bluuebunny/crossref_metadata_2025_split huggingface dataset:
FDC Like Cell Line HK With IL-2, IL-4 and OX40 Ligand Supports The Growth Of ATLL Cells
Authors: Kagami Yoshitoyo , Chihara Dai , Kato Harumi , Yoshida Noriaki , Kinoshita Tomohiro , Seto Masao 
Published: October 2019

Abstract
Adult T-cell leukemia/lymphoma (ATLL) is an aggressive neoplasm derived from CD4+ T-cells with HTLV-I infection, and its mechanisms of tumorigenesis still remain to be elucidated. The fact that tumor cells rarely proliferate in vitro is one of the most important problems to be solved. The establishment of cell line from ATLL patient samples has been difficult even in the presence of interleukins. Previously we established one cell line (HU-ATTAK) from acute or lymphoma types of 10 ATLL cases which did not proliferate in the presence of IL-2 and/or IL-4. HU-ATTAK is critically dependent on IL-2 and human umbilical cord vein endothelial cells (HUVEC) as feeder cells. In HU-ATTAK, adding anti-OX40 ligand antibody into the culture system completely inhibited its proliferation. So, OX40 ligand as well as L-2 and/or IL-4 is suggested be necessary for the proliferation of ATLL cells, and feeder cells may also confer the favorable environment. As the substitute of HUVEC, follicular dendritic cell like cell line HK which expresses OX40 ligand was used by introducing human OX40 ligand cDNA (HK-OX40L). When 9 ATLL patient samples were co-cultured with HK-OX40L in the presence of L-2 and/or IL-4, two ATLL cells proliferate vigorously only in the presence of both IL-2 and IL-4 simultaneously. These cell lines were confirmed to be derived from original tumor cells by array CGH analysis, and continued the growth for more than a year. Depletion of IL-2 and IL-4 made these cell lines stop growing within 6 days even on HK-OX40L. In the presence of IL-2 and IL-4, the conditions such as HK alone without OX40 ligand or OX40 ligand alone without HK made the cell lines growing for three months at most. In the presence of IL-2 and IL-4 without HK-OX40L, these cell lines vigorously proliferated for more than three months but finally stopped growing. These data suggested that for the growth of these two cell lines, the cell division is dependent on IL-2 and IL-4, and the maintenance of immortalization is dependent on OX40 ligand and HK cells. This culture analysis would provide important factors for cell growth of ATLL which will explore new targets for ATLL treatment.
*HK cells are kindly provided by Dr. Young S Choi at Ochsner Cancer Center, New Orleans.

Disclosures:
No relevant conflicts of interest to declare.
Unmasking Lymphoma Immune Reconstitution Inflammatory Syndrome Among HIV-Infected Individuals In The Center For AIDS Research Network Of Integrated Clinical Systems
Authors: Gopal Satish , Patel Monita , Yanik Elizabeth , Burkholder Greer , Reid Erin , Rodriguez Benigno , Deeks Steven , Mayer Kenneth , Moore Richard , Kitahata Mari , Eron Joseph , Richards Kristy L. 
Published: October 2019

Abstract

Introduction
Cohort studies have demonstrated increased incidence of Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) among HIV-infected individuals during the first 6 months after antiretroviral therapy (ART) initiation, perhaps due to unmasking immune reconstitution inflammatory syndrome (IRIS). Unmasking IRIS is characterized by the diagnosis of a new HIV-associated condition soon after ART that was not apparent prior to ART, coupled with evidence of ART effectiveness. It has been well described for opportunistic infections and Kaposi sarcoma. However, clinical characteristics and survival for unmasking lymphoma IRIS have not been described.

Methods
We studied lymphoma patients in the Centers for AIDS Research Network of Integrated Clinical Systems (CNICS) from 1996 until 2011. Unmasking lymphoma IRIS was defined as HL or NHL occurring within 6 months after ART initiation accompanied by a ≥0.5 log10copies/mL reduction in HIV RNA between values taken prior to ART and at lymphoma diagnosis. Differences in presentation and survival were examined between lymphoma IRIS and non-IRIS cases.

