bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fyn Specifically Regulates the Activity of Red Cell Glucose-6-Phosphate-Dehydrogenase
Authors: Mattè Alessandro , Lupo Francesca , Tibaldi Elena , Di Paolo Maria Luisa , Andrea Carpentieri , Pucci Pietro , Brunati Anna Maria , Cesaro Luca , Turrini Francesco , Manzo Saúl Gómez , Choi Soo Young , Quino Jaime Marcial , Kim Dae Won , Pantaleo Antonella , An Xiuli , Federti Enrica , Iatcenko Iana , Cappellini Maria Domenica , Forni Gian Luca , De Franceschi Lucia 
Published: November 2019

Fyn is a tyrosine kinase belonging to the Src family (Src-Family-Kinase, SFK), ubiquitously expressed. Previously, we reported that Fyn is important in stress erythropoiesis. Here, we show that in red cells Fyn specifically stimulates G6PD activity, resulting in a 3-fold increase of the enzyme catalytic activity (kcat) by phosphorylating tyrosine (Tyr)-401. We found Tyr-401 on G6PD as functional target of Fyn in normal human red blood cells (RBC), being undetectable in G6PD deficient RBCs (G6PD-Mediterranean and G6PD-Genova). Indeed, Tyr-401 is located to a region of the G6PD molecule critical for the formation of the enzymatically active dimer. Amino acid replacements in this region are mostly associated with a chronic hemolysis phenotype. Using mutagenesis approach, we demonstrated that the phosphorylation status of Tyr401 modulates the interaction of G6PD with G6P and stabilizes G6PD in a catalytically more efficient conformation. RBCs from Fyn-/-mice are defective in G6PD activity, resulting in increased susceptibility to primaquine-induced intravascular hemolysis. This negatively affected the recycling of reduced Prx2 in response to oxidative stress, indicating that defective G6PD phosphorylation impairs defense against oxidation. In human RBCs, we confirm the involvement of the thioredoxin/Prx2 system in the increase vulnerability of G6PD deficient RBCs to oxidation. In conclusion, our data suggest that Fyn is an oxidative radical sensor, and that Fyn-mediated Tyr-401 phosphorylation, by increasing G6PD activity, plays an important role in the physiology of RBCs. Failure of G6PD activation by this mechanism may be a major limiting factor in the ability of G6PD deficient RBCs to withstand oxidative stress.

Disclosures
Cappellini: Genzyme/Sanofi: Honoraria, Membership on an entity's Board of Directors or advisory committees; Vifor Pharmaceutical: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees; Celgene Corporation: Honoraria; CRISPR Therapeutics: Membership on an entity's Board of Directors or advisory committees. Forni:Novartis, Iron chelation: Research Funding; Celgene, Erithropoiesis Stimulation: Research Funding; Roche, Erithropoiesis Stimulation: Research Funding; BlueBirdBio: Consultancy.
Eltrombopag Treatment for Patients with Thrombocytopenia Post-Transplantation Showed a Good Chance of Platelet Recovery
Authors: Gao Fei , Zhao Yanmin , Huang He 
Published: November 2019

Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) dramatically improves the outcomes of patients with hematologic disorders. However, thrombocytopenia post-transplantation is a common and severe complication, which not only diminishes the quality of life but also leads to fatal bleeding events. Blood transfusion could not be a permanent solution and it seldom achieves satisfactory results. Eltrombopag (EPAG), a novel oral thrombopoietin receptor agonist (TPO-RA), has been shown promising effects in thrombocytopenia caused by immune thrombocytopenic purpura (ITP) and severe aplastic anemia (SAA). Yet, the effectiveness of EPAG for patients with thrombocytopenia post-transplantation still needs to be discovered. From September 2017 to April 2019, twenty-two patients were diagnosed as thrombocytopenia post-transplantation in our center, including 10 patients with poor graft function (PGF) and 12 patients with secondary failure of platelet recovery (SFPR). Since insensitive to traditional treatment such as thrombopoietin and intravenous immunoglobin, all these patients received the EPAG treatment. So far, 16 (72.7%) patients achieved hematopoietic overall recovery (OR) and 11(50.0%) patients were complete recovery (CR). 12(75.0%) of the responded patients discontinued or tapered the drug and 9 patients (56.3%) successfully maintained their best response. Among 16 responded patients, the median time to achieve PR status was 33 (range, 7-105) days and the median time of CR was 67 (range, 40-175) days. Interestingly, 11 (91.7%) patients from SFPR subgroup achieved OR, while only 5 (50%) patients from PGF subgroup achieved OR. Besides, patients with normal spleen size (n=16) experience a higher CR rate than splenomegaly patients (n=8) (10 vs 1, 71.4% vs 12.5%). On univariate and multivariate analysis, PGF was identified as the independent risk factor of OR (P=0.050) and splenomegaly was identified as the independent risk factor of CR (P=0.019). Interestingly, 3 patients with enlarged spleen received ruxolitinib to minimize the spleen size during the EPAG treatment. Accompanied by the reduction in spleen size, the platelet counts increased gradually and all these 3 patients achieved OR. In contrast, only 2 of the other 5 splenomegaly patients who without ruxolitinib achieved OR. In conclusion, EPAG treatment for patients with thrombocytopenia post-transplantation showed a good chance of platelet recovery.

Disclosures
No relevant conflicts of interest to declare.
Allogeneic Hematopoietic Stem Cell Transplantation for Therapy-Related Myelodysplastic Syndromes and Acute Myeloid Leukemia
Authors: Metheny Leland , Saber Wael , Weisdorf Daniel J. , de Lima Marcos , Callander Natalie Scott , Litzow Mark , Kebriaei Partow , Hall Aric C. , Sandmaier Brenda M , Zhang Mei-Jie , Wang Hai-Lin , Bo-Subait Khalid 
Published: November 2019

Patients who develop therapy-related myelodysplastic syndromes (t-MDSs) or acute myeloid leukemia (t-AML) following prior chemotherapy or an autologous hematopoietic cell transplant (HCT), have a poor prognosis. An earlier analysis of allogeneic HCT (n=868, 1990-2004) showed 5-year overall survival (OS) and disease free survival (DFS) were 22% and 21% (reference). Furthermore, older age, graft source other than a HLA-matched sibling or HLA well matched unrelated donor (URD), t-AML not in remission or advanced t-MDS, and poor risk cytogenetics were associated with worse outcomes.
Modern supportive care and conditioning regimens including reduced intensity (RIC) regimens may have improved outcomes. Therefore we analyzed 1531 HCT for t-AML (n = 772) or t-MDS (n = 759) performed from 2000 to 2014.
The median age at transplant was 52 (18-77) for t-AML and 59 (18-74) for t-MDS. Eleven percent of patient with t-AML and 24% of patient with t-MDS had a prior autologous transplant. A myeloablative regimen was used for conditioning in 61% of patients with t-AML and 49% of patients with t-MDS (Table 1).
For t-AML, the 5-year non-relapse mortality (NRM) and relapse rates (RR) were 33% (95% confidence interval [CI], 30-36) and 43% (40-47), respectively. For t-MDS, the 5 year TRM and RR were 32% (29-36) and 46% (43-50), respectively. Five year DFS and OS were 24% (21-27) and 25% (22-28), respectively, for patients with t-AML. Five year DFS and OS were 21% (18-24) and 27% (23-31), respectively, for patients with t-MDS.
In multivariate analysis, t-AML OS and DFS was worse for: previous autologous transplant, relapsed disease at HCT, intermediate or adverse cytogenetics, partially matched and cord blood grafts, and RIC. For t-MDS: age greater than 60, previous autologous transplant, very high IPSS score, and use of HLA-partially matched unrelated grafts.
A significant minority of patients with t-AML and t-MDS can achieve long-term remission with allogeneic HCT, yet risks of both NRM and relapse are still excessive. Relapse is the major cause of treatment failure.

