bluuebunny/crossref_metadata_2025 huggingface dataset:
PD-L1 Expression Is Low in Large B-Cell Lymphoma with MYC or Double-Hit Translocation
Authors: Elbæk Mette Vestergaard , Pedersen Mette Ø , Breinholt Marie Fredslund , Reddy Anupama , Love Cassandra , Clasen-Linde Erik , Knudsen Helle , Nielsen Signe Ledou , Gang Anne Ortved , Hogdall Estrid , Dave Sandeep , Norgaard Peter H.
Published: October 2019

Introduction: In large B-cell lymphoma (LBCL) MYC translocation and MYC/BCL2 or BCL6 double hit (DH) is associated with poor prognosis and there is an unmet need for novel treatment targets in this patient group. Treatment targeting the PD-L1/PD-1 pathway has been successfully introduced in Hodgkin lymphoma and solid cancers but are still poorly elucidated in LBCL. PD-L1 expression might predict response to treatment targeting the PD-L1/PD-1 pathway. We therefore investigated the relationship between PD-L1 protein and mRNA expression levels and MYC and DH translocation in LBCL.

Material and Methods: MYC, BCL2 and BCL6 translocations were detected by fluorescent in situ hybridization in tissue samples from 130 patients randomly selected from two cohorts of patients with LBCL: 49 patients with MYC translocation of whom 36 patients had DH and 81 without MYC translocation. PD-L1 protein expression was detected by immunohistochemistry (IHC) in tissue samples from 77 patients and PD-L1 mRNA expression by next-generation RNA sequencing (NGS) in another 77 patients. 24 patients overlapped, i.e. were analysed with both IHC and NGS. Nonparametric tests were performed to evaluate intergroup differences.

Results: PD-L1 protein expression level was lower in patients with MYC translocation (n=42, median=3,3%, IQR 0,0-10,8) or DH (n=31, median=3,3%, IQR 0,0-10,0) compared to patients with no MYC translocation (n=35, median=16,7%, IQR 3,3-30,0) or DH (n=46, 13,3%, IQR 2,5-30,0), P=0.004 and P<0,001 respectively (Fig.1). PD-L1 mRNA expression was also significantly lower in patients with MYC translocation or DH, P=0,001 and P=0,006 respectively. Higher PD-L1 protein and mRNA expression levels were associated with non-GC-type compared to GC-type DLBCL, P= 0,004 and P=0,002 respectively.

Conclusions: We report a highly significant association between low PD-L1 expression and MYC and DH translocation in patients with LBCL. Our findings may indicate that patients with MYC or DH translocation may benefit less from treatment with PD-L1/PD-1-inhibitors compared to patients without these translocations. This should be evaluated in larger, prospective, consecutive trials.

Disclosures

No relevant conflicts of interest to declare.
Regulation of Homeostatic and Malignant B Cell Development By the Tetraspanin CD53
Authors: Greenberg Zev J. , Monlish Darlene A. , Bartnett Rachel L. , Schuettpelz Laura G.
Published: October 2019

Acute lymphoblastic leukemia (ALL) is the most frequent pediatric malignancy, most commonly originating from the transformation of progenitor cells of the B cell lineage (B cell precursor-ALL; BCP-ALL). Treatment of patients with high-risk or relapsed disease is difficult and prognosis remains poor in pediatric patients, with an even worse survival rate for adult BCP-ALL. Previous studies have shown an association of enhanced CD53 expression with many B cell malignancies, suggesting upregulation of CD53 may be implicated in carcinogenesis or maintenance of malignant cells. CD53 is a member of the tetraspanin family of transmembrane proteins, classically involved in cell adhesion, proliferation, and survival, and expressed exclusively on hematopoietic cells. While several studies have implicated a role for CD53 in regulating mature B cell proliferation, its role in early B cell development is not yet known. To elucidate the contribution of CD53 to normal and malignant B cell development, we have generated a CD53 knockout mouse.

In our CD53-/- mouse, we observe no differences in total white blood cell counts, yet the fraction of peripheral blood B cells is significantly reduced by 31% compared to wild-type (WT) controls (28.3% vs. 19.5%; p<0.005). During homeostatic B lymphopoiesis, CD53 increases through development, beginning at the pre-pro-B cell stage and reaching highest expression on mature B cells. Further investigation into the loss of B cells revealed that immature pre-B cells in the bone marrow and mature B cells in the spleen and lymph nodes are significantly diminished upon loss of CD53, resulting from increased apoptosis in CD53-/- mice. B cell differentiation of CD53-/- hematopoietic stem cells (HSCs) in vitro corroborates the dependence on CD53 for normal differentiation, as CD53-/- cultures have 26% fewer B cells than controls (p=0.033). Investigation into the signaling differences between WT and CD53-/- B cell progenitors by mass cytometry (CyTOF) suggests that decreased PI3K/Akt and MAPK signaling could be driving this loss.

With the observed loss of both B cell progenitors and mature B cells in CD53-deficient mice, CD53-/- mice were recently crossed to Eμ-Myc transgenic mice, a model of B-lineage leukemia/lymphoma, to generate WT, CD53-/-, Eμ-Myc+;CD53+/+, and Eμ-Myc+;CD53-/- groups to assess whether loss of CD53 alters the pathology or survival of these mice. As observed in human patients, moribund Eμ-Myc+ mice significantly upregulate CD53 on malignant cells, suggesting a potential role for CD53 during pathogenesis. Ongoing experiments are aimed at elucidating the mechanism by which CD53 promotes homeostatic B cell development and determining the potential of CD53 as a therapeutic target for B lineage malignancies.

