bluuebunny/crossref_metadata_2025 huggingface dataset:
The H1047R PIK3CA oncogene induces a senescence-like state, pleiotropy and acute HSP90 dependency in HER2+ mammary epithelial cells
Authors: Chakrabarty Anindita , Surendran Sreeraj , Bhola Neil E , Mishra Vishnu S , Wani Tasaduq Hussain , Baghel Khemraj S , Arteaga Carlos L , Garg Rohini , Chowdhury Goutam
Published: June 2019

In pre-clinical models, co-existence of Human Epidermal Growth Factor Receptor-2 (HER2)-amplification and PI3K catalytic subunit (PIK3CA) mutations results in aggressive, anti-HER2 therapy-resistant breast tumors. This is not always reflected in clinical setting. We speculated that the complex interaction between the HER2 and PIK3CA oncogenes is responsible for such inconsistency. We performed series of biochemical, molecular and cellular assays on genetically engineered isogenic mammary epithelial cell lines and breast cancer cells expressing both oncogenes. In vitro observations were validated in xenografts models. We showed that H1047R, one of the most common PIK3CA mutations, is responsible for endowing a senescence-like state in mammary epithelial cells overexpressing HER2. Instead of imposing a permanent growth arrest characteristic of oncogene-induced senescence, the proteome secreted by the mutant cells promotes stem cell enrichment, angiogenesis, epithelial-to-mesenchymal transition, altered immune surveillance and acute vulnerability toward HSP90 inhibition. We inferred that the pleiotropism, as observed here, conferred by the mutated oncogene, depending on the host microenvironment, contributes to conflicting pre-clinical and clinical characteristics of HER2+, mutated PIK3CA-bearing tumor cells. We also came up with a plausible model for evolution of breast tumors from mammary epithelial cells harboring these two molecular lesions.
Performance of soluble Klotho assays in clinical samples of kidney disease
Authors: Neyra Javier A , Moe Orson W , Pastor Johanne , Gianella Fabiola , Sidhu Sachdev S , Sarnak Mark J , Ix Joachim H , Drew David A
Published: June 2019

Background

Soluble Klotho has multiple systemic salutary effects. In animals, both acute and chronic kidney disease models display systemic Klotho deficiency. As such, there is considerable interest in investigating soluble Klotho as a biomarker in patients with different types and severity of kidney diseases. Unfortunately, there remains uncertainty regarding the best method to measure soluble Klotho in human serum samples.

Methods

Using human serum samples obtained from several clinical cohorts with a wide range of kidney function, we measured soluble Klotho using a commercial enzyme-linked immunosorbent assay (ELISA) as well as with an immunoprecipitation–immunoblot (IP–IB) assay utilizing a synthetic antibody with high affinity and specificity for Klotho. Recovery of spiking with a known amount of exogenous Klotho was tested. A subset of samples was analyzed with and without the addition of a protease inhibitor cocktail at the time of collection or after the first freeze–thaw cycle to determine if these maneuvers influenced performance.

Results

The IP–IB assay was superior to the ELISA at recovery of exogenous Klotho (81–115% versus 60–81%) across the spectrum of kidney function. Klotho measurements by IP–IB were highly correlated with estimated glomerular filtration rate (eGFR) (R = 0.80, P < 0.001) in comparison with the commercial ELISA, which exhibited minimal correlation with eGFR (R = 0.18, P = 0.12). Use of a protease inhibitor cocktail neither improved nor impaired performance of the IP–IB assay; however, subsequent freeze–thaw cycle resulted in a significant reduction in Klotho recovery and dissipated the correlation between Klotho levels and eGFR. With the ELISA, the use of protease inhibitor cocktail resulted in an increase in intrasubject variability.

Conclusions

The IP–IB assay is preferable to the commercial ELISA to measure soluble Klotho concentrations in never-thawed serum samples of humans with varying severity of kidney disease. However, due to the labor-intensive nature of the IP–IB assay, further research is needed to secure an assay suitable for high-throughput work.
Calciprotein particles and fibroblast growth factor 23 contribute to the pathophysiology of hypercalcemia in a patient with renal sarcoidosis
Authors: Iwazu Yoshitaka , Kuro-o Makoto , Miura Yutaka , Takeda Shin-ichi , Yamada Toshiyuki , Nagata Daisuke
Published: June 2019

In patients with sarcoidosis, dysregulated calcium metabolism is one of the frequently observed complications. However, little attention has been paid to abnormal phosphate metabolism. Herein we present the case of a 42-year-old Japanese man with renal sarcoidosis who developed acute kidney injury due to hypercalcemia and nephrolithiasis. Laboratory data showed hypercalcemia with a normal serum phosphate level and high serum 1,25-hydroxyvitamin D3, fibroblast growth factor 23 (FGF23) and calciprotein particle (CPP) levels. After treatment with oral prednisone and bisphosphonate, the laboratory abnormalities and renal dysfunction were resolved. Thus increases in FGF23 and CPP may indicate disturbed phosphate metabolism in renal sarcoidosis.
A Markov chain analysis of the dynamics of homophily
Authors: Guerrero Katerine , Finke Jorge
Published: June 2019

Many networks are made up of different nodes types, which are determined by a set of common quantitative or qualitative node properties. Understanding the effects of homophilic relationships, that is, the tendency of nodes to establish links to other nodes that are alike, requires formal frameworks that explain how local decision-making mechanisms contribute to the formation of particular network structures. Based on two simple stochastic mechanisms for establishing links, this article introduces a model that explains the emergence of homophily as an aggregate group and network level outcome. We characterize the dynamics of homophily and present conditions that guarantee that the amount of homophily exceeds the expected amount of a purely random decision-making process. Moreover, we show that the proposed model resembles patterns of homophily in a citation network of political blogs. Finally, we use the model to design a non-homophilic node detection algorithm for identifying nodes that establish connections without a particular preference for either node type.
C-type lectin receptor-mediated immune recognition and response of the microbiota in the gut
Authors: Li Tian-Hang , Liu Ling , Hou Ya-Yi , Shen Su-Nan , Wang Ting-Ting
Published: June 2019

