bluuebunny/crossref_metadata_2025_split huggingface dataset:
Leukemic CD34+CD38- Cells Display Conserved Differential Expression of Many ATP Binding-Cassette Transporter Genes.
Authors: Raaijmakers Marc H.G.P. , de Grouw Elke P.L.M. , van de Locht Louis T.F. , van der Reijden Bert A. , de Witte Theo J.M. , Jansen Joop H. , Raymakers Reinier A.P. 
Published: April 2020

Abstract
In most cases of acute myeloid leukemia (AML) CD34+CD38− cells are considered to be stem cells, responsible for the maintenance and relapse of AML. ATP binding cassette transporters function in the extrusion of xenobiotics and chemotherapeutical compounds, and may be involved in therapy resistance. Elucidation of mechanisms conferring drug resistance to CD34+CD38− cells is essential to provide novel targets for stem cell eradication in AML. We studied gene expression of all 45 transmembrane ABC transporters (the complete ABCA, B, C, D and G family) in human hematopoietic CD34+CD38− cells and more committed CD34+CD38+ progenitor cells, from healthy donors and patients with non-hematological diseases (N=11) and AML patients (N=11). Gene expression was assessed using a novel real-time RT-PCR approach with micro fluidic cards. In normal CD34+CD38− cells 36 ABC transporters were expressed, 22 of these displayed significant higher expression in the CD34+CD38− cell fraction compared to the CD34+CD38+ cell fraction. In addition to the known stem cell transporters (ABCB1, ABCC1 and ABCG2) these differential expressed genes included many members not previously associated with stem cell biology. In AML the ABC transporter expression profile was largely conserved, including expression of all 13 known drug transporters. These data suggest an important role for many ABC transporters in hematopoietic stem cell biology. In addition, the preferential expression of a high number of drug transport related transporters predicts that broad spectrum inhibition of ABC transporters is likely to be required for CD34+38− stem cell eradication in AML. This approach will, apart from affecting the leukemic stem cells, equally affect the normal stem cells.
Phenotyping of Neoplastic (CD34+/CD38−/CD123+) Stem Cells in Myeloid Malignancies Reveals Expression of Multiple Molecular Targets.
Authors: Florian Stefan , Sonneck Karoline , Hauswirth Alexander W. , Krauth Maria-Theresa , Sperr Wolfgang R. , Valent Peter 
Published: April 2020

Abstract
Recent data suggest that myeloid neoplasms are organized hierarchically in terms of self renewal and maturation of early progenitor cells, similar to normal myelopoiesis. In acute myeloid leukemia (AML), the NOD/SCID mouse-repopulating leukemic stem cells usually co-express CD123 with CD34, but lack CD38. So far, however, little is known about expression of other markers and targets on these progenitors. In the present study, expression of target antigens on CD34+/CD38− cells was analyzed by multicolor flow cytometry in patients with AML (n=18), myelodysplastic syndromes (MDS, n=6), chronic myeloid leukemia (CML, n=8), systemic mastocytosis (SM, n=9), and normal bone marrow (n=5). The IL-3Ra chain (CD123) was found to be expressed on CD34+/CD38− cells in a majority of all patients in all disease-categories. Independent of the type of disease, the vast majority of these stem cells also co-expressed aminopeptidase-N (CD13) and the target receptor CD44 in all patients. CD34+/CD38− progenitor cells expressed variable amounts of the Mylotarg® receptor CD33, KIT (CD117), HLA-DR, and AC133 (CD133). With regard to AC133, two distinct subpopulations of progenitor cells were detected in many cases, namely a CD133+ and a clearly CD133- cell-fraction. In patients with AML, the levels of CD33 varied from patient to patient with a broad range of reactivity, whereas in most patients with MDS, CML, and SM, CD33 was found to be consistently expressed on most progenitors. In most patients, neoplastic stem cells did not express substantial amounts of the GM-CSF receptor alpha chain (CD116), Thy-1 (CD90), E-NPP3 (CD203c), MDR-1 (CD243), or PAR-2. In the normal bone marrow, CD34+/CD38− cells co-expressed CD13, CD44 and CD45, but did not express CD33, CD116, or CD123. In conclusion, neoplastic stem cells in various myeloid neoplasms appear to express a similar phenotype including target receptors such as CD13, CD33, and CD44. These antigens may thus be attractive targets of therapy in AML. However, since many of these targets are not expressed on all stem cells in all patients, the elimination of the entire clone may require combinations of targeted antibodies or use of additional drugs. In other cases (CD13, CD44, CD45), the target antigen is also expressed on normal stem cells, so that targeted therapy is likely to be an ablative maneuver and thus would require a combined stem cell transplantation approach.
