bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hedgehog Pathway: A New Target for B-Cell Lymphomas an Multiple Myeloma.
Authors: Dierks Christine , Grbic Jovana , Zirlik Katja , Mertelsman Roland H. , Warmuth Markus 
Published: October 2019

Abstract
The hedgehog pathway (Hh) is one of several key developmental pathways whose deregulation is known to induce tumor formations in mice and humans including medulloblastoma or basal cell carcinoma. Certain human solid tumors (prostate cancer, pancreatic cancer) have been identified which require sustained Hh-Gli signaling for proliferation.
Here, we show for the first time a crucial role of Hedgehog signalling in hematopoietic malignancies. In our experiments, we demonstrate that the Hh pathway is essential for survival of more than 70% of primary lymphomas extracted from transgenic Eμ-Myc mice or Cdkn2a−/− mice (INK4Arf−/−). 34 Myc-lymphomas (10 B-cell lymphomas, 12 plasmablastomas, 10 plasmocytomas, 2 mixed lymphomas) and 10 Cdkn2a−/− lymphomas were isolated from bone marrow, spleen or lymph nodes from diseased mice and expanded on bone marrow stroma cells from Cdkn2a−/− mice. Inhibition of Hh signaling by Cyclopamine (2–5 μM), an alkaloid which inhibits smoothened activation, induced apoptosis in 80% of B-cell lymphomas, 82% of plasmablastomas and 60% of multiple myelomas without affecting stroma proliferation. Protein or transcript levels of Hh pathway downstream targets, such as Gli1, Ptch or Bcl2 but not BMI1 were downregulated after 24h treatment with Cyclopamine. Overexpression of Bcl2 or Gli3 could inhibit apoptosis induction by Cyclopamine.
High levels of Ihh protein could be detected in our stroma cell line as well as in bone marrow stroma from various mouse strains. Stimulation of lymphoma cells with recombinant Shh or Ihh protein could overcome apoptosis induction induced by removal of the stromal layer, indicating Ihh as a mediator between stroma and lymphoma cells.
Injection of luciferazed lymphoma cells into C57Bl6 mice induced lymphoma formation within 2–6 weeks, as shown by Xenogen luciferase imaging. Subcutaneous treatment of lymphoma injected mice with Cyclopamine could inhibit lymphoma formation and even reduce lymphoma mass in mice with already fully developed lymphomas, indicating stroma-lymphoma interaction through hedgehog signaling as an important survival mechanism for lymphoma cells also in vivo.
Our data demonstrates that proliferation and survival of a majority of B-cell lymphomas and plasmacytomas is dependent on intact Hh signaling in vitro and in vivo, suggesting disruption of this pathway as a novel approach in lymphoma therapy.
Dendritic Cell-Induced T Cell Non-Responsiveness Is Mediated by FOXP3+ and TGF-beta+IL-10+ CD4+ T Cells.
Authors: Berneman Zwi N. , Cools Nathalie , Van Tendeloo Viggo F.I. , Lenjou Marc , Nijs Griet , Van Bockstaele Dirk R. , Ponsaerts Peter 
Published: October 2019

Abstract
Dendritic cells (DC), the professional antigen presenting cells of the immune system, exert important functions both in induction of T cell immunity as well as of tolerance. Previously, it was accepted that the main function of immature DC (iDC) in their in vivo steady state condition is to maintain peripheral tolerance to self-antigens and that these iDC mature upon encounter of so-called danger signals and subsequently promote T cell immunity. However, a growing body of experimental evidence now indicates that traditional DC maturation can no longer be used to distinguish between tolerogenic and immunogenic properties of DC. In this study, we compared the in vitro stimulatory capacity of immature DC (iDC), cytokine cocktail-matured DC (CC-mDC) and poly I:C-matured DC (pIC-mDC) in the absence and presence of antigen. All investigated DC types could induce at least 2 subsets of regulatory T cells. We observed a significant increase in both the number of functionally suppressive transforming growth factor (TGF)-beta+ interleukin (IL)-10+ T cells as well as of CD4+CD25+FOXP3+ T cells within DC/T cell co-cultures as compared to T cell cultures without DC. The induction of these regulatory T cells correlates with in vitro T cell non-responsiveness after co-culture with iDC and CC-mDC, while stimulation with pIC-mDC resulted in reproducible cytomegalovirus pp65 or influenza M1 matrix peptide-specific T cell activation as compared to control cultures in the absence of DC. In addition, the T cell non-responsiveness after stimulation with iDC was shown to be mediated by TGF-beta and IL-10. Moreover, the suppressive capacity of CD4+ T cells activated by iDC and CC-mDC was shown to be transferable when these CD4+ T cells were added to an established T cell response. In contrast, addition of CD4+ T cells stimulated by pIC-mDC made responder T cells refractory to their suppressive activity. In conclusion, we hypothesize that DC have a complementary role in inducing both regulatory T cells and effector T cells, where the final result of antigen-specific T cell activation will depend on the activation state of the DC. This emphasizes the need for proper DC activation when T cell immunity is the desired effect, especially when used in clinical trials.