C-type lectin receptors (CLRs) are powerful pattern-recognition receptors that discern ‘self’ and ‘non-self’ in our body and protect us from invasive pathogens by mediating immune recognition and response. The gastrointestinal tract is very important for the maintenance of homeostasis; it is the largest shelter for the billions of microorganisms in the body and CLRs play a crucial regulatory role in this system. This study focuses on several CLRs, including Dectin-1, Dectin-2, Dectin-3 and Mincle. We summarize the roles of CLRs in maintaining gastrointestinal immune-system homeostasis, especially their functions in mediating immune recognition and responses in the gut, discuss their relationships to some diseases, highlight the significance of CLR-mediated sensing of microbial and non-microbial compounds in the gut immune system and identify new therapeutic targets.
First Estimation of the Spontaneous Mutation Rate in Diatoms
Authors: Krasovec Marc , Sanchez-Brosseau Sophie , Piganeau Gwenael
Published: June 2019

Mutations are the origin of genetic diversity, and the mutation rate is a fundamental parameter to understand all aspects of molecular evolution. The combination of mutation–accumulation experiments and high-throughput sequencing enabled the estimation of mutation rates in most model organisms, but several major eukaryotic lineages remain unexplored. Here, we report the first estimation of the spontaneous mutation rate in a model unicellular eukaryote from the Stramenopile kingdom, the diatom Phaeodactylum tricornutum (strain RCC2967). We sequenced 36 mutation accumulation lines for an average of 181 generations per line and identified 156 de novo mutations. The base substitution mutation rate per site per generation is μbs = 4.77 × 10−10 and the insertion–deletion mutation rate is μid = 1.58 × 10−11. The mutation rate varies as a function of the nucleotide context and is biased toward an excess of mutations from GC to AT, consistent with previous observations in other species. Interestingly, the mutation rates between the genomes of organelles and the nucleus differ, with a significantly higher mutation rate in the mitochondria. This confirms previous claims based on indirect estimations of the mutation rate in mitochondria of photosynthetic eukaryotes that acquired their plastid through a secondary endosymbiosis. This novel estimate enables us to infer the effective population size of P. tricornutum to be Ne∼8.72 × 106.
Quantifying the estimation error of principal component vectors
Authors: Hauser Raphael , Lember Jüri , Matzinger Heinrich , Kangro Raul
Published: June 2019

Principal component analysis (PCA) is an important pattern recognition and dimensionality reduction tool in many applications. Principal components are computed as eigenvectors of a maximum likelihood covariance $\widehat{\varSigma }$ that approximates a population covariance $\varSigma$, and these eigenvectors are often used to extract structural information about the variables (or attributes) of the studied population. Since PCA is based on the eigendecomposition of the proxy covariance $\widehat{\varSigma }$ rather than the ground-truth $\varSigma$, it is important to understand the approximation error in each individual eigenvector as a function of the number of available samples. The combination of recent results of Koltchinskii & Lounici (2017, Bernoulli, 23, 110–133) and Yu et al. (2015, Biometrika, 102, 315–323) yields such bounds. In the present paper we sharpen these bounds and show that eigenvectors can often be reconstructed to a required accuracy from a sample of strictly smaller size order.
First Record of Quill Mites (Acariformes: Syringophilidae: Picobiinae) Living in the Quill Walls of Parrots
Authors: Skoracki Maciej , Sikora Bozena , Hromada Martin
Published: June 2019

Quill mites (Acariformes: Prostigmata: Syringophilidae) are a family of about 400 ectoparasite species known to infest birds for over 140 yr. All previously identified quill mites occur in the calamus, a cavity of the bird feather, where they spent their entire life except for a short dispersion period. In this study, we report the first record of quill mites dwelling in the quill walls during their entire life cycle. In short, we found all of the different life stages of the species Lawrencipicobia poicephali (Skoracki & Dabert 2002) (Acariformes: Syringophilidae) in the quill wall in 33 host individuals of six different host species from many different locations in Sub-Saharan Africa. Hosts were collected over a time period spanning more than 70 yr. Lastly, we report six new avian hosts for this ectoparasite species.
Dark Hope: Notes on Uncertain Futures
Authors: Van Horn O’neil
Published: June 2019

In the midst of the Anthropocene, what might it mean to have hope? This article intends not to ‘answer’ such a question, but instead sketch possibilities for an alternative, cloudier, indeed livable understanding of ‘hope’. As an exercise in theopoetics, this piece offers the concept of ‘dark hope’ as a viable symbol for the purpose of eco-activisms—entangled, intersectional. To achieve a blurred illustration of this vision of ‘dark hope’, this article will trace relations between Karmen MacKendrick’s work on the haunting affects of memory, Paul Tillich’s confrontation with ‘non-being’, and queer visions of possible, and ever troubling, futures on a rapidly shifting planet. If nothing else, this project strives toward an understanding of hope that can stand up to the woes of anthropogenic climate change, that can be practiced in the face of disaster, that can be lived even in the midst of ruins.
The evolution of sizes and specific angular momenta in hierarchical models of galaxy formation and evolution
Authors: Zoldan Anna , De Lucia Gabriella , Xie Lizhi , Fontanot Fabio , Hirschmann Michaela
Published: June 2019