Disclosures
Metheny: Takeda: Speakers Bureau; Incyte: Speakers Bureau. Weisdorf:Incyte: Research Funding; Fate Therapeutics: Consultancy; Pharmacyclics: Consultancy. Kebriaei:Jazz: Consultancy; Amgen: Research Funding; Pfizer: Honoraria; Kite: Honoraria.
Association of Nucleophosmin-1 (NPM1) Expression with Leucocyte Count, Blast Count, and Gingiva Hypertrophy in Acute Myeloid Leukemia De Novo Patients in General Hospital of Dr M Djamil Padang Indonesia
Authors: Wahid Irza , Atmakusuma Tubagus Djumhana 
Published: November 2019

Acute myeloid leukemia (AML) is a malignant disease derived from hematopoietic system characterized by neoplastic transformation of myeloid progenitor cell, present as excessive proliferation and differentiation alteration leading to inhibition of myeloid maturation. From various studies, incident and prevalence of AML these days is increasing in morbidity and mortality. This make AML known with poor prognosis. Nucleophosmin-1 (NPM-1) was established by WHO revision 2016 as prognostic gene marker in AML. Nucleophosmin-1 gene is mutated in 35-60% of AML cases. Recently, molecular biologic development suggests that prognostic gene examination is needed to reduce morbidity and mortality of AML patients. If myeloid progenitor cell encounters carcinogenic/leukemogenic stresses, mutation of several genes could happen. NPM1 gene is a likely mutated gene and predominantly located in nucleolus. Overexpression of NPM1 will promote activities of tumor suppressors like P53, ARF and MDM2. Interactions of those tumor suppressors promote proliferation, differentiation, cell survival, apoptosis and DNA repair, which give a better prognosis in AML patients.
To study about association of NPM1 expression with leucocyte count, blast count, and gingiva hypertrophy in AML de novo patients in general hospital of Dr M Djamil Padang, Indonesia.
Analytic observational study with cross-sectional approach to 25 adult AML patients from Policlinic and Inpatient Installation of Dr M Djamil Hospital from March to August 2018 who met inclusion and exclusion criteria. Expression of NPM1 was measured with qRT-PCR.
We found significant increase of NPM1 expression in AML in the amount of 22.62x107 copies/ml. There was a significant very strong correlation between NPM1 expression and leucocyte count (p<0.05 & r=0.870) and blast count (p<0.05 & r=0.828). There was also significant differences of NPM1 expression in AML patients with gingival hypertrophy and without gingival hypertrophy (31.3 x 107 copies/ml dan 9.6 x 107 copies/ml).
There was a significant very strong correlation between NPM1 expression with leucocyte count and blast count. There was significant increases of NPM1 expression in AML patients with gingival hypertrophy.
Keywords: acute myeloid leukemia, Nucleophosmin-1, leucocyte, blast, gingival hypertrophy

Disclosures
No relevant conflicts of interest to declare.
ABL Kinase Domain Mutation in Chronic Myeloid Leukemia - Compliance to Imatinib Matters
Authors: PK Jayachandran , Ganesan Trivadi S , Mehra Nikita , Kannan Krishnarathinam , Dhanushkodi Manikandan , Radhakrishnan Venkatraman , Rajan Arun Kumar , Girishchandra Joshi Archit , Selvarajan Gangothri , Karunakaran Parathan , Sagar Tenali Gnana 
Published: November 2019

