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The metastasis-invasion cascade describes the series of steps required for a cancer cell to successfully spread from its primary tumor and ultimately grow within a secondary organ. Despite metastasis being a dynamic, multistep process, most omics studies to date have focused on comparing primary tumors to the metastatic deposits that define end-stage disease. This static approach means we lack information about the genomic and epigenomic changes that occur during the majority of tumor progression. One particularly understudied phase of tumor progression is metastatic colonization, during which cells must adapt to the new microenvironment of the secondary organ. Through temporal profiling of chromatin accessibility and gene expression in vivo, we identify dynamic changes in the epigenome that occur as osteosarcoma tumors form and grow within the lung microenvironment. Furthermore, we show through paired in vivo and in vitro CRISPR drop-out screens and pharmacological validation that the upstream transcription factors represent a class of metastasis-specific dependency genes. While current models depict lung colonization as a discrete step within the metastatic cascade, our study shows it is a defined trajectory through multiple epigenetic states, revealing new therapeutic opportunities undetectable with standard approaches.

Individual neuronal, signal transduction, and regulatory pathways often control multiple stochastic downstream actuators, which raises the question of how coordinated response to a single input can be achieved when individual actuators fluctuate independently. In Escherichia coli , the bacterial chemotaxis pathway controls the activity of multiple flagellar motors to generate the run-and-tumble motion of the cell. High-resolution microscopy experiments have identified the key conformational changes adopted by individual flagella during this process. By incorporating these observations into a stochastic model of the flagellar bundle, we demonstrate that the presence of multiple motors imposes a trade-off on chemotactic performance. Multiple motors reduce the latency of the response below the time scale of the stochastic switching of a single motor, which improves performance on steep gradients of attractants. However, the uncoordinated switching of multiple motors interrupts and shortens cell runs, which thereby reduces signal detection and performance on shallow gradients. Remarkably, when slow fluctuations generated by the adaptation mechanism of the chemotaxis system are incorporated in the model at levels measured in experiments, the chemotactic sensitivity and performance in shallow gradients is partially restored with marginal effects for steep gradients. The noise is beneficial because it simultaneously generates long events in the statistics of individual motors and coordinates the motors to generate a long tail in the run length distribution of the cell. Occasional long runs are known to enhance exploration of random walkers. Here we show that they have the additional benefit of enhancing the sensitivity of the bacterium to very shallow gradients.

The chemotaxis signalling network in Escherichia coli that controls the locomotion of bacteria is a classic model system for signal transduction. This pathway modulates the behaviour of flagellar motors to propel bacteria towards sources of chemical attractants. Although this system relaxes to a steady state in response to environmental changes, the signalling events within the chemotaxis network are noisy and cause large temporal variations of the motor behaviour even in the absence of stimulus. That the same signalling network governs both behavioural variability and cellular response raises the question of whether these two traits are independent. Here, we experimentally establish a fluctuation-response relationship in the chemotaxis system of living bacteria. Using this relationship, we demonstrate the possibility of inferring the cellular response from the behavioural variability measured before stimulus. In monitoring the pre- and post-stimulus switching behaviour of individual bacterial motors, we found that variability scales linearly with the response time for different functioning states of the cell. This study highlights that the fundamental relationship between fluctuation and response is not constrained to physical systems at thermodynamic equilibrium but is extensible to living cells. Such a relationship not only implies that behavioural variability and cellular response can be coupled traits, but it also provides a general framework within which we can examine how the selection of a network design shapes this interdependence.

The conductance of molecular junctions, formed by breaking gold point contacts dressed with various thiol functionalized organic molecules, is measured at 293 K and at 30 K. In the presence of molecules, individual conductance traces measured as a function of increasing gold electrode displacement show clear steps below the quantum conductance steps of the gold contact. These steps are distributed over a wide range of molecule-dependent conductance values. Histograms constructed from all conductance traces therefore do not show clear peaks either at room or low temperatures. Filtering of the data sets by an objective automated procedure only marginally improves the visibility of such features. We conclude that the geometrical junction to junction variations dominate the conductance measurements.

