bluuebunny/crossref_metadata_2025_split huggingface dataset:
GITR Ligand on Leukemic Myeloid Dendritic Cells Suppresses Induction of Leukemia-Associated Antigen-Specific CTLs from Naïve CD8+ T Cells.
Authors: Kondo Yukio , Katagiri Takamasa , Ohata Kinya , Nakao Shinji 
Published: April 2020

Abstract
Glucocorticoid-induced TNFR-related protein (GITR), a member of the TNF receptor superfamily, is expressed at low levels on resting T cells and is up-regulated following activation. Triggering of GITR with its ligand (GITRL) has been described to abrogate the function of CD4+CD25+ regulatory T cells and stimulate effector T cells in mice, but little is known about roles of GITR-GITRL interaction in humans. Recent evidence suggests that the interaction between GITR on NK cell and GITRL, which is constitutively expressed on tumor cells, negatively regulates NK cell-mediated anti-tumor activity in cancer patients. Leukemic myeloid dendritic cells (mDCs) can induce cyclin-dependent kinase (CDK2)- specific cytotoxic T lymphocytes (CTLs) from naïve T cells and CDK2-specific CTLs are detectable in MRD+ patients with leukemia after allo-SCT (Blood. 110 (11): a3228. 2007). The GITR-GITRL interaction may affect this process and a modification of the interaction may improve the efficiency of inducing CDK2-specific CTLs. Therefore, the expression of GITRL on leukemic cells and leukemic mDCs was examined to determine whether an engagement of GITR controls the priming of leukemia-associated-antigen (LAA)-specific CD8+ T cells. When cryopreserved BMMCs obtained at diagnosis from 5 patients with AML were assessed using flow cytometry, GITRL expression was detectable on leukemic cells in 3 patients. Leukemic mDCs were enriched from PBMCs of HLA-A24+ patients with anti-CD1c mAb-conjugated magnetic beads and were assessed for their ability to stimulate HLA-A24+ naïve CD8+ T cells to acquire cytotoxicity specific to CDK2-peptides (CDK2 158–166, CDK2 178–186). A three days culture of immature leukemic mDCs in the presence of TNFα up-regulated the expression of GITRL along with CD83 and CD40 expression. Naïve CD8+ T cells isolated from healthy individuals and cord blood were cultured with the GITRL-expressing leukemic mDCs in the presence or absence of anti- GITR monoclonal antibodies (mAbs) for 14 days and stained for CDK2 158–166/A24, CDK2 178–186/A24 pentamers. Blocking of GITR with the mAbs augmented induction of CDK2 158–166- and CDK2 178–186- specific CD8+ T cells from 0.37% to 1.17% and from 0.45% to 1.64%, respectively (Fig). Anti-GITR mAbs did not enhance induction of CDK2-specific T cells by peptide-pulsed monocyte-derived DCs which do not express GITRL. These data suggest that the expression of GITRL on circulating leukemic mDCs may suppress induction of CTLs specific to LAAs and induce cancer immunoediting in patients with leukemia. Administration of anti-GITR mAb after allo-SCT may enhance graft versus leukemia effect by CDK2-specific CTLs without vaccination of CDK2-peptides.
