Michael Miles - US grants

clearance rate; anticonvulsants; pharmacokinetics; human puberty; adolescence (12-20); age difference; carbamazepine; valproate; phenytoin; gender difference; clinical research; human subject;

DESCRIPTION (provided by applicant): Acute ethanol exposure stimulates brain reward pathways. Upon chronic exposure, neural adaptations occur which generate new and long lasting behaviors such as tolerance, sensitization, and craving. We, and others, have hypothesized that stepwise changes in gene expression are an important part of the molecular mechanisms underlying experience-dependent neural adaptations leading to ethanol addiction. Studies in humans and animals have shown that acute behavioral responses to ethanol have predictive value in terms of the risk for excessive ethanol consumption and alcoholism. Here we hypothesize that changes in brain gene expression occurring with acute ethanol are an important part of the interplay between acute behavioral responses to ethanol and the risks for abusive drinking behavior. We propose to use high-density oligonucleotide arrays to provide a non-biased and a near genomic-level analysis of gene expression in brain areas related to the mesolimbocortical dopamine reward pathway. Our laboratory has developed extensive experience in the use of high-density oligonucleotide arrays for the study of changes in brain gene expression resulting from exposure to drugs of abuse such as ethanol or cocaine. We will characterize the time course and dose response of expression changes occurring in four different brain regions following acute ethanol exposure. Results will be contrasted in two different inbred lines of mice, C57BL/6J (B6) and DBA/2J (D2), which have markedly divergent responses to acute ethanol and tendencies for ethanol consumption. A comprehensive data analysis approach will be utilized including multivariant studies such as hierarchical clustering and self-organizing maps, together with a high throughput mining of biological databases. The goal will be to identify clusters of genes with expression patterns correlating with the differing behavioral responses of B6 and D2 mice. Such expression patterns may identify cellular functions critical to the neurobiology of acute ethanol action. The second specific aim will study the pharmacology of expression changes occurring with acute ethanol. In particular, we will characterize whether agents known to alter acute ethanol behavioral responses and ethanol drinking behavior also modulate specific gene expression profiles correlated with acute ethanol actions. Finally, we will determine whether prior experience with ethanol, as seen with locomotor sensitization, alters expression profiles elicited by acute ethanol exposure. This will identify acute ethanol-induced expression patterns that are potentially important for long lasting neuroadaptive events occurring with repeated ethanol exposure. All of the data produced in these studies will be rapidly made available to other investigators through a web site we have already established for posting array results. Such information dissemination is crucial for the "hypothesis generating" capacity of the massively parallel and non-biased observations generated by DNA arrays. In summary, this work will greatly increase our understanding of brain molecular responses to ethanol that are important for either acute behavioral responses or long-term maladaptive behaviors as seen in alcoholism. These findings might generate novel insights for the treatment of alcoholism.

DESCRIPTION (provided by applicant): Although much has been learned about molecular sites of action for ethanol, there remains few effective treatments for alcoholism. Naltrexone (NTX) reduces recidivism and ethanol consumption in alcoholics. NTX, an un-selective opioid antagonist, has also been shown to decrease ethanol drinking behavior in animal models, including blocking increased ethanol drinking in a model of relapse drinking, the ethanol deprivation effect (EDE). The molecular mechanism(s) for these responses are not entirely understood. We hypothesize that by studying genome-wide gene expression patterns associated with naltrexone action, EDE and naltrexone effects on EDE, we might gain novel insight into mechanisms relevant to relapse drinking behavior. In this project high-density oligonucleotide arrays will first be used to characterize gene expression patterns evoked by NTX in naive C57BL/6 mice. Ventral tegmental area, nucleus accumbens and medial prefrontal cortex brain regions in C57BL/6 mice will be studied. Expression profiles of NTX will also be compared to those from two other agents that decrease ethanol drinking or the EDE, acamproste and MPEP, an inhibitor of the mGluR5 glutamate receptor. Aim two will then use arrays to study action of NTX on gene expression patterns evoked by ethanol-deprivation in a 2-bottle choice model of ethanol self administration. Through data mining the combined expression patterns related to NTX action in aims 1-2, we will then characterize particular candidate genes in regard to cellular patterns of their expression changes. In Aim 3, candidate genes will be evaluated for their role in ethanol drinking or the EDE in a 2-bottle choice model. Pharmacological or genetic (viral vectors, antisense oligonucleotides) means will be used to alter the expression of candidate genes prior to behavioral testing. These studies should provide novel insight into the mechanisms of the EDE and mechanisms of NTX action in altering ethanol drinking behavior. Together, these findings may identify novel targets for therapeutic intervention in alcoholism.

[unreadable] DESCRIPTION (provided by applicant): he Informatics and Analysis Core (BINFO) will provide several distinct functions to serve INIA-Stress investigators and projects. Because of the scope and diversity of INIA-Stress research projects, the general role of the BINFO will be to provide standardized research support and key integrative functions among the research projects. The BINFO will provide support for experimental design, advanced statistical analysis, data warehousing or database storage, and advanced bioinformatics analysis, including development or adaptation of novel algorithms for analysis of gene networks. The BINFO encompasses investigators at four different institutions (Virginia Commonwealth University, Oak Ridge National Laboratory, University of Tennessee at Knoxville and Vanderbilt University) and will be divided into subcores based upon different disciplines and involve expertise from multiple sites. The subcores and related aims of the BINFOC are: 1) Provide a web-based information system (www.iniastress.org) that will have both public and INIA-Stress restricted functions; 2) Provide advanced experimental design and statistical support for all INIA-Stress projects; 3) Provide algorithms and assistance for gene-network annotation (INIAGestalt) analysis and gene network integration across INIA-Stress datasets; and 4) Provide expertise and tools for experimental design, analysis and database management of microarray experiments for INIA-Stress. Altogether the BINFO will provide a forum for both data integration and advanced data analysis across all the primary projects of this proposal. In addition, the tools and resources provided by the BINFO will provide service to the alcohol research community and other biomedical researchers. [unreadable] [unreadable] [unreadable]