Disclosures

No relevant conflicts of interest to declare.
Functional Analysis of the FOXO3 Gene on the Induction of Fetal Hemoglobin in K562 Cells
Authors: Albuquerque Flávia Peixoto , Franca-Neto Pedro Luis , Franca Rafael de Oliveira , Araujo Aderson da Silva , Costa Fernando F. , Bezerra Marcos , Lucena-Araujo Antonio R.
Published: October 2019

The β-hemoglobinopathies are known as genetic disorders of high impact and wide distribution worldwide, with chronic and variable clinical prognosis. These pathologies have their symptoms and sequelae alleviated by increased levels of fetal hemoglobin (HbF; α2γ2). The Hydroxyurea, a drug used for the treatment of β-hemoglobinopathies, is effective for the maintenance and control of the development of the main clinical manifestations, however, it needs some prerequisites for its use, which indicates the need for new therapeutic approaches for this group of individuals. The family of Forkhead Box O transcription factors (FOXO), composed of FOXO1, FOXO3, FOXO4 and FOXO6, plays a key role in the regulation of several biological processes such as: regulation of the cell cycle, modulation oxidative stress, regulation mechanisms of response to DNA damage, controlling apoptosis and inflammatory response. Among members of the FOXO family, FOXO3 has been described as a physiological regulator of the erythroid maturation process and indicated as a positive regulator of HbF levels. In the present work, we evaluated the expression levels of the FOXO3 gene in patients with hemoglobinopathies and, in an attempt to better understand the pathophysiology of the disease, we performed a subcloning of the FOXO3 gene into a lentiviral expression system for its constitutive expression in vitro and subsequent evaluation of its activity as a possible transcription factor for γ globin genes (HBG1 and HBG2). In our clinical study, FOXO3 expression was significantly higher in patients with sickle cell anemia when compared to healthy donors (HbAA) (P <0.001) . In addition, a differential expression of FOXO3 was associated with the development of lower limb ulcer (P <0.005). Although we observed a tendency to increase FOXO3 expression in patients with 2 or more clinical manifestations, this association was not statistically significant (P = 0.06). The same was observed when we compared the number of clinical complications presented by patients last year with HbF levels: patients with 2 or more complications had lower levels of HbF, although this difference did not reach statistical significance (P = 0.182). There was no correlation between FOXO3 expression levels and HbF dosage (r = -0.12). Because of the activity of the FOXO3 gene in the control of oxidative stress, experiments were performed to evaluate the ROS and NO rates in the cell culture and a significant role was found, the FOXO3 increasing NO production (P = 0.0001) as well as in control of ROS produced in mitochondria.

Disclosures

No relevant conflicts of interest to declare.
Alternate Day Oral Iron Dosing in Iron Deficiency Anemia
Authors: Bulbul Ajaz , Araujo Mino Emilio , Dake Vyas , Bautista Adriana , Lentkowski Lisa , Khorsand-Sahbaie Masoud
Published: October 2019

Introduction

Practice guidelines for Iron deficiency anemia (IDA) suggest taking ferrous iron in divided doses. Recent studies suggest that split daily dosing may increase serum hepcidin which reduces iron bioavailability. Adherence to oral iron supplementation (OIS) can also be a barrier to treatment. In practice iron dosing varies significantly with unclear evidence of benefit from a particular dosing regimen.

Methods

This is a retrospective study evaluating outcomes of different schedules of OIS in 146 patients with IDA (Hb <12.2 and/or ferritin of < 30 ng/mL) treated between June 2017-June 2018. Patients with multifactorial anemia were excluded. Descriptive statistics and Chi-square were used for analysis.

Results

The mean age was 66.8 ± 1.3; women constituted 70% of the cohort (M/F 44/102). Mean Hb was 11.59 ± 0.12; Median ferritin was 22 ng/ml (IQR 10-63). Four different schedules of OIS evaluated were every other day (QOD) 60% (88/146), daily (QD)15% (22/146), twice daily (BID) 12%(18/146) and three times a day (TID) 12% (18/146). After one month of OIS a mean increase in Hb was (0.44 mg/dl + 0.04). GI toxicity occurred in 10.2% (15/146), therapy discontinuation in 4.8% (7/146) and IV iron was required in 9.6 % (14/146) of all cases. Among patients without GI toxicity 65% (85/131) were on QOD vs other schedules (X² 11.7 p=0.008), 63% (87/139) were compliant on QOD (X² 9.05 p=0.029). Salvage IV iron was not required in 64% (84/132) of QOD patients (X² 22.7 p=<0.001). One month post therapy, patients on QOD schedule had ≥1 g/dl improvement in 38% (10/26) (X² 9.18 p= 0.027) and increase of >0.5g/dl in 68% (69/102) of cases (X² 9.63 p= 0.022).

Conclusions

Alternate day iron dosing may optimize iron absorption and is possibly a better tolerated regimen. Larger prospective studies need to confirm these findings

Disclosures

No relevant conflicts of interest to declare.
Programmed Cell Death Receptor (PD-1), PD-1 Ligand (PD-L1) Expression in Philadelphia Chromosome Negative Myeloid Neoplasms
Authors: Chen Chi , Pandey Anita , Kundra Ajay , Kodali Sreenath , Wong Ching , Cheung Tony W. , Gotlieb Vladimir K. , Joseph Gardith , Tribie Sophia , Wang Jen-Chin
Published: October 2019

Introduction : PD-1 and PD-L1 pathway inhibitor therapy has been extensively explored in the field of oncology treatment recently, however there is no information so far about its usefulness in myeloid neoplasms (MPN). We have previously reported on PD-1 and PD-L1 expression in Philadelphia chromosome negative myeloid neoplasm (MPN), but now that we have accumulated more cases , we wish to report the final results.

Materials and Methods: 63 patients with Philadelphia chromosome negative MPN in total were studied which included 16 MF (14 PMF, 2 post-PV-MF, 2 post-ET-MF), 29 ET, 18 PV, and 25 normal volunteer controls.

