Members of the ETS transcription factor family have been implicated in several cancers, where they are often dysregulated by genomic derangement. ETS variant 1 (ETV1) is an ETS factor gene that undergoes chromosomal translocation in prostate cancers and Ewing sarcomas, amplification in melanomas, and lineage dysregulation in gastrointestinal stromal tumors. Pharmacologic perturbation of ETV1 would be appealing in these cancers; however, oncogenic transcription factors are often deemed "undruggable" by conventional methods. Here, we used small-molecule microarray screens to identify and characterize drug-like compounds that modulate the biologic function of ETV1. We identified the 1,3,5-triazine small molecule BRD32048 as a top candidate ETV1 perturbagen. BRD32048 binds ETV1 directly, modulating both ETV1-mediated transcriptional activity and invasion of ETV1-driven cancer cells. Moreover, BRD32048 inhibits p300-dependent acetylation of ETV1, thereby promoting its degradation. These results point to a new avenue for pharmacologic ETV1 inhibition and may inform a general means to discover small molecule perturbagens of transcription factor oncoproteins.
Agricultural fungicide application has increased tenfold since 2005 in the United States. Active ingredients and formulations of strobilurin fungicides at environmentally relevant concentrations cause mortality to larval and metamorph amphibians; however, little is known about chronic exposure effects in amphibians. We exposed larval amphibians (Bufo cognatus) throughout metamorphosis to the common fungicide formulations Headline(®), Stratego(®), Quilt(®), and a control to determine effects on development and growth. Formulations were tested at 1.7, 50, and 400 ?g/L of the active strobilurin ingredient for Headline(®), Stratego(®), and Quilt(®), respectively. Fungicide exposure did not affect body mass or snout-vent length at metamorphosis. However, exposure to Headline(®) at 1.7 ?g/L increased the development rate of tadpoles by approximately 5 days compared to the control, an effect not observed for Stratego(®) and Quilt(®). Stratego(®) also caused approximately 35 % cumulative mortality. Results from the experiment suggest that chronic effects of strobilurin fungicides on development, growth, and mortality to B. cognatus are apparent at environmentally relevant concentrations.
Pollen tubes extend rapidly when pollen grains are incubated in defined media. Tube extension requires many critical functions of plant cells including molecular signaling, cytoskeleton remodeling, secretion, and cell wall synthesis. Consequently, pollen tube growth has been established as a way to conduct primary screens of chemical libraries to identify compounds that perturb key cellular processes in plants.
Lenalidomide is a drug with clinical efficacy in multiple myeloma and other B cell neoplasms, but its mechanism of action is unknown. Using quantitative proteomics, we found that lenalidomide causes selective ubiquitination and degradation of two lymphoid transcription factors, IKZF1 and IKZF3, by the CRBN-CRL4 ubiquitin ligase. IKZF1 and IKZF3 are essential transcription factors in multiple myeloma. A single amino acid substitution of IKZF3 conferred resistance to lenalidomide-induced degradation and rescued lenalidomide-induced inhibition of cell growth. Similarly, we found that lenalidomide-induced IL2 production in T cells is due to depletion of IKZF1 and IKZF3. These findings reveal a novel mechanism of action for a therapeutic agent, alteration of the activity of an E3 ubiquitin ligase leading to selective degradation of specific targets.
In an effort to improve pedestrian safety, several states in the United States changed their pedestrian laws by changing the requirement that drivers yield to pedestrians in crosswalks to a requirement that drivers stop for pedestrians in crosswalks. This study examined whether this change had an effect on pedestrian safety in the United States, with its focus on low-speed roads. To examine the association between changes in pedestrian laws and changes in pedestrian-involved fatal crashes, three approaches were employed: before-after analysis, time-series analysis, and cross-sectional analysis. Pedestrian-involved fatal traffic crashes on low-speed roads were extracted from the U.S. national fatal crash database, the Fatality Analysis Reporting System (FARS), from 1980 through 2005. This study found no statistically significant reduction in pedestrian-involved fatal crashes attributable to changes in the laws, yet this finding is not definitive because of study limitations such as the omission of relevant exposure data.
The objective of this study is to investigate cardiac bioeffects resulting from ultrasonic stimulation using a specific set of acoustical parameters. Ten Sprague-Dawley rats were anesthetized and exposed to 1-MHz ultrasound pulses of 3-MPa peak rarefactional pressure and approximately 1% duty factor. The pulse repetition frequency started slightly above the heart rate and was decreased by 1 Hz every 10 s, for a total exposure duration of 30 s. The control group was composed of five rats. Two-way analysis of variance for repeated measures and Bonferroni post hoc tests were used to compare heart rate and ejection fraction, which was used as an index of myocardial contractility. It was demonstrated for the first time that transthoracic ultrasound has the potential to decrease the heart rate by ~20%. The negative chronotropic effect lasted for at least 15 min after ultrasound exposure and there was no apparent gross damage to the cardiac tissue.
Related JoVE Video
Journal of Visualized Experiments
What is Visualize?
JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.
How does it work?
We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.
Video X seems to be unrelated to Abstract Y...
In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.