bluuebunny/crossref_metadata_2025_split huggingface dataset:
Efficacy and Safety of JWCAR029 in Adult Patients with Relapsed and Refractory B-Cell Non-Hodgkin Lymphoma
Authors: Yan Zixun , Wang Wen , Zheng Zhong , Hao Ming , Yang Su , Li James , Cheng Shu , Wang Li , Zhao Wei-li 
Published: October 2019

Abstract
Introduction
JWCAR029 is a novel CD19-directed 4-1BB stimulated chimeric antigen receptor T (CAR-T) cell type, which is different from JWCAR017 with independent production of CD4 and CD8 T cells and transfusion in non-fixed ratio. We conducted a single arm, open-label, dose escalation Phase I trial of JWCAR029 in relapsed and refractory B-cell non-Hodgkin lymphoma (NCT03355859).
Methods
From January to July 2018, 10 patients have been enrolled in this trial, including eight diffused large B cell lymphoma (DLBCL) and two MALT lymphoma, with median age of 47 years (range 32 to 59 years). All the patients received immunochemotherapy as induction and more than two lines of salvage treatment. Two patients received bridging chemotherapy after T-cell collection due to rapid tumor progression, followed by re-evaluation before CAR-T cell infusion. Lymphodepletion preconditioning was accomplished by fludarabine 25mg/m2/d and cyclophosphamide 250mg/m2/d on Day-4 to D-2, followed by CAR-T cell infusion on Day0. JWCAR029 was administrated as a single infusion in escalation dose levels, from 2.5×107 CAR-T cells (dose level 1, DL1) to 5.0×107 CAR-T cells (dose level 2, DL2) and to 1.0×108 CAR-T cells (dose level 3, DL3) according to mTPI-2 algorithm. Circulating blood count, serum biochemistry, and coagulation status were follow-up after infusion. Cytokines were assessed on a Luminex platform. Tumor evaluation was performed on Day 29 by PET-CT. PK data were detected by flow cytometry and real-time quantitative polymerase chain reaction system. All the adverse events were recorded. The study was approved by the Shanghai Rui Jin Hospital Review Board with informed consent obtained in accordance with the Declaration of Helsinki.
Results
The demographic characteristics of the patients were demonstrated in Table 1. Among six evaluable patients (3 of DL1 and 3 of DL2), the ORR was 100% on Day 29, including four complete remission and 2 partial remission. Cytokine release syndrome (CRS) was 100% in Gr 1, with main symptoms as fever (<39.0 degrees), fatigue, and muscle soreness. No neurotoxicity was observed. Four of the six patients with fever >38.0 degrees used prophylactic IL-6 Inhibitor (8mg/kg, ACTEMRA, two patients administered twice). No patients received steroids. The CRS showed no difference between dose level groups (p>0.99). Adverse effects included leukopenia (Gr 3-4: 83.3%, Gr 1-2: 16.7%), hypofibrinogenemia (Gr 1: 16.7%, Gr 2-4: 0%), liver dysfunction (Gr 1: 33.3%, Gr 2-4: 0%), elevated CRP (Gr 1: 83.3%, Gr 2-4: 0%), ferritin (Gr 1-2: 83.3%, Gr 2-4: 0%), or IL-6 (Gr 1-2:100%, Gr 3-4: 0%, Table 2).
Conclusion
Although long-term follow-up was needed, the preliminary data of six patients in this trial have demonstrated high response rates and safety of JWCAR029 in treating relapsed and refractory B-cell non-Hodgkin lymphoma.

