+CD8-TCRβ-/low thymocytes that was indistinguishable from DP thymocytes in their properties. Hierarchical clustering of gene expression profile showed that Brd1Δ/Δ CD4+CD8-TCRβ-/low thymocytes retain an expression profile nearly identical to one observed in DP thymocytes. These results indicate that Brd1Δ/Δ CD4+CD8-TCRβ-/low thymocytes correspond to immature thymocytes that would normally appear as CD4+CD8+ DP thymocytes, and was generated because of a variegated activation of Cd8 gene. Importantly, in the gut intraepithelial lymphocyte (IEL) compartment, the population of CD4+CD8aa+ aβT cells, whose generation requires Cd8 gene reactivation in CD4+CD8− helper T cells, were significantly decreased in Tie2-Cre;Brd1fl/fl mice compared to the Tie2-Cre control mice. In addition, the percentage of CD8aa expressing gdT cells was significantly reduced. These findings suggest that Brd1 is also required to initiate Cd8a activation in gdT cells as well as Cd8 reactivation in CD4+T cells.

We further conducted retroviral transduction of Brd1 into Brd1Δ/Δ DN3 cells that were flowed by culture on TSt-4/DLL stromal cells. Upon exogenous Brd1 expression, CD8 expression was completely restored and thereby Brd1Δ/Δ DN3 cells differentiated into DP cells.

ChIP analysis demonstrated that Brd1 and Hbo1 co-localize at the known enhancers in the Cd8 locus and that their bindings are responsible for acetylation at H3K14. Biochemical results confirmed a presence of Brd1 in Hbo1 HAT complexes. These findings indicate that the Brd1-mediated HAT activity is crucial for efficient activation of Cd8 expression via acetylation at H3K14, which would serve as an epigenetic mark that promotes the recruitment of transcription machineries to the Cd8 enhancers.

Disclosures

No relevant conflicts of interest to declare.
Ripk-Mediated Necroptosis Induces Inflammation and Bone Marrow Failure in Mice
Authors: Roderick Justine E. , Hermance Nicole , Zelic Matija , Simmons Matthew , Polykratis Apostolos , Pasparakis Manolis , Kelliher Michelle
Published: October 2019

TNF-α and IFN-γ overproduction are features associated with human bone marrow failure syndromes such as Fanconi Anemia (FA) and Aplastic Anemia (AA). Cells from these patients are known to be hypersensitive to TNF-α and IFN-γ-induced cell death. The serine threonine kinases RIPK1 and RIPK3 interact to mediate necroptosis induced by TNF-α, type I or II interferons. We demonstrate that a hematopoietic RIPK1 deficiency results in hematopoietic stem and progenitor cell loss and induction of bone marrow failure. The cell death reflects cell-intrinsic survival roles for RIPK1 in hematopoietic stem and progenitor cells, as Vav-iCre Ripk1fl/fl fetal liver cells failed to reconstitute hematopoiesis in lethally irradiated recipients. Hematopoietic failure in these mice is accompanied by increases in serum pro-inflammatory cytokines/chemokines and reduced hematopoietic colony formation in the presence of TNF-α, type I or II interferon. We provide genetic evidence that a RIPK3 deficiency rescues the bone marrow failure and significantly reduces serum cytokine and chemokine levels in Vav-iCre Ripk1fl/fl mice. These data reveal that in the hematopoietic lineage RIPK1 prevents inflammation by suppressing RIPK3 activity and raise the possibility that human bone marrow failure patients may benefit from selective RIPK inhibitors.

Disclosures

No relevant conflicts of interest to declare.
Temozolomide Induced Bone Marrow Suppression- a Single Institution Outcome Analysis and Review of Literature
Authors: Kourelis Taxiarchis , Buckner Jan C , Gangat Naseema , Patnaik Mrinal M.
Published: October 2019

Background: Severe myelosuppression (MS) and bone marrow failure (BMF) are rare but well characterized adverse effect of temozolomide (TMZ). The existing literature is limited to case reports and data reported to regulatory agencies, which usually describe very severe cases and lack long term and detailed follow-up.

Methods: We identified cases of myelosuppression attributed to TMZ seen at the Mayo clinic, from 1998-2014. We also performed a literature (PubMed) search to identify published cases of TMZ induced MS and BMF. TMZ induced MS was defined as a decrease in at least 2 cell lines below normal that was clinically attributable to TMZ and persisted for > 28 days after the last TMZ dose. Severe MS was defined as bone marrow cellularity of <25% or at least 2 of: a) reticulocyte count <40,000/µL b) absolute neutrophil count (ANC)<500/ µL c) platelets <20,000/µL. Very severe MS was defined by an ANC <200/ µL.

