bluuebunny/crossref_metadata_2025 huggingface dataset:
A Meta-Analysis of Genetic Abnormalities and Next Generation Sequencing of Primary Cases of Castleman Disease
Authors: Butzmann Alexandra , Przybyl Joanna , Sridhar Kaushik , Kumar Jyoti , Brar Nivaz , Zehnder James L , Warnke Roger A , Ohgami Robert S.
Published: November 2019

Castleman disease is a rare lymphoproliferative disorder known to represent at least four distinct clinico-pathologic sub-types. Large advancements in our clinical and histopathologic description of these diverse diseases have been made, resulting in subtyping based on degree of nodal involvement (unicentric versus multicentric) and according to viral infection (human herpes virus 8 - HHV8 and Human Immunodeficiency Virus - HIV). However, we continue to lack a foundational understanding of the biologic mechanisms driving at least a subset of patients with these diseases. In recent years, significant molecular and genetic abnormalities that may be associated with this disease have been described; yet we are still unclear as to a unified vision of what loci and genes are involved in at least a subset of cases of Castleman disease.

In our study we performed a meta-analysis of all cases (unicentric and multicentric) of Castleman disease described in literature to date, and correlate cytogenetic, molecular and genetic abnormalities with disease subtypes, phenotypes and clinical outcomes. We further performed targeted DNA deep sequencing and RNA-sequencing on a cohort of Castleman Disease cases.

Our results demonstrate that genetic abnormalities in the MAPK-pathways, PI3K-pathway and Interleukin signaling are frequent in unicentric Castleman disease, whereas abnormalities in DNA methylation are seen more commonly in idiopathic multicentric Castleman disease, along with abnormalities in MAPK-pathways.

We conclude that UCD and iMCD may share similar pathways of MAPK pathogenesis but can also be distinguished by cytokine and other DNA modification pathways that differ.

Disclosures

Ohgami: Agilent technologies: Other: received support/funding.
Mitigation of Venous Thromboembolism Risk through Favorable Lifestyle: The ARIC Study
Authors: Evans Christina , Hong Ching-Png , Folsom Aaron R , Heckbert Susan , Smith Nicholas , Wiggins Kerri , Cushman Mary
Published: November 2019

Background: Venous thromboembolism (VTE) is a common disease with a strong genetic basis. Unhealthy lifestyle factors contribute to risk, but it is unknown whether healthier lifestyle can mitigate the risk for VTE in those at high genetic risk. We studied whether greater adherence to the American Heart Association's (AHA's) cardiovascular health metric called Life's Simple 7 (LS7) is associated with a lower rate of VTE in individuals with high genetic risk score (GRS) for VTE.

Methods: We followed 9,026 middle-aged white participants from the Atherosclerosis Risk in Communities (ARIC) Study, a prospective cohort of 15,792 individuals enrolled in 1987-89. A validated GRS was used, comprising 5 well known genetic conditions associated with VTE (factor V Leiden, prothrombin 20210A, non-O blood group, factor XI rs4241824, and fibrinogen gamma FGG rs2066865). Only white participants were included, as the GRS did not predict VTE in others. AHA's LS7 categories of inadequate, average, and optimal health were determined based on smoking status, body mass index, physical activity, diet, total cholesterol, blood pressure, and fasting glucose. VTE events were adjudicated by expert medical record review. We calculated hazard ratios (HRs) and 95% confidence intervals (CI) of incident VTE by LS7 categories, stratified by GRS (low, intermediate, high), adjusting for age, sex, and education. HRs were also calculated for individual LS7 components stratified by GRS.

Results: There were 466 incident VTE over 22.8 years of follow-up. Compared to those with optimal health, those with inadequate LS7 score had higher rates of VTE (5.7% vs. 3.9%). In Figure 1, compared to the high GRS / inadequate LS7 group, the HR of VTE in the low GRS group with optimal health was lowest at 0.39 (95% CI 0.25-0.61), but moreover, the HR in the high GRS group with optimal health was also attenuated to 0.65 (95% CI 0.48-0.89). The pattern of association was similar for provoked and unprovoked VTE. Of the LS7 components, obesity was most strongly related to VTE. In Figure 2, compared to obese / high GRS participants, the HR of VTE with normal weight / low GRS was 0.36 (95% CI 0.23-0.57), while the HR in high GRS / normal weight participants was reduced by 45%, at 0.55 (95% CI 0.4-0.76).

Conclusion: Among all participants, even those at high genetic risk, healthier lifestyle factors, particularly obesity, were associated with decreased incidence of VTE. Further studies should determine the impact of lifestyle change among patients at high genetic risk of VTE, such as in thrombophilic families.

Disclosures

Heckbert: National Institutes of Health: Other: Grants.
Tiered Oral Therapy Protocol for Sickle Cell Vaso-Occlusive Crisis
Authors: Guy Margaret , Magee Lauren , Keiser Jessica , Carter Nicole , Vaughan DaleMarie , McHenry Chantal , Sop Daniel , Hartigan Sarah , Smith Wally R
Published: November 2019

Background: Adults with sickle cell disease (SCD) often present with extremely painful acute vaso-occlusive crises (VOC). VOC guidelines quote studies supporting early initiation of opioids, but are lacking in a standardized approach to achieve adequate analgesia and subsequent de-escalation. Various SCD studies comparing continuous basal versus demand bolus patient-controlled analgesia (PCA) show conflicting outcomes. Thus the objective of our pilot was to develop a standardized approach to pain management utilizing PCA and a Tiered Oral Therapy Protocol (TOTP) in an effort to affect healthcare utilization by decreasing length of stay without increasing readmission rates.

