bluuebunny/crossref_metadata_2025_split huggingface dataset:
Epigenetic Regulation of the Dendritic Cell Marker Gene MADDAM /ADAM19.
Authors: Kreutz Marina , Ehrnsperger Achim , Rehli Michael , Andreesen Reinhard 
Published: April 2020

Abstract
MADDAM (Metalloprotease And Disintegrin Dendritic Cell Antigen Marker, ADAM19), a human metalloprotease belonging to the ADAM-family, is strongly expressed during in vitro differentiation of monocytes into dendritic cells (DC), whereas differentiation of monocytes into macrophages (MAC) is associated with a loss of MADDAM transcription. To investigate the mechanisms underlying this cell-type specific expression pattern we defined the transcriptional start site and the proximal promoter of the MADDAM gene. Gene bank analysis of the CpG island promoter and first intron revealed putative binding sites for several transcription factors, including VDR, NF-kB and Sp1-family factors. EMSA demonstrated binding of Sp1, Sp3, NF-kB and VDR to their putative binding sites in the proximal promoter region and mutation of these elements led to a decreased reporter activity of the proximal promoter in luciferase assays. A minimal promoter construct of 150-bp showed weak reporter activity in primary monocyte-derived MAC and a threefold higher activity in monocyte-derived DC, indicating that differential binding of transcription factors contributes to the cell-type specific regulation of MADDAM. Transfection of monocytic THP-1 cells with the 150-bp fragment also resulted in significant reporter activity, despite the lack of endogenous MADDAM expression. Interestingly, Trichostatin A (TSA), a known inhibitor of histone deacetylation, lead to a dose dependent induction of MADDAM mRNA in THP-1 cells. Chromatin immunoprecipitation (ChIP) assays demonstrate increased levels of acetylated histones H3 and H4 in DC as compared to MAC, indicating an important role of histone acetylation in the cell-type specific regulation of the MADDAM gene.
Significant Down-Regulation of BAALC during Lineage Specific Hematopoietic Differentiation.
Authors: Baldus Claudia D. , Komor Martina , Hoelzer Dieter , de la Chapelle Albert , Thiel Eckhard , Hofmann Wolf-Karsten 
Published: April 2020

Abstract
The human gene BAALC (Brain And Acute Leukemia, Cytoplasmic) implicated in normal hematopoiesis and leukemia is a marker of immature hematopoietic progenitor cells (PNAS2001;98:13901). To explore its role in hematopoiesis we investigated the regulation of BAALC during lineage specific differentiation. Semi-quantitative RT-PCR was performed on subsets of in vitro differentiated bone marrow CD34+ cells from healthy individuals using the cytokines G-CSF, M-CSF or EPO. Cultures were harvested on day 4, 8, 12, and 16. In addition, oligonucleotide microarray analyses (HG-U133A, Affymetrix) of in vitro stimulated CD34+ cells were independently carried out using EPO, TPO, G/GM-CSF to induce lineage-specific differentiation. For each of the lineages, cells were harvested at days 4, 7 and 11 for expression profiling. All experiments were done in triplicates for each time point and condition. RT-PCR revealed down-regulation of BAALC and CD34 as early as day 4 in cultures with G-CSF, M-CSF or EPO. In concordance, gene expression profiling showed significant down-regulation of BAALC and CD34 on day 4 of culture (BAALC: EPO p=0.04, G/GM-CSF p=0.17, TPO p=0.03; CD34: EPO p=0.01, G/GM-CSF p=0.003, TPO p=0.01). 275 genes showing a significant correlation (correlation coefficient r&gt;0.95) to BAALC expression levels (at the 4 time points and using 3 different cytokines) were identified. Of these CD34 ranked second (r=0.99), and genes related to leukemia and neuronal cells included: Hepatic leukemia factor (HLF; r=0.99) and Paired box gene 5 (PAX5; r=0.99). It has been speculated that BAALC might be involved in cell movement or cell-cell interaction; we therefore investigated the association to genes implicated in cytoskeletal function. Expression of Myosin 5C (MYO5C; r=0.99) and Synaptic nulcei expressed gene 2 (SYNE2; r=0.95) were highly correlated with BAALC expression across all conditions (EPO, TPO, G/GM-CSF). In conclusion, using two independent approaches we show the significant down-regulation of BAALC during hematopoietic differentiation. This underscores the potential role of BAALC in early hematopoiesis and the possible contribution of aberrant BAALC expression in leukemogenesis. We postulate that similar to CD34, PAX, and HLF implicated in hematopoiesis and leukemia, down-regulation of BAALC and structural genes including Myosin 5C are critical steps during hematopoietic differentiation.
New Role of the Regulatory Gene SOX2 in Hematopoiesis.
Authors: Levantini Elena , Bertolotti Francesca , Cerisoli Francesco , Ferri Anna L. , Brescia Elisa , Galvagno Daniele , Tenen Daniel G. , Ottolenghi Sergio , Nicolis Silvia K. , Magli Maria C. 
Published: April 2020