Bacterial chemotaxis systems are as diverse as the environments that bacteria inhabit, but how much environmental variation can cells tolerate with a single system? Diversification of a single chemotaxis system could serve as an alternative, or even evolutionary stepping-stone, to switching between multiple systems. We hypothesized that mutations in gene regulation could lead to heritable control of chemotactic diversity. By simulating foraging and colonization of E. coli using a single-cell chemotaxis model, we found that different environments selected for different behaviors. The resulting trade-offs show that populations facing diverse environments would ideally diversify behaviors when time for navigation is limited. We show that advantageous diversity can arise from changes in the distribution of protein levels among individuals, which could occur through mutations in gene regulation. We propose experiments to test our prediction that chemotactic diversity in a clonal population could be a selectable trait that enables adaptation to environmental variability. DOI: http://dx.doi.org/10.7554/eLife.03526.001, eLife digest Bacterial colonies are generally made up of genetically identical cells. Despite this, a closer look at the members of a bacterial colony shows that these cells can have very different behaviors. For example, some cells may grow more quickly than others, or be more resistant to antibiotics. The mechanisms driving this diversity are only beginning to be identified and understood. Escherichia coli bacteria can move towards, or away from, certain chemicals in their surrounding environment to help them navigate toward favorable conditions. This behavior is known as chemotaxis. The signals from all of these chemicals are processed in E. coli by just one set of proteins, which control the different behaviors that are needed for the bacteria to follow them. Different numbers of these proteins are found in different—but genetically identical—bacteria, and the number of proteins is linked to how the bacteria perform these behaviors. It has been suggested that diversity can be beneficial to the overall bacterial population, as it helps the population survive environmental changes. This suggests that the level of diversity in the population should adapt to the level of diversity in the environment. However, it remains unknown how this adaptation occurs. Frankel et al. developed and combined several models and simulations to investigate whether differences in chemotaxis protein production help an E. coli colony to survive. The models show that in different environments, it can be beneficial for the population as a whole if different cells have different responses to the chemicals present. For example, if a lot of a useful chemical is present, bacteria are more likely to survive by heading straight to the source. If not much chemical is detected, the bacteria may need to move in a more exploratory manner. Frankel et al. find that different amounts of chemotaxis proteins produce these different behaviors. To survive in a changing environment, it is therefore best for the E. coli colony to contain cells that have different amounts of these proteins. Frankel et al. propose that the variability of chemotaxis protein levels between genetically identical cells can change through mutations in the genes that control how many of the proteins are produced, and predict that such mutations allow populations to adapt to environmental changes. The environments simulated in the model were much simpler than would be found in the real world, and Frankel et al. describe experiments that are now being performed to confirm and expand on their results. The model could be used in the future to shed light on the behavior of other cells that are genetically identical but exhibit diverse behaviors, from other bacterial species to more complex cancer cells. DOI: http://dx.doi.org/10.7554/eLife.03526.002

International audience; The tailbud is the posterior leading edge of the growing vertebrate embryo and consists of motile progenitors of the axial skeleton, musculature and spinal cord. We measure the 3D cell flow field of the zebrafish tailbud and identify changes in tissue fluidity revealed by reductions in the coherence of cell motion without alteration of cell velocities. We find a directed posterior flow wherein the polarization between individual cell motion is high, reflecting ordered collective migration. At the posterior tip of the tailbud, this flow makes sharp bilateral turns facilitated by extensive cell mixing due to increased directional variability of individual cell motions. Inhibition of Wnt or Fgf signaling or cadherin 2 function reduces the coherence of the flow but has different consequences for trunk and tail extension. Modeling and additional data analyses suggest that the balance between the coherence and rate of cell flow determines whether body elongation is linear or whether congestion forms within the flow and the body axis becomes contorted.

In many sensory systems, transmembrane receptors are spatially organized in large clusters. Such arrangement may facilitate signal amplification and the integration of multiple stimuli. However, this organization likely also affects the kinetics of signaling since the cytoplasmic enzymes that modulate the activity of the receptors must localize to the cluster prior to receptor modification. Here we examine how these spatial considerations shape signaling dynamics at rest and in response to stimuli. As a model system, we use the chemotaxis pathway of Escherichia coli, a canonical system for the study of how organisms sense, respond, and adapt to environmental stimuli. In bacterial chemotaxis, adaptation is mediated by two enzymes that localize to the clustered receptors and modulate their activity through methylation-demethylation. Using a novel stochastic simulation, we show that distributive receptor methylation is necessary for successful adaptation to stimulus and also leads to large fluctuations in receptor activity in the steady state. These fluctuations arise from noise in the number of localized enzymes combined with saturated modification kinetics between the localized enzymes and the receptor substrate. An analytical model explains how saturated enzyme kinetics and large fluctuations can coexist with an adapted state robust to variation in the expression levels of the pathway constituents, a key requirement to ensure the functionality of individual cells within a population. This contrasts with the well-mixed covalent modification system studied by Goldbeter and Koshland in which mean activity becomes ultrasensitive to protein abundances when the enzymes operate at saturation. Large fluctuations in receptor activity have been quantified experimentally and may benefit the cell by enhancing its ability to explore empty environments and track shallow nutrient gradients. Here we clarify the mechanistic relationship of these large fluctuations to well-studied aspects of the chemotaxis system, precise adaptation and functional robustness., Author Summary To navigate their environments, organisms must remain sensitive to small changes in their surroundings while adapting to persistent conditions. Bacteria travel by performing a random walk biased toward nutrients and away from toxins. The decision of a bacterium to continue in a given direction or to reorient is controlled by the state of its chemoreceptors. Chemoreceptors assemble into large polar clusters, an arrangement required for the amplification of small stimuli. We investigate how this organization affects the kinetics of the enzymatic reactions through which the receptors adapt to persistent stimuli. We show that clustering can lead to large fluctuations in the state of the receptors, which have been observed in Escherichia coli and may aid in the navigation of weak stimulus gradients and the exploration of sparse environments. Additionally, we show that these fluctuations can occur around a mean receptor state robust to changes in the numbers of the adaptation enzymes. Since enzyme expression levels vary across a population, this feature ensures a high proportion of functional cells. Our study clarifies the relation between fluctuations, adaptation, and robustness in bacterial chemotaxis and may inform the study of other biological systems with clustered receptors or similar enzyme-substrate interactions.