The Centromeric Protein, CEN(P)-F, a Marker of Cell Proliferation Is Regulated by Hypoxia in Human Mesenchymal Stem Cells and Their Bone Marrow Stromal Progeny.
Authors: Watt Suzanne M. , Hale Sarah , Baban Dilair , Roubelakis Maria , Moukayed Meis , Lim Jaeseung , Ryan Dacey J. , Davies Kay , Harris Adrian L. , Martin-Rendon Enca 
Published: April 2020

Abstract
Human bone marrow mesenchymal stem cells (MSC) are multipotent progenitors that generate osteoblasts, chondrocytes, adipocytes, myoblasts and the bone marrow stromal cells that support hematopoiesis. Although the bone marrow microenvironment is hypoxic, little is known about the maintenance and response of MSC and their bone marrow stromal progeny to hypoxia. Using cDNA microarray hybridization technologies, we show, for the first time, that a total of 231 mRNAs in cultured MSCs are regulated by short exposures (4–48hrs) to hypoxia. These include known hypoxia-responsive genes, such as BHLHB2, PGK1, GLUT-1 and VEGF. Interestingly, we demonstrate that a significant proportion of genes involved in cell growth, proliferation or survival are also regulated by hypoxia in these cells. Amongst these, we have identified the centromeric protein CENP-F as a novel gene up-regulated in cultured MSCs by hypoxia. This up-regulation results in an increased level of CENP-F protein. The hypoxic stimulus also enhances cell division in the bone marrow MSC-derived stromal cells, resulting in a doubling in cell number within 24hrs. This contrasts with the effects of hypoxia on mature endothelial cells (EC), where recruitment into cell cycle is unchanged. Our demonstration that hypoxia promotes cell division and cell proliferation in the bone marrow stromal progeny of human MSC may have wider implications for the regulated growth and survival of other stem cells in hypoxic microenvironmental niches.
Mesenchymal Stem Cells (MSCs) Suppress T Cell Proliferation by a Novel NO-stat5 Dependent Mechanism.
