Search

There is a need to develop novel approaches to improve the balance between efficacy and toxicity for transcription factor–targeted therapies. In this study, we exploit context-dependent differences in RNA polymerase II processivity as an approach to improve the activity and limit the toxicity of the EWS-FLI1–targeted small molecule, mithramycin, for Ewing sarcoma. The clinical activity of mithramycin for Ewing sarcoma is limited by off-target liver toxicity that restricts the serum concentration to levels insufficient to inhibit EWS-FLI1. In this study, we perform an siRNA screen of the druggable genome followed by a matrix drug screen to identify mithramycin potentiators and a synergistic “class” effect with cyclin-dependent kinase 9 (CDK9) inhibitors. These CDK9 inhibitors enhanced the mithramycin-mediated suppression of the EWS-FLI1 transcriptional program leading to a shift in the IC50 and striking regressions of Ewing sarcoma xenografts. To determine whether these compounds may also be liver protective, we performed a qPCR screen of all known liver toxicity genes in HepG2 cells to identify mithramycin-driven transcriptional changes that contribute to the liver toxicity. Mithramycin induces expression of the BTG2 gene in HepG2 but not Ewing sarcoma cells, which leads to a liver-specific accumulation of reactive oxygen species (ROS). siRNA silencing of BTG2 rescues the induction of ROS and the cytotoxicity of mithramycin in these cells. Furthermore, CDK9 inhibition blocked the induction of BTG2 to limit cytotoxicity in HepG2, but not Ewing sarcoma cells. These studies provide the basis for a synergistic and less toxic EWS-FLI1–targeted combination therapy for Ewing sarcoma.

Purpose:The successful clinical translation of compounds that target specific oncogenic transcription factors will require an understanding of the mechanism of target suppression to optimize the dose and schedule of administration. We have previously shown trabectedin reverses the gene signature of the EWS-FLI1 transcription factor. In this report, we establish the mechanism of suppression and use it to justify the reevaluation of this drug in the clinic in patients with Ewing sarcoma.Experimental Design: We demonstrate a novel epigenetic mechanism of trabectedin using biochemical fractionation and chromatin immunoprecipitation sequencing. We link the effect to drug schedule and EWS-FLI1 downstream target expression using confocal microscopy, qPCR, Western blot analysis, and cell viability assays. Finally, we quantitate target suppression within the three-dimensional architecture of the tumor in vivo using 18F-FLT imaging.Results:Trabectedin evicts the SWI/SNF chromatin-remodeling complex from chromatin and redistributes EWS-FLI1 in the nucleus leading to a marked increase in H3K27me3 and H3K9me3 at EWS-FLI1 target genes. These effects only occur at high concentrations of trabectedin leading to suppression of EWS-FLI1 target genes and a loss of cell viability. In vivo, low-dose irinotecan is required to improve the magnitude, penetrance, and duration of target suppression in the three-dimensional architecture of the tumor leading to differentiation of the Ewing sarcoma xenograft into benign mesenchymal tissue.Conclusions:These data provide the justification to evaluate trabectedin in the clinic on a short infusion schedule in combination with low-dose irinotecan with 18F-FLT PET imaging in patients with Ewing sarcoma.

Healthy ageing involves degeneration of the neuromuscular system which impacts movement control and proprioception. Yet the relationship between these sensory and motor deficits in upper limb reaching has not been examined in detail. Recently, we reported that age-related proprioceptive deficits were unrelated to accuracy in rapid arm movements, but whether this applied in motor tasks more heavily dependent on proprioceptive feedback was not clear. To address this, we have tested groups of younger and older adults on a force-field adaptation task under either full or limited visual feedback conditions and examined how performance was related to dynamic proprioceptive acuity. Adaptive performance was similar between the age groups, regardless of visual feedback condition, although older adults showed increased after-effects. Physically inactive individuals made larger systematic (but not variable) proprioceptive errors, irrespective of age. However, dynamic proprioceptive acuity was unrelated to adaptation and there was no consistent evidence of proprioceptive recalibration with adaptation to the force-field for any group. Finally, in spite of clear age-dependent loss of spatial working memory capacity, we found no relationship between memory capacity and adaptive performance or proprioceptive acuity. Thus, non-clinical levels of deficit in dynamic proprioception, due to age or physical inactivity, do not affect force-field adaptation, even under conditions of limited visual feedback that might require greater proprioceptive control. Supplementary Information The online version contains supplementary material available at 10.1007/s00221-020-05997-3.

Desmoplastic small round cell tumor (DSRCT) is a rare and aggressive soft-tissue malignancy with a poor overall survival and no effective therapeutic options. The tumor is believed to be dependent on the continued activity of the oncogenic EWS-WT1 transcription factor. However, the dependence of the tumor on EWS-WT1 has not been well established. In addition, there are no studies exploring the downstream transcriptional program across multiple cell lines. In this study, we have developed a novel approach to selectively silence EWS-WT1 without impacting either wild-type EWSR1 or WT1. We show a clear dependence of the tumor on EWS-WT1 in two different cell lines, BER and JN-DSCRT-1. In addition, we identify and validate important downstream target pathways commonly dysregulated in other translocation-positive sarcomas, including PRC2, mTOR, and TGFB. Surprisingly, there is striking overlap between the EWS-WT1 and EWS-FLI1 gene signatures, despite the fact that the DNA-binding domain of the fusion proteins, WT1 and FLI1, is structurally unique and classified as different types of transcription factors. This study provides important insight into the biology of this disease relative to other translocation-positive sarcomas, and the basis for the therapeutic targeting of EWS-WT1 for this disease that has limited therapeutic options.

