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Introduction: Current studies demonstrate an involvement of germline predispositions in the development of approximately 5% of childhood leukemias (Zhang J et al.,N Engl J Med, 2015), although their actual contribution is believed to be much higher. Being able to understand tumor evolution starting from a predisposed cell, opens up a new avenue in the form of disease prevention rather than treatment. Here, we present a novel finding of a double hit - one pathway scenario, in which a rare (MAF Methods: WES was carried out to identify predisposing germline variants. The cooperative functionality of the di-genic candidate mutations was experimentally tested using BaF3 oncogenic transformation assays, western blot and cell cycle analyses. In addition, we used structural homology calculations to visualize the cooperative impact of both mutations. Results: Utilizing Trio-calling based on WES we identified two concomitant germline SNVs in the JAK2/STAT3 pathway in a boy affected with BCP-ALL. The JAK2 variant, rs139504737, leads to an amino acid substitution from Glycine to Serine (p.G571S) (MAFG), causing an exchange of Lysine to Arginine (p.K370R). Structural modeling of STAT3 K370R showed that K370, which is an important site for acetylation, is located in a loop adjacent to the DNA binding site of STAT3. The substitution of Lys to Arg at position 370 leads to a strengthening of the beta-sheet due to simultaneous interaction of Arg with both E455 and E442. Moreover, in contrast to Lys, Arg cannot be acetylated, leading to a constitutively non-acetylated form of STAT3. JAK2 G571S on the other hand has a unique position affecting amino acid 571, which lies adjacent to the Y570 residue that downregulates kinase activity via autophosphorylation, indicating a potential functional mechanism of the G571S mutation by inhibiting Y570-directed negative feedback. To assess the cooperative oncogenic transforming potential of both variants, BaF3 depletion assays were carried out. In BaF3/CRLF2-IL-7Rwt cells, JAK2 G571S conferred IL-3 independent growth at similar rates as the well-known oncogenic JAK2 V617F mutation. Moreover, the combination of both JAK2 G571S and STAT3 K370R further increased the growth advantage significantly starting 2 days after IL-3 withdrawal (p=0.0226). Western Blot analyses revealed increased pSTAT5 levels, as well as high pSTAT-3 levels in the double mutant cells. We further observed that STAT3 K370R alone changed the phenotype of the in-vitro culture, with an accumulation of enlarged BaF3 cells. Surprisingly, this phenotype was reversed in cells expressing both STAT3 K370R and JAK2 G571S. Cell cycle analysis showed a significant increase of aneuploid cells (G2/M) (p=0.0009), while the G-1 phase was significantly decreased (p=0.0031) in STAT3 K370R expressing BaF3 cells compared to STAT3 WT cells. Again, this phenotype was drastically reduced in cells transfected with both mutations simultaneously (G1 phase p=0.0026; G2/M p=0.0032). Western Blot analyses confirmed increased p-CDC-2, p-Cyclin/Cyclin-B1, and Cyclin-A2 levels in BaF3 cells harboring both variants, suggesting that the cell cycle arrest observed in cells expressing STAT3 K370R is rescued by JAK2 G571S expression by enabling re-entering of the M-Phase. The link between the oncogenic capacity of JAK2 G571S and leukemia could further be strengthened by the identification of an additional B-precursor ALL patient from an independent family harboring the same JAK2 G571S germline mutation. The patient belongs to a previously described DS-ALL cohort (Bercovich D. et al., Lancet, 2008). In this patient with Down syndrome, we did neither identify the STAT3 K370R mutation nor CRLF2 activation. Thus, we hypothesize that the germline JAK2 G571S in combination with the constitutional trisomy 21 act in synergy. Conclusion: Taken together, we present evidence of an oncogenic potential of germline JAK2 G571S, which is significantly increased through additional expression of STAT3 K370R in a double-hit one-pathway scenario. Disclosures No relevant conflicts of interest to declare.

Double hit B-cell lymphomas with concurrent MYC and BCL2 rearrangements are well described and are associated with a rapidly progressive clinical course and dismal prognosis. Cases of double hit myeloma, however, are rare, and there are no reported cases of double hit plasma cell leukemia (PCL).