Authors: Sato Kazuya , Ozaki Katsutoshi , Oh Iekuni , Hatanaka Keiko , Nagai Tadashi , Muroi Kazuo , Ozawa Keiya 
Published: April 2020

Abstract
Mesenchymal stem cells (MSCs) are attractive source for regenerative therapy as they have been shown to be capable of differentiating into adipocytes, chondrocytes, osteoblasts, myocytes, cardiomyocytes, and neural precursors. MSCs have also been shown to suppress T cell proliferation in vitro and were reported to be effective as a treatment for acute graft-versus-host disease (GVHD) but the underlying molecular mechanisms for T cell suppression are uncertain. So far, TGF-β, HGF, and PGE2 were shown to be candidates as molecules causing the suppression. To address the molecular mechanisms, we used primary mouse MSCs derived from bone marrow cells and CFSE (carboxyfluorescein diacetate succinimidyl ester) or thymidine uptake for T cell proliferation assay. Co-culture of MSCs inhibited T cell proliferation induced by PMA plus Ionomycin, suggesting that TCR and signaling molecules interacting with TCR such as Lck and ZAP70 are not involved and that downstream signals of PMA plus Ionomycin are essential for the suppression by MSCs. The proliferation of either purified CD4 or CD8 cells induced by PMA plus Ionomycin was also inhibited by co-culture with MSCs, indicating MSCs suppression is active on both CD4 and CD8 cells. Stat5 phosphorylation in activated T cells was suppressed by co-culture with MSCs. Induction of cell-cycle promoting proteins such as CDK6, Cyclin D2, and Cyclin E by mitogenic stimulation were inhibited and suppression of a cell-cycle inhibitor, Kip1, was abolished. A previous report showed that T cells from stat5 deficient mice failed to induce cell-cycle promoting proteins and were not be able to proliferate on the stimulation through TCR. It was also reported that Nitric Oxide (NO) suppressed stat5 phosphorylation. Taken together with these reports, we hypothesized that NO is another candidate for the cause of suppression. In fact, NO synthase inhibitor (N-nitro-L-arginine methyl ester) recovered T cell proliferation from the suppression by MSCs in a dose-dependent manner. The amount of NO production and the strength of T cell suppression were parallel and dependent on the number of MSCs. MSCs blocked production of IFNγ but induction of T cell activation markers such as CD25 and CD69 and production of IL-2 were unaffected as reported. Our data suggest that MSCs block stat5 phosphorylation by production of NO, resulting in that T cells can neither proliferate nor produce high level of IFNγ. Here we demonstrate a new critical NO-stat5 dependent mechanism for T cell suppression by MSCs.
Six Courses of Intensified CHOP Plus G-CSF Compared to Eight Courses of Standard CHOP in Patients with Intermediate-Risk Aggressive Non-Hodgkin Lymphoma Results of a Prospective Randomized HOVON Trial.
Authors: Verdonck Leo F. , van Imhoff Gustaaf W. , Raemakers John M.M. , Verhoef Gregor , Kramer Mark H.H. , Ossenkoppele Gert J. , Doorduijn Jeanette K. , Meulendijks Ine A.M.H. , van Wieringen Wessel N. , Sonneveld Pieter 
Published: April 2020

Abstract
Between November 1994 and February 2004 a total of 513 untreated patients with aggressive intermediate-risk NHL, aged 16–65 years, were recruited to the randomized HOVON-26 trial comparing intensified (I)CHOP plus G-CSF (given every 2 weeks for 6 courses) with standard CHOP (given every 3 weeks for 8 courses). I-CHOP included doxorubicin (D) at 70 mg/m2 and cyclophosphamide (C) at 1000 mg/m2 together with standard dose vincristine and prednisone. So, dose intensity (mg/m2/week) for both D and C was doubled in the I-CHOP arm compared to CHOP, whereas the cumulative dose of D and C was similar in both treatment arms. Intermediate-risk aggressive NHL was defined (HOVON criteria) as stage II disease with increased LDH and stage III/IV disease with normal LDH, corresponding to the low-intermediate (L-i) and high-intermediate (H-i) risk groups of the age-adjusted (aa) IPI score. The endpoints were CR rate, overall (OS), disease-free (DFS) and event-free survival (EFS).
Of 513 patients, 477 were eligible of whom 239 were randomized to CHOP and 238 to I-CHOP. Patients were well balanced for all risk factors. According to the aa-IPI score, in the L-i group 161 patients received CHOP and 137 I-CHOP, whereas in the H-i group 68 patients received CHOP and 80 patients I-CHOP.
As of July 1st 2005, median follow-up of all patients still alive is 50 months. Ninety-nine patients in the CHOP arm died and 85 patients in the I-CHOP arm died. NHL was the main cause of death in both treatment arms (76 and 61 patients, respectively). Treatment-related mortality (8%) was similar in both arms, however WHO grade 4 side-effects occurred twice as often (20 vs 10 events) in the I-CHOP arm. I-CHOP did result in better CR rate and 5 years estimates of OS, DFS, and EFS, but the differences were not significant (table 1). However, although not originally planned, subgroup analysis showed improved OS, DFS, and EFS in the low-intermediate group (table 2).