Disclosures
Hao: JW Therapeutics: Employment, Equity Ownership.
A Successful Experience with Brentuximab Vedotin in Relapsing/Refractory Cutaneous T-Cell Lymphoma: Two Case Reports
Authors: Gorenkova Liliya , Kravchenko Sergey K. , Kovrigina Alla M. , Savchenko Valery G. 
Published: October 2019

Abstract
Primary cutaneous T-cell lymphomas are non-Hodgkin lymphomas of the skin defined by the presence of malignant T lymphocyte clonality.
Mycosis fungoides (MF) accounts for more than half of these lymphomas, while CD30+ lymphoproliferative skin diseases (primary cutaneous anaplastic lymphoma and lymphomatiod papulosis (LyP)) comprise 25% of these neoplasms, and rarely seen tumors including subtypes of primary cutaneous peripheral T cell lymphoma nonspecified (NOS).
A high expression of CD30 antigen on tumor cells is detected in primary cutaneous CD30+ lymphomas and transformed MF, whereas it may be observed in other entities, but a low level.
While most patients with cutaneous CD30+ lymphoproliferative disorders experience an indolent disease course with an excellent prognosis, up to 30% have progressive disease with 8% of patients succumbing to their cancer. Systemic immunomodulatory or chemotherapeutic agents are often used for patients with such advanced disease, with CD30 being an increasingly attractive therapeutic target.
Herein, we report two cases of a relapsing lymphomatiod papulosis and a refractory primary cutaneous peripheral T cell lymphoma NOS. Target therapy with brentuximab vedotin either as a single agent or combined with chemotherapy resulted in a stable long-term remission.
Patient 1, a man of 45 years of age, received treatment for Hodgkin lymphoma about 5 years ago. Already at that time, he noted self-regressing crops of pruritic papules or nodules on the face. In a year after chemotherapy had stopped, this skin elements recurrent with subsequent partial regression. LyP type D was identified on histological examination (picture 1).
Following several lines of therapy, frequent relapses were noted and a lesser quantity of lesions tended to resolve leaving scars and hyperpigmentation.
The complete remission was achieved after 6 cycles of brentuximab vedotin, at a follow-up period of 12 months no evidence of relapse were found (picture 2).
Patient 2, a 30-year aged man. First solitary skin lesions occurred 2.5 years ago. The patient received combination therapy with low doses of chemotherapeutic agents and subsequently had progression of the disease. The patient received more 3 lines of high-dose chemotherapy; however, the disease was rapidly progressive with increasing size of lesions and newly appearing tumor lesions in the skin (picture 3). The remission was achieved after a combination regimen which included Dexa Beam courses and brentuximab vedotin (picture 4). The matched allogeneic bone marrow transplantation is planned for the patient to consolidate the remission.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
The Role of DNAM-1/Tigit Axis of NK Cells in the Regulation of Alloreactive T Cell Responses and the Potential Mechanism
Authors: Sheng Lixia , Ouyang Guifang , Qitian MU , Huang He 
Published: October 2019

Abstract
Objective: Previous studies has showed the important role of NK cell in the regulation of alloreactive T cell response and negative regulation of GVHD. The paired receptors DNAM-1 and TIGIT, which bind the same ligands but have opposite effects on NK cell function, might present as potential targets for the GVHD treatment. This study is designed to explore the role of TIGIT/DNAM-1 balance of NK cells in the regulation of alloreactive T cell responses and the potential mechanism.
Methods: Antibodies blocking of TIGIT or DNAM-1, over-expression of TIGIT or DNAM-1 via lentiviral transduction and knockdown of TIGIT or DNAM-1 by lentiviral shRNA were used to manipulate the TIGIT/DNAM-1 balance on NK cell. Cytotoxicity assay using alloantigen activated T cells as targets were used to evaluate the regulating function of NK cell on alloreactive T cell responses. Western blot and small molecule inhibitors against PI3K were combined to investigate whether the PI3K-Akt-ERK signaling cascade is involved in the signal transduction process following TIGIT/DNAM-1-PVR engagement.
RESULTS: Blocking of DNAM-1 by an anti-DNAM-1 antibody and knockdown of DNAM-1 expression by lentiviral shRNA both resulted in deceased cytotoxicity of NK cells against alloantigen activated T cells, while over-expression of DNAM-1 via lentiviral transduction resulted in enhanced cytotoxicity. Blocking of TIGIT by an anti-TIGIT antibody and knockdown of TIGIT expression by lentiviral shRNA both resulted in increased cytotoxicity of NK cells against alloantigen activated T cells, while over-expression of TIGIT via lentiviral transduction resulted in decreased cytotoxicity. Increases in NK cytotoxicity against activated T cells through TIGIT knockdown could be overcome by blocking DNAM-1 signaling. Simultaneously, over-expression of DNAM-1 or knockdown of TIGIT expression resulted in an increase of the phosphorylation levels of Akt and ERK1/2 in NK cells after contacted with activated T cells, which could be overcome by pretreating NK cells with anti-DNAM-1 or PI3K small molecule inhibitor. Pretreating alloantigen activated T cells with anti-PVR also resulted in deceased cytotoxicity and Akt and ERK1/2 phosphorylation in DNAM-1 over-expression NK cells.
Conclusion: The paired receptor DNAM-1/TIGIT on the surface of NK cells compete the same PVR ligand on the surface of activated T cells and the DNAM-1/TIGIT axis is involved in the regulation of cytotoxicity of NK cells on alloantigen activated T cells through PI3K-Akt-ERK cascade phosphorylation. The DNAM-1/TIGIT expression balance may present as biomarkers for aGVHD and potential targets for aGVHD therapy.

