bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation of Ldb1 Stability and Transcriptional Complex Assembly by Single-Stranded DNA-Binding Protein SSDP2.
Authors: Xu Zhixiong , Meng Xianzhang , Cai Ying , Nagarajan Lalitha , Brandt Stephen J. 
Published: April 2020

Abstract
The LIM domain-binding protein Ldb1 is known to form higher order complexes with LIM-homeodomain and LIM-only (LMO) proteins to regulate diverse developmental programs, including hematopoiesis. The level of Ldb1 is critical for its cellular roles, and its turnover is regulated by the E3 ubiquitin ligase RLIM. Single-stranded DNA-binding protein (SSDP), an Ldb1-interacting partner, is an essential gene for embryonic development and has been shown to regulate axis formation in Xenopus and wing development in Drosophila through Ldb1; however, the mechanisms by which SSDPs regulate these and other developmental programs are still obscure. We previously reported that a DNA-binding complex containing the basic helix-loop-helix protein TAL1/SCL, its DNA-binding partner E47, zinc finger protein GATA-1, LIM domain protein LMO2, and Ldb1 stimulates Protein 4.2 (P4.2) transcription in erythroid progenitors through tandem E box-GATA elements in the gene’s proximal promoter. We have now established that SSDP2 is associated with this complex (by supershift analysis) and occupies the promoter of this gene (by chromatin immunoprecipitation analysis) in murine erythroleukemia (MEL) cells. Further, enforced expression of SSDP2 in these cells stimulated P4.2 reporter activity and accumulation of P4.2 and beta-globin mRNAs, and cotransfection of SSDP2 with the five originally identified components of this complex further increased promoter activity in reporter analysis. Importantly, overexpression of SSDP2 in MEL cells significantly increased Ldb1 protein half-life and steady-state levels of Ldb1 and LMO2 protein. This effect on Ldb1 stability required the Ldb1-interacting domain of SSDP2, consisting of its first 94 amino acids (SSDP2(1–94)), and was also observed in Cos7L and CHO cells. We showed, in addition, that SSDP2 or SSDP2(1–94), but not an Ldb1 interaction-defective mutant, prevented RLIM-mediated degradation of both Ldb1 and LMO2 in transfected cells, that SSDP2 protection of LMO2 degradation required Ldb1, and that SSDP2 directly inhibited RLIM-mediated ubiquitination of Ldb1. Immunoprecipitation analysis revealed that overexpression of SSDP2 or SSDP2(1–94) significantly decreased interaction between Ldb1 and RLIM. Finally, SSDP2 protein expression in differentiating MEL cells paralleled this multi-protein DNA-binding activity and overexpression of SSDP2 in these cells dramatically increased E box-GATA DNA-binding activity, with maximal formation of the ternary complex requiring coexpression of SSDP2, Ldb1, and LMO2. Together, these studies reveal a positive role for SSDP2 in erythroid gene expression and identify a biochemical function for SSDP2 in regulating Ldb1 stability and transcriptional complex assembly. The mechanism mediating Ldb1 stabilization appears to involve competitive inhibition of RLIM interaction with Ldb1.
Thrombotic Thrombocytopenic Purpura after Allogeneic Hematopoietic Stem Cell Transplantation: A Complication Still Frequent and Responsible of Severe Outcome.
Authors: Uderzo Cornelio , Sangalli Haidi , Bonanomi Sonia , Morreale Giuseppe , Lanino Edoardo , Annaloro Claudio , Volpe Aldo Della , Alessandrino Paolo , Lazzarino Mario , Renoldi Mila , Ferrari Paolo , Iacobelli Massimo , Busca Alessandro , Locatelli Franco , Balduzzi Adriana , Rovelli Attilio 
Published: April 2020

Abstract
Introduction
Thrombotic thrombocytopenic purpura (TTP) might costitute a severe complication after hematopoietic stem cell transplantation (HSCT) with a variable incidence depending on different diagnostic criteria adopted in the past.