Although I-CHOP was not significantly better than CHOP in intermediate-risk aggressive NHL patients, subgroup analysis indicates a possible improvement in outcome for patients with low-intermediate risk aggressive NHL. Another advantage of I-CHOP might be that therapy duration is markedly reduced.
Table 1. CR rate and 5 yrs OS, DFS and EFS of intermediate-risk patients (HOVON criteria) CR rate OS DFS EFS CHOP 49% 57% 52% 25% I-CHOP 53% 62% 63% 32% p-value (log-rank) 0.41 (chi-square) 0.19 0.19 0.17 Table 2. 5 yrs OS, DFS and EFS in low- intermediate and high-intermediate risk groups (aa-IPI score) Low-intermediate aa IPI High-intermediate aa IPI OS DFS EFS OS DFS EFS CHOP 60% 48% 24% 52% 54% 25% I-CHOP 68% 66% 36% 52% 53% 28% p-value (log-rank) 0.06 0.03 0.03 0.78 0.85 0.84
Transplant Outcome in Acute Myeloid but Not Acute Lymphocytic Leukemia Following T Cell Depleted Allogeneic Stem Cell Transplantation May Be Favorably Influenced by Early Recovery of NK Cells.
Authors: Savani Bipin N. , Montero Aldemar , Mielke Stephan , Rezvani Katayoun , Shenoy Aarthi , Childs Richard , Barrett A. John 
Published: April 2020

Abstract
Between 05/1994 and 07/2005, 78 patients with acute leukemia (AML=49; ALL=29) received a T cell depleted stem cell transplant from an HLA-identical sibling, with 1–2 infusions of donor T cells on days 30–100. The conditioning regimen included 12–13.5 Gy total body irradiation (TBI) and cyclophosphamide (Cy) with or without fludarabine. Cyclosporine (CSA) was given as the sole GVHD prophylaxis after transplant. The stem cell source was peripheral blood in 75 and bone marrow in 3. The T cell dose ranged between 0.2 to 2 × 105CD3+ cells/kg. AML and ALL patients were distributed equally among transplant regimen and T cell dose variations. The median CD34 cell dose was 5.4 × 106 cells/kg, (5.4 for AML, and 5.5 for ALL; p=0.50). Actuarial disease free survival (DFS), transplant related mortality (TRM) and relapse rate was 42±8, 20±6 and 40±9 % for AML and 22±10; 35±19 and 64±11% for ALL patients. The median day 30 post-transplantation lymphocyte count (LC30) for AML patients was 418/μl (31–2105) and 778/μl (265–3295) for ALL patients; p=0.063). In multivariate analysis, LC30 emerged as an independent risk factor for transplant outcome in AML but not ALL patients (see figure). In AML patients LC30 above median was associated with superior DFS, lower TRM and lower relapse. CD34 dose and LC30 were highly correlated for AML and ALL analyzed together or separately. Lymphocyte subset analysis of 20 post-transplant samples on day 30 identified a large proportion of CD56+16+ natural killer (NK) cells (median 44%, range 15–86%). Since early post-transplant NK cells are immature and unregulated by KIR-expression they may exert anti-host reactivity. The more favorable outcome for AML but not ALL patients achieving higher lymphocyte counts early post-transplant recapitulates the myeloid leukemia-restricted benefit from superior NK cell recovery observed in AML but not ALL recipients of mismatched T cell depleted stem cell transplants reported by Ruggeri et al (Blood.1999;94:333–9).
Figure Figure
Definition of Cytochrome P450 1A2 as a Target Autoantigen in the Hepatitic Variant of Hepatic Chronic Graft-Versus-Host Disease.
Authors: Mullighan Charles G. , Bogdanos Dimitrios , Vergani Diego , Bardy Peter G. 