Double-hit lymphomas, usually refer to the MYC/BCL2 lymphomas, and are characterized by MYC 8q rearrangement and BCL2-IgH translocation in the same cells. They mainly comprise of a subset of the new entity “B cell lymphomas unclassifiable with features intermediate between diffuse large B cell lymphomas (DLBCL) and Burkitt lymphoma” included in the World Health Organization 2008 classification. In addition, they also include a small subset of other DLBCL. They have been reported to have worse prognosis than either Burkitt or DLBCL, with which they share overlapping morphological and immunophenotypical features. These cases usually present de novo and have been reported in the older age group (Snuderl et al. in Am J Surg Pathol 34(3):327–340, 2010; Le Gouill et al. in Haematologica 92(10):1335–1342, 2007; Macpherson et al. in J Clin Oncol 17(5):1558–1567, 1999; Johnson et al. in Blood 114(11):2273–2279, 2009; Bertrand et al. in Leuk Off J Leuk Soc Am Leuk Res Fund UK 21(3):515–523, 2007). Here, we describe an unusual simultaneous presentation of MYC/BCL2 double-hit lymphoma with low-grade follicular lymphoma, occurring at two different sites, in a young patient. This has not been reported to the best of our knowledge.

Although a lot of data are available in the literature on the recrystallization behavior of plain carbon and microalloyed steels, comparisons are often difficult to make due to the effect of different experimental techniques and the type of tests used to obtain these data. Few systematic comparisons can be found in the literature to correlate the different techniques and methods. A previous paper by the present authors concentrates on the analysis of torsion methods. The present study focuses on the recrystallization kinetics determined by using double hit and stress relaxation tests in plane strain compression mode. The specificity of the test and the strain distribution across the section has been incorporated into the analysis, by means of finite element methods. A good correlation has been obtained between the kinetics for stress relaxation and double hit tests. However, some rules should be respected in order to determine the true recrystallization/softening kinetics comparable with those obtained for other deformation modes like torsion.

Different recrystallization models are now available in the literature for plain carbon and microalloyed steels. However, the attempts made to unify these models have been unsuccessful due to the considerable scatter in experimental results that support the corresponding equations. In fact, kinetics results can be separated into different groups depending on the testing procedure. In order to cope with this problem and improve the reliability of the different models, the different methods that are currently applied at laboratory need to be re-analyzed. The present work concentrates on the determination of recrystallization/softening kinetics of austenite from torsion tests. To this purpose the softening results obtained by the application of double hit torsion tests to two Nb microalloyed steels and the fraction recrystallized as determined by metallography have been subjected to a direct and an inverse analysis that takes into account the specificity of the torsiontest.

Multiple genes are associated with thoracic aortic aneurysm (TAA) formation, including FBN1, TGFBR1&2, COL3A1 & 5A2, and MYH11. Given the population incidence of mutations in these genes we hypothesise that it is likely that ‘double hit’ mutations occur, which may result in an altered clinical phenotype when compared with the single gene mutation. Genomic DNA analysis of 19 genes implicated in TAA was performed on 12 patients with inherited TAA disease. Of the patients analysed, one patient (P1) was identified as having a mutation in both MYH11 (c.2517G.C (pW839C)) and FBN1 (splice donor site c.4210+1G>A). Another patient (P2) was identified as having both COL5A2 (c.3794A>G (pD1265G)) and FBN1 (c.5861T>G (p.F1954C)). All mutations were predicted by PolyPhen to be deleterious. P1 has Marfan Syndrome (MFS), caused by the FBN1 mutation, and has severe aortic dilatation while his mother (M1), who also has MFS but does not have the MHY11 mutation, developed mild aortic dilatation, suggesting that the MHY11 mutation in P1 has resulted in a more aggressive phenotype. P2, siblings S1 and S2 and father (F2) all have relatively less severe MFS with progressive moderate dilatation of the aorta. Thus, the phenotype of pedigree 2 is similar for all affected members, but only P2 has a double hit mutation, suggesting that the presence of the COL5A2 mutation in P2 has not altered the clinical phenotype. This is the first instance where double hit mutations have been described in inherited thoracic aortic aneurysm but the effect of multiple mutations on phenotype appears to be variable. Mutation analysis for individuals with inherited thoracic aortic aneurysm should consider the possibility of multiple gene culprits.