During normal healthy ageing there is a decline in the ability to control simple movements, characterised by increased reaction times, movement durations and variability. There is also growing evidence of age-related proprioceptive loss which may contribute to these impairments. However, this relationship has not been studied in detail for the upper limb. We recruited 20 younger adults (YAs) and 31 older adults (OAs) who each performed 2 tasks on a 2D robotic manipulandum. The first assessed dynamic proprioceptive acuity using active, multi-joint movements constrained by the robot to a pre-defined path. Participants made perceptual judgements of the lateral position of the unseen arm. The second task required fast, accurate and discrete movements to the same targets in the absence of visual feedback of the hand, and without robotic intervention. We predicted that the variable proprioceptive error (uncertainty range) assessed in Task 1 would be increased in physically inactive OAs and would predict increased movement variability in Task 2. Instead we found that physically inactive OAs had larger systematic proprioceptive errors (bias) than YAs (t[33] = 2.8, p = 0.009), and neither proprioceptive uncertainty nor bias was related to motor performance in either age group (all regression model R2 ≤ 0.06). We suggest that previously reported estimates of proprioceptive decline with ageing may be exaggerated by task demands and that the extent of these deficits is unrelated to control of discrete, rapid movement. The relationship between dynamic proprioceptive acuity and movement control in other tasks with greater emphasis on online feedback is still unclear and warrants further investigation.

The establishment of latent HIV-1 reservoirs in terminally differentiated cells represents a major impediment to the success of antiretroviral therapies. Notably, macrophages (Mϕs) are susceptible to HIV-1 infection and recent evidence suggests that they may be involved in long-term HIV-1 persistence. While the extensive functional heterogeneity seen across the Mϕ cell lineage parallels the spectrum of HIV-1 susceptibility reported across these cell subsets, the facets of Mϕ HIV-1 resistance and susceptibility remain to be fully defined. Notably, the differentiation of most Mϕ subsets depends on signaling through the macrophage colony-stimulating factor receptor (M-CSFR), which in addition to M-CSF, is now known to bind the unrelated interleukin-34 (IL-34) cytokine. The biological need for two M-CSFR ligands awaits full elucidation. Here, we report that Mϕs differentiated from human peripheral blood monocytes with IL-34 are substantially more resistant to HIV-1 infection than M-CSF-derived Mϕs. Moreover, while both Mϕ subsets express comparable surface protein levels of the HIV-1 receptor and co-receptor, CD4 and CCR5 respectively, the IL-34-Mϕs express significantly greater levels of pertinent restriction factor genes, potentially accounting for their greater resistance to HIV-1 infection than that observed in M-CSF-Mϕs. Together, our findings underline previously unexplored differentiation pathways resulting in HIV-1-susceptible and resistant Mϕ subsets and pave the way for further research that may overcome one of the last major hurdles in developing more successful antiretroviral therapy.

Recent global epidemics of viral infection such as Zika virus (ZIKV) and associated birth defects from maternal-fetal viral transmission highlights the critical unmet need for experimental models that adequately recapitulates the biology of the human maternal-fetal interface and downstream fetal development. Herein, we report an in vitro biomimetic placenta-fetus model of the maternal-fetal interface and downstream fetal cells. Using a tissue engineering approach, we built a 3D model incorporating placental trophoblast and endothelial cells into an extracellular matrix environment and validated formation of the maternal-fetal interface. We utilized this model to study ZIKV exposure to the placenta and neural progenitor cells. Our results indicated ZIKV infects both trophoblast and endothelial cells, leading to a higher viral load exposed to fetal cells downstream of the barrier. Viral inhibition by chloroquine reduced the amount of virus both in the placenta and transmitted to fetal cells. A sustained downstream neural cell viability in contrast to significantly reduced viability in an acellular model indicates that the placenta sequesters ZIKV consistent with clinical observations. These findings suggest that the placenta can modulate ZIKV exposure-induced fetal damage. Moreover, such tissue models can enable rigorous assessment of potential therapeutics for maternal-fetal medicine.

Purpose: The goal of this study was to identify second-generation mithramycin analogues that better target the EWS-FLI1 transcription factor for Ewing sarcoma. We previously established mithramycin as an EWS-FLI1 inhibitor, but the compound's toxicity prevented its use at effective concentrations in patients. Experimental Design: We screened a panel of mithralogs to establish their ability to inhibit EWS-FLI1 in Ewing sarcoma. We compared the IC50 with the MTD established in mice to determine the relationship between efficacy and toxicity. We confirmed the suppression of EWS-FLI1 at the promoter, mRNA, gene signature, and protein levels. We established an improved therapeutic window by using time-lapse microscopy to model the effects on cellular proliferation in Ewing sarcoma cells relative to HepG2 control cells. Finally, we established an improved therapeutic window using a xenograft model of Ewing sarcoma. Results: EC-8105 was found to be the most potent analogue and was able to suppress EWS-FLI1 activity at concentrations nontoxic to other cell types. EC-8042 was substantially less toxic than mithramycin in multiple species but maintained suppression of EWS-FLI1 at similar concentrations. Both compounds markedly suppressed Ewing sarcoma xenograft growth and inhibited EWS-FLI1 in vivo. Conclusions: These results provide a basis for the continued development of EC-8042 and EC-8105 as EWS-FLI1 inhibitors for the clinic. Clin Cancer Res; 22(16); 4105–18. ©2016 AACR.

The text of this abstract has not been printed at the request of the authors due to intellectual property matters. Citation Format: Maggie H. Chasse, Benjamin K. Johnson, Elissa A. Boguslawski, Katie M. Sorensen, Ian Beddows, Zachary Madaj, Gabrielle E. Foxa, Kristin J. Rybski, Susan M. Kitchen-Goosen, Bart O. Williams, Timothy J. Triche Jr., Patrick J. Grohar. Mithramycin evicts SWI/SNF from chromatin to induce epigenetic reprogramming in rhabdoid tumor [abstract]. In: Proceedings of the AACR Special Conference on the Advances in Pediatric Cancer Research; 2019 Sep 17-20; Montreal, QC, Canada. Philadelphia (PA): AACR; Cancer Res 2020;80(14 Suppl):Abstract nr A75.