Identification of Human Cd8+ and Cd4+ Lymphocytes with Unique Drug Resistance Activity by Imaging the Drug Transport Kinetics at the Single Cell Resolution.
Authors: Shirihai Orian , Biran Israel , Bertelsen Kirk M. , Greenblatt David J. , Deutsch Motti , Zurgil Naomi 
Published: October 2019

Abstract
Drug resistance activity of transporters such as P-glycoprotein within lymphocytes may have significant consequences to the pharmacologic treatment of HIV, immune suppression, autoimmune diseases, and leukemia, as many drugs are substrates of the MDR transporters. Variability in MDR activity may be a significant barrier in the evaluation of the predicted effect of a drug and on the therapeutic dosage necessary to overcome MDR activity. Current methodologies to measure MDR, e.g. FACS, do not provide kinetic measurements of dye efflux at the resolution of the individual cell thereby missing the existence of functionally different subpopulations. To overcome these limitations, we developed a new imaging device that is comprised of thousands of transparent pico-wells (20 micron in diameter) organized in a honeycomb structure that enables real-time visualization and sequential quantification of drug transport rate from thousands of individual living lymphocytes, thereby generating MDR kinetic data. Fluorescence microscopy was used to measure DiOC2 dye (a common fluorescent probe for P-glycoprotein transport) retention from individual human CD4+ or CD8+ lymphocytes over time. Kinetic analysis of dye export from CD4+ and CD8+ cells revealed the occurrence of subpopulations of cells with unique sensitivity to MDR inhibitors. When the MDR inhibitor Verapamil was used, CD8+ lymphocytes demonstrated a greater magnitude of inhibition compared to CD4+ lymphocytes. Remarkably, CD4+ lymphocytes contained a sizable subpopulation which was verapamil insensitive, suggesting the existence of a non-P-glycoprotein drug export activity in these cells. These findings indicate that functional MDR assays, which are based on transport kinetics, can efficiently dissect the heterogeneous lymphocyte cell populations and identify unique subsets of cells with increased activity.
Figure Figure
Cyclosporine Toxicity and Vascular Endothelial Growth Factor (VEGF).