Flow Cytometry Studies: Leukocytes were obtained from whole blood after RBC lysis using RBC lysis buffer (Qiagen). To quantify surface bound PD-1 and PDL-1, the leukocytes were incubated with CD279-APC and CD274-PE, as well as CD4-FITC, CD8-FITC, CD14-PCP, or CD34-FITC (BD Biosciences) and were subjected to flow cytometric analysis.

Results: PD-1 and PD-L1 positive cells were expressed as a percentage in gated CD4bright, CD8+, CD14+, or CD34+ cells. We found that 1) there was no difference in PD-1 or PD-L1 levels among different groups of MPN, but there was a significant difference when they were grouped of PV, ET and MF as MPN and compared with controls in all the immune cells including CD4+, CD8+, CD14 + and,CD34+ cells.(Representative of CD4+ shown inFig(1a). 2) High expression levels (more than 50% cell positivity) was found on the CD34+ cells with PD-1 and PD-L1 expression of 20% and 26% in MPN patients respectively ( Fig 1b,1c) . 3) We also found that there was no correlation between of the PD-1 or , PD-L1 expression levels with clinical features including such as WBC counts , platelet counts, and hemoglobin levels, , or presence or absence of the JAK2 , MPL, or CALR gene mutations or splenomegaly

Conclusion: We have shown elevated increased expression levels of PD1 and PDL-1 on the immune cells and high expression ( more than 50%) in the CD34+ cells , suggest that PD-1 or , PD-L1 pathway inhibitor therapy maybe worthwhile in Philadelphia chromosome negative Ph(-) MPN ..

Disclosures

Wang: celgene: Other: clinical trial.
The Zygosity of JAK2V617F Determines the Disease Entities of Myeloproliferative Neoplasms By Modulating Erythropoiesis but Not Megakaryopoiesis
Authors: Takei Hiraku , Edahiro Yoko , Li Lihua , Mizukami Yoshihisa , Imai Misa , Morishita Soji , Ohsaka Akimichi , Araki Marito , Komatsu Norio
Published: October 2019

Philadelphia-chromosome negative myeloproliferative neoplasms (MPNs) consists of different clinical entities that include polycythemia vera (PV) and essential thrombocythemia (ET). Despite of differential clinical features, JAK2V617F mutation is found in both PV and ET, leaving the cause of differential biological phenotypes by an oncogene obscure. Previously, studies have shown that higher allele frequencies or expression of JAK2V617F are associated with PV symptom/features in patients or model animals, respectively, suggesting that the copy number of JAK2V617F modulates hematopoietic cell differentiation and thus exhibits differential clinical features. However, this remained elusive in human hematopoiesis. To examine the impact of the zygosity of JAK2V617F allele on hematopoietic differentiation in human cells, we established induced pluripotent stem cells (iPSCs) harboring heterozygous- and homozygous-JAK2V617F mutation using genome-editing techniques from normal iPSCs. The introduction of JAK2V617F mutation with one or two copies did not alter the pluripotency of iPSCs and their capacity to differentiate into hematopoietic stem/progenitor cells (HSPCs) in vitro. When we induced hematopoietic cell differentiation from HSPCs, factor-independent erythropoiesis and megakaryopoiesis were induced in both heterozygous and homozygous JAK2V617F-HSPCs. Furthermore, homozygous JAK2V617F-HSPCs showed higher potential for erythropoiesis compared to the ones with heterozygous JAK2V617F, while the zygosity of JAK2V617F showed less effect on megakaryopoiesis. To further understand the molecular mechanism of hematopoietic cell differentiation modulated by differential copy number of JAK2V617F, we analyzed the activation of JAK-STAT signal by immunoblot analysis. The activation of JAK-STAT signals was more prominent in HSPCs harboring homozygous JAK2V617F, than those with heterozygous JAK2V617F. This suggested that the level of JAK2 phosphorylation was positively correlated with the copy number of JAK2V617F. These observations implied followings: 1) heterozygous JAK2V617F was sufficient to promote the development of MPN by inducing the factor-independent erythropoiesis and megakaryopoiesis; 2) the zygosity of JAK2V617F determined the disease phenotypes of MPNs by modulating erythropoiesis but not megakaryopoiesis; and 3) the homozygous JAK2V617F increased JAK-STAT signaling in HSPCs, promoting an increased erythropoiesis.

Disclosures

No relevant conflicts of interest to declare.
Extracellular ATP and CD39 Regulates Mitochondrial Function and Cytarabine Resistance through Intrinsic PKA-ATF-PGC1a Pathway in Acute Myeloid Leukemia
Authors: Sarry Jean-Emmanuel , Recher Christian , Aroua Nesrine
Published: October 2019

Relapses in acute myeloid leukemia (AML) are caused by chemoresistant leukemic populations and new therapeutic approaches that specifically target these cells are urgently needed. Based on transcriptomic analyses of relevant PDX preclinical model of the resistance to cytarabine (AraC) and of the residual disease in patients, we identified ecto-nucleoside triphosphate diphosphohydrolase-1 CD39 (ENTPD1) overexpressed in the residual leukemic cells in vivo after chemotherapy. By flow cytometry, we confirmed that AraC increased cell surface CD39 expression in AML cell lines in vitro and in vivo as well as in 24 diverse patient-derived xenograft models. We further observed this increase in 100 patients at 35-days post-intensive chemotherapy compared to their respective diagnosis. Interestingly, high CD39 expression on AML patients was associated with a worse response to AraC in vivo. Furthermore, we showed that FACS-sorted CD39high AML cells had increased mitochondrial mass and activity, and were resistant to AraC in vitro and in vivo. We demonstrated that CD39 downstream signaling pathway was dependent on cAMP-PKA-PGC1a-TFAM axis and its inhibition by H89 sensitized AML cells to AraC through the inhibition of mitochondrial OxPHOS biogenesis and function. Finally, pharmacological inhibition of CD39 ATP hydrolase activity or genetic invalidation of CD39 protein using two inhibitors or shRNA markedly enhanced AraC cytotoxicity in AML cell lines and primary patient samples in vitro and in vivo. Together, these results indicate CD39 as a new player of the intrinsic chemoresistance pathway and a new therapeutic target to specifically overcome AraC resistance and eradicate these leukemic cells responsible for relapses in AML.