Disclosures
No relevant conflicts of interest to declare.
IL-6 As a Potential Cell Released Marker of Prostate Cancer Cell Death
Authors: Gilazieva Zarema , Tezcan Gulcin , Garanina Ekaterina , Sakr Nawar , Arkhipova Svetlana , Vinogradov Lev , Rizvanov Albert , Khaiboullina Svetlana 
Published: October 2019

Abstract
Introduction: Interleukin 6 (IL-6) is proinflammatory cytokine which is produced by cell when Nod-like receptors (NLRs) are activated. Increased production of IL-6 was shown to promote tumor growth and metastasis.Therefore, it could be suggested that activation of NLRs could support malignancy. PC-3, prostate tumor derived cells, was shown to produce IL-6; however, little is known about the role of NLRs in regulation of cytokine secretion.
Method: PC-3 cells were seeded in 48 well plate cell culture plates and culturd in presence of VX765, Nigericin or Camptothesin. After 24 hours incubation, cell death was determined using Annexin V test. Additionally, IL-6 level was assessed in culture medium using Bio-Plex Pro™ Human Chemokine Panel (Biorad). Data was analyzed using One Way Anova, Post Hoc Tukey test, SPSS 20.
Results: Viability of PC3 cells was reduced when treated with Nigericin (37.74±0.33%) and Champtothesin (10.10±0.23%) as compared to untreated cells (P<0.01), while VX765 did not affect cell viability (4.78±0.28 %; P= 0.645). Nigerecin significatly increased IL-6 production in PC3 cells when compared to untreated cells. Interestingly, IL-6 levels in culture medium of PC3 cells treated with nigerecin was significantly higher than that in cells treated with VX765 and campthothesin (p< 0.001).
Conclusion: It appears that IL-6 can be produced by PC3 cells upon stmulation of NLRs. Since NLRs activation was associated with reduced cell vitality, we suggest that increased IL-6 level in culture medium could be the result of the cytokine release from dead cells.
This study was supported by RFBR Grant #18-34-01000 and Program of Competitive Growth of KFU.

