bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pharmacokinetics and Biodistribution of Locked Nucleic Acid (LNA)-Mir-221 Inhibitor: A New Promising Anti-Myeloma Agent
Authors: Gallo Cantafio Maria Eugenia , Gulla Annamaria , Amodio Nicola , Leone Emanuela , Morelli Eugenio , Biamonte Lavinia , Neri Antonino , Tagliaferri Pierosandro , Tassone Pierfrancesco , Di Martino Maria Teresa 
Published: October 2019

Abstract
miR-221/222 are highly homologous microRNAs (miRNAs) whose upregulation has been found in several malignancies, including multiple myeloma (MM). Both miRNAs are thought to promote cell proliferation via down-regulation of p27 and/or p57, two negative regulators of G1 to S phase cell cycle progression.
We proved that silencing of both miRNAs results in significant anti-MM activity and in downregulation of canonical targets both in vitro and in vivo. In the aim to progress to clinical translation, we designed an original 13mer synthetic inhibitor, specific for systemic delivery, named LNA-i-miR-221, which took advantage from both locked nucleic acid (LNA) technology and phosphorothioate backbone, for increasing the seed sequence binding and nuclease resistance in vivo, respectively. Since no data are presently available, we evaluated the specificity of LNA-i-miR-221 to inhibit endogenous miR-221 and the pharmacokinetic properties (i.e. tissue distribution) in mice and Cynomolgus monkeys.
We demonstrated that LNA-i-miR-221 inhibit growth and survival of MM cells bearing high miR-221 levels. Moreover, LNA-i-miR-221-mediated silencing of miR-221 triggered upregulation of p27Kip1 mRNA and protein, evaluated by q-RT-PCR and western blotting, respectively. In vivo, i.p.treatment with 25 mg/kg of LNA-i-miR-221 reduced tumor growth in SCID/NOD mice bearing MM xenografts. Tumors and vital organs (including liver, kidney, bone marrow and heart) were harvested from treated animals and evaluated for pharmacokinetics purposes using in situ hybridization (ISH) assay. After a single i.p. dose of 25 mg/kg, we detected the presence of LNA-i-miR-221 from 2 up to 7 days in tumors and mouse tissues. Interestingly, no toxicity was observed even after long-lasting presence of the inhibitors in vital organs. We also evaluated the LNA-i-miR-221 half-life in mouse plasma using HPLC-MS/MS, which confirmed the rapid uptake of LNA-i-miR-221 in tissues. To evaluate the maximum tolerated doses, in vivo dose escalation treatments was performed using doses from 10 to 100 mg/kg delivered at days 1,4,8,15,22. All treatments were well tolerated. No changes in mice behavior or organ toxicity were observed in treated mice.
Finally, a pilot toxicity study was performed in Cynomolgus monkeys. PK results demonstrated that LNA-i-miR-221 has a short serum half-life with a rapid tissue uptake and minimum urinary escretion.
In conclusion, our results suggest that LNA-i-miR-221 is a promising anti-MM agent associated with a safety profile in mice and in monkeys, supporting the rationale for development of this novel miR-221 inhibitor in early clinical trials.

Disclosures
No relevant conflicts of interest to declare.
TLR5 Agonist Recombinant Flagellin, CBLB502 Increases Anti-CMV Immunity By Increasing T-Bet and Eomes Expressing T Cells after MCMV Infection
Authors: Hossain Mohammad Sohrab , Al-Kadhimi Zaid , Waller Edmund K 
Published: October 2019