Background: Imatinib resistance mutation analysis (IRMA) or abl kinase domain mutation analysis is performed in patients with Chronic myeloid leukemia (CML) whenever the response to treatment is inadequate. We have analyzed the reports of IRMA at our centre.
Methods: The clinical details of 71 patients with CML on Imatinib, who underwent IRMA testing during the period of January 2017 to March 2019 were collected from the patient records and analyzed. IRMA was performed for failure or warning or progression, anytime during the course of treatment. IRMA was done by either Sanger sequencing (n=45) or next generation sequencing (n=26, Illumina, NGS). The associations between variables were tested using Chi - Square test.
Results: Median age at diagnosis of 71 patients was 44 years (Range 18 - 71 years). Males constituted 70% (n=50). At diagnosis, 92% (n=65) of patients were in chronic phase and the remainder were in accelerated phase (n=4) or blast crisis (n=2). Mutations in the abl kinase domain were detected in 26 patients (37%). Next Generation Sequencing (NGS) could identify more mutations (13/26 - 50%) compared to conventional Sanger Sequencing (13/45 - 29%), but the difference was not significant (p=0.07). NGS could identify three or more mutations in 5 patients in contrast to Sanger. All the mutations detected were those previously described except for an insertion of 35bp near the C-Terminal which was identified in 3 patients. E459K translocation was identified in 6 patients. E355G translocation was identified in 4 patients. F359V, M351T, Y253H, G250E, H396R, T315I translocations were identified in 3 patients each. Patients who were not compliant to therapy had increased frequency of mutations (14/26 - 54%) compared to those who were compliant (12/45 - 27%), which was significantly different (p=0.02). Patients who had loss of complete hematological response (CHR) had significantly higher frequency of mutations (14/21- 67%) compared to other reasons for performing the test (p=0.001). Patients who had failure to achieve targets at various time points had a significantly lower frequency of mutations (4/23 - 17%, p=0.02) compared to other reasons for performing the test.
Conclusion: Patients who were not compliant for treatment were more likely to have mutations. Loss of CHR showed an increased frequency compared to other reasons. NGS could identify mutations in more number of patients. NGS identified numerically higher mutations in patients. Larger prospective data are needed to confirm these observations.
Table

Disclosures
No relevant conflicts of interest to declare.
Implementation of the Peripherally Inserted Central Venous Catheter for Patients with Hematological Malignancies in a Peruvian Cancer Center
Authors: Roque Katia , Enriquez Daniel J , Elizabeth Elera , Ortega Emperatriz , Haro Juan C , Poma Nathaly , Ponce Ricardo , Tirado-Hurtado Indira , Neciosup Silvia , Gomez Henry 
Published: November 2019

Background: A central venous access is essential during active treatment for most patients with hematological malignancies, and historically, the central inserted venous catheter (CVC) has been the preferred option in clinical practice. In the last years, the peripheral inserted central catheter (PICC) has increasingly been used as an alternative to CVC with less incidence of catheter related infections (CRI). Our aim was to compare the incidence of documented catheter related infections and mortality for patients with PICC versus CVC.
Methods: We prospectively enrolled patients who were candidates for PICC implantation from March 2018 to February 2019, and matched them with a CVC candidate patient. PICC patients were followed at our PICC line program and CVC patients with their primary oncologist team as per institutional protocol. All patients received intensive treatment at a public tertiary cancer center in Lima-Peru (Instituto Nacional de Enfermedades Neoplasicas).
Results: We included 94 cases during the study period (44 PICC and 50 CVC cases), median age was 39 years and M:F ratio was 2:1. In the PICC group, main diagnoses were acute leukemias (46%), Lymphomas (22%) and Multiple Myeloma (16%), on the other hand, in the CVC group, acute leukemias (72%), lymphoma (16%) and multiple myeloma (8%). Mean length of catheterization was 40 and 24 days for PICC and CVC, respectively. The incidence of documented CRI was lower in PICC group (12.6 versus 3.1 per 1000 days for CVC and PICC, respectively), showing a risk reduction of 75% of bacteremia (RR: 0.25, 95%CI 0.1-0.6, p<0.01). Mortality rate was 20% and 14% in CVC and PICC groups, respectively (p=0.4).
Conclusion: PICC represents a safe and effective alternative to CVC for patients with hematological malignancies in a limited source setting. An alarming high incidence of catheter related infections was noted in this study.