[unreadable] DESCRIPTION (provided by applicant): Recent studies in our laboratory on ethanol 2-bottle choice drinking with C57BL/6 inbred mice have documented exceedingly consistent, substantial individual variation in daily drinking behavior. This individual variation in ethanol drinking is modified by environmental factors, including formation of social hierarchy through group housing with non-siblings. This variation in drinking is not seen for saccharin or total fluid intake. Initial microarray studies done on individual animals have identified expression patterns highly correlated with this drinking behavior. It is our hypothesis that stress (isolation, social hierarchy) has created long-lasting influences altering drinking behavior in these genetically identical animals. This offers a unique opportunity to finely dissect molecular mechanisms underlying difference in drinking behavior generated by environmental influences, particularly in regard to social interactions. We predict our results will partially overlap with drinking behavior-related gene networks identified by "traditional" genetic approaches, as well as identify novel modulators of drinking behavior not discernable in studies on populations. We will conduct five specific aims. Aim 1 will further characterize our behavioral model to determine the stability of the variation in individual drinking behavior induced by group housing. Aim 2 will characterize whether individual variation in drinking behavior correlates with behavioral or neurochemical measures of anxiety and stress. These studies will include assessment of plasma corticosterone levels and brain regional levels of corticotropin releasing factor (CRF) and the neurosteroid, allopregnanaolone. Aim 3 will use whole genome microarray studies to identify gene expression patterns correlating with drinking behavior or social stressors such as group housing or isolation housing. Aim 4 will refine microarray data using bioinformatic resources through the INIA-Stress bioinformatics core and collaborations with Drs. Chester, Langston and Williams. Candidate genes will be selected through a multivariate ranking scheme and expression verified by Q-rtPCR, Western blotting and immunocytochemistry. Candidates selected from our microarray studies will also be used to determine if the same genes show correlations with stress or drinking behavior in mouse samples from Drs. Biggio (Project 11) and primates (Dr. Grant, Project 3). In Aim 5, these genes will be used to predict drinking behavior or drinking responses to social stress in mouse lines derived from the BXD recombinant inbred panel or othe mouse resources from this INIA. Finally, these candidate genes will be behaviorally verified by generating null mouse lines from genetrap ES cells (via the Knockout Core, Dr. Delpire) or using viral vectors to deliver gene constructs to target brain resions followed by behavioral studies. These studies might thus identify genes or networks of genes that cause human alcohol consumption to vary across individuals, leading some to excessive alcohol intake and risk for alcoholism. [unreadable] [unreadable] [unreadable]

The objective of this Small Grant for Exploratory Research (SGER) project is to study the capability of a new solid state joining process, termed friction bit joining, to weld dissimilar combinations of aluminum, magnesium and steel automotive sheets. The automotive industry and the US Government are aggressively pursuing use of lightweight materials such as Mg alloys and Al alloys in auto body structures for improved vehicle fuel efficiency. High-volume production vehicles would require use of a variety of engineering materials, ranging from ultra high-strength steels (UHSS), Al alloys, Mg alloys and others to meet cost and performance objectives. The combination of lightweight materials with widely acceptable structural materials such as UHSS presents a number of technical challenges in automotive body-in-white assembly. One of them is joining different materials to form an integrated structure component which meets design and performance requirements. The current work will study the welding process from a fundamental viewpoint, including the effect of process conditions on metallurgical bond properties of dissimilar alloy joints. The friction bit joining process has been shown to be capable of achieving high levels of strength in dissimilar alloy joints of DP 980 steel and 5754 aluminum alloys during a preliminary study. This combination of ultra high strength steel and a soft aluminum alloy would be difficult or impossible to join using conventional spot welding or even self piercing riveting.

If this new process is shown to be successful in bonding dissimilar metal alloys where strength levels are very different, it will promote greater use of light metals in automotive and other transportation structures, allowing for reduced weight and improved fuel efficiency.

Studies in both humans and animal models show that acute behavioral responses to ethanol have predictive value regarding risk for long term ethanol drinking behavior. In rodent models, locomotor activation and anxiolytic-like activity are two widely documented acute behavioral responses to ethanol. Stress, anxiety and effects of ethanol or ethanol-withdrawal on anxiety have been proposed as important factors in the genesis of alcoholism and recidivism. Recent results in our laboratory have identified significant quantitative trait loci (QTLs) for acute ethanol anxiolytic-like activity using the light-dark box transition model of anxiety across 33 BXD recombinant inbred mouse strains. Simultaneously, we have generated microarray datasets from the same saline or acute ethanol-treated BXD lines, for prefrontal cortex and nucleus accumbens. Recent studies suggest that combining QTL analysis of gene expression and behavioral can improve identification of candidate genes for behavioral QTLs. Such "expression genetics" has the power to identify networks of highly correlated gene expression patterns. Correlating genetic expression networks with behavioral QTLs could provide evidence for the functional relevance of such expression networks and candidate mechanisms underlying complex traits. This developmental project will furthermore use the power of the overall experimental design of the VCU-ARC to prioritize gene expression networks linked to acute ethanol behavioral responses in mice. We will: 1) Complete microarray studies for ventral tegmental area across 33 BXD Rl lines +/- ethanol (1.8 g/kg x 4 hours);2) Identify gene expression networks correlated with behavioral measures of acute ethanol effects on locomotor activity and anxiety-like behavior;and 3) Through potential cross-species comparison (Projects 2, 3 and Pilot 1) and bioinformatics prioritization (Core 2) of PCR-verified expression changes, we will select a limited number of candidates for direct verification studies in mice. The latter will use behavioral testing of existing null mutation mouse lines or animals treated with viral-vector gene delivery. We expect this to provide a novel validation of gene networks correlated with the acute behavioral responses to ethanol. Furthermore, our mouse gene targeting and behavioral assays will provide resources for studies in Projects 2,3 and Pilot 1 of this Center. Together we expect a novel synergy of these cross-species studies on gene networks responding to acute ethanol.