Disclosures

No relevant conflicts of interest to declare.
Prognostic Value of Pre-Therapy Serum Uric Acid in Diffuse Large B-Cell Lymphoma
Authors: Qian Shenxian , Yiwei Li
Published: October 2019

Background: High levels of serum uric acid (SUA) have been shown to associate with negative clinical outcome in various malignancies. This study investigates whether SUA, at the time of diagnosis, has a prognostic significance in patients with diffuse large B-cell lymphoma (DLBCL).

Methods: We retrospectively evaluated 167 Chinese patients with newly diagnosed DLBCL under rituximab (R)-CHOP or CHOP-like immune-chemotherapy from January 2008 to July 2016. The optimal cutoff value of SUA was determined by applying receiver operating curve (ROC) analysis. The prognostic influence of SUA and other factors were studied by Kaplan-Meier curve as well as univariate and multivariate Cox proportional analysis. The influence of SUA on the predictive accuracy of IPI score was subsequently calculated using the Harrell's concordance index (c-index).

Results: ROC analysis showed the cutoff value of SUA with best sensitivity and specificity was 6.4mg dl-1. Increased SUA level shown by Kaplan-Meier curve had a shorter progression free and overall survival (PFS and OS, p<0.001, respectively). In multivariate analysis, an independent significant association between elevated SUA levels and poor clinical outcome for PFS (HR=3.851; 95%CI 1.816-8.167, p<0.001) and OS (HR=4.007; 95%CI 1.884-8.523, p<0.001) was identified. The estimated concordance index, using IPI stratification measures (0.777), improved to 0.837 when SUA was integrated in.

Conclusions: In the present study, we concluded that increased SUA level at diagnosis is an independent predictor for worse clinical outcome in DLBCL patients. Integrating SUA to the IPI score might improve the survival prediction and risk stratification.

Disclosures

No relevant conflicts of interest to declare.
Effect of Proinflammatory and Homeostatic Cytokines on the Functionality of Human Double-Negative Regulatory T Cells
Authors: Peuser Moritz , Haug Tabea , Aigner Michael , Mackensen Andreas , Voelkl Simon
Published: October 2019

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is often the only curative treatment option for patients with hematologic malignancies. Its therapeutic effect is mediated by donor T cells, recognizing and eliminating residual malignant cells of the patient. However, the main adverse effect of allo-HSCT is also mediated by these T cells: the so-called graft-versus-host disease (GvHD). A promising approach to treat GvHD is the use of immunosuppressive T cell populations to inhibit uncontrolled T cell activation. A novel regulatory T cell population described in the setting of allo-HSCT is the subset of TCRαβ+ CD4- CD8- double-negative (DN) T cells. In murine models infusion and/or activation of DN T cells specifically suppressed alloreactive T cells and prevented development of GvHD after allo-HSCT. Of interest, clinical studies in patients who underwent allo-HSCT revealed an inverse correlation between the frequency of circulating DN T cells and the severity of GvHD. We have recently demonstrated that human DN T cells like their murine counterparts strongly inhibit proliferation of alloreactive CD4+ and CD8+ T-cells. Here we asked whether proinflammatory and homeostatic cytokines associated with incidence and severity of GvHD after allo-HSCT affect the suppressive activity of DN T cells. We found that the proinflammatory cytokines IL-6, IL-18, TNF and IFN-g do not affect the suppressive activity of human DN T cells nor render alloreactive T cells insensitive to inhibition. In contrast, the homeostatic cytokine IL-15 significantly diminished the immune regulatory function of DN T cells. Further analyses demonstrated that both conventional CD4+ T cells and DN T cells express the functional IL-15 receptor which elicits downstream receptor signaling. Of importance, IL-15 highly induced Akt/mTOR signaling cascade in DN T cells, a pathway reported to reverse suppressive activity of regulatory cells. Together, our findings indicate that the homeostatic cytokine IL-15 but not proinflammatory mediators impair DN T cell-mediated suppression of allogeneic T cell responses. Further understanding of the mechanisms involved in human DN T-cell suppression may have important implications for using them as a cellular-based therapy to limit alloreactive immune Responses.

Disclosures

No relevant conflicts of interest to declare.
Severely Impaired Platelet Function Due to a Novel Mutation of P2Y12 from a Human Subject with No History of Bleeding
Authors: Kostyak John C , Dangelmaier Carol A , Mauri Benjamin R , Patel Akruti , Kunapuli Satya P.
Published: October 2019

Platelets mediate hemostasis through amplifying an initial stimulus and aggregating at a site of injury. The identification of bleeding phenotypes in humans has the potential to reveal new insight into the mechanisms behind platelet signaling, as well as pinpoint new potential therapeutic targets. In fact, the study of bleeding disorders in human subjects has led to the identification of a number of disease states and proteins important to platelet functional responsiveness.