Published: April 2020

Abstract
Chronic graft-versus-host disease (cGVHD) of the liver mimicking autoimmune hepatitis is an increasingly well recognized complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). The target antigens and pathogenesis of this complication are poorly understood. We describe a case of hepatitic cGVHD in which cytochrome P450 1A2 (CYP1A2) was identified as a target antigen. A 47 year old male received an HLA matched sibling transplant for acute myeloid leukemia with normal cytogenetics following 3 cycles of idarubicin, cytarabine and etoposide remission induction therapy. Conditioning was with fludarabine 50mg/m2 and cyclophosphamide 120 mg/m2. Both donor and recipient were CMV IgG positive. Standard cyclosporin and short course methotrexate GVHD prophylaxis was used. Due to mixed lymphoid chimerism, cyclosporin was tapered from D94, and a donor leukocyte infusion (DLI) given at D124. One month following DLI, concomitant with attainment of 100% donor lymphoid chimerism, the patient developed florid oral changes of cGVHD and striking elevation of hepatic transaminases (ALT and AST peaked at 3604 (R 0–55 IU/L) amd 2241 (R 0–45 IU/L) respectively) with relatively minor elevation in hepatic ductal markers (GGT 327, R 0–60 IU/L). Liver biopsy was consistent with autoimmune hepatitis. Multiple recipient serum samples at this time demonstrated the presence of a high titre anti-liver microsomal (LM) antibody (titre 1:1280 by indirect immunofluorescence) reminiscent of hydralazine-induced hepatitis and immunofixed an antigen of approximately 51 kDa by immunoblotting using human liver homogenate. Neither pre-transplant recipient nor donor sera were reactive. Immunoblotting for a panel of 13 CYP450 antigens demonstrated that the antibody was specific for CYP1A2. This specificity was confirmed with blocking and inhibition experiments. Sequencing of the entire coding region of the CYP1A2 gene did not identify any coding variants present in the recipient but not the donor. Oral prednisolone resulted in brisk improvement in hepatitis and oral cGVHD, and was ceased at 12 months post transplant. The patient remains well with no further GVHD over 2 years post transplant. This is the first description of a target antigen in hepatitic cGVHD. The mechanism is unclear, but does not appear to be a CYP1A2 polymorphism acting as a minor histocompatibility antigen. Induction of hepatic autommune-like responses have been described in liver transplant recipients receiving calcineurin inhibitors, and it is possible that cyclosporin facilitated an autoimmune-type response in this case. Characterisation of autoantibody responses in additional cases of hepatitic cGVHD will be of great interest.
Figure Figure
Measuring Multiple Symptoms and Inflammatory Cytokines Related to Acute GVHD in AML/MDS Patients under Allogeneic BMT.
Authors: Giralt Sergio , Cleeland Charles S. , Reuben James , Lee Baing-Ning , Williams Lori , Mobley Gary , Wang Xin S. 
Published: April 2020

Abstract
The objective of this study was to prospectively assess the relationship between symptom severity and changes in inflammatory cytokines around the time of diagnosis of acute GVHD in AML/MDS patients during the first 100 days of allo-BMT. Methods: Weekly symptom assessment with the M. D. Anderson Symptom Inventory was done for 30 patients over time of BMT. A panel of inflammatory cytokines (IL-1ra, IL-6, IL-8, IL-10, IL-12, TNF-a) was assayed at multiple time points (pre BMT, conditioning, BMT day, Day 1, nadir, Day 7, 14, 21, 28, 60, 90–100, also including days 1, 3, 5, 10 and 15 after diagnosis of a GVHD for the 26 patients who developed it). Results: Of the 26 patients, 69% had grade 1 GVHD, 27% had grade 2, and 4% had grade 3. 42% were treated with 2mg/kg of steroids, 31% with topical steriods. Figure 1 displays Loess curves that show the multiple symptom pattern for the 26 GVHD patients. These nonspecific symptoms reached their highest levels before the day of diagnosis. Immediate symptom reduction after diagnosis was most likely caused by administration of steroids. The Lowess curves in Figure 2 show the changes in serum cytokines levels pre- and post-GVHD (log based 10). IL-8 was significantly correlated with some symptoms before and after GVHD (Table 1). Conclusion: During development of GVHD in the acute phase of allo-BMT, there is an observed relationship of certain symptom severity and increased or decreased inflammatory cytokine levels in this pilot study. Further study is warranted for a causal link between general sickness and specific cytokines during human GVHD development.
