bluuebunny/crossref_metadata_2025_split huggingface dataset:
PTPN11 Mutations E76K, D61V, and D61Y Induce Hypersensitivity of Hematopoietic Progenitors to GM-CSF and IL-3.
Authors: Chan Rebecca J. , Li Yanjun , Leedy Melissa B. 
Published: April 2020

Abstract
The childhood leukemia, juvenile myelomonocytic leukemia (JMML), is a lethal disease of young children characterized by spontaneous growth of peripheral blood hematopoietic progenitors and hypersensitivity of hematopoietic progenitors to the cytokine GM-CSF. Notably, hematopoietic progenitors from JMML patients do not typically demonstrate hypersensitivity to the cytokine IL-3. Mutations in the RAS and NF1 genes have long been recognized as pathogenic in this disease, and more recently, mutations in the PTPN11 gene, which encodes Shp-2, a non-receptor protein tyrosine phosphatase, have also been found commonly in leukemic cells from children with JMML. We hypothesized that the mutant Shp-2 molecules observed in JMML patients would induce hypersensitivity of hematopoietic progenitors to GM-CSF. To examine this hypothesis, we subcloned the WT Shp-2 cDNA and three mutant Shp-2 cDNAs found in leukemic cells from children with JMML (E76K, D61V, and D61Y), into the murine stem cell retroviral plasmid pMIEG3 in tandem with EGFP. Each vector was sequenced to verify the desired point mutation and to rule-out unwanted mutations. Murine bone marrow low density mononuclear cells were subjected to fibronectin-assisted retroviral transduction and sorted for EGFP positive cells using fluorescence activated cell sorting. EGFP positive cells were plated into progenitor assays with increasing concentrations of GM-CSF (0, 0.01 0.1, 1, and 10 ng/mL). As predicted, transduction with each of the Shp-2 mutations induced hypersensitivity to GM-CSF compared to vector alone or to WT Shp-2, as evidenced by significantly higher % maximal colony formation at each GM-CSF dose tested (Figure 1).

We next conducted progenitor assays using EGFP positive cells with increasing concentrations of IL-3 (0. 0.01, 0.1, 1, and 10 U/mL). Although hematopoietic progenitors from JMML patients have not been described to be hypersensitive to IL-3, surprisingly, we have observed in preliminary studies that introduction of each of the Shp-2 mutations induced hypersensitivity to IL-3 in a similar fashion to that observed with GM-CSF (Figure 2).

These data demonstrate that somatic PTPN11 mutations observed in children with JMML induce hematopoietic cell growth aberrancies in a manner overlapping with the classical description of JMML, yet also induce unique hematopoietic cell growth characteristics based on the observed hypersensitivity to IL-3. These findings suggest that signaling pathways in addition to the Ras-MAPK cascade may be dysregulated in PTPN11 mutation-bearing hematopoietic cells and provide a novel model for the investigation of new therapeutics in JMML. Current studies are ongoing to examine the effect of these mutations on in vivo hematopoiesis and leukemogenesis.
Oligomerization of Evi-1 Contributes to Recruitment of Transcriptional Corepressor CtBP and Repression of TGF-β Signaling.
Authors: Nitta Eriko , Izutsu Koji , Yamaguchi Yuko , Imai Yoichi , Ogawa Seishi , Chiba Shigeru , Kurokawa Mineo , Hirai Hisamaru 
Published: April 2020