Soil-transmitted nematodes infect over a billion people and place several billion more at risk of infection. Hookworm disease is the most significant of these soil-transmitted nematodes, with over 500 million people infected. Hookworm infection can result in debilitating and sometimes fatal iron-deficiency anemia, which is particularly devastating in children and pregnant women. Currently, hookworms and other soil-transmitted nematodes are controlled by administration of a single dose of a benzimidazole to targeted populations in endemic areas. While effective, people are quickly re-infected, necessitating frequent treatment. Widespread exposure to anthelmintic drugs can place significant selective pressure on parasitic nematodes to generate resistance, which has severely compromised benzimidazole anthelmintics for control of livestock nematodes in many areas of the world. Here we report, to our knowledge, the first naturally occurring multidrug-resistant strain of the canine hookworm Ancylostoma caninum. We reveal that this isolate is resistant to fenbendazole at the clinical dosage of 50 mg/kg for 3 days. Our data shows that this strain harbors a fixed, single base pair mutation at amino acid 167 of the β-tubulin isotype 1 gene, and by using CRISPR/Cas9 we demonstrate that introduction of this mutation into the corresponding amino acid in the orthologous β-tubulin gene of Caenorhabditis elegans confers a similar level of resistance to thiabendazole. We also show that the isolate is resistant to the macrocyclic lactone anthelmintic ivermectin. Understanding the mechanism of anthelmintic resistance is important for rational design of control strategies to maintain the usefulness of current drugs, and to monitor the emergence of resistance. The isolate we describe represents the first multidrug-resistant strain of A. caninum reported, and our data reveal a resistance marker that can emerge naturally in response to heavy anthelminthic treatment.

International audience; It is uncertain how vision and proprioception contribute to adaptation of voluntary arm movements. In normal participants, adaptation to imposed forces is possible with or without vision, suggesting that proprioception is sufficient; in participants with proprioceptive loss (PL), adaptation is possible with visual feedback, suggesting that proprioception is unnecessary. In experiment 1 adaptation to, and retention of, perturbing forces were evaluated in three chronically deafferented participants. They made rapid reaching movements to move a cursor toward a visual target, and a planar robot arm applied orthogonal velocity-dependent forces. Trial-by-trial error correction was observed in all participants. Such adaptation has been characterized with a dual-rate model: a fast process that learns quickly, but retains poorly and a slow process that learns slowly and retains well. Experiment 2 showed that the PL participants had large individual differences in learning and retention rates compared to normal controls. Experiment 3 tested participants' perception of applied forces. With visual feedback, the PL participants could report the perturbation's direction as well as controls; without visual feedback, thresholds were elevated. Experiment 4 showed, in healthy participants, that force direction could be estimated from head motion, at levels close to the no-vision threshold for the PL participants. Our results show that proprioceptive loss influences perception, motor control and adaptation but that proprioception from the moving limb is not essential for adaptation to, or detection of, force fields. The differences in learning and retention seen between the three deafferented participants suggest that they achieve these tasks in idiosyncratic ways after proprioceptive loss, possibly integrating visual and vestibular information with individual cognitive strategies.

Fecal glucocorticoid metabolites are increasingly used to investigate physiological stress. However, it is crucial for researchers to simultaneously investigate the effects of reproductive state because estradiol and placental hormones can affect circulating glucocorticoid concentrations. Reports on the relationships between glucocorticoids and reproductive state are inconsistent among females. Unlike several primate species that have heightened glucocorticoid activity during lactation, humans experience reduced glucocorticoid activity during lactation. Rather than a taxonomic difference, we hypothesize that this is a result of different environmental stressors, particularly the threat of infanticide. Here, we expand the number of wild primate species tested by validating a glucocorticoid assay for female Geoffroy's spider monkeys. We investigate the effects of reproductive state on their glucocorticoid concentrations. Utilizing a routine veterinary exam on a captive population, we determined that fecal glucocorticoid metabolites increase in response to a stressor (anesthesia), and this rise is detected approximately 24 hr later. Additionally, we found that extracted hormone patterns in a wild population reflected basic reproductive biology-estradiol concentrations were higher in cycling than lactating females, and in lactating females with older offspring who were presumably resuming their cycle. However, we found that estradiol and glucocorticoid concentrations were significantly correlated in lactating but not cycling females. Similarly, we found that reproductive state and estradiol concentration, but not stage of lactation, predicted glucocorticoid concentrations. Unlike patterns in several other primate species that face a relatively strong threat of infanticide, lactating spider monkeys experience reduced glucocorticoid activity, possibly due to attenuating effects of oxytocin and lower male-initiated aggression than directed at cycling females. More broadly, we conclude that future studies using fecal glucocorticoid metabolites to index stress should consider that reproductive state might confound glucocorticoid measurements. Am. J. Primatol. 77:925-935, 2015. © 2015 Wiley Periodicals, Inc.

Background: The outbreak of Zika virus (ZIKV) infection in Brazil has raised concerns that infection during pregnancy could cause microcephaly and other severe neurodevelopmental malformations in the fetus. The mechanisms by which ZIKV causes fetal abnormalities are largely unknown. The importance of pre-infection with dengue virus (DENV), or other flaviviruses endemic to Brazil, remains to be investigated. It has been reported that antibodies directed against DENV can increase ZIKV infectivity by antibody dependent enhancement (ADE), suggesting that a history of prior DENV infection might worsen the outcome of ZIKV infection.Methods: We used bioinformatics tools to design 18 peptides from the ZIKV envelope containing predicted HLA-I T-cell epitopes and investigated T-cell cross-reactivity between ZIKV-infected individuals and DENV-vaccinated subjects by IFNg ELISPOT.Results: Three peptides induced IFNg production in both ZIKV-infected subjects and in DENV-vaccinated individuals. Flow cytometry indicated that 1 ZIKV peptide induced a CD4+ T-cell response in DENV-vaccinated subjects.Conclusions: We demonstrated that vaccination against DENV induced a T-cell response against ZIKV and identified one such CD4+ T-cell epitope. The ZIKV-reactive CD4+ T cells induced by DENV vaccination and identified in this study could contribute to the appearance of cross-reactive antibodies mediating ADE.

