bluuebunny/crossref_metadata_2025_split huggingface dataset:
Total Therapy 2 (TT2) for Multiple Myeloma (MM): Thalidomide (T) Effects Superior Complete Response (CR) and Event-Free Survival (EFS); Similar Overall Survival (OS) Linked to Shorter Post-Relapse Survival.
Authors: Tricot Guido , Rasmussen Erik , Anaissie Elias , van Rhee Frits , Zangari Maurizio , Hollmig Klaus , Jackson Lisa , Epstein Joshua , Shaughnessy John , Crowley John , Barlogie Bart 
Published: April 2020

Abstract
Background: Autotransplants have become the standard of care for patients with MM. TT2, a prospective randomized phase III trial, evaluated whether adding T, effective in end-stage MM, could improve the outcome of patients receiving melphalan (MEL200)-based tandem autotransplants.
Methods: Between October 1998 and February 2004, 668 patients were randomized to receive T (n=323) or no T (n=345), with the primary objective to increase the 5-yr EFS rate from 40% to 50%. TT2 consisted of 4 phases: induction with VAD-DCEP-CAD with PBSC collection, DCEP; tandem autotransplants with MEL 200; D-PACE consolidation; interferon maintenance with added DEX pulsing during the 1st year. The median follow-up is 3 years.
Results: Patient baseline characteristics in the 2 arms were similar. T increased CR from 41% to 59% (p&lt;0.001) (Figure 1a); 5-yr EFS was superior for the T arm (54% vs. 42%, p=0.017), whereas 5-yr OS was not different at 67% and 62% (p=0.9), due to shorter post-relapse survival of patients initially randomized to T (13 vs 32mos, p&lt;0.001) (Figure 1b). Of 437 patients with interphase FISH data, amp1q21 and chromosome 13q14 deletion (del13q14) both conferred short OS (p&lt;0.001, p=0.003) and EFS (p&lt;0.001, p&lt;0.001), independent of metaphase cytogenetic abnormalities (CA) (p&lt;0.001); these 3 genetic features alone accounted for 35% of OS and 28% of EFS variability. T improved EFS both in patients without and with one or more of these 3 genetic abnormalities. High amp1q21 indices accounted for shorter post-relapse survival (p&lt;.001), independent of initial T randomization (p=.5). The post-relapse therapies were similar among the 2 arms and included T in all previously not on T and most previously on T, often together with bortezomib = DEX (VTD regimen; Barlogie, Blood, 2004).
Conclusions: Intensive chemotherapy before and following tandem autotransplants in TT2 effected unprecedented high 5-yr EFS (48%) and OS rates (65%). T significantly increased CR frequency and extended EFS but not OS. A shorter post-relapse survival could be traced to high amp1q21 index at relapse (more often present on T arm).
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Bacterial Endotoxin LPS but Not LTA Can Enhance Osteogenic Differentiation of Human Mesenchymal Stem Cells.
Authors: Chan Godfrey ChiFung , Mo F.Y. , Yip K.H.K. , Li J. , Law H. , Lau Y.L. 
Published: April 2020

Abstract
Background & Objective: Dental implant requires osseointegration for anchoring and human’s oral cavity has plenty of bacterial oral flora. Whether these bacteria have any effects on the human mesenchymal Stem Cells (MSCs) that can differentiate into osteoblasts remains unknown. We therefore investigated the effect of bacterial endotoxins commonly found in the oral cavity and gastrointestinal tract, namely lipopolysaccharides (LPS, Escherichia coli) and lipoteichoic acid (LTA, Streptococcus pyogenes), on the proliferation and osteogenic differentiation of MSCs.
Methods: Human MSCs are derived from bone marrow (BM) of normal healthy donors. The culture condition, immunophenotyping determination and tests of differentiating functions of the human MSCs were similar to what we reported previously (Li J, Br J Haematol 2004). The proliferation of MSCs under either a 3-day or a prolonged 7-day endotoxins challenge was evaluated by XTT assay. The extent of osteogenic differentiation was examined under microscopy and measured by the increase in alkaline phosphatase (ALP) activity at day 10 and the calcium mineralization/deposition per unit volume of protein at day 14.
Results: There was no significant effect of LPS and LTA on the growth and proliferation of MSCs, even under a relatively high dose. However, continued LPS challenge on MSCs under osteogenic culture condition was shown to increase the ALP activity and calcium deposition in a dose-dependent manner (100ng/ml, 1 ug/ml, 10ug/ml). No such phenomenon can be identified when LTA challenge was used.
Conclusions: LPS and LTA did not show any significant effect on the proliferation and growth of human MSCs. However, LPS enhanced the osteogenic differentiation of MSCs in a dose-dependent manner. Our finding suggests that the endotoxin from bacteria commonly found in the oral cavity and gut does not have any negative impact on MSCs induced osteogenesis.
Mobilization of Stem Cells from the Bone Marrow Is Required for Neovascularization of Ischemic Limbs in Mice.
Authors: Kim Jeong-A , Lee Chang -Hoon , Yoon Jin-A. , Min Woo-Sung , Kim Chun-Choo 
Published: April 2020