Disclosures
No relevant conflicts of interest to declare.
Evaluation of Ocular Complications By Using Optical Coherence Tomography in Children with Sickle Cell Disease
Authors: Vatansever Esra , Vatansever Mustafa , Dinç Erdem , Temel Gulhan , Unal Selma 
Published: October 2019

Abstract
Purpose: This study was aimed to evaluate ocular abnormalities by using optical coherence tomography (OCT) in children with sickle cell disease (SCD) and to search the relationship between OCT and clinical findings.
Methods:The study included 108 eyes of 54 SCD patients as well as 110 eyes of 55 healthy subjects with no ocular pathology as a control group. A complete ophthalmologic examination was performed. Pupillary dilation was induced and macular sections were obtained with spectral domain OCT. Macular sections were evaluated according to the ETDRS map. Inner and outer retinal thicknesses were measured using the software in the device.
Results:Foveal splaying, temporal thinning of retinal thickness, and vascular tortuosity were found to be significantly higher in the patient group than the control group (p <0.0001 for all three parameters)(Figure-1).The foveal diameter (1592.39 ± 175.56 μ) was significantly increased in the patient group compared to the control group (1391.01 ± 175.56 μ) (p<0.0001) and the foveal depth was significantly decreased in the patient group (121.15 ± 26.83 μ) compared to the control group (146.1 ± 12.25 μ) (p<0.0001). The mean total retinal thickness was 253.53 ± 22.31 μ in the patient group and 261.03 ± 18.48 μ in the control group and the difference was statistically significant (p = 0.007). Likewise, central retinal thickness was significantly decreased in the patient group (219.35 ± 10.53 μ) compared to the control group (235.32 ± 12.51 μ) (p <0.0001).
Conclusions:Our study suggests that subclinical retinal involvement may occur in children with SCD. Foveal splaying, temporal thinning of retinal thickness, and vascular tortuosity are significant OCT findings in the children with SCD. It is seen that OCT is an important imaging method in addition to routine ophthalmologic examination to detecting ocular abnormalities in the early period and follow-up of this patient group.
Keywords :Sickle Cell Disease, Childhood, Optical Coherence Tomography
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
The Von Willebrand Factor Tyr2561 Allele Is a Gain-of-Function Variant and a Potential Risk Factor for Early Myocardial Infarction
Authors: Schneppenheim Reinhard , Hellermann Natalie , Brehm Maria Alexandra , Klemm Ulrike , Obser Tobias , Huck Volker , Schneider Stefan W , Denis Cecile V. , Tischer Alexander , Auton Matthew , Maerz Winfried , Xu Emma-Ruoqi , Wilmanns Matthias , Zotz Rainer B. 
Published: October 2019

Abstract
The frequent von Willebrand factor (VWF) variant p.Phe2561Tyr is located within the C4 domain, which also harbors the platelet GPIIb/IIIa-binding RGD sequence. To investigate its potential effect on hemostasis, we genotyped 865 patients with coronary artery disease (CAD), 915 with myocardial infarction (MI) and 417 controls (Ludwigshafen Risk and Cardiovascular Health Study (LURIC)) and performed functional studies of this variant. A univariate analysis of male and female carriers of the Tyr2561 allele ≤55 years revealed an elevated risk for repeated MI (Odds ratio: 2.53, 95% CI:1.07-5.98). The Odds ratio was even higher in females ≤55 years at a value of 5.93 (95% CI:1.12-31.24). Cone and plate aggregometry showed, that compared to Phe2561, Tyr2561 was associated with increased platelet aggregate size, both in probands' blood and with the recombinant variants. Inhibition of the VWF-GPIIb/IIIa interaction abrogated the effect. Microfluidic assays revealed that the critical shear rate for inducing aggregate formation was decreased to 50% by Tyr2561 compared to Phe2561. Differences in C-domain circular dichroism spectra resulting from Tyr2561 suggest an increased shear sensitivity of VWF, as a result of altered association of the C-domains that disrupts the normal dimer interface. In summary, our data emphasize the functional impact of the VWF C4-domain for VWF-mediated platelet aggregation in a GPIIb/IIIa-dependent manner and provide the first evidence that a functional variant of VWF plays a role in arterial thromboembolism.

