bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD73-Independent Toxicity of Fludarabine on Adult Normal Bone Marrow Mesenchymal Cells (NBMMC).
Authors: Pebrel-Richard Celine , Veyrat-Masson Richard , Boiret-Dupré Nathalie , Rapatel Chantal , Berger Juliette , Tournilhac Olivier , Boisgard Stephane , Chassagne Jacques , Berger Marc G. 
Published: April 2020

Abstract
The purine analogue fludarabine is efficient in treating advanced, chronic B-cell lymphocytic leukemia (CLL). We and others have shown that front-line fludarabine treatment impaired the subsequent mobilization of hematopoietic CD34+ cells by G-CSF. This might imply an effect on hematopoietic and/or mesenchymal compartments. We also previously showed that native mesenchymal cells (MC), like expanded MC, express the 5′-ecto-nucleotidase (CD73) which should aid the entry of fludarabine into the cell. So we investigated the toxicity of fludarabine on NBMMC.
We incubated expanded (P1) and fresh NBMMC from femoral heads with increasing doses (0.1 to 6 mMol) of fludarabine for 3 days, and assessed its effect on cell survival (annexin V staining), CFU-F frequency, cell proliferation and immunophenotyping. We used α,β, methylene-adenosine 5 diphosphate (MADP) to inhibit CD73 in order to evaluate its role in the trans-membrane passage of fludarabine.
We observed a direct, dose-dependent toxicity of fludarabine on expanded mesenchymal cells, after 3 days incubation, characterized by apoptosis of 48% of MC. The mesenchymal progenitor cells were more sensitive; their frequency amongst the viable cells, being decreased about 5-fold from the 1mMol dose, to 13-fold at the highest dose, where there were far fewer colonies containing&gt;50 cells. Even after fludarabine removal, viable MC showed an impaired ability to proliferate at the next passage. Fresh NBMMC, incubated under same conditions, showed a similar but lower sensitivity than did expanded (P1) mesenchymal cells, as indicated by a shift in the dose-response curve to a 5-fold decrease at the highest dose. Again, we observed a decreasing proportion of large colonies. MADP provided no significant protection against the effect of fludarabine on fresh or expanded cells. The phenotype of progeny was comparable to that of the controls.
In conclusion, we have shown for the first time that adult NBMMC are sensitive to fludarabine and could prove to be an unexpected in vivo target. The differences between un-manipulated and expanded cells suggest a biologic alteration during culture. The lack of protective effect of MADP and the absence of lower expression of CD73 on resistant cells suggest another route of trans-membrane passage for fludarabine. The consequences of low dose fludarabine incubation on MC properties are currently being studied.
Usefulness of Wharton’s Jelly, Cord Blood, and Adipose Tissue as Alternative Sources of Mesenchymal Stem Cells.
Authors: Jang Jun Ho , Lee Hyun Woo , Eom Young-Woo , Kang Seok Yun , Park Joon Seong , Choi Jin Hyuk , Kim Hugh C. 
Published: April 2020

Abstract
Mesenchymal stem cells (MSCs) are a highly promising source of adult stem cells for purposes of cell therapy and tissue repair in the field of regenerative medicine. Although the most studied and accessible source of MSC is the bone marrow, the clinical use of bone marrow-derived MSCs (BMSCs) has presented problems, including pain, morbidity, and low cell number upon harvest. For those reasons, we isolated, cultured, and characterized MSCs from a number of tissues; including wharton’s jelly, cord blood, and adipose tissues that were discarded routinely in the past, and evaluated the usefulness of these MSCs compared to BMSCs. Proliferation ability of Wharton’s jelly-derived MSCs (WJ-MSCs), Cord blood-derived MSCs (CB-MSCs), or adipose tissue-derived MSCs (ASCs) was lost at passage 8–10 (22–27 population doubling), passage 7–10, or passage 7–12 (45–50 population doubling), respectively. WJ-MSCs, CB-MSCs, and ASCs expressed CD73, CD90, and CD105, CD90, CD105, and CD166, and CD44, CD73, CD90, and CD166, respectively, were absent for CD14, CD31, and CD45, and differentiated into osteoblast, adipocyte, and chondrogenic lineages under appropriate culture condition. In this study, like BMSCs, WJ-MSCs, CB-MSCs, and ASCs expressed similar cell surface antigens, were able to differentiate into mesenchymal lineages, and possessed highly proliferation potential. Therefore, MSCs isolated from wharton’s jelly, cord blood, and adipose tissue may become useful alternative sources of MSCs to cell therapy and tissue repair in the field of regenerative medicine.