Results
Of 482 lymphoma patients, 48 (10%) met criteria for unmasking lymphoma IRIS. Of these, 10 (21%) had HL, 19 (40%) diffuse large B-cell lymphoma (DLBCL), 4 (8%) Burkitt lymphoma (BL), 9 (19%) primary central nervous system lymphoma (PCNSL), and 6 (12%) other NHL (Table). Median CD4 cell count at lymphoma diagnosis among IRIS cases was 163 cells/mL (interquartile range 67-302), and 54% had suppressed HIV RNA (< 400 copies/ml). No significant differences were identified between lymphoma IRIS and non-IRIS cases, with the exception of possible earlier stage (47% stage I/II versus 24%, p=0.03), more frequent hepatitis B/C co-infection (31% versus 19%, p=0.05), and more frequent prior AIDS illness (92% versus 79%, p=0.05), as well as expected lower HIV RNA at lymphoma diagnosis likely resulting from the IRIS case definition. Additionally, no differences in cumulative mortality 5 years after lymphoma diagnosis were identified between IRIS and non-IRIS cases, although there was a suggestion of increased early mortality among IRIS cases (Figure).

Conclusions
In a large HIV-associated lymphoma cohort in the United States, 10% of patients met a standardized unmasking lymphoma IRIS case definition. No major differences between lymphoma IRIS and non-IRIS cases were identified. Possible effects of subclinical lymphoma leading to care seeking behavior and subsequent ART initiation prior to lymphoma diagnosis could not be excluded.

Disclosures:
No relevant conflicts of interest to declare.
Cholesterol Levels In Patients With Newly Diagnosed Chronic Lymphocytic Leukemia: A retrospective Multicenter Study
Authors: Yavasoglu Irfan , Pektas Gokhan , Yilmaz Fergün , Akgün Gülsüm , Tombak Anil , Toka Bila , Dal Sinan , Ozbas Hasan , Cetin Guven , Donmez Ayhan , Bilgir Oktay , Tiftik Naci , Ertop Sehmuz , Ayyildiz Orhan M. , Sonmez Mehmet , Kadikoylu Gurhan , Tombuloglu Murat , Bolaman Zahit 
Published: October 2019

Abstract
Low cholesterol levels can be detected in solid tumors and hematological malignancies such as muliple myeloma. Moreover chloesterol levels reduced in some experimental studies of patients with chronic lymphocytic leukemia (CLL). In this retrospective multicenter study, lipid levels were retrospectively evaluated in 420 (264 male and 256 female with mean age 64 ± 11 years) patients with newly diagnosed CLL, according to the International CLL study group. 71 (28 male and 43 female with mean age 55 ± 9 years) healthy subjects as control group were included to this study. Lipid parameters such as total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), very low-density lipoprotein-cholesterol (VLDL-C), and triglyceride levels were measured with enzymatic/ calorimetric method and Architect C800 instrument. Low-density lipoprotein-cholesterol (LDL-C) levels were calculated according to Friedwald formula. Lipid parameters between two groups were compared with Mann-Whitney U test. A value of p< 0.05 was accepted as statistically significant. According to Binet classification, 60% of patients were in stage A, while 25% of them were in stage C. In CLL patients, the levels of TC, HDL-C, and LDL-C were lower than those of control group (p=0.001). There was no significantly difference for triglyceride and VLDL-C levels between two groups (p>0.05) (Table-1). The levels of TC, LDL-C, and HDL-C in the patients with stage C were lower than those of both stage A and stage B (Table-2). Low cholesterol levels in patients with CLL may occur due to increased use of cholesterol by lymphocytes.Table-1Lipid parameters in control group and CLL patientsCLL (N:420)Control (N:71)P valueTC (mg/dl)175±41217±36<0.001HDL-C (mg/dl)37±1153±14<0.001LDL-C (mg/dl)108±30131±29<0.001Triglyceride (mg/dl)140±71147±68>0.05VLDL-C (mg/dl)31±1731±17>0.05Table 2Lipid parameters in the patients according to Binet classificationStage A (n:255)Stage B(n:61)Stage C(n:104)P valueTC(mg/dl)183±38179±43156±40<0.001HDL-C(mg/dl)40±1237±1232±12<0.001LDL-C (mg/dl)112±28110±3398±30<0.001Triglyceride (mg/dl)141±76136±53141±68>0.05VLDL-C(mg/dl)30±1535±3331±13>0.05

Disclosures:
Sonmez: Novartis Pharmaceuticals Corporation, Turkey: Membership on an entity’s Board of Directors or advisory committees.
The Study On The Relationship Between c-Myc and Hif-1 α In u266 Cell Line
Authors: Ma Yanping 
Published: October 2019

Objective
The aim of study is to explore the relationship of C-MYC and HIF-1 α in multiple myeloma cell lines U266,and research the influence of C-MYC/HIF-1 α on VEGF, in order to find a new target for clinical treatment MM.

