bluuebunny/crossref_metadata_2025_split huggingface dataset:
Common Submicroscopic Genomic Imbalances Accompany the Ph Chromosome at Diagnosis in Chronic Myeloid Leukemia
Authors: Khorashad Jamshid S , De Melo Valeria A , Fiegler Heike , Gerrard Gareth , Brudzevsky Dan , Marin David , Apperley Jane F. , Goldman John M , Foroni Letizia , Reid Alistair G 
Published: April 2020

Abstract
Despite the presence of a consistent genetic abnormality, CML patients display considerable clinical heterogeneity, the basis of which is poorly understood. We therefore used a novel ultra-high-resolution genomic screening assay to search for additional acquired genomic abnormalities that might explain this clinical heterogeneity and help to assess prognosis for individual patients. Comparative genomic hybridisation (CGH) was performed with a 2.1 million oligonucleotide array on DNA extracted from presentation samples of 20 previously untreated CML patients. Each DNA sample from diagnosis was competitively hybridized against the same patient’s remission sample, which avoided detection of constitutional polymorphic copy number variants (CNVs). For representative copy number aberrations (CNAs) the CGH result was confirmed by fluorescence in situ hybridization or quantitative real-time PCR. All 20 CML patient samples harbored detectable genomic imbalances with a median of 16 CNAs per patient (range: 4–165). Of the 846 CNAs detected 679 (80%) were amplifications and 167 (20%) were deletions. 462 CNAs (55%) involved at least one known gene. The median size of CNAs was 58kb (range 9kb – 12Mb). One hundred and twenty seven different genomic regions were aberrant in 2 or more patients in the cohort. Of these recurrent CNAs, amplifications of the FOXD4L4, MAPK8IP1, DUSP1, PSTPIP1 and PBEF1 genes were among the most frequently detected; they were present in 3, 4, 5, 6 and 15 patients respectively. FOXD4L4 is a myeloid trancription factor previously implicated in leukemogenesis, and PSTPIP1 has been shown to be involved in dephosphorylation of ABL. MAPK81P1 and DUSP1 are both involved in the MAPK pathway, while PBEF1 is known to play a role in the NAD anti-apoptotic pathway. The presence of multiple genomic lesions at diagnosis supports the notion of an increased level of genomic instability in CML cells and raises the possibility that one or more aberrations in addition to BCR-ABL may dictate the CML phenotype. The considerable range in the number of CNAs identified at diagnosis may reflect a differing level of genomic instability in different patients. Furthermore the presence of same genetic imbalance in more than one patient suggests a role for specific genes in addition to BCR-ABL in the pathogenesis or progression of CML.
Circulating B-cell chronic lymphocytic leukemia cells display impaired migration to lymph nodes due to reduced LFA-1 expression
Authors: Hartmann Tanja Nicole , Grabovsky Valentin , Wang Wei , Desch Petra , Wollner Stefan , Binsky Inbal , Vallon-Eberhard Alexandra , Sapoznikow Anita , Haran Michal , Shachar Idit , Burger Meike , Honczarenko Marek , Greil Richard , Alon Ronen 
Published: April 2020

Abstract
Homing to secondary lymphoid organs and re-entry to bone marrow (BM) are central aspects of leukemic pathophysiology. We investigated the roles of the two major lymphocyte integrins LFA-1 and VLA-4 on B-cell chronic lymphocytic leukaemia (CLL) cells in these processes. We found that CLL cells expressed significantly reduced LFA-1 due to low beta2 integrin transcripts and displayed diminished adhesiveness to ICAM-1-expressing endothelium in vitro. VLA-4 expression was heterogenous but underwent rapid activation by the BM chemokine CXCL12. Nevertheless, CLL cells failed to transmigrate across VCAM-1, ICAM-1 and CXCL12 expressing endothelium due to their deficient LFA-1 expression. Furthermore, when injected into tail veins of immunodeficient mice, normal B cells rapidly homed to lymph nodes (LNs) in a LFA-1 dependent manner whereas CLL cells did not. CLL alike normal B lymphocytes used VLA-4 rather than LFA-1 to reenter the BM. In contrast, both normal and CLL B cells homed to mice spleen in an LFA-1- and VLA-4-independent manner. Our results suggest that CLL cells are deficient in LFA-1-dependent trafficking to LNs but residual subsets can still re-enter the BM. Integrin blocking could be therefore an efficient strategy to prevent circulating CLL cells from reaching prosurvival niches in LNs and BM and directing these cells to the spleen.
Direct Fluorescent Visualization of HSC Competitive Repopulation in Transparent Adult Zebrafish
Authors: Li Pulin , Zon Leonard I 
Published: April 2020