BACKGROUND: Ewing sarcoma (ES) is the second most common pediatric bone cancer. ES cells are dependent on the activity of the EWS/FLI1 transcription factor for cell survival. EWS/FLI1 drives proliferation, dysregulates the cell cycle and establishes a block in differentiation. Despite the known dependence of Ewing sarcoma on EWS/FLI1, the exact identity of all the downstream targets of EWS/FLI1 is not known. While gene signatures have been reported for EWS/FLI1, these fail to capture the marked heterogeneity in the EWS/FLI1 transcriptome or correlate these expression changes with cellular phenotype. METHODS: Next generation RNA sequencing with deep coverage was used to generate a comprehensive analysis of the EWS/FLI1 transcriptome by silencing EWS/FLI1 in 5 different ES cell lines at two different time points. These results were organized into gene expression networks and signaling nodes were identified by weighted correlation network analysis (WGCNA). The networks were validated by a 500-gene signature of EWS/FLI1 targets using a next generation sequencing capture assay and the Illumina platform. The cellular endpoint of EWS/FLI1 silencing was evaluated by caspase 3,7 cleavage to measure apoptosis, BGAL staining to measure senescence and scratch assays to capture migration. RESULTS: Next generation RNA sequencing with siRNA silencing of EWS/FLI1 revealed marked heterogeneity in the EWS/FLI1 transcriptome among cell lines. Targets of EWS/FLI1 that were common across cell lines exhibited small magnitude gene expression changes (log fold 2). When the log fold change in expression was restricted to greater than 2, only 13 common repressed targets and 4 common induced targets were identified in all 5 cell lines. In addition, silencing of EWS/FLI1 induced dramatically different phenotypes across cell lines, where p53 wild-type cell lines underwent senescence and the p53 mutant cell lines did not. Most cell lines did not continue to proliferate with EWS/FLI1 silencing with the exception being the widely used A673 cell line. Cellular migration did not change with EWS/FLI1 silencing in many of the cell lines in contrast to A673 cells. CONCLUSIONS: EWS/FLI1 drives different gene expression signatures and different phenotypes in unique cell lines. This diversity needs to be considered as EWS/FLI1 directed therapies are developed. Citation Format: Susan M. Kitchen-Goosen, Megan J. Bowman, Marie Adams, Penny Berger, Patrick J. Grohar. The EWS/FLI1 transcriptome is characterized by marked heterogeneity across Ewing sarcoma cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3356.

Behavioral flexibility is considered by some to be one of the hallmarks of advanced cognitive ability. One measure of behavioral flexibility is how subjects respond to novel objects. Despite growing interest in comparative cognition, no comparative research on neophilia in wild primates has been conducted. Here, we compare responses to novel objects in wild chacma baboons (Papio ursinus) and geladas (Theropithecus gelada). Baboons and geladas are closely related taxa, yet they differ in their ecology and degree of social tolerance: (1) baboons are habitat and dietary generalists, whereas geladas have one of the most specialized primate diets (90% grass); (2) baboons exhibit an aversion toward extra-group individuals, whereas geladas typically exhibit an attraction toward them. Using subjects of all age and sex classes, we examined responses to three different objects: a plastic doll, a rubber ball, and a metal can. Overall, baboon subjects exhibited stronger responses to the objects (greater neophilia and exploration) than gelada subjects, yet we found no evidence that the geladas were afraid of the objects. Furthermore, baboons interacted with the objects in the same way they might interact with a potential food item. Responses were unrelated to sex, but immatures showed more object exploration than adults. Results corroborate novel object research conducted in captive populations and suggest that baboons and geladas have differences in behavioral flexibility (at least in this cognitive domain) that have been shaped by ecological (rather than social) differences between the two species.

There is a great need to develop novel approaches to target oncogenic transcription factors with small molecules. Ewing sarcoma is emblematic of this need, as it depends on the continued activity of the EWS-FLI1 transcription factor to maintain the malignant phenotype. We have previously shown that the small molecule trabectedin interferes with EWS-FLI1. Here, we report important mechanistic advances and a second-generation inhibitor to provide insight into the therapeutic targeting of EWS-FLI1. We discovered that trabectedin functionally inactivated EWS-FLI1 by redistributing the protein within the nucleus to the nucleolus. This effect was rooted in the wild-type functions of the EWSR1, compromising the N-terminal half of the chimeric oncoprotein, which is known to be similarly redistributed within the nucleus in the presence of UV light damage. A second-generation trabectedin analogue lurbinectedin (PM01183) caused the same nuclear redistribution of EWS-FLI1, leading to a loss of activity at the promoter, mRNA, and protein levels of expression. Tumor xenograft studies confirmed this effect, and it was increased in combination with irinotecan, leading to tumor regression and replacement of Ewing sarcoma cells with benign fat cells. The net result of combined lurbinectedin and irinotecan treatment was a complete reversal of EWS-FLI1 activity and elimination of established tumors in 30% to 70% of mice after only 11 days of therapy. Our results illustrate the preclinical safety and efficacy of a disease-specific therapy targeting the central oncogenic driver in Ewing sarcoma. Cancer Res; 76(22); 6657–68. ©2016 AACR.

Abstract Group members do not always act cohesively when facing extra-group rivals. When benefits such as group-defence are not monopolizable, it poses an economics problem: who should contribute to public goods and who should freeload? A collective action framework compliments existing theoretical explanations for cooperation, and provides testable hypotheses about group-level behaviour based on individual costs and benefits. Using this approach, we review research on intergroup encounters in non-human primates published over the last 20 years, focusing on participation by different classes of individuals. While food- and mate-defence explain much between-sex variation in participation, rank and reproductive access frequently explain within-sex variation. In some species, individuals may use intergroup interactions to survey potential transfer locations and mating options, which might coincidently intimidate rivals. Experimental evidence suggests that when intergroup dominance is based on relative number of fighters, individual participation still varies with sex, rank, companion behaviour and dependent offspring presence. Relatively few studies have examined how factors such as relationships within and between groups or individual temperament mediate aggression. Long-term studies of multiple habituated groups and methodological advances (e.g., playback experiments) will continue to improve our understanding of how complex group-level patterns are predictable when viewed from an individual perspective.