Patients and Methods
The current study enrolled 551 consecutive patients (pts) (314 males, median age 29 years) undergoing HSCT from January 2000 to April 2005 from HLA match (m) or mismatch (mm) family donor (350/551) and unrelated donor (201/551) for malignant (501/551) or non malignant diseases (50/551). The diagnosis of TTP was performed on the basis of homogeneous five clinical and seven laboratory criteria. Univariate (by chi square test and Fisher’s exact test) or multivariate (by logistic regression method) analyses were performed to evaluate the effect of some candidate risk factors on both TTP occurrence and outcome.
Results
Sixty four of 551 pts presented TTP (11,6%) at a median time of 47 days post-HSCT; 59/64 were affected by malignant and 5/64 by non malignant diseases. Multivariate analysis showed GVHD &gt;II° (p=0,0001), stem cell donors (m/mm unrelated or mm related) (p=0,024), female gender (p=0,0186),TBI-based conditioning regimen (p=0,03) and status of hematological remission (p=0,03) as factors influencing the occurrence of TTP. Only three predicting factors for TTP outcome have been demonstrated statistically significative by multivariate analysis: age (p=0,0186), stem cell donors (m/mm unrelated or mm related) (p=0,01) and TTP index (p=0,018). The TTP mortality rate was 40% while the outcome after TTP diagnosis was partially influenced by TTP treatment. In particular, Defibrotide seemed to be a promising drug in univariate analysis compared with alternative therapies for 34 alive and good responders pts out of 64 TTP pts.
Conclusions
The study underlines the possibility to identify those patients who are more prone to develop post-HSCT TTP, complication still frequent (11,6%) when homogeneous diagnostic criteria are adopted. TTP outcome seems to be conditioned by some peculiar risk factors as adult age, m/mm unrelated or mm related donors HSCTs. Prospective randomized therapeutic trials, focusing on the possible advantage of Defibrotide versus other treatments, should be encouraged.
Efficacy of Soluble TNFα Inhibitor (Etanercept) for Treatment of Steroid Refractory Acute Graft Versus Host Disease (GVHD) Post Allogeneic Stem Cell Transplantation (SCT).
Authors: Kennedy Glen A. , Butler Jason , Morton James , Hill Geoff R. , Western Robyn , Durrant Simon 
Published: April 2020

Abstract
Aims: To review the efficacy of the soluble TNFα inhibitor etanercept in treatment of steroid refractory acute GVHD.
Methods: All patients who received etanercept for treatment of biopsy proven steroid refractory acute GVHD at our institution were retrospectively reviewed. GVHD was graded as per Seattle criteria. Standard GVHD prophylaxis post SCT was cyclosporine plus either methotrexate (D1–11) or mycophenolate mofetil (MMF). Standard 1st line therapy used for grade 2–4 GVHD was methylprednisolone 2mg/kg/day; standard 2nd line therapy for steroid refractory GVHD was anti-thymocyte globulin (ATG) + tacrolimus +/− MMF. Etanercept was used either after no-response (NR) &gt;1mth after administration of both 1st line and 2nd line therapies (Group 1), or included as part of standard 2nd line therapy, starting within approximately 2wks of commencing ATG (Group 2). Etanercept dose was 0.4mg/kg (max 25mg) SC twice per week for 4wks, then once per week for 4wks. GVHD response was assessed at end of etanercept therapy. Standard antibiotic prophylaxis included valaciclovir (for HSV / VZV), bactrim / pentamidine (for PCP) and fluconazole (for fungal infection).