Abstract
Transplanted hematopoietic stem cells (HSCs) home to the marrow space, engraft and self-renew. The process of engraftment is difficult to study in most vertebrates because it is impossible to visualize the engraftment directly. Here we have developed competitive transplantation of fluorescent donor marrow samples into a completely transparent adult zebrafish called casper. This allows the direct visualization of engraftment and the process of HSC competition. Transplantation of zebrafish whole kidney marrow (WKM) cells, containing HSCs, progenitors and mature blood cells, can rescue lethally irradiated recipient fish and repopulate all the blood lineages. We used different GFP-/DsRed-labeled transgenic fish as donors in WKM transplantation, such as the myeloid specific Tg(mpo:GFP), or the ubiquitous expression of Tg(β-actin:GFP), and Red GloFish® to study the kinetics of engraftment. Engraftment can be visualized with a simple fluorescent microscope, and was evident in the recipient kidney as early as 2 weeks post transplant (wpt). By 4 wpt, mature hematopoietic cells differentiated from donor HSCs or progenitors can be observed in the recipient circulation as well as in epidermis as tissue residential macrophages. At 8 wpt, donor-derived lymphoid cells also repopulate the recipient immune system. The fluorescence intensity of engrafted cells was quantified by image analysis software and positively correlated with the engraftment efficiency quantified by FACS. Using Red GloFish® as competitor donor and Tg(β-actin:GFP) as test donor, we were able to see the competition between the two donors both by fluorescence intensity analysis and FACS analysis. To test the sensitivity of this competitive transplant system to the alteration of the ratios between the two colors, we increased the ratio of the test vs. competitor donor from 1:3 to 1:2 to 1:1. At 4wpt, in the 1:3 ratio group, only 14% of the recipients had stronger GFP intensity than the red (n=7), while the rest had weaker or equivalent GFP intensity than the red; however, this percentage increased to 43% and 50% in the 1:2 (n=7) and 1:1(n=8) groups respectively. Therefore, the relative engraftment ratio can be fast detected in vivo by direct fluorescent visualization with the transparent zebrafish recipients. Next, we treated the test donor cells with 16,16-methyl prostaglandin E2 (dmPGE2) or DMSO for 2 hours in vitro at room temperature before transplant. The engraftment efficiency of the dmPGE2 group dramatically increased by at least 2 folds compared with the DMSO group. The colors of competitor and test donors were switched, and the increase of engraftment efficiency by dmPGE2 treatment of the test donor was still evident. In conclusion, direct visualization of competitive marrow repopulation greatly facilitates the developmental, physiological or pathological study of HSCs and progenitors and should provide a new platform for the assay of specific genes or chemicals that could alter homing, engraftment or self-renewal.
A Closer Look at Cellular Iron Metabolism in IRP2 Deficient Erythroblasts.
Authors: Schranzhofer Matthias , Schifrer Manfred , Galy Bruno , Hentze Matthias W. , Mullner Ernst W. , Ponka Prem 
Published: April 2020