Abstract
We have previously shown that highly purified cGMP grade recombinant flagellin, CBLB502, a TLR5 agonist, protected allo-HSCT recipients from GvHD without compromising anti-CMV immunity of donor T cells in vivo. A single prophylactic dose (25mg/mouse i.p) of CBLB502 in wild type C57BL/6 (B6) mice also enhanced anti-CMV immunity of NK cells against CMV. CMV is a potential opportunistic infectious agent and most often causes severe life-threatening clinical complications in immunocompromized patients, particularly in cancer patients treated with allogenic bone marrow transplantation (allo-BMT). A mechanism by which CBLB502 protected host against CMV infection has not been well investigated. Transcription factors ROR gammaT, T-bet and EOMES are known to control immune responses of innate and adaptive immunity in vivo. In this study, we have explored the transcription factors ROR gammaT, T-bet and EOMES expressing NK and T cells in CBLB502-treated and MCMV-infected B6 mice. Total numbers of spleen cells increased by 1.7-fold within 48 hours of CBLB502 treatment (day 0 MCMV infection) and by 2.3-fold on day 3 after MCMV infection compared with PBS-treated MCMV-infected mice. T-bet expressing CD8+ T cells (not CD4+ T cells) increased significantly on day 0 of mCMV infection, 48hrs after CBLB502 treatment (CD4, p=0.1; CD8, p<0.001) and both CD4+ and CD8+ T cells increased significantly on day 3 (CD4, p<0.001; CD8, p<0.01) after MCMV infection. CBLB502 treatment had no effect on the ROR gammaT expression on T cells. Interestingly, although EOMES expressing CD4+ and CD8+ T cells increased significantly on day 0 (48 hrs after CBLB502 treatment) (CD4, p<0.01; CD8, p<0.01) and day 3 (CD4, p<0.001; CD8, p<0.001) in the spleens of CBLB502-treated mice compared with the PBS-treated mice, EOMES expression on CMV-specific tetramer+ CD8+ T cells rapidly decreased by day 10 after infection in both CBLB502 and PBS treated mice. However, >90% of CMV-specific CD8+ T cells had persistent expression of T-bet even after 30 days post CMV infection. In contrast, CBLB502 treatment did not have a significant effect on the numbers of ROR gammaT, T-bet and EOMES expressing NK cells compared with PBS-treated mice. These data suggest that 1) CBLB502-induced anti-CMV immunity of T cells is persistently maintained through increased T-bet expression but not by EOMES expression; and 2) CBLB502-induced anti-viral immune regulation is not maintained through transcription factor ROR gammaT. 3) CBLB502-induced increased anti-CMV immunity of NK cells is independent of transcription factors ROR gammaT, T-bet and EOMES. Taken together, these data support the early peri-transplant use of TLR5 agonists as a novel method to enhance antigen-specific anti-viral immunity in recipients of allo-BMT.

Disclosures
No relevant conflicts of interest to declare.
Cytokine Gene Polymorphisms and Epstein-Barr Virus Infection after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Lin Ren , Fan Zhiping , Zhao Ke , Jiang Qianli , Sun Jing , Liu Qifa 
Published: October 2019