We have recently identified a human subject (PDS25) with a platelet functional disorder that has a normal CBC, and no history of bleeding diathesis. However, platelets from PDS25 have virtually no response to even high concentrations of 2-MeSADP, even though P2Y12 protein expression appears unaltered (Figure 1). Further, platelet reactivity to high doses of agonist for other receptors, such as PAR-4 and GPVI, is normal. Low concentrations of these agonists, which typically rely on feedback for platelet reactivity, elicit an inhibited response using platelets from PDS25 compared to platelets from a healthy control. Consistently, Rap1b activity (Figure 2) was reduced in platelets from PDS25, while VASP phosphorylation was enhanced, suggesting that signaling from the P2Y12 receptor was interrupted by the heterozygous mutation. To determine whether it is the receptor or a downstream signaling component that is dysfunctional and because shape change in response to 2-MeSADP is normal in platelets from PDS25, we co-stimulated 2-MeSADP treated platelets with epinephrine to initiate signaling through Gz, which is very similar to Gi. We found that the response of platelets from PDS25 to 2-MeSADP can be rescued with the addition of epinephrine, suggesting that the signaling components downstream of Gi/zare intact. These data suggest that the aberrant reactivity observed in platelets from PDS25 is due to the P2Y12 receptor. Therefore, we performed a genetic analysis of P2Y12 from PDS25, which revealed a heterozygous mutation of D121N within the conserved DRY motif (Figure 3). We are currently evaluating how this heterozygous mutation confers such a strong phenotype.

Disclosures

No relevant conflicts of interest to declare.
Long-Term Risk Factors for Occult Cancer Detection in Patients with Unprovoked Venous Thromboembolism: A Post-Hoc Analysis of the Reverse Study
Authors: Cantor Nathan , Carrier Marc , Rodger Marc A. , Veillet-Lemay Gabrielle , Sabri Elham , Kovacs Michael J.
Published: October 2019

Background: The diagnosis of an unprovoked (i.e., in the absence of any risk factors) venous thromboembolism (VTE) may be a sign of an underlying cancer. Previous estimates of the incidence rate of occult cancer in patients with unprovoked VTE are as high as 5-10%. However, screening methods to detect cancer early in these patients are of limited clinical value and previous identification of risk factors focused on short-term cancer diagnosis. Therefore, this study will assess the long-term incidence and risk factors of cancer in unprovoked VTE patients.

Methods: This retrospective study is a post-hoc analysis of the REVERSE (Recurrent Venous Thromboembolism Risk Stratification Evaluation) study. Patient data was collected following an initial six-month course of anticoagulation therapy and were followed for a mean time period of 5.0 years (range 1 month-8 years). We performed univariable analysis to test the strength of association between each potential predictor variable and occult cancer diagnosis. Furthermore, using forward model selection we created a multivariable model with optimal predictive value for cancer diagnosis.

Results: Among 663 patients presenting with unprovoked VTE, 38 (5.7%) subsequently developed a new cancer diagnosis during the follow-up period. The most common types of cancers diagnosed were prostate (21%) and colorectal cancer (16%). Univariate analysis revealed age > 65 (OR 2.59; 95% CI 1.31-4.49, P= 0.0036), elevated D-Dimer (OR 2.71; 95% CI 1.38-5.31, P=0.026) and pulmonary vein obstruction (PVO) score (OR 4.34 95% CI 1.26-14.92, P=0.023) as the strongest predictors of occult cancer. Only D-Dimer (Adj.-OR 2.72 95% CI 1.39-5.32) remained in the multivariate model and no other factor significantly improved the model's fit.

Conclusion: Patients with an unprovoked VTE remain at an elevated risk of developing cancer following anticoagulation therapy. Age > 65 years old, elevated D-Dimer, and PVO in pulmonary embolism patients are among potential risk factors for developing occult cancer. More studies are needed to validate these potentially important risk factors.

Disclosures

Carrier: BMS: Honoraria, Research Funding; Leo Pharma: Research Funding; Pfizer: Honoraria; Bayer: Honoraria. Rodger:Biomerieux: Research Funding. Kovacs:Daiichi Sankyo Pharma Development: Research Funding; Bayer: Research Funding.
Understanding the Role of Hyperdiploidy in Burkitt Lymphoma of Childhood: Biological and Clinical Correlates
Authors: Avgerinou Georgia , Papadhimitriou Stefanos , Stefanaki Kalliopi , Kattamis Antonis , Katsibardi Katerina , Kostopoulos Vassilios , Pavlidis Dimitrios , Polychronopoulou Sophia
Published: October 2019

Background: Burkitt lymphoma (BL) is the most common high-grade lymphoma in childhood. BL is typically associated with rearrangement of the MYC gene, on 8q24 chromosome region, and often accompanied by other aberrations, some of which are known to adversely influence the clinical behavior and prognosis of the disease. However, hyperdiploidy, though frequently detected cytogenetically, has not been convincingly linked to specific biological and clinical features of BL. We present here our experience with 8 pediatric hyperdiploid BL cases detected with molecular cytogenetics in the course of routine genetic investigation.

Materials and methods: The study included 44 children (35 boys, 9 girls), aged 1 to 15 years (median 6), diagnosed with histologically documented BL. In all cases, interphase FISH was performed on infiltrated bone marrow smears or touch imprints from samples of involved sites. FISH probes employed included sets of the detection MYC, BCL2, BCL6, IGH, IGK, and IGL rearrangement, -17/del(17p13), -9/(del9p21) and del(13q14)/del(13q34). Cases with over-representation of any of the chromosome regions targeted were further tested with the use of the appropriate centromeric/chromosome enumeration probe. Thus, this line of testing allowed for the detection of numerical aberrations of chromosomes 2, 3, 8, 9, 13, 14, 17, 18 and 22. Over-representation of at least three chromosomes, without evidence for any monosomy, was considered a hyperdiploidy criterion.

