bluuebunny/crossref_metadata_2025_split huggingface dataset:
Re-Evaluation of Chronic Graft-Versus-Host Disease Using National Institute of Health Consensus Criteria.
Authors: Kim Dae-Young , Lee Je-Hwan , Lee Jung-Hee , Kim Se Hyung , Lim Sung-nam , Lee Young-Shin , Kang Young-Ah , Kang Sol-Ip , Seol Miee , Ryu Seong-Gil , Lee Kyoo-Hyung 
Published: April 2020

Abstract
Background: The National Institutes of Health (NIH) proposed new consensus criteria for the diagnosis and scoring of chronic graft-versus-host disease (cGVHD). Using the new system, we re-evaluated the patients with cGVHD that was diagnosed by classic criteria.
Methods: Of 618 patients who underwent allogeneic hematopoietic cell transplantation (HCT) from December 1994 to April 2008 at the Asan Medical Center, Seoul, Korea, we retrieved 236 patients who had cGVHD by classic criteria from the AMC BMT Registry. Among 236 patients, 20 patients with liver-only involvement could not be diagnosed as cGVHD by the NIH criteria, thus we reclassified and graded 216 patients according to the NIH criteria. We also evaluated the ability of the NIH criteria to stratify and predict the risk of cGVHD patients, as assessed by GVHD-specific survival (GSS).
Results: Twenty patients (9.3%) were reclassified as acute GVHD by NIH criteria (‘classic acute’ in 7 and ‘persistent, recurrent, or late-onset acute’ in 13) and 196 patients (90.7%) remained as chronic GVHD (‘classic chronic’ [Cl-Ch] in 170 and ‘overlap syndrome’ [Ov-Sy] in 26). Median age of 196 patients with cGVHD by NIH criteria, 119 males and 77 females, was 35.5 years (range, 15 to 57). Acute GVHD preceded cGVHD in 70 patients (35.7%). The probability of GSS at 5 years was 86.2% with 22 cGVHD-related deaths. The GSS was significantly different between two subtypes of cGVHD by NIH criteria: 88.6% for Cl-Ch vs 70.2% for Ov-Sy (p=0.002). NIH global scoring system stratified risk of cGVHD patients better than stage by classic criteria at both onset and peak of cGVHD (Table 1). We evaluated 12 variables at onset of cGVHD to determine their prognostic significance for GSS. Multivariate analysis demonstrated that NIH global score at onset (mild vs moderate, HR 6.1, p=0.027; mild vs. severe, HR 7.0, p=0.015), preceding aGVHD (no vs. yes, HR 6.2, p=0.001), and number of HLA ABDR mismatch (0 vs. 1, HR 2.0, p=0.555; 0 vs 2, HR 200.4, p=0.009) were independent predictors for GSS.
Conclusion: Our results indicate that a new NIH system can provide a proper risk-stratification of patients with cGVHD and global scoring system at onset of cGVHD can predict the prognosis of patients.
Table 1. GVHD-specific survival according to NIH global scoring system or stage by classic criteria
Onset of cGVHD Peak of cGVHD Pt. No. GSS (5-y) P-value Pt. No. GSS (5-y) P-value NIH criteria mild 70 95.2% 0.022 36 100% 0.004 moderate 64 82.8% 42 92.2% severe 62 79.7% 118 79.6 Classic criteria limited 86 90.1% 0.305 45 97.8% 0.039 extensive 110 83.7% 151 83.0%
Silencing of Mir-22 Is Linked to H3K27 Trimethylation and Independent of DNA Methylation in Pre-B Acute Lymphoblastic Leukemia Cells.