We extend our previous work focused at z ∼ 0, studying the redshift evolution of galaxy dynamical properties using the state-of-the-art semi-analytic model GAEA (GAlaxy Evolution and Assembly): we show that the predicted size–mass relation for discy/star-forming and quiescent galaxies is in good agreement with observational estimates, up to z ∼ 2. Bulge-dominated galaxies have sizes that are offset low with respect to observational estimates, mainly due to our implementation of disc instability at high redshift. At large masses, both quiescent and bulge-dominated galaxies have sizes smaller than observed. We interpret this as a consequence of our most massive galaxies having larger gas masses than observed, and therefore being more affected by dissipation. We argue that a proper treatment of quasar-driven winds is needed to alleviate this problem. Our model compact galaxies have number densities in agreement with observational estimates and they form most of their stars in small and low angular momentum high-z haloes. GAEA predicts that a significant fraction of compact galaxies forming at high-z is bound to merge with larger structures at lower redshifts: therefore they are not the progenitors of normal-size passive galaxies at z = 0. Our model also predicts a stellar–halo size relation that is in good agreement with observational estimates. The ratio between stellar size and halo size is proportional to the halo spin and does not depend on stellar mass but for the most massive galaxies, where active galactic nucleus feedback leads to a significant decrease of the retention factor (from about 80 per cent to 20 per cent).
Interplay of transcriptional signaling by progesterone, cyclic AMP, and inflammation in myometrial cells: implications for the control of human parturition
Authors: Stanfield Zachary , Amini Peyvand , Wang Junye , Yi Lijuan , Tan Huiqing , Chance Mark R , Koyutürk Mehmet , Mesiano Sam
Published: June 2019

Parturition involves cellular signaling changes driven by the complex interplay between progesterone (P4), inflammation, and the cyclic adenosine monophosphate (cAMP) pathway. To characterize this interplay, we performed comprehensive transcriptomic studies utilizing eight treatment combinations on myometrial cell lines and tissue samples from pregnant women. We performed genome-wide RNA-sequencing on the hTERT-HM${}^{A/B}$ cell line treated with all combinations of P4, forskolin (FSK) (induces cAMP), and interleukin-1$\beta$ (IL-1$\beta$). We then performed gene set enrichment and regulatory network analyses to identify pathways commonly, differentially, or synergistically regulated by these treatments. Finally, we used tissue similarity index (TSI) to characterize the correspondence between cell lines and tissue phenotypes. We observed that in addition to their individual anti-inflammatory effects, P4 and cAMP synergistically blocked specific inflammatory pathways/regulators including STAT3/6, CEBPA/B, and OCT1/7, but not NF$\kappa$B. TSI analysis indicated that FSK + P4- and IL-1$\beta$-treated cells exhibit transcriptional signatures highly similar to non-laboring and laboring term myometrium, respectively. Our results identify potential therapeutic targets to prevent preterm birth and show that the hTERT-HM${}^{A/B}$ cell line provides an accurate transcriptional model for term myometrial tissue.
CNBP controls transcription by unfolding DNA G-quadruplex structures
Authors: David Aldana P , Pipier Angélique , Pascutti Federico , Binolfi Andrés , Weiner Andrea M J , Challier Emilse , Heckel Sofía , Calsou Patrick , Gomez Dennis , Calcaterra Nora B , Armas Pablo
Published: June 2019

Guanine-rich DNA strands can fold into non-canonical four-stranded secondary structures named G-quadruplexes (G4). Experimental evidences suggest that G4-DNA surrounding transcription start sites act as cis-regulatory elements by either stimulating or inhibiting gene transcription. Therefore, proteins able to target and regulate specific G4 formation/unfolding are crucial for G4-mediated transcriptional control. Here we present data revealing that CNBP acts in vitro as a G4-unfolding protein over a tetramolecular G4 formed by the TG4T oligonucleotide, as well as over the G4 folded in the promoters of several oncogenes. CNBP depletion in cellulo led to a reduction in the transcription of endogenous KRAS, suggesting a regulatory role of CNBP in relieving the transcriptional abrogation due to G4 formation. CNBP activity was also assayed over the evolutionary conserved G4 enhancing the transcription of NOGGIN (NOG) developmental gene. CNBP unfolded in vitro NOG G4 and experiments performed in cellulo and in vivo in developing zebrafish showed a repressive role of CNBP on the transcription of this gene by G4 unwinding. Our results shed light on the mechanisms underlying CNBP way of action, as well as reinforce the notion about the existence and function of G4s in whole living organisms.
Trends in Diagnosed Chronic Hepatitis B in a US Health System Population, 2006–2015
Authors: Lu Mei , Zhou Yueren , Holmberg Scott D , Moorman Anne C , Spradling Philip R , Teshale Eyasu H , Boscarino Joseph A , Daida Yihe G , Schmidt Mark A , Li Jia , Rupp Loralee B , Trudeau Sheri , Gordon Stuart C ,   Chronic Hepatitis Cohort Study Investigators
Published: June 2019

Background

Trends in the epidemiology of chronic hepatitis B (CHB) among routine clinical care patients in the United States are not well documented. We used data from the Chronic Hepatitis Cohort Study to investigate changes in prevalence and newly recorded cases of CHB from 2006 to 2015.

Methods

Annual percentage changes (APCs) were estimated using join point Poisson regression. Analyses were adjusted by study site; when an interaction with the trend was observed, APCs were estimated by subgroups. Differences in rates based on race, age, and sex were calculated with rate ratios.

Results

We identified 5492 patients with CHB within select health systems with total populations that ranged from 1.9 to 2.4 million persons. From 2006 to 2014, the prevalence of diagnosed CHB increased from 181.3 to 253.0 per 100 000 persons in the health system population; from 2014 to 2015, it declined to 237.0 per 100 000 persons. APC was +3.7%/y through 131 December 2014 (P < .001) and −15.0%/y (P < .001) thereafter. The rate of newly reported cases of CHB did not change significantly across the study period (APC, −1.1%/y; P = .07). The rates of newly reported cases were 20.5 times higher among patients in the Asian American/American Indian/Pacific Islander (ASINPI) category, compared with white patients, and 2.8 times higher among African American patients. The ratio of male to female patients was roughly 3:2.