Studies in both humans and animal models show that acute behavioral responses to ethanol have predictive value regarding risk for long term ethanol drinking behavior. In rodent models, locomotor activation and anxiolytic-like activity are two widely documented acute behavioral responses to ethanol. Stress, anxiety and effects of ethanol or ethanol-withdrawal on anxiety have been proposed as important factors in the genesis of alcoholism and recidivism. Recent results in our laboratory have identified significant quantitative trait loci (QTLs) for acute ethanol anxiolytic-like activity using the light-dark box transition model of anxiety across 33 BXD recombinant inbred mouse strains. Simultaneously, we have generated microarray datasets from the same saline or acute ethanol-treated BXD lines, for prefrontal cortex and nucleus accumbens. Recent studies suggest that combining QTL analysis of gene expression and behavioral can improve identification of candidate genes for behavioral QTLs. Such expression genetics has the power to identify networks of highly correlated gene expression patterns. Correlating genetic expression networks with behavioral QTLs could provide evidence for the functional relevance of such expression networks and candidate mechanisms underlying complex traits. This developmental project will furthermore use the power of the overall experimental design of the VCU-ARC to prioritize gene expression networks linked to acute ethanol behavioral responses in mice. We will: 1) Complete microarray studies for ventral tegmental area across 33 BXD Rl lines +/- ethanol (1.8 g/kg x 4 hours); 2) Identify gene expression networks correlated with behavioral measures of acute ethanol effects on locomotor activity and anxiety-like behavior; and 3) Through potential cross-species comparison (Projects 2, 3 and Pilot 1) and bioinformatics prioritization (Core 2) of PCR-verified expression changes, we will select a limited number of candidates for direct verification studies in mice. The latter will use behavioral testing of existing null mutation mouse lines or animals treated with viral-vector gene delivery. We expect this to provide a novel validation of gene networks correlated with the acute behavioral responses to ethanol. Furthermore, our mouse gene targeting and behavioral assays will provide resources for studies in Projects 2,3 and Pilot 1 of this Center. Together we expect a novel synergy of these cross-species studies on gene networks responding to acute ethanol.

DESCRIPTION (provided by applicant): The challenges and goals for the next phase of genetic research on alcohol dependence (AD) will be to i) confirm candidate genes, ii) to understand the mechanism(s) of action for individual genes in AD and iii) to use the molecular and genetic information to identify potential targets for novel interventions in AD. As a strategy to approach these complex goals, we have formulated three major organizing hypotheses that integrate and direct the individual components of this Developmental (P20) application. These include: 1. A focus on gene networks rather than individual genes which will provide mechanistic information, target identification and cross-species validation of molecular events affecting alcohol-related behaviors and risk for AD. 2. A bi-directional cross-species gene discovery and validation scheme that can provide both powerful confirmation of candidate genes and mechanistic information. Our planned studies in mice (Project 1), humans (Project 2, Pilot 2), C. elegans (Project 3), and D. melanogaster (Pilot 1) will be mutually reinforcing. 3. Initial sensitivity and acute tolerance to ethanol are phenotypes with broad cross-species experimental applicability and validated relevance to AD. For this project, for which we request four years of support, we outline a series of three developmental and two pilot projects which form a highly integrated and novel approach that implement the hypotheses outlined above. An Administrative Core and Bioinformatics Core will provide needed support across projects. The Specific Aims of individual projects will include developmental aspects that seek to broaden our scientific base, increase integration between Center components and extend our experimental models to eventually include additional behavioral phenotypes such as acute and chronic tolerance and dependence. We expect novel contributions to the field of alcohol research from our cross-species analysis of acute ethanol effects.

This research project will build a framework for the design and manufacture of metal microstructures containing fewer damage-sensitive features, with subsequent improved ductility and formability. An enabling technology, Defect Detection Microscopy (DDM), will be developed in order to detect large numbers of defect sites in deformed polycrystalline materials in reasonable experimental times. Structure parameters (recovered via DDM) in the regions of critical defects will be correlated with defect type to determine robust Failure Initiation Parameters (FIPs). Once these correlations are known, an inverse approach, already developed for microstructure sensitive design of defect insensitive elastic and plastic properties, will be applied to the framework to improve defect-sensitive properties. This work will be undertaken in collaboration with General Motors Research Laboratory, and will focus on improved ductility and formability of magnesium for the manufacture of lightweight automotive structures.
The introduction of lightweight materials into the US auto industry is a key national objective that will benefit significantly from this work. Current production methods for fabricating magnesium autobody panels require high temperature processing, preventing the use of magnesium as a viable lightweight alloy for high-volume automobile production. If the aims of this project are met, DDM will not only open the way for cost effective, low temperature, application of magnesium to improve vehicle fuel efficiency in the auto industry, but will also serve as an enabling technology in the study and development of a much broader range of damage-sensitive material applications (such as toughness and fatigue) critical to US industry in general. The proposed interdisciplinary activity also brings together expertise from several traditional fields including mechanical engineering, manufacturing science, engineering design, materials science, and applied mathematics. This will have a significant impact on the development of skilled human resources in emerging science and advanced technology fields.