Methods
Transfect the siRNA c-myc plasimd,siRNA HIF-1 α plasimd and mock plasimd into U266 cell lines respectively.RT-PCR was used to detected the expression of C-MYC and HIF-1αin different group, (fetch the CT value, using 2-ΔΔCT to calculate) .The expression of protein of C-MYC.HIF-1αand VEGF was measured with Western-blot in different group,((using the ratio of purpose gene and internal reference β-actin expression as the final results).

Results
compared with the control group, the expression of C-MYC, HIF-1 α and VEGF was decreased in the siRNA c-myc group, which differences are statistically significant (p<0.05), the expression of HIF-1 α and VEGF was decreased in the siRNA c-myc group ,which differences are statistically significant (p<0.05),the expression of C-MYC have no obvious decline compared with control group.

Conclusion
The expression of HIF-1 α depends on the regulation of C-MYC gene, synthesis and secretion of VEGF depends on C-MYC/HIF-1 α, C-MYC/HIF-1 α may serve as a new target for treatment of MM.

Disclosures:
No relevant conflicts of interest to declare.
Expression Of Cancer-Testis Antigen In Multiple Myeloma
Authors: Ji Jingna , Liu Shangqin , He Li , Xu Chaoping , Hu Guiying , Zhang Yicheng , Ma Zi , Huang Jianying 
Published: October 2019

Abstract
In order to detect the CTAs expression in the cell lines and in patients with multiple myeloma (MM). Reverse transcriptase polymerase chain reaction (RT-PCR) detects the mRNA expression of MAGE-C1/CT7, SSX1, SSX2 and SSX4 in MM cell lines RPMI8226 and U266 and patients. Collect clinical MM patients with bone marrow specimens of 25 cases,18 cases of healthy volunteers as a control, The expression frequency in MM patients of CTA gene is parametric statistical analysis with age,gender,the amount of plasma cells,clinical stage and MM type. The CTA members we detected all express in RPMI8226 and U266 cell lines, the expression frequencies of MAGE-C1/CT7 of SSX1, SSX2 and SSX4 in 25 cases bone marrow of MM patients are as follows: 28%（7/25）、80%（20/25）、40%（10/25）、68%（17/25）. 18 cases of healthy volunteers with bone marrow does not express the gene. Four kinds of genes in the bone marrow of patients with simultaneous expression of two or more frequency 80%（20/25）, at least have an expression of the frequency 88%（22/25）. Expression of SSX1 and SSX4 in different disease stage was statistically significant(P &lt;0.05), expression frequency was mainly for patients in the phase Ⅲhigher thanⅠand Ⅱ. Expression of MAGE-C1/CT7 and SSX2 in the period of disease was not statistically significant(P﹥0.05), Detected by the four CTA with age, gender, MM type and the volume of plasma cells was no significant difference(P﹥0.05). It was suggested MAGE-C1/CT7, SSX1, SSX2 and SSX4 gene in MM cell lines RPMI8226 and U266 and MM patients can co-express, while do not express in healthy people. In the expression frequency, SSX1 and SSX4 relate with MM clinical stage,MAGE-C1/CT7 and SSX2 do not relate with clinical stage,the 4 CTAs do not relate with gender,age,clinical type and volume of plasma cells. it provides theoretical support for the CTA vaccine in multiple myeloma immunotherapy.

Disclosures:
No relevant conflicts of interest to declare.
Bortezomib-Based Chemotherapy Regimens Can Inmprove The Complete Remission In Newly Diagnosed Multiple Myeloma Patients With Bcl-2 and Survivin Overexpression
Authors: Wen Zeng , Lifang Huang , Zhang Yicheng , Hanying Sun , Wenli Liu , Fankai Meng , Jianfeng Zhou 
Published: October 2019