Authors: Kubisz Peter , Grandtnerova Barbara , Laca Ludovit , Stasko Jan 
Published: October 2019

Abstract
Vascular endothelial growth factor (VEGF) is a potent angiogenic and endothelial cell-specific mitogen. Expression of VEGF mRNA is increased under hypoxia/ischemia conditions. Its role in the setting of clinical kidney transplantation (Tx) is currently under investigation. Majority of works are dealing with gene polymorphisms and/or renal tissue expression. The aim of the study was to evaluate serum VEGF concentrations after kidney Tx and its relationship to graft outcome. Thirty-five adult patients (17 male, 18 female) treated with cyclosporine A or tacrolimus, at least 1 month after Tx and 22 healthy control (17 male, 5 female). Patients with acute rejection or infection were excluded. Serum VEGF (S-VEGF) concentrations were measured by Quantikine Immunoassay, RD Systems. S-VEGF concentrations were significantly higher after Tx than in healthy control (556 ± 463 pg/ml, vs 145 ± 74 pg/ml, p< 0.0001). No correlation was found with age, gender, time after Tx or type of calcineurin inhibitor. However, a significant correlation was found between basaline S-VEGF and S-creatinine (p< 0.05), S-VEGF and S-creatinine six months after Tx (p< 0.01) and between S-VEGF and cyclosporine toxicity defined as a gingival hyperplasia or biopsy proven nephrotoxicity (202 ± 121 pg/ml in stable patients vs 741 ± 436 pg/ml in cyclosporine toxicity group, p< 0.001). During six months of follow up, the kidney graft function was stable in 100% of patients with S-VEGF concentration ≤ 145 pg/ml but only in 50% of patients with S-VEGF concentration > 145 pg/ml (p < 0,05). We conclude that the S-VEGF concentration seems to be a marker of cyclosporine toxicity and prospective graft function deterioration. S-VEGF could be helpful to select patients who would have benefit from an early switch of immunosuppression.
The MYST Domain Histone Acetyltransferase TIP60 Interacts with and Is as a Coactivator of the Myeloid Transcription Factor C/EBPα.
Authors: Bararia Deepak , Trivedi Arun K. , Zada Abdul Peer , Behre Hermann M. , Behre Gerhard , Bohlander Stefan K. 
Published: October 2019

Abstract
Recently, we and others have shown that protein-protein interactions play an important role in the pathogenesis of leukemia, and that the transcription factor C/EBPα is a key player in granulopoiesis and leukemogenesis. In the present study, we sought to identify C/EBPα interacting proteins. A glutathione-S-transferase-C/EBPα fusion protein was used to pull down interacting proteins from U937 nuclear extracts. These proteins were analyzed by 2-D gel electrophoresis, 1-D nano LC and identified by MALDI or MALDI-TOF/TOF. Several novel C/EBPα interactors were identified including known proteins like Rb, hnRNP and E2F4. Two novel interactions, one of cell cycle regulator protein MCM5 and the other with the MYST domain histone acetyl transferase TIP60 with C/EBPα were further confirmed by using pull-down and co-immunoprecipitation experiments. TP60 was able to markedly enhance C/EBPα mediated transcription in reporter gene assays, suggesting that TIP60 is a co-activator of C/EBPα. This co-activator function of TIP60 was dependent on an intact histone acetyl transferase domain and on the C/EBPα DNA binding domain. TIP60 was found to be associated with the human C/EBPα promoter in-vivo in a chromatin immunoprecipitation assay with a concomitant increase in histone H3 and H4 acetylation. Furthermore, we observed a lower expression of TIP60 mRNA in U937 CD11b− compared to retinoic acid induced U937 CD11b+ cells suggesting that higher TIP60 expression is associated with myeloid differentiation. There was also a marked correlation between the expression levels of TIP60 and C/EBPa in normal bone marrow, chronic myeloid leukemia and acute myeloid leukemia samples with t(8;21), inv(16) and t(15;17). This finding further confirms the functional synergism between C/EBPα and TIP60 and suggests that TIP60 might be an important player in leukemogenesis.
Inhibition of the TGFβ Pathway in Primary Effusion Lymphoma.