Table 1. Correlations between symptoms severity and cytokines Levels before and after the Development of GVHD in the First 100 Days after Allo-BMT IL-8 Pre-GVHD Shortness of breath r = −0.708, P&lt;.01 Dry mouth r = 0.667, P&lt;.05 Sadness r = 0.631, P&lt;.05 Post-GVHD Sleep Disturbance r = 0.564, P&lt;.05 Drowsiness r = −0.565, P&lt;.05 Vomiting r = −0.558, P&lt;.05 Figure Figure Figure Figure
Gene Signatures Predictive of Outcome in Higher Risk Childhood Acute Lymphoblastic Leukemia (ALL).
Authors: Moskowitz Naomi P. , Bhojwani Deepa , Kang Huining , Min Dong-Joon , Potter Jeffrey , Harvey Richard , Seibel Nita L. , Raetz Elizabeth , Sather Harland N. , Hunger Stephen P. , Willman Cheryl L. , Carroll William L. 
Published: April 2020

Abstract
The use of risk adapted therapy has improved outcome for children with ALL and assessment of early response (ER) has been used to identify a subset of patients who benefit from augmented treatment. In spite of these advances, the majority of treatment failures occur in a “good risk” subset indicating the critical need for better predictors of outcome. We performed oligonucleotide microarrays on 99 B-precursor patients treated on CCG 1961 for high risk ALL to identify: 1) genetic pathways that play a role in early response to therapy (42 day 7 rapid (R)ER, 40 slow (S)ER) and 2) a signature that would predict event free survival (28 pts &gt; 4 yr CCR, 31 relapses &lt; 3 yrs). RNA was extracted and amplified before hybridization to Affymetrix U133 Plus 2.0 microarrays representing over 47,000 transcripts. Data were normalized, filtered and then analyzed by both unsupervised and supervised methods. We identified a robust signature that predicted SER at day 7 (79% of cases in a test set) but not RER thus limiting its utility in risk stratification. Furthermore, we also identified a highly significant set of genes that correlated with SER (M2/M3) at day 14. Importantly, expression of 47 probe sets were significantly different in patients who were in CCR for more than 4 years compared to cases that experienced a relapse within the first 3 years (FDR ≤0.05%). Logistic regression indicated that each of the 47 probe sets has significant prognostic value beyond that contained in the clinical covariates (including gender, age and WBC count). A major subset of these genes were involved in signal transduction (MAP3K3, YWHAZ, PCTK2, TGFBR1, PTPRE, IFNGR2, HSPA8). In a separate validation analysis, subsets of genes were selected using logistic regression with backward, forward and stepwise variable selection and the models proved to be significantly predictive of response in the independent expression data set of 220 patients generated previously using a smaller probe set with U95Av2 microarrays (Mosquera-Caro et al, Blood 2003, 102(11)). Interestingly, the genetic models superseded clinical co-variates such as age, WBC and gender. Finally, to assess further the predictive accuracy of these gene signatures, we randomly divided the data set into a training set (2/3 cases) that was used to develop a signature predictive of relapse that was tested for accuracy in the remaining cases (1/3 cases). This was repeated three times. On average, 77% of patients in CCR and 73% of patients who relapsed were accurately classified in the test set. With further validation these signatures of outcome (not early response) will allow for more precise identification of patients who are at very high risk for relapse and may provide a mechanistic understanding of why patients fail therapy.