Fig Blocking of GITR on T cells augments induction of CDK2-specific CTLs Fig. Blocking of GITR on T cells augments induction of CDK2-specific CTLs
Indirect Comparison of the Efficacy of Melphalan-Prednisone-Bortezomib Relative to Melphalan-Prednisone-Thalidomide and Melphalan-Prednisone for the First Line Treatment of Multiple Myeloma
Authors: Yeh Yating , Chambers James , Gaugris Sabine , Jansen Jeroen 
Published: April 2020

Abstract
Melphalan-prednisone (MP) combination has been considered a standard of care for front line treatment of multiple myeloma in patients non eligible for transplant. Melphalan-prednisone-bortezomib (MPV) combination has been approved in the United States in patients non eligible for high-dose chemotherapy (HD-C) and has recently received a positive opinion from the CHMP in Europe. Melphalan-prednisone-thalidomide (MPT) was approved in Europe in patients &gt;65 or not eligible for HD-C. There is no head-to-head trial directly comparing MPV to MPT. The objective of the current study was to compare the efficacy of MPV to MP and MPT as first line treatment of multiple myeloma in patients non eligible for transplant. Six randomized placebo controlled trials investigating the efficacy of MPT (5) and MPV (1) relative to MP were identified with a systematic literature review. The endpoints of interest were overall survival (OS), progression free survival (PFS) and overall and complete response. Relative efficacy estimates of MPT versus MP as obtained from the MPT-MP trials were combined with meta-analysis techniques and simultaneously indirectly compared with the relative efficacy of MPV versus MP from the MPV-MP trial (VISTA). This adjusted indirect comparison was performed with Bayesian fixed and random effects models. As compared to frequentist approach, Bayesian meta-analysis offers a more informative summary of the likely value of efficacy after observing the data and allows for direct probabilistic inferences. Of the three interventions compared, there was an 81% probability that MPV was the most efficacious intervention in terms of overall response and a &gt;99% probability in terms of complete response. With MPV a patient was two times more likely to show a complete response than with MPT (Relative Risk=2.15; 95%Credible Interval (CrI): 0.99–4.45). Both MPV and MPT showed greater OS than MP (HR=0.61; 95%CrI: 0.42–0.88 and HR=0.61; 95%CrI: 0.47–0.78 respectively); the indirect comparison showed similar efficacy in terms of OS between MPV and MPT (MPV vs MPT: Hazard Ratio=1.00; 95%CrI 0.64–1.57). Both MPV and MPT also displayed greater PFS than MP (MPV versus MP: HR=0.61; 95%CrI 0.49–0.76 and MPT versus MP HR=0.51; 95%CrI 0.41–0.63 respectively) and showed similar efficacy (MPV vs MPT: HR=1.19; 95%CrI: 0.87–1.63). In this study, both MPV and MPT are more efficacious than MP in terms of response, PFS and OS. MPV is expected to result in a greater complete and overall response than MPT. No difference in OS or PFS was displayed. Further analyses will need to be undertaken once evidence base data is more mature.
Quality of Life Scores Pre and Post Autologous Stem Cell Transplant: A Comparison of Patients Greater Than 60-Years-Old Vs Younger Than 60-Years-Old
Authors: Dabney Jane , Rybicki Lisa , McLellan Linda , Foster Larry , Visnosky Michele , Curtis Julie , Bolwell Brian 
Published: April 2020

Abstract
Prospective quality of life instruments were administered to 304 patients with non-Hodgkin’s lymphoma (NHL), Hodgkin disease (HD), and Multiple Myeloma (MM) undergoing autologous stem cell transplantation (ASCT) from June 2003 through December 2007. The two groups were fairly balanced with respect to baseline characteristics with the younger group having slightly more HD and the older group having slightly more NHL. Of the 304 total patients, 65% had NHL, 21% MM, and 14% HD; median age was 53; 60% were male; 25% received prior radiation therapy; preparative regimens were busulfan based or melphalan; all patients received peripheral stem cells alone, median inpatient length of stay was 21 days. The Functional Assessment of Cancer Therapy-Bone Marrow Transplant (FACT-BMT) and the Profile of Mood State (POMS) were administered to patients prior to their priming for stem cell collection and again 40–44 days (Day 42) post ASCT. The FACT-BMT measures five components: physical well being (PWB), social well being (SWB), emotional well being (EWB), functional well being (FWB), and additional concerns (AC). The POMS measures depression, vigor, anger, tension, confusion, and fatigue. In looking at the FACT-BMT the 60 + group reported better emotional well-being at baseline than the &lt;60 group (p= 0.008) and on day 42 post transplant the 60+ group reported significantly better EWB from the under 60 group (p&lt;0.001). Both groups report a decrease in FWB from baseline to day 42 (p=0.022) but the 60+ group reports higher FWB than the other group. Results of the POMS indicates that the &lt;60 group reported higher depression and anger at baseline than did the 60+ group (p&lt;0.001). The &lt;60 group also reported greater tension (p=0.002) at baseline. Confusion was rated slightly higher at baseline also by the &lt;60 group (p=0.020). There were no significant differences between the two groups in these areas at day 42. In conclusion, one can speculate that the 60+ group is more likely to have experienced family or personal crisis prior to their diagnosis and may have more established coping mechanisms on which to rely. Additionally, those in the &lt;60 group may tend to have more family and employment considerations that may distract them from being able to focus on their treatment and preparation for transplant. It is possible that the &lt;60 group would benefit from earlier involvement from the social worker to assess their needs and more specialized family oriented education and/or materials.