Authors: Di Bartolo Daniel , Cesarman Ethel 
Published: October 2019

Abstract
Kaposi’s sarcoma-associated herpesvirus (KSHV) interferes with a number of cellular pathways involved in apoptosis and cell cycle regulation. Since TGFβ is a potent inducer of cell cycle arrest and apoptosis in hematopoietic lineages, we were interested in examining whether TGFβ signaling is blocked in primary effusion lymphoma (PEL) and if so whether KSHV plays a role in this repression. Treatment with TGFβ1 had no effect on either cell cycle or apoptosis in several PEL cell lines. The defect in this pathway was found to be a lack of the TGFβ type II receptor (TβRII), expression. Upon transfection of TβRII, PEL cell lines become responsive to TGFβ1 treatment. We are currently examining how TβRII transcription is regulated in PEL. The lack of TβRII expression in some tumors has been attributed to altered chromatin structure due to hypoacetylation. Treatment with both a demethylating agent and a histone deacetylase inhibitor resulted in expression of this receptor in PEL. Several KSHV latent gene products are known to interact with or recruit p300, a histone acetyltransferase, for other cellular processes. It is possible that the interaction of one or more of these viral gene products with p300 may serve to prevent it from acetylating histones associated with the TβRII gene and thereby repress its transcription. We have also found by ELISA that PEL cells secrete a higher level of TGFβ1 than other TGF β responsive B cell lines. Increased secretion of TGFβ1 in conjunction with downregulation of TβRII may be a mechanism by which KSHV creates an immunosuppressed environment while allowing infected cells to grow free from the constraints of this pathway.
MKC-1 Is a Novel Agent That Induces Cell Cycle Arrest and Disrupts Multiple Survival Pathways in Hematologic Cancers.
Authors: Denny Trisha A. , Chen Xiaoru , Waters Cassandra L. , Burke Patricia A. , Fletcher Graham C. , Xu John , Sidor Carolyn F. , Minden Mark D. , LaVallee Theresa M. , Bray Mark R. 
Published: October 2019

Abstract
MKC-1 is a novel, orally active cell cycle inhibitor with in vitro and in vivo activity against a wide range of human solid tumor cell lines, including multi-drug resistant cell lines. MKC-1 has been tested in over 270 patients to date and is currently in Phase II clinical trials. The strong pre-clinical activity of MKC-1 towards solid tumor lines and signs of efficacy in the initial clinical evaluation with lack of neuropathy and cardiotoxicity suggests that MKC-1 may also be of clinical benefit in the treatment of hematopoietic cancers. The antiproliferative activity of MKC-1 was examined against a panel of hematopoietic cell lines including HL-60, U937, MV4;11, THP-1, Jurkat, and OCI-AML 1–5. MKC-1 showed potent and dose-dependent activity towards these cell lines, with IC50 values in the range of 20 – 400 nM. MKC-1 also inhibited in vitro growth of primary cells derived from AML and CML patients. Additionally, MKC-1 showed enhanced activity with Ara-C in combination studies in vitro when added either simultaneously or sequentially using the cell line OCI-AML 4. Binding studies have shown that MKC-1 binds to the colchicine binding site of tubulin and to members of the importin beta family of proteins. Consistent with these results, cell cycle arrest in the G2/M phase of the cell cycle followed by apoptosis was observed in cell lines and patient samples treated with MKC-1. Immunofluorescence analysis of cells treated with MKC-1 revealed that the drug induced a disruption of the microtubule network and the formation of aberrant mitotic spindles. Furthermore, MKC-1 was also shown to induce a dose-dependent reduction in the levels of both phospho-Akt and phospho-p70S6K kinases through Western blot analysis of treated THP-1 cells. In conclusion, our results indicate MKC-1 arrests the cell cycle and disrupts multiple survival pathways to induce apoptosis in hematopoietic cell lines and patient samples. These results suggest that MKC-1 may have clinical potential in the treatment of leukemia either alone or in combination with other agents. Phase I trials in hematological cancers are currently being explored.
Re-Evaluation of Prognostic Value of Seric LDH and β2-Microglobulin Levels among 248 CLL with a Median Follow-Up Time More Than 6 Years.