Authors: Li Xiaoqing , Liu Jun , Zhou Rui , He Yanli , Huang Shi , Huang Shiang , Chen Xian-Ming 
Published: April 2020

Abstract
The regulation of human microRNA (miRNA) expression is still poorly understood and aberrant epigenetic regulation has recently been implicated in the down-regulation of tumor suppressor miRNAs. In this study, we investigated whether histone modifications would contribute to the dysregulation of miRNAs in lymphoblastic leukemia cells. Using a precursor B-cell acute lymphoblastic leukemia cell line, NALM-6 cells, we demonstrated by miRNA microarray analysis that a specific histone deacetylases inhibitor, trichostatin A (TSA), induced a differential alteration in cellular miRNA expression. A total of 10 miRNAs were down-regulated and 31 up-regulated significantly following TSA treatment. Among TSA-up-regulated miRNAs, miR-22 is an extronic miRNA and resides in the second exon of the non-coding transcript MGC14376. Up-regulation of both miR-22 and MGC14376 was found in NALM-6 cells treated with TSA but not 5-AZA-2’-deoxycytidine, a DNA demethylating agent. Luciferase reporter analysis identified three regions in the promoter of miR-22 and MGC14376 that differentially regulated its transcriptional activation. Although there is a CpG island within the promoter of miR-22 and MGC14376, no obvious methylation was detected at this region in NALM-6 cells. Conversely, H3K27 trimethylation (H3K27triM)-associated histone modification was identified in the first intron of MGC14376 gene and was involved in TSA-induced miR-22 expression. Thus, miR-22 silencing in NALM-6 cells involves H3K27triM-associated histone modification but is independent of DNA methylation, suggesting that methylation-independent H3K27triM histone modification may be an important mechanism for miRNA dysregulation in cancer cells.
Acute Myeloid Leukemia Is Characterized by Wnt Pathway Inhibitor Promoter Methylation.
Authors: Griffiths Elizabeth A. , Hooker Craig M. , McDevitt Michael A. , Karp Judith E. , Herman James G. , Carraway Hetty E. 
Published: April 2020

Abstract
The Wnt pathway contributes to a stem-cell like phenotype in a variety of cancer subtypes. Nuclear localization of non-phosphorylated, active β-catenin is a surrogate marker for Wnt pathway activation and has been associated with adverse outcome in patients with acute myeloid leukemia (AML). Wnt pathway inhibitors including APC, DKK1, DKK3, LKB1/STK11, RASSF1A, RUNX3, SFRP1, SFRP2, SFRP4, SFRP5, SOX17, and WIF1 contain extensive promoter region CpG islands. Wnt pathway inhibitors are silenced by promoter methylation in many malignancies including lung cancer, colon cancer and acute lymphoid leukemias. To determine if methylation of these Wnt pathway inhibitors is present in AML, we evaluated leukemia cell lines (i.e., K562, HNT34, KG1, KG1A, U937, and HL60) for evidence of promoter methylation. Additionally, 188 AML patient (median age 61 years) samples from the Johns Hopkins Hospital leukemia tumor bank were assessed for the presence of Wnt pathway inhibitor methylation. All samples were bisulfite treated and evaluated for promoter methylation using methylation specific PCR. Diagnostic samples from a subgroup of patients with normal cytogenetics (n=73) who received high dose induction therapy were evaluated for potential associations between methylation of individual Wnt pathway inhibitor genes or total number of methylated genes and event free or overall survival.
RESULTS: Extensive promoter methylation of the Wnt pathway inhibitor genes was observed in leukemia cell lines. Of the primary leukemia samples, 85% had at least one methylated gene. Promoter methylation of Wnt inhibitors was common in these samples with the following frequencies: DKK1 (16%;30/188), DKK3 (8%;15/188), RUNX3 (27%;50/188), SFRP1 (34%;63/188), SFRP2 (66%;124/188), SFRP4 (9%;16/188), SFRP5 (54%;102/188), SOX17 (29%;54/188), and WIF1 (32%;61/188). This is among the first comprehensive evaluations of Wnt pathway inhibitor methylation in primary samples from AML patients. The frequency of methylation seen here is comparable to that observed in established tumor suppressor genes in patients with AML, including p15INK4B, SOCS1 and CDH1. In marked contrast with epithelial tumors, methylation of APC (2%;2/108) and RASSF1A (0%;0/188) was rare. LKB1/STK11 methylation was also uncommon (2%;2/108). Previous reports have associated Wnt pathway activation with poor prognosis. In our treated patients with normal cytogenetics, no correlation was observed between methylation of Wnt pathway inhibitors and event free or overall survival. In conclusion, in patients with AML (a) there is a high frequency of Wnt pathway inhibitor methylation; (b) Wnt pathway inhibitor methylation is distinct from that observed in epithelial malignancies; and (c) methylation of Wnt pathway genes does not correlate with adverse clinical outcome.
Pathogenesis of Acute Myeloid Leukemia: In Vitro Studies Using Primary Human Cells.
Authors: Abdul-Nabi Anmaar , Yassin Enas R , Takeda Akiko , Yaseen Nabeel R. 
