RAB27A promotes melanoma cell invasion and metastasis via regulation of pro-invasive exosomes.

Despite recent advances in targeted and immune-based therapies, advanced stage melanoma remains a clinical challenge with a poor prognosis. Understanding the genes and cellular processes that drive progression and metastasis is critical for identifying new therapeutic strategies. Here, we found that the GTPase RAB27A was overexpressed in a subset of melanomas, which correlated with poor patient survival. Loss of RAB27A expression in melanoma cell lines inhibited 3D spheroid invasion and cell motility in vitro, and spontaneous metastasis in vivo. The reduced invasion phenotype was rescued by RAB27A-replete exosomes, but not RAB27A-knockdown exosomes, indicating that RAB27A is responsible for the generation of pro-invasive exosomes. Furthermore, while RAB27A loss did not alter the number of exosomes secreted, it did change exosome size and altered the composition and abundance of exosomal proteins, some of which are known to regulate cancer cell movement. Our data suggest that RAB27A promotes the biogenesis of a distinct pro-invasive exosome population. These findings support RAB27A as a key cancer regulator, as well as a potential prognostic marker and therapeutic target in melanoma.

Enforced specificity of an animal symbiosis.

Insect diversification has been catalyzed by widespread specialization on novel hosts - a process underlying exceptional radiations of phytophagous beetles, lepidopterans, parasitoid wasps, and inordinate lineages of symbionts, predators and other trophic specialists. The strict fidelity of many such interspecies associations is posited to hinge on sensory tuning to host-derived cues, a model supported by studies of neural function in host-specific model species. Here, we investigated the sensory basis of symbiotic interactions between a myrmecophile rove beetle and its single, natural host ant species. We show that host cues trigger analogous behaviors in both ant and symbiont. Cuticular hydrocarbons - the ant's nestmate recognition pheromones - elicit partner recognition by the beetle and execution of ant grooming behavior, integrating the beetle into the colony via chemical mimicry. The beetle also follows host trail pheromones, permitting inter-colony dispersal. Remarkably, the rove beetle also performs its symbiotic behaviors with ant species separated by ~95 million years, and shows minimal preference for its natural host over non-host ants. Experimentally validated agent-based modeling supports a scenario in which specificity is enforced by physiological constraints on symbiont dispersal, and negative fitness interactions with alternative hosts, rather than via sensory tuning. Enforced specificity may be a pervasive mechanism of host range restriction of specialists embedded within host niches. Chance realization of latent compatibilities with alternative hosts may facilitate host switching, enabling deep-time persistence of obligately symbiotic lineages.

Direct and coordinate regulation of ATP-binding cassette transporter genes by Myc factors generates specific transcription signatures that significantly affect the chemoresistance phenotype of cancer cells.

Increased expression of specific ATP-binding cassette (ABC) transporters is known to mediate the efflux of chemotherapeutic agents from cancer cells. Therefore, establishing how ABC transporter genes are controlled at their transcription level may help provide insight into the role of these multifaceted transporters in the malignant phenotype. We have investigated ABC transporter gene expression in a large neuroblastoma data set of 251 tumor samples. Clustering analysis demonstrated a strong association between differential ABC gene expression patterns in tumor samples and amplification of the MYCN oncogene, suggesting a correlation with MYCN function. Using expression profiling and chromatin immunoprecipitation studies, we show that MYCN oncoprotein coordinately regulates transcription of specific ABC transporter genes, by acting as either an activator or a repressor. Finally, we extend these notions to c-MYC showing that it can also regulate the same set of ABC transporter genes in other tumor cells through similar dynamics. Overall our findings provide insight into MYC-driven molecular mechanisms that contribute to coordinate transcriptional regulation of a large set of ABC transporter genes, thus affecting global drug efflux.

Genetic factors in the pathogenesis of cholangiocarcinoma.

