bluuebunny/crossref_metadata_2025 huggingface dataset:
Interrogation of the Coagulation Cascade Acute Coronary Syndrome Using Novel Elisa-Based Assays for Single Protease Quantification
Authors: Posma Jens , van Oerle Rene , Fens Diane , govers Riemslag Jose , Mueller Julia , Heitmeier Stefan , ten Cate Hugo , Spronk Henri M.H.
Published: October 2019

Background: Acute coronary syndrome (ACS), the main contributor to myocardial infarction, is a thrombotic complication of atherosclerosis. Growing evidence supports a role for the intrinsic coagulation cascade in various thrombotic diseases. To gain more inside in the contribution of the intrinsic coagulation cascade in an ACS, we developed novel, ELISA-based assays for the quantification of Kallikrein, activated factor XII (FXIIa), FXIa, FXa, FIXa, and thrombin in complex with a physiological inhibitor.

Methods: During this prospective cohort study, blood from patients with a first ACS (n=61) was collected upon admission (before heparin injection(t=0), and after 1(t=1) and 6 (t=2) months. Plasma levels of Kallikrein-C1inhibitor (K-C1inh), FXIIa-C1inh, FXIa-C1inh, FXIa-α1 anti trypsin, FXIa-antithrombin (AT), FIXa-AT, FXa-AT, and thrombin-AT(TAT) were measured using newly developed ELISA's and compared to apparently healthy controls (n=60). Methods were validated according to the EP5-protocol for within- and -between run variability.

Results: The inter coefficients of variability for K-C1inh is 11.9%, FXIa-α1-AT 14.4%, FXIa-AT 13.7%, FIXa-AT 4.9%, FXa-AT 2.6% and TAT 8.6 %. Levels of FXIa-α1-AT, FXI- ΑΤ, FIXa-AT and TAT were all significantly elevated in plasma from ACS patients at time of the acute event compared to healthy control: FXIa-α1 anti trypsin respectively 202.8 (IQR 156.6 - 283.4) vs 29.0 pM (IQR 0.00 - 101.0); FXIa-AT levels were respectively 35.9 pM (IQR 23.9 - 43.4) vs 28.0 pM (IQR 23.3 - 34.0); FIXa-AT respectively 102.0pM (IQR 87.4 - 123.0) vs 90.5 pM (IQR 83.2 - 104.3); TAT respectively 5.59 (IQR 3.91 - 9.85) vs 2.26 pM (IQR 1.78 - 2.71)(Fig1). Plasma levels of FXIa-α1-AT, FXIa-AT FIXa-AT, and TAT decreased during follow up. Additionally, FXIa-α1-AT, FXIa-AT FIXa-AT, and TAT were higher in patients with ST-elevated myocardial infarction (STEMI) compared to subjects with non-STEMI or angina pectoris.

Conclusion: Elisa based profiling of the coagulation proteases provide a highly sensitive and reproducible method used to distinguish the different activation states of single proteases in the coagulation cascade. With these novel assays we show that FXIa-α1-AT, FXIa-AT FIXa-AT, and TAT are significantly elevated in patients with ACS. Additionally, patients with STEMI have a more pronounced hypercoagulable state than non-STEMI and angina pectoris patients.

Figure 1. Figure 1.

Disclosures

No relevant conflicts of interest to declare.
Prevalence of Hepatitis B, Hepatitis C and HIV in Sickle Cell Disease Patients from Oman
Authors: Alkindi Salam , AL-Umairi Nada , Pathare Anil
Published: October 2019

Background and Purpose: Oman is a country with an intermediate prevalence of Hepatitis B (HBV) carriers (2.8-7.1%, 20101), while the prevalence of HBV infection was 5.8% (20142). Hepatitis C (HCV) prevalence reported from Oman was 0.41% (20163). Further, a total of 2394 Human Immunodeficiency (HIV) infections were identified among Omanis (20134). Unfortunately, there are no studies on the prevalence of HIV, HBV and HCV in sickle cell disease (SCD) patients from Oman.

Objectives: This study was therefore performed to estimate the prevalence of HBV, HCV and HIV among SCD patients from Oman.

Methods: In a retrospective cross-sectional cohort study, medical records of all patients with SCD and sickle cell trait (SCT) patients who received blood transfusions between 2006 to 2017 were retrieved from the hospital information system. Specifically, data on HIV, HBV and HCV exposure were recorded to study the prevalence, following approval for the study from the hospital medical research and ethics committee.

Results: Amongst a total of 1000 patient (491 males and 509 females), 126 (12.6%) had anti-HCV antibodies (anti-HCV) based on serology and amongst these 52(5.2%) were PCR positive. Genotype 1 was the most predominant (n=36), followed by genotype 3& 4(n=7 each) followed by genotype 2(n=2). 23 (2.3%) SCD patients showed positive serology for hepatitis B surface antigen (HbsAg) and amongst these 16 (1.6%) were PCR positive. None of the SCD patients showed positive serology for HIV. Liver Ultrasound was normal in 788 patients (78.8%), whereas, 208(20.8%) had hepatomegaly and 4(0.4%)had liver cirrhosis. Further, only 2 patients amongst the 36 patients who had died had a hepatic cause.

Conclusion: This study provides comprehensive data on the prevalence of HBV and HCV infection among SCD patients and raises concerns regarding the HCV prevalence in these patients with its impact on chronic liver disease. Reassuringly, no case with HIV was observed.