Abstract
Developing red blood cells are the major consumers of body iron which is indispensable for the enormous production of heme for hemoglobin synthesis. The uptake of iron occurs via binding of iron-loaded transferrin to its cognate receptor (TfR). Thereafter the iron is shuttled to the mitochondria where it is incorporated into protoporphyrin IX to form heme. Excess iron is enclosed within the iron storage protein ferritin. Coordinated control between iron uptake and storage is mainly achieved by the post-transcriptional regulation of TfR1 and ferritin synthesis by the iron regulatory proteins IRP1 and IRP2. Recently, two groups independently created mice lacking either IRP1 or IRP2 and showed that only IRP2 deficient mice developed microcytic hypochromic anemia. Both groups observed a reduction in TfR1 protein expression levels in the developing red blood cells of IRP2 knockout animals and suggested that the decrease in receptor levels is responsible for the development of anemia.
For a more detailed analysis of how the loss of IRP2 expression influences iron metabolism and hemoglobinization during terminal erythroid differentiation, we isolated CFU-E-like erythroid cells from mouse fetal liver of wild type, IRP1 and IRP2 knock out animals. In vitro cultivation of these primary erythroid cells and their synchronous induction for differentiation allowed us to study their cellular iron metabolism at different time points. We analyzed the extent of hemoglobinization and cell size as well as the expression of ferritin and TfR1 during various stages of erythroid differentiation in IRP1, IRP2 and wild type cells. In agreement with the published phenotype of microcytic hypochromic anemia, only erythroblasts lacking IRP2 exhibited a reduction in hemoglobinization and showed a significant increase in ferritin protein levels before and after induction of differentiation. In contrast, TfR1 protein expression levels on the cell surface were significantly decreased in IRP2 deficient cells until 24h of differentiation, but converged with those of wild type cells at 48h of differentiation at the time point at which hemoglobinization is fully in progress. Moreover, measurement of 59Fe uptake and its cellular distribution showed that there is significantly more 59Fe located in cytosolic ferritin of IRP2 knock out cells at all time points compared to their wild type counterpart. In summary, these results suggest that not only the reduced expression of TfR1, but also the up-regulation of ferritin, play important roles in the development of anemic phenotype in IRP2 knock out mice.
This work was supported by the Canadian Institutes of Health Research and the Canadian Blood Services.
T Regulatory Cells in Graft-Versus-Host Disease of the Skin
Authors: Azar Jose , Billings Steven , Schwartz Jennifer E. , Liu Yunlong , Yu Menggang , Nelson Robert P. 
Published: April 2020

Abstract
FoxP3+ CD25+ T regulatory (Treg) cells are known to be present in normal skin. Treg cell functional deficiency resulting in loss of suppression of activation, differentiation or expansion of effector T cells could conceivably contribute to the pathophysiology of graft-versus- host disease (GVHD). Rezvani et al. suggest that levels of peripheral blood Treg cells in donors and recipients may predict the risk of acute GVHD. Rieger et al. found a significantly lower number of Treg cells in human colonic biopsies with GVHD versus CMV colitis or normal samples. There are no reports that evaluate tissue Treg cells in human skin affected by GVHD. This study was conducted to evaluate the distribution of Treg cells in skin affected by acute GVHD versus chronic GVHD. Archived samples from patients previously reported by routine histopathological methods as acute or chronic GVHD were collected, coded and entered into a database. For immunostaining, 4-μm thick serial sections were cut and deparaffinized. Immunohistochemical stains for CD4 (Neomarkers, 1:20), CD8 (Dako, prediluted), CD25 (Dako, 1:100), and Foxp3 (Serotec, 1:250) were performed using standard techniques. The dermatopathologist evaluating the samples was blinded to the clinical outcomes. Results were scored as 0 (&lt;10% of lymphocytes positive), 1+ (10–25% positive), 2+ (26–50% positive), and 3+ (≥50% positive). Fourteen patients with acute skin GVHD and seventeen with chronic GVHD were identified from a database of patients who had undergone nonmyeloablative allogeneic peripheral blood transplantation in the recent past. The average scores for each immunostain were calculated and are summarized in table 1. The average FoxP3 score in acute GVHD specimens was significantly lower than that in chronic GVHD specimens (average, 0.57 versus 1.41; p-value = 0.011). The average scores of CD3, CD4, CD8 and CD25 immunostains were not significantly different between acute and chronic GVHD biopsies. These findings represent the initial observation of a distinction between the distribution of regulatory T cells in acute and chronic GVHD of the skin. These observations should be confirmed in a larger sample, supported by functional assays of Treg cells, and correlated with clinical outcomes. Such studies may help to elucidate the role of Treg cells in acute and chronic skin GVHD.
Table 1.
Average score CD 3 CD 4 CD 8 CD 25 Fox P3 Acute GVHD 2.93 2.21 2.14 0.29 0.57 Chronic GVHD 2.82 2.44 2.24 0.71 1.41 p-value 1.00 0.42 0.43 0.34 0.011
Immunological Evaluation after Haematopoietic Stem Cell Transplantation for B–Thalassemia Major
Authors: Marziali Marco , Isgro Antonella , Gaziev Javid , Fraboni Daniela , Polchi Paola , Sodani Pietro , Alfieri Cecilia , Paciaroni Katia , Gallucci Cristiano , Lucarelli Guido 
Published: April 2020