Abstract
Several genes encoding transcription factors of different families have been implicated in the development and differentiation of multiple cell systems. The Sry-type high-mobility-group box 2 gene (Sox2) encodes a transcription factor that is expressed in very early cells such as embryonic stem cells and neural stem cells, where it plays important functional roles (Genes and Dev.17:126, 2003; Development131:3805, 2004). To investigate whether Sox2 plays a role also in blood cell production, we first analyzed its expression in murine hematopoietic cells. Results indicate that the gene is transcriptionally active at low levels in primitive progenitors. Furthermore, in order to address the functional implication of Sox2 in hematopoiesis we analyzed mature and precursor cells in mutant mice compound heterozygotes for a null Sox2 allele and for the deletion of a Sox2 5′ enhancer, as the complete inactivation of the gene in homozygosis is embryonic lethal. At the peripheral blood level we did not detect significant variations in the mutants. However analysis of bone marrow precursors in clonogenic assays showed that Sox2 knock-down mice exhibited a significant increase in the number of multipotent precursors, as compared to wild type animals. Moreover, bone marrow cells of wild type and mutant mice were analyzed for the expression of a panel of regulatory genes involved in the control of different somatic stem cells. Preliminary evidence suggests that some of these genes are modulated in the mutant cells. These observations support the view that Sox2 plays a role at early stages of blood cell production, providing further evidence that common molecular mechanisms may be involved in the regulation of several different types of multipotent cells.
Association of IL-10 and IL-10 Receptor Gene Polymorphisms and Graft-Versus-Host Disease Following Hematopoietic Cell Transplantation.
Authors: Tseng Li-Hui , Lin Ming-Tseh , Storer Barry , Martin Paul J. , Grogan Bryan , Chen Pei-Jer , Zhao Lue Ping , Hansen John A. 
Published: April 2020

Abstract
Polymorphisms in cytokine genes can influence immune responses and may affect the outcome of hematopoietic cell transplantation (HCT). We have shown that the IL-10/-592*A allele of the recipient is a marker for less severe acute graft-versus-host disease (GVHD) and a lower risk of non-relapse mortality (NRM) after HCT from an HLA-identical sibling (N Engl J Med, 2003). To further test the hypothesis that IL-10 pathway is important in the intensity of acute GVHD, we undertook a study of variation in the IL-10 receptor β gene. A single nucleotide polymorphism (A/G) at cDNA position 238 of the IL-10 receptor β gene (IL-10RB/1304) was genotyped in 953 HCT recipients and their HLA-identical sibling donors. IL-10/-592 and IL-10RB alleles and genotypes were tested for association with GVHD by multivariable analysis. The IL-10/-592*A allele of the recipient and IL-10RB/238*G allele of the donor were significantly associated with a lower risk of acute grades III-IV GVHD (trend p value 0.0008 and 0.02, respectively). None of the 16 cases with a patient IL-10 A/A genotype and donor IL-10RB G/G genotype developed grades III-IV acute GVHD (HR = 0.0 and p value = 0.007), compared to pairs with a patient IL-10 C/C genotype and donor IL-10RB A/A genotype. The hazard ratios were 0.4–0.6 among pairs with a patient IL-10 A/A genotype and donor IL-10RB A/G or A/A genotype and among pairs with a patient IL-10 A/C genotype and donor IL-10RB G/G or A/G genotype. The effect of donor IL-10RB genotype on GVHD was observed only among pairs with a patient IL-10 A/C or A/A genotype (trend p value = 0.005 and 0.06 respectively), but not among pairs with a patient IL-10 C/C genotype (trend p value = 0.82). These data suggest an interaction in the effect of the patient IL-10/-592 and donor IL-10RB/1304 genotypes on GVHD, further supporting the hypothesis that the IL-10 pathway plays an important role in controlling the severity of acute GVHD.
Pegfilgrastim after High-Dose Chemotherapy and Autologous Peripheral Blood Stem Cells Transplantation. A Single-Centre Experience in 15 Patients.
Authors: Soussain Carole , Marec Laure , Abarah Wajed , Allard Christian , Mallek Hassina , Itani Corine 
Published: April 2020