Disclosures

No relevant conflicts of interest to declare.
Suppression of CUL4B Induces G1 Cell Cycle Arrest through the Modulation of AKT Signaling Pathway
Authors: Li Ying , Zhang Ya , Zhou Xiangxiang , Wang Xin
Published: October 2019

Introduction

T-cell lymphomas (TCL) are a heterogeneous group and one of the more aggressive of non-Hodgkin lymphomas. Previous studies have illustrated the poor prognosis in major propotion of patients due to refractory and recurrent TCL. CUL4B, a member of the CUL4B-RING E3 ubiquitin ligase complex, has been demonstrated to be involved in tumorigenesis and correlated to poor survival, contributing to tumor progression via several mechanisms, including epigenetic silencing of tumor suppressors. Although CUL4B was recently identified as a novel oncogene in several solid malignancies, the potential functional role of CUL4B in TCL are poorly understood to data. Hence, the aim of this study was to determine the pro-tumor function and mechanism of CUL4B in TCL.

Methods

The paraffin-embedded lymph node samples were collected from 30 newly diagnosed cases of T-cell lymphoma and 20 normal lymph nodes with approval in our hospital. TCL samples and purified normal T cells (including CD4+, CD8+, HLA-DR+, and HLA-DR-cells) (Pier Paolo Piccaluga et al., 2007) were evaluated for CUL4B expression. Peripheral blood mononuclear cells (PBMCs) were isolated from healthy donors. Expression levels of CUL4B mRNA and protein in TCL cells was detected by RT-PCR and western blot assay. Lentiviral vectors were used to transfected TCL cells to stably silence CULB. To evaluate the effects of CULB on cell proliferation and cycle, cell counting kit-8 and Propidium iodide staining were assessed respectively.

Results

We found that the expression levels of CUL4B were elevated in TCL samples compared with purified normal T cells via analyzing previously published gene expression data (Pier Paolo Piccaluga et al., 2007) (Fig. 1A). IHC assay indicated that CUL4B was higher expressed compared to normal lymph nodes (Fig. 1B). Moreover, RT-PCR and western blotting assays confirmed the elevated expression level of CUL4B in TCL cells compared with PBMCs in agreement with ICH assay (Fig. 1C-D). The function of cell viability maintenance of CUL4B as well as correlation with poor outcome has been reported previously in solid tumor. In consistent with previous studies in solid malignancies, knockdown CUL4B in TCL cells inhibited cell proliferation, induced G1/S phase arrest (Fig. 1E). Additionally, loss-of-function assay showed that the expression of phosphorylated AKT were reduced in TCL cells. Besides, p21 and p27 were observed in TCL cells (Fig. 1F).

Conclusion

Our investigations firstly identified the elevated expression levels and the oncogenic role of CUL4B in TCL tumorigenesis. Expression of CUL4B was upregulated in TCL. Targeting CUL4B with shRNA interference exerted therapeutic potential in abrogating cell survival, inducing cell cycle arrest through regulating the AKT signaling pathway. Taken together, targeting the expression of CUL4B may provide a novel approach to TCL therapy.

Disclosures

No relevant conflicts of interest to declare.
Toxicity Profile of Brentuximab Vedotin in Combination with Chemotherapy for Newly Diagnosed Patients with ALK+ ALCL: A Children's Oncology Group Study ANHL12P1
Authors: Lowe Eric J , Reilly Anne , Bollard Catherine M. , Barkauskas Donald A , Buxton Allen B , Hall David , Saguilig Lauren , Sandlund John T.
Published: October 2019

Background: Despite numerous treatment strategies, failure rates remain 25-30% for pediatric ALCL. Brentuximab vedotin is an antibody-drug conjugate containing an anti-CD30 monoclonal antibody linked to a tubulin inhibitor (monomethylauristatin E) which has significant activity as single agent in relapsed ALCL. The primary aim of ANHL12P1 (NCT01979536) is to determine the toxicity and efficacy of the addition of two novel agents (brentuximab vedotin or crizotinib) to standard chemotherapy (best arm of ALCL99) in children with newly diagnosed, non-localized, ALK+/CD30+ ALCL.

Methods: Patients on Arm BV received standard chemotherapy plus brentuximab vedotin. All patients initially received a 5 day prophase followed by 6 cycles of chemotherapy (dexamethasone, ifosfamide, cyclophosphamide, doxorubicin, etoposide, cytarabine, methotrexate) at 21 day intervals. Brentuximab vedotin was given on day 1 of each of the 6 cycles excluding prophase.

Results: From November 2013 to January 2017, 68 children with ALCL were enrolled on Arm BV. One patient was taken off protocol at the treating physician's discretion during prophase before receiving brentuximab vedotin leaving 67 eligible patients for toxicity evaluation. 66 of 67 patients completed all 6 cycles of chemotherapy resulting in 399 cycles evaluable. There were no toxic deaths, no cases of progressive multifocal leukoencephalopathy syndrome, and no cases of grade 3 or 4 neuropathy. Grade 3 or 4 toxicities which occurred in more than 5% of cycles included hematological toxicities, mucositis, and febrile neutropenia (see table). The mean/median interval between each cycle was: 22.7/22 days (cycle 1 to 2), 22.7/21 (2 to 3), 22.8/21 (3 to 4), 23/21 (4 to 5), and 22.8/21 (5 to 6). Interval between cycles was >28 days in only 3.6% of all cycles.

Conclusions: Arm BV of ANHL12P1 demonstrated that the addition of brentunixmab vedotin to standard chemotherapy does not cause significantly added toxicity. Brentuximab vedotin did not change the desired interval between cycles with the interval remaining similar from first to last cycle. It remains to be seen if the addition of brenutuximab vedotin improves overall or event-free survival compared to historical controls.

Table. Table.

