bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of Newborns Screening Laboratory Tests for Sickle Cell Disease and Other Haemoglobinopathies in Tanzania
Authors: Christopher Heavenlight Hebron , Burns Adam , Josephat Emmanuel , Saul Sephord , Mgaya Josephine , Makani Julie , Nkya Siana Watoky , Schuh Anna 
Published: November 2019

Affiliation
1Muhimbili University of Health and Allied Sciences, Dar es salaam, Tanzania.
2Department of Oncology, University of Oxford, Oxford UK.
3Dar es salaam University College of Education, Dar es salaam, Tanzania.
ABSTRACT.
Background; SCD constitutes to be a major public health problem in Tanzania. NBS for SCD identify infants with SCD at birth and subsequently enroll them into SCD comprehensive care program and it has been reported to reduce early childhood mortality and morbidity due to SCD and increase life expectancy. There are different methods for Newborn screening for SCD such as HPLC, IEF, DNA analysis, sickle scan and hemotype SC that helps in early detection of SCD and other haemoglobinopathies but we are lacking sufficient information on alternative screening methods that can be used for NBS in Tanzania so this study is aiming at investigating the effectiveness and feasibility of NBS laboratory tests.
Aim of the study; To evaluate newborn screening Laboratory assays for sickle cell disease and other haemoglobinopathies in Tanzania.
Methodology; This will be a retrospective and prospective study which will be conducted in Dar es Salaam and Mwanza.1000 newborns and 100 children with SCD under five years old will be enrolled. DBS samples will be collected and analyzed by DNA Analysis, sickle scan, Hemotype SC, HPLC, and IEF. SPSS version 16.0 will be used to analyze data. Statistically, significant tests will be declared at a level of p-value < 0.05.
Progress: Initially we have conducted a pilot of 12 individuals who were screened for SCD and turned out to be carriers ( Hb AS). However these individuals continuously reported to experience painful events. Initial analysis has include Hb subtyping and quantification by HPLC,Sickle scan and HemotypeSC to ascertain indicator for thalassaemia syndrome. Definitive analysis involved sequencing of thalassaemia and SCD associated regions by using Oxford Nanopore Technology and NGS
Expected outcomes; The findings of this study will inform Newborn screening stakeholders on the best point of care screening test and definitive test, in terms of turnaround time and cost. Such findings will assist in planning and scaling up of newborn screening services across the country.
Corresponding Author; HChristopher, Email; hchristopher@blood.ac.tz, Tel; +255762175732.

Disclosures
Schuh: AbbVie: Consultancy, Speakers Bureau; Genentech: Consultancy, Speakers Bureau; Pharmacyclics: Consultancy, Speakers Bureau; Kite: Speakers Bureau; Gilead: Speakers Bureau; Seattle Genetics: Speakers Bureau; Jazz Pharmaceuticals: Speakers Bureau; Bristol-Myers Squibb: Research Funding; Janssen: Speakers Bureau; Verastem: Speakers Bureau.
Preliminary Results of a Prospective Observational Study to Assess the Prevalence of Gaucher Disease in an Adult Population Affected By MGUS
Authors: Giuffrida Gaetano , Conticello Concetta , Nicolosi Daniela , Calafiore Valeria , Romano Alessandra , Consoli Ugo , Musolino Caterina , Innao Vanessa , Fiorenza Flavia , Maugeri Cinzia , Longo Giuseppe , Cingari Maria Rocca , Di Raimondo Francesco 
Published: November 2019