Results: MYC was found rearranged in all cases, in 38 together with IGH and in 6 of them with IGK or IGL rearrangment. 17p-, 13q- and 9p- were detected in 3, 4 and 2 cases, respectively. There were no cases with BCL2 or BCL6 rearrangements. With a median follow-up of 5.5 years, 4 relapses/deaths were observed. Hyperdiploidy was detected in 8 patients (18.2%), with involvement of at least 7 of the 9 chromosomes tested, at the level of trisomy to hexasomy. Hyperdiploid and non-hyperdiploid cases were comparable with regard to sex and age. However, all cases with IGK/IGL involvement, 17p-, 13q- or 9p-, and relapse/death were seen exclusively in the non-hyperdiploid group.

Conclusions: From the observations on this small group of patients, it seems that hyperdiploidy is rather common in childhood BL and, perhaps, represents a distinct clonal evolution pathway of the standard t(8;14)+ disease. It appears to be associated with a favorable prognosis. Further study on a larger number of cases is required to fully elucidate the clinical significance of the hyperdiploidy and whether it may be used as a marker of "low-risk" disease, by analogy to acute lymphoblastic leukemia.

Disclosures

Kattamis: Novartis: Consultancy, Honoraria; Vifor Pharma: Consultancy; CELGENE: Consultancy, Honoraria; ApoPharma: Honoraria.
The NCCN-IPI Score Is Prognostic of Survival in Patients with Peripheral T-Cell Lymphoma, Not Otherwise Specified
Authors: Beltrán Brady E. , Castro Denisse A. , Huerta- Collado Yesenia M. , Sotomayor Eduardo , Castillo Jorge J.
Published: October 2019

Introduction: Peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) accounts for 15-20% of NHL and is characterized by a poor survival. The IPI and PIT scores are prognostic factors in survival in B-cell and T-cell lymphomas, but not without limitations. The aim of this study is to evaluate the prognostic value of the NCCN-IPI score in patients with PTCL-NOS.

Methods: We included patients with a pathological diagnosis of PTCL-NOS who were diagnosed and treated at our institution between 1997 and 2017. IRB approval was obtained prior to research. Pathological samples were reviewed by hematopathologists at our institution to confirm the diagnosis. Pertinent clinicopathological data were collected through chart review and are presented using descriptive statistics. Survival curves were estimated using the Kaplan-Meier method and compared using the log-rank test. Univariate and multivariate Cox proportional-hazard regression models were fitted to evaluate hazard ratios (HR) for overall survival (OS).

Results: A total of 173 patients with diagnosis of PTCL-NOS were included in this analysis. The median age at diagnosis was 58 years (range 18-91 years) with a male predominance (58%). Clinically, 47% of patients were 60 or older, 52% presented with ECOG >1, 82% had elevated serum LDH, 74% had extranodal disease, and 34% had stage I/II and 66% had stage III/IV. IPI score distribution was low-risk in 55% of patients, low-intermediate in 36%, high-intermediate in 48% and high-risk in 33%. PIT score distribution was low-risk in 53%, low-intermediate in 67%, high-intermediate in 38% and high-risk in 14%. NCCN-IPI score distribution was low risk in 40%, low-intermediate in 56%, high- intermediate in 63% and high risk in 11%. 22% of patients received CHOEP, 23% received CHOP, and 55% received other regimens. The overall response rate was 59%; 47% had a complete response and 12% had a partial response. The 5-year overall survival (OS) rate was 32% with a median OS of 12 months. PTCL-NOS patients with low, low-intermediate, high-intermediate and high-risk NCCN-IPI had 5-year OS rates of 46%, 36%, 26% and 0%, respectively (p<0.001). When compared with patients with low-risk NCCN-IPI, patients with low-intermediate (HR 3.2, 95% CI,1.1-9.5; p=0.044) and high-intermediate /high risk NCCN-IPI (HR 6.0, 95% CI 2.1-17; p=0.001) had worse OS.

Conclusions: We have validated the NCCN-IPI score as a prognostic tool in patients with PTCL-NOS. This work can serve to address future prospective designs that allow selection of groups of patients at greater risk and thus lead to more individualized therapy.

Figure. Figure.

Disclosures

Castillo: Janssen: Consultancy, Research Funding; Genentech: Consultancy; Abbvie: Consultancy, Research Funding; Pharmacyclics: Consultancy, Research Funding; Millennium: Research Funding; Beigene: Consultancy, Research Funding.
Comprehensive Geriatric Assessment in Elderly Patients with Aggressive Non-Hodgkin Lymphoma in Peru
Authors: Beltrán Brady E. , Motta Rodrigo , Gamarra María P. , Araujo Roger , Castillo Jorge J.
Published: October 2019

Introduction: The incidence of Non-Hodgkin Lymphoma (NHL) is increasing, especially in people over 60 years of age. This population usually has a worse prognosis, probably due to comorbidities, functional deterioration and decreased tolerance to treatment. Therefore, a pre-therapeutic evaluation would be important to make decisions. The Comprehensive Geriatric Assessment (CGA) is the recommended tool for this evaluation, but it is a complex process that demands time and resources. We performed a study to evaluate the characteristics of the CGA in patients >64 years of age with NHL and to determine which domains can constitute a simplified model.

Methods: This is a cross-sectional study with retrospective data collection of geriatric evaluations performed in >64 years with aggressive NHL (90% DLBCL, 10% PTCL) admitted to our institution between September 2015 and August 2017. Number of drugs, prescriptions in older adults was evaluated (STOPP), Lawton scale, Barthel scale, KATZ index, walking speed, Up and Go time (TUG), Mini-mental test, Yesavage scale, Gijón scale, Mini-nutritional, Geriatric syndromes (Incontinence, Falls, Pressure ulcers, Immobility, Sensory Deficits, Osteoporosis), and Accumulated Disease Scale in Geriatrics (CIRS-G). The CGA included 9 domains, and fragility was defined as deterioration in >2 domains. The different evaluations were compared with fragility. Multivariate models were constructed using logistic regression.