The research objective of this award is to test the hypothesis that joint bond area in friction stir spot welding of ultra-high strength steel is determined largely by interface pressure, interface temperature, and weld time. Welding speeds in the range of 5,000 to 10,000 revolutions per minute will be considered, and the welding process will be constrained to a maximum vertical welding load of eight kilonewtons, with a goal of achieving a desired level of joint strength. A novel approach to modeling friction stir spot welding will be used within a Lagrangian finite element framework, where the joint interface will be represented virtually and where a proposed relationship between joint contact pressures, temperatures, and weld time will be used to estimate bond area. Archard's law will be used within the model to estimate tool wear. An experimental program will measure joint strength and tool wear over time, while microstructural evaluation of the joints will provide insight into the effect of tool wear on bonding mechanisms.

If successful, this project will enable the use of friction stir spot welding for joining ultra-high strength steel sheet in many industrial settings. The modeling approach will provide increased understanding of the role of tool design and process conditions on joint strength and tool life. Finding a solution to spot joining of ultra-high strength steels is important because these alloys have the potential to make car frames lighter and stiffer, improving vehicle fuel economy and reducing carbon emissions. The widespread use of ultra-high strength steel is currently limited by brittle weld microstructures produced by traditional resistance spot welding.

DESCRIPTION (provided by applicant): An underlying premise of the INIA-Stress consortium is that progression to abusive ethanol consumption is, at least in part, accompanied and perhaps caused by alterations in an organism's response to stress, including the stress of excessive ethanol intake/withdrawal. We propose that changes in brain gene expression networks are an important part of allostatic mechanisms leading to progressive ethanol consumption and aberrant responses to stress. We have previously used genetic and genomic approaches across brain regions of BXD recombinant inbred panel to define robust gene networks regulated by acute ethanol and relate these to ethanol behaviors, particularly regarding responses to stress. We have also identified significant overlap in expression responses to acute ethanol in mice and altered gene expression patterns seen in a primate model of excessive ethanol intake (SIP) developed by Dr. Grant, the PI of the INIA-Stress consortium. Furthermore, very recent pilot array studies in BXD mice exposed to multiple cycles of the chronic intermittent ethanol vapor (CIE) model of excessive ethanol consumption have identified remarkable homology with results from acute ethanol exposure and our data from cynomolgus macaque. Those studies have generated gene networks that allow testing initial major hub genes for their possible role in modifying ethanol consumption and response to stress in the CIE model. For example, we recently identified Gsk3p as an important regulator of ethanol consumption and withdrawal-induced anxiety, using AAV viral vector gene delivery studies. Based on these findings, we propose the following Organizing Hypothesis: Altered ethanol drinking and stress/endocrine phenotypes in the mouse CIE and monkey SIP models result from (and cause) adaptive responses in brain gene expression networks, resulting in a new allostatic set point. The aims of this project will define new gene networks underlying allostatic changes in the CIE and monkey SIP models by expression profiling of CIE treatment across the BXD mouse panel and Rhesus Macaque samples of Dr. Grant (Project 1), co-analysis of results with RNA-Seq data of Dr. Williams' Project 10, and testing of candidates, including Gsk3P, using viral vector gene delivery. PUBLIC HEALTH RELEVANCE: The studies proposed here can identify novel targets for future therapeutic development and contribute to our overall understanding of the neurobiology underlying the interaction between stress and ethanol during the transition to excessive ethanol consumption. Studies on one candidate gene already identified, Gsk3?, hold direct promise for future medication development.

DESCRIPTION (provided by applicant): Alcohol-related disorders impose a substantial burden on society with far-reaching health consequences. The identification of novel genes and genetic pathways that influence alcohol-related behaviors will facilitate the development of new therapeutic interventions for alcoholism and other forms of alcohol abuse. In this project we will investigate genes and genetic pathways that have novel influences on ethanol behavior. Molecular-genetic information from this project should ultimately lead to better diagnosis, risk determination and treatment of alcohol-related disorders in humans. This project focuses on Clic4/Clic as a novel mouse/Drosophila gene that affects behavioral responses to ethanol. Preliminary studies indicate that this gene influences ethanol-related behavior in both fruit flies (Drosophila) and mice, suggesting that it has a conserved role in ethanol action. To further characterize Clic4/Clic and its associated molecular mechanisms in ethanol behavior, we have developed a coordinated study in Drosophila and mice. Using the Drosophila model, this project will identify the tissue site of Clic action (Aim 1), define the temporal requirements for Clic (Ai 2) and delineate molecular-genetic mechanisms of Clic function (Aim 3). Using the mouse model, this project will further characterize ethanol regulation of Clic4 in the brain (Aim 4A), characterize the role of mammalian Clic4 in drinking and other ethanol behaviors (Aim 4B), characterize downstream molecular responses to altered Clic4 expression (Aim 4C), and investigate the role of a focused set of molecular partners implicated in Clic4 action (Aim 4D). This project draws on the complementary expertise of two independent laboratories directed by PIs Grotewiel (fly) and Miles (mouse) and is designed to have several major points of integration. Clic4/Clic was originally implicated as a candidate gene for ethanol behavior by a series of analyses by the Miles laboratory on gene expression, linkage and association data. Subsequent genetic analysis of Clic in the fly by the Grotewiel laboratory made Clic4 a high priority locus for ethanol behavioral studies in the mouse (described as preliminary data). These studies come together to rationally support a more extensive investigation of how Clic/Clic4 influences ethanol responses in flies (Aims 1 and 2) and mice (Aims 4A and 4B). Furthermore, additional studies on ethanol-responsive genes in the mouse (Aims 4A and 4C) are now informing the design of experiments in flies that will investigate mechanisms of Clic action (Aim 3). The results of the Drosophila studies in Aim 3 will in turn guide the design of experiments in mice on mechanisms for Clic4 in mammalian ethanol behavior (Aim 4D). The deliberate cross-species integration in this project, implemented within a collaborative framework between the Miles and Grotewiel laboratories, will drive a vigorous genetic investigation of conserved mechanisms underlying ethanol behavior. PUBLIC HEALTH RELEVANCE: Alcoholism and other forms of alcohol abuse lead to major health problems. This project will use the fruit fly and the mouse in experiments that will investigate how genes contribute to the effects of ethanol in the brain. The long-term goal of this project is to generate information about genes that will lead to new treatments for alcoholism and alcohol abuse.