INTRODUCTION: Gaucher disease (GD) is a rare, autosomal recessive genetic disorder. It is due to a deficiency of the lysosomal enzyme, glucocerebrosidase, which leads to an accumulation of its substrate, glucosylceramide, in tissue macrophage with damage to hematological, visceral, and skeletal organ systems.
Although GD has a continuous spectrum of severity, it is traditionally classified into three forms: type 1 (chronic; lacking early onset neuronopathy), type 2 (acute; with early onset neuronopathy),and type 3 (chronic; with early onsetneuronopathy).
Type 1GD accounts for more than 90% all GD patients. Its prevalence world-wide is 1 in 50,000-100,000 but it is as high as ~1 in 850 in individuals of Ashkenazi heritage.
Type 1 GD is frequently associated to monoclonal gammopathies; despite the emergence of theories advanced to explain these observations, the cause remains unknown.
OBJECTIVE: Aim of the ongoing observational study is to determine the prevalence of unrecognized type I GD in a selected Italian population with MGUS.
MATERIALS AND METHODS: From January 2018, dried blood spots (DBS) sample from patients with laboratory evidence of MGUS coming from five hematology units of Sicily and Calabria were collected and tested for the acid β-glucosidase enzyme activity.
The study was approved by the local institutional review board. All patients provided informed consent for the prospective collection of their data.
In case of DBS positive result, a confirmatory test was carried over and, if GD was confirmed, the patient was referred to one of the Regional Reference Centers for Metabolic Disease, as for current clinical practice in Italy.
RESULTS: To date, 308 patients with MGUS were enrolled; acid β-glucosidase enzyme activity was low in 22 patients (7%). Sequence analysis of GBA gene was performed in these selected patients, but we have found only 4 patients with heterozygous mutation in the GBA1 gene, 1 homozygous(c.1226A>G -N370S) and 1 compound heterozygous (c.1226A>G -N370S and c.1448T>C -L444P); the last 2 patients had signs of GD (hepato-splenomegaly and mild thrombocytopenia).
CONCLUSIONS: Type 1 GD remains a rare lysosomal storage disorder but preliminary results of our observational study show that it should be considered in the diagnostic framework of patients with MGUS, particularly when other GD symptoms are present.

Disclosures
Conticello: Celgene: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding. Di Raimondo:Celgene: Consultancy, Honoraria, Research Funding; Takeda: Consultancy; Amgen: Consultancy, Honoraria, Research Funding.
Venetoclax Is Synergistic with Idelalisib or MK2206 Against Primary CLL Cells in an in Vitro Model of the Microenvironment
Authors: Shen Yandong , Crassini Kyle R , Fatima Narjis , Christopherson Richard , Mulligan Stephen P. , Best Oliver Giles 
Published: November 2019

Background
The PI3-kinase signaling pathway and the Bcl-2-family of proteins play crucial roles in regulating the survival and proliferation of chronic lymphocytic leukemia (CLL) cells in the bone marrow and lymph nodes. Trials of ibrutinib, idelalisib and venetoclax illustrate the potential of targeting the B-cell receptor (BCR) signaling pathway and Bcl-2, however disease relapse is still common. Several pre-clinical studies and on-going clinical trials [Rogers et al., 2018, Jain et al., 2019] suggest that combinations of BCR inhibitors with venetoclax may be an effective treatment strategy for CLL patients with high risk disease. We sought to investigate the effects of combining idelalisib or the AKT inhibitor MK2206 with venetoclax against CLL cells under in vitro conditions that mimic the tumor microenvironment.
Methods
Primary CLL cells were co-cultured with CD40L-expressing mouse L-fibroblasts. Cell viability was assessed using the mitochondrial membrane potential dye DilC1(5), propidium iodide and flow cytometry (n = 6). Synergy between idelalisib or MK2206 and venetoclax was evaluated by calculating combination indices (CI) using the Compusyn software. The mechanisms of action of the drugs and synergies between the drugs were investigated by immunoblotting (n = 6).
Results
Venetoclax was highly synergistic in combination with idelalisib or MK2206 against CLL cells co-cultured with CD40L-fibroblasts, with CI values of 0.2 and 0.5 at a fractional effect of 0.9, respectively (Figure 1). This synergy was consistent with a significant (P < 0.05) reduction in the IC50 for venetoclax, idelalisib and MK2206.
Immunoblotting suggests that MK2206, as a single agent or in combination with venetoclax, was more effective than idelalisib in inhibiting the phosphorylation of AKT and NF-κB. Both MK2206 and idelalisib as single agents and in combination with venetoclax significantly reduced expression of Mcl-1 and Bfl-1, two pro-survival members of the Bcl-2 family of proteins in primary CLL cells co-cultured with CD40L-fibroblasts.
Conclusions
The synergy observed, which was associated with a significant decrease in the IC50s for idelalisib and MK2206, may mitigate some of the toxicities associated with PI3-kinase pathway inhibitors. Comparison of the two PI3-kinase-pathway inhibitors suggests that MK2206 may be more effective than idelalisib at blocking BCR-mediated signaling as a single agent and in combination with venetoclax. The mechanisms underlying the synergy include down-regulation of expression of Bcl-2 family proteins that are not targeted by venetoclax as a single agent. The data presented support the rationale for on-going and future clinical trials of combination therapies incorporating a PI3-kinase inhibitor with venetoclax for the treatment of high risk CLL.
Figure 1