Results: We included 253 patients with an average age of 76 years. 62% had >1 affected domain, and 40% were considered fragile (>2 affected domains). In the bivariate analysis, age >85 years, and all the geriatric scales except STOPP, were strongly associated with fragility. The final model had 6 variables: Use of >5 drugs ( OR 773.1, 95% CI,4.5-132134.4;p=0.011), Lawton scale <7 (OR 385.2,95%CI, 20.1-7363.5;p<0.001), TUG >20 (OR 124.6,95%CI, 6.2-2489.8;p=0.002), Mini-mental <23 (OR 53.5,95%CI, 3.7-778.8;p=0.004),Yesavage scale >5 (OR 118.7,95%CI, 11.2-1254.5;p<0.001), and presence of at least one geriatric syndrome (OR 235.6,95%CI, 13.3-4158.7;p<0.001). Removing the Mini-mental minimally affected the model, but suppressing two or more variables does weaken the model.

Conclusions: In our cohort of patients older than 64 years with aggressive NHL, a model based on five measurements (i.e. number of drugs, TUG, Lawton scale, Yesavage scale and Geriatric Syndromes) could constitute an evaluation with an efficiency similar to a complete CGA.

Disclosures

Castillo: Pharmacyclics: Consultancy, Research Funding; Genentech: Consultancy; Millennium: Research Funding; Abbvie: Consultancy, Research Funding; Janssen: Consultancy, Research Funding; Beigene: Consultancy, Research Funding.
Dysfunctional CLEC-2 and GPVI-Mediated Signaling in Syk Y342F Knock-in Mouse Platelets
Authors: Kostyak John C , Mauri Benjamin R , Dangelmaier Carol A , Patel Akruti , Wright Monica , Reddy Haritha G , Eble Johannes A , Kunapuli Satya P.
Published: October 2019

Platelet activation is essential for hemostasis. Central to platelet activation are the signals transmitted through surface receptors like GPVI, the protease activated receptors (PARs), and C-type lectin-like receptor 2 (CLEC-2). GPVI is an ITAM-bearing receptor, while Clec-2 is a HemITAM-bearing receptor. Both ITAM and hemITAM-bearing receptors signal through spleen tyrosine kinase (Syk). Syk activity is dependent on phosphorylation of several residues including Y342, Y346, and Y519/520 (Y348, Y352, and Y525/526 in human). However, the importance of each residue on either phosphorylation of other residues or activation of Syk is unknown. Therefore, we produced a Syk Y342F knock-in mouse line via CRISPR/Cas9 to investigate the importance of Y342 on Syk-mediated signaling using a platelet system. Syk Y342F mice are viable, have no gross abnormalities, and their blood cell counts are normal. We were unable to detect any Syk Y342 phosphorylation following stimulation of Y342F platelets with the GPVI agonist collagen-related peptide (CRP) (Figure 1). Further, Platelet reactivity is reduced in response to the GPVI agonists CRP (Figure 2) and collagen, as well as the CLEC-2 agonist rhodocytin. When using CLEC-2 antibody as an agonist no response is observed in Syk Y342F mouse platelets even though WT littermate control mouse platelets responded well (Figure 3). Signaling initiated by either GPVI or CLEC-2 is also inhibited, as Syk Y519/520 phosphorylation is impaired. Furthermore, PLCg2 and SLP-76 phosphorylation are reduced in Syk Y342F platelets compared to platelets from littermate control mice following stimulation of either GPVI or CLEC-2. Although reactivity of Y342F platelets is clearly reduced compared to WT platelets, there was no difference in occlusion time following ferric chloride injury of the carotid artery. Similarly, there was no difference in mortality rate following pulmonary thromboembolism using Syk Y342F mice compared to WT littermate control mice. Nevertheless, these data demonstrate that phosphorylation of Y342 on Syk following stimulation of either ITAM or hemITAM-bearing receptors is important for Syk activation.

Disclosures

No relevant conflicts of interest to declare.
Impact of T-Cell Dose on Gvhd Risk after Allogeneic HLA-Matched PBSC Transplantation
Authors: Saad Ayman , Wang Tao , Hemmer Michael T. , Spellman Stephen R. , Arora Mukta , Lamb Lawrence S. , Hashmi Shahrukh K.
Published: October 2019

Background: Conflicting data exists on whether T-cell dose of allogeneic stem cell product influences transplant outcome.

Methods: Using CIBMTR database, we identified 2,736 adult patients who underwent first allogeneic HCT for AML/ALL/MDS (between 2008-2014) with PBSC from an HLA-identical sibling donor (MSD) or 8/8-matched unrelated donor (MUD). This cohort excluded ex-vivo and in-vivo T-cell depletion transplants. Correlative analysis was done between CD3 cell dose and risk of GVHD, relapse, NRM, DFS and OS.

Results: T-cell dose cutoff values were identified (using maximum likelihood estimation method) to identify differential risk of aGVHD (grade II-IV) in both MSD and MUD groups. A CD3 cell dose cutoff of 14 x107 cells/kg identified MSD/low CD3 (n=223) and MSD/high CD3 (n=1214) and a dose of 15 x107 cells/kg identified MUD/low CD3 (n=197) and MUD/high CD3 (n=1102). CD3 and CD34 cell doses did not correlate. Median CD3 cell dose were 11 and 29 in the MSD/Low and MSD/High groups, respectively, and 10 and 28 in the MUD/Low and MUD/High groups, respectively. In MSD/high CD3, cumulative incidence (CI) of D100-aGVHD (GII-IV) was higher (33% vs 25%) (P value =0.009) without influence on engraftment, severe aGVHD (GIII-IV), cGVHD, NRM, relapse, DFS, or OS. In MUD/high CD3, CI of D100-aGVHD (GII-IV) was higher (50% vs 40%) (p value=0.009) and so was the CI of 6-month-cGVHD (31% vs23%) (p value = 0.02) with no influence on engraftment, severe aGVHD, 2-year-GVHD, NRM, relapse, DFS, or OS. On multivariate analysis, both MSD (table 1) and MUD (table 2) groups failed to show a correlation between CD3 cell dose and aGVHD (GII-IV) (p value =0.1 and 0.07), cGVHD (p value=0.8 and 0.3) for both groups respectviely. Sub-analysis of CD4, CD8 and CD4/CD8 ratio failed to identify cutoff values predictive of transplant outcome. Using log-rank test, the sample size was, however, suboptimal to identify difference (with 80% power at 0.05% significant level) at these cutoff cell dose.