Characterization of a Bone Marrow Derived Mesenchymal Cell Line from Nude Rat Assembling Tumor Stem/Progenitor Cell Properties In Vitro and In Vivo.
Authors: Wang Mengyu , Ju Jingfang , Saeboe-Larssen Stein , Mikalsen Svein-Ole , Olsen Elena , Nesland Jahn M. , Fodstad Oystein , Maelandsmo Gunhild , Kvalheim Gunnar 
Published: April 2020

Abstract
Here we report the in vitro and in vivo properties of a spontaneously transformed nude rat mesenchymal stem cells (rTMSCs) following culture passages 23. When this cell line was compared with early passages of mesenchymal cells (rMSCs) from the same animal, no apparent differences could be seen with regard to the phenotype and Gap junctions’ connexin 43. However, the telomerase activity in rTMSCs is lower than in the rMSCs. Karyotyping of rTMSC gave a trisome 6 phenotype. Microarry analysis of rTMSC shows over expression of the transforming growth factor family including TGF-beta and BMp6. Both rMSC and rTMSC had equal capability to differentiate into adipocytic and “neural like” linage. In contrast rTMSC was unable to differentiate into bone and only this cell line form spheroids in cultures.
Subcutaneously injection of rTMSC in doses down to 2x 104 into nude/nude mice formed tumor in all animals tested. Examination of the tumor assembles morphology like an immature sarcoma. When 100–200 SP + cells were administered to the animals, tumor could also be formed. After intravenous injections and direct injections into left heart ventricle all animals developed metastasis in the lung; abdominal cavity; bone and skin. The rTMSC has been stabled marked with reporter gene GFP and Leucipherase. Currently the proliferation and migration capacity of the linage-differentiated rTMSC is studied in vivo by imaging and these results will be presented. We conclude that our normal and transformed rMSCs could be potential useful for further biological studies on mesenchymal cells both in vitro and in vivo.
Active Oral Regimen for Elderly Adults with Newly Diagnosed Acute Myelogenous Leukemia (AML): Phase I Trial of Oral Tipifarnib (T) Combined with Oral Etoposide (E) for Adults ≥Age 70 Who Are Not Candidates for Traditional Cytotoxic Chemotherapy (TCC).
Authors: Karp Judith E. , Feldman Eric J. , Morris Lawrence , Greer Jacqueline , Ironside Valerie , Smith B. Douglas , Ritchie Ellen , Gore Steven D. , Levis Mark J. , Kaufmann Scott H. , Malek Sami , Wright John J. 