Results: In total 17 patients had received etanercept therapy, including 3 with grade 2, 7 with grade 3 and 7 with grade 4 GVHD. Predominant organ affected by GVHD was gut in 9 cases, skin in 5 and combination gut and skin in 3. Of the 3 patients treated in group 1, no responses and no survivors occurred. Group 2 included 14 patients, in whom responses were 10 CR (72%), 1 PR (7%) and 3 NR (21%), with 7 patients (50%) alive at median FU of 6mths (range 3–30mths) post commencing etanercept. For the entire cohort, OS at 12mths is 35%. Causes of death included progressive GVHD in 4 patients and infective complications in 6. Overall infective complications observed included bacterial sepsis (6/17), CMV reactivation (8/17, including CMV pneumonitis in 1), other viral infections (3/17), new, possible or proven aspergillosis (3/17) and progression of previously documented fungal infection (2/17).
Conclusions: Our experience suggests that when used in combination with ATG, etanercept has significant activity in the treatment of steroid refractory acute GVHD, with high response rates and significant survival observed. Better strategies to reduce infectious complications in this setting are needed.
Chronic Graft-Versus-Host Disease after Allogenaic Bone Marrow Transplantation in Japan: Identification of Risk Factors and Prognostication.
Authors: Atsuta Yoshiko , Suzuki Ritsuro , Yamamoto Kazuhito , Terakura Seitaro , Iida Hiroatsu , Kohno Akio , Naoe Tomoki , Sao Hiroshi , Kodera Yoshihisa , Morishima Yasuo , Hamajima Nobuyuki , Morishita Yoshihisa 
Published: April 2020

Abstract
Objective: Chronic graft-versus-host disease (cGVHD) is an important complication of allogeneic hematopoietic stem cell transplantation (HSCT) involving various organs such as skin, eyes, mouth, liver, lung, and gastrointestinal tract, which is associated with major disability and diminished quality of life. There have been several studies to explore risk and prognostic factors in Western countries. However, because the genetic homogeneity is different among the ethnics, such models are not assured to be applicable for Japanese patients. To identify the incidence and risk factors of cGVHD, and prognostic factors for patients who developed cGVHD, we analyzed a series of patients.
Methods: The incidence and prognostic factors for cGVHD were evaluated for 255 patients who survived more than 100 days after bone marrow transplantation (BMT) in Nagoya BMT Group.
Results: One hundred and nineteen (47%) developed cGVHD. Prior acute GVHD (grade 2–4) and use of unrelated donor were significantly associated with the onset of cGVHD. Systemic immunosuppressive therapies were administered to 86 patients (78%), 42 (38%) of whom required secondary therapies. Presence of cGVHD did not have an impact on mortality (hazard ratio [HR]=0.89; 95% confidence interval [CI], 0.59–1.3). Proposed prognostic models from Western countries, Akpek’s score and Lee’s grading system did not successfully classify our patients. Three factors at diagnosis were associated with cGVHD-specific survival; i.e. presence of infection (HR=4.1; 95%CI, 1.6–10.3), ongoing use of corticosteroid at the onset of cGVHD (HR=3.9; 95%CI, 1.7–9.1), and a Karnofsky performance score&lt;80 (HR=4.7; 95%CI, 2.0–11.3). The probability of cGVHD-specific survival at 4 years was 79% (95%CI, 70–86%).
Conclusion: The severity and death rate of Japanese patients with cGVHD was lower than those for populations in Western countries, which might be the result of greater genetic homogeneity of Japanese ethnics. For prognostication, a different model was needed to identify high-risk patients.
Calcitonin A Genotype Is Associated with Risk of Acute Graft-Versus-Host Disease Following Allogeneic Bone Marrow Transplantation for Children with Acute Myeloid Leukemia in First Remission.
Authors: Goyal Rakesh K. , Fairfull Liane , Gerbing Robert B. , Alonzo4 Todd A. , Meshinchi4 Soheil , Lange4 Beverly J. , Woods4 William J. , Ferrell Robert E. 