Abstract
We examined whether the injection of bone marrow mononuclear cells (BM-MNCs) or mesenchymal stem cells (MSCs) might augment angiogenesis and collateral vessel formation in a mouse model of hind limb ischemia. C57BL/6 BM-MNCs were isolated by centrifugation through a Histopaque density gradient and MSCs were obtained from C57BL/6 bone marrow and cultured in low-glucose DMEM media. Unilateral hind limb ischemia was surgically induced in C57BL/6 mice (control; n=4), and autologous BM-MNCs (Group 1; n=4, 1.8±0.2 x107/animal) or MSCs (Group 2; n=4, 1.0±0.14 x106/animal) or BM-MNCs and MSCs (Group 3; n=4, 2.3±0.1 x107 and 1.1±0.21 x106/animal) were transplanted into the ischemic tissue. Six weeks after transplantation, the group 1, group 2 and group 3 had a higher capillary/muscle ratio (0.82±0.12 vs 0.85±0.08 vs 0.97 ±0.03) than control (0.46±0.12, p&lt;0.05) (Fig. 1). This result suggested that direct local transplantation of autologous BM-MNCs or MSCs seems to be a useful strategy for therapeutic neovascularization in ischemic tissues.
Next, we evaluated whether bone marrow derived stem cells were participated in the process of local injected stem cells forming new vessels. In general, mobilizing stem cells from bone marrow to local site, MMP-9 has been known as an important molecule. So we used the MMP-9 deficient KO mice and wild type, 129SvEv mice were used in the experiments. Autologous BM-MNCs and MSCs were transplanted into the ischemic limb in MMP-9 (−/−) (n=4) after unilateral hind limb ischemia was surgically induced and then the same experiments was done in MMP-9 (+/+) mice (n=4). The number of the injected BM-MNCs and MSCs was 2.2±0.05 x107 and 0.87±0.17 x106/animal in MMP-9 (−/−). And the number of the injected BM-MNCs and MSCs was 2.1±0.17 x107 and 0.98±0.09 x106/animal in MMP-9 (+/+). No difference was seen in the BM-MNCs and MSCs were injected or not (0.52±0.07 vs 0.49±0.03,) in MMP-9 (−/−). But, in the case that BM-MNCs and MSCs were injected, the higher capillary/muscle ratio was seen in MMP-9 (+/+) compared to control (0.86 ±0.09 vs 0.49±0.03, P&lt;0.05) (Fig 2). This data indicated that the mobilization of bone marrow derived stem cells would have an important role in the neovasculrization although the stem cells were injected directly into the muscle of ischemic limb.
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Aberrant DNA Methylation of the C/EBPalpha Promoter Region in Hematopoietic Malignancies.
Authors: Jost Edgar , do Nicole , Herman James G. , Osieka Rainhardt , Galm Oliver 
Published: April 2020

Abstract
The CCAAT/enhancer binding protein alpha (C/EBPalpha) is a transcription factor implicated in myeloid differentiation. Loss of function of C/EBPalpha may impair granulocytic differentiation and thus contribute to leukemogenesis. Inactivation mutations of C/EBPalpha have been reported in hematopoietic malignancies, especially in acute myeloid leukemia (AML). Hypermethylation of CpG islands within gene promoter region is associated with transcriptional inactivation and represents an important mechanism of gene silencing in hematological malignancies. This epigenetic phenomenon acts as an alternative to mutations and deletions to disrupt gene function. In this study, we have analyzed the methylation status of the promoter-associated CpG island of the C/EBPalpha gene in hematopoietic cell lines and primary blood and bone marrow samples from patients with AML, multiple myeloma (MM) and non-Hodgkin’s lymphoma (NHL) by methylation-specific polymerase chain reaction (MSP). C/EBPalpha promoter methylation in L540 cells was associated with transcriptional repression, and treatment with the demethylating 5-aza-2′-deoxycytidine resulted in C/EBPalpha upregulation. MSP analysis of primary patient samples revealed C/EBPalpha hypermethylation in 19.3 % (11/57) of AML, 27.1 % (13/48) of MM and 46.7 % (7/15) of NHL. Aberrant C/EBPalpha methylation was found in AML of all FAB subtypes and throughout all cytogenetic risk groups. In MM, C/EBPalpha hypermethylation occurred in all Salmon and Durie stages as well as in plasma cell leukemia. Our data indicate that hypermethylation of the transcription factor C/EBPalpha is a common finding in various hematopoietic malignancies and may contribute, in addition to genetic aberrations, to the malignant phenotype. Our growing knowledge about epigenetic aberrations not only gives further insight in the pathogenesis of hematopoietic malignancies but also provides a rationale and molecular basis for therapeutic approaches with demethylating agents.
A Single Nucleotide Polymorphism and Its Haplotype of ZNF42 Transcriptional Modulator Gene Predisposes Individuals to High-Risk Acute Myelogenous Leukemia (AML).
Authors: Lee Il-Kwon , Choi Jeong-Hwa , Lee Yu Ra , Kim Yeo-Kyeoung , Kim Hee Nam , Park Kyeong-Soo , Lee Je-Jung , Shin Myung Geun , Choi Chan , Kook Hoon , Hwang Tai Ju , Park Moo Rim , Jo Deog Yeon , Won Jong Ho , Kim Hyeoung-Joon 
Published: April 2020