Methods: We studied admissions of 31 SCD (any genotype) patients (8 males, 23 females) ≥ 18 years old, all African-American, admitted for VOC, and treated by hospitalist providers using TOTP on one medical unit at Virginia Commonwealth University Health System from July 2018 to June 2019. Inpatient admissions for one calendar year prior to the initial TOTP admission served as the patient's historical control. TOTP is guided by the patient's individualized treatment plan and starts with continuation of their home basal dosing, demand dose only PCA, and immediate-release oral opioids for additional breakthrough pain. TOTP goals are aggressive up-titration of opioids until adequate analgesia. After a period of stability the next phase encourages timely, aggressive down-titration of opioids to speed discharge. We primarily compared length of stay (LOS), hospital charges, and 30-day readmission rates for TOTP vs historical controls. Secondarily, we compared adverse safety events: opioid-related rapid responses, naloxone administration, or falls.

Results: Utilization of TOTP resulted in reductions in LOS (21%, 4.7 days controls vs. 3.7 days TOTP, p<0.014), and hospital charges ($1,627,497.73 controls vs. $944,528.96 TOTP, p=0.1403), and an insignificant increase in the 30-day readmission rate (35.6% controls vs. 42.7% TOTP, p=0.5027). Hospital charge data is difficult to interpret as a full year of post TOTP encounters are not available for all patients. There were no TOTP vs. historical control differences in opioid-related safety events.

Conclusion: Standardization of therapy using the TOTP improved length of stay and reduced charges for patients with sickle cell disease admitted for acute VOC without increasing adverse safety events or significantly impacting the 30 day readmission rate. We have enlarged this pilot to include other VCU medical units, to test whether we can reproduce our initial one-unit success. Further, we recommend TOTP implementation and testing in other hospitals to exclude secondary trends, since TOTP was but one among several interventions in a VCU Adult SCD Medical Home.

Figure

Disclosures

Smith: Novartis: Consultancy, Honoraria.
Mutation of Murine Rpl5 reveals a New Model for Diamond Blackfan Anemia Characterized By Defective Erythropoiesis
Authors: Lemay Philippe , Jones Morgan A , Mohamed Fatma , Guyot Marie-Claude , Ludlow Alexander , Li Qing , Engel James Douglas , Franceschi Renny , Rothstein Thomas L. , Kaartinen Vesa , Kibar Zoha , Singh Sharon A
Published: November 2019

Background

Ribosome dysfunction is implicated in several abnormal developmental conditions and diseases in humans. Heterozygous germline ribosomal protein (RP) variants are found in the majority of individuals with Diamond Blackfan anemia (DBA) with RPL5 (uL18) being the second most frequently mutated gene. Patients with RPL5 mutations are for unknown reasons at increased risk of congenital defects when compared to other genotypes. There is currently no available Rpl5 mouse model with anemia and therefore hematopoiesis in this genotype has not been well studied.

Methods

We characterized a novel mutant mouse with a growth and kinky tail defect, and increased mortality. The mouse was generated in a chromosome 11 balancer ENU mutagenesis screen. Following genetic mapping and whole exome sequencing, we identified a heterozygous intronic Rpl5 missense variant, which segregated with all affected mice. We first performed molecular analyses to validate the impact of this variant on RPL5 and ribosome levels, and then examined the development and physiology of multiple tissues by histology, micro-CT, echocardiogram and measurement of erythroid differentiation to determine the etiology of the observed increased mortality in newborn Rpl5 mutant mice.

Results

This variant led to decreased Rpl5 mRNA and protein, and consequential polysome defects (decreased 60S and 80S ribosome subunits) consistent with RPL5 haploinsufficiency. All Rpl5+/- neonates exhibited deficient terminal erythroid differentiation. A fraction of the mutants progressed to severe anemia leading to increased mortality while others appeared to undergo spontaneous remission. Surviving adult/aged mice exhibited no anemia although preliminary studies indicate reduced numbers of hematopoietic progenitors. Most newborn mutant mice had large ventricular septal defects (VSD) while surviving adult mice had small VSDs on echocardiogram. Rpl5+/- mice were significantly smaller without craniofacial abnormalities as assessed by micro-CT or histology. Bone and cartilage staining of newborn mice revealed that the kinky tail defect resulted from delayed endochondral ossification.

Conclusion

This model recapitulates the major developmental and erythroid differentiation defects of DBA in an animal model. We propose that this novel Rpl5 mutant provides an opportunity to study the mechanism of the variable penetrance of anemia and the congenital defects normally encountered in DBA.

Disclosures

Singh: Emmaus Medical, Inc: Consultancy.
Platelet Phenotype Prediction from Whole Genome Sequencing in 621 Sickle Cell Disease Patients
Authors: Montemayor Garcia Celina , Weisman Julie K. , Wang Xunde , Gandla Divya , Long James Owen , Dalgard Clifton L. , Pirooznia Mehdi , Klein Harvey , Thein Swee Lay
Published: November 2019

Introduction: Patients with sickle cell disease (SCD) are at increased risk of alloimmunization. Platelet refractoriness is a serious known complication and often seen in SCD patients who are heavily transfused and/or in the bone marrow transplantation (BMT) setting. Next generation sequencing (NGS) is an emerging and promising genotyping strategy in the context of blood typing, due its high throughput and its ability to detect both known and novel variants in the patient and donor population. Here we describe an algorithm to predict common and rare human platelet antigens (HPA) from NGS data, and its validation through Sanger sequencing.

Design/Methods: Whole genome sequencing (WGS) was performed on stored blood samples from 621 SCD patients enrolled in 2 IRB-approved clinical studies. Our open source software RyLAN (Red Cell and Lymphocyte Antigen prediction from NGS) was utilized to translate WGS data into predicted RBC and platelet phenotypes. The 29 genomic variants interpreted by RyLAN in 6 HPA genes were correlated with Sanger sequencing.