Disclosures
Schneppenheim: SHIRE: Consultancy; CSL Behring: Consultancy.
Identification of a Novel CEBPE Enhancer Essential for Granulocytic Differentiation
Authors: Shyamsunder Pavithra , Shanmugasundaram Mahalakshmi , Mayakonda Anand , Teoh Weoi Woon , Han Lin , Lim Mei Chee , Fullwood Melissa , An Omer , Yang Henry , Jizhong Shi , Hossain Md.Zakhir , Madan Vikas , Koeffler H. Phillip 
Published: October 2019

Abstract
CEBPE is a member of the CCAAT/enhancer binding protein (C/EBP) family of transcription factors essential for granulocytic differentiation. CEBPE is expressed in a stage-specific manner during myeloid differentiation and regulates transition from the promyelocyte to the myelocyte stage. It is essential for secondary and tertiary granule formation in granulocytes. We and others found germline mutations of the CEBPE gene in patients with neutrophil-specific granule deficiency. Their neutrophils display atypical bilobed nuclei, lack expression of granule proteins and these patients often have frequent bacterial infections. Cebpe knock-out mice resemble this clinical phenotype displaying a block in terminal differentiation and absence of secondary granule proteins.
Given the tissue specific expression of CEBPE, we were interested in identifying genomic regions and factors that could regulate its lineage specific expression. Our CEBPE ChIP-seq in murine bone marrow cells showed binding of CEBPE to a region 6kb upstream of Cebpe gene. Chromosome conformation capture-on-chip (4C-seq) demonstrated an interaction between this putative regulatory element (6kb upstream region) and the core promoter of Cebpe. Analysis of available DNase-seq data sets revealed that the region bound by CEBPE displayed an open chromatin only in myeloid lineage cells. Further examination revealed binding of a myriad of hematopoietic transcription factors to the +6kb enhancer in HPC-7 (hematopoietic progenitor cells) and in 416B (myeloid progenitor cells), indicating that this region/enhancer might regulate the expression of CEBPE.
Targeting of this region using dCas9-KRAB in murine 32D cells caused significant downregulation of RNA and protein levels of CEBPE compared to control cells. These targeted cells also exhibited impaired granulocytic differentiation with lower transcript levels of secondary granule proteins (Ltf and Ngp).
To investigate further the role of the +6kb enhancer region in myelopoiesis, mice were generated with deletion of this region using CRISPR/Cas9 technology. Germ line deletion of the +6kb enhancer resulted in reduced levels of CEBPE and its target genes, accompanied by a severe block in granulocytic differentiation and a complete absence of CD11b+/Gr1hi population. This phenotype is nearly identical to our Cebpe KO mice.
In summary, we have identified a novel enhancer crucial for regulating Cebpe, and required for normal granulocytic differentiation.

Disclosures
No relevant conflicts of interest to declare.
Multidimensional Study on the Prognostic Factors of Acute Myeloid Leukemia Patients with FLT3-ITD mutations Based on the Next-Generation Sequencing Platform
Authors: Zhang Yi , Zhao Bide , Wang Yungui , Wang Dongmei , Pan Hanzhang , Wang Chengcheng , Lou Feng , Cao Shanbo , Jin Jie 
Published: October 2019

Abstract
Purpose: This research aimed to investigate the features and effects of FLT3-ITD mutations in Chinese acute myeloid leukemia (AML) population, and to explore whether ITD variant allele frequency (VAF), number of mutations, insertionposition and length will have influence on the patients' overall survival (OS) and relapse-free survival (RFS).
Methods:A total of 101 newly diagnosed AML (non-APL) patients with FLT3-ITD mutations from December 2013 to March 2018 were recruited. Their bone marrow or peripheral blood samples were sequenced by NGS 185-gene platform, then the sequencing data were analyzed in combination with the patients' clinical information.
Results:The most common concomitant mutations in FLT3-ITD patients were NPM1, DNMT3A, TET2, IDH2, RUNX1, and WT1. The patients' median VAF is 0.344 (0.084-0.487). Among 101 patients, 80 (79.2%) carried 1 ITD mutation, 17 (16.8%) carried 2, 3 (3.0%) carried 3, and only 1 (1.0%) carried 4. In most patients (76/101) the ITD fragments were inserted in the JM region. And the median length of the insertion fragments was 54 (12-252) bp. According to the NCCN guidelines 2018, the cutoff value of FLT3-ITDmutation VAF between different stages of prognosis is 0.33. However, in our research, it was better to set 0.30 (p=0.0465 for OS; p=0.4929 for RFS), but not 0.33 (p=0.4270 for OS; p=0.3828 for RFS), as the cutoff value to tell differences of survival. Meanwhile, the numbers of carried ITD was significantly related to prognosis. The patients with single ITD mutation were found to have better clinical outcomes than those with multiple ITD mutations (p= 0.0031 for OS; p=0.0001 for RFS). Patients with ITD insertion in the hinge region of JM had a trend of poorer OS. The insertion length of ITD fragment correlated with the patients' RFS, but not OS. The longer the insertion was, the worse RFS of the patient showed (p=0.0323, p trend=0.0049).
Conclusion:The VAF, the number of mutations, and the insertion length of the FLT3-ITDmutations will significantly affect the prognosis of AML patients in Chinese population.