Abstract
ZNF42 (MZF-1 or Human Zinc finger protein 42) is a hematopoietic transcription factor regulating the differentiation and proliferation of myeloid cells. ZNF42 belongs to the Krupple-class zinc finger protein, which is expressed in myeloid progenitor cells. As a transcriptional regulator it has been found to be pivotal in hematopoietic development, especially in granulopoiesis. ZNF42 is a bi-functional transcriptional regulator, by repressing transcription in non-hematopoietic cells, but trans-activating hematopoiesis-related genes. Despite the important role of ZNF42 in differentiation and proliferation in the myeloid lineage cells, molecular epidemiologic studies of ZNF42 have not been conducted. In this case-control study, we conducted a comprehensive analysis of the genomic region of ZNF42, including SNP discovery by re-sequencing of the promoter and exon-encompassing regions, haplotype construction and construction of linkage disequilibrium (LD) map and comparison LD structure among four different populations data from the International HapMap project. In total, 275 de novo AML patients plus age- and sex-matched controls were recruited and four coding non-synonymous SNPs were genotyped by Pyrosequencing for this study. All genotypes frequencies were in Hardy-Weinberg equilibrium. A non-synonymous SNP (G/A altering amino acid R51H) revealed strong association with increased susceptibility to AML. Relative to individuals with the GG genotype, those with the A allele (AA + AG) had a 4.8-fold (95% CI, 3.3–7.0) (p=4.111e-17) risk of development of AML. Also four common (&gt; 5% frequency, cumulative frequency of over 96%) haplotypes were identified and the frequencies of the common haplotypes predicted were similar between cases and controls. When we studied if there is haplotype-based association to examine the contribution of common genetic variation at the ZNF42 locus to AML risk among Korean, two haplotypes (G-G and A-G from R51H-R130Q) showed statistically significant association.. These results suggest that ZNF42 variants and its haplotypes are significantly associated with increased susceptibility to AML.
Lineage-Specific Expression of Mouse Abcg2 during Hematopoiesis Is Regulated by Alternative Use of Multiple Leader Exons and Promoters.
Authors: Zong Yang , Zhou Sheng , Sorrentino Brian P. 
Published: April 2020