Disclosures

Bollard: Cellectis: Honoraria, Membership on an entity's Board of Directors or advisory committees; Torque: Honoraria, Membership on an entity's Board of Directors or advisory committees; Neximmune: Honoraria, Membership on an entity's Board of Directors or advisory committees.
Enhanced Zebrafish Xenograft Platform Improves Hematopoietic Stem Cell Engraftment and Leukemogenesis
Authors: Rajan Vinothkumar , Melong Nicole , King Benjamin , Campbell Clinton , Berman Jason
Published: October 2019

The hallmark of hematopoietic stem cells (HSCs) is their ability to self-renew and differentiate into multiple blood cell types. Transplantation of human HSCs into immunocompromised mice is the gold standard for evaluating functionality and has been adapted to study leukemia initiating cells (LICs). However, specific cellular subsets do not survive post-xenotransplantation, due to a dependence on factors that may be missing in a non-primate microenvironment.

We have been optimizing the zebrafish as a xenograft platform. We created novel zebrafish that express human stem cell factor (SCF/KITLG), granulocyte-macrophage colony stimulating factor (GM-CSF/CSF2) and stromal cell-derived factor 1 alpha (SDF1α/CXCL12) - factors essential for survival and expansion of HSCs.

CMK (Down Syndrome- acute myeloid leukemia (AML)) and Jurkat (T-cell acute lymphoblastic leukemia) cell lines injected into the yolk sac of these multi-cytokine zebrafish larvae exhibit increased proliferation and migration to hematopoietic niches, namely the caudal hematopoietic tissue (=fetal liver) at 7 days post-injection (dpi) and the kidney marrow at 11 dpi, compared to injections in control casper larvae.

Patient-derived AML bone marrow samples (n=3) injected into the circulation of these multi-cytokine fish similarly displayed enhanced proliferation and migration with associated larval mortality from increased disease burden. Historically, human HSCs do not survive beyond 24 hpi in zebrafish xenografts (Pruvot et al. 2011). Remarkably, human umbilical cord blood-derived HSCs injected into the circulation of zebrafish larvae survived past three days-post-injection in multi-cytokine fish but not in casper controls.

Transgenic zebrafish expressing human cytokines provide an enhanced model for studying human leukemia and HSC biology in a system that more closely recapitulates the human microenvironment. By employing these humanized zebrafish and exploiting the inherent capacity of this small organism for rapid chemical screening, we have created a new preclinical paradigm for testing anti-leukemic therapies and HSC enhancing agents.

Disclosures

No relevant conflicts of interest to declare.
A Novel Enzyme Capture - ELISA for the Direct Determination of Factor XIII Activity
Authors: Durda Michael Andrew , Waller Amanda P. , Davis Kyle A , Kerlin Bryce A.
Published: October 2019

Background: Activated factor XIII (fXIII) mechanically stabilizes fibrin clots by catalyzing the formation of intramolecular crosslinks and retards plasmin-mediated fibrinolysis by incorporating intermolecular α2-antiplasmin (α2AP):fibrin crosslinks. High quality quantitative fXIII activity assays are required for the accurate and timely diagnosis of fXIII deficiency. Currently available high-throughput assays indirectly measure the transglutaminase activity of fXIII via detection of ammonia that is elaborated upon formation of ε-N-(γ-glutamyl)-lysyl crosslinking bonds. Unfortunately, these assays can be confounded by fXIII-independent ammonia producing and consuming reactions present in plasma. We thus set out to develop an assay which directly quantifies the transglutaminase activity. We hypothesized that fXIII could be captured in a microtiter plate with anti-fibrinogen antibodies and, following an activation step, its activity determined in a kinetic fashion via the rate of α2AP incorporation (measured in a subsequent ELISA step).

Methods: Co-immunoprecipitation and western blotting techniques were utilized to confirm fXIII/fibrinogen interactions and to determine the presence of basal interactions between fibrinogen and α2AP. Simultaneously, this approach enabled the determination of suitable antibody pairs for use in the enzyme capture-ELISA (EC-ELISA). In microtiter plates, fibrinogen-captured fXIII was activated with excess thrombin, physiologic calcium, and GPRP peptide (to block fibrin polymerization) in the presence of excess α2AP. FXIII activity was stopped with a potent fXIII-inhibitor (iodoacetamide) at predetermined time points. Subsequently, α2AP incorporation was determined with a primary anti-α2AP antibody and secondary HRP-conjugated antibody.

Results: Co-immunoprecipitation and western blotting experiments confirmed that both fXIII and α2AP circulate in complex with fibrinogen, as previously reported by others. However, α2AP incorporation significantly increased following incubation with activated factor XIII. Using the EC-ELISA method, we were able to demonstrate that α2AP is incorporated in a manner consistent with first order kinetics.

Discussion: These experiments demonstrate the feasibility of an EC-ELISA assay to directly measure fXIII activity. Further work will be required to determine the sensitivity of this assay in test plasmas with varying known quantities of fXIII and how its performance characteristics compare to currently available indirect activity assays. With further optimization, the EC-ELISA method may lead to an improved, high throughput diagnostic assay for clinical use.

Disclosures

No relevant conflicts of interest to declare.
Aspirin Use in Venous Thromboembolism Prophylaxis Following Hip or Knee Arthroplasty: A Network Meta-Analysis of Randomized Controlled Trials
Authors: Haykal Tarek , Kheiri Babikir , Barbarawi Mahmoud , Zayed Yazan , Shah Miran Muhammad , Chahine Adam , Abdalla Ahmed , Katato Khalil , Bachuwa Ghassan , Hadid Tarik , Kafri Zyad
Published: October 2019

Background: Patients undergoing hip or knee arthroplasty are at high risk of developing venous thromboembolism (VTE), such as deep-vein thrombosis and/or pulmonary embolism. Various thromboprophylactic strategies have been studied for the prevention of VTE in this population with different outcomes. Therefore, we aimed to evaluate the efficacy and safety of aspirin prophylaxis when compared with a placebo or anticoagulants in reducing such risk.