Conclusions

The prevalence of diagnosed CHB in this US patient population increased from 2006 to 2014, after which it decreased significantly. Rates declined most rapidly among patients ≤40 or 61–70 years old, as well as among ASINPI patients. The rate of newly reported cases remained steady over the study period.
Poisson vs. Gaussian statistics for sparse X-ray data: Application to the soft X-ray spectrometer
Authors: Yamada Shinya , Axelsson Magnus , Ishisaki Yoshitaka , Konami Saori , Takemura Nozomi , Kelley Richard L , Kilbourne Caroline A , Leutenegger Maurice A , Porter F Scott , Eckart Megan E , Szymkowiak Andrew
Published: June 2019

Reliable results when fitting X-ray data require proper consideration of the statistics involved. We probe the impact of Gaussian versus Poisson statistics at low count levels using both the standard χ2 method and maximum likelihood based on Poisson (C) statistics. The difference is studied and quantified through simulated spectra with known properties. We then test the results through analysis of Mn Kα calibration data taken with the flight spare microcalorimeter for the Hitomi soft X-ray spectrometer. Through comparison with simulations, our results show that the χ2 method tends to give overly optimistic estimates of the detector energy resolution, in particular when there are few counts. Given an energy resolution of ∼5 eV and a line with about 100 photons, the line width becomes ∼10% lower in the χ2 method than in Poisson statistics. This is a consequence of the uncertainties being dominated by counting statistics, and therefore highlights the need to choose the appropriate fit statistic.
A case report and literature review: Factitious disorder imposed on another and malingering by proxy
Authors: Walters Ilana C , MacIntosh Rachel , Blake Kim D
Published: June 2019

Factitious disorder imposed on another (FDIA) and malingering by proxy (MAL-BP) are two forms of underreported child maltreatment that should remain on physicians’ differential. This case of a 2-year-old boy, which spans 6 years, reveals the complexity in and difficulties with diagnosis. Key features include the patient’s mother using advanced medical jargon to report multiple disconnected concerns and visits to numerous providers. As a result, the patient underwent many investigations which often revealed normal findings. FDIA was suspected by the paediatrician, especially following corroboration with the child’s day care and past primary health care provider. This case demonstrates the possible overlap in diagnoses, which are characterized by a lack of consistent presentation and deceitful caregivers, often complicated by true underlying illness. The authors use clinical experience and limited existing literature to empower paediatricians to confidently diagnose and report FDIA and MAL-BP to limit future harm to children.
From Research Partner to Embedded Criminologist: A Synthesized Taxonomy and Reflections From the Field
Authors: Guerette Rob T , Lee-Silcox Joelle , Przeszlowski Kimberly
Published: June 2019

Perhaps never before has the need for collaboration between academic researchers and police agencies been more salient. Technological advances have brought advanced hardware systems into daily policing operations, and analytical software and information systems have assembled massive volumes of data repositories. To produce usable knowledge from these information troves, advanced analytical knowledge has become necessary. At the same time, the evidence-based policy movement requires that police agencies equip themselves with the ability to interpret and produce evaluations of new policing initiatives using sound scientific evaluation methods, something which also requires specialized and advanced knowledge not usually housed within policing organizations. In response to these demands, researcher–police collaborations have become more prevalent. Yet, the specific nature of these researcher–police relationships can take many forms. This article synthesizes previous literature and develops a taxonomy for classifying researcher–police partnerships and discusses the experiences of moving from an external research partner to an embedded criminological ‘unit’ within the Miami Police Department.
Deformation of neutron stars due to poloidal magnetic fields
Authors: Yanase K , Yoshinaga N , Nakano E , Watanabe C
Published: June 2019

The mass–radius (MR) relation of deformed neutron stars in the axially symmetric poloidal magnetic field is calculated. The MR relation is obtained by solving the Hartle equations, whereas the one for spherical stars is obtained by the Tolman–Oppenheimer–Volkoff equations. The anisotropic effects of the poloidal magnetic fields are found to be non-negligible for a strong magnetic field more than $3\times10^{18}$ G at the center of a neutron star.
Changes in Public Opinion Polling: How Newspaper Polls Gained Political Clout
Authors: MAEDA Yukio
Published: June 2019

Approval in public opinion polling is an important resource for prime ministers in contemporary Japanese politics. Though it is widely agreed that cabinet approval ratings increased in political significance after the electoral reform and the restructuring of government agencies, little attention has been paid to the fact that public opinion polling in the mass media also experienced a transformation during the same period. This article surveys the history of public opinion polling since 1945. It argues that changes in survey methodology enabled newspapers to conduct and report opinion polling quickly and frequently, which has strengthened the link between approval ratings and ongoing political processes.
Enhancing the adoption of stockpiling tall fescue and managed grazing
Authors: Freeman Sharon , Poore Matthew , Shaeffer April
Published: June 2019