This article examines the psychological impact of the online dating romance scam. Unlike other mass-marketing fraud victims, these victims experienced a ‘double hit’ of the scam: a financial loss and the loss of a relationship. For most, the loss of the relationship was more upsetting than their financial losses (many described the loss of the relationship as a ‘death’). Some described their experience as traumatic and all were affected negatively by the crime. Most victims had not found ways to cope given the lack of understanding from family and friends. Denial (e.g. not accepting the scam was real or not being able to separate the fake identity with the criminal) was identified as an ineffective means of coping, leaving the victim vulnerable to a second wave of the scam. Suggestions are made as to how to change policy with regards to law enforcement to deal with this crime.

Purpose of review Mature B-cell lymphomas bearing concurrent chromosomal rearrangement of MYC/8q24 and BCL2/18q21 are associated with an aggressive clinical course and resistance to conventional chemotherapy. This review summarizes the recent literature regarding the clinical and pathologic features of double-hit lymphomas and outlines current questions about the most accurate and inclusive definition of the disease. Recent findings Comprehensive evaluation of large series of aggressive mature B-cell neoplasms reveals recurrent chromosomal aberrations in the majority of cases. A subset of these lymphomas harbors multiple rearrangements, including MYC/8q24 in combination with BCL2/18q21 and/or BCL6/3q27, and displays a particularly aggressive clinical course. Recent data suggest that consideration of additional features, such as copy number alteration, quantitative protein expression, and biologic pathway activation may be important in deriving a more accurate definition of double-hit lymphoma. Despite the poor prognosis associated with this subset of lymphomas, there remains no evidence for a risk-adapted treatment strategy and no clinical, pathologic, or genetic factors that predict response to therapy. Summary Double-hit lymphoma remains an incompletely characterized disease entity. Large, multicenter studies are needed to define relevant clinical, genetic, and pathologic variables and to characterize appropriate risk-adapted treatment strategies.

The sovereign debt crisis affecting several eurozone countries is a threat to Europe's financial stability and has had significant international repercussions. The fear of a default within the eurozone has spread from Greece to other states. The various summits that have taken place on the subject have provided only partial and Insufficient responses to the problems confronted. For several months now, it has appeared that one of the main impediments to resolving the crisis has been the massive exposure of the financial sector to the debt of eurozone states. A fall in the value of this debt, owing to investor distrust, may lead to banks requiring financial injections to protect their capital positions. As a result, additional funds might well be required from eurozone states. Such support might come from the states directly or come from them indirectly via the European Financial Stability Facility: a double hit for eurozone members. The crisis has made clear the fundamental imbalances in the economies of E...

In this issue of Cancer Cell, Diamantopoulou et al. uncover dual mechanisms of inhibiting YAP/TAZ by TIAM1 that oppose invasiveness of colorectal cancer cells: TIAM1 interacts with TAZ in the cytoplasm to promote TAZ degradation by the destruction complex, whereas it antagonizes binding of TAZ/YAP to TEAD in the nucleus.

We report on a test of a straw tube detector design having several anode (sense) wires inside a straw tube. The anode wires form a circle inside the tube and are read out independently. This design could solve several shortcomings of the traditional single wire straw tube design such as double hit capability and stereo configuration.

Thin slab casting and direct rolling technologies became important for hot strip production to increase the productivity, to lower the investment costs and to increase the energy efficiency. The main objective of our investigation is to gain a better understanding of the microstructural evolution starting from solidification through to the final hot rolled sheet. Small ingots were cast and direct rolled and reheated and rolled, using a thermomechanical testing machine. The differences in the recrystallization kinetics during both production routes were measured using double hit hot compression tests. In addition, multipass deformation trials were performed to reveal differences in the resulting microstructure and the grain size homogeneity. It was found, that the coarse initial microstructure before rolling become negligible after three rolling passes. This means that despite the simple and economical direct rolling similar mechanical strip properties can be obtained in comparison to conventional strip production routes.