Abstract
Backgroud:
Epstein-Barr virus (EBV) infection/reactivation and associated diseases remain common complications in recipients of HSCT, leading to severe end-organ diseases and malignance. Single nucleotide polymorphism (SNP) in cytokine genes is considered to be related with EBV-associated post-transplant lymphoproliferative disorders (PTLD) in solid-organ transplantation recipients. In this study, we analyzed the association between cytokine gene polymorphisms and EBV infection/reactivation or diseases in recipients undergoing allo-HSCT.
Methods:
A total of 233 patients who received allo-HSCT between March 2012 to December 2014 were enrolled in this study. Ten SNPs, including IL-1β -511 rs16944, IL-1RN +11100 rs315952, IL-2 -330 rs2069762, IL-4 -590 rs2243250, IL-10 -592 rs1800872, IL-12 +1188 rs3212227, TNF-α -308 rs1800629, TGF-β1-509 rs1800469, TGF-β1 +869 rs1800470, IFN-γ +874 rs2430561, were tested. The SNPs genotypes in patients with or without EBV infection/reactivation and associated diseases were compared. Besides, the risk factors for EBV infection/reactivation were studied.
Results:
Seventy-four patients developed EBV infection/reactivation. The patients with EBV infection/reactivation had higher frequencies of donor IL-1β -511 TT genotype, donor IL-4 -590 TT genotype and recipient TNF-α -308 GG genotype than those without (p=0.021, p=0.004, p=0.020, respectively) while the frequencies of donor IL-1β -511 CC genotype, donor IL-1RN +11100 TT genotype, donor IL-2 -330 TT genotype, donor IL-4 -590 CC genotype and recipient TNF-α-308 GA genotype in patients with EBV infection/reactivation were lower than those without (p=0.041, p=0.029, p=0.005, p=0.011, p=0.042, respectively). Multivariate analysis showed donor IL-4 -590TT genotype (p=0.016, HR=1.907, 95% CI =1.130-3.218) and recipient TNF-α -308GG genotype (p=0.002, HR=3.550, 95% CI=1.613-7.812) were risk factors for post-transplant EBV infection/reactivation while donor IL-1RN +11100TT genotype (p=0.001, HR=0.382, 95% CI=0.218-0.670) was protective factors for post-transplant EBV infection/reactivation. Twenty-one patients developed EBV-associated diseases. The patients who developed EBV-associated diseases had higher frequency of donor IFN-γ +874 AT genotype than those did not (p=0.027). On the contrary, the frequencies of donor IL-1β-511 CC genotype, donor IL-10 -592 AA genotype, donor IL-12 +1188 AA genotype and donor IFN-γ +874 AA genotype in patients with EBV-associated diseases were lower than those without (p=0.019, p=0.018, p=0.018, p=0.010, respectively).
Conclusion:
Several SNPs in cytokine genes might be associated with EBV infection/reactivation and the development of EBV-associated diseases in recipients of allo-HSCT. However, these association should be studied further.

Disclosures
No relevant conflicts of interest to declare.
ASN002: A Potent Dual SYK/JAK Inhibitor Currently in a Phase I/II Study Shows Strong Antitumor Activity in Preclinical Studies
Authors: Rao Niranjan S. , Reddy Sanjeeva , Sandeep Gupta , Damle Nitin K , Aranapakam Venkatesan M , Thompson Scott , Smith Roger , Plasse Terry , Bristow Penelope , Tolcher Anthony , Papadopoulos Kyriakos P , Patnaik Amita , Rasco Drew W 
Published: October 2019

Abstract
ASN002 is an orally bioavailable potent dual inhibitor of spleen tyrosine kinase (SYK) and Janus kinases (JAK) with low nM IC50 values in biochemical assays. It is currently under clinical evaluation in a phase I/II study for safety, tolerability, pharmacokinetics, pharmacodynamics, and preliminary efficacy in diffuse large B-cell lymphoma (DLBCL), Mantle cell lymphoma (MCL) and Follicular lymphoma (FCL). SYK and JAK are tyrosine kinases that play important roles in various types of lymphomas, solid tumors, myeloproliferative and inflammation disorders. Inhibition of these targets has been shown to suppress tumor growth in various preclinical and clinical studies. ASN002 showed anti-proliferative activity in a broad panel of cell lines representing both solid and leukemia/lymphoma tumor types. ASN002 potently induced apoptosis in a DLBCL cell-line and showed a strong dose dependent tumor growth inhibition in multiple DLBCL xenograft models. ASN002 also showed strong anti-proliferative activity in an ibrutinib-resistant cell line. In mechanistic cell-based studies, ASN002 strongly suppressed IgE and IL-6 stimulated responses that are thought to be mediated by SYK and JAK, respectively. ASN002 exhibited good oral bioavailability and metabolic stability in animal models. ASN002 is being evaluated in patients with solid tumors and hematological malignancies (excluding those with extensive marrow involvement) in the dose escalation phase which employs a 3+3 open label design. Sequential doses of 10, 20, 30, 40 and 50 mg BID are planned. Patient accrual is continuing and no dose limiting toxicities were identified in the first cohort. Data on safety, tolerability, pharmacokinetics and pharmacodynamics will be presented.