Disclosures
No relevant conflicts of interest to declare.
Expansion of Th/Tc17 T Cells and Loss of CX3CR1+ Macrophages Play a Critical Role in the Pathogenesis of Steroid-Refractory Acute Intestinal Gvhd
Authors: Song Qingxiao , Wang Xiaoning , Martin Paul J , Chen Yuanzhong , Zeng Defu 
Published: November 2019

Acute gut graft-versus-host disease (GVHD) is the major contributor to mortality and morbidity of allogeneic hematopoietic stem cell transplantation recipients. Steroids are the first-line therapy for acute GVHD, but approximately 30% of patients develop steroid refractory GVHD (SR-GVHD). The pathophysiology of SR-GVHD is poorly understood.
In the current studies, we explored the mechanisms of SR-GVHD, using a murine model of MHC-mismatched HCT with C57BL/6 donors and BALB/c recipients. Acute GVHD can be mediated by either Th/Tc1 or Th/Tc17 T cells. We evaluated the impact of dexamethasone (DEX) treatment on acute GVHD induced by spleen and BM cells from WT, IFN-γ-/- or RORγt-/- C57BL/6 donors. WT donor T cells give rise predominantly to Th/Tc1 cells early after HCT, but Th/Tc17 cells can expand as time goes on. IFN-γ-/- donor T cells give rise to Th/Tc17 cells but not to IFN-γ-producing Th/Tc1 cells, while RORγt-/- donor T cells give rise to Th/Tc1 cells but not to Th/Tc17 cells. BALB/c recipients were treated with single dose dexamethasone (DEX) at 5 mg/Kg on day 3 after HCT. DEX treatment effectively prevented acute GVHD in recipients given WT or RORγt-/- grafts for up to 20 days after HCT. GVHD then recurred in recipients grafted with WT cells but not in recipients grafted with RORγt-/- cells. In contrast, DEX treatment had no effect on acute GVHD in recipients grafted with IFN-γ-/- cells. Recurrence of GVHD in DEX-treated recipients grafted with WT cells and the lack of DEX effect in recipients grafted with IFN-γ-/- cells was associated with expansion of Th/Tc17 cells that produced both IL-17 and IL-22. In recipients grafted with IFN-γ-/- cells, we also observed marked reduction of donor-type CX3CR1+ intestinal macrophages that play a critical role in controlling bacterial translocation from the intestinal lumen to the liver and blood. In addition, the DEX-treated recipients showed augmentation of Th/Tc17 differentiation and exacerbation of acute GVHD, When CX3CR1GFP/GFP mice that are deficient in generating CX3CR1+ intestinal macrophages were used as donors.
These results indicate that DEX-treatment can cause expansion of Th/Tc17 and loss of protective donor-type CX3CR1+ intestinal macrophages, leading to SR-GVHD.

Disclosures
Martin: Abgenomics: Research Funding; Enlivex Therapeutics: Consultancy; Genentech: Consultancy, Other: One-time advisory board; Neovii: Other: One-time advisory board; Pfizer: Other: Data and Safety Monitoring Committee; Pharmacyclics LLC: Other: One-time advisory board; Procter and Gamble: Equity Ownership; Xenikos: Research Funding.
Tissue-Resident PSGL1loCD4+ T Cells Interact with B Cells Via PD-1 and PD-L2 in Gvhd Target Tissues to Augment Autoimmune-like Chronic Gvhd
Authors: Kong Xiaohui , Zeng Deye , Wu Xiwei , Yang Shijie , Wang Bixin , Song Qingxiao , Zhu Yongping , Salas Martha , Qin Hanjun , Nasri Ubaydah , Nakamura Ryotaro , Martin Paul J , Huang Aimin , Zeng Defu 
Published: November 2019

Chronic graft versus host disease (cGVHD) is a systemic autoimmune-like syndrome, and donor-type CD4+ T interaction with B cells plays an important role in its pathogenesis. Our recent publication has demonstrated that cGVHD pathogenesis does not require germinal center formation and is associated with expansion of extrafollicular CD44hiCD62LloPSGL1loCD4+ (PSGL1loCD4+) T cells in the spleen and GVHD target tissues. The causal role of PSGL1loCD4+ T cells in cGVHD pathogenesis remains unclear. With murine and humanized murine models, we show that both murine and human PSGL1loCD4+ T cells reside in tissues and do not circulate in the blood, and their high expression of PD-1 and ICOS and production of IL-21 suggest that they provide B cell help. In murine models, PSGL1loCD4+ T cells augment plasma cell expansion and autoantibody production via their PD-1 interaction with PD-L2 on B. In humanized mouse models, human PSGL1loCD4+ T cells from GVHD target tissues augment memory B cell differentiation into plasma cells and antibody production via their production of IL-21.These results indicate that tissue-resident PSGL1loCD4+ T cells augment autoantibody production and cGVHD pathogenesis. Targeting tissue-resident T cells may represent a novel approach for treating cGVHD.