Published: April 2020

Abstract
Background: Procalcitonin (PCT) is a 13 kD 116-amino acid prohormone. Elevated serum levels of procalcitonin have been noted in patients with bacterial infections such as septicemia, pneumonia and immune-mediated conditions like graft-versus-host disease (GvHD) and organ rejection. In this study, we sought to examine if a single base pair polymorphism (a T to C transition) at position −624 of the calcitonin A gene (CALCA) was associated with risk of acute GvHD.
Subjects: 113 children (median age 10 y; range 0.38 – 19.0 y) with de novo acute myeloid leukemia (AML) treated on recent Children’s Oncology Group protocols 2891, 2941 or 2961. All of these patients received matched sibling bone marrow transplant (BMT) in first clinical remission. Ablative conditioning consisted of busulfan and cyclophosphamide. All patients received methotrexate until day 100 for GvHD prophylaxis.
Methods: All clinical DNA specimens were subjected to whole genome amplification. Genotyping was performed using the 5′-nuclease (TaqMan) assay on the ABI 7700 DNA analyzer (Applied Biosystems).
Results: Observed CALCA -624 T&gt;C genotypes in 113 patients were: TT = 52 (46%), TC = 43 (38%) and CC = 18 (16%); T allele = 73 (65%) and C allele = 40 (35%). There were no significant differences in patient characteristics (age, gender, race, study 2891/2941/2961, white blood count at diagnosis, FAB type, or cytogenetics) between the TT vs. combined TC or CC groups. Overall, acute GvHD grade I–IV (Seattle criteria) was observed in 55/113 patients (49%). Nineteen out of 52 (37%) with TT, while 21 out of 43 (49%) with TC, and 15 out of 18 (83%) with CC CALCA genotypes developed acute GvHD. Alternatively, acute GvHD was observed in 37% patients with common TT genotype and in 59% patients with variant TC or CC genotype (p=0.023). The odds ratio for developing acute GvHD (grade I – IV) for children with TC or CC genotypes was 2.5 (CI 1.17, 5.33) compared with those with TT genotype. The difference in observed acute GvHD remained significant when the analyses were limited to more uniform subgroups: 54 patients on single trial, 2961 only (p=0.028) and 42 whites only on 2961 trial (p=0.028). Severe acute GvHD (grade III – IV) was seen in 6/52 (12%) patients with TT vs. 6/61 (10%) in those with TC or CC genotypes; this difference was not statistically significant (p =0.771). There were no significant differences in day +100 transplant related deaths, engraftment, relapse rate or overall survival. Plans: Association between CALCA genotypes and infectious complications will be also analyzed when clinical data abstraction is completed. CALCA −1752 C&gt;G and 3740 A&gt;G genotyping is in progress and clinical correlative analyses will be repeated at genotype and haplotype levels.
Outcome with the Hyper-CVAD and Imatinib Mesylate Regimen in Philadelphia (Ph) Positive Acute Lymphocytic Leukemia (ALL).