Methods: A comprehensive electronic database search in PubMed, Embase, the Cochrane Collaboration Central Register of Controlled Trials, and conference proceedings, was conducted for all randomized clinical trials (RCTs) comparing the clinical outcomes of aspirin versus anticoagulants or placebo for the prevention of VTE after hip or knee arthroplasty. The primary outcome was VTE. Secondary outcomes were mortality, major bleeding, and any bleeding. We calculated risk ratios (RRs) and 95% confidence intervals (CIs) using a random-effects model at the longest follow-up.

Results: We included 13 RCTs with 20115 total patients, mean age of 67.15±9.54 and a male percentage of 24.39%. Aspirin was associated with a non-significantly reduced VTE risk compared with other thromboprophylactic strategies (RR 0.87; 95% CI: 0.61-1.23; P=0.43). Compared with a placebo, aspirin was associated with significant reduction of VTE (RR 0.65; 95% CI: 0.47-0.89; P=0.008). There were no significant differences in the clinical outcomes between all groups with regard to mortality (RR 0.98; 95% CI: 0.86-1.11; P=0.72), major bleeding (RR 0.96; 95% CI: 0.50-1.84; P=0.91), or any bleeding (RR: 1.09; 95% CI: 0.82-1.44; P=0.56).

Conclusion: Among patients who underwent hip or knee arthroplasty, aspirin prophylaxis was associated with similar efficacy and safety outcomes when compared with anticoagulants. Compared with a placebo, aspirin prophylaxis was associated with significantly reduced VTE risk and a similar safety profile.

Table Table.

Disclosures

No relevant conflicts of interest to declare.
Imatinib Mesylate Is Safe and Effective in Maintaining Cytogenetic and Molecular Response Achieved with 1 Year Nilotinib First Line in Newly Diagnosed Chronic Myeloid Leukemia: Preliminary Results of a Prospective Clinical Trial
Authors: Moukalled Nour , Massoud Radwan , Mahfouz Rami , Elcheikh Jean , Bazarbachi Ali
Published: October 2019

Background: Achieving complete cytogenetic response (CCyR) at 12 months for patients with chronic phase chronic myeloid leukemia (CML) treated with imatinib as first line, is associated with significantly improved progression-free survival. Nilotinib has shown a faster and higher rate of CCyR and molecular response as compared to imatinib. However, nilotinib use is associated with diet restriction, higher financial costs, and its long-term use is incriminated in cardiovascular and metabolic complications.

Methods: In this prospective single center trial, we evaluated the ability of imatinib to maintain a CCyR achieved after 12 months of first-line treatment with nilotinib. Inclusion criteria were adult patients with previously untreated Philadelphia -positive CML in chronic phase, with a WHO performance status ≤ 2. Patients received 1-year treatment with nilotinib (300mg twice daily) then were shifted to imatinib 400mg daily.

Results: Eleven patients (5 females) were so far enrolled in the study, with a median age at diagnosis of 50 years (range 31-83). Patients were started on Nilotinib at a median of 29 days (range 2-32) from diagnosis. One patient had to discontinue nilotinib after 6 months and start dasatinib due to recurrent pancreatitis that resolved upon interruption of nilotinib. Eight patients completed one year of nilotinib, and were switched to Imatinib. The remaining 2 patients are currently on nilotinib as per protocol (less than 12 months since inclusion). Three patients on imatinib had to be switched back to nilotinib, because of imatinib intolerance (n=2; elevated liver transaminases and myositis) or because of loss of MMR (n=1). Therefore, at last follow up (median 64 months), 5 patients are on imatinib, 5 on nilotinib and 1 on dasatinib. All patients (100%) achieved complete hematological response (CHR) and CCyR at 3 and 18 months respectively, and maintained them thereafter. At 12 months, 6 out of 8 eligible patients achieved complete molecular response (CMR; n=3) or major molecular response (MMR; n=3). At 36 months, all eligible patients (n=7) were either in CMR (n=2) or in MMR (n=5). All patients who reached 4 years (n=6), 5 years (n=5) or 6 years (n=3) of follow up remained in either MMR or CMR. No patient developed long-term serious adverse event including cardiovascular or metabolic complications.

Conclusion: These findings suggest that imatinib can maintain MMR achieved on short-term nilotinib therapy. This strategy is a potentially safe and effective long-term treatment approach, minimizing costs and cardiovascular and metabolic complications. This however, needs confirmation with a larger number of patients.

Table. Table.

Disclosures

No relevant conflicts of interest to declare.
Efficacy of Newer Combinations and Fibril-Targeting for AL Amyloidosis: A Systematic Review of Transplant and Innovative Non-Transplant Therapies
Authors: Pandya Shaunak , Nunes Cavalcante Parr Nadia Carenina , Sardar Muhammad , Chiu Christine , Serafini Maria , Anwer Faiz
Published: October 2019

Background

Light-chain (AL) amyloidosis is a multi-organ amyloid deposition disease caused by misfolded protein aggregation. Current treatments have improved overall (OS) and progression-free survival (PFS) but challenges remain in improving therapy with newer agents and targeting amyloid protein deposits with novel immunotherapy. We review the literature regarding efficacy of existing chemotherapy in AL amyloidosis and summarize non-FDA approved novel drugs and monoclonal antibodies (mAbs) in early phase clinical development.