Granulocyte Colony-Stimulating Factor (G-CSF) Stimulates Sympathetic Nervous System Activity in the Bone Marrow
Authors: Battista Michela , Mendez-Ferrer Simon , Frenette Paul S. 
Published: April 2020

Abstract
Granulocyte colony-stimulating factor (G-CSF) is the agent most commonly used clinically to elicit hematopoietic stem and progenitor cells (HSPCs) mobilization from the bone marrow (BM) to the peripheral blood. Our previous studies indicate the sympathetic nervous system (SNS) controls G-CSF-induced HSPCs egress from the BM and that G-CSF decreases catecholamine levels in bone/BM tissues (Cell2006; 124:407). However the molecular basis for this depletion is unknown. Here we show that G-CSF stimulates selectively the activity of the SNS in bone/BM microenvironment in mice. Norepinephrine (NE), a catecholamine and the principal neurotransmitter of the SNS, is synthesized by the rate-limiting enzyme tyrosine hydroxylase (TH). Since TH is a catecholaminergic marker which reflects the extent of sympathetic innervation, we evaluated its expression in bone/BM tissues after G-CSF treatment. TH immunofluorescence staining analysis in longitudinal frozen sections of femurs exhibited a strong punctuate staining of the catecholamine-producing enzyme that was increased 10-fold in G-CSF-treated mice compared to control animals. This result was also confirmed by Western blot analysis of equal amount of protein obtained from BM lysates of G-CSF-treated and control animals. Because NE release and removal from the extracellular space determine tissue NE turnover and regulate the sympathetic activation, we evaluated NE release and uptake in sympathetic cervical ganglia (SCG) in organ cultures treated with G-CSF. Although the release of [3H]NE from cultured ganglia was not affected by G-CSF, we found a significant reduction in the reuptake rate. NE clearance from the synaptic cleft is mediated by the Na+-dependent NE transporter (NET). NET is selectively expressed and localized in lipid rafts of the plasma membrane of the noradrenergic nerve terminals and it is internalized as a result of its down-regulation. We thus isolated lipid rafts from BM homogenized in lysis buffer containing 1% Triton X-100 and subjected to sucrose gradient centrifugation. We found that G-CSF induced the redistribution of NET from the plasma membrane as revealed by a strong reduction of NET levels in BM lipid raft-enriched gradient fractions and by an increase of NET levels in BM non-raft fractions following G-CSF treatment. Moreover Q-PCR analysis of catecholamine-inactivating enzymes revealed reduced mRNA levels of the monoamine oxidase (MAO) and the catechol-O-methyltransferase (COMT) enzymes derived from BM of mice injected with G-CSF compared to untreated animals. Altogether, these results demonstrate that G-CSF treatment affects NE turnover impairing the removal of NE from the synaptic cleft and therefore leading to an increase of the concentration of the extracellular NE. A likely consequence is a prolongation of the postsynaptic action of NE on its receptors and ultimately an augmented sympathetic activity in the bone marrow microenvironment.