Abstract
ABCG2 encodes a transmembrane transporter associated with multi-drug resistance in various cancer cells. ABCG2 is also highly expressed in hematopoietic stem cells (HSCs) and is down-regulated in most committed progenitors, while expression is sharply up-regulated during erythroid differentiation and significant levels of protein expression are detected on mature red blood cells. The expression of ABCG2 in these hematopoietic cells likely reflects a protective role against both endogenous and exogenous toxins. The mechanisms for regulation of ABCG2 expression in hematopoietic cells are poorly understood. We have recently identified three novel leader exons (termed E1a, E1b and E1c) located in the 5′ untranslated region (5′-UTR) of mouse Abcg2 mRNA by EST database searches and reverse-transcription PCR (RT-PCR). In genomic DNA, the distance of these noncoding exons from exon 2, which contains the translation start codon, is 58.5 kb for E1a, 15.0 kb for E1b and 5.1 kb for E1c. In a mouse erythroid cell line (MEL), RT-PCR analysis showed that the transcript containing E1b exon was the only isoform detected. Furthermore, in primary Ter119+ erythroid cells from mouse bone marrow (BM), real time PCR showed that expression levels of the E1b-containing transcript were at least 100 fold greater than those of the E1a transcript. For comparison, cDNA was isolated from c-Kit+, Sca-1+, Lin- (KSL) BM cells, a population devoid of erythroid cells but highly enriched for repopulating HSCs. Real-time PCR showed that the E1a-containing transcript was the major expressed isoform in KSL cells, while the E1b transcript was present at significantly lower levels. The differential expression pattern of Abcg2 mRNA isoforms in HSCs and erythroid cells indicates that at least 2 different promoters control Abcg2 transcription during hematopoiesis. We also analyzed the human EST database and found 3 different ESTs all containing exon 2, similar to mouse Abcg2 gene. RT-PCR analysis of cDNA derived from human placenta and BM mononuclear cells showed expression of all 3 isoforms, which was confirmed by plasmid cloning and sequencing. These results indicate that the human ABCG2 locus also has three leader exons which are alternatively used and give rise to three isoforms of ABCG2 mRNA that differ in their 5′-UTR. We are currently defining the expression pattern of these isoforms in different human hematopoietic cell subpopulations. In summary, our data show the expression of Abcg2 during hematopoiesis is regulated at the level of transcription by alternative use of multiple noncoding leader exons in lineage-specific manner. Functional analysis of different putative promoter regions is underway and should provide a better understanding of the developmental regulation of Abcg2 expression in hematopoiesis.
A Role for Twist in Thymocyte Development.
Authors: Sugiyama Daisuke , Sugiyama Hiroko , Speck Nancy A. 
Published: April 2020

Abstract
Twist-1 and Twist-2 are basic helix-loop-helix (bHLH)- containing transcription factors important for mesoderm and muscle formation, and for osteoblast differentiation. The Twist proteins were previously shown to interact both physically and genetically with Runx2, a member of the core binding factor family of transcription factors (Bialek et al. Dev. Cell6: 423, 2004). Twist-1 and Twist-2 transiently inhibit Runx2 function during bone formation by inhibiting DNA binding by the Runx2 Runt domain. Since the Runt domains of the core binding factors are highly conserved, this raised the possibility that the Twist proteins may also regulate Runx1 function in hematopoiesis. To address this hypothesis we used Charlie Chaplin (CC) mice, which contain a hypomorphic Twist-1 allele generated by N-ethyl-N-nitrosourea (ENU) mutagenesis (Justice, Nat. Rev. Genet.1: 109, 2000). The CC allele encodes a single amino acid substitution (Ser192Pro) in the Twist box of Twist-1, which normally interacts with the Runt domain. CC/CC mice die from an unknown cause shortly after birth, and have small thymuses. No other hematopoietic organs show significant differences in cellularity. The number of bone marrow hematopoietic progenitors is almost identical among CC/CC, CC/+, and +/+ littermates as assessed by flowcytometry and colony forming assays. The ratios of CD4/CD8 single positive cells, and DN1-4 thymocytes are unperturbed, suggesting that T cell maturation is normal in CC/CC fetuses. However, CC/CC thymuses are reconstituted by wild type bone marrow at a lower efficiency than are wild type thymuses in fetal thymus organ culture assays, suggesting that Twist-1 may have an important non-cell autonomous role in thymocyte development.
Complex Regulation of the Homeobox Transcription Factor TGIF: Differential Expression of Multiple 5′ Isoforms.
Authors: Hamid Rizwan , Patterson Johnequia , Martincic Danko , Brandt Stephen J. 
Published: April 2020

Abstract
TG-interacting factor (TGIF) is a transcriptional repressor belonging to the TALE (three amino acid loop extension) class of homeobox proteins. In addition to its involvement by mutation or deletion in the inherited craniofacial disorder holoprosencephaly, we have shown that expression of TGIF is highly predictive of relapse and survival in acute myelogenous leukemia (AML). To better understand how TGIF expression is regulated, we characterized its genomic structure using expressed sequence tag analysis, reverse transcriptase-coupled PCR, and rapid amplification of cDNA ends. These studies revealed a complex pattern of gene expression, with 15 splice isoforms and 11 alternative 5′ exons spread over 40 kb of DNA, suggesting the presence of multiple promoters. Real-time and semi-quantitative PCR analysis showed these isoforms were differentially expressed in various adult tissues, leukemic cell lines, and AML blasts. Further, isoform C was found to be the major RNA product in hematopoietic cells, contributing significantly to total TGIF expression in leukemic cell lines TF1a, U937, AML-193, KG-1a, Kasumi-1, K562, GDM-1, HL-60 and AML blasts. This analysis suggests that altered splicing and/or expression of specific isoforms could be responsible for the reduced levels of TGIF message observed in AML blasts and cell lines. The unusually complex structure of the TGIF gene may enable its precise regulation during normal hematopoiesis and may be relevant to its reduced expression in myeloid leukemias.
A Novel Switch Culture System for Proliferation and Differentiation of Monocytic Cells: ERK Activation Is Not Sufficient for Macrophage Differentiation by M-CSF.
Authors: Suzu Shinya , Harada Hideki , Kimura Fumihiko , Motoyoshi Kazuo , Okada Seiji 
Published: April 2020