Abstract
The efficacy of Pegfilgrastim (PF) in decreasing the duration of neutropenia has been proved after standard dose of chemotherapy. Results of PF after high dose chemotherapy (HDC) and autologous peripheral blood stem cells transplantation ASCT) are lacking. We studied the efficacy of PG in patients receiving HDC and ASCT for lymphoproliferative disease. Fifteen consecutive patients (8 males; 7 females) were onrolled in the study from September 2003 through March 2004. Median age of the patients was 56 years. Diseases were multiple myeloma in 5 patients, diffuse large cell non-Hodgkin’s lymphoma (NHL) in 3 patients, follicular lymphoma in 4 patients, mantle cell lymphoma in one patient, and primary central nervous system lymphoma PCNSL) in 2 patients. All patients were eligible for HDC and ASCT per institutional criteria. Stem cells were collected with peripheral blood pheresis after high dose cyclophosphamide (7 cases); high dose Ara-c (5 cases); ifosfamide (1 case); CHOP-like chemotherapy (1 case); or in steady state (1 case). All the patients received daily G-CSF (5 to 10 mcg/kg). Three different conditioning regimens were used. Patients with multiple myeloma received high dose melphalan (200 mg/m²), patients with PCNSL received a combination of high dose Thiotepa (750 mg/m²), Busulfan (12 mg/kg), and Cyclophosphamide (120 mg/kg), and patients with NHL received a BEAM chemotherapy. PF was administered as a single subcutaneous injection of 6 mg at day +3 after stem cell infusion, except for one patient whose injection was done on day 4. No adverse event attributable to PF was observed. There were no toxic death on study. All patients engrafted neutrophils and platelets. The median time to neutrophils engraftment (&gt; 500/mm3) was 7 days (range, 4–12). Febrile neutropenia was almost constant (14/15) but never exceed OMS grade 2. Median number of days with IV antibiotics was 7 days (range 5–22). These preliminary data show that a fixed dose of 6 mg of PF given subcutaneously at day +3 after HDC and ASCT is safe and effective. A cost efficacy study is warranted to compare PF and daily dose of standard G-CSF after ASCT.
IL-10 Promoter Gene Polymorphism Associated with the Occurrence of Chronic GVHD and Its Clinical Course during Systemic Immunosuppressive Treatment for Chronic GVHD after Allogeneic Peripheral Blood Stem Cell Transplantation.
Authors: Kim Dong Hwan , Lee Nan Young , Sohn Sang Kyun , Baek Jin Ho , Kim Jong Gwang , Lee Jung Lim , Suh Jang Soo , Lee Kun Soo , Lee Kyu Bo 
Published: April 2020

Abstract
Introduction: The current study attempted to evaluate the association between the IL-10 promoter gene polymorphism and transplant outcomes, including the occurrence of chronic GVHD and its clinical course during systemic immunosuppressive treatment (IST) after allogeneic peripheral blood stem cell transplantation (PBSCT).
Methods: Three single-nucleotide polymorphisms in the proximal region of the IL-10 promoter gene (−1082/−819/−592) were analyzed in 60 recipients of cytokine-mobilized PBSCs from HLA-matched sibling donors, and the transplant outcomes compared in terms of the occurrence of chronic GVHD and its clinical course.
Results: The current study only found two haplotypes (1082*A/819*T/592*A [ATA] and 1082*A/819*C/592*C [ACC]). An increased occurrence of chronic GVHD was noted dependent on the IL-10 haplotype (43% vs 68% vs 96% in ACC/ACC vs ATA/ACC vs ATA/ATA haplotype, p=0.003). In a logistic regression based on a multinomial model, the ATA/ATA homozygote presented a 7-fold increased risk of the development of chronic GVHD compared with the ACC/ACC homozygote. The cumulative incidence of chronic GVHD at 1 year post-transplant was 46±20%, 64±10%, and 82±5% in the ACC/ACC, ATA/ACC, and ATA/ATA groups, respectively (p=0.0266). The recipients without the ACC haplotype had a more frequent history of acute exacerbation of chronic GVHD (p=0.031, odds ratio 6.750), while 72% of the recipients with the ACC haloptype did not experience any history of acute exacerbation after the initiation of systemic IST for chronic GVHD compared to 38% of those without the ACC haplotype (p=0.014, odds ratio 4.375). Plus, the duration of systemic IST was significantly shorter in the recipients without the ATA-haplotype compared to those with the ATA haplotype (339 days versus 1,146 days, p=0.0091). In terms of GVHD-specific survival, the recipients without the ATA haplotype showed a 100% survival compared to 70.5% for the ATA haplotype group (p=0.2825).
Conclusion: The IL-10 promoter gene polymorphism was found to be apparently associated with the occurrence of chronic GVHD and its clinical course during systemic IST for chronic GVHD in an allogeneic PBSCT setting.
Figure Figure Figure Figure
Erythropoietin Signaling in Tumor Cells: Differential Activation of Mitogen Activated Protein Kineses and Their Effect upon Cell Survival.
Authors: Udupa Kodetthoor B. , Bose Chhanda 
Published: April 2020