Cross-fostering has been attempted opportunistically with endangered canids as a means of increasing populations. Due to the usefulness of cross-fostering for conservation, an understanding of factors influencing success rates is essential. Using captive coyotes (Canis latrans) as a model, we assessed the willingness of adult pairs to foster young born to other parents. We assessed the efficacy of fostering pups into existing litters (augmentation) and completely switching litters (replacement). We augmented four litters with two pups of similar age when pups were

We have previously shown that activation of P2X purinoceptors in the subpostremal nucleus tractus solitarius (NTS) produces a rapid bradycardia and hypotension. This bradycardia could occur via sympathetic withdrawal, parasympathetic activation, or a combination of both mechanisms. Thus we investigated the relative roles of parasympathetic activation and sympathetic withdrawal in mediating this bradycardia in chloralose-urethane anesthetized male Sprague-Dawley rats. Microinjections of the selective P2X purinoceptor agonist α,β-methylene ATP (25 pmol/50 nl and 100 pmol/50 nl) were made into the subpostremal NTS in control animals, after atenolol (2 mg/kg iv), a β1-selective antagonist, and after atropine methyl bromide (2 mg/kg iv), a muscarinic receptor antagonist. The bradycardia observed with activation of P2X receptors at the low dose of the agonist is mediated almost entirely by sympathetic withdrawal. After β1-adrenergic blockade, the bradycardia was reduced to just −5.1 ± 0.5 versus −28.8 ± 5.1 beats/min in intact animals. Muscarinic blockade did not produce any significant change in the bradycardic response at the low dose. At the high dose, both β1-adrenergic blockade and muscarinic blockade attenuated the bradycardia similarly, −37.4 ± 6.4 and −40.6 ± 3.7 beats/min, respectively, compared with −88.0 ± 11 beats/min in control animals. Double blockade of both β1-adrenergic and muscarinic receptors virtually abolished the response (−2.5 ± 0.8 beats/min). We conclude that the relative contributions of parasympathetic activation and sympathetic withdrawal are dependent on the extent of P2X receptor activation.

Swift foxes (Vulpes velox) and coyotes (Canis latrans) are sympatric canids distributed throughout many regions of the Great Plains of North America. The prevalence of canid diseases among these two species where they occur sympatrically is presently unknown. From January 1997 to January 2001, we collected blood samples from 89 swift foxes and 122 coyotes on the US Army Piñon Canyon Maneuver Site, Las Animas County, SE Colorado (USA). Seroprevalence of antibodies against canine parvovirus (CPV) was 71% for adult (9 mo old) and 38% for juvenile (or = 9 mo old) swift foxes. Adult (1 yr old) and juvenile (1 yr old) coyotes had a seroprevalence for CPV of 96% and 78%, respectively. Presence of antibodies against canine distemper virus (CDV) was 5% for adult foxes and 0% for juvenile foxes. Seroprevalence of CDV was 46% for adult coyotes and 18% for juvenile coyotes. No swift foxes had canine adenovirus (CAV) antibodies, whereas 81% and 63% of adult and juvenile coyotes, respectively, had antibodies for CAV. Seroprevalence of antibodies against Yersinia pestis was 68% among adult foxes and 34% among juvenile swift foxes. Seroprevalence of Y. pestis antibodies was 90% and 70% for adult and juvenile coyotes, respectively. No swift foxes had antibodies against Francisella tularensis, whereas seroprevalence was 4% among both adult and juvenile coyotes. Antibodies against CPV and plague were common in both species, whereas antibodies against CDV and CAV were more prevalent in coyotes compared to swift foxes.

Transitional cell carcinoma (TCC) of the bladder is the most common urothelial malignancy. Sites of metastatic spread are usually predictable; typically the pelvic and para-aortic lymphatics, followed by extra-vesical tissues, i.e. liver, lung, bone and adrenal glands. There is currently little literature describing metastatic spread to the head and neck region, and none to the authors’ knowledge, of spread to the parotid salivary glands. This report describes a rare case of parotid metastasis of bladder TCC in a 75-year-old Caucasian woman.

SUMMARY 1. Studies are described that indicate that stimulation of different purinergic receptor subtypes (A1, A2A and P2X) located in the sub-postremal nucleus tractus solitarius (NTS) evokes qualitatively and quantitatively different regional haemodynamic and efferent sympathetic responses. 2. Stimulation of A2A receptors evoked the most diverse pattern of regional sympathetic responses: preganglionic adrenal nerve activity (pre-ASNA) was increased, lumbar sympathetic nerve activity (LSNA) did not change, while renal (RSNA) and post-ganglionic adrenal (post-ASNA) sympathetic nerve activity was decreased. Stimulation of A1 receptors evoked qualitatively uniform, although quantitatively different, sympathoactivation: pre-ASNA > RSNA >LSNA. Stimulation of P2X receptors evoked qualitatively uniform, although quantitatively different, sympathoinhibition: RSNA = post-ASNA > LSNA = pre-ASNA. 3. These qualitatively and quantitatively different patterns of regional sympathetic responses strongly suggest that purinergic receptor subtypes may be specifically located and differentially expressed on NTS neurons/neural terminals that control different sympathetic outputs. Different NTS purinoceptors may contribute to patterned autonomic responses observed in specific physiological or pathological situations.

Previously, we have shown that activation of adenosine A2a receptors in the subpostremal nucleus tractus solitarii (NTS) via microinjection of the selective A2a receptor agonist CGS-21680 elicits potent, dose-dependent decreases in mean arterial pressure and preferential, marked hindlimb vasodilation. Although A2a receptor activation does not change lumbar sympathetic nerve activity, it does markedly enhance the preganglionic adrenal sympathetic nerve activity, which will increase epinephrine release and could subsequently elicit hindlimb vasodilation via activation of β2-adrenergic receptors. Therefore we investigated whether this hindlimb vasodilation was due to neural or humoral mechanisms. In chloralose-urethan-anesthetized male Sprague-Dawley rats, we monitored cardiovascular responses to stimulation of NTS adenosine A2a receptors (CGS-21680, 20 pmol/50 nl) in the intact control animals; after pretreatment with propranolol (2 mg/kg iv), a β-adrenergic antagonist; after bilateral lumbar sympathectomy; after bilateral adrenalectomy; and after combined bilateral lumbar sympathectomy and adrenalectomy. After β-adrenergic blockade, stimulation of NTS adenosine A2a receptors produced a pressor response and a hindlimb vasoconstriction. Lumbar sympathectomy reduced the vasodilation seen in the intact animals by ∼40%, and adrenalectomy reduced it by ∼80%. The combined sympathectomy and adrenalectomy virtually abolished the hindlimb vasodilation evoked by NTS A2a receptor activation. We conclude that the preferential, marked hindlimb vasodilation produced by stimulation of NTS adenosine A2a receptors is mediated by both the efferent sympathetic nerves directed to the hindlimb and the adrenal glands via primarily a β-adrenergic mechanism.