Published: April 2020

Abstract
T is a Farnesyltransferase inhibitor with clinical activity in elderly adults with newly diagnosed AML who are not candidates for TCC. For adults ≥ age 65 with poor-risk AML, T 600 mg BID for 21 out of every 28–63 days induces complete remissions (CR) in 15%, median duration 7.3 mos, median survival 18 mos. In vitro, T+E exhibits synergistic anti-AML effects. In an attempt to increase CR rates in elderly AML pts, we are conducting a Phase I trial of oral T+E, with escalating doses of both drugs and 14 vs. 21 days of T every 28–63 days (median time to beginning cycle 2 is Day 31, range 29–43). To date, 60 adults ≥ age 70 (median 77, range 71–91) have been treated at 10 dose levels of T (300, 400 or 600 mg BID, 14 or 21 days) + E (100, 150 or 200 mg daily days 1–3 and 8–10) and have received 145 cycles (median 3, range 1–7). A total of 35 (58%) have had secondary AML and 31 (52%) have had adverse cytogenetics. Marrow blast percent was median 55% (range 20–96%), with 75% &gt;30,000 blasts. Nonhematologic toxicities include oropharyngeal mucositis (15% ≥ grade 2), CNS (overall 30%, with 80% of those ≤ grade 2), renal (6%), hyperbilirubinemia (6%), rash (20%) and fatigue (20%). Toxicities occurred across all T+E dose levels tested to date. Documented infections occurred in 16 (27%) pts during cycle 1 and 22/145 (14%) cycles. Twenty six pts (43%) required hospitalization during cycle 1 for median 7 days (range 2–28). Overall hospitalization rate for all cycles was 28% (41/145). Ten pts (17%) died on study: 6 during cycle 1 (4 infection, 1 emphysema, 1 cerebrovascular) and 4 during cycles 2–4 (3 acute myocardial infarctions, 1 ventricular arrhythmia). For 56 evaluable pts, 12 (21%) have achieved CR and 11(20%) have achieved partial remission/hematologic improvement (PR/HI). All pts achieving CR have done so within 2 cycles (9/12 after cycle 1). Eight of 30 pts (27%) receiving T ≥ 400 mg BID have achieved CR. Median age of CR pts is 79 (range 71–86), 8 (67%) have secondary AML, and 5 (42%) have complex cytogenetics. CR durations to date range from 2+ to 13.5+ mos, with median 7+ months. Three of 12 have relapsed at 3, 10, and 11 mos, 2 have been retreated with T+E, and both have achieved 2nd CRs. In sum, the oral regimen of T+E is tolerable and feasible on an outpatient basis for elderly adults with newly diagnosed AML who are not candidates for TCC, with the suggestion of improvement of CR rates over T alone.
Src Kinase Inhibitors Enhance ATRA-Mediated Differentiation of Myeloid Cells.
Authors: Miranda Michelle B. , Johnson Daniel E. 
Published: April 2020

Abstract
Myeloid leukemias are characterized by a blockade in differentiation resulting in accumulation of proliferating progenitor cells and a lack of mature granulocytes and monocytes. In acute promyelocytic leukemia (APL), a subtype of AML, treatment with all-trans retinoic acid (ATRA) has been shown to overcome this blockade in differentiation and is the current standard of care for APL patients. However, other forms of myeloid leukemia show no response to ATRA. Src family kinases (SFK) have been shown to be activated during normal myeloid development by both proliferation-inducing cytokines, such as IL-3, and differentiation-promoting factors, such as G-CSF. To elucidate the role of src family kinases during myeloid differentiation, we examined the impact of SFK inhibitors, PP1 and PP2 on ATRA-mediated differentiation of myeloid cell lines. Interestingly, PP1 potentiated ATRA-mediated myeloid differentiation of HL-60 cells, inducing a two-fold increase in differentiated cells at 72 hours as determined by CD11b staining, NBT reduction and morphological analysis. To determine whether enhancement of ATRA-mediated differentiation was specific for PP1, or could be promoted by a different src family kinase inhibitor, we evaluated the impact of PP2. PP2 was found to exhibit similar effects on ATRA-induced myeloid differentiation of HL-60 cells. Additionally, both PP1 and PP2 enhanced ATRA-induced monocytic differentiation of U937 cells and granulocytic differentiation of NB-4 cells. Interestingly, PP1 was found to enhance ATRA-induced differentiation even when added 24 hours after addition of ATRA. The impact of PP1 on ATRA-mediated myeloid differentiation was dependent on signaling via the MEK/ERK pathway since pre-treatment with U0126, a MEK-1/-2 inhibitor, attenuated myeloid differentiation induced by the combination of RA and PP1. Reporter assays utilizing an RARE-luciferase construct indicate that PP1 and PP2 do not enhance ATRA-mediated differentiation by promoting the transcriptional activity of the retinoic acid receptor (RAR). Together, our results demonstrate that PP1 and PP2 potently enhance ATRA-induced myeloid differentiation of AML cell lines and suggest that src inhibition in combination with ATRA may be a useful treatment for AML. Studies evaluating the impact of src inhibition on ATRA-mediated differentiation of primary AML cells are currently underway.