Results: A total of 15 Mayo patients and 10 published cases were identified. The baseline characteristics of patients are shown in table 1. The median interval between initiation of TMZ and diagnosis of MS was 48 days (range 26-121). The hematologic outcomes of the group are shown in table 2. Seventeen patients developed neutropenic fever, 4 had severe sepsis, and 6 experienced bleeding complications. Kaplan-Meier recovery curves for each cell line are shown in the figure. Myelosuppression was fatal in 3 cases. Most patients recovered their ANC (median-62 days, range 28-180). Persistent thrombocytopenia was seen in 19 patients, without evidence for clonal evolution or myelodysplasia. Four patients remain dependent on growth factor support (G-CSF and thrombopoietin agonists).

Conclusion: MS is usually a non-fatal complication of TMZ. Most cell lines recover spontaneously and adequately with the exception of platelets, which can remain significantly suppressed for prolonged periods. Infectious complications are very common. Most patients respond to supportive care, including the use of growth factors.

Figure 3 Figure 3.

Disclosures

No relevant conflicts of interest to declare.
Serum IL- 10 As a Predictive Marker of Response to Chemotherapy in Advanced Stage Hodgkin Lymphoma
Authors: Allahloubi Nasr Mohamed A. , Abdelhamid Thoraya Mohamed , Shamaa Sameh S. , Abd El Ghaffar Hassan , Saied Wageeh El-Sebaey , Awad Mohamed
Published: October 2019

Background: Cytokines and chemokines produced by HRS cells and the surrounding microenvironment are believed to be involved in the pathogenesis of HL by modulating immune functions and inflammatory responses.Among those cytokines; IL-10 seems to be most relevant for the shaping of a microenvironment favoring survival of HRS cells. Elevated IL-10 serum levels were even associated with a worse clinical outcome in patients with HL.

The aim of this study is to show the prognostic and predictive relevance of serum IL10 in advanced stage HL.

Patients and method: the study included 60 patients with advanced stage HL, treatment regimen was ABVD regimen (n=30) or BEACOPP baseline regimen (n=30). Serum IL10 assay employs the quantitative sandwich enzyme immunoassay technique. Evaluation of IL10 level was done at diagnosis then after 4 and 8 cycles of chemotherapy.

Results: The median agewas 41 years (range 20-65), 37 males (61.7%) and 23 females (38.3 %) were included. For patients treated with ABVD regimen, complete response (CR) was achieved in 18 cases (60%) compared to a CR rate of 66.7% for the BEACOPP regimen (CR= 20 cases) P=0.6. There was no significant difference between both treatment regimens regarding PFS and DFS (p=0.6 and 0.9 respectively). High pretreatment serum IL10 (>10pg/ml) showed significant relation to high LDH (p=0.001), male sex (p=0.018), more advanced stage IV versus IIB and III (p=0.0001) and the IPI score of 4 or more for advanced HL. This study showed that 83.3% of the patients with pretreatment serum IL 10 ≤ 10 pg/ml achieved CR compared to 45.2% of patients with IL-10>10 pg/ml did not achieved CR after 8th cycle of chemotherapy with (P= 0.035), Also the patients who achieved CR after 8 cycles had low level of IL-10 after treatment with a median of 9.35pg/ml (range 5.49-73.16) compared to a median of 30.66 (range 5.36-145.6) for patients who did not show CR (P<0.001). This significant relation between the CR rate and serum IL10 before and after treatment was also documented after the 4th cycles of chemotherapy (p=.006 and 0.003respectively). Our study found no significant relation between the IL-10 level prior to therapy and occurrence of relapse, disease progression or mortality.

Conclusion: There is a significant relations between high pretreatment serum level of IL-10 with poor response to chemotherapy after the 4th cycle and the 8th cycle of chemotherapy denoting that pretreatment serum level of IL- 10 is a good predictive marker of response.