BACKGROUND: Cholangiocarcinoma (CC) is increasing in incidence, but its pathogenesis remains poorly understood. Chronic inflammation of the bile duct and cholestasis are major risk factors, but most cases in the West are sporadic. Genetic polymorphisms in biliary transporter proteins have been implicated in benign biliary disease and, in the case of progressive familial cholestasis, have been associated with childhood onset of CC. In the current study, five biologically plausible candidate genes were investigated: ABCB11 (BSEP), ABCB4 (MDR3), ABCC2 (MRP2), ATP8B1 (FIC1) and NR1H4 (FXR). METHODS: DNA was collected from 172 Caucasian individuals with confirmed CC. A control cohort of healthy Caucasians was formed. Seventy-three SNPs were selected using the HapMap database to capture genetic variation around the five candidate loci. Genotyping was undertaken with a competitive PCR-based system. Confirmation of Hardy-Weinberg equilibrium and Cochran-Armitage trend testing were performed using PLINK. Haplotype frequencies were compared using haplo.stats. RESULTS: All 73 SNPs were in Hardy-Weinberg equilibrium. Four SNPs in ABCB11 were associated with altered susceptibility to CC, including the V444A polymorphism, but these associations did not retain statistical significance after Bonferroni correction for multiple testing. Haplotype analysis of the genotyped SNPs in ATP8B1 identified significant differences in frequencies between cases and controls (global p value of 0.005). CONCLUSION: Haplotypes in ATP8B1 demonstrated a significant difference between CC and control groups. There was a trend towards significant association of V444A with CC. Given the biological plausibility of polymorphisms in ABCB11 and ATP8B1 as risk modifiers for CC, further study in a validation cohort is required.

Polymorphisms in Natural Killer Cell Receptor Protein 2D (NKG2D) as a Risk Factor for Cholangiocarcinoma.

BACKGROUND AND AIMS: Understanding of the significant genetic risk factors for Cholangiocarcinoma (CC) remains limited. Polymorphisms in the natural killer cell receptor G2D (NKG2D) gene have been shown to increase risk of CC transformation in patients with Primary Sclerosing Cholangitis (PSC). We present a validation study of NKG2D polymorphisms in CC patients without PSC. METHODS: Seven common Single Nucleotide Polymorphisms (SNPs) of the NKG2D gene were genotyped in 164 non-PSC related CC subjects and 257 controls with HaploView. The two SNPs that were positively identified in the previous Scandinavian study, rs11053781 and rs2617167, were included. RESULTS: The seven genotyped SNPs were not associated with risk of CC. Furthermore, haplotype analysis revealed that there was no evidence to suggest that any haplotype differs in frequency between cases and controls (P > 0.1). CONCLUSION: The common genetic variation in NKG2D does not correlate significantly with sporadic CC risk. This is in contrast to the previous positive findings in the Scandinavian study with PSC-patients. The failure to reproduce the association may reflect an important difference between the pathogenesis of sporadic CC and that of PSC-related CC. Given that genetic susceptibility is likely to be multifaceted and complex, further validation studies that include both sporadic and PSC-related CC are required.

Visual working memory capacity for objects from different categories: a face-specific maintenance effect.

The capacity of visual working memory was examined when complex objects from different categories were remembered. Previous studies have not examined how visual similarity affects object memory, though it has long been known that similar-sounding phonological information interferes with rehearsal in auditory working memory. Here, experiments required memory for two or four objects. Memory capacity was compared between remembering four objects from a single object category to remembering four objects from two different categories. Two-category sets led to increased memory capacity only when upright faces were included. Capacity for face-only sets never exceeded their nonface counterparts, and the advantage for two-category sets when faces were one of the categories disappeared when inverted faces were used. These results suggest that two-category sets which include faces are advantaged in working memory but that faces alone do not lead to a memory capacity advantage.

Were you paying attention to where you looked? The role of executive working memory in visual search.