Heterozygous mutations in the PINK1 gene are considered a susceptibility factor to develop early-onset Parkinson's disease (PD), as supported by dopamine hypometabolism in asymptomatic mutation carriers and subtle alterations of dopamine-dependent striatal synaptic plasticity in heterozygous PINK1 knockout (PINK1(+/-)) mice. The aim of the present study was to investigate whether exposure to low-dose rotenone of heterozygous PINK1(+/-) mice, compared to their wild-type PINK1(+/+) littermates, could impact on dopamine-dependent striatal synaptic plasticity, in the absence of apparent structural alterations. Mice were exposed to a range of concentrations of rotenone (0.01-1mg/kg). Chronic treatment with concentrations of rotenone up to 0.8mg/kg did not cause manifest neuronal loss or changes in ATP levels both in the striatum or substantia nigra of PINK1(+/-) and PINK1(+/+) mice. Moreover, rotenone (up to 0.8mg/kg) treatment did not induce mislocalization of the mitochondrial membrane protein Tom20 and release of cytochrome c in PINK1(+/-) striata. Accordingly, basic electrophysiological properties of nigral dopaminergic and striatal medium spiny neurons (MSNs) were normal. Despite the lack of gross alterations in neuronal viability in chronically-treated PINK1(+/-), a complete loss of both long-term depression (LTD) and long-term potentiation (LTP) was recorded in MSNs from PINK1(+/-) mice treated with a low rotenone (0.1mg/kg) concentration. Even lower concentrations (0.01mg/kg) blocked LTP induction in heterozygous PINK1(+/-) MSNs compared to PINK1(+/+) mice. Of interest, chronic pretreatment with the antioxidants alpha-tocopherol and Trolox, a water-soluble analog of vitamin E and powerful antioxidant, rescued synaptic plasticity impairment, confirming that, at the doses we utilized, rotenone did not induce irreversible alterations. In this model, chronic exposure to low-doses of rotenone was not sufficient to alter mitochondrial integrity and ATP production, but profoundly impaired the expression of long-term plasticity at corticostriatal synapses in PINK1 heterozygous knockout mice, suggesting that disruption of synaptic plasticity may represent an early feature of a pre-manifesting state of the disease, and a potential tool to test novel neuroprotective agents.

e19530Background: Double-hit lymphomas (DHLs) are high-grade diffuse large B-cell lymphomas with concurrent translocations involving myc and bcl-2 and/or bcl-6. A patient with DHL often has advance...

For the investigation of retained strains during hot forming, which are related to the dislocation structure, single and double hit compression tests were carried out at different temperatures and strain rates for a stainless steel. Using microhardness measurements the retained strains after the first and second pass were investigated as a function of the amount of deformation, temperatures as well as strain rates and dwell durations. In general, the retained strain decreases with increasing dwell durations in both the deformed and recrystallized grains, respectively. At a given total amount of deformation in a double hit compression, the retained strains for the as deformed unrecrystallized grains are reduced for a lower deformation in the first hit. For the recrystallized grains the retained strain increases, when comparing double hit with single hit compression.

Double hit compression tests were performed on an ultra low carbon Si-Al alloyed steel. The true strain in both deformation steps was kept constant (0.2), while varying the strain rate (1 s -1 , 10 s -1 and 30 s -1 ), the temperature (1,050°C-austenite phase, 950°C-intercritical region and 850°C-ferritic phase) and the interpass time (0.5; 1; 5 and 50 s). The temperatures used are within the industrial range applied in the finishing rolling for this steel. The analysis of the flow curves obtained allowed the evaluation of the static softening characteristics of the steel. It was shown that the steel has a similar softening condition between passes at the three temperatures studied, with the softening fraction increasing with the increasing time and strain rate. The characteristics of the softening curves and microstructural features indicate that at the two higher temperatures (austenite phase and upper part of the intercritical region) the softening process is governed by recrystallization, with the nucleation and growth of subgrains. At the lower temperature, however, the softening is mainly due to strain induced boundary movement (SIBM) and recovery.