Using in vitro and in vivo assays, we define a network of Her/Hes dimers underlying transcriptional negative feedback within the zebrafish segmentation clock. Some of the dimers do not appear to be DNA-binding, whereas those dimers that do interact with DNA have distinct preferences for cis regulatory sequences. Dimerization is specific, with Hes6 serving as the hub of the network. Her1 binds DNA only as a homodimer but will also dimerize with Hes6. Her12 and Her15 bind DNA both as homodimers and as heterodimers with Hes6. Her7 dimerizes strongly with Hes6 and weakly with Her15. This network structure engenders specific network dynamics and imparts greater influence to the Her7 node. Computational analysis supports the hypothesis that Her7 disproportionately influences the availability of Hes6 to heterodimerize with other Her proteins. Genetic experiments suggest that this regulation is important for operation of the network. Her7 therefore has two functions within the zebrafish segmentation clock. Her7 acts directly within the delayed negative feedback as a DNA-binding heterodimer with Hes6. Her7 also has an emergent function, independent of DNA binding, in which it modulates network topology via sequestration of the network hub.

We have developed a unique micromechanical method to extract extremely thin graphite samples. Graphite crystallites with thicknesses ranging from 10 - 100 nm and lateral size $\sim$ 2 $\mu$m are extracted from bulk. Mesoscopic graphite devices are fabricated from these samples for electric field dependent conductance measurements. Strong conductance modulation as a function of gate voltage is observed in the thinner crystallite devices. The temperature dependent resistivity measurements show more boundary scattering contribution in the thinner graphite samples., Comment: 3 pages, 3 figures included, submitted to Appl. Phys. Lett

Experimental psychotic children, 9 boys and 3 girls (mean age=66.9 mos.) were compared with 5 non-psychotic subnormal boys (mean age=72 mos.) in body imitation, motor-object imitation and spontaneous object use and also for influence of task class and difficulty using each subject's adaptive age as referant. Psychotic children did poorly in body, better in motor-object imitation and best in object use. Controls achieved higher body and motor-object imitation scores. Task difficulty equally affected the performances of subjects and controls. Body imitation failure, traced to neurophysiological deficiencies, combined with severe central language disorders in the psychotic group, precluded non-verbal and verbal interpersonal communication. It is suggested that during early life, psychotic children, unlike the subnormal who have less severe visual-motor or language processing handicaps, cannot use parents' help to interpret environment.

Five diagnostic systems designed to differentiate infantile autism and early childhood schizophrenia were compared by deriving scores on 44 children referred consecutively to the same clinical center. While the autistic scales devised by Rimland, Polan and Spencer, Lotter, and the British Working Party correlated significantly, the degree of correspondence (35%) indicated that several children obtained high autistic scores in one system but low scores in another. The BWP's term “schizophrenia” has more correspondence with the term “autism” used by others than with Rimland's “schizophrenia.” In the DeMyer-Churchill categorical system (early schizophrenia, primary autism, secondary autism, and non-psychotic subnormal), “primary autism” most resembles Rimland's concept of infantile autism as measured by his E-1 version. All other systems differentiate psychotic from non-psychotic children but do not distinguish any of the psychotic subgroups.

Infant care practices and innate activity are compared in a study of 96 families representing 33 autistic and early schizophrenic, 33 matched normal, and 30 subnormal children. Data from objectively rated interviews indicated that parents of psychotic and normal children were alike in infant care practices, total stimulation scores, and factor analytically derived scores. Parents of subnormals were the least stimulating and coldest, providing fewer contacts and less physical freedom to the infants than the other parents. Normal infants were more active but autistic and subnormal did not differ on any infant rating item. Infant factor scores showed that autistic and subnormal infants were most alike in the alertness factor, both below normal infants. Social factors did not differ. Brain damage indices were highest in subnormal and lowest in normal, all differing significantly. Results are discussed and interpreted to substantiate criticism of theories that attribute autism to parental or organic-environmental causation during infancy.

Due to etiological implications, parental intellectuality defined by 29 descriptive rating scales was compared in 96 families. Parents represented 33 autistic and schizophrenic children, 33 matched normals, and 30 subnormals. Data was obtained from objectively rated interviews, WAIS and other scales. All groups were alike in characteristics they sought in spouses and children, premarital interests, reading preferences, and life style. With the child's age, sex, ordinal position, race, religion and SES held constant, only one significant difference was found between parents of normal and autistic children. The latter emphasized academic success less in autistic than matched normals in their children. All parents of deviant children desired improvement in speech and relatedness, realistically deemphasizing intellectual achievement. Fathers' verbal IQs were significantly higher for autistic than subnormal groups. Parents of normals were significantly younger at child's birth, an unexpected finding implying a neurological link between autism and subnormality.