Abstract
Aberrant oligomerization of transcription factors has recently emerged as a prevalent mechanism for activating their oncogenic potential in hematopoietic malignancies. Furthermore, for some chimeric transcription factors generated in leukemia-associated chromosomal translocations including PML-RARα and AML1-ETO, it has also been proposed that aberrant recruitment of corepressor complexes caused by oligomerization is critical for the development of leukemia. Among leukemia-associated transcription factors is Evi-1, which is frequently activated in cases with myeloid malignancies. Evi-1a and Evi-1c, with the latter also called MDS1-Evi-1, are two alternative forms derived from the Evi-1 gene. Evi-1a is a Zn finger transcription factor which recruits CtBP as a transcriptional corepressor and possesses the ability to repress TGF-β signaling. Amino-terminal to the Evi-1a, Evi-1c harbors an additional sequence similar to the PR domain, which was originally identified in PRDI-BF1 and RIZ1. In contrast to frequent upregulation of Evi-1a in myeloid malignancies, expression of Evi-1c widely varies depending on cases. Thus, although several lines of evidence suggest that Evi-1a is involved in leukemic transformation of hematopoietic cells, a role for Evi-1c in leukemogenesis has remained elusive. To elucidate the mechanism that underlies leukemogenesis by Evi-1, we investigated the ability of Evi-1 proteins to form oligomeric complexes. We found that Evi-1a forms a homo-oligomer in mammalian cells, whereas Evi-1c exclusively exists as a monomer. Remarkably, Evi-1c has lost the ability to interact with CtBP and failed to efficiently repress TGF-β signaling. We previously reported that AML1-Evi-1, a chimeric transcription factor generated in t(3;21) leukemia, interacts with CtBP as Evi-1a does. In the current study, we also found that AML1-Evi-1 forms a homo-oligomer, suggesting a close relationship between oligomer formation and CtBP binding. Taken together, these data indicate that homo-oligomerization may contribute to the oncogenic potential of the Evi-1 proteins by regulating the interaction with CtBP. These results also identify a novel function of PR domain to dictate oligomerization of transcription factors.
Effect of G-CSF and Intramyocardial Injection of Bone Marrow Cells on Cardiac Function in Mouse Model of Myocardial Infarction: A Word for Neo-Vasculogenesis.
Authors: Verda Larissa , Luo Kehuan , Han Xiaoqiang , Wasserstrom Andrew , Lomasney Jon , Kowalska Barbara , Burt Richard K. 
Published: April 2020

Abstract
Recent studies suggest that primitive stem cells derived from bone marrow (BM) possess greater functional plasticity that was expected previously. It has been shown that bone marrow stem cells (BMSC) promote repairing mechanisms within myocardium following ischemia/reperfusion models of myocardial infarction (MI). Although it remains unclear whether BMSC transdifferentiate into or just fuse with cardiomyocytes, hemodynamic improvement after intramyocardial BMSC injection as well as after G-CSF injection has been demonstrated. Here, we investigated the contribution of BMSC versus G-CSF administration in myocardial repair following MI. Ten weeks old C57BL/6J mice were irradiated and transplanted with green fluorescent protein (GFP) positive bone marrow cells. Three months later, these mice underwent ligation of left anterior descending branch (LAD) of coronary artery and subsequently divided into three groups. One group (n=7) received G-CSF administration at 200ug/kg for 10 consecutive days. Another group (n=9) was injected with GFP+ marrow cells directly into ischemic heart. The third group was held as control (n=7). One month after coronary ligation we found significant improvement in cardiac function determined as a cardiac output, maximum power and dP/dt, in the G-CSF group compared to control. We evaluated the phenotype of GFP+ cells within myocardium in each treatment group by 488 nm laser-scanning confocal miscroscopy (of whole heart and slides) 35 days after LAD ligation. We found no evidence of myocardial transdifferentiation or cardiomyocyte cell fusion. Instead GFP+ capillaries were present and exclusively located in infarct border zones in both the G-CSF and bone marrow implantation groups, confirmed by anti-factor VIII staining. G-CSF administration and to a lesser extent marrow injection resulted in improved post infarct cardiac function indices. This beneficial effect is not due to transdifferentiation but could be explained by marrow injected or G-CSF mobilized endothelial progenitor cells (EPC) and/or cytokine mediated neo-vasculogenesis.
Evidence That CXCR4+ Neural Tissue-Committed Stem Cells (TCSC) Reside/Hide out in the Bone Marrow and Are Mobilized into the Peripheral Blood during Stroke.
Authors: Kucia Magda , Ping Zhang Y. , Ratajczak Janina , Ildstad Suzanne T. , Shields Chris , Ratajczak Mariusz Z. 
Published: April 2020