Methods

We searched databases including Cochrane library, PubMed and ClinicalTrials.gov for all prospective and retrospective studies (As of 4/15/2018) with measured hematologic response rate (HR) in patients with AL amyloidosis since 2000. Inclusion criteria included all prospective and retrospective studies with melphalan-based treatments, bortezomib combinations including bortezomib, cyclophosphamide and dexamethasone (VCD), bortezomib and dexamethasone (VD) and immunotherapies. We included all studies with at least 5 or more patients and reported HR.

Results

From 918 studies, we selected 57 studies (2640 patients) evaluating HR with melphalan-based stem cell transplant (SCT) treatments and non-stem cell transplant treatments including melphalan and bortezomib combinations. Other agents included daratumumab (anti-CD38 mAb), and ixazomib (proteasome inhibitor). Mean aggregate HR reported from studies with melphalan-based SCT treatment (17 studies, n=587) was 67%. Mean aggregate HR from all non-transplant treatments (40 studies, n=2053) was 64%. Of the non-transplant treatments, HR for melphalan-based treatments (21 studies, n=1148) was 59% and varied as follows: melphalan + lenalidomide + dexamethasone (57%) and melphalan + dexamethasone (52%). Mean aggregated HR for non-transplant bortezomib-based treatment (17 studies, n=859) was 72% consisting of VD (69%) and VCD (76%). HR with Ixazomib (1 study) and Daratumumab (1 study) was 52% and 76% respectively. Other novel drugs currently being studied include 11-1F4 (chimeric fibril-reactive mAb), GSK2398852 and GSK2315698 (anti-serum amyloid protein mAbs), and NEOD001 (anti-circulating soluble and deposited aggregated amyloid mAb).

Conclusion

For AL amyloidosis, melphalan-based SCT has shown effectiveness while VD and VCD demonstrate effectiveness in non-transplant patients. Further studies are warranted to evaluate novel proteasome inhibitors (Ixazomib) and emerging immunotherapy with daratumumab. Current trials including amyloid protein and fibril targeting (circulating and tissue-fixed) with novel immunotherapy are innovative and may have higher clinical efficacy, but need further testing.

Disclosures

No relevant conflicts of interest to declare.
Optimal Hemoglobin Level for Pediatric Sickle Cell Patients Who Undergo Adenotonsillectomy
Authors: Hu Zhongbo , Chang Simone Megan , Younis Ramzi , Alvarez Ofelia A.
Published: October 2019

Background:The prevalence of obstructive sleep apnea syndrome (OSA) in children with sickle cell disease (SCD) is higher than in the general pediatric population. Adenotonsillectomy involves significantly increased risks in patients with SCD because of the need for general anesthesia,which could induce a vaso-occlusive event. Various preoperative regimens have been suggested to reduce peri-operative risks for SCD patients.

Objective: We retrospectively reviewed the perioperative management for pediatric SCD patients undergoing adenotonsillectomy. We sought to identify the optimal preoperative hemoglobin (Hb) level for patients to have lowest risk of post-operative complications.

Methods: Children between 1 and 18 years old with SCD including HbSS, HbSC, HbS beta thalassemia (Sβ0 and Sβ+) who underwent adenotonsillectomy from 2007 to 2017 were identified at Holtz Children's Hospital. Patients without these diseases who underwent the procedures were collected as a control. The study was approved by the University of Miami Institutional Review Board (IRB). De-identified data were exported to GraphPad Prism version 5.02 to perform the statistical analyses. Means were analyzed by student t test, rates by Chi-squared test. p < 0.05 is considered statistically significant.

Results: Thirty-four patients with SCD (mean 7.1 yrs) and 145 controls (mean 7.0 yrs) were identified with adenotonsillectomy (see Table 1). SCD patients had significant longer hospital stay (2.6 versus 0.9 days, p< 0.001), and higher postoperative complication rates than the control group (20.6% vs 4.0%, p= 0.02). Most of the sickle cell patients' complications were hypoxemia (5 out of 7), except one supraventricular tachycardia and one headache. Four out of 7 patients with pretransfusion (pretrx) Hb level above 10 g/dL received manual exchange transfusion to keep Hb over 10 g/dL. None had complication or PICU stay. When patients' pretrx Hb levels was 9-10 g/dL, 5/8 received transfusion without significant complications, with the goal of not increasing post transfusion Hb over 11.5 g/dL. Most patients with pretrx Hb level 7-9 g/dL received simple blood transfusion. For patients with preoperative Hb level 9-11.5 g/dL, the PICU transfer rate was significantly lower than ones with Hb <9 or >= 11.5 g/dL (11.5% vs 60%, p=0.04; see Figure 1). Complication rates were also lower in the Hb 9-11.5 g/dL group, but without statistical difference (19.2% vs 40%, p=0.3). Data is limited by small sample size in the SCD group.

Conclusion(s): We concluded that for pediatric SCD patients planning for adenotonsillectomy, it is better to keep the posttransfusion Hb level at least above 9 mg/dL but less than 11.5 g/dL before procedure to avoid significant post operational complications. If pretransfusion Hb level is above 9 mg/dL, exchange transfusion by partial phlebotomy or simple transfusion (not to exceed 11.5 g/dL) may be considered.

Disclosures

No relevant conflicts of interest to declare.
Stat3 Regulates Alloreactive T Cell Susceptibility Towards PD-L1-Mediated Tolerance Signaling, Gvhd and GVL Activity
Authors: Yang Shijie , Wang Xiaoqi , Zhang Yuankun , Zeng Deye , Song Qingxiao , Martin Paul J , Zhang Xi , Zeng Defu
Published: October 2019

Prevention of graft versus host disease (GVHD) while preserving graft-versus-leukemia/lymphoma (GVL) activity of alloreactive donor T cells remains an elusive and long-sought goal. We and others reported that Stat3 deficiency in donor T cells results in expansion of Foxp3+ Treg cells and reduction of pathogenic Th17 cells, leading to prevention of chronic GVHD. We have now tested whether Stat3 deficiency in donor T cells can prevent acute GVHD while preserving GVL activity.