DESCRIPTION (provided by applicant): Despite evidence of strong genetic contributions to the etiology of smoking initiation (SI) and nicotine dependence (ND), we are far from identifying the specific genetic basis of individual susceptibility to ND. This project - to deepen our understanding of how genes contribute to risk for nicotine dependence (ND) - has arisen as an effort to utilize maximally the relatively unique and complementary set of skill of this group of investigators in complex human genetics, animal genetics and nicotine pharmacology. We will validate these putative risk genes using a two-step approach: replication in other human samples and the demonstration in animal models of ND that variants in these genes contribute to neurobiological pathways likely involved in ND. We will first identify promising candidate genes for smoking initiation (SI) and ND by data-mining GWA datasets and the selected candidates will be replicated. Using a mouse model of nicotine withdrawal, we will characterize behavioral QTLs relevant for ND using a recently developed expanded BXD RI mouse strain panel, focusing on strains informative for already identified areas of provisional QTLs. This approach allows us to both validate and refine our mapping of the nicotine behavioral QTL. Furthermore, we will identify candidate genes for ND by combining expression and behavioral genetics analyses in these BXD RI mouse strains. Candidate genes/pathways identified and prioritized from human and mouse studies will be validated by pharmacological or genetic manipulations to alter expression or function of candidate genes in mouse brain and determine effects on behavioral responses to in models of nicotine reward and withdrawal.

This Grant Opportunity for Academic Liaison with Industry (GOALI) award supports fundamental research to uncover the mechanisms of sigma phase and chromium carbide formation in friction stir processed austenitic stainless steel. An Eulerian finite element approach will be used to predict temperatures, strains, and strains rates in the stir zone so that fraction recrystallization can be predicted for different tool speeds and feed rates. The modeling work will be combined with experiments to test the hypothesis that sigma phase and chromium carbide formation can be minimized or eliminated when friction stir processing is done under conditions that avoid dynamic recrystallization. Phase identification will be performed on processed specimens by electron backscatter diffraction and by transmission electron microscopy for model validation. Hot cell experiments on irradiated stainless steel will test a second hypothesis that friction stir processing can be done without causing helium cracking in the heat affected zone of the stirred region.

This research will provide fundamental understanding of the role of recrystallization on sigma phase and chromium carbide formation in friction stir processed austenitic stainless steel. A new approach for repairing stress corrosion cracks in nuclear components will emerge from the fundamental research results. As current fusion welding processes cause helium cracking during repair of components that have been irradiated for twenty or more years, this new approach has the potential of overcoming a major technical challenge in maintaining our fleet of aging reactors, which is critical to the national energy infrastructure and the economy. Additional societal benefits include the education of students who will be prepared to contribute to the nuclear industry, and an outreach program working with high school teachers on new curriculum development, aimed at encouraging female students to pursue careers in science and engineering.

In order to meet ambitious targets for reduced energy consumption and emissions in modern vehicles, one of the most widely adopted strategies involves the deployment of lightweight structural materials. At less than a quarter of the density of steel, magnesium is a natural contender to replace legacy materials in automotive and other vehicle components. However, despite its desirable physical properties, magnesium accounts for only a small proportion of the make-up of a typical automobile. The reason, in large part, stems from the difficulty with forming the relatively brittle magnesium alloys into complex shapes required for vehicle components. This award supports research into the science underlying micro-scale behavior of magnesium alloys during forming and other deformation operations. By uncovering the relationships between the metal's microstructure and its response to deformation, modified manufacturing processes and improved alloys can be designed to enable widespread use of lightweight magnesium components in vehicular and other weight-sensitive applications. This will be complemented by STEM-oriented outreach activities and external collaborations.

The response of a magnesium component to applied deformation is governed by two atomic-level phenomena: slip and twin activity. Twinning is especially vital to deformation at and near room temperature, due to the difficulty of slip. This project will build upon newly developed microscopy techniques (high-resolution electron backscatter diffraction) to generate snapshots of nano and micro-level structural activity during forming activities. Data mining knowledge extraction techniques will be applied to the resultant huge data sets of twin and slip activity in order to accelerate the discovery of interrelations between microstructure and magnesium deformation mechanics. The data mining will employ a decision-tree type analysis and a neural net approach. The resultant knowledge will be embedded in a meso-scale model of twin / deformation activity as the basis for assessment and design of improved alloys for light-weight structural applications. New insights will emerge into twin development that span various grain-size ranges and critical temperature levels. Key structure parameters will be modeled, including detailed grain-boundary character, accurate local (relative) strain levels, dislocation activity / slip transition temperatures, measures of crystal lattice entropy and other metrics of local heterogeneity. Furthermore, the high-throughput microscopy and data mining advances developed as part of the study will serve as a new framework for accelerated knowledge discovery for constitutive models of deformation in crystalline materials.