Disclosures
No relevant conflicts of interest to declare.
Population Level Outcomes of Chronic Lymphocytic Leukemia in the Era of Targeted Agents- Analysis of Surveillance Epidemiology and End Results (SEER) Database
Authors: Narra Ravi Kishore , Atallah Ehab L. , Hamadani Mehdi , Guru Murthy Guru Subramanian Guru 
Published: November 2019

Background:
The treatment paradigm of chronic lymphocytic leukemia (CLL) has changed significantly in the last decade with approval of multiple new targeted agents. Small molecule drugs such as ibrutinib, venetoclax, idelalisib and other targeted agents have improved the outcomes of patients with CLL treated in clinical trials. However, it is unclear if the population level outcomes of CLL have improved since the first approval of bruton tyrosine kinase (BTK) inhibitor Ibrutinib in February,2014.
Methods: Using SEER database, we identified patients aged ≥ 20 years with pathologically confirmed CLL (ICD-0-3 code 9823/3) diagnosed between 2011-2016 and actively followed. Patients were divided into two groups by the period of diagnosis - 2011-2013, 2014-2016, reflecting the period pre-and post-ibrutinib approval. Overall survival (OS) was compared between the two groups using Kaplan-Meier method and log rank test. Cox proportional hazard regression method was used to determine the influence of period and demographic factors on OS. Statistical analysis was performed with significant two-sided p< 0.05.
Results: A total of 30701 patients with a median age of 69 years (range 21-104) were included. Males (60.9%) and White race (80.8%) contributed to the majority as summarized in table 1. Median OS was not reached. OS at 30 months was significantly higher for patients diagnosed from 2014-2016 (83.3%) compared to those diagnosed from 2011-2013 (81.7%) (p=0.004) (figure 1). On multivariate analysis, diagnosis from 2014-2016 (HR 0.90, 95% CI 0.85-0.96, p: 0.002) and females (HR 0.74, 95% CI 0.70-0.78, p< 0.001) had lower mortality risk, whereas, increasing age (HR 1.07, 95% CI 1.07-1.08, p< 0.001), Hispanics (HR 1.18, 95% CI 1.05-1.32, p< 0.004), Non-Hispanic blacks (HR 1.41, 95% CI 1.28-1.54, p< 0.001) had higher risk of mortality.
Conclusions: Survival of patients with CLL at the population level is improving since approval of Ibrutinib. Non-biological factors such as age, gender and race/ethnicity continue to influence the survival even in the era of novel agents, highlighting the need for continued research to address these discrepancies.

Disclosures
Atallah: Jazz: Consultancy; Jazz: Consultancy; Novartis: Consultancy; Pfizer: Consultancy; Helsinn: Consultancy; Helsinn: Consultancy; Takeda: Consultancy, Research Funding. Hamadani:Janssen: Consultancy; Merck: Research Funding; ADC Therapeutics: Consultancy, Research Funding; Pharmacyclics: Consultancy; Takeda: Research Funding; Celgene: Consultancy; Sanofi Genzyme: Research Funding, Speakers Bureau; Medimmune: Consultancy, Research Funding; Otsuka: Research Funding. Guru Murthy:Cardinal Health Inc.: Honoraria.
Sarcopenia As a Significant Prognostic Factor in Acute Myeloid Leukemia: Validation of a Novel Scoring System
Authors: Sun Qian , Ming Hong , Qian Sixuan , Li Jian-Yong 
Published: November 2019