Results: Our study cohort consisted of 621 SCD patients (485 HbSS, 21 HbSβ0, 29 HbSβ+, 84 HbSC, 1HbS O Arab, and 1 HbSD). The mean age was 34.3 years, and 46% were male. Previous red cell transfusions were recorded in 62% of patients, and 3% were documented as never transfused. RyLAN software was executed as a singularity container in multithreaded mode, completing analysis of all 621 .bam WGS files in 18 hours. RyLAN predicted 237 unique extended platelet phenotype combinations in this cohort, with an average read depth of 33 in genomic areas of interest. Predictions for 10 platelet antigens in 26 participants, including rare phenotypes like HPA-25bw+ and HPA-13bw+, were confirmed by bidirectional Sanger.

Conclusions: We describe an efficient, open-source algorithm used to interpret 6 HPA genes from WGS in a large SCD cohort. WGS, in conjunction with the RyLAN algorithm, demonstrated 100% accuracy in predicting common and rare HPA genomic variants. Future studies are needed to refine WGS algorithms in SCD, and to examine the possible value of this technology in HPA alloantibody identification workups, optimal platelet product allocation, and donor recruitment.

Disclosures

No relevant conflicts of interest to declare.
The ''Gatekeeper'' Mutation T315I in BCR/ABL Confers Additional Oncogenic Activities to Philadelphia Chromosome Positive Leukemia
Authors: Zafar Usva , Yusuf Mohammed , Chakraborty Rikhia , Lalani El-Nasir M A , Mian Afsar Ali
Published: November 2019

Chronic myeloid leukemia (CML) and 30% of adult acute lymphatic leukemia (ALL) are characterized by the Philadelphia chromosome (Ph+), having a (9;22) chromosomal translocation. The BCR/ABL fusion protein is the hallmark of Ph+ leukemia. BCR/ABL is characterized by deregulated and constitutively activated ABL tyrosine kinase activity that determines its transformation potential. Tyrosine kinase inhibitors (TKI) have greatly improved the overall prognosis of these diseases, particularly by altering the natural history of chronic phase (CP) CML and preventing the previously inexorable progression to terminal blast crisis (BC). However, unsatisfactory responses in advanced disease stages, resistance and long-term tolerability of BCR/ABL inhibitors represent major clinical problems. The most important mechanism of resistance against TKIs is the selection of leukemic clones driven by BCR/ABL harboring point mutations, such as the E255K, Y253F/H (P-loop), H396R (activation loop) or the T315I (gatekeeper). The "gatekeeper" mutation T315I confers resistance against all approved TKIs, with the only exception of Ponatinib, a multi-target kinase inhibitor. CML and Ph+ ALL, rarely present at diagnosis with a BCR/ABL harboring a resistance mutation to TKI. Resistant clones may be present and only detectable by highly sensitive methods. We have previously shown that the resistance mutations may influence the biology of BCR/ABL and its transformation potential.

We therefore hypothesized that the presence of mutations such as the T315I select for a "dormant cell population" which manifests following initial treatments with TKI inhibitors and treatment failure. The aim of this study was to determine whether the ''gatekeeper'' mutation T315I is able to confer biological features to BCR/ABL influencing its leukemogenic potential.

We investigated the influence of T315I on the biology of BCR/ABL in CML and Ph+ ALL. We used Ph+ ALL patient derived long term culture (PDLTCs), factor dependent Ba/F3 cells and syngeneic mouse model of BCR/ABL induced CML-like disease. These models allowed the direct comparison of BCR/ABL with BCR/ABL-T315I. We observed significantly slower proliferation of Ba/F3 cells and PDLTCs expressing BCR/ABL-T315I compared to the native BCR/ABL. This was further confirmed by undertaking mitotic index calculations and colony formation assays on both cell types. Furthermore, the induction of a CML-like disease in syngeneic mice was significantly delayed in the presence of T315I (median: BCR/ABL - 27 days; BCR/ABL-T315I - 61 days). We undertook functional studies to determine the putative signaling pathway and found that Ras/Erk1/2 pathway was activated inT315I positive cells.

This study may assist towards therapy decisions in patients with CML/Ph+ ALL with a T315I mutation.

Disclosures

No relevant conflicts of interest to declare.
Cost-Effectiveness of Avatrombopag for the Treatment of Thrombocytopenia in Patients with Chronic Liver Disease
Authors: Allen Lee F. , Aggarwal Kavita , Vredenburg Michael , Barnett Christine , Mladsi Deirdre , Kim Ray
Published: November 2019

Background: Patients with chronic liver disease (CLD) often have severe thrombocytopenia (platelet counts <50,000/mL) that can complicate the invasive diagnostic and therapeutic procedures these patients require as part of their clinical management, due to the increased bleeding risk. Avatrombopag is a thrombopoietin receptor agonist (TPO-RA) that is approved for the treatment of thrombocytopenia in patients with CLD as an alternative to platelet transfusions for patients undergoing a procedure. The aim of this study was to evaluate the relative cost-effectiveness of avatrombopag compared with platelet transfusion or treatment with lusutrombopag, another TPO-RA also approved for the treatment of thrombocytopenia in adult patients with CLD.

Methods: A decision-tree model was developed from a US payer perspective to capture the clinical events observed in registration trials, and to project potential longer-term complications resulting from a major bleed or thromboembolic event in the scenario analyses. Treatment costs were taken from publicly available data sources; avatrombopag and lusutrombopag estimates were calculated from the US prescribing information and Phase 3 study data. The interventions were evaluated in the overall trial population, patients with platelet counts <50,000/mL, and in subpopulations with higher (>40,000/mL to <50,000/mL) and lower (<40,000/mL) Baseline platelet counts. The primary metric for this economic analysis was the per-person total cost, the cost of prophylactic platelet transfusions required, and the incremental cost per prophylactic platelet transfusion avoided.