Abstract
Background: Haematopoietic stem cell transplantation (HSCT) remains the only curative option for patients with thalassemia. After HSCT is particularly important to recover a complete immune reconstitution for prevention of opportunistic infections.
Methods: The immunological phenotype of peripheral blood mononuclear cells in 110 consecutively subjects after HLA identical HSCT performed for b thalassemia were studied. CD3+CD4+, CD3+CD8+, CD3−CD19+, CD3−CD16+CD56+ were evaluated by Flow Cytometry at 6 and 12 months after HSCT in 70 patients followed. In a part of patients CD4 naïve T cells was also evaluated at 12 months after HSCT.
Results: at 6 months after HSCT we observed reduction of the mean of CD4 T cell percentage in patients respect normal value: in mean 17, 5 % ± 9 vs 45 % ± 10 respectively. After 1 year the rate of CD4+ T-cell population progressively increase: 24 ± 9 at 12 months vs 17, 5 % ± 9 at 6 months, but the mean of CD4+ T-cell count was persistently lower than in controls. The mean of CD8 T cell is stable during the follow up and the CD4/C8 ratio decreased. The mean of the CD19 increase during the follow up (4, 5 % at 6 months vs 17% at 12 months). At 1 year after HSCT the percentage of NK (CD3−CD16+CD56+) was normalized. At 1 year after HSCT reduction of naïve CD4 T cell we observed.
Discussion. At 1 year after HSCT the immunological reconstitution is not still complete. The time and the extent of immune reconstitution depend on several factors. It is very important to continue the immune surveillance after HSTC and to evaluate other immunological parameters (i.e. thymic output) for to prevent infectious complications and elaborate an appropriate therapeutic strategy.
Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) in Stem Cell Transplant Recipients: Response to Immune Enhancement in Early Versus Late Serious Post-Transplant Infections
Authors: Safdar Amar , Georgescu Georgina , Rodriguez Gilhen , Champlin Richard 
Published: April 2020

Abstract
Background: The impact of GM-CSF therapy in SCT recipients with serious infection is not clear. We evaluated feasibility of GM-CSF therapy in transplant patients.
Methods: Forty-seven patients with severe infections who had received 7 or more doses of GM-CSF between 2000 and 2006 were evaluated. Response was considered at the end of GM-CSF therapy in patients with complete or partial response (CR/PR). All values are given as median ± s.d. Serious infections were defined as systemic/invasive infections requiring hospitalization.
Results: Age was 39 ± 18 years; 83% received allogeneic SCT, and 70% had leukemia. GVHD was noted in 36%; 60% of SCT recipients had cancer recurrence and 11% had graft failure. The APACHE II score was 15 ± 4. GM-CSF was given for 13 ± 12 (range, 7–57) doses. In only 1 of 5 patients with GM-CSF related adverse events (AEs), growth factor was discontinued due to persistent bone pain. Patients who received GM-CSF during late (&gt;180 days) infections had higher CR/PR versus cytokine therapy for patients with early post-transplant infections (63% vs 35%, respectively; P = 0.06). Factors associated with treatment failure included: prior high-dose steroids (43% vs 4% in CP/PR; P = 0.01) multiple concurrent infections (65% vs 33% in CR/PR; P = 0.03), critical care unit stay (44% vs 5% in CR/PR; P = 0.001), mechanical ventilation (35% vs 4% in CR/PR; P = 0.01), serum bilirubin &gt; 2mg/dL (30% vs 0%) in CR/PR; P = 0.004) and neutropenia (50% vs 17% in CR/PR: P = 0.02). Neutropenia was 25 ± 19 days in responders vs 34 ± 42 days in patients who failed treatment (P = 0.08). By logistic regression analysis, presence of multiple concurrent infections was associated with higher probability of death within 12 weeks of infection diagnosis (odds ratio, 4.76; 95% confidence interval, 1.1–20.5; P = 0.03).
Conclusions: Treatment with GM-CSF was tolerated without serious AEs and may improve outcomes in patients with late post transplant infections.
Prognostic Factors Associated with Outcomes of RIC and MAC Hematopoietic Stem Cell Transplantation: An Indication for the Selection of Conditioning
Authors: Kiyokawa Tetsuyuki , Nagakura Shoichi , Hidaka Michihiro , Yano Takahiro , Sunami Kazutaka , Okamura Seiichi , Uike Naokuni , Hamaguchi Motohiro , Matsui Takahiro , Horibe Keizo , Sato Kensaku , Kawakita Toshiro , Kawano Fumio 
Published: April 2020