Abstract
Erythropoietin (EPO) regulates the proliferation and differentiation of erythroid progenitors via its receptor, EPO-R and through various Mitogen activated protein kinase (MAPK) pathways. The role of EPO on other cell types is still unkown. In the present study we have elected to examine this aspect in a transformed pancreatic cell line, AR42J cells. We investigated the activation of two MAPKs, namely extracellular regulated kinase (ERK) and c-jun NH2-terminal kinase (JNK) after exposure of AR42J cells to 1 U/ml EPO using the Western blot analysis. We also examined cell viability during EPO exposure by high sensitivity fluoremetric method using CCK8. We found a rapid activation of ERK-1/2 in AR42J cells reaching the maximum of 3.3 fold in 5 min, while it took 30 min for JNK-1/2 to reach the maximum. In the absence of EPO, addition of specific JNK inhibitor, SP600125, reduced cell viability to 50% at a dose of 20 μM while with ERK inhibitor, UO126, cell viability was not reduced even up to 60 μM of the drug. To examine the effect of induction of MAPK by EPO on AR42J cell survival, cells were treated with inhibitors to ERK or JNK 1h prior to EPO addition and the cumulative cell survival were calculated from day 1 through 4. EPO addition to AR42J cells resulted in significantly higher cumulative cell survival of 1.00 ± 0.04 unit absorption compared to the value of 0.35 ± 0.02 unit absorption seen in controls without EPO. When cells were treated with EPO and ERK inhibitor a significantly higher cumulative cell survival of 1.50 ± 0.04 unit absorption was observed (p &lt; 0.01) indicating ERK to be less effective in their survival. On the other hand, samples treated with EPO and JNK inhibitor had significantly lower cell survival (0.55 ± 0.06 unit absorption, p &lt; 0.01) than the EPO control indicating an essential role of JNK in their survival. These results indicate that in EPO mediated survival of AR42J cells, activation of JNK appears to be more important than ERK activation thus indicating a key role of JNK in cell viability. It is very important to understand the role of EPO signaling in the viability of transformed cells since EPO is often supplemented after bone marrow transplantation to enhance the recovery after chemotherapy.
Proliferation Versus Differentiation: The Effect of Growth Factor Stimulation Detected by HALO.
Authors: Rich Ivan N. , Hall Karren , Bartelmez Stephen H. 
Published: April 2020

Abstract
In order for stem and progenitor cells to initiate the proliferation process to grow in vitro and form colonies of differentiated cells, growth factors are required. Some factors are specific for proliferation, some for differentiation. Others exhibit both a proliferative and differentiation function. To date, the classical colony-forming assay (CFA) has been used to ascertain the effectiveness of growth factors either individually or in combination. However, the CFA cannot differentiate between a proliferation or differentiation factor because the assay itself is a differentiation assay. We have now developed a unique in vitro system which measures the proliferative capacity of cells to respond to different growth factors. Based on the colony-forming assay, HALO (Hematopoietic Assays via Luminescence Output) measures cell proliferation as a function of the concentration of intracellular ATP which drives a luciferin/luciferase reaction. The end-point of the reaction is bioluminescence detected in a plate reader luminometer. Since the assay does not require cells to differentiate into colonies, no manual enumeration is needed. Furthermore, HALO is a rapid, non-subjective and standardized proliferation assay performed in 96-well plates with high-throughput capability. At least 14 different cell populations with differing proliferative potential from 5 different species can be detected simultaneously. For the present study, both mouse and human bone marrow target cells were used. Eight replicate microcultures of 100ul each were performed to investigate the dose-dependent effect of single growth factors on cell proliferation. The results are summarized as follows. Proliferation of mouse and human hematopoietic cells can be detected between 1 and 2 days and is optimal between 4 and 7 days respectively. The growth factors, SCF, IL-3, IL-6, Flt3-ligand, EPO, TPO and GM-CSF alone, all induce proliferation at less than 1ng/ml. At growth factor concentrations typically used in the colony-forming differentiation assay, many are inhibitory. The EPO dose response for murine CFU-E proliferation measured at 21h produced a flat response and inhibition at doses above 10mU/ml. The mirror image profile was obtained for CFU-E differentiation at 48 h. These results demonstrate that at the CFU-E level, EPO is a differentiation factor and not a proliferation factor. However, EPO alone does stimulate proliferation at the BFU-E stage of development. In conclusion, it is now possible to distinguish between a proliferation and differentiation factor at different hierarchical levels in the hematopoietic system.
Myeloid Progenitor Cells from Gadd45a and Gadd45b-Deficient Mice Are Sensitized to Genotoxic-Stress Induced Apoptosis.
Authors: Gupta Mamta , Gupta Shiv K. , Balliet Arthur G. , Hoffman Barbara , Lieberman Dan A. 
Published: April 2020