Tumor Necrosis Factor-Mediated Inflammation Exacerbates Genomic Instability and Promotes Leukemia Development.
Authors: Li June , Sejas Daniel P. , Zhang Xiaoling , Rani Reena , Pang Qishen 
Published: April 2020

Abstract
A correlation has been shown between elevated circulating pro-inflammatory cytokines and anemia in patients with leukemia-related diseases but direct evidence for the mechanistic link between inflammation and leukemia is lacking. We have investigated the role of the pro-inflammatory cytokine tumor necrosis factor a (TNF-a) in leukemic development using the disease model of Fanconi anemia, a genetic disorder clinically characterized by congenital anomalies, progressive bone marrow failure, and a high risk of developing leukemia and other cancers. We demonstrate that long-term TNF-a exposure of Fanconi bone marrow progenitors enhances inflammatory response, promotes clonal proliferation, and ultimately leads to leukemia development. NF-kB activation is required for TNF-a-promoted progenitor growth and early stage of leukemia development but is dispensable for the maintenance of leukemic transformation. Pharmacological elimination of TNF-a-induced reactive oxygen species reduces inflammation and delays leukemia development, suggesting that oxidative damage may serve as a link between inflammation and leukemia. In addition, TNF-a-promoted leukemic cells show persistent DNA damage response and increased genomic instability. Furthermore, correction of Fanconi genetic deficiency prevents clonal progenitor proliferation and leukemic transformation by eliminating oxidative DNA damage. Thus, inflammation can exacerbate genomic instability and contribute to leukemia development. This may explain, at least in part, why cells with genomic instability have high predisposition to cancer. Our study underscores therapeutic value of anti-oxidants and anti-inflammation towards tumorigenesis.
Expression of CD271 in Normal and Malignant Hematopoietic Progenitor and Stem Cells.
Authors: Jiang Ying , Powers Jennifer , Muschler George F. , Maciejewski Jaroslaw P. 
Published: April 2020

Abstract
The relationship between mesenchymal (MSC) and hematopoietic stem cells (HSC) involves close interaction in the marrow microenvironment. They also may share certain phenotypic features and even possibly a multipotent common precursor. CD271 (also known as LNGFR, p75NTR) has been reported to be a suitable marker of resting primitive MSCs. CD271 and its ligands, neurotrophins (NTs, e.g. nerve growth factor) also play important roles in hematopoiesis, such as enhancement of colony formation by human bone marrow progenitor cells. Dysregulated expression or mutations of NT receptors were associated with the initiation and progression of various malignancies. Based on expression of CD271 in the mesenchymal stem cell compartment, we have hypothesized that this antigen may also be present on a specific subset of normal or malignant hematopoietic progenitor and stem cells. First, we investigated the expression and function of CD271 in normal bone marrow. In controls (n=11), expression of surface CD271 was present on 1.67 ± 1.26% and 2.78 ± 4.50% of CD45+ and CD45− cell populations, respectively. Within the immature hematopoietic compartment, CD271 was expressed on 2.16 ± 1.16% cells defined by CD45 and CD34 expression. When the properties of CD271-expressing CD34 cells were studied in methylcellulose colony assays, those sorted for CD271 antigen showed a much lower clonogenic potential as compared to the CD271-CD34+ cell fractions (10.5 ± 2.12 vs. 183.5 ± 17.68). Subsequently, we investigated whether leukemic cell lines express CD271. We have analyzed HL60, U937, UT7, KG1, K562, HEL, MKN95, NB4 and Kasumi cells using flow cytometry. CD271 was found to be strongly expressed (72.8%) on KG1 AML. Smaller but clearly distinct subpopulations of HEL cells (7.8%) and NB4 cells (5.2%, APL cells) expressed CD271 suggesting either aberrant differentiation in some subpopulations of these cells or the presence of an early precursor population. We then assessed whether CD271 is expressed by CD34+ blasts in patients with myelodysplastic syndrome (MDS n=9) and AML (n=9). CD271 antigen was found on blast cells in 5/9 MDS (1 CMML, 1 RARS, 2 MDS-derived sAML, 1 RA) patients (7.16 ± 1.97% of CD271+ cells within blasts) and 3/9 AML patients (9.70 ± 3.65% of CD271+ cells within blast gate) suggesting that this fraction may differ functionally from the remaining majority of blasts. This is the first report that relates CD271 expression to MDS. The properties of a distinct CD34+ cell population expressing CD271 in myeloid malignancies are currently under investigation. Results of these studies may point towards the presence of a specific subset of precursor/stem cells with clinical and prognostic implications.