Results: In the overall population, avatrombopag reduced the need for platelet transfusions and produced cost-savings per person compared to Intended Platelet Transfusion (80% fewer prophylactic platelet transfusions), resulting in a relative cost savings of $4,250. The cost for lusutrombopag (15% more platelet transfusions) relative to avatrombopag was $5,819 higher than the cost of avatrombopag. Similar results were seen in both the higher and lower platelet count subgroups. The one-way and probabilistic sensitivity analyses showed that the use of avatrombopag remained cost-saving over a wide range of changes in input variables, with the incremental cost-effectiveness ratio falling into quadrant IV (decreased costs while prophylactic platelet transfusions were avoided).

Conclusions: From the cost-effectiveness standpoint, the use of avatrombopag is a practical strategy compared with the cost of both platelet transfusion and lusutrombopag, as it saves costs and reduces the need for prophylactic platelet transfusions.

Table

Disclosures

Allen: Dova Pharmaceuticals: Equity Ownership, Other: Chief Medical Officer . Aggarwal:Dova Pharmaceuticals: Employment. Vredenburg:Dova Pharmaceuticals: Employment, Other: Shareholder.
Preemptive Therapy Based on the Load and Time of EBV-Emia for Preventing EBV Related PTLD after Haploidentical Hematopoietic Stem Cell Transplantation: A Prospective Multicenter Study
Authors: Liu Can , Lin Ren , Wang Yu , Huang Fen , Fan Zhiping , Zhang Shen , Xu Yajing , Yang Ting , Xuan Li , Xu Na , Ye Jieyu , Sun Jing , Huang Xiaojun , Liu Qifa
Published: November 2019

Backgroud: To evaluate the efficacy of preemptive therapy for preventing EBV related PTLD based on the duration of EBV-emia and the viral loads in the patients undergoing haplo-HSCT, a prospective multicenter study was conducted.

Methods: Rituximab or rituximab followed by adoptive cellular therapy was applied for preemptive therapy if EBV-DNA in blood was positive twice consecutively with a rising trend over one log in virus loads or positive consecutively six times without a dropping trend over one log at least within three weeks or not turning negative consecutively eight times within four weeks.

Results : Of the 408 patients with acute leukemia enrolled in this study, 133 occurred EBV-emia, including 19 PTLD and 6 end-organ diseases. The patients with EBV-emia had inferior three-year overall survival (OS) and higher non-relapse mortality (NRM) to those without EBV-emia (OS: 58.0%±5.0% vs. 71.0%±3.0%, =0.042; NRM: 27.9%±3.9% vs. 18.9%±2.4%, =0.050, respectively). Compared with non-preemptive group, the negative conversion of EBV-emia was significantly higher (100% vs. 76.1%, =0.002) and the three-year incidence of PTLD was markedly lower in preemptive group (1.3% vs. 21.7%, =0.000). The median time from EBV-emia to EBV diseases onset was 15 days and the durative time of EBV-emia was positively correlated with PTLD occurrence (=0.419, =0.024).

Conclusion s: Rituximab preemptive therapy could reduce the incidence of PTLD. Earlier preemptive therapy should be advocated once the EBV-emia is positive persistently regardless of EBV-DNA levels (NCT01883180).

Disclosures

No relevant conflicts of interest to declare.
Outcomes of Primary and Secondary Prophylaxis of Chemotherapy Induced and Febrile Neutropenia (CIN/FN) in Bendamustine Plus Rituximab (BR) Regimens in Patients with Lymphoma and Chronic Lymphocytic Leukemia (CLL): Real-World, Single-Center Experience
Authors: Moore Logan , Bartels Trace , Persky Daniel O. , Kumar Abhijeet , Abraham Ivo , Sanders Lauren , Ortega Alicia , Henglefelt Alyssa , McBride Ali
Published: November 2019

Introduction:

Granulocyte stimulating growth factors (G-CSF) such as filgrastim or pegfilgrastim are indicated as prophylaxis of chemotherapy-induced neutropenia (CIN) and febrile neutropenia (FN). BR regimen is considered an intermediate FN risk (10-20%) per National Comprehensive Cancer Network guidelines. Therefore, patients receiving BR need to be assessed using patient specific risk factors to evaluate the need for primary prophylaxis. This study evaluates real-world patterns and outcomes associated with primary and secondary G-CSF prophylaxis in patients with B-cell lymphoma and CLL treated with BR.

Methods:

Retrospective chart review of all lymphoma or CLL patients treated with BR from 11/2013 through 6/2019 at the University of Arizona Cancer Center was conducted. Baseline demographic and chemotherapy cycle data was analyzed through Chi-Squared test and Unpaired t-test with a-priori p-value of 0.05 being considered statistically significant.

Results:

Eighty-five patients met inclusion criteria. Of these, 48 received G-CSF during all chemotherapy cycles for primary prophylaxis while 37 received G-CSF only for secondary prophylaxis. Same-day pegfilgrastim compared to next-day pegfilgrastim or filgrastim was the most common G-CSF dosing utilized with primary and secondary prophylaxis patients receiving it (87.5%, 94.6%) respectively. As shown in Table, primary and secondary prophylaxis groups were similar on baseline characteristics (p>0.05); the primary outcome of FN (p>0.05); all secondary outcomes (p>0.05) except for a higher frequency of dose delays in secondary (37.8%) vs primary prophylaxis patients (14.6%; p=0.01); and mean absolute neutrophil counts (ANC) in all cycles (p>0.05) except for cycles 3 and 5. Higher ANC levels were found in primary prophylaxis patients (4.06+0.43) vs secondary prophylaxis (3.03+0.30; p=0.03) for cycle 3 and (3.57+0.25) vs (2.88+0.26; p=0.03) for cycle 5.