Abstract
Compared with the traditional chemotherapeutic agents, Bortezomib-based chemotherapy regimens can increase the complete remission and improve the prognosis significantly in the newly diagnosed multiple myeloma patients. Molecular markers which can predict the chemotherapeutic efficacy and safety could help the physicians to make the right decisions and benefit the patients, and it has been one of most interesting research fields in MM. It has been reported that Survivin and Bcl-2 was involved in the adverse clinical events and therapeutic resistance, respectively. In order to investigate the relationship between the Survivin and Bcl-2 expression and the different regimens therapeutic efficacy, we retrospectively studied the proteins expression in the bone marrow biopsy specimens by inmmunohistochemistry and the efficacy of different chemotherapy regimens in the newly diagnosed MM in a single center of Tongji Hospital. Total 59 newly diagnosed MM were admitted into this study. The positive expression rate for Survivin and Bcl-2 was 35.3%（n=21）and 73.5%（n=43）, respectively. The protein expression had no relationship with the Durie-Salmon and International Staging System stratification, which suggested that Survivin and Bcl-2 were not responsible for the clinical manifestations. Bortezomib-based regimens (n=22) could effectively decrease the tumor burden and achieve response (CR+PR: 67.5% ). The non-bortezomib regimens (n=37), containing VAD(T), MP(T), TAD, were effective in the absent of Survivn and Bcl-2 expression(n=7; CR+PR 62.5% ). When Survivin and Bcl-2 were single or double positive (n=30), the newly diagnosed MM patients had no response for non-bortezomib regimens with none reaching complete remission or partial remission (p=0.0088). According to this study, we recommend the newly diagnosed MM which were Survivin and Bcl-2 single or double positive by inmmunohistochemistry received the regimens containing bortezomib for anti-myeloma therapy.

Disclosures:
No relevant conflicts of interest to declare.
Genomic Characterization Of Newly Established Two Distinct Patient-Driven Multiple Myeloma Cell Lines (SNU_MM1393_BM and SNU_MM1393_SC) From a Single Patient
Authors: Koh Youngil , Ahn Kwang-Sung , Lee Chansu , Jung Woo June , Lee Hyun Jung , Lee Dong Soon , Kim Hyo Jung , Yoon Hwi-Joong , Yoon Sung-Soo 
Published: October 2019

Abstract
To establish patient-derived multiple myeloma (MM) cell lines, mononuclear cells obtained from a MM patient’s bone marrow were directly injected via tail vein into a NRG/SCID mouse. Fourteen weeks after injection, tumor developed at subcutis and bone marrow (BM) in the same mouse. We separated and cultured cells from these two sites (subcutis and BM) and established two cell lines originating from a single patient (SNU_MM1393_BM and SNU_MM1393_SC). In cytogenetic analysis, karyotype of newly established two MM cell lines showed tetraploidy which is different from the karyotype of the patient (diploidy) indicating clonal evolution. In FACS analysis, the expression of CD138 and CD45 was detected in both cell lines. Response to IL-6 and soluble IL-6 receptor was different between the two cell lines. Moreover, SNU_MM1393_SC showed higher degree of resistance against proteasome inhibitor (bortezomib) compared to SNU_MM1393_BM. However, two cell lines were both sensitive to histone deacetylase inhibitor (panobinostat).
When whole exome sequencing was performed using the DNA of these two cell lines, a set of somatic mutations involving Wnt signaling and NF-kB pathway were detected in both cell lines. Additional somatic mutations of JAK1, PLCG1, IRS2, and HGS which are known to interact with JAK/STAT pathway were detected in SNU_MM1393_BM. Whereas, additional somatic mutations of EGFR, HSP90AB1, CFDR, and CALML5, which are known to interact with growth factor cell signaling pathway were detected in SNU_MM1393_SC.
These findings highlight the importance of interactions between tumor and tumor microenvironment as the myeloma progresses and will pave a way to more effective selection of targeted agents according to specific tumor characteristics obtained in the process of disease progression.

Disclosures:
No relevant conflicts of interest to declare.
Aptamer TY04 Inhibits Multiple Myeloma Cell Growth Via Cell Cycle Arrest
Authors: Liu Jing , Dai Hong-Juan , Ma Bian-Ying , Song Jian-Hui , Chen Hui-yong , Zhou Jia-Jie , Tang Shi-Jun , Yao Shan , Ye Mao , Tan Wei-Hong 
Published: October 2019