Disclosures
Rao: Asana BioSciences: Employment, Equity Ownership. Reddy:Asana BioSciences, LLC: Employment, Equity Ownership. Sandeep:Asana BioSciences, LLC: Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees. Thompson:Asana BioSciences, LLC: Employment, Equity Ownership. Smith:Asana BioSciences, LLC: Employment, Equity Ownership. Plasse:Asana BioSciences, LLC: Consultancy. Bristow:Asana BioSciences, LLC: Consultancy. Tolcher:AbbVie: Consultancy; ArQule: Consultancy; Bayer: Consultancy; BioMed Valley Discoveries: Research Funding; Janssen R&D: Consultancy; Asana BioSciences, LLC: Consultancy, Research Funding. Patnaik:Asana BioSciences, LLC: Research Funding. Rasco:Asana BioSciences, LLC: Research Funding; Celgene: Research Funding.
Tyrosine Kinase Inhibitor Discontinuation in Chronic Myeloid Leukemia: Single US Center Experience
Authors: Khoury H Jean , Heffner Leonard T , Arellano Martha , Jillella Anand P , Kota Vamsi K , Winton Elliott F. 
Published: October 2019

Imatinib (IM) can be safely discontinued in patients with chronic myeloid leukemia (CML) with sustained complete molecular remission. Previous publications from France and Australia have shown that overall 40% maintain MMR or MR5 for up to 5 years after IM is stopped. We report single US center experience with tyrosine kinase inhibitor (TKI) discontinuation. Between 06/2010 and 7/2015, 22 patients with CML in chronic (CP, n=19), accelerated (AP, n=2) and lymphoid blast phase (LBP, n=1) discontinued IM (n=17), dasatinib (DAS, n=3) or bosutinib (BOS, n=2), and were monitored by qPCR for BCR-ABL1 monthly for the first 3 months, quarterly for the following 2 years and then bi-annually. TKI was restarted in case of confirmed loss of MMR on a repeat qPCR. Reason for TKI discontinuation was predominantly driven by patients' request and TKI intolerance. Median age was 66 (range, 21-84). The 3 who discontinued DAS had IM-resistant (loss of CCyR; n=1), IM-intolerant CP (n=1), or received DAS as first-line agent (n=1). BOS was discontinued for IM-intolerant CP (n=1), or while in CR2 in a patient with LBP that transformed from IM resistant CP and relapsed following chemotherapy (HCVAD). Median duration of TKI therapy pre-discontinuation for the entire cohort was 89 months (range, 26-106). Three patients are not evaluable due to short follow-up (TKI stopped between 5/2015 and 7/2015).
With a median follow-up of 40 months (range, 8-60), 7 (41%, 6 CP and 1 AP), all previously on IM lost MMR a median of 3 months (range, 3-24) after TKI was stopped and restarted IM. Loss of CHR occurred 13 months after loss of MMR in 1 patient who elected not to restart IM at the time MMR was lost, due to complications from cardiac transplant rejection. All 7 achieved MMR following restart of IM. Median duration of TKI therapy pre-discontinuation for these 7 patients was 60 months (range, 48-98). 12 patients (59%, 10 CP, 1 AP, 1 LBP) remain off TKI and have not lost MMR, 8 with continuously undetectable BCR-ABL1; and 4 had 1-2 transient detectable BCR-ABL1 at MR4 levels. Median duration of TKI therapy pre-discontinuation for these 12 patients was 87 months (range, 26-106). Loss of MMR-free survival is depicted in the Figure. We conclude that, similar to previous reports, TKI can be safely discontinued in patients with CML without reappearance of BCR-ABL1 in 50-60%.
Figure 1. Figure 1.