Background: Acute myeloid leukemia (AML) is a malignant hematologic disease mainly occurs in the elderly. Sarcopenia is a muscle disease (muscle failure) common among adults of older age but can also occur earlier in life. This study indicates to investigate the prognostic value of sarcopenia in AML patients.
Methods: From January 2014 to April 2019, a total of 100 patients diagnosed with de novo AML according to the WHO classification were included in this study. Clinical data and biological characteristics were collected. Giemsa R-banding method was used to detect the karyotype of metaphase bone marrow cells. Targeted gene sequencing was taken to test a total of 23 gene mutations including DNMT3A, IDH1, IDH2, ASXL1, RUNX1, NPM1, CEBPA, FLT3, KIT, and TP53, etc. Bioelectrical impedance analysis (BIA) of human body before treatment was used to calculate the Body Mass Index (BMI), Appendicular Skeletal Muscle Mass (ASM), SMI (ASM/height2), Fat Mass (FM), Fat-free Mass (FFM), and FFMI (FFM/height2), etc. The diagnosis of sarcopenia was based on the consensus report of the Asian Working Group for Sarcopenia.
Results: Sixteen AML patients were diagnosed as sarcopenia. Patients with sarcopenia had significantly lower weight (P=0.049), BMI (P=0.039), ASM (P=0.002), SMI (P<0.001) and FFMI (P=0.005) than non-sarcopenia ones (Figure 1A~E). With a median follow-up time of 25 months, the overall survival (OS) of patients with sarcopenia was shorter than non-sarcopenia patients (median 7 vs. 28 months, P=0.054, Figure 2A). When stratified by age (≥60 yrs vs. <60 yrs), elderly patients with sarcopenia showed a significantly inferior OS to normal elder ones (median 3 vs. 12 months, P=0.008, Figure 2B). Receiver operator characteristic curves suggested that sarcopenia as one additional point to the 2017 European Leukemia Net risk stratification by genetics obtained superior area under the curve (AUC) in predicting AML OS than the latter alone (AUC 0.67 vs. 0.649, P=0.3305, Kaplan-Meier plotting see Figure 2C~D).
Conclusion: Sarcopenia is common in AML patients, and BIA offers a useful objective tool for identifying these patients. We illustrated a marked clinical impact of sarcopenia on patients with AML, and presented a novel prognostic scoring system. In elderly patients, the presence of sarcopenia before treatment may offer a useful factor for avoiding intensive therapeutic strategies for AML.

Disclosures
No relevant conflicts of interest to declare.
ATG4A is a Novel Regulator of Mitophagy during Human Erythropoiesis
Authors: Stolla Massiel Chavez , Thom Neele , Reilly Andreea , Clough Courtnee , Abkowitz Janis L. , Doulatov Sergei 
Published: November 2019

Autophagy is a highly conserved pathway that degrades and recycles intracellular components. While autophagy is activated in response to cellular stress, it also contributes to hematopoietic cell differentiation. Previous studies in mice lacking core autophagy proteins have identified mitochondrial clearance as a critical function of autophagy in erythroid differentiation and maturation. Impaired autophagy results in anemia, retained mitochondria and elevated levels of reactive oxygen species in mice. However, the kinetics and regulation of mitophagy in human erythropoiesis have not been investigated. To better understand the kinetics of mitophagy in human erythropoiesis we have developed a lentiviral mitophagy reporter (MT-Keima) and used it to monitor mitochondrial clearance during erythroid differentiation of primary human CD34+ hematopoietic stem and progenitor cells (HSPCs). Measurement of mitophagy during erythroid differentiation revealed active mitochondrial clearance during early and late stages of erythropoiesis culminating with total clearance of mitochondria at the terminal stages of differentiation. Gene expression analysis during human erythroid differentiation identified the upregulation of the core autophagy program including ATG4A encoding a cysteine protease, which was restricted to the erythroid lineage. Knockdown of ATG4A in primary human HSPCs significantly reduced mitophagy in early and late erythroid cells and resulted in increased mitochondrial mass in terminally differentiated reticulocytes. Furthermore decreased expression of ATG4A, but not its paralog ATG4B, reduced the (i) expansion of erythroid progenitors, (ii) total number of erythroblasts (iii) and significantly reduced enucleation relative to luciferase controls. Finally significantly fewer erythroid colonies were found in methylcellulose culture only in HSPCs with ATG4A KD when compared to luciferase controls, while numbers of myeloid colonies were preserved, supporting a role for ATG4A selectively in the human erythroid lineage. Together these results identify ATG4A as a novel erythroid-specific regulator of mitophagy and a new potential target for the therapeutic modulation of autophagy in human erythropoiesis.