One of the primary goals of extension is to encourage and support adoption of improved technologies. Managed grazing and stockpiling forage are two such technologies. The goal of this project was to encourage adoption of these practices by providing on-farm demonstrations of the technologies. We also collected forage and soil data and held workshops at each of the demonstration sites. Host producers were selected, given a basic kit of temporary fencing supplies for use during the demonstration, and instructed on their proper use during the winter stockpiling season. Forage yield and quality data were collected and soil tests made to show the economic advantages of proper fertilization and grazing fresh forage in contrast to feeding stored forage and concentrate. The nutritive value of the forage stockpiled in September through November (67% total digestible nutrients [TDN] and 14% crude protein, CP) exceeded the needs of the cattle and was greater than the nutrient content of hay present on the farms (59% TDN and 11% CP, P < 0.01). The mean quantity of available forage (2,856 ± 164 kg dry matter per hectare) provided an average of 260 (±81.8) standard cow (545 kg) grazing days per hectare of stockpiled forage. Taking into account the higher nutritive value of the fresh forage when compared with hay and the savings of time and equipment costs by grazing, we estimated that grazing stockpiled forage saved $1.28 per standard cow per day. The grazing management skills gained during this project and the temporary fencing technology were adopted by 93% of the demonstration farms that responded to our follow-up survey (78% of demonstration sites), and the area managed with these technologies increased on these sites more than 350%. Having the hosts share personal experiences played an important role in encouraging their peers to adopt the technologies.
Get Rich or Die Tryin’: Perceived Earnings, Perceived Mortality Rates, and Migration Decisions of Potential Work Migrants from Nepal
Authors: Shrestha Maheshwor
Published: June 2019

This article reports on a randomized field experiment in which potential work migrants from Nepal to Malaysia and the Persian Gulf countries are provided with information on wages and mortality incidences at their intended destinations. It is found that, particularly for the group of potential migrants without prior foreign migration experience, the information changes their expectations of earnings and mortality risks abroad, which further changes their actual migration decisions. Using the exogenous variation in expectations, it is estimated that the elasticity of migration with respect to mortality rate expectation is 0.8, and the elasticity of migration with respect to earnings expectation is 1.1.
S118 IMPACT OF PRE‐TRANSPLANT DONOR‐SPECIFIC ANTI‐HLA ANTIBODIES IN CORD BLOOD TRANSPLANTATION ON BEHALF OF THE TRANSPLANT COMPLICATIONS WORKING GROUP OF JSHCT
Authors: Fuji S. , Oshima K. , Ohashi K. , Sawa M. , Saito K. , Eto T. , Tanaka M. , Onizuka M. , Nakamae H. , Shiratori S. , Ozawa Y. , Hidaka M. , Nagamura‐Inoue T. , Tanaka H. , Fukuda T. , Ichinohe T. , Atsuta Y. , Ogata M.
Published: June 2019

Background:

Cord blood transplant (CBT) is an effective treatment for hematological diseases. Graft failure (GF) remains a major complication of CBT. Although presence of pre‐transplant donor‐specific anti‐HLA antibody (DSA) was reported to be associated with an increased risk of GF after CBT, data is still limited.

Aims:

In order to clarify the impact of pre‐transplant DSA, we conducted a retrospective analysis of recipients of CBT with pre‐transplant anti‐HLA antibody using the database of Japan Society for Hematopoietic Cell Transplantation (JSHCT).

Methods:

Data for recipients of CBT with pre‐transplant anti‐HLA antibody from 2010 to 2014 were obtained from the Transplant Registry Unified Management Program. Additional data such as detailed information about anti‐HLA antibody were collected. This study was approved by the institutional review boards of the JSHCT and Oita University.

Results:

In total, 343 patients who received CBT with detailed information about anti‐HLA antibody were included in the further analysis. Median age was 51 years (range, 0 to 71). Regarding DSA, 25 patients had mean fluorescence intensity (MFI) >1000 (DSA‐positive group) and 318 patients had MFI<1000 (DSA‐negative group). Cumulative incidence of neutrophil engraftment at 60 days after CBT was 75.7% (95% CI, 70.6–80.1) in the DSA‐negative group and 56.0% (95% CI, 34.1–73.1) in the DSA‐positive group (P = 0.03, Figure). Grouped according to the HLA class of DSA, cumulative incidence of neutrophil engraftment was 71.4% in patients with DSA against HLA class II antigen (n = 14), 44.4% in those with DSA against HLA class I antigen (n = 9), 0% in those with DSA against both HLA class I and class II antigens (n = 2). Stratified according to the achievement of neutrophil engraftment, the probabilities of overall survival at 1 year after CBT were 62.7% (95% CI, 56.2–68.5) in the engrafted patients and 19.5% (11.8–28.5) in the non‐engrafted patients (P < 0.001).

Summary/Conclusion:

Our study is a largest cohort of CBT recipients with pre‐transplant anti‐HLA antibody. Pre‐transplant DSA was associated with an increased risk of GF. The importance of target HLA antigen should be clarified in the future study.

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PF230 THE RNA HELICASE DDX17 PROTECTS ACUTE MYELOID LEUKEMIA CELLS AGAINST DNA DAMAGE
Authors: Jagdhane P. , Garg S. , Xia J. , He L. , Misiak D. , Mortusewicz O. , Helleday T. , Müller‐Tidow C. , Koehn M. , Pabst C.
Published: June 2019

Background:

Alternative splicing (AS) is essentially regulated by RNA‐binding proteins (RBPs) and is one of many deregulated cellular processes in Acute Myeloid Leukemia (AML). The alternative usage of exons substantially increases the coding potential of mRNAs and frequently leads to the expression of tumor‐promoting mRNA variants in cancer modulating apoptosis, proliferation, invasion and migration of cancer cells. This underlines the great plasticity of AS in cancer and justifies the need to elucidate the function of nuclear RBPs in specific cancer types. We found that the RNA helicase DDX17 is significantly more highly expressed in primary human AML specimens with high versus low Leukemia Stem Cell (LSC) frequency.

Aims:

In this study, we aimed to understand the functional role of DDX17 in normal hematopoietic stem and progenitor cells (HSPCs) and in AML cells. In particular, we sought to determine whether DDX17 exerts its function in the hematopoietic system primarily via AS or AS‐unrelated mechanisms.