Abstract
Multiple myeloma (MM), also known as plasma cell myeloma, is characterized by accumulation of clonal plasma cells in the bone marrow and overproduction of monoclonal immunoglobulin (Ig) in the blood or urine. MM accounts for approximately 10% of all hematologic malignancies. Despite recent advances in the understanding and treatment of this disease, MM remains an incurable disease in the vast majority. With conventional chemotherapy, the 5-year median survival rate for MM patients is approximately 25%. Aptamers are single-stranded RNA or DNA sequences that bind to target molecules with high affinity and specificity. Compared with antibodies, aptamers have unique advantages including easy chemical synthesis and modification, low toxicity, lack of immunogenicity, and rapid tissue penetration, Based on these advantages, aptamers show great potential for therapeutic application. The aptamer TY04 is a single-stranded DNA (ssDNA) generated by a method named cell-based systematic evolution of ligands by exponential enrichment (cell-SELEX), We found TY04 strongly inhibited the growth of multiple myeloma cell lines including MM1.S, NCI-H929, KM3 and OPM2,The concentration of TY04 to inhibit 50% cell growth (IC50) on MM1.S was 3.89 μM. In contrast, TY04 had no effect on the growth of non-tumor cell lines — immortal B lymphoblastoid cell lines. Next, we used MM1.S cell line as the model to study the mechanism of TY04 anti- multiple myeloma. Flow cytometry analysis showed that TY04 with the sequence specifically bind to MM1.S cells when compared with unselected ssDNA library control. To investigate whether the target molecules of TY04 are membrane proteins on cell surface, MM1.S cells were treated with trypsin and proteinase k for 2 or 10 minutes before incubation with TY04. The result revealed that TY04 lost partly recognition ability on treated cells, indicating that the target molecules were most likely membrane proteins. Furthermore, we evaluated the cell cycle distribution of MM1.S after TY04 treatment. We found that TY04 significantly caused cell-cycle arrest in G2/M phase. The percentage of G2/M phase cells increased from 30.1±1.56 to 53.2±6.36. To identify the underlying molecular mechanism, G2/M-related proteins were assayed by flow cytometry. Following TY04 treatment, a concomitant inhibition of ERK1/2, cyclin B, CDK1 and γ-tubulin expression occurred. Meanwhile, human cell cycle PCR array was used to analyze the expression of 84 genes key to cell cycle regulation in TY04-treated MM1.S cells. Our results indicated that aptamer TY04 decreased the genes expression of CCNB1, CCNB2, BIRC5, BRCA1 and CCNH, which were involved in the progress of G2/M phase. All these results are significant in that they provide a framework for further exploring the use of TY04 as a novel anti-multiple myeloma agent.

Disclosures:
No relevant conflicts of interest to declare.
CD28 Regulates Metabolism In Multiple Myeloma For Cell Survival and Proliferation
Authors: Utley Adam T , Murray Megan , Carlson Louise M , Farren Matthew R. , Nair Jayakumar R , Lee, MD Kelvin P 
Published: October 2019

Abstract
Multiple myeloma is an incurable hematological malignancy of transformed plasma cells. Many cellular interactions and soluble factors have been demonstrated to play a role in myeloma pathogenesis; however, novel targets to enhance therapeutic intervention are needed. We have demonstrated that CD28 signaling in myeloma cells supports their survival during chemotherapeutic challenge in vitro and in vivo. However, the cellular mechanisms by which CD28 confers this survival advantage to myeloma cells are not completely understood.
CD28 is best characterized as the canonical T cell co-stimulatory molecule. During T cell activation, CD28 signaling induces glycolysis, a metabolic program required for T cell proliferation and functional maturation. In the absence of glycolysis, T cells utilize fatty acid oxidation for energy production through the mitochondria. However, the way in which CD28 regulates metabolism in multiple myeloma is not well understood. Here we present evidence that CD28 signaling induces glut1 expression, and that poisoning the glycolytic pathway inhibits proliferation and survival of myeloma cells.
AMPK, an energy sensitive kinase known to regulate metabolism by driving fatty acid oxidation, is normally activated when cellular energy levels are low. Interestingly, poisoning glycolysis with a glucose analogue that cannot be processed (2DG) leads to AMPK inhibition in myeloma cells. Furthermore, pharmacological activation of AMPK by AICAR, an AMP analogue, is not sufficient to rescue myeloma cell proliferation from glycolytic inhibition and in fact increases cell death (p<.01 from no treatment, p<.05 from 2DG). This evidence suggests that multiple myeloma cells are absolutely dependent upon CD28-mediated glycolysis for proliferation and survival, and that myeloma cells cannot utilize fatty acid oxidation as a subsidiary metabolic pathway for proliferation in the absence of glycolysis. This understanding will allow us to target metabolism in multiple myeloma as a novel therapeutic strategy through pharmacological targeting of the CD28 pathway. This approach can be quickly translated into the clinic, as there are FDA approved drugs which activate AMPK (Metformin) and block CD28 signaling (Abatacept).

Disclosures:
Off Label Use: Abatacept, purpose to prevent CD28-mediated cell survival in multiple myeloma Metformin, purpose to activate AMPK in driving multiple myeloma cell death.