Abstract
Several recent studies in animals as well as humans support the notion that bone marrow (BM)-derived cells participate in brain regeneration. However, the identity of the specific cell type responsible for regeneration remains unknown. Recent work from our laboratory revealed that BM contains a highly mobile population of CXCR4+ cells that express mRNA for various markers of early tissue-committed stem cells (TCSC) and which are distinct from hematopoietic stem cells (HSC) (Leukemia2004:18;29–40). In this study we investigated whether BM also contains a mobile pool of TCSC destined to differentiate into neural cells. The TCSC were isolated from bone marrow by employing chemotactic gradient to SDF-1 or by FACS sorting and subsequently evaluated for a presence of early neural markers by i) real time RT-PCR analysis (nestin, GFAP), ii) immunohistochemical staining (nestin, beta-III tubulin), and iii) by employing functional in vitro assays to study ability of these purified cells to form neurospheres. Our data demonstrate that TCSC for neural cells (i) are present in significant amounts in BM harvested from young (1–2 month-old) while being barely detectable in older (1-year-old) mice; ii) reside in populations of murine BM-derived non-adherent non-hematopoietic Sca-1+ CD45− cells and in population of human CXCR4+ CD34+ AC133+ CD45− BMMNC, iii) are mobilized from BM into peripheral blood (PB) during pharmacological mobilization or 24 hours after Bengal-rose induced stroke in mice; iv) SDF-1 is highly upregulated in damaged brain tissue, and v) TCSC are chemoattracted for potential brain regeneration in SDF-1-CXCR4, dependent manner. Thus, we conclude that bone marrow is a potential source of TCSC for brain repair and since purified CD45+ HSC neither express neuronal markers nor differentiate in vitro into neurospheres, we provide for a first time evidence that neural TCSC residing in bone marrow but not “plastic” HSC account for neural differentiation of BM-derived cells. Furthermore, our observation that the number of marrow derived mobile/circulating neural TCSC is the highest in BM of young animals and decreases with age provides a novel insight into aging and may explain why the brain regeneration process becomes less effective in older individuals. Finally, these observations provide rationale for further studies aimed at optimizing therapeutic brain regeneration by BM-derived neural TCSC.
Rapid Risk-Profiling Including Fast Donor Search within a Multi-Center Trial Allows for Early Allogeneic Stem Cell Transplantation during Aplasia in Patients with High-Risk Acute Myeloid Leukemia.
Authors: Schaich Markus Andreas , Fuessel Monika , Bornhaeuser Martin , Thiede Christian , Mohr Brigitte , Illmer Thomas , Schaekel Ulrike , Prange-Krex Gabriele , Oelschlaegel Uta , Ehninger Gerhard ,   on behalf of the DSIL AML2003 study group
Published: April 2020

Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) remains the only curative treatment option for most high-risk patients with acute myeloid leukemia (AML). However, many high-risk patients regenerate with blasts or relapse early after induction therapy. Thus, consolidation with allogeneic HSCT in first CR is often not possible. Performing allogeneic HSCT within induction therapy can circumvent these problems and may furthermore reduce cumulative toxicity in high risk patients.
Therefore, the prospective randomized treatment trial AML2003 for patients &lt;= 60 years was set up, to investigate the feasibility and value of an intensified treatment strategy, i.e. early allogeneic stem cell transplantation in aplasia after induction therapy, for high risk AML patients in a multi-center setting. To achieve this goal, rapid analysis of cytogenetics, FLT3 status and HLA-types of the patient and possible family donors is of utmost importance. This fast search diagnostics together with routine analyses of morphology and immunophenotyping is accomplished centrally in all enclosed patients. Furthermore, in all patients the likelihood to find an unrelated donor is checked by an internet search in the BMDW database.
Within the first 8 months 107 AML patients with a median age of 48 (17–60) years were included in the study. Fast search diagnostics was complete within a median of 15 (range 5–31) days after arrival of the bone marrow samples for all patients. 57/107 patients were randomized into the intensified treatment arms. Out of these 25 (44%) patients with high risk characteristics have been identified. A suitable related or unrelated donor was found for 22 (88%) of those high-risk patients. Nine of those high risk patients with a donor (41%) received early allogeneic stem cell transplantation in aplasia after the first (n=4) or the second (n=5) induction therapy course on an intend to treat basis within the protocol. Three were transplanted with stem cells of related and six of unrelated donors. The preparative regimen consisted of melphalan 150mg/m2 and fludarabine 150mg/m2. So far no treatment associated death had to be recognized.
These encouraging preliminary results show that fast risk-profiling and early donor-search is feasible in a large multi-center study. This leads to a significant proportion of early allogeneic stem cell transplants in aplasia after induction therapy within the group of high risk AML patients, which may improve the disastrous prognosis of this group of patients in the future.
Identification of the Genomic Breakpoints at 1p36.3 and 3q21.3 in Five MDS/AML(M4) Patients with t(1;3)(p36;q21) Translocation. GATA2 Mutation May Be Involved in Leukemogenesis.
Authors: Xinh Phan Thi , Vu Hoang Anh , Taketazu Fumitoshi , Fujisawa Shin , Tanaka Hideo , Yagasaki Fumiharu , Toyoda Atsushi , Sakaki Yoshiyuki , Rowley Janet D. , Tokunaga Katsushi , Sato Yuko 
Published: April 2020