Methods:

We performed knockdown (KD) experiments using small hairpin (sh) RNAs to determine whether DDX17 was essential for primary human cord blood CD34+ HSPCs and leukemia cells. Furthermore, mRFP tagged DDX17 was overexpressed through lentiviral transduction to track protein localization by immunofluorescence (IF) imaging. Lentivirally transduced HSPCs and leukemia cells were analyzed in vitro for proliferative output, viability, immunophenotype, and colony formation potential. RNA sequencing (RNA‐Seq) was used to determine how DDX17 KD affected gene expression and alternative exon usage. Pull down experiments combined with mass spectrometry revealed binding partners in presence and absence of chemotherapeutic agents. Finally, we performed homologous recombination (HR) and drug sensitivity assays to determine the role of DDX17 in DNA damage.

Results:

Knockdown of DDX17 induced DNA damage and severely impaired the function of healthy HSPCs and TP53 wild type AML cells. TP53 null cells maintained their proliferative capacity during short term in vitro expansion in absence of DDX17, but were significantly sensitized towards DNA damaging agents. In line, overexpression of DDX17 protected the cells against double strand break (DSB) inducing drugs. IF imaging revealed direct recruitment of DDX17 to DNA damage sites, while RNA‐Seq showed suppression of multiple DNA repair genes.

Summary/Conclusion:

RNA‐Seq, IF imaging, proteomics, and HR assays suggest an essential role of DDX17 in DNA repair via different mechanisms including direct recruitment to DNA damage sites, alternative splicing, and regulation of DNA repair genes on the transcriptional level.
PF298 LISOCABTAGENE MARALEUCEL TREATMENT OF PATIENTS WITH RELAPSED/REFRACTORY B‐CELL NON‐HODGKIN LYMPHOMA AND SECONDARY CENTRAL NERVOUS SYSTEM LYMPHOMA: INITIAL RESULTS FROM TRANSCEND NHL 001
Authors: Abramson J. , Palomba M.L. , Arnason J. , Lunning M. , Solomon S. , Farazi T. , Garcia J. , Xie B. , Newhall K. , Dehner C. , Siddiqi T.
Published: June 2019

Background:

No clinical studies have yet evaluated chimeric antigen receptor (CAR) T cell therapies in patients with relapsed/refractory (R/R) B‐cell non‐Hodgkin lymphoma (NHL) who have secondary central nervous system (CNS) lymphoma.

Aims:

To report data from patients with secondary CNS lymphoma receiving lisocabtagene maraleucel (liso‐cel; JCAR017), an investigational, anti‐CD19 CAR T cell product administered as a defined composition of CD4+/CD8+ CAR T cells, in the phase 1 TRANSCEND NHL 001 study.

Methods:

Eligible patients had confirmed B‐cell NHL with R/R disease after ≥2 prior lines of therapy. Patients with secondary CNS lymphoma were eligible as long as they also had systemic disease or, if it developed on study, patients could continue to receive liso‐cel. After lymphodepleting chemotherapy, liso‐cel was administered at 1 of 2 dose levels; 50 × 106 total CAR+ T cells (dose level 1) or 100 × 106 total CAR+ T cells (dose level 2). Efficacy was evaluated per the Lugano criteria. Patients achieving a complete response could be retreated with liso‐cel upon progressive disease.

Results:

At the data cutoff, 9 patients with secondary CNS lymphoma at initial treatment (n = 6), retreatment (n = 2), or cycle 2 (n = 1) received liso‐cel. Four patients were treated at dose level 1 and 5 at dose level 2. The median (range) age was 60 (47–73) years and the number of prior lines of therapy was 3 (2–7). Median (range) time to peak CAR+ T cell expansion was 12.5 (7–112) days. One of 9 patients had grade 2 cytokine release syndrome (CRS) and 1 of 9 patients had a neurological event (NE; grade 3 decreased level of consciousness). No retreatment patients had CRS or NE, however, 1 retreatment patient had an NE of grade 2 temporal edema during the initial treatment with liso‐cel. Five patients received prophylactic levetiracetam. One patient received corticosteroids and tocilizumab. Other toxicities were predominantly cytopenias. There were no treatment‐related deaths. Four patients responded to liso‐cel; all with a best response of complete response, of which 2 are ongoing at 270 and 545 days post‐liso‐cel. All 4 responses occurred after initial liso‐cel treatment; no retreated patients responded.

Summary/Conclusion:

In the ongoing TRANSCEND NHL 001 study, liso‐cel continues to demonstrate the ability to be safely delivered to patients with R/R B‐cell NHL, including those with secondary CNS lymphoma, a population of patients with a highly unmet medical need. No excess NE was noted in this population. This cohort continues to be evaluated.
PF299 EARLY RESPONSE OBSERVATION OF PEDIATRIC PATIENTS WITH BURKITT'S LYMPHOMA TREATED WITH CHIMERIC ANTIGEN RECEPTOR (CAR) T‐CELLS
Authors: Zhang W.‐Q. , Hu B. , Jing L. , Yang J. , Du J. , Wang S. , Ren Y.‐L. , Liu Y. , Zhang Y.‐H.
Published: June 2019

Background:

Pediatric patients with relapsed or refractory Burkitt's lymphoma (BL) present as a major challenge due to their poor outcomes despite use of high‐intensity chemotherapy and rituximab. CAR‐T has shown efficacy in treating refractory or relapsed leukemia and non‐Hodgkin lymphoma (NHL) in adult patients.

Aims:

To assess the safety and efficacy of CAR‐T in the treatment of refractory or relapsed BL in pediatric patients.