Authors: Desablens Bernard , Garidi Réda , Gruson Bérengère , Vaida Iona , Brevet Marie 
Published: October 2019

Abstract
Except in MD Anderson Center, seric β2-m is not recognized as a strong prognostic factor in CLL and seric LDH too, despite this factor is more and more important among lymphoma with a continuous prognostic value. So we studied 248 CLL aged less than 75 yrs, without elevated creatininemia or evident hemolysis. There were 201 pts with stage A, 29 B and 18 C and sex-ratio M/F was 1.48. Median survival time is 10 yrs and median “corrected” (i.e., without not CLL-related deaths including those due to a solid tumor because this complication is independent of initial prognostic factors of CLL) survival time is 10.9 yrs. LDH values range from 0.55 to 3.54 N, and mean and median values are 1.05±0.40 and 0.96 N. LDH level is only correlated with blood lymphocytosis and hemoglobin levels studied as continuous values (p=0.015 and 0.002). When looking at “corrected” survival, all cut-off values from 0.75 N, N, 1.25 N… to 2.5 N are statistically significant but the best one is 1.25 N (p=6 10−6). β2-m values range from 0.81 to 16.5 mg per l, and mean and median values are 2.67±1.75 and 2.15 mg. When studied as continuous values, β2-m is linked to age, number of lymphoid areas according to Binet’s system, and hemoglobin and platelets levels (p=0.0009, <10−4, <10−4 and 0.003) but age was ruled out among patients less than 70 yrs. For “corrected” survival, all cut-off values except 1.25 mg were significant and for example p value at 4 mg level as usually used by MD Anderson Center, is 5 10−5. However we find that the best cut-off value is 3 mg (p<10−6) and this value is statistically independent from LDH one in Cox model. We tried to combine these 2 cut-off values. Product of observed LDH/1.25 N × observed β2-m/3 mg were pertinent to isolate pts with a very poor prognosis (median “corrected” survival time about 6 yrs) but size of pejorative subgroups were small (≈25% of all pts) and other subgroups were not statistically significant. When adding ratios, results were similar. So we find that the best system is to count the number of pejoratives factors, i.e., LDH > 1.25 N and β2-m > 3 mg. Global median survival times of the 3 defined groups were respectively > 10.8 yrs (160 pts), 6.5 yrs (68 pts) and 4.2 yrs (20 pts) with a p value < 10−6. This system is also efficient among stage A pts (p=10−5). When we compare this prognostic system with Binet’s and Rai’s prognostic systems, Cox model rules out Binet’s one and keeps Rai’s system and ours (p=0.01 and 0.0005). Among patients with stage A, Cox model keeps only this LDH and β2-m system. We conclude on the strong prognostic values of seric LDH and β2-m initial levels in CLL and we claim that these 2 simple biological parameters have to been compared with “modern” biological prognostic factors of CLL.
Phase I Study of AVE9633, an AntiCD33-Maytansinoid Immunoconjugate, Administered as an Intravenous Infusion in Patients with Refractory/Relapsed CD33-Positive Acute Myeloid Leukemia (AML).
Authors: Giles Francis , Morariu-Zamfir Rodica , Lambert John , Verstovsek Srdan , Thomas Deborah , Ravandi Farhad , Deangelo Dan 
Published: October 2019

Abstract
AVE9633 is an immunoconjugate created by conjugation of the cytotoxic maytansinoid, DM4, to the monoclonal IgG1 antibody, huMy9-6 (average of 3.5 molecules of DM4 per antibody). The huMy9-6 antibody is a humanized version of a murine monoclonal antibody, My9-6, which is specific for the CD33 antigen expressed on the surface of myeloid cells, including the majority of cases of AML. Because CD33 has little expression outside the hematopoietic system, it represents an attractive target for antibody-based therapy in patients with AML. The humanized antibody, huMy9-6, binds to the CD33 antigen with an apparent KD in the range of 10−10 M. Maytansinoids are anti-mitotics that inhibit tubulin polymerization and microtubule assembly, inhibiting cells during the G2/M phase of the mitotic cycle. In order to link maytansinoids to antibodies via disulfide bonds, a new thiol-containing maytansinoid (DM4) was synthesized. Attachment of potent maytansinoids to an antibody via disulfide bonds provides a satisfactory stability in the bloodstream. After the conjugate is bound at the specific tumor site it is internalized and the cytotoxic agent is released within the target cell. A phase I study of AVE9633 is being conducted in patients with refractory/relapsed CD33+ AML. The study regimen consists of AVE9633 IV infusion on Day 1 of a 3 weeks cycle. To date dose levels of 15 (N=3), 30 (N=5), 50 (N=4), 75 (N=4), 105 (N=2), 200 (N=3) and 260 (N=1) mg/m2 have been investigated. Hypersensitivity reactions during perfusion were noted, requiring prophylaxis with steroids. No other AVE9633- attributable extramedullary Grade 3 AE has been observed to date. Free DM4, measured by LC/MS/MS was detectable from the 75 mg/m2 dose level; its Cmax (at the end of infusion) increased from 10 ng/mL at the 75 mg/m2 dose level to 70 ng/mL at 200 mg/m2. Neither AVE9633-associated myelosuppression nor responses have been noted. Using Flow Cytometry Assay on peripheral blasts and monocytes, total saturation and down regulation of CD33 were observed following administration of doses ≥ 30 mg/m2. AVE9633 exposure (measuring, by ELISA method, all antibodies containing at lease one molecule of DM4) increased proportionally with the administered dose in the dose range 15 to 200 mg/m2. Updated PK results and potential explanations for the lack of efficacy using this treatment schedule will be presented.