Double-hit compression tests were carried out at different temperatures and strain rates for a nickel based alloy and a stainless steel. Using microhardness measurements the retained strains after the first and second pass were investigated as a function of the amount of deformation, temperatures as well as strain rates and dwell durations. In general, the retained strain decreases with increasing dwell durations. It is shown that at a given total amount of deformation, the retained strain is reduced for the as deformed grains that have not been recrystallized yet, but increased for the recrystallized grains, when comparing double hit with single hit compression tests.

Static recrystallisation kinetics during hot deformation of two microalloyed steels (C–Ti–V and C–Ti–Nb) has been quantified. Double hit compression test using a Gleeble 1500 thermomechanical simulator was conducted to determine the recrystallisation kinetics. The kinetics of static recrystallisation was found to be more sluggish in the case of microalloyed steels with niobium additions as compared with other grades of microalloyed steels. The rate of recrystallisation increases with increasing temperature, strain and strain rate. The results are compared with other grades of microalloyed steels, mainly with Ti, Ni and Cu additions, from literature. For niobium containing microalloyed steels, a higher temperature is required for recrystallisation as compared with other microalloyed steels.

By means of hot compression single and double hit experiments, the kinetics of dynamic and static recrystallization in hot-rolled Si-Mn TRIP steel was studied, and the emphasis was put on the influence of high silicon content. The results show that the calculated parameters are consistent with the experimental ones, and addition of silicon retards both dynamic and static recrystallization as well as increases the flow stress of austenite, and the non-recrystallization zone can be enlarged by increasing the silicon contents.

We hypothesized that the double hit conferred by sex and diagnosis increases the risk for internalizing disorders in adolescent females with autism spectrum disorder (ASD). In a sample of 32 adolescents with ASD and 32 controls, we examined the effects of sex, diagnostic factors, and developmental stages on depression and anxiety. A 3-way interaction revealed that females with ASD exhibited greater depressive symptoms than males with ASD and female controls particularly during early adolescence; therefore, females with ASD might have a unique combination of genetic, hormonal, and psychosocial vulnerabilities that heighten their risk for depression during early adolescence. Additionally, the ASD group reported high levels of separation anxiety and panic in late adolescence, possibly indicating atypical development of independence.

Sickle cell trait (SCT) has been associated with exertional collapse (ECAST) and exercise-related sudden death in athletes and military warfighters. The mechanisms underlying ECAST remain controversial in the sports medicine community. Multiple case presentations and anecdotal reports postulate the role of extraordinary exercise intensity, but other risk factors including dehydration, heat, previous exertional rhabdomyolysis, genetic cofactors, and dietary supplements have been cited as potential contributors. Others have hypothesized some of the aforementioned factors combining in a "perfect storm" to trigger ECAST with a resultant potentially fatal "metabolic crisis." This case report provides a brief review of SCT as it pertains to exercise in warfighters and athletes, identifies known and postulated risk factors associated with ECAST, and introduces the potential mechanistic role of the "double hit" as a contributor to ECAST.

By using isothermal double hit compression tests and applying the 2% offset method, a new model was developed to predict the microstructural evolution of Nb-bearing steels at temperatures above and below the start temperature of strain-induced precipitation (T P ). The T P was developed as a function of true strain, initial austenite grain size and the Nb content. The activation energy of static recrystallization (Q rex ) was expressed as a function of the content of different alloy elements. It was found that Nb played the most important role in increasing the value of Q rex The microstructural observations and measurements confirmed the validity of the model developed in the present investigation.