Published: April 2020

Abstract
Many cases of acute myeloid leukemia (AML) are associated with non-random chromosomal rearrangements and most of these result in fusions involving retinoic acid receptor α (RARα), CBF transcription factors, MLL, and nucleoporins. Here, we report the effects of key members of these four major groups of AML-associated chimeric fusion proteins on differentiation, proliferation, self-renewal and gene expression in primary human CD34+ hematopoietic cells. We expressed the PML-RARα, AML1-ETO, MLL-AF9 and NUP98-HOXA9 fusion genes in human peripheral blood CD34+ cells by retroviral transduction and compared them to cells transduced with empty vector. By colony forming assays, morphological examination and flow cytometric immunophenotyping we found that PML-RARα causes some degree of myeloid differentiation block. AML1-ETO had no obvious effect on differentiation, while MLL-AF9 and NUP98-HOXA9 caused a block in both myeloid and erythroid differentiation. All fusion oncoproteins, except PML-RARα, induced dramatic long-term proliferation in liquid culture and a marked increase in the numbers of primitive long-term culture-initiating cells (LTC-ICs). In order to understand the molecular basis of these effects, gene expression profiling was performed for each fusion gene by oligonucleotide microarray analysis at 3 different time points (6 h, 3 days, and 8 days post transduction). At the 6 h time point both AML1-ETO and PML-RARα caused extensive changes in gene expression with a predominance of downregulated genes. The number of genes dysregulated by AML1-ETO and PML-RARα decreased dramatically at the 3-day time point. These data are consistent with the fact that these two oncoproteins are DNA-binding transcription factors that are known to repress transcription. In contrast, MLL-AF9 and NUP98-HOXA9 altered the expression of fewer genes at the 6 h time point with a preponderance of upregulated genes; at the 3 day time point, the number of genes dysregulated by NUP98-HOXA9 and MLL-AF9 increased. These findings suggest that NUP98-HOXA9 and MLL-AF9 may have delayed effects that are not due to direct transcriptional regulation. Homeobox transcription factors were upregulated by both MLL-AF9 and NUP98-HOXA9, but not by either AML1-ETO or PML-RARα. The results of the biological assays and gene profiling show marked similarities between NUP98-HOXA9 and MLL-AF9, and suggest that they transform cells by similar pathways that are different from those used by AML1-ETO and PML-RARα.
NF-κB Is Active in Mcl-1 Promoter Regulation in Human CLL.
Authors: Hertlein Erin K , West Derek A. , Craig Ruth W , Lucas David M , Byrd John C. 
Published: April 2020

Abstract
The NF-κB family of transcription factors is linked to neoplasia due to its role in increasing cell proliferation as well as inhibiting apoptosis. NF-κB has been reported to be constitutively active in chronic lymphocytic leukemia (CLL), but the mechanism underlying this this activation is not fully understood and the critical target genes involved have not been identified. To further define the mechanism(s) by which NF-κB promotes survival in CLL, we investigated the role of this transcription factor in the regulation of the myeloid cell leukemia 1 (Mcl-1) gene. The promoter region of Mcl-1 contains a putative NF-κB binding site, and increased expression of the Mcl-1 protein has been related to rapid disease progression and resistance to apoptosis in CLL. However, a direct link between NF-κB and Mcl-1 transcriptional regulation has not yet been established in B-cells or human CLL. We demonstrate here that NF-κB binding is a critical event in the transcriptional regulation of Mcl-1, as deletion of the NF-κB binding site in the promoter results in decreased activity of an Mcl-1-luciferase reporter construct. In addition, pharmacological inhibition of NF-κB reduced p65 nuclear localization and binding of NF-κB to the Mcl-1 promoter. Interestingly, when CLL patient samples were exposed to an IKK inhibitor, Bay-11, the extent of Mcl-1 inhibition varied in patients. We therefore examined whether the effect on Mcl-1 correlated with in vitro cell survival, a result that might be expected given that NF-κB expression has been previously reported to correlate with CLL cell survival. We found that CLL patient cells more sensitive to Bay-11-induced apoptosis also showed a larger decrease in Mcl-1 mRNA, suggesting that Mcl-1 message level may be a useful diagnostic to predict patients that will respond to NF-κB targeted therapy. Furthermore, different inducers of the NF-κB signaling pathway (CD40L, immune stimulatory CpG-ODN and TNF-α), promote differential effects on Mcl-1 regulation in CLL patient cells. Although all three treatments increase NF-κB nuclear localization and DNA binding, Mcl-1 RNA and protein increased with CpG and CD40L treatment, but not TNFα. This result provides evidence of stimulus-specific regulation of Mcl-1 by NF-κB. Based on these observations, we hypothesize that there is a direct link between NF-κB DNA binding and transcriptional control of Mcl-1, and that this gene could serve as a pharmacodynamic endpoint to monitor the efficacy of NF-κB inhibitors in CLL cells. Additionally, a direct link of NF-κB activity to Mcl-1 expression and cell survival provide further justification for targeting this transcription factor for treatment in CLL.