Disclosures
No relevant conflicts of interest to declare.
Impact of ABO Blood Group Incompatibility on Outcomes after Single-Unit Umbilical Cord Blood Transplantation for Malignant Hematological Disease
Authors: Siqi Cheng , Tang Baolin , Zhu Xiaoyu , Liu Huilan , Song Kaidi , Wan Xiang , Yao Wen , Wang Jian , Sun Zimin 
Published: November 2019

Objective
In contrast to solid organ transplantation, ABO blood group incompatibility was acceptable in allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, reports of the effect of donor-recipient ABO incompatibility on long-time survival, graft-versus-host disease (GVHD), and relapse after allo-HSCT were controversial. Relatively few reports existed on the effects of ABO incompatibility after umbilical cord blood transplantation (UCBT). The aim of this study was to investigate the role of major ABO incompatibility on RBC transfusion burden, hematologic recovery, GVHD, transplant-related mortality (TRM), relapse, and overall survival (OS) in UCBT for malignant disease.
Methods
This retrospective study included 587 malignant hematonosis patients who received myeloablative single-unit unrelated donor UCBT at our center between May 2008 and June 2018. Median follow-up time of the patients alive was 40.7 months (range: 12.0-134.6 months). A total of 230 (39.2%) patients received an ABO-identical transplant, and 357 (60.8%) received ABO-mismatched transplants, including 161 (27.4%) minor, 141 (24.0%) major, and 55 (9.4%) bidirectional ABO-incompatible UCBTs. All patients received myeloablative conditioning regimens and cyclosporine A (CsA) combined with mycophenolate mofetil (MMF) as a GVHD prophylaxis.
Results
A comparison of ABO compatibility and incompatibility demonstrated no significant differences (P>0.05) in the cumulative incidence of neutrophil, platelet, and red blood cell engraftment . There was no significant difference in the cumulative incidence of grades Ⅱ to Ⅳ aGVHD (P= .527) and Ⅲ to Ⅳ aGVHD (P= .949) among the 4 groups (Figure A , B). In univariate analysis, ABO blood group incompatibility was not associated with cumulative incidence of 180d TRM (Figure C, P= .602). The overall 3-year survival had no statistically significant differences among the 4 groups (Figure D; P= .384). Further, 11 patients were excluded from the analysis of post-UCBT RBC transfusion burden because of missing data and non-red blood cell engraftment. Of the remaining 576 patients, the median number of RBC transfusions during transplant days 0 to 60 was 4 (range, 0 to 106). There was no significant difference in the transfusion burden among all ABO blood type mismatch groups (Table 1, P = .069). Furthermore, none of the patients developed pure red aplastic anemia (PRCA) after UCBT.
Conclusion
The results showed that ABO blood group incompatibility had no significant impact on hematologic engraftment, the occurrence of GVHD, and the survival of malignant hemoblastosis. Patients with myeloablative single-unit UCBT may not develop PRCA; Donor-recipient ABO incompatibility may not be the major consideration in the selection of umbilical cord blood.

Disclosures
No relevant conflicts of interest to declare.
May Tigit (T cell Ig and ITIM domain) Expression be a New Target for Hodgkin Lymphoma?
Authors: Annibali Ombretta , Bianchi Antonella , Grifoni Alba , Tomarchio Valeria , Tafuri Mariantonietta , Verri Martina , Avvisati Giuseppe , Crescenzi Anna 
Published: November 2019

Introduction: The use of immune-checkpoints (anti PD-1/PD-L1) resulted very effective in the treatment of relapsed/refractory classical Hodgkin Lymphoma (cHL). T cell Ig and ITIM domains (TIGIT) is another immune checkpoint receptor known to negatively regulate T cell functions.
Methods: In this study, we investigated the expression of TIGIT in cHL microenvironment in order to find a potential new target for inhibitor therapy. TIGIT, PD-1 and PD-L1 expression was evaluated in 34 consecutive patients with cHL referred to Campus Bio-Medico University Hospital for diagnosis and treatment during the last eight years.
Results: TIGIT positivity was observed in 19/34 patients (56%), of these 11 (58%) had advanced stages and 10 (52%) B symptoms. Among the 8 patients with IPS ≥ 4, 6 (75%) were TIGIT +.
Out of the 19 TIGIT + (score 1 to 3) peritumoral T lymphocytes, 16 (84%) were also PD-1+. While out of 15 TIGIT- peritumoral T lymphocytes only 4 (27%) were PD-1+ and 11 (73%) were PD1- (P=0.001). Moreover all 15 TIGIT- patients were PD-L1+ (100%) while out of 19 TIGIT + patients, 11 (58%) were PD-L1+ and 8 (42%) PD-L1- (P=0.004).
Conclusion: This study indicate that the great majority of TIGIT+ patients are also PD-1+, while those who are TIGIT- are all PD-L1+. These results may offer a new therapeutic possibility in the treatment of Relapsed/Refractory (R/R) cHL by the use of anti-TIGIT monoclonal antibodies.
Figure