Conclusion: In this relatively small registry study, the CD3 cell dose in the PBSCT product did not influence the risk of acute or chronic GVHD or other transplant outcome when using matched sibling or 8/8 unrelated donors.

Disclosures

Saad: Actinium: Consultancy. Lamb:Incysus Therapeutics Inc.: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding.
A Prospective Phase II Study of Thalidomide, Cyclophosphamide and Prednisone in Newly Diagnosed Idiopathic Multicentric Castleman's Disease
Authors: Zhang Lu , Zhao Ai-lin , Feng Jun , Cao Xinxin , Zhang Yan , Zhu Tienan , Duan Minghui , Zhou Daobin , Li Jian
Published: October 2019

【Introduction】Idiopathic Multicentric Castleman's disease (iMCD) is a rare lymphoproliferative disorder without 'standard' treatment. In current study, we aimed to investigate the efficacy and safety of TCP regimen (thalidomide, cyclophosphamide and prednisone) in newly diagnosed iMCD patients.

【Methods】From Jun, 2015 to Jun, 2018, thirty-six newly diagnosed iMCD patients were enrolled. TCP regimen (thalidomide 100mg qn for 1 year and maintained with 100mg qn for the second year; cyclophosphamide 300mg/m2 weekly on d1, 8, 15, 22 out of a 28-day cycle; prednisone 1mg/kg twice a week on d1-2, d8-9, d15-16, d22-23 out of a 28-day cycle) was administrated until death or 'treatment failure'. Data from twenty-five patients whose were followed for at least 24 weeks were extracted and analyzed.

【Results】Among these 25 patients, the median age was 40 years (range 20-63 years), with a male : female ratio of 2.1:1. Two patients fulfilled the diagnostic criteria of TAFRO syndrome. After TCP treatment, 48.0% (n=12) patients achieved 'durable tumor and symptomatic response' for at least 24 weeks; 40.0% (n=10) patients were evaluated as 'treatment failure'; 12.0% (n=3) patients were considered as stable disease. The median symptom score was reduced from 14 (range 6-41) to 3 (range 0-20); the median Interleukin-6 level decreased from 21.3 pg/ml (range 7.4-865) to 9.15pg/ml (range 2.0-36.2). As for safety issues, one patient died from pulmonary infection and one patient suffered from Grade 3 rash. No other patients suffered from Grade 3 or above adverse events. With a median follow-up of 14 months (range 6-36), 8.0% (n=2) patients died: one patient died from progression of disease (severe anemia) and the other died from pulmonary infection after TCP administration. Median overall survival (OS) and progression-free survival (PFS) were not reached, with the estimated 3-year PFS and OS of 60.0% and 88.0%, respectively (Figure 1).

【Conclusions】TCP regimen, an inexpensive oral therapy, is an effective and safe first-line treatment option for newly diagnosed iMCD patients.

Figure 1. Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Increased Longevity of Continuous Bone Marrow Cultures and Radioresistance of Bone Marrow Stromal Cells from SOD193A ALS (Amyotrophic Lateral Sclerosis) Mice
Authors: Henderson Andrew , Fisher Renee , Epperly Michael , Shields Donna , Rigatti Lora , Donnelly Christopher , Watkins Simon , Greenberger Joel S.
Published: October 2019

Introduction: The SOD1G93A mouse model of ALS, demonstrates hind limb paralysis beginning at 90 - 100 days of age with stage 4 paralysis at 125 days of age and progressive neuromuscular loss.

Materials & Methods: To determine whether deficiency of functional SOD1 influenced parameters of hematopoiesis, long-term bone marrow cultures were established from ALS and control mice. Bone marrow stromal cell lines derived from LTBMCs were tested for clonogenic radiation survival. We tested the effect of bone marrow transplant after total body irradiation on delay of paralysis.

Results: SOD1G93A marrow cultures demonstrated significant increase in production of hematopoietic progenitor cells (p < 0.0001) and overall longevity of production of hematopoietic cells (p = 0.0354), and bone marrow stromal cell lines were significantly radioresistant (D0 = 1.33 ± 0.09, and ñ = 8.57 ± 1.8) compared to control C57BL/6J mice (D0 = 1.59 ± 0.11, p = 0.117; and ñ = 3.4 ± 0.4, p= 0.0466).

Total body irradiation and bone marrow transplantation with GFP+ donor marrow demonstrated a significant increase in paralysis free interval from 129.2 ± 3.0 to 240.7 ± 21.1 days (p = 0.0010), normalization of blood/brain barrier permeability, and increase in M2 marrow origin microglial cells in proximity to degenerating anterior horn cell/motor neurons. Isolated brain and spinal cord irradiation did not prolong the paralysis free interval (129.0 ± 2.7 days, p = 0.7748).

Conclusions: The results showing increased longevity of hematopoiesis in LTBMCs of marrow from mice displaying an absence of SOD1 and the radioresistance of derived bone marrow stromal cell lines represent two unexpected pleiotrophic effects of the SOD1 G93A genotype. Further studies will be required to determine how marrow transplant after TBI prolonged the paralysis free interval in these ALS mice.

Disclosures

No relevant conflicts of interest to declare.