Impact of Rituximab on Peripheral Blood Stem Cell Mobilization and Collection Following ACVBP Regimen in Poor Risk Patients with Diffuse Large B-Cell Lymphoma (DLBCL): Results from a Large Cohort of Patients from Two Prospective GELA Trials.
Authors: Lefrere Francois , bastit-Barrau Dominique , Mathieu Suzanne , Bohbot Alain , Bourrin Philippe , Salles Gilles Andre , Tilly Herve , Mounier Nicolas , Fitoussi Olivier , Haioun Corinne 
Published: April 2020

Abstract
&lt;&gt;The ACVBP regimen is commonly used in young poor-risk patients with DLBCL candidates to first line consolidative high-dose therapy followed by autologous stem cells transplantation in GELA trials. The combination with the monoclonal anti-CD20 antibody rituximab (R-ACVBP) is now routinely used, as induction treatment and to mobilize peripheral blood stem cell (PBSC). The aim of the present study was to assess the impact of rituximab on PBSC mobilization and collection in patients with newly diagnosed DLBCL receiving ACVBP chemotherapy. We reviewed the data from two prospective controlled trials. The first, conducted between 1999 and 2003, involved patients presenting with 2 or 3 adverse prognostic factors on the basis of the age-adjusted IPI (aa-IPI), treated by ACVBP (LNH 98B-3) (ASCO2007:8018). In the second trial (LNH 03-3B), conducted between 2004 and 2007, similar patients received the same initial inductive chemotherapy combined to rituximab (375mg/m2 at D1). 137 and 91 patients in the ACVBP and the R-ACVBP groups are here analyzed, respectively. Clinical and biological characteristics at diagnosis of the two groups of patients were similar (aa-IPI 2 and aa-IPI 3: 75% and 25%, respectively). The conditions for G-CSF administration and stem cell collections were identical. PBSC mobilizations were performed following the third or fourth cycle of (R)-ACVBP. The median delay between day 1 of chemotherapy and the first hemapheresis was identical for both groups. First hemapheresis was performed with a median peripheral white blood cell concentration of 16.2 x 109/l and 16.6 x 109/l for ACVBP and R-ACVBP groups, respectively. The median peak number of peripheral blood CD34+ cells observed the same day of first hemapheresis in ACVBP and R-ACVBP group was 69 x 106/l and 63 x 106/l, respectively (p = 0.5). The median number of CD34+ cells collected were 7.1 x 106 and 6.0 x 106 CD34 cells/kg for ACVBP and R-ACVBP groups (p = 0.12) while the median number of hemapheresis to target a minimal number of 3 x 106 CD34 cells/kg was identical, of one, in both groups. Failure of stem cell collection , defined as less than 3 x 106 CD34 cells/kg harvested, was observed in 8% and 4% of the patients who received ACVBP or R-ACVBP , respectively (p = 0.13). We conclude that rituximab combined to ACVBP regimen does not impair PBSC mobilization and collection.
The Antioxidant Inflammation Modulator, CDDO-Me Promotes Myelopoiesis in Mice.
Authors: Welniak Lisbeth A. , Harouna Salif , Prosser David , Ames Erik , Meyer Colin , Murphy William J. 