Abstract
Reciprocal translocation t(1;3)(p36;q21) is found in a subset of MDS/AML characterized by trilineage dysplasia, especially dysmegakaryopoiesis and a poor prognosis. In this leukemia, ectopic expression of MEL1 at 1p36.3 driven by the promoter of RPN1 at 3q21 is thought to be associated with the pathogenesis. Recently, we reported the identification of 2 kinds of the 1p36 breakpoint (BP) region in 3 patients with t(1;3), 1) a 70.7-kb region within the first intron of MEL1 in one patient, and 2) a 29.2-kb region in the 5′ region of MEL1 in the remaining two. However, the precise BP at 3q21 has not yet been determined. Here, we report that we have defined the 3q21 BP in 4 patients, and the 1p36.3 BP in 2 additional patients with FISH using bone marrow cells obtained from 4 patients (4 with MDS-related AML-M4 and one with CMML). To identify the BPs and deletions at 1p36.3 or at 3q21.3, we used 18 non-contig BAC/PAC probes and 7 contig BAC/PAC probes, respectively. FISH study revealed that the 3q21 BPs were clustered within RP11-475N22 probe in the 4 patients, in the 5′ region of GATA2. The 1p36.3 BP was found within the first intron of MEL1 in one patient, but in a 68.5-kb region which included a part of the first intron and a region of 5′ site of MEL1 in the other. Therefore, 3 BP regions at 1p36 were found among the 5 patients. No deletions were detected in the vicinity of the 1p36 BP or 3q21 BP in any of our 5 patients, although Lahortiga et al., detected a large deletion at the both BPs in their patients with MDS (RAEB) and t(1;3) (Oncogene 23:311–316, 2004). In Patient 1 (the 1p36 BP was in the first intron of MEL1) and Patient 3 (the 1p36 BP was in 5′ site of MEL1), the expression patterns of MEL1, ARPM2, and GATA2 which are located near the BPs were analyzed with RT-PCR. ARPM2 was not overexpressed in the two patients, while MEL1 was overexpressed in Patient 1, but not in Patient 3 when the expression level was compared with that of normal bone marrow cells. It is reported that GATA2, one of the zinc finger transcription factors, serves as a regulator of genes controlling proliferative capacity of early hematopoietic cells, and that two zinc finger domains of GATA2 are closely involved in inducing megakaryocytic differentiation. Interestingly, a deleted GATA2 transcript was detected in Patient 3 together with a wild type GATA2 transcript. The short GATA2 transcript contained 1009 nucleotides deletion from exon 3 to exon 6 with a frameshift, resulting in lacking the two zinc finger domains. Thus, there is the possibility that dysmegakaryopoiesis in this patient is associated with the expression of GATA2 without the zinc finger domains.
Improved Heart Function after the Combined Treatment of Patients Undergoing Coronary Artery Bypass Grafting through Stem Cell Therapy and Transmyocardial Laser Revascularisation.
Authors: Stockschlaeder Marcus , Ghodsizad Ali , Gams Emmeran , Poll L. , Stoldt Volker , Piechaczek E. R. , Feifel N. , Klein Michael , Scharf Ruediger E. 
Published: April 2020

Abstract
To improve tissue regeneration of ischemic myocardium, autologous bone marrow-derived stem cells have been injected intramyocardially in five patients undergoing coronary artery bypass grafting and transmyocardial laser revascularization. We have established an innovative method for the intraoperative isolation of CD133+-stem cells in less than 3 hours. After induction of general anesthesia, approx. 60 – 240 ml of bone marrow were harvested from the posterior iliac crest and processed in the operating room under GMP conditions using the automated cell selection device CliniMACS. Following standard coronary artery bypass grafting, laser channels were shot in predefined areas within the hibernating myocardium. Subsequently, autologous CD133+-stem cells (1.9–9.7 x 106 cells; purity up to 97%) were injected in a predefined pattern around the laser channels (0.5–1.0x106 cells/channel). Three months postoperatively, significant improvement of cardiac function as assessed by transthoracal echocardiography (ejection fraction before treatment: 25–30% - after treatment: 38–46%; left ventricular enddiastolic volume before treatment: 58 – 64 mm - after treatment: 40–44 mm; endsystolic wall thickness before treatment: 5–7 mm - after treatment 9–13 mm) and improvement of cardiac wall motion (electrocardiographically-triggered magnetic resonance imaging) were documented. This significant gain in heart function might be partly explained through the synergistic angiogenic effect of transmyocardial laser revascularization leading to a localized inflammatory response and the cellular effects of intramyocardially injected pluripotent bone marrow-derived stem cells promoting tissue repair. Through the intraoperative isolation of CD133+-cells, this effective treatment of ischemic myocardium can be applied to patients scheduled both for elective and for emergency revascularisation procedures.
Validation Study on the Prediction of Response to Imatinib Mesylate in Chronic Myeloid Leukemia (CML) Patients by Genome-Wide cDNA Microarray Analysis.
Authors: Kagami Yoshitoyo , Katagiri Toyomasa , Kaneta Yasuyuki , Tsuruo Takasi , Nakamura Yusuke , Ohno Ryuzou 
Published: April 2020