Methods:

Our trial (ChiCTR18000144) enrolled and treated 5 patients with BL. The inclusion criteria includes: 1) The patient who didn’t get complete remission(CR)after at least 5 courses of standard chemotherapy plus CD20 mAb; and 2) The patient who didn’t get partial remission (PR) or showing progression after 2 courses of second‐line chemotherapy; and 3) High expression of at least one B cells markers (CD19/CD20/CD22) examined by immunohistochemistry. The highest expressed marker was chosen as first round CAR‐T target. The therapeutic doses of CAR‐T range from

1∗10^6 to 3∗10^6 per kilogram of body weight each course. The effects and adverse events of CART cells after infusion were observed. The change of CAR‐T cells and cytokines were measured weekly. Tumor response was evaluated at day 30 and day 60 post infusion and every two months afterwards.

Results:

Five patients aged 6 to 10 years with a median age of 8 years, were enrolled, including 4 male (80%) and 1 female (20%). St Jude staging at the time of diagnosis: 3 cases (60%) in stage III, 2 cases (40%) in stage IV, 1 case of CNS invasion (20%), 1 case of bone marrow invasion (20%) and none in stage I or stage II.

Five patients were treated with 1 to 3 rounds of CAR‐T. The overall response rate (OR) was 100%. Three cases (60%) achieved CR by one round of CD19 CAR‐T cell therapy, while two cases (40%) achieved PR with CD19 CAR‐T treatment and achieved CR after subsequent treatment with CD22 or CD22 and CD20 CAR‐T.

During the treatment, fever, gastro‐intestine discomfort, rash, capillary leak syndrome, Immunopneumonia, tremor, hypotension, Immunocytopenia abnormal coagulation, liver dysfunction, and other toxic effects occurred. One patient (20%) was diagnosed of cytokine‐release syndrome(CRS) grading I and the other four grading III. All cases recoverd fully after active symptomatic and supportive treatment (including application of glucocorticoids in CRS grading III). There were no death events.

The last follow‐up was January 30, 2019. The longest follow‐up duration was 382 days, the shortest 62 days, and the median was 249 days. Currently, all patients were in event‐free survival.

Summary/Conclusion:

CD19/CD20/CD22‐CAR‐T therapy shows robust efficacy in pediatric patients with refractory and relapsed BL and the toxic and adverse effects are moderate and can be well controlled.
PF322 APPROACH TO DOUBLE HIT LYMPHOMA (DHL) DIAGNOSIS AND TREATMENT IN EUROPE – A CROSS‐SECTION STUDY OF EHA LYMPHOMA WORKING PARTY (EHA LYG)
Authors: Aurer I. , Kersten M.J. , Dreyling M. , Federico M.
Published: June 2019

Background:

DHL is a new lymphoma entity defined in the WHO 2018 classification. Diagnosis of DHL requires the use of FISH for MYC, BCL2 and BCL6, a technic that is not routinely used in the diagnostics of diffuse large‐B cell lymphomas (DLBCL). Treatment recommendations for DHL differ from those of DLBCL, but the level of evidence for them is low.

Aims:

EHA LYG performed this study to identify common practices in DHL diagnosis and treatment in different European countries.

Methods:

Lymphoma experts from different European countries were contacted and asked to fill out a questionnaire regarding recommendations and common practices in diagnosis and treatment of suspected DHL and DLBCL.

Results:

Almost half of European countries do not have national recommendations for DHL screening. Routine FISH screening is recommended in less than 20%, approximately 20% recommend screening all double‐expressors irrespective of cell of origin (COO), and 10% only GC‐type double‐expressors. COO is universally defined according to the Hans algorithm. MYC cut‐off values for defining double‐expressors are mostly 40% (but sometimes 70%), for BCL2 between 40% and 60% and for BCL6 60%. In one third of countries is screening recommended also in cases with clinical high‐risk features. FISH for DHL is routinely available to all DLBCL patients in approximately 40% of countries, in another 40% only to patients treated at referral hospitals, and not available at all in 20%. Only rarely is the availability of testing dependent on the type of insurance. Approximately a third of countries have nation‐wide recommendations for treating DHL. However, most patients with diagnosed DHL who are able to tolerate more aggressive therapies receive regimens different from R‐CHOP, mostly DA‐R‐EPOCH and Burkitt lymphoma‐type regimens, less frequently R‐CHOEP14. In approximately one third of the countries are alternative regimens used only in referral centers.

Summary/Conclusion:

The responses to our questionnaire suggest that current practices in recognizing DHL in Europe, even in relatively rich countries, result in missing of a substantial number of patients, especially in those treated outside of referral centers and with lower‐risk features. This fact must be taken into account when describing the natural history of DHL. Most patients diagnosed with DHL receive more aggressive treatment than those with DLBCL.
PF372 INHIBITION OF BROMODOMAIN AND EXTRA‐TERMINAL (BET) PROTEINS INCREASES SENSITIVITY TO VENETOCLAX IN CHRONIC LYMPHOCYTIC LEUKEMIA
Authors: Carrà G. , Lingua M.F. , Maffeo B. , Guerrasio A. , saglio G. , Taulli R. , Morotti A.
Published: June 2019

Background:

The development of drugs able to target BTK, PI3k‐delta and Bcl2 has dramatically improved Chronic Lymphocytic Leukemia therapies. However, resistant cases to these drugs have already been reported due to non‐recurrent changes in oncogenic pathways and genes expression signatures.

Aims:

In this study, we investigated thecooperative role of Venetoclax and the JQ1 inhibitor in order to prevent and eventually overcome the development of resistance to Bcl‐2 inhibitor.

Methods:

MEC‐1/EHEB cells lines and primary CD19 positive lymphocytes from CLL patients were treated with BET inhibitor JQ1 and venetoclax. Cell lines were screened for cell viability upon treatment with Venetoclax, JQ1and the combination of the two. Responses were measured as apoptosis induction (Annexin V), cell growth arrest and cell‐cycle accumulation. Furthermore, Western Blot analysis RT‐PCR and Immunofluorescence were used to determine expression of Brd members and BCL‐2 pathway components.