Disclosures

No relevant conflicts of interest to declare.
Identification of a Subset of Peripheral T-Cell Lymphoma, Not Otherwise Specified, Characterized By FOXP3-Positive Regulatory T-Cell Phenotype, HTLV-1 Negativity and Poor Outcome
Authors: Pedersen Martin Bjerregaard , Hamilton-Dutoit Stephen Jacques , Bendix Knud , Nørgaard Peter H. , Michael Møller Boe , Raffeld Mark , Pittaluga Stefania , Steiniche Torben , Bergmann Anke , Vater Inga , Siebert Reiner , Chan Wing-Chung , d'Amore Francesco
Published: October 2019

Background: T-cell malignancies originating from regulatory T (Treg) cells are almost exclusively confined to human T-cell leukemia virus 1 (HTLV-1) associated adult T-cell leukemia/lymphoma (ATLL), although sporadic cases of other peripheral T-cell lymphomas (PTCLs) with hypothesized Treg derivation have been reported.

Patients and methods: We investigated a series of 169 well characterized PTCLs for the expression of Treg-cell phenotypic markers FOXP3, CD25 and CD4 by immunohistochemistry (IHC) using tissue microarray on formalin-fixed paraffin embedded tissue. Clinico-pathological data for patients enrolled were retrieved from the Danish Lymphoma Registry and medical records. Treg tumors were further invetsigated by Affymetrix OncoScan analysis and Illumina Infinium HumanMethylation450 BeadChips.

Results: Variable amounts of FOXP3-positive Treg cells were often (99% of the cases) found as part of the non-neoplastic cellular infiltrate. However, in five cases (3%) classified as PTCL, not otherwise specified (PTCL-NOS) the vast majority of what morphologically appeared to be the neoplastic cell population displayed a strong positivity for FOXP3, CD4, CD25 and CD3 suggesting a probable Treg origin of these cells. Cases were HTLV-1 negative and showed monoclonal rearrangements of the T-cell receptor genes. All cases were male older than 60 years and showed an aggressive clinical course with overall survival less than two years, despite all patients had low IPI-risk profile at the time of diagnosis. The PTCL with Treg phenotype showed a complex pattern of chromosomal imbalances. Moreover, as compared to normal T-cell subsets their DNA methylation profiles were mostly related to that of normal Treg cells but significantly differed from that.

Conclusions: We suggest that FOXP3-positve PTCL, in the absence of HTLV-1 infection, constitute a distinct entity separated from PTCL-NOS, occurring predominantly in elderly male patients and characterized by a highly aggressive clinical behavior. Further genomic analyses are ongoing. The identification and characterization of these cases may be useful to guide upfront therapeutic strategies.

Disclosures

No relevant conflicts of interest to declare.
CD180 Expression in B-Cell Lymphomas: A Multicenter Geil Study
Authors: Mayeur-Rousse Caroline , Guy Julien , Miguet Laurent , Bouyer Sabrina , Genevieve Franck , Robillard Nelly , Solly Francoise , Maar Aida , Bene Marie-C , Mauvieux Laurent
Published: October 2019

CD180 is a Toll-Like Receptor homolog strongly expressed on normal human B-cells and involved in innate immune responses. Previous proteomic analyses on microparticles derived from mature B-cell neoplasms allowed us to identify CD180 as a marker of marginal zone lymphomas (MZL)(Miguet, Leukemia, 2013). Using flow cytometry on blood samples we showed that this protein is lost or underexpressed at the plasma membrane for almost all B-cell lymphomas except MZL. In order to confirm its clinical relevance, we conducted a prospective multicenter flow cytometry study in 5 French University Hospital laboratories, on behalf of the GEIL. Blood or bone marrow samples from 236 patients were studied (20 normal controls ; 74 chronic lymphocytic leukemia (CLL); 21 mantle cell lymphoma (MCL); 42 lymphoplasmacytic lymphoma (LPL); 13 follicular lymphoma (FL) ; 58 MZL, 14 of which with numerous villous lymphocytes; 8 hairy cell leukemia (HCL)). Analyses were performed either on FACSCanto II (BD Biosciences, 3 centers) or on Navios (Beckman Coulter, 2 centers) instruments. Harmonization process was performed using Rainbow beads (Spherotech).

For the CLL group, CD180 Median fluorescence (MdFI) in each center was not significantly different (Anova test, p>0.05). Instruments’ harmonization was therefore effective enough to obtain similar data from all centres. In the whole cohort, CD180 was significantly less expressed in the group of lymphomas -including CLL, MCL, LPL and FL- than in controls (Mann-Whitney test, p<0.05). Conversely, in the group of MZL and HCL, CD180 MdFI was not different from those of controls (Mann-Whitney test, p>0.05) but significantly higher than in CLL, MCL, LPL and FL (Mann-Whitney test, p<0.0001). Distinction between MZL and lymphomas with numerous villous lymphocytes was possible (Mann-Whitney test, p=0.0012) but not between MZL and HCL. ROC curve analysis determined a CD180 MdFI threshold of 1800 which allow the positive diagnosis of MZL with a sensitivity of 77% and specificity of 92%.