We report the first case of a neonate with concurrent Chiari II malformation and achondroplasia. Although rare, both these conditions contribute to several deleterious anatomical changes at the cervicomedullary junction and thus predispose to acute hydrocephalus. Although our patient was initially asymptomatic, hydrocephalus ensued several weeks after birth and required cerebral spinal fluid diversion. We discuss the potential links between the two conditions, the pathophysiology, and the important clinical implications for the management of the increased risk of hydrocephalus.

The mechanical behaviour of a Nb-microalloyed steel under non monotonie conditions has been studied using double hit hot torsion tests involving changes in the twist direction. For strain reversal conditions, a clear mechanical transient has been observed. This transient has been characterized by a steep hardening region just after reversal which takes place up to a stress level near the one reached before reversing, then a region showing a low strain hardening rate and finally, a region extending up to the convergence of monotonie and reversal behaviours. Testing temperatures and strain rates have ranged from 950°C to 1200°C and from 0.01s-1 to 0.1s-1, respectively. Applied prestrains before reversal have varied from 0.03 to 0.25 in order to avoid the occurrence of dynamic recrystallization during previous strain. Lastly, some relationships have been established between the deformation conditions and the parameters defining the transient. These parameters depend mainly on the prestrain.Se ha estudiado el comportamiento mecánico de un acero microaleado con niobio bajo condiciones no monótonas de deformación mediante ensayos de torsión en caliente de doble pasada con cambio de sentido de giro entre la primera y la segunda de ellas. Se ha observado que la inversión de la deformación produce un transitorio en las curvas de fluencia caracterizado por un endurecimiento rápido hasta alcanzar un nivel de tensión menor al que se tenía, justo, antes de aplicar el cambio en el sentido de giro, una etapa de endurecimiento nulo y, finalmente, una región en la que el comportamiento mecánico durante la inversión tiende progresivamente al comportamiento mecánico monótono. Las temperaturas de ensayo se han variado entre 950 °C y 1.200 °C y las velocidades de deformación entre 0,01 y 0,1 s-1 Las deformaciones previas aplicadas antes de la reversión han estado comprendidas entre 0,03 y 0,25, evitándose la interacción con la recristalización dinámica. Finalmente, se han establecido diferentes relaciones entre las condiciones de deformación y los parámetros que definen el transitorio, observándose que estos últimos dependen principalmente de la deformación previa a la inversión.

During hot forging, the billet is heated to an elevated temperature, forged to the required shape and cooled to room temperature. Most of the hot forged parts produced, meet the specified geometric tolerances, but some do not. Some of the defective parts can be rectified by machining, others are rejected. Restoring a hot forged part demands extra production cost. Defective parts can be attributed to process variations or uncertainties that occur during production. Such manufacturing anomalies in the metal forming industry can induce heavy losses (e.g. premature failure of equipment, under-fill, folds in part and double hit) during mass production. The objectives of this research are, determining the uncertainties that cause defective parts during hot forging and illustrate a method to determine parameters that control the trade-off between rejected parts and rectifiable parts. This information will aid in reducing production costs of the forgings. The uncertainties are demonstrated in this research by simulations of the forging and cooling processes. Since, uncertainties in the field of metal forming have never been considered in the design process, quantifying the effects of the process uncertainties would be the first step towards reducing geometric variation and related production costs. Also, incorporating the process uncertainties into the forging and cooling process design leads to better quality parts.

Evolutionary pressure has selected individuals with traits that allow them to survive to reproduction, without consideration of the consequences for the post-child rearing years and old age. In the 21st century, society is populated increasingly by the elderly and with the falling birth rate and improved health care this trend is set to continue for the foreseeable future. To minimize the potential burden on health services one would hope that 'growing old gracefully' should also mean 'growing old healthily'. However, for too many the aging process is accompanied by increasing physical and mental frailty producing an elevated risk of physical and psychological stress in old age. Stress is a potent modulator of immune function, which in youth can be compensated for by the presence of an optimal immune response. In the elderly the immune response is blunted as a result of the decline in several components of the immune system (immune senescence) and a shifting to a chronic pro-inflammatory status (the so-called 'inflamm-aging' effect). We discuss here what is known of the effects of both stress and aging upon the innate immune system, focusing in particular upon the age-related alterations in the hypopituitary-adrenal axis. We propose a double hit model for age and stress in which the age-related increase in the cortisol/sulphated dehydroepiandrosterone ratio synergizes with elevated cortisol during stress to reduce immunity in the elderly significantly.