Authors: Thomas Deborah A. , Faderl Stefan , Cortes Jorge , O’Brien Susan , Giles Francis , Garcia-Manero Guillermo , Kornblau Steven , Andreeff Michael , Beran Miloslav , Verstovsek Srdan , Pierce Sherry , Letvak Laurie , Salvado August , Champlin Richard , Kantarjian Hagop 
Published: April 2020

Abstract
In Ph+ ALL, complete remission (CR) rates with intensive chemotherapy such hyper-CVAD is 90%, but most patients (pts) relapse within a median time of 16 months [Kantarjian et al, JCO18:547, 2000; Kantarjian et al, Cancer101:2788, 2004]. Single agent therapy with the tyrosine kinase inhibitor imatinib mesylate in relapsed or refractory Ph+ ALL or chronic myelogenous leukemia in lymphoid blast phase yielded CR rates of 20% with rapid disease recurrence. A phase II clinical trial of concurrent hyper-CVAD and imatinib was designed to improve these results, with the initial regimen of imatinib 400 mg orally daily days 1–14 of each course (fractionated cyclophosphamide, vincristine [VCR], doxorubicin and dexamethasone alternating with high dose methotrexate and cytarabine) followed by imatinib, VCR and prednisone maintenance with intensifications months 6 and 13. Allogeneic stem cell transplant (SCT) was performed in CR if feasible. Preliminary results of first 20 pts treated were encouraging [Thomas et al, Blood103:4396, 2004]. Recent modifications included increasing dosing of imatinib to 600 mg daily days 1–14 of course 1, then daily if tolerated with courses 2–8. Maintenance was extended to 24 months with imatinib indefinitely. To date, 43 pts with Ph+ ALL have been treated from April 2004 to July 2005. Thirty-six pts had active disease, either untreated (n=31) or refractory (n=1) to one induction course without imatinib; 7 pts were in CR after one induction course without imatinib. Of 35 evaluable pts, 33 (94%) achieved CR (1 induction death, 1 failed to meet platelet criteria for CR). Median days to response was 21 days. 13 pts underwent allogeneic SCT within a median of 3 months from start of therapy (range, 1–12). After a median follow-up of 3 yrs (range 1–48 months), 1 primary refractory pt relapsed at 12 months, 1 de novo pt had isolated CNS relapse, 2 pts relapsed after allogeneic SCT (no post SCT imatinib) and 2 pts changed therapy for persistent Ph+ metaphases (1 relapsing). Deaths in CR included 5 older pts without allogeneic SCT (1 osteomyelitis, 1 mucormycosis, 1 C. difficile colitis, 1 sudden death, 1 GNR sepsis) and 4 pts after allogeneic SCT (3 graft-versus-host disease, 1 GNR sepsis). Outcome with the hyper-CVAD and imatinib regimen continues to demonstrate favorable disease-free survival rates compared with hyper-CVAD alone, particularly for the de novo group. Use of higher dose imatinib concurrently appears to be feasible. Molecular response rates appear to be improved with the higher dose imatinib; additional accrual will be required to assess impact of modifications, including role of allogeneic SCT.
Myeloid Lineage-Restricted Progenitors Contribute to Vascular Endothelium.
Authors: Bailey Alexis S. , Willenbring Holger , Jiang Shuguang , Schroeder David A. , Anderson Daniel A. , Grompe Markus , Wong Melissa H. , Fleming William H. 
Published: April 2020

Abstract
Endothelial progenitors of hematopoietic origin have a role in promoting recovery from vascular injury. Recently we have shown that following radiation damage, transplanted hematopoietic stem cells (HSCs) give rise to endothelial cell outcomes at the clonal level. This raises the question of whether prospectively identifiable HSC progeny have endothelial cell activity. Using β-galactosidase and EGFP expression as donor markers, we show that lineage-restricted myeloid progenitors can give rise to endothelial cells. Transplanted common myeloid progenitors (CMPs) and granulocyte/macrophage progenitors (GMPs) gave rise to similar numbers of endothelial cells in liver portal vein branches. Donor-derived endothelial cells expressing functional endothelial markers including CD31, von Willebrand factor (vWF) and Tie-2 were clearly distinguished from mature hematopoietic cells by the absence of CD45 and F4/80 expression. Confocal microscopy showed the co-localization of donor and endothelial cell marker expression in individual cells so that endothelial cells could be discriminated from perivascular cells. To evaluate cell fusion as a potential mechanism for the emergence of endothelial cells of hematopoietic origin, HSCs transgenic for EGFP and Tie-2-Cre recombinase were transplanted into ROSA26 reporter (R26R) mice. Activation of β-galactosidase reporter gene expression, indicative of cell fusion with a donor-derived hematopoietic cell, could be detected in hepatocytes but not in endothelial cells. However, due to the tissue-specificity of the Tie-2 promoter, endothelial fusion products in which the donor nucleus had failed to activate the Tie-2-driven Cre recombinase would have also lacked detectable reporter gene expression. To ensure that the absence of β-galactosidase expression in the donor-derived endothelial cells was not due to insufficient nuclear reprogramming, HSCs transgenic for EGFP and R26R were transplanted into Tie-2-Cre recipients. Donor-derived endothelial cells were again negative for both β-galactosidase expression and activity. To determine whether radiation injury is required for the incorporation of donor cells into the endothelium, a parabiosis model was utilized. Both long-term donor-derived endothelial cell outcomes and multilineage hematopoiesis were detected in parabiotic mice. These results demonstrate that CMPs and further lineage-restricted GMPs are capable of differentiating into endothelial cells in the absence of cell fusion. Importantly, radiation injury is not required for endothelial cell engraftment, indicating that circulating progenitors contribute to angiogenesis during steady-state conditions.