Functional analysis of the Human Pbx Interacting Protein (HPIP) in Human Hematopoiesis
Authors: Kaur Pawandeep , Stadler Christiane , Ahmed Farid , Cusan Monica , Humphries R. Keith , Hiddemann Wolfgang , Buske Christian , Feuring-Buske Michaela 
Published: April 2020

Abstract
The recognition of novel proteins that regulate human hematopoietic stem cell and early progenitor cell fate is a prime objective in experimental and clinical hematology. Human hematopoietic PBX interacting protein (HPIP), with no significant homology to known proteins, is a 731 amino acid protein, discovered as a novel interacting partner of the PBX homeodomain protein. HPIP has been implicated as a nuclear-cytoplasmic shuttle molecule and shown to have the capacity to bind to the cytoskeleton. It also inhibits the ability of PBX-HOX heterodimers to bind to target sequences and strongly inhibits the transactivation activity of E2A-PBX1 [t(1;19) translocation, which occurs in 25% of pediatric pre-B cell acute lymphocytic leukaemia] (Abramovich C. et al JBC, 2000; Oncogene, 2002). It is highly expressed in human CD34+ progenitor cells, but is silenced in differentiated cells. To gain further insights into the possible functional role of HPIP and its domains and its possible role in a common pathway with HOX transcription co-factor PBX1, HPIP cDNA was cloned in pMSCV-IRES-YFP cassette. Umbilical cord blood enriched with CD34+ population of stem cells was obtained to perform in vitro and in vivo experiments. Mutants, with deletions of the microtubule binding region (ΔMBR-HPIP), and nuclear receptor and PBX1 interacting motif (ΔNRPID-HPIP) were generated and tested in vitro and in vivo. The constitutive expression of HPIP wt and ΔMBR-HPIP in human cord blood cells (CD34+) enhanced erythroid colony formation in CFC assay (p=0.008, n=6) while the ΔNRPID-HPIP mutant nullified the effect. Both mutants of HPIP augmented significantly, the formation of primitive colonies (GEMM and GM) in methylcellulose assay (p≤0.01, n=6) as compared to YFP control and HPIP wt. In replating CFC assays ΔNRPID-HPIP showed an increased number of myeloid colonies (p≤0.01, n=6) and GM (p=ns) colonies but a decrease in granulocytic colonies (p≤0.05, n=6) compared to YFP control and HPIP wt
Identification of Gfi1 as a Direct Target of the Leukemogenic Fusion Protein AML1-ETO9a
Authors: Lo Miao-Chia , Peterson Luke F. , Chen I-Ming , Willman Cheryl L. , Zhang Dong-Er 
Published: April 2020

Abstract
AML1-ETO9a (AE9a), a C-terminal splice variant of t(8;21), is highly leukemogenic in mice. Using combined gene expression and promoter occupancy (ChIP-on-chip) profiling of an AE9a induced murine AML model, we identified Gfi1, a transcriptional repressor important for neutrophil differentiation, as a direct target of AE9a. The level of Gfi1 mRNA was down-regulated following leukemia development. We further show that t(8;21) AML-M2 patient samples have lower Gfi1 mRNA levels compared to non-t(8;21) AML-M2 samples. In addition, Gfi1 protein was down-regulated in the human hematopoietic cell line U937 upon AE or AE9a expression. Two consensus AML1 sites (TGTGGT) located at positions −1750 and +405 from the Gfi1 transcriptional initiation site, respectively, are found in the proximity of the peaks of ChIP-on-chip profiling. We confirmed direct DNA binding of AE9a to these regions by ChIP assays. Furthermore, we showed that the activity of the Gfi1 promoter region −1880 to +481 is repressed by AE and AE9a in luciferase reporter assays. To address the effects of down-regulation of Gfi1 on proliferation, we overexpressed it in a primary AML1-ETO C-terminal mutant (AML1-ETOtr) leukemia cell line. Its overexpression slowed down growth and promoted apoptosis and differentiation of this cell line. In this report, we provide evidence that AE9a directly down-regulates Gfi1, thereby inhibiting hematopoietic differentiation, which may contribute to leukemia development.
Probing the Mitotic History and Developmental Stage of Hematopoietic Cells Using Single Telomere Length Analysis (STELA)
Authors: Hills Mark , Lucke Kai , Eaves Connie J. , Lansdorp Peter M. 