We transplanted wild-type (WT) or Stat3-/- T cells from C57BL/6 (H-2b) donors into irradiated WT BALB/c recipients bearing B cell lymphoma line BCL1 or aggressive acute lymphocytic leukemia (ALL) cells. WT donor T cells caused acute GVHD, as expected. Stat3-/- donor T cells did not cause acute GVHD but still had strong GVL activity. This separation of GVL activity from GVHD was associated with augmented expansion and impaired tolerance by Stat3-/- T cells in lymphoid tissues such as the spleen. In contrast, tolerance by Stat3-/- T cells was enhanced in GVHD target tissues by different mechanisms. The yield of Stat3-/- T cells in gut tissues was lower than the yield of WT T cells due to the markedly higher apoptosis of Stat3-/- T cells compared to WT T cells. The yield of Stat3-/- T cells in the lung and liver was not lower than the yield of WT T cells, but expression of anergy/exhaustion markers such as PD-1 and KLRG1 was markedly higher in Stat3-/- T cells than in WT T cells.

We recently reported that GVHD tissue expression of PD-L1 can tolerize donor CD8+ T cells in the absence of donor CD4+ T cells, but not when CD4+ and CD8+ T cells were given together. In contrast, PD-L1 expression in recipient GVHD target tissues tolerized Stat3-/- donor T cells when both CD4+ T cells and CD8+ T cells were given together. The induction of tolerance in Stat3-/- T cells by GVHD target tissue expression of PD-L1 depended on PD-L1/PD-1 interaction, which led to marked reduction of glycolysis in Stat3-/- T cells but not in WT T cells. These observations fit with previous findings that enhanced glycolysis was associated with enhanced GVHD activity of alloreactive T cells, as reported by Xue-zhong Yu's group. Our results indicate that Stat3 deficiency in donor T cells can augment their susceptibility to tissue PD-L1-mediated tolerance mechanisms such as down-regulation of glycolysis. Current experiments are testing whether Stat3 knockdown in mature T cells in the graft can prevent GVHD while preserving GVL activity. (Grant support: R01AI066008 to Zeng)

Disclosures

No relevant conflicts of interest to declare.
Psychological, Physical and Functional Importance of Patient Reported Outcomes Among Cancer Outpatients
Authors: Shrotriya Shiva , Walsh Declan , Sharma Mukta , Rayamajhi Supratik
Published: October 2019

Introduction:

There has been increased interest in patient-reported outcomes (PROs) as they allow consistent monitoring of patients' symptoms and quality of life. We evaluated the prevalence of PROs among cancer outpatients. Our objective was to look at the association of ET components with demographic and disease characteristics.

Methods:

PROs were prospectively collected cross sectional data which were later mixed with retrospectively obtained clinical data from the EMR. Predictors of ET components were determined by linear regression analysis. The predictors of those who required Help was determined by logistic regression.

Results:

The mean age was 58 (±14) years. 54% were male. The common cancer diagnoses were breast 17%, hematological (Leukemia) 17% and lymphoma 12%. Only a minority (7%) of participants required help (N = 1076) with the tablet-based surveys. Age categories ≤ 30 and 51-70 scored lower in mean depression ET. African Americans had a higher mean ET Anger scale. African Americans scored higher in mean need of Help -ET. Age 31-50 years was at 13% lower risk and 51-70 years was at 72% lower risk. Those with self-care problems were at higher risk of requiring Help with tablet computer.

Conclusions:

Many cancer patients visiting the cancer center had high pain/discomfort, anxiety/depression, problems with usual activities and mobility problems. Younger patients (31-50 years) had more distress and depression (ET). African Americans reported more problems with mobility, self-care, usual activities, pain/discomfort and anxiety/depression (EQ5D). Those with self-care problems were at risk to require help with tablet-based surveys.

Disclosures

No relevant conflicts of interest to declare.
When to Suspect Systemic Mastocytosis?
Authors: Kaplan Sibel , Kuskonmaz Baris , Soyer özge Ulusoy , Aytaç Selin , Gümrük Fatma
Published: October 2019

Background : Mastocytosis is a rare disease in children and it usually resolves by adolesence.However; it causes anxiety in parents and should be taken into consideration in patients with cutaneous eritematous lesions.

Aim: To find out the systemic mastocytosis patients who apply to pediatric hematology , pediatric allergy and dermatology policlinics.

Material and Methods: The files of 24 patients diagnosed with mastocytosis between January 2018 and August 2018 in our clinic were retrospectively analyzed.

Results: All patients had cutaneous mastocytosis, 14 patients had urticaria pigmentosa and 9 patients had solitary mastocytoma and one patient had diffuse cutaneous mastocytosis. The male-female ratio was: 2/1. The median age for onset of disease was 7,2 (0-48 months )months. The patients had different types of skin findings such as macules (n=16), plaques(n=6), papules(n=4) and bullae(n=3). None of the patients had anaphylaxis but two patients had angioedema. The diagnosis was confirmed with skin biopsy in 8 patients. The other were diagnosed due to typical history and physical examination.

None of the patients had cytopenia, hepatosplenomegaly.Only one patient triptase level >20 ug/l. The bone marrow aspiration and biopsy was done to 3 patients and all were normal.The patients were treated with H2 antihistaminics, local moisturizing creams and topical corticosteroid drugs in case of undesirable pruritus.