Disclosures
Jin: College of Medicine, Zhejiang University: Employment; The National Natural Science Foundation of China: Research Funding.
The Role of High Molecular Weight Kininogen in Lymphoma Microvascular Density and Growth
Authors: Erdemir Huseyin Halil , Shukla Meenal , McCrae Keith R. 
Published: October 2019

Abstract
Background: Non-Hodgkin lymphoma (NHL) is a common malignancy in childhood and adolescence. The prognosis of relapsed and refractory NHL remains poor, and new approaches to therapy are needed. High molecular weight kininogen (HK) is a central component of the kallikrein-kinin system. Previous work has demonstrated that in vitro, the kallikrein-cleaved form of HK induces apoptosis of proliferating endothelial cells. Prior studies performed in our laboratory have demonstrated the increased angiogenesis and tumor growth in the absence of HK in B16F10 melanoma and Lewis lung carcinoma models. However, the role of endogenous kininogen in modulating the growth and of hematologic malignancy is unknown. To elaborate the role of HK in in the development and progression of a murine lymphomas, we assessed growth and vascularization of T-cell lymphoma xenografts in wild type (WT) and kininogen deficient (mKNG1-/-) mice.
Methods: Murine T-cell lymphoma (EL-4) cells (1 x 10^6) were implanted into wild-type and mKNG1-/- mice. Tumor size was measured using calipers and tumor volume was calculated using the formula V = length x width^2 x 0.52. Fifteen days after cell implantation, tumors were harvested, processed and analyzed by immunofluorescent staining with anti-CD31 antibodies to determine microvascular density (MVD).
Results: EL-4 lymphomas grew more rapidly and to larger sizes in mKNG1-/- mice compared to wild-type mice, with significant differences apparent by day 11 after tumor implantation (p<0.001). By Day 15, the volume of tumors in mKNG1-/- mice was approximately ~2-fold greater than in wild-type mice (Figure 1A). Weights of EL-4 Lymphomas were determined after 15 days of implantation and found significantly different between two groups (Figure 1B). To assess whether the difference in growth rates of lymphomas in wild-type and mKNG1-/- correlated with differences in angiogenesis, the microvascular density within tumors from these animals was compared by counting vessels stained with anti-CD31 antibodies (Figure 1D). Compared to wild-type mice, lymphomas in kininogen deficient mice demonstrated significantly greater microvascular density when either random fields were assessed (Figure 1C and 1D, P < 0.001).
Conclusion: These findings suggest that HK is an important endogenous regulator of angiogenesis and tumor growth in this T- cell Lymphoma model. Although the role of kininogen has been described in solid tumor models, this study is the first to demonstrate that kininogen specifically angiogenesis in a hematopoietic tumor. Further work is needed to understand the mechanisms underlying these findings and inform anti-angiogenic therapies to increase the options for patients with hematopoietic malignancies such as NHL.

Disclosures
No relevant conflicts of interest to declare.