Abstract
Imatinib mesylate (IM) is highly effective in the first line therapy of CML. However, it remains unknown whether IM cures CML or not, and other therapeutic modalities like stem cell transplantation sometimes become necessary. Predicting the therapeutic effect of IM before the start of therapy would be useful, because other kinds of therapy could be planed during IM treatment. Previously, in order to predict the effect of IM therapy in CML, we established a prediction system by cDNA microarray containing 23,040 genes. According to the profiling data of RNA expression of peripheral blood mononuclear cells in 22 CML patients, who had previously failed in interferon-α therapy, 79 genes were identified that were expressed differentially between responders and non-responders to IM. By a prediction scoring system, 15 out of the 79 genes were selected which discriminated the two populations most clearly.(Kaneta Y et al. Jpn J Cancer Res 93, 849–856,2002) This time we applied this scoring system to another cohort of chronic or accerelated phase of CML patients who would receive IM as monotherapy. So far, 42 patients were analyzed. Of 40 patients who attained major cytogenetic response (MCR) by 6 months from the start of therapy, 38 (95%) had been predicted as the responder by the prediction system. On the other hand, two patients whose effect was less than MCR had been predicted as the non-responder. These data indicate that our prediction system for IM using cDNA microarray could effectively discriminate responders from non-responders before the start of therapy.
The Antileukemic Effect of NK and NK-T Cells Against Chronic Myelogenous Leukemia Is Dominated by Interactions between Activating Molecules and Their Ligands Which Overcome Inhibitory KIR-MHC Class I Interaction.
Authors: Sconocchia Giuseppe , Lau Michelle , Provenzano Maurizio , Rezvani Katayoun , Fujiwara Hiroshi , Ferrone Soldano , Barrett John 
Published: April 2020

Abstract
We compared the interactions of autologous, HLA-identical and mismatched CD56+ cells with chronic myelogenous leukemia (CML) CD34+ cells. In 14 donor-recipient pairs, HLA-matched donor CD56+ cells inhibited CML CFU-GM at an E/T ratio of 10:1 (mean inhibition 42±9%). This was comparable to the 39.5±7% inhibition observed with 14 HLA-mismatched unrelated CD56+ cell donors, indicating that KIR incompatibility was not required for the antileukemic effect. Both CD56+CD3- (NK) and CD56+CD3+(NK-T) subsets inhibited colony formation of CD34+ CML cells but not normal CD34+ cells from the stem cell donor, suggesting that CD56+ cells recognized surface molecules specific to CML cells. Using indirect immunofluorescence to detect NKG2D ligands, 11/12 CML CD34+ samples were found to be MICA/B+ positive, while normal CD34 cells were uniformly negative. Five of six MICA/B+ CML samples bound a NKG2D/FC+ chimeric molecule indicating the presence of functional ligands. CML CFU-GM inhibition was partially blocked by anti-NKG2D suggesting that NKG2D-MICA/B interactions contributed to the antileukemic activity of NK and NK-T cells. Colony formation of MICA/B+ CML cells was also inhibited by caspase-blocked HLA matched donor CD56+ NK cells indicating that the effect was not FAS-dependent. However, soluble MICA/B was detected in patient serum by immunoprecipitation and CML patient serum blocked anti-MICA/B binding to CML cells, suggesting that in vivo anti-leukemic activity of CD56+ cells is diminished. Furthermore NKG2D expression on CD3+ cells was found to be low in 3 of 4 CML patients, which was accompanied by a reduced IL-10 production upon NKG2D ligation by an anti-NKG2D moAb. These studies confirm early observations of KIR-independent antileukemic cytotoxicity by autologous and HLA matched NK cells, implicating pro-cytotoxic signaling via NKG2D and its ligand MICA/B. They also reveal mechanisms whereby CML cells may escape from NK regulation in vivo.