Conclusion:

In this single-center retrospective study, BR-treated lymphoma and CLL patients receiving primary vs secondary with G-CSF showed similar outcomes except, notably, for chemotherapy dose delays that may put secondary patients at risk for poor treatment outcomes. Further research is needed to evaluate the impact of primary vs secondary prophylaxis on chemotherapy treatment outcomes.

Table

Disclosures

Persky: Sandoz: Consultancy; Debiopharm: Other: Member, Independent Data Monitoring Committee; Bayer: Consultancy; Morphosys: Other: Member, Independent Data Monitoring Committee. McBride:Sanofi Genzyme: Consultancy; Sandoz: Consultancy; teva: Consultancy.
Using Childhood International Prognostic Scoring in Pediatric Hodgkin Lymphoma Patients Treated with ABVD Can Mitigate Use of Interim PET in LMIC
Authors: Khedr Reham , Rashad Hanaa , Hamoda Asmaa Mohammed , Khorshed Eman , Elwakeel Madeha , Saad Mohamed , Mohamed Engy , Moussa Emad
Published: November 2019

Background & aim:Although 'interim' positron emission tomography (PET) may inform therapeutic decisions, Risk stratification at diagnosis allows earlier and potentially better modification during treatment of HL. the prognosis of HL in our center.In this study, we aimed to identify the prognostic role of both the IPET and validate Childhood Hodgkin International Prognostic Score (CHIPS) in predicting the prognosis of HL at our hospital.

Patients and methods:This is a retrospective, single-center study where a total of 1140 patients with newly diagnosed Hodgkin lymphoma were enrolled. Only 737 patients were eligible for analysis of both IPET and CHIPS scoring. PET scan was performed at baseline and after two cycles of chemotherapy (ABVD). Treatment was not changed according to the results of the interim scan. Stage 4 disease, large mediastinal mass, albumin (<3.5), and fever were each assigned one point in the CHIPS. Log-rank testing was used to compare EFS for each CHIPS (0-3) secondary analysis to detect EFS in relation to IPET was done.

Results : Only 737 patients were eligible for analysis of both IPET and CHIPS scoring. The 5-year EFS was 95% for patients with CHIPS=0, 94% for patients with CHIPS=1, 84.3% for patients with CHIPS=2, 82.4% for patients with CHIPS=3 and 70% for CHIPS=4 (P-value 0.001).The 5-year OS was 97.4% for patients with CHIPS=0, 97.9% for patients with CHIPS=1, 93.1% for patients with CHIPS=2, 94.7% for patients with CHIPS=3 and 85.6% for CHIPS=4 (P-value 0.039).

Slow early responders as detected by PET have 5-years EFS 73.8%, in comparison, those with evidence of response by PET had 5-year EFS of 91.3 %. Among this better risk patients, CHIPS can further classify patients; EFS 96.4% of patients with CHIPS 0 or 1 versus 85.8% for those with CHIPS 2, 3 or 4 who had an adverse outcome, even in the setting of response by PET (P<0.001).

Conclusion:

CHIPS is a good tool for predicting risk in patients with HL that may help to tailor therapy to risk factors known early at diagnosis. The benefit of using CHIPS versus early response PET scan suggests the feasibility of eliminating the costly PET scans, especially in LMIC.

Figure

Disclosures

No relevant conflicts of interest to declare.
Application of Single Cell Technologies to the Study of Hematopoiesis
Authors: Gottgens Bertie
Published: November 2019

The Gottgens group uses a combination of experimental and computational approaches to study how transcription factor networks control the function of blood stem cells and how mutations that perturb such networks cause diseases. The group's current research focuses on (i) single cell genomics of early blood development, (ii) computer models of the transcriptional landscape of blood stem cell differentiation, (iii) transcriptional consequences of leukaemogenic mutations, and (iv) molecular characterization of human blood stem cell populations used in cell and gene therapy protocols. As requested by the session chair, this year's presentation will first provide an overview of single cell technologies, and how they are advancing our understanding of multiple facets of haematology research. This will include single cell molecular profiling, as well as single cell functional assays, and in particular also how a combination of the two allows a more precise definition of haematopoietic stem and progenitor cell types. The rest of the presentation will focus on our multidisciplinary work combining single cell molecular profiling, bioinformatics analysis and experimental/functional validation to study the normal haematopoiesis, and contrast this with 6 mouse models of pre-leukaemic disease. Comprehensive bioinformatics analysis reveals not only qualitative changes in cellular abundance, but also pinpoints the underlying molecular changes that are most likely driving the early stages of malignant disease. An overarching theme will be how single cell landscapes allow us to move seamlessly between different scales of biological investigation, from the molecular to the cellular and whole tissue scale. Finally, extrapolation to human patient data demonstrates disease relevance of gene sets identified from comparative analysis of single cell transcriptional landscapes in mouse models.