Results:

We demonstrated that treatment with BCL2 inhibitor is associated with a rapid emergence of resistant clones. Compared with each agent alone, cotreatment with JQ1 and venetoclax improved apoptosis in CLL cell lines and of the in vitro isolate primary CD19+ lymphocytes. Additionally, cotreatment could prevent the development of resistant clones. Lastly, JQ1 is also able to restore sensitiveness to venetoclax in venetoclax‐resistant CLL cell lines.

Summary/Conclusion:

Our findings indicated that cotreatment with JQ1 and BCL2 inhibitor synergistically induced apoptosis and eventually, prevent the development of resistant clones. Further knowledge is key to development of combinatorial treatment strategies with this two drugs.
PF377 ACALABRUTINIB WITH OBINUTUZUMAB IN TREATMENT‐NAIVE AND RELAPSED/REFRACTORY CHRONIC LYMPHOCYTIC LEUKEMIA: 3‐YEAR FOLLOW‐UP
Authors: Woyach J.A. , Rogers K. , Bhat S. , Blachly J. , Jianfar M. , Frigault M.M. , Hamdy A. , Izumi R. , Munugalavadla V. , Quah C. , Wang M.H. , Byrd J.C.
Published: June 2019

Background:

Acalabrutinib is a highly selective, potent, covalent Bruton tyrosine kinase inhibitor.

Aims:

This Phase 1b/2 trial evaluated acalabrutinib with the CD20 antibody obinutuzumab in patients with treatment‐naive or relapsed/refractory chronic lymphocytic leukemia (CLL).

Methods:

Patients with treatment‐naive or relapsed/refractory (≥1 prior therapy) CLL were eligible. In 28‐day cycles, acalabrutinib was given orally at 100 mg twice daily or 200 mg once daily (n = 15; all switched to 100 mg twice daily) until progressive disease; obinutuzumab was given in standard fashion for 6 cycles starting with Cycle 2. The primary endpoints were overall response rate (ORR) and safety. Minimal residual disease (MRD) was assessed using flow cytometry (sensitivity 10–4).

Results:

Nineteen treatment‐naive and 26 relapsed/refractory patients were treated. The median age of all patients was 61 years (range, 42 to 76). Patient characteristics, disposition, efficacy and MRD are in the Table. Common adverse events (any grade) were upper respiratory tract infection (71%), increased weight (71%), maculopapular rash (67%), cough (64%), diarrhea (62%), headache (56%), nausea (53%), arthralgia (51%), and dizziness (47%). Common Grade 3/4 adverse events were decreased neutrophil count (24%), syncope (11%), decreased platelet count, increased weight and cellulitis (9% each). There were 2 (4%) Grade 3 bleeding events (hematuria, muscle hemorrhage) and 1 (2%) Grade 3 atrial fibrillation event.

Summary/Conclusion:

Acalabrutinib plus obinutuzumab was well tolerated and yielded high response rates that were durable and deepened over time in treatment‐naive and relapsed/refractory patients with CLL.

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PF392 MONOCLONAL GAMMOPATHY AND HYPOGAMMAGLOBULINEMIA AS INDEPENDENT PROGNOSTIC FACTORS IN PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA: A RETROSPECTIVE MULTICENTRIC EXPERIENCE
Authors: Corbingi A. , Innocenti I. , Tomasso A. , Pasquale R. , Visentin A. , Varettoni M. , Autore F. , Morelli F. , Trentin L. , Reda G. , Laurenti L.
Published: June 2019

Background:

Chronic Lymphocytic Leukemia (CLL) is an indolent B‐cell lymphoproliferative disorder. Several prognostic factors such as IGHV mutation status and chromosomal aberrations as trisomy 12, del11q, del13q or del17p have been detected so far. More recent genetic mutations such as BIRC3, SF3B1, NOTCH1 and TP53 stratifies even further the prognosis and outcome in CLL patients (pts). However all data above reported, because of expensive techniques and experience typical of big laboratories, are not available to all medical centers while data concerning the presence of hypogammaglobulinemia, IgM or IgG monoclonal gammopathy are easily achievable, even though the prevalence and the impact on natural history of CLL pts is controversial and contradictory.

Aims:

The aim of the study is to evaluate the prevalence and the outcome of monoclonal IgM/CLL, IgG/CLL and hypogammaglobulinemia compared with CLL pts with normal immunoglobulin (Ig) levels.

Methods:

We collected from four different Italian centers 1359 pts diagnosed with typical CLL from 1987 to 2017 with a baseline assessment of serum Ig, immunofixation, chromosomal aberrations and clinical features; evaluating their impact on time to progression (TTP), time to treatment (TTT) and overall survival (OS).

Results:

Once assessment was made, pts included in our study who met eligibility criteria were 1359. Median age was similar in all the groups and data gathered from the four centers showed an overlapping rate about prevalence and time‐dependent‐parameters in CLL pts and their related subclasses.The overall prevalence of monoclonal gammopathy is 14% of whom 128 pts (9%) with IgG/CLL, 68 pts (5%) with IgM/CLL. Hypogammaglobulinemia was detected in 172 pts (13%) while 991 pts (73%) had no evidence of paraprotein. Among the groups, serum IgG paraprotein showed a correlation with a poorer and shorter overall survival (OS) and an inferior clinical outcome.

Summary/Conclusion:

Our study provided data about prevalence and prognosis of monoclonal gammopaty and hypogammaglobulinemia in CLL patients. Serum Ig paraprotein was present in 14% of all CLL pts with a worst outcome in IgG/CLL group. Thus, making of serum Ig, a simple data to obtain, useful to assess CLL prognosis.

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