The Effect of Rituximab in Combination with MEVP Chemotherapy and rhG-CSF in Old Patients of Refractory and Terminal Non-Hodgkin’s Lymphoma.
Authors: Qian Meihua , Wang Wensong , Lan Jianping , Wang Lianming , Zhan Shumei , Zhao Zhonsen , Xu Wenjian 
Published: October 2019

Abstract
It is very important and difficult to study and research the safety and effective methods of treatment in the Terminal or refractory Non-Hodgkin’s Lymphoma ( NHL)patients, especially old patients. To evaluate the safety and efficacy of Rituximab in Combination with MEVP Chemotherapy and rhG-CSF in old patients of Refractory and Terminal Non-Hodgkin’s Lymphoma, 4 old patients of Refractory and Terminal Non-Hodgkin’s Lymphoma( disease history about 6~8 years),3 females and 1 male, between 66~75 years of age (mean age 70.5 years), 2cases IVB, 1 cases IVA, 1case IIIA., Superficial and deep multiple lymph nodes enlarged obviously in all patients and 3 cases of them showed multiple lymph node enlargement and marked lumps in abdominal cavity, 2 cases of them with hydrothorax and ascites, 2 cases of them with bone marrow infiltration and heart disease, 1 case of them with multiple lumps in liver and another of them with autoimmune hemolytic anemia (AIHA), All patients had been treated with chemotherapy included COP (or COPP), CHOP, BCAOP (or BECAOP), MIV (or ESHAP) etc. and 2 patients of them had been treated by radiotherapy before they were treated by Rituximab Combination with MEVP Chemotherapy and rhG-CSF. In this experiment, they were treated by Rituximab plus MEVP Chemotherapy and rhG-CSF,( Rituximab 375 mg/m2/d1, Mitoxantrone 8~10mg/m2/d3, Wumon 0.1/d3~d5 (or VP16 O.1/d3~d7), Vinorelbinum 20mg/m2/d3, Prednison 60mg/d3~d9), rhG-CSF 100~300/d were injected when the number of WBC was <2.0×109/L after chemotherapy. Each cycle was 28~35days. The result is, the minimum of WBC after chemotherapy were 0.6~1.4×109/L, A patient of NHL with AIHA after a treating cycle achieved complete remission (CR), all of enlarged lymph nodes disappeared and Hemoglobin return to normal. Two of patients obtained part remission (PR), most of enlarged lymph nodes disappeared or reduced obviously and hydrothorax and ascites in patients disappeared. The treatment of one patient was inefficacious.
Our study result indicates: The therapy method of Rituximab in Combination with MEVP Chemotherapy and rhG-CSF for the most of patients of Refractory and Terminal Non-Hodgkin’s Lymphoma is safe and effective, no significant toxicity was found during treatment, this treatment method are suitable for most of patients of refractory and Terminal Non-Hodgkin’s Lymphoma and old patients of refractory and Terminal Non-Hodgkin’s Lymphoma.