Disclosures

Gottgens: Astra Zeneca: Research Funding; GSK: Research Funding; Novo Nordisk: Consultancy, Research Funding; Autolus: Consultancy, Research Funding.
Current perspectives on the role of interleukin-1 signalling in the pathogenesis of asthma and COPD
Authors: Osei Emmanuel T. , Brandsma Corry-Anke , Timens Wim , Heijink Irene H. , Hackett Tillie-Louise
Published: November 2019

Asthma and chronic obstructive pulmonary disease (COPD) cause significant morbidity and mortality worldwide. In the context of disease pathogenesis, both asthma and COPD involve chronic inflammation of the lung and are characterised by the abnormal release of inflammatory cytokines, dysregulated immune cell activity and remodelling of the airways. To date, current treatments still only manage symptoms and do not reverse the primary disease processes. In recent work, interleukin (IL)-1α and IL-1β have been suggested to play important roles in both asthma and COPD. In this review, we summarise overwhelming pre-clinical evidence for dysregulated signalling of IL-1α and IL-1β contributing to disease pathogenesis and discuss the paradox of IL-1 therapeutic studies in asthma and COPD. This is particularly important given recent completed and ongoing clinical trials with IL-1 biologics that have had varying degrees of failure and success as therapeutics for disease modification in asthma and COPD.
Optimization of Ohmic Contacts to p-GaAs Nanowires
Authors: Rizzo Piton Marcelo , Hakkarainen Teemu , Hilska Joonas , Koivusalo Eero , Lupo Donald , Galeti Helder Vinicius Avanço , Galvão Gobato Yara , Guina Mircea
Published: November 2019

The performance of Ohmic contacts applied to semiconductor nanowires (NWs) is an important aspect for enabling their use in electronic or optoelectronic devices. Due to the small dimensions and specific surface orientation of NWs, the standard processing technology widely developed for planar heterostructures cannot be directly applied. Here, we report on the fabrication and optimization of Pt/Ti/Pt/Au Ohmic contacts for p-type GaAs nanowires grown by molecular beam epitaxy. The devices were characterized by current–voltage (IV) measurements. The linearity of the IV characteristics curves of individual nanowires was optimized by adjusting the layout of the contact metal layers, the surface treatment prior to metal evaporation, and post-processing thermal annealing. Our results reveal that the contact resistance is remarkably decreased when a Pt layer is deposited on the GaAs nanowire prior to the traditional Ti/Pt/Au multilayer layout used for p-type planar GaAs. These findings are explained by an improved quality of the metal-GaAs interface, which was evidenced by grazing incidence X-ray diffraction measurements in similar metallic thin films deposited on GaAs (110) substrates. In particular, we show that Ti exhibits low degree of crystallinity when deposited on GaAs (110) surface which directly affects the contact resistance of the NW devices. The deposition of a thin Pt layer on the NWs prior to Ti/Pt/Au results in a 95% decrease in the total electrical resistance of Be-doped GaAs NWs which is associated to the higher degree of crystallinity of Pt than Ti when deposited directly on GaAs (110).
Proteomic and transcriptomic analyses of early and late-chronic Toxoplasma gondii infection shows novel and stage specific transcripts
Authors: Garfoot Andrew L. , Wilson Gary M. , Coon Joshua J. , Knoll Laura J.
Published: November 2019

Background

The protozoan pathogen Toxoplasma gondii has the unique ability to develop a chronic infection in the brain of its host by transitioning from the fast growing tachyzoite morphology to latent bradyzoite morphology. A hallmark of the bradyzoite is the development of neuronal cysts that are resilient against host immune response and current therapeutics. The bradyzoite parasites within the cyst have a carbohydrate and protein-rich wall and a slow-replication cycle, allowing them to remain hidden from the host. The intracellular, encysted lifestyle of T. gondii has made them recalcitrant to molecular analysis in vivo.

Results

Here, we detail the results from transcriptional and proteomic analyses of bradyzoite-enriched fractions isolated from mouse brains infected with T. gondii over a time course of 21 to 150 days. The enrichment procedure afforded consistent identification of over 2000 parasitic peptides from the mixed-organism sample, representing 366 T. gondii proteins at 28, 90, and 120 day timepoints. Deep sequencing of transcripts expressed during these three timepoints revealed that a subpopulation of genes that are transcriptionally expressed at a high level. Approximately one-third of these transcripts are more enriched during bradyzoite conditions compared to tachyzoites and approximately half are expressed at similar levels during each phase. The T. gondii transcript which increased the most over the course of chronic infection, sporoAMA1, shows stage specific isoform expression of the gene.

Conclusions

We have expanded the transcriptional profile of in vivo bradyzoites to 120 days post-infection and provided the first in vivo proteomic profile of T. gondii bradyzoites. The RNA sequencing depth of in vivo bradyzoite T. gondii was over 250-fold greater than previous reports and allowed us to identify low level transcripts and a novel bradyzoite-specific isoform of sporoAMA1.
Transcriptome analysis of Aspergillus niger xlnR and xkiA mutants grown on corn Stover and soybean hulls reveals a highly complex regulatory network
Authors: Khosravi Claire , Kowalczyk Joanna E. , Chroumpi Tania , Battaglia Evy , Aguilar Pontes Maria-Victoria , Peng Mao , Wiebenga Ad , Ng Vivian , Lipzen Anna , He Guifen , Bauer Diane , Grigoriev Igor V. , de Vries Ronald P.
Published: November 2019

Background

Enzymatic plant biomass degradation by fungi is a highly complex process and one of the leading challenges in developing a biobased economy. Some industrial fungi (e.g. Aspergillus niger) have a long history of use with respect to plant biomass degradation and for that reason have become ‘model’ species for this topic. A. niger is a major industrial enzyme producer that has a broad ability to degrade plant based polysaccharides. A. niger wild-type, the (hemi-)cellulolytic regulator (xlnR) and xylulokinase (xkiA1) mutant strains were grown on a monocot (corn stover, CS) and dicot (soybean hulls, SBH) substrate. The xkiA1 mutant is unable to utilize the pentoses D-xylose and L-arabinose and the polysaccharide xylan, and was previously shown to accumulate inducers for the (hemi-)cellulolytic transcriptional activator XlnR and the arabinanolytic transcriptional activator AraR in the presence of pentoses, resulting in overexpression of their target genes. The xlnR mutant has reduced growth on xylan and down-regulation of its target genes. The mutants therefore have a similar phenotype on xylan, but an opposite transcriptional effect. D-xylose and L-arabinose are the most abundant monosaccharides after D-glucose in nearly all plant-derived biomass materials. In this study we evaluated the effect of the xlnR and xkiA1 mutation during growth on two pentose-rich substrates by transcriptome analysis.