Disclosures
Nakamura: Alexion: Other: support to a lecture at a Japan Society of Transfusion/Cellular Therapy meeting ; Kirin Kyowa: Other: support for an academic seminar in a university in Japan; Celgene: Other: support for an academic seminar in a university in Japan; Merck: Membership on an entity's Board of Directors or advisory committees. Martin:Abgenomics: Research Funding; Enlivex Therapeutics: Consultancy; Genentech: Consultancy, Other: One-time advisory board; Neovii: Other: One-time advisory board; Pfizer: Other: Data and Safety Monitoring Committee; Pharmacyclics LLC: Other: One-time advisory board; Procter and Gamble: Equity Ownership; Xenikos: Research Funding.
Novel Conditioning Regimen for Haploidentical Hematopoietic Cell Transplant for Severe Aplastic Anemia in Children
Authors: Hashem Hasan , Rihani Rawad , Khattab Eman , Abu Shanap Mayada , Tbakhi Abdelghani , Sultan Iyad 
Published: November 2019

New hematopoietic cell transplant (HCT) approaches are urgently needed for patients with severe aplastic anemia (SAA) who lack an HLA-identical donor. Haploidentical HCT with post transplant cyclophosphamide (PTCy) represent a potential universal available option for almost all children with SAA.
We present a novel conditioning regimen for haploidentical HCT in children with SAA in a center where horse ATG is not available. Conditioning regimen consists of rabbit ATG 2.5 mg/kg/day from day -9 to -7, Fludarabine 30 mg/kg/day from day -6 to -2, Cyclophosphamide 14.5 mg/kg/day from day -6 to -5, Thiotepa 5 mg/kg/day from day -4 to -3, and 4 Gy TBI on day -1 in in two fractions. GvHD prophylaxis consist of PTCy 50 mg/kg/day on days +3 and +4 along with Cyclosporine A and Mycophenolate mofetil (MMF) starting on day +5.
Four consecutive children with SAA referred to our center for haploidentical HCT starting in 2018. Median age at HCT was 9 years (5-16) with 3 males and 1 female. All patients were heavily transfused with both blood and platelets prior to referral for HCT. Two patients had strong and one had weak positive anti-HLA antibodies (DSAs) and received desensitization with IVIG, Rituximab and plasmapheresis. One patient received buffy coat infusion on day -1 due to persistent strong DSAs despite desensitization. Median CD34+ dose received was 12 x 10e6, and median CD3+ dose was 29 x 10e6. Donors were all same blood group to patients.
All patients successfully engrafted neutrophils at median of 13 days (12-14). Platelets engraftment in 3/4 patients at median of 7 days (5-10). All patients received peripheral blood as stem cell source. Three of four patients survived and doing well at last follow up. One patient had toxic death on day +38 due to chemotherapy related toxicity causing multi-organ failure. Chimerism analysis was full donor in all four patients at median follow up time of 11 months (2-12). Patients were sent home at median of 24 days post HCT. None developed grade 2-4 acute GvHD nor chronic GvHD. Acute GvHD of skin grade 1 stage 1 developed in 2 patient and managed with topical steroids. Viral reactivations consisted of CMV viremia and BK hemorrhagic cystitis in all patients, and have all resolved. No post transplant autoimmune complications.
Haploidentical HCT with PTCy represents a quick and first line approach in heavily transfused children with SAA. Although yet limited number of patients, this regimen is feasible and appears to be safe. A great advantage of this regimen is the rapid engraftment of both neutrophils and platelets. Moreover, although using peripheral blood as a stem cell source, there was no severe acute or chronic GvHD.

Disclosures
No relevant conflicts of interest to declare.