Activity patterns in animals are influenced by a number of factors, including the animal's physiological adaptations, prey availability and distribution, and disturbances caused by predators and humans. We compared coyote (Canis latrans) activity patterns estimated using radio-tracking locations between 1983 and 1988 with those documented between 1996 and 1997 on the Pinon Canyon Maneuver Site, in southeastern Colorado. We tested the hypothesis that changes in the type of disturbance experienced by coyotes would result in changes in their activity patterns. Disturbance experienced by the coyote population studied during 1983-1988, included >50 years of intense exploitation (shooting and trapping by ranchers) and intensive removal efforts using aerial gunning. In contrast, coyotes tracked during 1996-1997 experienced some periodic disturbance from army maneuvers occurring in the area, but were not exposed to any direct form of persecution (e.g., shooting). From August 1983 to July 1988, 49 coyotes (26 males and 23 females) were tracked for >2400 h using radiotelemetry. From April 1996 to August 1997, 22 coyotes (12 males and 10 females) were tracked for >950 h. The average rate of diurnal movement of the coyotes in the 1996-1997 field study (mean = 0.97 km/h) was significantly higher than that of the coyotes in the 1983-1988 field study (mean = 0.68 km/h). This occurred despite no significant increase in the overall (24 h) rate of movement between the two field studies. Estimates of prey use by the coyotes in both field studies were obtained, to test an alternate hypothesis that prey switching might explain the changes in coyote movement patterns. However, there was no significant difference between the frequency of occurrence of diurnally versus nocturnally active mammalian prey species in the diets of coyotes in any season or overall between the 1983-1988 and 1996-1997 field studies. This study demonstrated that coyote activity patterns can be influenced by the type of disturbance experienced by the animal. A coyote population that had historically been exposed to human persecution shifted to higher levels of diurnal activity when exploitation ceased.

European guidelines recommend the routine offer of an HIV test in patients with a number of AIDS-defining and non-AIDS conditions believed to share an association with HIV; so called indicator conditions (IC). Adherence with this guidance across Europe is not known. We audited HIV testing behaviour in patients accessing care for a number of ICs. Participating centres reviewed the case notes of either 100 patients or of all consecutive patients in one year, presenting for each of the following ICs: tuberculosis, non-Hodgkins lymphoma, anal and cervical cancer, hepatitis B and C and oesophageal candidiasis. Observed HIVpositive rates were applied by region and IC to estimate the number of HIV diagnoses potentially missed. Outcomes examined were: HIV test rate (% of total patients with IC), HIV test accepted (% of tests performed/% of tests offered) and new HIV diagnosis rate (%). There were 49 audits from 23 centres, representing 7037 patients. The median test rate across audits was 72% (IQR 32-97), lowest in Northern Europe (median 44%, IQR 22-68%) and highest in Eastern Europe (median 99%, IQR 86-100). Uptake of testing was close to 100% in all regions. The median HIV+ rate was 0.9% (IQR 0.0-4.9), with 29 audits (60.4%) having an HIV+ rate >0.1%. After adjustment, there were no differences between regions of Europe in the proportion with >0.1% testing positive (global p = 0.14). A total of 113 patients tested HIV+. Applying the observed rates of testing HIV+ within individual ICs and regions to all persons presenting with an IC suggested that 105 diagnoses were potentially missed. Testing rates in well-established HIV ICs remained low across Europe, despite high prevalence rates, reflecting missed opportunities for earlier HIV diagnosis and care. Significant numbers may have had an opportunity for HIV diagnosis if all persons included in IC audits had been tested., SCOPUS: ar.j, info:eu-repo/semantics/published

Rho GTPase-effector mammalian diaphanous (mDia)–related formins assemble nonbranched actin filaments as part of cellular processes, including cell division, filopodia assembly, and intracellular trafficking. Whereas recent efforts have led to thorough characterization of formins in cytoskeletal remodeling and actin assembly in vitro, little is known about the role of mDia proteins in vivo. To fill this knowledge gap, the Drf1 gene, which encodes the canonical formin mDia1, was targeted by homologous recombination. Upon birth, Drf1+/− and Drf1−/− mice were developmentally and morphologically indistinguishable from their wild-type littermates. However, both Drf1+/− and Drf1−/− developed age-dependent myeloproliferative defects. The phenotype included splenomegaly, fibrotic and hypercellular bone marrow, extramedullary hematopoiesis in both spleen and liver, and the presence of immature myeloid progenitor cells with high nucleus-to-cytoplasm ratios. Analysis of cell surface markers showed an age-dependent increase in the percentage of CD11b+-activated and CD14+-activated monocytes/macrophages in both spleen and bone marrow in Drf1+/− and Drf1−/− animals. Analysis of the erythroid compartment showed a significant increase in the proportion of splenic cells in S phase and an expansion of erythroid precursors (TER-119+ and CD71+) in Drf1-targeted mice. Overall, knocking out mDia1 expression in mice leads to a phenotype similar to human myeloproliferative syndrome (MPS) and myelodysplastic syndromes (MDS). These observations suggest that defective DRF1 expression or mDia1 function may contribute to myeloid malignancies and point to mDia1 as an attractive therapeutic target in MDS and MPS. [Cancer Res 2007;67(16):7565–71]

Activated T cells rapidly assemble filamentous (F-) actin networks in response to ligation of the T cell receptor or upon interaction with adhesive stimuli in order to facilitate cell migration and the formation of the immune synapse. Branched filament assembly is crucial for this process and is dependent upon activation of the Arp2/3 complex by the actin nucleation-promoting factor Wiskott-Aldrich Syndrome protein (WASp). Genetic disruption of the WAS gene has been linked to hematopoietic malignancies and various cytopenias. Although the contributions of WASp and Arp2/3 to T cell responses are fairly well characterized, the role of the mammalian Diaphanous (mDia)-related formins, which both nucleate and processively elongate non-branched F-actin, has not been demonstrated. Here, we report the effects on T cell development and function following the knock out of the murine Drf1 gene encoding the canonical formin p140mDia1. Drf1(-/-) mice develop lymphopenia characterized by diminished T cell populations in lymphoid tissues. Consistent with a role for p140mDia1 in the regulation of the actin cytoskeleton, isolated Drf1(-/-) splenic T cells adhered poorly to extracellular matrix proteins and migration in response to chemotactic stimuli was completely abrogated. Both integrin and chemokine receptor expression was unaffected by Drf1(-/-) targeting. In response to proliferative stimuli, both thymic and splenic Drf1(-/-) T cells failed to proliferate; ERK1/2 activation was also diminished in activated Drf1(-/-) T cells. These data suggest a central role for p140mDia1 in vivo in dynamic cytoskeletal remodeling events driving normal T cell responses.