Disclosures
Jillella: Seattle Genetics, Inc.: Research Funding. Kota:Leukemia Lymphoma Society: Research Funding; Pfizer: Membership on an entity's Board of Directors or advisory committees.
Pharmacologic Modulation of STAT1 and Wnt Signaling in Chronic Lymphocytic Leukemia
Authors: Wu Christina , Lao Fitzgerald S , Choi Michael Y. , Carson Dennis A. 
Published: October 2019

Abstract
BACKGROUND: The STAT and Wnt signaling pathways play critical roles in early lymphocyte development and function, with considerable cross-talk between them. STAT1 has been reported to be constitutively phosphorylated at SER727, and often at TYR701, in CLL patients. STAT1 activation can regulate the function of the IRF8 transcription factor. Polymorphisms of the IRF8 gene have been associated with both familial and sporadic CLL, and can influence Wnt signaling. Hence, both STAT1 and Wnt are potential pharmacologic targets for CLL therapy, particularly in patients that are resistant to, or intolerant of BTK and PI3K inhibitors. We previously reported that the electrophilic drug dimethylfumarate (DMF, Tecfidera, Fumaderm) could inhibit Wnt signaling in CLL. Here we have investigated its effects on constitutive and inducible STAT1 signaling, and on IRF8 expression, in primary CLL cells.
METHODS: Primary leukemia cells from patients with CLL were cultured with DMF at pharmacologically relevant doses (3 uM to 30 uM) for 4 hours prior to analysis of STAT1 phosphorylation and IRF8 expression. We assessed the effect of DMF with and without pre-stimulation by lipopolysaccharide (LPS), Wnt3a, or a TLR7 agonist. To further evaluate the effect on CLL cells in a microenvironment that stimulates these pathways, we utilized a murine CLL xenograft model. We implanted primary leukemic cells from patients with CLL into the peritoneum of Rag2 deficient immune-compromised mice. Groups of mice were then treated with DMF at a dose of 10 mg/kg by oral gavage. CLL cells were retrieved 4 to 5 hours later by peritoneal lavage and analyzed.
RESULTS: We confirmed that both SER727 and TYR701 are phosphorylated in the majority of primary CLL cells from both VH mutated (high risk) and unmutated (low risk) clones, but not in normal peripheral blood mononuclear cells (PBMC). A 4 hours exposure of CLL cells to DMF inhibited TYR701 phosphorylation, but had no effect on SER727 phosphorylation nor on total STAT1 levels. We also confirmed that IRF8 levels are 6-8 fold elevated in CLL cells, compared to normal PBMC. IRF8 expression was inhibited by DMF treatment in 7 out of 11 CLL patients. Similar effects were observed in vivo. Wnt or LPS stimulation increased IRF8 levels in normal PBMC, but did not alter the already elevated IRF8 levels in CLL cells.
CONCLUSIONS: Activation of the STAT1/IRF8 pathway, like the Wnt pathway, is highly characteristic of CLL. DMF treatment of CLL cells can interrupt these signaling cascades, interfering with leukemia cell survival.

Disclosures
Choi: AbbVie: Consultancy, Other: Advisory Board, Research Funding; Gilead: Consultancy, Other: Advisory Board, Speakers Bureau.
Switching to Warfarin after 6-Month Completion of Anticoagulant Treatment for Cancer-Associated Thrombosis
Authors: Chai-Adisaksopha Chatree , Iorio Alfonso , Crowther Mark A. , de Miguel Javier , Salgado Estuardo , Zdraveska Marija , Monreal Manuel 
Published: October 2019