Abstract
BACKGROUND: Despite recent increase of reduced intensity conditioning (RIC) transplantation, mortality rates after RIC and myeloabrative conditioning (MAC) HSCT remain high and cannot accurately predicted.
OBJECTIVE: To determine the value of pre-transplant factors in predicting one year overall survival of allogeneic HSCT with RIC and MAC. Optimizing conditioning selection based on pre-transplant prognostic factors may further improve results of allogeneic HSCT.
PATIENTS AND METHODS: A retrospective review of 415 consecutive allogeneic HSCT was performed with attention to mortality and pre-transplant factors in five hematopoietic cell transplantation centers between 2000 and 2005. Patients underwent transplantation from sibling (n=275) or unrelated donors (n=140) with MAC (n=247) or RIC (n=168). Main outcomes and prognostic factors were analyzed in multivariable analyses (a logistic regression model) with RIC and MAC.
RESULTS: Mortality before one year occurred in 252 of 415(60.1%) transplant recipients; 137 of 247(55.4%) with MAC and 115 of 168(68.5%) with RIC. Multivariate analysis identified specific and distinct prognostic factors with one year overall survival with MAC (CR/non-CR, performance status, albumin level, creatinine leve and a contain of Ara-C with preconditioning regimens) and with RIC (PS, albumin level, a contain of fuldarabin)(Table). These factors were useful for estimating survival rate of recipient after one year of transplantation (survival estimating equation).
CONCLUSION: Our results may provide indication for a better choice of conditioning regimen according to pre-transplant prognostic factors
Figure Figure Figure Figure
Prolonged Survival in Adults with Acute Lymphoblastic Leukemia (ALL) Following Reduced Intensity Conditioning and Allogeneic Transplantation
Authors: Bachanova Veronika , Weisdorf Daniel J. , Brunstein Claudio , Wagner John , Verneris Michael 
Published: April 2020

Abstract
Reduced intensity conditioning (RIC) and allogeneic hematopoietic cell transplantation (HCT) for adult patients with ALL may improve the tolerance and outcomes following allografting. We studied 22 consecutive patients (2001–2008) who received RIC HCT. Indications for RIC regimens included: age &gt; 55 years (for matched related donor [MRD], &gt;45 years for unrelated or umbilical cord [UCB]), Karnofsky index &lt;80%, impaired organ function or prior autologous/allogeneic HCT. The median age was 49 years (24–68), median diagnostic WBC 20×109/L (1.4–248) and median comorbidity index of 3 (range: 0–8) at HCT. All patients had high risk disease, including 14 patients (64%) with Ph+ ALL, 10 patients (46%) in 2nd or subsequent remission. Five patients (23%) were treated with tyrosine kinase inhibitors pre-HCT. Uniform conditioning (Cyclophosphamide 50 mg/kg; Flu 200 mg/m2; total body irradiation (TBI) 200cGy, n=21) and graft versus host disease (GvHD) prophylaxis (Cyclosporin/MMF) was followed by MRD (n=4) or UCB donor (n=18) grafts. All patients had sustained donor neutrophil engraftment (median 10 days, range 0–28) and reached 100% donor chimerism at median day 23 post-transplantation (range 14–99 days). At 3 years, treatment related morality relapse and overall survival (OS) were 27% (95% confidence interval [CI]: 9–45%), 36% (95% CI: 14–58%) and 50% (95% CI: 27–73%), respectively. There were no relapses beyond 2 years. After a median follow-up of 33 months (range 5–77) the cumulative incidence of acute II–IV, III–IV and chronic graft versus host disease were 55%, 20% and 45%. Lower TRM for HCT in CR1 vs ≥ CR2 (8% vs. 50%; p&lt;0.04) led to improved 3 year OS in the CR1 cohort (81% vs. 15% (p&lt;0.01)). HCT using either MRD or UCB resulted in similar risks of GVHD, relapse and OS. These data suggest that RIC allogeneic HCT using either related or UCB donors for adults with ALL can result in modest TRM, and only limited risks of relapse and promising leukemia-free survival. Further study, especially for patients in CR1, will better define the utility of this approach for patients unsuited for more intensive, myeloablative conditioning.