Conclusions: Most cases of childhood mastocytosis are observed in the form of cutaneous mastocytosis and ıt usually resolves by adolescence. If the patients have cytopenia, hepatosplenomagaly or triptase level >20 ug/l the bone marrow aspiration and biopsy is recommended.

Disclosures

No relevant conflicts of interest to declare.
Treatment of Unresectable Unicentric Castleman Disease with Therapeutic Embolization
Authors: Mohan Meera , Meek James C , Meek Mary Elizabeth , Broadwater Ralph Lynn , Stone Katie , Greenway Amy , Alapat Daisy , van Rhee Frits
Published: October 2019

Introduction: Unicentric Castleman disease (UCD) is a rare non-clonal lymphoproliferative disorder affecting one lymph node station. UCD can be an incidental finding on radiologic studies, whilst other patients have symptomatology due to compression of vital structures. Surgical extirpation is the preferred therapy and is usually curative, but unresectable UCD can represent a therapeutic challenge. Castleman lymph nodes are often highly vascularized, which offers the opportunity for therapeutic embolization. We report a series of 6 patients with unresectable UCD who were treated with embolization either as sole therapy or supplemented by cryoablation and surgery.

Methods: CT rotational angiography was performed to localize the arterial supply of UCD masses. Feeding vessels were selectively embolized using a 50:50 mixture of lipiodol and alcohol or 300-500 micron embospheres. A second arteriography was performed 2 to 3 months later to identify and embolize any new arterial channels.

Results: Data is summarized in Table 1. Of a cohort of 47 patients with UCD, 6 (13%) were found to have symptomatic, unresectable disease. All patients were HIV and human herpesvirus-8 negative. The median patient age was 34 years (range: 28-34); five patients were male and one was female. Disease was localized to the pelvis (n=3), mediastinum (n=2), and axilla (n=1). In all but one case, the histology was of the hyaline vascular variety. Four patients had failed R-CHOP, rituximab/steroids, or both. In 2 patients, embolization was done as primary therapy, while 3 underwent additional surgery. In 5 patients, embolization was performed twice to ablate secondary arterial channels that had appeared after the first procedure. Adjunctive cryoablation at the time of embolization was applied in 2 patients. All treated patients had major reduction in their lymph node mass. The median reduction in tumor bulk was from 274cc (range:13-969) to 21cc (range: 3-394). One patient with an axillary mass involving the brachial plexus failed therapy and received radiation. A second patient had regrowth of the UCD and responded to combination lenalidomide and obinituzumab. Responses were sustained for at least 2 years in the remaining patients.

Conclusion: A small number of case reports have been described UCD patients treated with arterial embolization as an immediate preoperative adjunct to surgery to limit intraoperative bleeding. In the present series, we utilized embolic devascularization to achieve cytoreduction rather than merely prevent surgical hemorrhage. Embolization was complemented by cryoablation and rendered surgery feasible. This case series highlights that effective disease control can be obtained of unresectable UCD using a multimodality approach in which vascular embolization plays a crucial role.

Disclosures

No relevant conflicts of interest to declare.
The Efficacy of Ultrasound Joint Examination for the Management in Patients with Hemophilia
Authors: Nishimura Shiho , Tomioka Keita , Shimomura Maiko , Mizoguchi Yoko , Sasaki Shinobu , Tani Chihiro , Matsubara Yoshiko , Awai Kazuo , Nakashima Yuko , Adachi Nobuo , Kobayashi Masao
Published: October 2019

The regular prophylactic replacement therapy with clotting factors has significantly reduced the arthropathy in patients with hemophilia. Early detection of the signs of joint damage is important to prevent the progressive joint damage. For the early detection, we have introduced the regular ultrasound (US) joint examination in patients with hemophilia from 2014 based on the hemophilia early arthropathy detection with ultrasound (Head-US) scanning protocol. US examination has been performed at the time of annual comprehensive clinic in Hiroshima University Hemophilia Center. Forty-five patients aged 1 to 33 years were enrolled and more than 280 joints were evaluated in this study. US joint examination was performed within 30 minutes without the sedation in all aged children. Abnormal findings, such as cartilage changes, subchondral bone erosion, synovial hypertrophy and/or joint effusion were observed in 8 out of 45 patients. The replacement therapy with clotting factors of all 8 patients aged 13 to 33 years was an insufficient prophylaxis during early childhood. The residual 37 patients aged less than 15 years have relatively followed the prophylactic treatment. The prophylactic regimen has been individually determined based on the trough levels of factor IX concentration in plasma for hemophilia B and pharmacokinetics of factor VIII using MyPKFitR for hemophilia A patients, respectively. The result suggests the importance of the careful prophylactic replacement therapy considering the factor concentration in plasma.

Four patients aged 4, 5, 18, and 19 years, respectively, suffered from painful and/or swollen joint episodes from 2014 to 2018. Urgent US examination without the use of any sedatives revealed ankle, knee, or hip joint bleeding in each case. According to the replacement therapy for breakthrough bleeding, hemoarthropathy was accessed by consecutive examination of US. These results suggest that US joint examination could be necessary for the early detection of joint damage and for appropriate treatment of hemoarthropathy. Collectively, utilization of US examination would be essential for the management through the life in patients with hemophilia.