Abstract
M-CSF, a ligand for Fms, is an essential cytokine for proliferation and differentiation of monocytes/macrophages. However, molecular events responsible for M-CSF/Fms-mediated differentiation and their differences to proliferation-inducing signals are poorly understood. To clarify the difference, it is required to establish cell culture systems, in which distinct responses (proliferation versus differentiation) can be assessed independently in same cells. Here, we report the establishment of a novel culture system useful for analyzing the differences. TF-1-fms leukemia cells proliferate in response to M-CSF, but tend to differentiate into macrophages in the presence of a phorbol ester TPA. Interestingly, in the presence of TPA, M-CSF was likely to hardly yield the initiation of proliferation signal but markedly accelerate the TPA-triggered macrophage differentiation process. The combination of both TPA and M-CSF caused more drastic morphologic change than the treatment with TPA alone. The cells cultured with TPA and M-CSF showed increased granularity and higher phagocytic activity than those cultured with TPA alone. However, there was no significant difference in the numbers of viable cells and in the percentage of PI-positive dead cells between the two treatments, suggesting that the differentiation-accelerating effect did not reflect the survival-enhancing activity of M-CSF but the factor actively transmitted differentiation signals to TF-1-fms in the presence of TPA. The differentiation by TPA/M-CSF apparently correlated with elevated phosphorylation of ERK, a possible regulator of differentiation in a number of cell systems. However, and of importance, a specific MEK inhibitor, PD98059, inhibited the ERK phosphorylation but not macrophage differentiation. The result raised the possibility that elevated ERK activation was not necessarily sufficient for macrophage differentiation of TF-1-fms cells mediated by M-CSF. The experiments with TF-1-fms cells expressing HIV-1 Nef protein further supported the hypothesis: TF-1-fms cells over-expressing Nef showed impaired macrophage differentiation response but increased ERK phosphorylation (Suzu S et al., Blood 2005). Therefore, our switch culture system provides a useful tool for identifying molecules regulating the M-CSF/Fms-mediated macrophage differentiation.
Leptin-Induce Proliferation Mediated by JAK/STAT and MAPK Activation in Acute Myelogenous Leukemia Cells.
Authors: Park Hyun Ki , Kim Ju Young , Yoon Jin Sun , Kim Eun Shil , Ahn Kwang Sung , Yoon Sung Soo , Kim Byoung Kook , Lee Young Yiul 
Published: April 2020

Abstract
Leptin, secreted as a product of the ob gene that is mainly produced by adipose tissue, has been involved in the regulation of energy metabolism. Recently, leptin has been suggested to stimulate normal cells as well as various cancer cells including acute myelogenous leukemia (AML). However, the molecular mechanism of leptin as a proliferative effector is not poorly understood in AML. In the present study, we show that leptin-induced cellular proliferation is mediated by JAK/STAT and MAPK activation in AML. Expression of the long form (Ob-Rb) and the short form (Ob-Ra) of leptin receptor (Ob-R) was observed in 10 AML cell lines examined, and up-regulated by treatment of leptin. Leptin induced the proliferation of HEL, which was shown to express the highest Ob-R among AML cell lines, in a dose-dependent manner. Treatment with 100ng/ml of leptin enhanced the expression of Janus kinase 2 (JAK2) phosphorylation, signal transducer and activator of transcription (STAT)-3 phosphorylation and mitogen-activated protein kinases (MAPKs) in HEL. Blocking of STAT3 phosphorylation with a specific inhibitor, AG490 (50 μM), significantly reduced leptin-induced ERK1/2 phosphorylation and cellular proliferation of HEL, whereas blocking of ERK1/2 activation by a specific ERK1/2 kinase inhibitor, PD98059 (25 μM), did not affect the STAT3 phosphorylation and leptin-induced proliferation in HEL. Furthermore, knockdown of Ob-R expression with small interfering RNA (siRNA) reduced leptin-induced proliferation of HEL, and also significantly attenuated leptin-induced STAT3 and ERK1/2 activation. This results provides that leptin promotes AML cell growth by activating JAK/STAT3 and MAPK, although not directly dependent on ERK. Blocking as direct receptor level could be a rational therapeutic strategy of AML.