These results underline the efficiency of CD180 as a single positive and robust marker for MZL diagnosis, and confirm that between centers and between instruments harmonization is largely feasible in routine practice as published recently (Solly F et al. Cytomery part A, 2013). It should be emphasized that among the group of lymphomas with intense expression of CD180, all interestingly originating from the spleen, those with numerous villous lymphocytes display the highest expression. We described for the first time in this study the strong positivity of CD180 in HCL. Anti-CD180 antibody may be included in diagnosis combination markers in order to improve the diagnosis of chronic B-cell malignancies

Disclosures

No relevant conflicts of interest to declare.
Exome Sequencing Identifies Somatic Mutations of DDX3X in Natural Killer/T-Cell Lymphoma
Authors: Jiang Lu , Gu Zhao-Hui , Yan Zi-Xun , Zhao Xia , Xie Yin-Yin , Zhang Zi-Guan , Pan Chun-Ming , Hu Yuan , Cai Chang-Ping , Dong Ying , Wang Li , Shen Yang , Meng Guoyu , Zhou Jian-Feng , Hu Jian-Da , Wang Jin-Fen , Yang Lin-Hua , Zhang Feng , Wang Jian-Min , Wang Zhao , Peng Zhi-Gang , Chen Fang-Yuan , Sun Zi-Min , Ding Hao , Huang Jin-Yan , Liu Yuan-Hua , Shi Jumei , Hou Jian , Yan Jin-Song , Shi Jing-Yi , Xu Lan , Li Yang , Lu Jing , Zheng Zhong , Xue Wen , Zhao Wei-Li , Chen Zhu , Chen Sai-Juan
Published: October 2019

Natural-killer/T cell lymphoma (NKTCL) is a malignant proliferation of CD56+/cytoCD3+ lymphocytes and constitutes a heterogeneous group of aggressive lymphoma prevalent in Asian and South American populations. NKTCL represents a distinct clinicopathologic entity of non-Hodgkin’s lymphoma, characterized by male predominance, strong association with Epstein-Barr virus (EBV) infection, prominent tissue necrosis and aggressive clinical course. However, molecular pathogenesis of NKTCL remains largely elusive.

Here we identified somatic mutations by whole-exome sequencing in 25 NKTCL patients and extended validation through targeted sequencing in an additional 80 cases. Functional experiments including RNA unwinding test, colony forming assay, cell proliferation assay and gene expression profiling were also performed.

Overall, 50.5% of NKTCL patients displayed somatic mutations of RNA helicase family, tumor suppressors (TP53 and MGA), and/or epigenetic modifiers (MLL2, ARID1A, EP300 and ASXL3). Recurrent mutations were most frequently discovered in RNA helicase gene DDX3X (21/105 cases, 20.0%). Mutations of DDX3X were seldom overlapped with those of TP53. Functionally, DDX3X mutants exhibited reduced RNA unwinding activity and enhanced cell proliferation. Similar stimulatory effect on cell proliferation was observed in cells transfected with specific siRNA targeting DDX3X. Gene expression profiling revealed an association of DDX3X mutations with activation of NF-kB and MAPK pathways. The clinical follow-up data showed that DDX3X-mutated patients presented a poor prognosis.

Our work suggests the heterogeneity of gene mutational spectrum of NKTCL and provides a potential therapeutic target for relevant cases.

Disclosures

No relevant conflicts of interest to declare.
Utility of F-18 FDG PET/CT for the Diagnosis and Therapeutic Management of Extranodal Natural Killer (NK)/T-Cell Lymphoma, Nasal Type
Authors: Xu Bing , Li Rongwei , Shi Pengcheng , Dong Huijuan , Guo Xutao , Fan Zhiping , Huang Fen
Published: October 2019

Background: Extranodal natural killer (NK)/T-cell lymphoma, nasal type (ENKTL) is quite rare in Western populations, but relatively common in East Asia (especially China) and Latin America. ENKTL is an aggressive disease with poor prognosis, requiring risk stratification. Similar symptoms, such as nasal obstruction and purulent nasal discharge, are found in patients with nasal NKTL and in patients with chronic rhinosinusitis. The images from contrast-enhanced computed tomography (CT) frequently show no prominent mass. As a result, the diagnosis is often delayed because it is frequently misdiagnosed as sinusitis. F-18 FDG PET/CT is a powerful imaging tool for diagnosis, staging, and evaluation of therapeutic effect in oncology.