Abstract
GADD45 (Growth arrest and DNA damge) regulates cell growth following exposure to diverse stimuli. It has been shown that, mice lacking the gadd45a gene exhibit genomic instability and increased carcinogenesis, but the exact role of the gadd45 family genes still remains unclear. In this study we have aimed at determining the effect of gadd45a or gadd45b deficiency on the response of bone marrow derived myeloid cells to genotoxic stress agents by using gadd45a or gadd45b null mice. We have found that myeloid progenitor cells from gadd45a or gadd45b-null mice are more sensitive to ultraviolet-radiation (UV), VP-16 or daunorubicin induced apoptosis. Introduction of wild-type gadd45 into gadd45-deficient bone marrow cells restored the wild-type apoptotic phenotype. In-vitro colony formation following stress responses has shown that bone marrow cells from gadd45a or gadd45b-deficient mice have a decreased ability to form haematopoetic colonies. Gadd45a or gadd45b-deficient bone marrow cells also displayed defective G2/M cell cycle checkpoint following exposure to either UV and V-16 but were still able to undergo G2/M arrest following exposure to daunorubicin, indicating the existence of different G2/M checkpoints in response to these anticancer agents. Taken together these findings identify gadd45a or gadd45b as anti-apoptotic gene(s), and suggests that the absence of gadd45a or gadd45b results in higher susceptibility of haematopoetic cells to UV radiation and certain anticancer drugs.
Tumor-Derived Lactic Acid Modulates Dendritic Cell Activation and Differentiation.
Authors: Kreutz Marina , Gottfried Eva , Kunz-Schughart Leoni , Hoves Sabine , Andreesen Reinhard , Muller-Klieser Wolfgang , Mackensen Andreas 
Published: April 2020

Abstract
The tumor milieu can influence DC differentiation in vivo and in vitro. We analyzed DC differentiation in a 3-dimensional tumor model and propose a new mechanism of DC modulation by the tumor environment. Monocytes were cultured in the presence of IL-4 and GM-CSF within multicellular tumor spheroids (MCTS) generated from urothelial carcinoma (J82, UMUC3) and melanoma cell lines (MelIm, Mel108). Monocytes invaded the tumor spheroids and differentiated into tumor-associated dendritic cells (TADC) that displayed an altered phenotype compared to DC generated without tumor contact. The expression of CD1a was reduced on TADC whereas CD80, CD86 and CD16 were upregulated. In addition, TADC generated within J82-MCS showed a reduced expression of HLA-DR and accordingly the antigen presentation in a mixed lympocyte reaction was decreased. We then analyzed the underlying mechanism. Supernatants of MCTS could partially transfer the suppressive effect. Conditioned media from urothelial carcinoma cell lines contained high levels of M-CSF and IL-6, both cytokines known to modulate DC differentiation. In contrast, melanoma MCTS cocultures produced little M-CSF and IL-6, but further analysis revealed that melanoma cells secreted high levels of lactic acid. Indeed, addition of lactic acid during DC differentiation in vitro induced a phenotype comparable to TADC generated within melanoma MCTS. Inhibition of melanoma cell lactic acid production by the addition of oxamic acid, an inhibitor of lactate dehydrogenase, reverted the TADC phenotype back to normal. We therefore conclude that tumor-derived lactic acid is an important factor modulating the DC phenotype in the tumor environment which may critically contribute to tumor escape mechanisms.