Published: April 2020

Abstract
CDDO-me (methyl-2-cyano-3,12-dioxooleana-1,9-diene-28-oate)is a novel antioxidant inflammation modulator that has been shown to suppress tumor-induced inflammatory processes through suppression of NF-kB and STAT3 and induce apoptosis and differentiation of human leukemic and solid tumor cells.. It has also been shown to be more potent than the parent compound, CDDO against leukemia cells. We have previously shown that administration of CDDO, when given peri-transplant, can inhibit the development of acute murine GVHD and induces cell death in proliferating but not resting lymphocytes in a mixed lymphocyte culture. We therefore next sought to determine the effect of CDDO-me administration on normal hematopoiesis in naive and myelosuppressed mice. Administration of CDDO-me (120 ug/dose BID for 7 days) to C56BL/6 mice did not significantly affect the number of T and B cells in the spleen. Unexpectedly, CDDO-me administration resulted in a significant expansion of myeloid cells in the bone marrow and spleen, predominately CD11b+Ly6CintGR1hi neutrophils. To determine if CDDO-me acted primarily on terminal stages of differentiation or at early phases on development in hematopoiesis, we performed clonogeneic assays for myeloid (CFU-GM), erythroid (BFU-E) and primitive, high proliferative potential (CFU-HPP) precursors and compared treatment with CDDO-me with rhG-CSF administration, a cytokine known to promote expansion and mobilization of hematopoietic precursors from the bone marrow. Administration of CDDO-me resulted in significant expansion of myeloid and erythroid progenitors in the spleen and blood although not to the same extent as that observed in mice receiving 2.5 ug/dose BID rhG-CSF. Unlike the mobilization seen with rhG-CSF where hematopoietic precursors were decreased in the bone marrow, CDDO-me significantly increased the number of myeloid, erythroid and CFU-HPPs in the marrow. Administration of CDDO-me prior to myelosuppression with sublethal total body irradiation also resulted in significant increase of spleen cellularity after 8 days recovery due to significant expansion of myeloid cells as well as increases in B cells. However, CDDO-me did not significantly impact hematopoietic recovery compared to vehicle control treated animals when CDDO-me was given to mice following sublethal total body irradiation. These results suggest that CDDO-me can expand early hematopoietic precursors in mice, resulting in radioprotection. Stimulation of myelopoiesis in vivo by CDDO-me can be accounted for, in part, by a significant increase in circulating G-CSF levels in treated animals. However, the retention of hematopoietic precursors in the bone marrow of mice that received CDDO-me but not rhG-CSF suggest additional mechanisms are involved. These results demonstrate that CDDO-me significantly affects normal hematopoietic cells including precursor populations in vivo and can promote granulopoiesis predominately in non-irradiated mice.
UTY-Coded Minor Histocompatibility Antigens as Targets for a Graft- Versus-Host Reaction against Hematopoietic Cells in the Dog.
Authors: Bund Dagmar , Buhmann Raymund , Zorn Julia , Schmetzer Helga , Kolb Hans- Jochem 
Published: April 2020

Abstract
Female donors produce a strong graft-versus-leukemia (GvL) effect in male patients. Presumably Y-encoded minor histocompatibility antigens (mHA) are recognized by the alloimmune effector lymphocytes. UTY (ubiquitously transcribed tetratricopeptide repeat, Y-linked) is a potential candidate gene for mHA in dogs and men. In the canine model we could show that the minor histocompatibility antigen UTY might be a potential target to further improve the adoptive immunotherapeutic approach. Five HLA-A2 restricted peptides were used for pulsing canine dendritic cells and stimulating canine T cells. T-cells were separated from the blood of female donors using the MACS technique and effector T-cells were generated by incubation of 2x10*6 CD3+ T-cells/ml with autologous mature, peptide-pulsed dendritic cells (DCs) in X-Vivo 15 medium (+ 160 U/ml human IL-2 and 5 ng/ml human IL-7; ratio T:DCs=10:1). T cells were restimulated weekly with peptide-pulsed DCs. After 3 or 4 weeks in culture T-cells were examined for the CD8/CD4 ratio, cytotoxicity assays were assessed after 3–4 re-stimulations in [51Cr]-release assays. The number of peptide-specific T cells was assessed in IFN-g-ELISPOT-assays. UTY-mRNA expression was determined by RT-PCR in several female and male cell types (BM, PBMCs, monocytes, DCs, B-cells). We were able to expand T-cells from 6 female dogs using peptide-pulsed autologous DCs; 3 out of 5 UTY-derived peptides were able to stimulate a specific autologous T-cell response. The number of CD8+ T-cells could be expanded during four weeks of in vitro culture in 3 dogs. In IFN-g-ELISPOT-assays we could demonstrate that the expanded T-cells secrete IFN-g upon recognition of DCs from DLA-identical male littermates. In addition, these T-cells specifically lysed DLA-identical male BM (25-100%), but not autologous or DLA-identical female BM (0–15%) in [51Cr]-release assays (E:T=80:1). Cell types other than BM and DCs (DLA-identical male and female monocytes, PBMCs, B-cells) were not lysed (0–18%). In summary, we could demonstrate that UTY is a naturally processed and presented antigen in the dog. Our data show that HLA-A2 restricted UTY-peptides immunize specific CTLs from female dogs that efficiently recognize DLA-identical male bone marrow and DCs. These findings suggest that UTY may be employed for studies of immunotherapy in the canine model.