Abstract

Background: Low-molecular-weight heparin (LMWH) is considered to be an anticoagulation therapy for the treatment of cancer-associated thrombosis. The duration of treatment is recommended to maintain at least 3-6 months after the diagnosis. However, the data on continuing LMWH treatment beyond six months remains unclear.
Methods: Consecutive cancer-associated thrombosis patients who were enrolled in RIETE Registry were evaluated. We systematically selected the patients who completed treatment with LMWH for 6 months. The patients were divided into two groups whether they continued to receive LMWH or switched to warfarin. The main outcomes were recurrent venous thromboembolism (VTE), major bleeding and total bleeding. Survival curves were generated using Kaplan-Meier method and the curves were compared using the log-rank test. Hazard ratio (HR) with corresponding 95% confidence interval (CI) were calculated using Cox-proportional hazard (PH) model.
Outcomes: of the 1,502 eligible patients who completed 6-month anticoagulant therapy, 763 patients continued to received LMWH and 739 switched to warfarin. There was no significant difference in terms of recurrent VTE between two study groups (hazard ratio [HR] 0.67, 95% confidence interval [CI]; 0.44-1.02, p=0.06), Figure 1. The cumulative incidence of major bleeding was 2.6% in LMWH group and 2.7% in warfarin group (HR 1.05, 95%CI; 0.79-1.55, p=0.79), Figure 2. The cumulative incidence of total bleeding was 6.7% in LMWH group and 7.0% in warfarin group (HR 0.92, 95%CI; 0.62-1.37, p=0.70).
Conclusions: In patients with cancer-associated thrombosis who completed 6-month of anticoagulation therapy, switching to warfarin is not associated with increase in recurrent VTE, major bleeding or total bleeding when compared to continuing LMWH. Warfarin is an acceptable alternative anticoagulant in cancer-associated thrombosis patients who do not tolerate long-term treatment with LMWH.

Disclosures
Monreal: Bayer: Consultancy, Membership on an entity's Board of Directors or advisory committees; sanofi: Consultancy, Membership on an entity's Board of Directors or advisory committees; boehringer: Consultancy, Membership on an entity's Board of Directors or advisory committees; daichii: Consultancy, Membership on an entity's Board of Directors or advisory committees.
Pronostic Value of Pretransplant Echocardiography in Allogeneic Hematopoietic Cell Transplantation
Authors: Mauricio Sarmiento , Espigado Ildefonso , Parody Rocio , Marquez Francisco , Falantes Jose , Caballero Teresa , Calderon Cristina , Carmona Magdalena , Lopez Jose , Perez-Simon Jose A. 
Published: October 2019

Abstract

Several scores have been developed, such as the Sorror index, which allow to identify before transplant subgroups of patients with different risks of death after allogeneic transplantation (AloTH). One of the parameters included in all these scores and routinely used in the pretransplant evaluation include cardiac function studies. However, it has been heterogeneously defined mainly based according to the ejection fraction using different approaches such as MUGA and echocardiography. The aim of our study was to evaluate the prognosis impact of pretransplant echocardiographic findings.
We retrospectively analyzed a total of 276 medical records of patients undergoing AloTH between 2006 and 2014. Additional data regarding demographic parameters, underlying disease, donor type, conditioning, engraftment, acute and chronic graft versus host disease, overall survival and cause of death were also included into the analysis.
The mean ejection fraction was 61%, with 20 patients displaying an ejection fraction <50%. 19 patients (7%) had diastolic dysfunction, 5% valvular disease and 4% pericardial effusion. Of the 19 patients with diastolic dysfunction, 57% had been treated with anthracyclines: 7 patients (36%) were diagnosed with acute leukemia and 4 (21%)patients with lymphoma. In multivariate analysis we found that diastolic dysfunction was a predictor for survival (HR 5.43, 95% CI: 0,08-20,2, p = 0.028) and non-relapse mortality (HR = 5.7, 95% CI: 0,04-0,7, p = 0.002). Neither ejection fraction nor any other echocardiographic parameter predicted the risk of death.
In the present study pretransplant ejection fraction did not influence the prognosis of patients undergoing allogeneic transplantation; however, impaired diastolic function seems significantly influences post-transplant mortality. Therefore, in the pretransplant evaluation, diastolic function must be assessed in order to better define patient risks..
Figure 1. Figure 1. Figure 2. Figure 2.

Disclosures
Falantes: Celgene: Honoraria.