NON-TECHNICAL SUMMARY:
The efficiency of modern vehicles critically affects individual and national prosperity, national energy independence, city pollution levels (and related health) and climate change. A key element in deciding this efficiency is vehicle weight. While many ongoing efforts aim at introducing lightweight materials (such as aluminum, magnesium and carbon composites) into automotive structures, the dominant constituent, by far, is still relatively heavy steel. Hence this project focuses on the alternative light-weighting strategy of developing advanced high strength steels with improved formability; this will allow less metal to provide the same structural and safety performance. New and emerging microscopy techniques are combined with simulation to reveal the relationships between microstructure and properties of modern high-strength steels. The resulting insights lead to the deeper levels of understanding and improved models that are required for the nascence of so-called third-generation steels and a step-change in vehicle weight and efficiency. Not only does this research project provide an environment for mentoring graduate and undergraduate students in a mixed academic / industrial setting (via a strong collaboration with General Motors), but high school education is being improved via an outreach program. Teachers from local high schools spend time on the project during the summer months while receiving help in developing new and relevant curriculum components for their classes.

TECHNICAL SUMMARY:
Many studies have been carried out on TRIP (Transformation Induced Plasticity) steels. However, there is still no systematic information concerning the influence of austenite chemistry and microstructure on the resulting transformation to martensite. Such knowledge is critical to the design of texture, morphology and volume fraction of retained austenite in order to enable low-alloy TRIP steels to achieve enhanced strength, formability, and fracture resistance necessary to achieve the desired automotive structure performance. Proposed retained-austenite and martensite identification technologies (based upon emerging electron-backscatter diffraction techniques), along with meso-scale dislocation characterization, are combined with micro-digital image correlation (DIC) to provide a new 'Deformation Microscopy' capability. The resultant framework allows for the assessment of interactions between dislocations and hard phase boundaries, while also precisely capturing the strains at which the various retained austenite islands transform. Knowledge of the fundamental links between microstructure and formability enables the formulation of a meso-scale model for transformation of austenite to martensite, and related deformation phenomena.

A key component in the strategy to lightweight vehicles for reducing harmful emissions involves the introduction of advanced light alloys across a wide spectrum of vehicle components. However, advanced alloys are typically less ductile than their heavier predecessors and are liable to fracture during the shaping and forming operations. On the other hand, various empirical observations have demonstrated that careful selection of strain (deformation) path during the forming process can significantly delay component failure. Current simulation frameworks do not account for key phenomena at the microstructural level needed to analyze and design better forming processes and to guide alloy selection and development for optimal exploitation of current and forthcoming lightweight materials. By combining novel developments in microscopy and modeling, the critical issue to be explored in this Grant Opportunities for Academic Liaison with Industry (GOALI) research project involves interactions between mobile planes of atoms (dislocations) that facilitate shape change of the component, and microstructural interfaces, such as precipitates and grain boundaries. Barriers to dislocation glide cause atomic pileups, and related backstress effects, that are not considered in traditional models, but can potentially be manipulated to improve overall ductility via careful design of strain paths that occur during forming. The research will be integrated into industrial practice by the industrial partner, Aleris, to deliver potentially transformational capabilities in vehicle lightweighting efforts. As a result of this collaboration, the students involved will also gain an understanding of industrial challenges and drivers. Knowledge derived from the research will be integrated into course curricula for graduate and undergraduate students, while a cloud-based App hosting the developed model will be made available to the broader research community via Materials Resources, LLC.

This interdisciplinary project, involving the complementary expertise of two universities and an industrial partner, is driven by the hypothesis that accurate calculation of strain gradients, and related backstress and localization fields, during forming can be used to design strain paths that optimize material ductility, effectively delaying localization/failure in high-strength aluminum (Al) alloy sheets. The team will conceive and implement a novel strain-gradient crystal plasticity finite element model to encapsulate the scientific insights. The model will be guided by a combination of two cutting-edge microstructural techniques that will provide unprecedented detail of the deformation behavior at the relevant length-scale. High-resolution electron backscatter diffraction (HREBSD) will be employed for mapping both geometrically necessary dislocations, accompanying strain gradients, and related backstress for each strain path, while high-resolution digital image correlation (HRDIC) will extract the plastic strain tensor for a complete picture of the deformation. The scientific advances will be applied to warm forming of two high strength alloys with different microstructures, namely AA6022-T4 and AA7050-T6.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