The metadynamic and static recrystallization behavior in hypereutectoid steel containing 1% carbon was determined by hot compression testing. Compression tests were performed using double hit schedules at temperatures between 900 to 1050°C, strain rates of 0.01 to 1 s−1 and recrystallization times of 0.1 to 500 s. The characteristics of static and metadynamic recrystallization are distinctly different. Results show that the metadynamic kinetics was twice as fast as the static kinetics. These data were used to generate equations to predict the kinetics of static and metadynamic recrystallization, as well as the evolution of grain size, after recrystallization., available, unclassified, unlimited

Introduction: Double Hit (DH) or Double Expresser (DE) DLBCL are aggressive Non-Hodgkin’s Lymphomas (NHLs) with translocation or over-expression of MYC and BCL-2 which are incurable with current therapies. MYC is a critical driver of oncogenic phenotypes and regulates expression of numerous genes including aurora kinases (AKs), which are validated drug targets. AKs are over-expressed in DLBCL and AK inhibition in two Phase-II trials of relapsed and refractory B/T-NHL resulted in 20%-30% overall response rate. Bruton’s tyrosine kinase (BTK) plays a central role in B-cell receptor (BCR) signaling and inhibitors of BTK have shown efficacy in B-NHL clinical trials. Targeting CD20 (rituximab) in DLBCL inhibits BCL-2 and induces apoptosis via caspase-3 activation. We hypothesized that DH/DE-DLBCL are amenable to a novel therapeutic strategy of AK, BTK and CD20 inhibition. Methods: Four DH/DE DLBCL cell lines were evaluated for viability by MTS; polyploidy, senescence and apoptosis by flow cytometry; DNA damage by TUNEL assay. Phospho-BCL-2, -AK and -BTK analysis by western. Safety and efficacy of drug combinations in a mouse xenograft model. Analysis of patient samples and mouse tumors by IHC. Results: DH/DE-DLBCL cell lines and patient samples displayed over-expression of MYC, BCL-2, BTK and AK-A proteins. Cell viability assays with alisertib alone showed ~30% cell death, while majority of remaining ~70% cells were polyploid and senescent at day-4 (a mechanism of treatment failure). These polyploid and senescent cells (PASC) showed increased levels of phospho-BTK indicating increased BCR signaling (a rationale for combination therapy). We show that alisertib + ibrutinib + rituximab significantly reduced PASCs leading to intrinsic cell death compared to single/double combinations. Dual inhibition of AK + BTK reduced phospho-ERK-1/2, -Bcl-6 and -MYC with increased DNA breaks indicated by up regulation of phospho-H2A-X, -Chk-2, and pro-apoptotic Noxa and Puma proteins. Triple therapy reduced Bcl-2, Bcl-xL and increased Noxa and Puma compromising the mitochondrial outer membrane potential, release of Cyt-C, activation of caspase-3 and induction apoptosis by PARP cleavage. A DLBCL SCID mouse xenograft model showed ibrutinib alone is inactive while alisertib + ibrutinib was additive with a tumor growth inhibition (TGI) rate of ~25%. TGI for ibrutinib + rituximab was ~50-60%. However, triple therapy showed a TGI of >90% and was well tolerated. Survival analysis: 67% of mice alive at day-89 with triple therapy versus 20% with ibrutinib + rituximab. Other treatments showed no survival past day-52. Conclusions: A novel triple therapy consisting of alisertib + ibrutinib + rituximab inhibits PASCs induced by AK inhibition in DH/DE-DLBCL leading to a significant anti-proliferative signal and intrinsic apoptosis. Our work provides a rationale for exploring these combinations in the clinic. Citation Format: Shariful Islam, Wenqing Qi, Carla Morales, Laurence Cooke, Catherine M. Spier, Daruka Mahadevan. Inhibition of polyploidy and senescence induces intrinsic apoptosis in double hit or double expresser DLBCL [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4057. doi:10.1158/1538-7445.AM2017-4057