Disclosures

No relevant conflicts of interest to declare.
Clonal Plasma Cells in AL Amyloidosis Are Dependent on Anti-Apoptotic BCL-2 Family Proteins
Authors: Fraser Cameron , Sanchorawala Vaishali , Sarosiek Kristopher , Sarosiek Shayna
Published: October 2019

Background: Immunoglobulin light chain (AL) amyloidosis is characterized by the production of clonal serum free light chains, which misfold and accumulate in tissues causing life threatening organ dysfunction and ultimately death. The treatment of AL amyloidosis targets the underlying population of clonal plasma cells, but existing therapies are not curative and ineffectively control the disease in many patients. Recent data have shown tremendous success in targeting anti-apoptotic BCL-2 family proteins as a novel therapy in hematologic disorders due to alterations in apoptosis and the function of anti-apoptotic proteins in malignant cells. BH3 profiling, a quantitative and functional assay that measures apoptotic priming and dependence on anti-apoptotic BCL-2 family members, has been used to identify and target apoptotic dependencies in hematologic disorders. Novel inhibitors of anti-apoptotic proteins, referred to as BH3 mimetics, have not yet been explored in AL amyloidosis due to insufficient understanding of apoptotic dependencies in this disease.

Methods: To date, bone marrow aspirates have been collected from 44 patients with newly diagnosed or relapsed/refractory AL amyloidosis being evaluated at the Amyloidosis Center at Boston University. BH3 profiling was performed on clonal plasma cells to measure dependencies on anti-apoptotic BCL-2 family proteins. Clonal cells were also treated with BH3 mimetics in vitro, including BCL-2, BCL-xL, and MCL-1 inhibitors as single agents, as well as in combination with current standard therapies (bortezomib, ixazomib, lenalidomide, and pomalidomide).

Results: Of the 44 enrolled patients, 16 are female (36%) and 28 are male (64%). The median age is 70 years (range, 47 to 84). Six patients were treatment naïve and the remainder had previous or current treatment for AL amyloidosis. Data obtained with BH3 profiling demonstrated that clonal plasma cells exhibit strong dependencies on anti-apoptotic BCL-2 family proteins, which may be altered by concurrent treatment with standard therapies. In the majority of patients, clonal plasma cells are highly dependent on the anti-apoptotic protein MCL-1 and undergo apoptosis when treated with an MCL-1 inhibitor. Intriguingly, this dependence is altered by treatment with proteasome inhibitors: clonal plasma cells become highly dependent on BCL-2 and undergo apoptosis in response to co-treatment with bortezomib and the FDA-approved BCL-2 inhibitor venetoclax.

Conclusions: BH3 profiling can effectively measure apoptotic dependencies in clonal plasma cells derived from bone marrow aspirates in patients with AL amyloidosis. Dependencies on BCL-2 family proteins (particularly MCL-1) are strong, but some variability between patients was observed, especially in combination with standard therapies. Biomarker-driven deployment of BH3 mimetics for treatment of AL amyloidosis would likely be highly effective.

Disclosures

No relevant conflicts of interest to declare.
Next Generation Flow for Multiple Myeloma Minimal Residual Disease: Igh Rearrangement NGS Is Complement to the NGF
Authors: Kim Sung-Min , Kim Jung-Ah , Jeong Dajeong , Yun Jiwon , Lim Kyu Min , Hwang Sang Mee , Yoon Sung-Soo , Lee Dong Soon
Published: October 2019

Background: Detection of leukemia-associated aberrant immuno-phenotype is used to assess minimal residual disease (MRD) by multi-parameter flow cytometry (MFC). However, detection of MRD by MFC remains to be a challenging due to the possible change in aberrant immunophenotype during disease progress. In our present study, we compared International Myeloma Working group (IMW) treatment response and NGF MRD, including BM PC% and cytogenetics. Thereon, we conducted IgH rearrangement study by NGS in cases showing discrepant results.

Methods: A total of 35 BM (35 myeloma patients at follow-up) was enrolled. We performed NGF using 8-color antibody panel using Navios flow cytometer and Infinicyt. Linearity of NGF was validated with myeloma cell line (U266) and BM specimen at initial diagnosis in myeloma patient. IgH rearrangement NGS was performed using Immunoseq assay (Adaptive Biotechnologies, Seattle, WA, USA). Paired specimen at initial diagnosis BM and follow-up BM were subjected to NGS study.

Results: Detection sensitivity of NGF was <0.001%. Patients who achieved CR or sCR showed MRD negativity in 63.6% (7/11). Twenty-three patients showed neoplastic PCs above LLOQ and their response criteria were 1 sCR, 3 CR, 2 VGPR, 3 PR, 1 MR, 5 SD, 3 progressive disease, 3 relapse, and 2 with unavailable response. Four patients who did not achieve CR (1 VGPR, 1 PR, 1 MR, and 1 SD) showed MRD negativity by NGF. In 4 patients with discrepancy between IMW treatment response and NGF, we compared the results of IgH NGS at initial BM with those after treatment. NGS revealed a persistence of residual clone in 1 patient and an acquisition of new clone after treatment. One patient had same dominant clone both initial diagnosis BM (95.2%; proportion of clone) and follow-up BM (45.8%). The other patient had newly appeared clones in follow-up BM (6.12%, 5.63%, 3.42%, 3.11%, 3.09%) which clones were absent in initial diagnosis BM. The other 2 patients showed heterogeneous clones without dominant clone at follow-up BM by NGS. Results of FISH and immunofixation are summarized in Table 1. This results show IgH rearrangement NGS can detect malignant clone that could not be identified by using NGF.

Conclusions: Thirty-six percent of patients who did not achieve CR showed NGF MRD negativity and NGS revealed residual clones in half of them. Switching of immunophenotypes of neoplastic PC can escape monitoring of NGF, and complementary NGS test is needed to catch such drifting clones for monitoring of MRD in MM.

Disclosures

No relevant conflicts of interest to declare.