Aims: This study was designed to investigate the value of FDG PET/CT in the therapeutic management of extranodal natural killer (NK)/T-cell lymphoma, nasal type.

Methods: A total of 26 patients with NK/T-cell lymphoma, nasal type were diagnosed according to morphologic and immunophenotypic criteria as specified

in the World Health Organization (WHO) classification. All patients underwent FDG PET/CT and clinical information was obtained by review of medical records.

Results: In 26 cases, all nasal/extranasal lesions were FDG-avid evidented to be neoplasm on CT scan and histopathological examinations. FDG-avid lesions in nasal/maxillary areas were uniformly more localized than demonstrated on CT scan, suggesting soft tissue masses on CT were partly due to inflammatory reaction. Among the 26 patients with definite diagnosis, 9 patients were re-staged on the basis of F18 FDG PET/CT with 5 patients down-staged and 4 patients up-staged. Statistical difference of the standardized uptake values (SUV) after 6 courses of chemotherapy and/or radiotherapy between the complete remission (CR) group and the partial remission (PR) group can be found (4.1±2.3 versus 7.8±1.7,P=0.006). The SUV value between pre-reatment and post-treatment were also of statistical significance in 22 patients (11.4±6.2 versus 5.6±2.1,P = 0.000).At a follow up of 18 months, patients got CR had a longer survival time than those got PR, stable disease (SD) or progress disease (PD) in 20 patients out of 26 patients.(median survival:310 days vs 284 days,(95% CI: 0.7156 to 1.365, p<0.05).

Conclusions: Our preliminary study suggests that FDG PET/CT can provide more accurate information on the diagnosis, staged and therapeutic response assessment in extranodal natural killer (NK)/T-cell lymphoma, nasal type. FDG PET/CT can be an invaluable imaging modality in this clinical setting. Further investigation with large patients enrollment is warranted.

Disclosures

No relevant conflicts of interest to declare.
Long-Term Remission (>3 years) after Rituximab Monotherapy Used for Patients with Relapsed Indolent Non-Hodgkin Lymphomas (NHL)
Authors: Michallet Anne-Sophie , Beckerich Florence , Bouafia-Sauvy Fadhela , Espinouse Daniel , Salles Gilles , Coiffier Bertrand
Published: October 2019

Background:

Rituximab has demonstrated significant clinical efficacy in the treatment of NHL, particularly in combination with chemotherapy, and its use has dramatically changed the treatment and outcome of both indolent and aggressive B-cell NHL over the past decade. Furthermore, consistent toxicity data have been obtained with a safe and tolerable profile in a large majority of patients.

Aim:

The objective of this retrospective study is to evaluate the long-term efficacy of rituximab monotherapy (4 weekly infusion at induction followed by 4 infusion every 2 months as consolidation) used for patients with relapsed indolent NHL.

Results:

From May 1998 through January 2011, Among 919 patients with indolent NHL treated in our department, 488 (53%) relapsed and were treated with rituximab alone (first and later relapse). 126 (26%) responded and were still in response 2 years later. These 126 patients (68 (54%) males and 58 (46%) females with a median age of 61 y; range: 17-94) are the subject of our analysis. 24% were over 70 years old, 45% were in first relapsed and 25% in later relapsed. Only 16% of the population have more than one co-morbidities (cardiac and or renal respectively); 79% a normal PS (0-1) with only 4 patients having a PS >2; 80% no bulky disease but 72% a disseminated disease (73% stage III and IV). In this study, 60 (48%) patients had a follicular lymphoma with 50% at an intermediate risk group according to the FLIPI score ; 20 (16%) a marginal zone lymphoma, and 43 (34%) a small lymphocytic or lymphoplasmacytic lymphoma and 3 patients other characteristics of lymphoproliferative disorders. After rituximab monotherapy, 55% of the entire cohort was in complete response (CR) and 31% in CRu or partial response and 5% in stable disease. Among these 126 patients, 74% progressed later than 3 years (38% later than 5 years). With a median follow-up of 6.5 years, 4-year and 8-year PFS were 70% and 30%, respectively. 18% had transformation into Richter syndrome and 12 patients have died.

Conclusion: Rituximab monotherapy is an effective therapy in selected relapsed indolent NHL and allows long-term response. This strategy could be used as “spare of chemotherapy” which is an important question today, especially in indolent not curable disease.

Disclosures

No relevant conflicts of interest to declare.