Comparison of Differential microRNA Expression Profiles between CD34+CD38− and CD34+CD38+ Cells in Both Umbilical Cord Blood and Acute Myeloid Leukemia.
Authors: Li Xin , Zhang Jiahua , Liu Jun , Huang Shiang 
Published: April 2020

Abstract
Recent studies demonstrated that some miRNAs played an important role in human normal and malignant hematopoiesis. MicroRNA expression is tissue and developmental stage restricted. Therefore, in order to address the roles of miRNAs in the regulation and maintenance of normal hematopoietiesis and leukemogenesis, in this study, we performed an analysis of differential genome-wide miRNA expressions of CD34+CD38− and CD34+CD38+ cells from human umbilical cord blood(CB) and acute myeloid leukemia(AML) bone marrow, respectively.
Methods Mononuclear cells from cord blood (CB) and AML bone marrow were stained with monoclonal antibodies. CD34+CD38− and CD34+CD38+ cells were sorted using FACSVantageSE. The mRNA were extracted and hybridized to miRCUURY array chip (Exiqon). The data were analyzed with GeneSpring and informatics technique. Real-time PCR for several miRNAs were performed to confirm the results obtained by miRNA chip analysis.
Results Based on the criteria of two or more fold changes of miRNA expression level, 73 miRNA overexpressions and 14 miRNA downexpressions were observed within CD34+CD38+ cells in comparison with CD34+CD38− cells in CB. Interestingly, only one overexpression and10 downexpressions of miRNAs were found within CD34+CD38+ cells as compared with CD34+CD38− cells in AML. This might suggest the less differential and maturational activities during the transition from CD34+CD38− cells to CD34+CD38+ cells in AML as compared with their normal counterpart. In comparison between CB and AML, 94 over-expressed and 28 down-expressed miRNAs were displayed in AML CD34+CD38− cells. More specifically, miR-155, -190 were down-expressed in both CB and leukemic CD34+CD38+ subsets, while miR-220, -499 were over-expressed in CB CD34+CD38+ cells and down-expressed in leukemic CD34+CD38+ cells. Putative hematopoiesis-specific target genes were analyzed with informatics technique. We found that each miRNA had several hundred genes as putative targets and each target gene had more than one miRNA predicted to be its regulator. For example, hsa-miR-509 has 561 putative target genes, of which IL1A, IL3RA1, FGFR2, SMAD2 are hematopoiesis-specific, and SMAD2 has 34 miRNAs as its regulators.
Conclusion: The miRNAs play important roles in the developments of both hematopoiesis and leukemogenesis. The analysis of differential microRNA expression profiles may be a powerful tool to allow us insight on the mechanisms of regulations of hematopoietic stem cells and leukemic stem cells.
Establishment of a NOD/SCID/γc−/−-Dependent Lymphoid Leukemia Cell Line, D593, with Novel RUNX1-LAF4 Fusion Gene.