Published: April 2020

Abstract
In most somatic cells, telomeres shorten with each round of cell division. As a result telomere length can be used to assess the mitotic history of cells with some important caveats: the telomere length at birth is highly variable (presumably reflecting different alleles of genes regulating telomere length in the germline), telomere losses can be compensated by telomerase and the overall decline in telomere length includes sporadic, variable losses of telomere repeats resulting from damage to telomeric DNA and/or replication errors. The first caveat can be circumvented by testing cells from the same individual and sporadic telomere losses can be analyzed by single telomere length analysis (STELA). It is more difficult to exclude the effect of telomerase on telomere length but we have previously shown that, despite readily detectable expression of telomerase, hematopoietic stem and progenitor cells show a progressive decline in telomere length with cell division and with age. The clinical relevance of telomere shortening is illustrated in several recent studies linking very short telomeres to bone marrow failure and pulmonary fibrosis. We now show that purified human hematopoietic populations from mobilized peripheral blood (MPB) and cord blood (CB) enriched for stem cells (Lin−CD34+CD38−Rho−) and successively more mature cells display progressively shorter telomeres, pointing to the utility of this method for studies of the mitotic relationship between various stem and progenitor cells. Ultra-short telomeres were readily observed (and found to be significantly more frequent) in terminally differentiated cell populations of MPB, suggesting that sporadic telomere losses occur more frequently during differentiation. When 1000 Lin−CD34+CD38−Rho− cord blood cells were transplanted into two immuno-deficient mice, the most primitive human hematopoietic cells with a CD34+CD38− phenotype lost 3970 and 2790 bp respectively following regeneration in vivo, indicative of ~ 30–80 cell divisions assuming a telomere loss of 50–100 bp/division. Further losses in more differentiated cells were similar to those observed in cells before transplantation. These results illustrate the power of STELA for analysis of telomeres in rare cells and point to a novel strategy to study the turnover and replicative history of cells. Furthermore, these data demonstrate that self-renewal divisions in stem cells rather than additional cell divisions in downstream progenitors are the primary cause of telomere loss following transplantation.
The Dual Bromodomain Protein Brd2 Controls Primary B Cell Mitogenesis and Cell Cycle in Mice Reconstituted with Lentivirus-Transduced HSCs
Authors: Blanton Wanda P. , Wang Fangnian , Liu Hongsheng , Romesser Paul , Faller Douglas , Denis Gerald 
Published: April 2020

Abstract
Transcriptional control of cellular proliferation and differentiation is critically important in hematopoiesis; specifically, the role of chromatin-dependent regulatory processes in this context is poorly understood. The human BRD2 proto-oncogene encodes a double bromodomain protein that binds to acetylated histone H4 in chromatin and is located within the MHC class II locus, suggesting Brd2 plays a role in immunity. However, BRD2 shares no sequence similarity with other MHC genes, nor is Brd2 involved in antigen processing, but rather it plays a role in mitogenic signal transduction. We have previously found that whole-body knockout of Brd2 is lethal to mice. However, when Brd2 was expressed constitutively in the B cells of transgenic mice, Brd2 binds E2F proteins, histone acetylases and Swi/Snf complexes, and co-activates cyclin A leading to B cell lymphoma and leukemia. Importantly, elevated levels of Brd2 have been reported in primary malignant B cells from human and mouse. We therefore hypothesize that Brd2 multiprotein complexes, working through chromatin modification, are crucial in the control of the cell cycle and in the mitogen responsiveness and proliferation of the B cell compartment. To study the effects of Brd2 in B cell development and proliferation, we performed bone marrow transplants of hematopoietic stem cells in a chimeric mouse model. Hematopoietic stem cells were sorted from CD45.1 donor mice with the characteristic ‘side population’ profile by flow cytometry and transduced with lentivirus containing vectors for Brd2 overexpression, shRNA knockdown, or control vectors. Recipient CD45.2 mice were lethally irradiated and a functional immune system was successfully reconstituted with donor cells and CD45.2 competitor BM cells. Mice were immunophenotyped and functional B cell mitogenic capacity was examined by BrdU incorporation into LPS-stimulated B cells. We found that in the spleen, Brd2 expression dramatically expands the CD45.1 (but not CD45.2) B cell compartment at the expense of T cells and renders B cells mitogenically hypersensitive. Compared with control, there was an increase in BrdU incorporation at 24 and 48 hours (29.8% v. 43.5% at T=24 h; 56.9% v. 66.7% at T=48 h). Preliminary results also suggest that B cell development was skewed in the bone marrow and periphery towards B1a phenotype. Moreover, downregulation of Brd2 via shRNA blocked cyclin A transcription and completely arrested B cell development and proliferation. Taken together, these data suggest that Brd2, through epigenetic regulation of the cell cycle, plays an important role in B-lymphopoiesis, proliferation, and stimulation.