7559 Background: Large B cell lymphomas with MYC, BCL2, and BCL6 rearrangements, designated as triple hit lymphoma (THL), are uncommon. Large series studies of THL are scant and studies comparing THL to different types of double hit lymphoma (DHL) are lacking. Methods: We studied the clinicopathologic features and prognosis of 33 patients with THL and compared them to 83 patients with MYC/BCL2 DHL and 13 patients with MYC/BCL6DHL. Results: There were 21 men and 12 women, with a median age of 63 years (range, 34-85). Six patients had a history of low-grade B cell lymphoma and 27 had de novo lymphoma. These tumors were classified histologically as: 21 DLBCL, 10 high grade B-cell lymphoma, one concurrent DLBCL and follicular lymphoma (FL), and one concurrent DLBCL and mantle cell lymphoma. Immunohistochemical analysis showed that these tumors were positive for CD10 (94%), BCL6 (80%), BCL2 (93%), and MYC (69%, 40% as cutoff). 62% of tumors (8/13) with available data showed coexpression of MYC and BCL2. Using the Hans algorithm, 30 of 33 (91%) tumors had a germinal center B cell like (GCB) immunophenotype. All 7 cases tested by conventional cytogenetics showed a complex karyotype. Although BCL2 was always translocated with IGH, BCL6 translocated to MYC in 2of 7 cases of THL. Twenty-nine patients had treatment information available and all received immune-chemotherapy induction, including 11 with R-CHOP, 14 with R-EPOCH, 3 with R-HyperCVAD, and one with RICE (patient had a history of FL). The clinicopathological features of THL including induction chemotherapy were very similar to both the MYC/BCL2 DHL and MYC/BCL6 DHL (all P>0.05). There was no significant difference in median overall survival (OS) between patients with de novo lymphoma and those with a history of low-grade lymphoma (P=0.99). The OS in THL patients was 17.9 months, similar to the OS (17.2 months) of patients with MYC/BCL6 DHL and those with MYC/BCL2DHL (19.9 months) (P=0.60). Conclusions: MYC/BCL2/BCL6 THL is an aggressive B cell lymphoma and >90% of cases have a GCB immunophenotype. THL patients usually have an aggressive clinical course and a poor prognosis, similar to patients with double hit lymphomas.

Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma histology, with 40% of patients cured with frontline therapy. Salvage chemotherapy followed by autologous hematopoietic cell transplant (HCT) remains the standard of care for relapsed or primary refractory patients who are chemosensitive. Autologous HCT in first remission is not recommended, as randomized trials have not shown a survival benefit. Despite evidence for a graft versus lymphoma effect, allogeneic HCT is often reserved for patients with DLBCL who have persistent marrow involvement, have failed autologous HCT or have primary refractory disease. Reduced intensity or non-myeloablative conditioning regimens carry a lower non-relapse mortality risk as compared to myeloablative conditioning but are associated with higher relapse. Patients with DLBCL with the dual translocations of BCL2 and MYC or "double hit" lymphoma carry a poor prognosis, and HCT is often offered as consolidation therapy. Further studies are needed to determine whether HCT can alter the natural history of this aggressive subtype.

The WHO classification of lymphomas allows for a group of diseases that have features intermediate between those of Burkitt lymphoma and diffuse large B-cell lymphoma. These are a diverse group of diseases whose genetics and clinical course are yet to be fully described. We report an unusual case of high grade B-cell lymphoma, intermediate between DLBCL and BL, lacking CD10 expression in which the chromosomal translocation t(3;8)(q27;q24) was found to be the sole chromosomal abnormality. FISH analysis demonstrated juxtaposition of theBCL6andMYCloci without obvious involvement of theIGHlocus, suggesting constitutiveMYCexpression due to promoter substitution. The patient responded to intensive chemotherapy and remains in remission two years after finishing therapy.