Authors: Abe Akihiro , Ninomiya Manabu , Imagama Shizuka , Suzuki Momoko , Hayakawa Fumihiko , Katsumi Akira , Kiyoi Hitoshi , Kojima Seiji , Naoe Tomoki 
Published: April 2020

Abstract
We established a NOD/SCID/γc−/−(NOG mouse)-dependent human lymphoid leukemia cell line, D593, by repeated xenotransplantation of pediatric T-cell acute lymphoblastic leukemia cells with the translocation t(2;21). The cell line, D-593, could be serially transplanted from mouse to mouse over a 2-year period. D593 had the same immuno-phenotype as the original leukemia cells: positive for CD2, 5, 7, 14, and 34, and negative for CD3, 4, 8, 19, and 41a. Cytoplasmic CD3 was positive and the rearrangement of T-cell receptor was detected by Southern blot analysis. A previously unreported translocation of t(2;21)(q11;q22) was observed in both the original patient sample and D593. The split signal of RUNX1 was detected by fluorescence in site hybridization in D593 indicating the involvement of RUNX1. Using 3′-RACE and RT-PCR analysis, we identified novel chimeric transcripts of RUNX1-LAF4 joining exon 7 of RUNX1 to exon 4 of LAF4. In the transplanted NOG mice, D593 homed into the trabecular endosteal region of bone marrow (BM), and proliferated from the endosteum to medulla. At the late stage of engraftment, the BM was filled with human lymphoblasts and metastases into the trabecular of the spleen and Glisson’s sheath of the liver were also observed. These findings suggest that D593 is a useful cell line to study not only the leukemia-related biology of RUNX1-LAF4 but also the novel therapeutic model against core-binding factor (CBF) leukemia.
Inv(11)(q21q23) Fuses MLL to the NOTCH Co-Activator Mastermind-Like 2 in Secondary T Cell Acute Lymphoblastic Leukemia.
Authors: Metzler Markus , Zuna Jan , Staege Martin S. , Harder Lana , Meyer Claus , Flohr Thomas , Meerpohl Joerg , Fronkova Eva , Langer Thorsten , Harbott Jochen , Trka Jan , Siebert Reiner , Marschalek Rolf , Niemeyer Charlotte Marie , Rascher Wolfgang 
Published: April 2020

Abstract
Development of a secondary leukemia after chemotherapeutic treatment for childhood acute leukemia is associated with rearrangements of the MLL gene on chromosome 11q23 and characteristically results in acute myeloid leukemia (AML). We identified the NOTCH co-activator Mastermind-like 2 as MLL fusion partner in two cases of pediatric secondary leukemia. The MLL-MAML2 fusion results from a cryptic inv(11)(q21q23) only detectable by interphase fluorescence in situ hybridization with an MLL split signal probe. With a latency of twenty months and five years after initial diagnosis for MLL negative AML and precursor B acute lymphoblastic leukemia (ALL), respectively, both patients developed a therapy-related T-cell acute lymphoblastic leukemia.
MAML2 is the first MLL fusion partner involved in human Notch-signaling and was only recently identified as recurrent translocation fusion partner in a subset of salivary gland tumors. The genomic MLL breakpoint shows similar localization and sequence features described for etoposide induced treatment-related AML. MLL-MAML2 positive cells were detectable up to two years prior to clinical apparent secondary leukemia in one case. The discrepant dynamics of clone expansion quantified by either the genomic fusion sequence or Ig/TCR gene rearrangements as clone specific markers suggests a selection process within the inversion positive population and the need for additional mutation events to promote overt leukemic disease. Whole genome expression profiles demonstrated differential expression of both typical MLL and NOTCH downstream genes, which suggests a modulatory role of the MAML2 transcriptional activation domain in MLL leukemogenesis and lineage assignment induced by the MLL-MAML2 fusion protein.
MSM21620813
WSF3401600.