This project studies the role of glycogen synthase kinase-3 beta (GSK3B) in modulation of ethanol consumption. GSK3B has been implicated as having an important role in synaptic plasticity and learning. Additionally, GSK3B has been suggested to modulate behaviors for other drugs of abuse. GSK3B has been shown by multiple studies to be inhibited by phosphorylation on residue Ser9 in the medial prefrontal cortex (mPFC) and nucleus accumbens (NAc) of rodents after acute ethanol exposure. However, we recently found that prolonged ethanol consumption causes habituation of this inhibition of GSK3B by acute ethanol in PFC. Our recent studies on rodent drinking behavior show that pharmacological or genetic targeting inhibition of GSK3B in forebrain Camk2a+ neurons will decrease ethanol consumption. However, the mechanisms of GSK3B modulation of ethanol consumption are unknown. This proposal will perform a detailed analysis of ethanol regulation of GSK3B activity, and the specific cellular and neural circuits involved in GSK3B modulation of ethanol consumption. Aim 1 will investigate cellular sites of acute ethanol-induced GSK3B phosphorylation (inhibition) in mPFC and study the time course, duration and mechanisms of chronic ethanol-induced habituation of the acute response to ethanol. Viral vector and Cre-LoxP genetic targeting will be used in Aim 2 to identify whether Camk2a-positive neurons in mPFC are the critical site for GSK3B modulation of ethanol behaviors, and will identify downstream circuits of these neurons. RNAseq studies and bioinformatics in Aim 3 will then study genomic responses downstream of GSK3B following selective deletion or over-expression, thus identifying critical gene networks functioning in GSK3B modulation of ethanol consumption. Finally, Aim 4 will investigate the long-term efficacy and potential end-organ toxicity of Tideglusib, a highly specific inhibitor of GSK3B shown in preliminary studies to decrease ethanol consumption in rodent models. Tideglusib is already approved for phase II clinical trials on disorders such as autism. Together, these studies are highly significant and novel, and will provide needed knowledge regarding the mechanisms of GSK3B modulation of ethanol consumption. This work may also implicate a new agent, tideglusib, for clinical studies on treatment of AUD.

Evidence of significant co-occurrence between pain and alcohol consumption are emerging. There is also indication that alcohol can induce acute analgesia with cross- sectional evidence that some individuals may be motivated to use alcohol to cope with pain. However, potential moderators and mechanisms of action remain largely uncharacterized and poorly understood. This proposal will focus on examining potential pharmacological and genetic mechanisms mediating the alcohol-pain connection using the mouse BXD recombinant inbred (RI) panel. The primary objective of this proposal is to identify novel genetic factors that contribute to alcohol acute analgesic effects and development of tolerance in mice. We observed for the first time strain differences between C57BL6/J and DBA/2J for alcohol-induced antinociceptive effects in the hot- plate test after oral administration. In Aim 1, we will examine and characterize alcohol?s analgesic properties in acute pain models after oral dosing in the mouse. In Aim 2, we will use BXD RI lines to map genomic regions, or QTLs, that are causally associated with susceptibility versus resilience to alcohol effects and the development of tolerance in the hot-plate test. In Aim 3, we will identify changes in the transcriptome associated with acute analgesic phenotype and tolerance of alcohol. We will measure changes in gene expression in relevant neuronal tissues (amygdala, periaqueductal grey and prefrontal cortex) associated with alcohol initial sensitivity and tolerance in extreme RI strains. In Aim 4, we will validate candidate quantitative trait genes and functional variants identified and ranked by Aims 2-3.

Project Summary ? Overall Alcohol use disorders (AUDs) represent a major public health burden. Genetic risk factors contribute critically to susceptibility to AUDs and are likely a result of many variants each contributing modestly to risk. Genetic studies in animal models and humans have to date made slow progress in identifying individual genetic risk variants. However, modern high-throughput approaches such as genome-wide association studies or genomic expression profiling promise to rapidly increase the pool of potential candidate genes influencing AUDs. This proposal for a P50 Alcohol Research Center presents a novel and highly integrated overall design to focus on both gene discovery and functional interpretation for the genetics of AUDs. This application is a renewal of our currently funded P50 that supports the VCU Alcohol Research Center (VCU-ARC), which was first funded with a P20 Developmental Center grant in 2009. We have made significant progress over the past 4.5 years and here seek to both continue aspects of our prior directions but also to extend our work into new areas. Our approach continues to be innovative and significant due to three novel features: 1) A focus on gene networks contributing to AUD-related phenotypes and ethanol behaviors, rather than single genes; 2) A cross- species genetic and genomics analysis to validate candidate genes and networks affecting ethanol behaviors; and 3) A highly integrative Center design with rapid data sharing across projects through a cross-species analysis pipeline to provide ranked gene lists or networks for further experimental validation in the component projects. We request five years of support for five research projects and pilot grants for genetic studies in worms, flies, mice, rats and humans. Three projects will represent new areas of study, while two others will renew their overall strategy of current projects but with novel areas of investigation. Two projects will be in human genetics with state-of-the-art statistical approaches to leverage the power of large genome-wide association and exome sequencing studies on phenotypes significantly associated with AUDs. All projects will be supported by an Administrative Core, a Bioinformatics and Analysis Core and a Rodent Behavioral Core. The scientific work proposed in these projects and cores is clearly greater than the sum of its parts, due to the highly interactive structure of the VCU-ARC components. The VCU-ARC is well positioned to become a national resource, making major contributions to the advancement of our understanding of the etiology of AUDs and subsequently their prevention and treatment.