Results

Particular attention was given to CAZymes, metabolic pathways and transcription factors related to the plant biomass degradation. Genes coding for the main enzymes involved in plant biomass degradation were down-regulated at the beginning of the growth on CS and SBH. However, at a later time point, significant differences were found in the expression profiles of both mutants on CS compared to SBH.

Conclusion

This study demonstrates the high complexity of the plant biomass degradation process by fungi, by showing that mutant strains with fairly straightforward phenotypes on pure mono- and polysaccharides, have much less clear-cut phenotypes and transcriptomes on crude plant biomass.
Design of an Underwater Treadmill System for rehabilitation of older obese adults: a pre-post study
Authors: Kittichaikarn C. , Kuptniratsaikul V.
Published: November 2019

Background

Patients with knee osteoarthritis (OA) who are obese have problems performing land-based exercises. The reduced joint stress associated with aquatic exercise may benefit these patients. This study aimed to develop an underwater treadmill (UTM) machine that is affordable and suitable for use in developing countries, and to evaluate its efficacy in decreasing pain and increasing functional improvement.

Methods

Clinical testing of the UTM machine was performed in an outpatient setting at Siriraj Hospital during January–June 2017. Patients with knee OA, aged 50–85 years, numerical rating scale (NRS) ≥5/10, and body mass index (BMI) ≥25 kg/m 2 were recruited. The UTM exercise protocol was 30 min/session, 3 days/week, for 4 weeks. The main outcomes were NRS pain score, 6-min walk distance (6MWD), quadriceps strength (QS) and body weight. Those outcomes were evaluated at baseline and at week 4.

Results

The UTM was constructed with safety, ergonomically designed and user-friendly control panel with push button for emergency stopping. Thirty patients were included for clinical testing. The mean age was 62.8 years, and almost all were female. The mean BMI was 28.9 kg/m 2. Most patients (65.0%) developed bilateral knee OA, used pain medication (56.7%), and engaged in regular knee exercise (73.3%). Of the 30 enrolled patients, 6 withdrew. All of the remaining 24 patients attended all 12 sessions. The mean difference between baseline and the end of the study was − 2.3 (95% CI: − 3.0, − 1.5; p < 0.001) for NRS pain; 34.9 m (95% CI: 14.1, 55.8; p = 0.002) for 6MWD; and, 1.8 kg (95% CI, 1.1, 2.6; p < 0.001) for QS. Concerning adverse events, 4 patients (15.4%) developed muscle pain, 2 patients (7.7%) had joint pain, and 1 patient (3.9%) withdrew due to severe knee pain. Two-thirds of patients described themselves as being ‘very satisfied’ with UTM exercise, and approximately 90% of participants rated their symptoms as ‘improved’ or ‘much improved’.

Conclusions

4-week exercise with UTM can significantly improve NRS pain, 6MWD, and QS. UTM could be an alternative treatment for patients with knee OA who are obese due to small size, durability, and ecofriendly design as an exercise modality.
Comparative efficacy of pharmacological and nonpharmacological treatments for chronic idiopathic constipation in China: a Bayesian network meta-analysis
Authors: Shi Qingyang , Tan Lizi , Liu Chunxiang , Wang Huijun , Zhang Junhua , Wang Hui , Zhai Jingbo
Published: November 2019

Background

To provide evidence for medical management of chronic idiopathic constipation (CIC) in China based on comparisons of all clinical practical interventions using Bayesian network meta-analysis.

Methods

We conducted a systematic literature review by searching PubMed, Embase, Cochrane Central, the China National Knowledge Infrastructure (CNKI), and the Wanfang Database (inception to May 2019) for randomized controlled trials (RCTs) for CIC in Chinese people. Only RCTs that recruited participants aged over 18 and diagnosed with CIC by the Rome II, III or IV criteria were included. We used three outcomes to examine efficacy. The risk ratio (RR) of the responder rate, based on ≥3 spontaneous bowel movements (SBMs) per week after treatment, was the primary outcome, and the SBM count per week and the Bristol score (BS) were secondary outcomes. In addition, adverse effects (AEs) were also considered a secondary outcome to evaluate safety. We conducted Bayesian network meta-analysis with random effects, and the RR or mean difference with its 95% credible interval was calculated. In addition, we ranked all treatments via their cumulative curves (SUCRA) and assessed the quality of evidence according to the GRADE criteria.

Results

We included a total of 42 trials (6820 participants) of 20 grouped interventions that included pharmacological and nonpharmacological treatments. For the primary outcome, fourteen interventions were significantly better than placebo, and Probiotics plus Mosapride (PB + MP) appeared superior to others (GRADE quality of evidence: Moderate to Low), followed by Prucalopride (PP) (High to Low) and Electroacupuncture (EA) (High to Low). For SBM, Compound sodium bicarbonate suppository (CSBS) appeared to be best, with an SUCRA value of 90% (High to Low). For BS, Lactulose plus Probiotics (LT + PB) was superior to others (Moderate to Low), followed by Polyethylene glycol (PEG) (High to Moderate). Although all interventions appeared non-significant when compared with placebo in terms of adverse effects, Lactulose plus Mosapride showed greater risk than others on ranking probability.