While the murine equivalent to “5q– syndrome” should map to the distal arms of murine chromosomes 11 and 18, molecular attempts have been unsuccessful in generating a reliable mouse model of myelodysplasia leading to exacerbated myeloid leukemia. Genetic studies suggest interactions between individual proteins in this locus suppress tumor initiation. More specifically, cellular and biochemical studies demonstrated collaboration between two proteins in the commonly deleted regions of 5q– syndrome: mDia1 (mammalian Diaphanous-related Rho GTPase effector formin) and APC (adenomatous polypolis coli). Therefore, we generated mice compounding these two putative disease alleles. Because both mDia1 knockout [Drf1(–/–)] mice and Apcmin (min = multiple intestinal neoplasia; haploinsufficient) mice develop age-dependent myelodysplastic syndrome (MDS), we hypothesized that a mouse strain lacking both proteins will exhibit a more severe MDS phenotype than either mutant allele alone. To define the disease caused by mutation in Drf1 and Apcmin, we monitored erythropoiesis, myelopoiesis, and overall survival. At all ages, mice with both mutant alleles exhibit granulocytosis. Anemia occurs in aged mice (>270 days), exhibiting drastic differences in blood cell morphology that corresponded to differences in long-term survival in comparison to either Drf1–/– or Apcmin animals. Furthermore, immunophenotyping of all subpopulations indicate an accumulation of CD71+/TER-119+ cells in mice with both mutant alleles, however no significant differences in CD11b expression appeared among CD45+ bone marrow subpopulations. Our studies indicate that combined loss of mDia1 and APC expression and function direct disease progression. We are currently defining the hematopoietic precursor cell responsible for anemia and bone marrow failure. This enhanced mouse model also serves as molecular laboratory to study disease progression and to test potential therapeutics under development in our laboratory. Citation Format: Julie Davis Turner, Susan Kitchen-Goosen, Andrew M. Howard, Heather Schumacher, Art S. Alberts. Modeling MDS in mice through precise molecular lesions in APC and mDia1. [abstract]. In: Proceedings of the AACR Special Conference on Hematologic Malignancies: Translating Discoveries to Novel Therapies; Sep 20-23, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(17 Suppl):Abstract nr B42.

We have previously shown that P2x purinoceptor activation in the subpostremal nucleus tractus solitarius (NTS) produces dose-dependent decreases in mean arterial pressure (MAP), heart rate, efferent sympathetic nerve activity, and significant peripheral vasodilation. However, the relative roles of cardiac output (CO) and total peripheral resistance (TPR) in mediating this depressor response are unknown. Bradycardia does not necessarily result in decreased CO, because, with the greater filling time, stroke volume may increase such that CO may be unchanged. We measured changes in CO (via a chronically implanted flow probe on the ascending aorta) and MAP in α-chloralose- and urethane-anesthetized male Sprague-Dawley rats in response to microinjection of the selective P2x purinoceptor agonist α,β-methylene ATP (25 and 100 pmol/50 nl) into the subpostremal NTS. TPR was calculated as MAP/CO. At the low dose of NTS P2x purinoceptor agonist, the reduction in MAP was primarily mediated by reductions in TPR (−31.3 ± 3.3%), not CO (−8.7 ± 1.7%). At the high dose, both CO (−34.4 ± 6.6%) and TPR (−40.2 ± 2.5%) contribute to the reduction in MAP. We conclude that the relative contribution of CO and TPR to the reduction in MAP evoked by NTS P2x purinoceptor activation is dependent on the extent of P2x purinoceptor activation.

Rho small GTP-binding proteins locally manage Diaphanous-related formins as they construct the cytoskeletal infrastructure that supports essential changes in cell morphology. Rho binding unlatches formin intramolecular autoinhibition. The autoinhibition is mediated by the Dia-inhibitory and -autoregulatory domains (DID & DAD) which flank the formin homology-2 (FH2) domain. The free FH2 domain guides the assembly of the linear actin and microtubule structures required for adhesion, migration, division, and polarity establishment. Microtubule stabilization is already known to be an effective anti-cancer strategy. Combined with the knowledge that formins can act as haploinsufficient tumor suppressor genes, a screen was conducted for agents that could impair autoinhibition in order to activate formins for therapeutic benefit. We have focused on two of the primary “hits” for development. VARI01 and VARI02 are nearly identical thioamide inhibitors of DID-DAD binding (IC50 values ≤ 140 nM). Exposure of cultured tumor cells to either compound triggered the expected changes in cell structure and signaling that were consistent with formin activation. These included microtubule stabilization, F-actin assembly, the activation of the transcription factor SRF, cell-cycle arrest, and eventual apoptosis. SRF senses changes in actin dynamics to activate the expression of genes that reinforce cell structure and monitor genome integrity. VARI01 and VARI02 both exhibit anti-tumor activity in xenograft studies. Ongoing studies are aimed at identifying VARI01/VARI02-sensitive tumor types and to define other potential therapeutic applications. Significance: Microtubule stabilization by taxol has proven to be an effective anti-cancer strategy, but it has significant deleterious side effects. Small-molecule formin activation serve as a novel class of anti-cancer agents that may provide tools for unlocking the biological roles of formins in signalling and disease. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4764. doi:1538-7445.AM2012-4764