Project Summary ? Pilot Projects The goals of this Pilot Project Core of the VCU Alcohol Research Center are to: 1) manage the progress and quality control for our two designated initial pilot projects, 2) solicit and evaluate future pilot grant proposals to further expand the scientific breadth of VCU-ARC, 3) encourage pilot grant awardees to submit proposals for independent funding and 4) support training opportunities for students and postdoctoral fellows within laboratories of funded pilot projects. Drs. Bjork and Edwards are experts in motivational and internalizing factors that influence risk for human alcohol use disorders (AUDs) and will be the Lead and co-investigator of Pilot Project 1, entitled ?The role of neurocognition in polygenic risk for alcohol use disorder and comorbidities?. They have proposed a novel study to integrate data on the neurocognitive performance of individuals with severe AUD and uncover the genetic relationships leading to AUD risk. Their pilot will interface with the longitudinal genetic studies proposed in Projects 4 and 5 (Dick and Webb) and may inform analyses in Core 3 (Bacanu). Furthermore, if successful, Pilot Project 1's work will provide solid preliminary data for future funding applications assessing the neurocognitive and genetic contributions to risk for AUD. The second targeted pilot will be conducted by Dr. Joel Schlosburg, an expert neuropharmacologist with expertise in drug and alcohol use disorders, and is entitled Dissection of the acute and dependent responses to ethanol in rats lacking fatty acid amide hydrolase?. Pilot Project 2 will use a newly established FAAH knockout rat on an outbred background to test the role of FAAH on ethanol sensitivity, intake and preference. This pilot will expand the knowledge of the impact of potential future FAAH inhibitor development for use in clinical trials, with at least three separate such inhibitors in various phases and endpoints. These studies may also be the first to demonstrate whether NAPE-PLD is the primary synthetic enzyme supplying fatty acid amide substrates to FAAH. We will solicit other proposals in year 2 and fund the two best of them in years 3-4, then solicit proposals again in year 4 to fund one in year 5. The pilot program is an important component of the VCU-ARC that will enable us to attract new and innovative researchers to interact with Center investigators and expand high quality alcohol research at VCU.

Project Summary ? Administrative Core The Administrative Core of the Virginia Commonwealth University Alcohol Research Center (VCU-ARC) will provide the centralized administrative functions of the VCU-ARC. This entails supervision and coordination of five research projects (Projects 1-5), and three additional cores focusing on our pilot project program, our Rodent Behavioral Core and our Bioinformatics and Analysis Core. Additional responsibilities will include outreach and teaching within VCU and dissemination outside the University of accomplishments of the alcohol research program of the VCU-ARC. Overall scientific and administrative oversight will be conducted by the Center and Scientific Directors (Drs. Miles and Kendler), with assistance from the Steering Committee, who will be responsible for monitoring scientific productivity and integrity of the Center as well as the general scientific research direction of the Center. A Program Advisory Committee will assist the Directors and Steering Committee in administrative oversight of the Center and selection of pilot projects for years 3 through 5. The Administrative Core will assist active collaboration and communication between all Center components through a series of biweekly meetings consisting of research presentations, literature reviews or seminars by invited distinguished scientists in the alcohol research field. These meetings and invited seminars will be open to the VCU research community to encourage expansion of alcohol research at VCU. The Administrative Core will also oversee functioning of the two research cores to foster their active use by Center investigators. Ultimately, it will be the goal of the Administrative Core to oversee development of the Center as a national resource and to continue expanding the scope and quality of alcohol research at VCU.

Project Summary ? Project 1 Over the last four years of P50 funding to the VCU Alcohol Research Center (VCU ARC), our laboratory used Diversity Outbred (DO) mouse mice from Jackson Laboratories (http://do.jax.org) for behavioral genetics and initial genomic studies on progressive ethanol consumption. DO mice originate from 8 progenitor mouse strains chosen to maximize genetic diversity and utilize a breeding scheme producing a high degree of heterozygosity for fine mapping complex traits such as ethanol consumption. As such, DO mice more faithfully mimic genetic aspects seen with alcohol use disorder (AUD). As expected, based upon preliminary work in DO progenitor strains, behavioral studies using a progressive ethanol consumption model (intermittent ethanol access, IEA) on over 600 DO mice showed a broad distribution of consumption values (~0.5?38 g/kg/24h) that shifted to significantly higher intake over the 4 week experiment. Genotyping identified 10 genome-wide significant or suggestive QTL with LOD ? 6 and support intervals generally < 2 Mb. Strikingly, quantitative trait loci (QTL) for the first week of drinking differed from those during the last week of consumption. Ongoing haplotype analysis and integration of RNAseq data from prefrontal cortex have identified provisional candidate genes, including two that have been implicated in human genome-wide association studies on alcohol consumption or dependence. We hypothesize in this renewal that extension of this DO behavioral QTL and genomic data will identify novel candidate genes and gene networks contributing to ethanol consumption behaviors in mice and that such data will inform existing and future human genetic studies and therapeutic efforts on AUD. Our specific aims thus describe: 1) Further expression genetics analysis in nucleus accumbens, allele specific expression analysis, and chromatin 3D conformation analyses to identify and refine a list of positional candidate genes for behavioral QTL associated with initial ethanol intake vs. progressive consumption; 2) Gene network analysis of RNAseq data in both prefrontal cortex and nucleus accumbens across 200 DO mice to identify networks and possible mechanisms tightly associated with consumption differing between first and last week; and 3) Validation of candidate genes or network hubs as functioning in ethanol behaviors, including initial or progressive ethanol consumption, using invertebrate models (collaboration with Projects 2 and 3 of this Center proposal), rodent models (this project and Rodent Behavioral Core). Further, the work of this project will inform and be informed by novel human genetic studies described in Projects 4 and 5. Throughout this project, the VCU ARC Bioinformatics and Analysis Core will provide critical support for analysis of RNAseq data and Capture-C chromatin conformation studies, and the choice of candidate genes and networks. This novel gene discovery and network analysis effort will have major interactions with all other components of VCU ARC and will inform the field of alcohol research with understanding of mechanisms involved in the transition to abusive drinking, and candidate genes/mechanisms for potential targeting by future therapeutic efforts.