Conclusions

Given the GRADE assessment, PB + MP, PP and EA may be the priory options with moderate certainty in the quality of evidence for the primary outcome. For SBM, a CSBS may be the best option with moderate certainty in the quality of evidence. For BS, PEG may be the priory option with high certainty in the quality of evidence. However, due to a lack of high certainty in the quality of evidence, caution is needed when recommending the interventions. Because of the limitations, an increased number of trials are required for more accurate results.
Knowledge, attitude, and perceived barriers regarding colorectal cancer screening practices and risk factors among medical students in Saudi Arabia
Authors: Althobaiti Asma , Jradi Hoda
Published: November 2019

Background

Colorectal cancer (CRC) is a major health problem. It is the third most diagnosed common tumour and the fourth leading cause of cancer-related deaths worldwide. Early screening has been shown to decrease the incidence of CRC cancer and decrease mortality. In Saudi Arabia (SA), there is no national policy for CRC screening despite the growing incidence of the disease. This study investigated the knowledge of risk factors for CRC, recommendations for screening, and attitudes and barriers towards screening among medical students.

Methods

Data was collected using a self-administered valid and reliable questionnaire consisting of demographic characteristics, knowledge, attitude, and barriers measurements completed by 581 medical students from two Saudi schools. Frequencies and mean scores of knowledge and attitude were determined. The likelihood of students having adequate knowledge of CRC risk factors and screening modalities was estimated using multivariate logistic regression analysis.

Results

Knowledge of the risk factors for CRC and screening modalities, and attitude towards screening were poor in 52.47 and 57.83% of the surveyed medical students; respectively. Higher level of medical education (OR = 3.23; 95% CI: 2.01–5.18) and a positive attitude towards CRC screening (OR = 2.74; 95% CI: 1.86–4.03) were independent predictors of higher knowledge levels. Lack of awareness about CRC and screening modalities among patients, and shortage of specialized healthcare providers were barriers independently associated with low knowledge levels.

Conclusions

Saudi medical students have limited knowledge of CRC risk factors of and a poor attitude towards CRC screening. These results contribute to our understanding of missed teaching opportunities in Saudi medical schools and suggest intervening at the medical school, clinical practice, and population levels to increase CRC screening practices.
Identification of novel cell glycolysis related gene signature predicting survival in patients with endometrial cancer
Authors: Wang Zi-Hao , Zhang Yun-Zheng , Wang Yu-Shan , Ma Xiao-Xin
Published: November 2019

Background

Endometrial cancer (EC) is one of the three major gynecological malignancies. Numerous biomarkers that may be associated with survival and prognosis have been identified through database mining in previous studies. However, the predictive ability of single-gene biomarkers is not sufficiently specific. Genetic signatures may be an improved option for prediction. This study aimed to explore data from The Cancer Genome Atlas (TCGA) to identify a new genetic signature for predicting the prognosis of EC.

Methods

mRNA expression profiling was performed in a group of patients with EC (n = 548) from TCGA. Gene set enrichment analysis was performed to identify gene sets that were significantly different between EC tissues and normal tissues. Cox proportional hazards regression models were used to identify genes significantly associated with overall survival. Quantitative real-time-PCR was used to verify the reliability of the expression of selected mRNAs. Subsequent multivariate Cox regression analysis was used to establish a prognostic risk parameter formula. Kaplan–Meier survival estimates and the log‐rank test were used to validate the significance of risk parameters for prognosis prediction.

Result

Nine genes associated with glycolysis (CLDN9, B4GALT1, GMPPB, B4GALT4, AK4, CHST6, PC, GPC1, and SRD5A3) were found to be significantly related to overall survival. The results of mRNA expression analysis by PCR were consistent with those of bioinformatics analysis. Based on the nine-gene signature, the 548 patients with EC were divided into high/low-risk subgroups. The prognostic ability of the nine-gene signature was not affected by other factors.

Conclusion

A nine-gene signature associated with cellular glycolysis for predicting the survival of patients with EC was developed. The findings provide insight into the mechanisms of cellular glycolysis and identification of patients with poor prognosis in EC.
Mesangial cells are key contributors to the fibrotic damage seen in the lupus nephritis glomerulus
Authors: Wright Rachael D. , Dimou Paraskevi , Northey Sarah J. , Beresford Michael W.
Published: November 2019

Background

Lupus nephritis (LN) affects up to 80% of juvenile-onset systemic lupus erythematosus patients. Mesangial cells (MCs) comprise a third of the glomerular cells and are key contributors to fibrotic changes within the kidney. This project aims to identify the roles of MCs in an in vitro model of LN.

Methods

Conditionally immortalised MCs were treated with pro-inflammatory cytokines or with patient sera in an in vitro model of LN and assessed for their roles in inflammation and fibrosis.

Results

MCs were shown to produce pro-inflammatory cytokines in response to a model of the inflammatory environment in LN. Further the cells expressed increased levels of mRNA for extracellular matrix (ECM) proteins (COL1A1, COL1A2, COL4A1andLAMB1), matrix metalloproteinase enzymes (MMP9) and tissue inhibitors of matrix metalloproteinases (TIMP1). Treatment of MCs with serum from patients with active LN was able to induce a similar, albeit milder phenotype. Treatment of MCs with cytokines or patient sera was able to induce secretion of TGF-β1, a known inducer of fibrotic changes. Inhibition of TGF-β1 actions through SB-431542 (an activin A receptor type II-like kinase (ALK5) inhibitor) was able to reduce these responses suggesting that the release of TGF-β1 plays a role in these changes.

Conclusions

MCs contribute to the inflammatory environment in LN by producing cytokines involved in leukocyte recruitment, activation and maturation. Further the cells remodel the ECM via protein deposition and enzymatic degradation. This occurs through the actions of TGF-β1 on its receptor, ALK5. This may represent a potential therapeutic target for treatment of LN-associated fibrosis.