The Adenomatous polyposis coli (APC1) tumor suppressor is a multi-function protein that is involved in both the Wnt signaling cascade and cytoskeletal remodeling. Mutations or loss of APC1 can result in defects in cell migration, proliferation, and differentiation. APC frequently collaborates with the canonical Diaphanous-related formin, mDia1 (DIAPH1), which also functions to regulate cytoskeletal dynamics. APC1 and DIAPH1 are genetically linked on Chromosome 5q in a region frequently disrupted in myeloproliferative neoplasms (MPNs). Mice lacking mDia1 (Drf1) and the Apc min mice develop myelodysplastic phenotypes. The physical and functional linkage of APC and mDia1 as 5q tumor suppressors and in cytoskeletal remodeling led us to explore the role of APC in tumor suppression in myeloid-derived cell lineages. We created a mouse which lacks APC in myeloid lineage cells by breeding Apc-flox mice with mice expressing Cre recombinase under the control of lysozyme M (LyzsM) gene promoter. Lysozyme M is expressed in myeloid lineage cells including monocytes, macrophages and granulocytes. Surprisingly, the female mice develop cystic endometrial hyperplasia and are unable to sustain pregnancy. Lysozyme M expression appears to be limited to tissue resident macrophages within the uterus and was not detected in uterine epithelial cells. However, there is a small subset of uterine epithelial cells with increased β-catenin expression, suggesting that these cells lack APC. This subset of cells has elevated expression of cyclin D1, a regulator of cell cycle progression and a downstream effector of Wnt signaling, resulting in increased proliferation. Interestingly, the uterine epithelial cells with increased β-catenin expression also have increased estrogen receptor ≤ (ERα) expression. Estrogen stimulates uterine growth and unopposed estrogen signaling is a hallmark of Type I endometrial cancer. It is unclear if the endometrial hyperplasia seen in these mice is due to lysozyme M expression early on in uterine epithelial cell development or perhaps in a progenitor cell population. A second alternative is that loss of APC in tissue resident myeloid cells residing in the uterine stroma affects epithelial morphology. Regardless, these data suggest that APC plays a role in uterine tissue homeostasis, perhaps even regulating ERα expression. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3280. doi:1538-7445.AM2012-3280

The Adenomatous polyposis coli (APC1) tumor suppressor is a multi-function protein with a central role in Wnt-regulated cell growth and differentiation. Additionally, APC functions to modulate microtubule and microfilament dynamics to establish polarity in epithelia during directed cell migration and asymmetric cell division. APC frequently collaborates with the canonical Diaphanous-related formin, mDia1 (DIAPH1), which also binds microfilaments and microtubules to modulate cytoskeletal dynamics. APC1 and DIAPH1 are genetically linked on chromosome 5q in a region frequently disrupted in myeloproliferative neoplasms (MPNs). mDia1 knockout mice develop an age-dependent outgrowth of myeloid-lineage cells. The physical and functional linkage of APC and mDia1 as 5q tumor suppressors and in cytoskeletal remodeling led us to explore the role of APC in tumor suppression in myeloid-derived cell lineages affected by mDia1 knockout. To test this hypothesis in a genetic model, we created a mouse in which APC was knocked out in myeloid lineage cells by breeding the Apc-flox mice with mice expressing Cre recombinase under the control of lysozyme M (LyzsM) gene promoter. The resulting phenotype was unexpected and 100% penetrant: Female mice developed cystic endometrial hyperplasia and were unable to sustain pregnancy. Embryo implantation occurred, but embryos were resorbed mid-gestation irrespective of progeny genotype. Two hypothesized mechanisms to explain the initiation of endometrial hyperplasia are being pursued. The first hypothesis is that loss of APC impairs the homeostasis between growth and differentiation of myeloid lineage stem/progenitor cells that transdifferentiate into uterine epithelia. An alternate hypothesis is that loss of APC expression affects the inflammatory response of resident uterine macrophages that lead to defects in epithelial morphology. In either case, these genetic studies imply that APC gene defects in myeloid lineage cells may be an initiating event in endometrial cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4330. doi:10.1158/1538-7445.AM2011-4330

A 17-year-old man, a non-smoker, presented with a 4-day history of chest pain and dyspnoea. The symptoms had begun immediately after an episode of aspiration, which occurred whilst learning to be a fire eater. The patient …

The effects of johnsongrass [Sorghum halepense(L.) Pers. # SORHA] infestations were determined during a 2-yr study on sugarcane (Saccharum officinarumL. ‘CP-65-357’) yield. Both cane and sugar yields were lower (36 and 31%, respectively) in plots heavily infested with johnsongrass than in weed-free plots. Sugarcane yields were inversely influenced by johnsongrass equivalents (the sum of the values obtained by multiplying the number of clumps by their corresponding importance value for each plot). In 1983, both johnsongrass standing crop and johnsongrass equivalents correlated negatively with cane yield much better than any other factor combinations, while in 1984, the same was true for stalk population. Substantial yield reductions from johnsongrass interference were observed at johnsongrass infestation levels higher than 15 to 35%.

The c-fos gene is rapidly and transiently activated in quiescent BALB/c-3T3 cells in response to serum, platelet-derived growth factor or conditioned medium from v-sis-transformed cells. This activation occurs at the level of transcription and in the absence of new protein synthesis. Using a gel electrophoresis DNA-binding assay, we have found a DNA-binding activity in BALB/c-3T3 cells that is induced within 20 min of treatment with conditioned medium from v-sis-transformed cells. A DNA methylation interference assay has shown that this factor binds to a sequence approximately 346 base pairs upstream of the transcription initiation site of the human c-fos gene. Insulin, epidermal growth factor, and phorbol 12-myristate 13-acetate fail to induce this DNA-binding factor. Protein synthesis inhibitors do not block the induction of this activity. We propose that this factor preexists in an inactive form in quiescent cells and that its binding activity is activated in response to appropriate extracellular inducers.

Summary Cancer cells display metabolic plasticity to survive stresses in the tumor microenvironment. Cellular adaptation to energetic stress is coordinated in part by signaling through the liver kinase B1 (LKB1)-AMP-activated protein kinase (AMPK) pathway. Here, we demonstrate that miRNA-mediated silencing of LKB1 confers sensitivity of lymphoma cells to mitochondrial inhibition by biguanides. Using both classic (phenformin) and newly developed (IM156) biguanides, we demonstrate that elevated miR-17∼92 expression in Myc+ lymphoma cells promotes increased apoptosis to biguanide treatment in vitro and in vivo. This effect is driven by the miR-17-dependent silencing of LKB1, which reduces AMPK activation in response to complex I inhibition. Mechanistically, biguanide treatment induces metabolic stress in Myc+ lymphoma cells by inhibiting TCA cycle metabolism and mitochondrial respiration, exposing metabolic vulnerability. Finally, we demonstrate a direct correlation between miR-17∼92 expression and biguanide sensitivity in human cancer cells. Our results identify miR-17∼92 expression as a potential biomarker for biguanide sensitivity in malignancies.