CUL-4A Short Hairpin RNA (shRNA) Impairs Normal Hematopoiesis of Human Cord Blood CD34+ Cells In Vitro and In Vivo.
Authors: Shieh Jae Hung , Zhang Jianxuan , Lu Xiaoling , Zhou Pengbo , Moore Malcolm A.S. 
Published: April 2020

Abstract
The CUL-4A ubiquitination machinery regulates the stability of HOXA9 by promoting its ubiquinitation and proteasome-dependent degradation. Our previous study showed the perturbation of CUL-4A biosysthesis in 32D cells by RNA-mediated interference increased an accumulation of HOXA9, and impaired G-CSF-induced terminal differentiation to granulocytes. HOXA9 is preferentially expressed in primitive hematopoietic cells and downregulates with differentiation. In this study, a lentiviral vector expressed both CUL-4A shRNA and GFP was constructed and was used to transduce cord blood (CB) CD34+ cells (GFP-shCUL-4A-CD34+ cells). The ability of GFP-shCUL-4A-CD34+ cells to form CFC colonies, to differentiate into myeloid, erythroid, B-lymphoid and T-lymphoid cells in vitro, and to engraft in irradiated NOD/SCID mice was evaluated. Our results showed that knockdown of CUL-4A m-RNA dramatically inhibited hematopoisis both in vivo and in vitro. In the CFC assay, FACS-sorted GFPbright-shCUL-4A-CD34+ cell formed fewer colonies (5%) that were smaller size (&lt;500 cells/colonies) relative to progeny of GFPbright-CD34+ cells. In contrast, FACS-sorted GFPdim-shCUL-4A-CD34+ cells produced only 50% less colonies than the control GFPdim-CD34+ cells. However, GFPdim-shCUL-4A-CD34+ cells had 30% more colonies with &lt;500 cells/colony but 82% less colonies with &gt;500 cells/colony comparing to that of GFPdim-CD34+ cells. After transplanting the GFP-shCUL-4A-CD34+ cells into irradiated NOD/SCID mice for 6 weeks, GFP-shCUL-4A-CD45+ cells decreased more than 80% in the mouse bone marrow. These human GFP-shCUL-4A CD45 + cells were capable of forming GFP+-CFC but with &gt;80% reduction in their potency. No reduction in % of GFP-CD45+ cells and GFP+-CFC was observed in the GFP-CD34+ cells group pre- and post-transplantation. The in vitro culture systems showed that knockdown of CUL-4A mRNA in CB CD34+ cells resulted in the developmental impairment in monocytic, megakaryocytic, erythroid, B lymphoid and T-lymphoid lineages. This impairment is a time- and shCUL-4A dose-dependent process. Further study using cell cycle analysis indicated that transduction of CUL-4A shRNA to HL-60 or CD34+ cells induced apoptosis. Our results, for the first time, demonstrate an essential role of the CUL-4A ubiquitin ligase in maintaining cell survival to ensure proper differentiation of normal human stem and progenitor cells into different hematopoietic lineages both in vivo and in vitro.