Recent evidence has indicated that performing a working memory task that loads executive working memory leads to less efficient visual search (Han & Kim, 2004). We explored the role that executive functioning plays in visual search by examining the pattern of eye movements while participants performed a search task with or without a secondary executive working memory task. Results indicate that executive functioning plays two roles in visual search: the identification of objects and the control of the disengagement of attention.

Are changes in semantic and structural information sufficient for oculomotor capture?

The abrupt onset of objects often involuntarily captures attention (J. Jonides & S. Yantis, 1988) and the eyes (J. Theeuwes, A. F. Kramer, S. Hahn, & D. Irwin, 1998). The new-object hypothesis proposes that the appearance of something new (new semantic and structural information and/or spatiotemporal newness), not the accompanying low-level perceptual transients, causes an involuntary reorienting of attention (S. Yantis & A. P. Hillstrom, 1994). We investigated whether semantic and structural changes alone are sufficient to capture the eyes as strongly as abrupt onsets do. Observers moved their eyes to a target object while another object either onset or smoothly and quickly morphed. If semantic and structural changes are sufficient to capture the eyes, morphs should capture the eyes as strongly as onsets do. Results show that morphs were not fixated first as often as onsets. These findings indicate that new semantic and structural information alone is far less effective at capturing the eyes as onsets.

What we remember affects how we see: spatial working memory steers saccade programming.

Relationships between visual attention, saccade programming, and visual working memory have been hypothesized for over a decade. Awh, Jonides, and Reuter-Lorenz (Journal of Experimental Psychology: Human Perception and Performance 24(3):780-90, 1998) and Awh et al. (Psychological Science 10(5):433-437, 1999) proposed that rehearsing a location in memory also leads to enhanced attentional processing at that location. In regard to eye movements, Belopolsky and Theeuwes (Attention, Perception & Psychophysics 71(3):620-631, 2009) found that holding a location in working memory affects saccade programming, albeit negatively. In three experiments, we attempted to replicate the findings of Belopolsky and Theeuwes (Attention, Perception & Psychophysics 71(3):620-631, 2009) and determine whether the spatial memory effect can occur in other saccade-cuing paradigms, including endogenous central arrow cues and exogenous irrelevant singletons. In the first experiment, our results were the opposite of those in Belopolsky and Theeuwes (Attention, Perception & Psychophysics 71(3):620-631, 2009), in that we found facilitation (shorter saccade latencies) instead of inhibition when the saccade target matched the region in spatial working memory. In Experiment 2, we sought to determine whether the spatial working memory effect would generalize to other endogenous cuing tasks, such as a central arrow that pointed to one of six possible peripheral locations. As in Experiment 1, we found that saccade programming was facilitated when the cued location coincided with the saccade target. In Experiment 3, we explored how spatial memory interacts with other types of cues, such as a peripheral color singleton target or irrelevant onset. In both cases, the eyes were more likely to go to either singleton when it coincided with the location held in spatial working memory. On the basis of these results, we conclude that spatial working memory and saccade programming are likely to share common overlapping circuitry.

The interaction between memorized objects and abrupt onsets in oculomotor capture.

Recent evidence has been found for a source of task-irrelevant oculomotor capture (defined as when a salient event draws the eyes away from a primary task) that originates from working memory. An object memorized for a nonsearch task can capture the eyes during search. Here, an experiment was conducted that generated interactions between the presence of a memorized object (a colored disk) with the abrupt onset of a new object during visual search. The goal was to compare memory-driven oculomotor capture to oculomotor capture caused by an abrupt onset. This has implications for saccade programming theories, which have little to say about saccades that are influenced by object working memory. Results showed that memorized objects capture the eyes at nearly the same rate as abrupt onsets. When the abrupt onset and a memorized color coincide in the same object, this combination leads to even greater oculomotor capture. Finally, latencies support the competitive integration model: Shorter saccade latencies were found when the memorized color combined with the onset captured the eyes, as compared to either color or onset only. Longer latencies were also found when the color and onset occurred in the same display but were spatially separated.