Core A: Administration, Human Resources, and Data Analysis
Core Leader: Akira Sawa, M.D., Ph.D.
The Core A (Administrative Core) is a large core that comprises sub-groups of administration, distinct resources of human materials, and data analyses/database construction. For efficient and central operation of the overall center, we have integrated these distinct components under the leadership of Core A PI, who also serves as the center director. This Core has three major aims: administrative leadership, organization of human bio-resources, and data organization/database maintenance.
In Aim 1, this Core will lead financial administration, organization of human and animal subject protocols, and coordination of research meetings and annual retreat inside this Center. This Core will provide scientific leadership to the entire Center by centrally organizing scientific issues towards uniform conclusions inside the Center, and coordinating collaborative research and material exchange with investigators outside the Center. This Core will maintain the web page of the P50 Conte center, organize education and outreach plans. Lastly, this Core will lead the Center in research rigor and integrity and make sure the members to observe data/resource sharing policies.
In Aim 2, this Core will organize and provide the center investigators with several types of established human resources, including three case-control sample sets for genetic study, the datasets of RNA-sequencing from human brains, blood from two prospective cohort samples, and human induced pluripotent stem cells. In Aim 3, this Core will continue development of a database in which the multiple types of data obtained in Projects will be systematically stored to support investigators for data sharing, analysis, and preservation. This is also a basis of data and resource sharing within the scientific community as well as public outreach. In summary, believe that this Core will continuously play a key role by providing the central service in administrative coordination, organization of human bio-resources, and data management/database maintenance. This core will lead research rigor and integrity on behalf of the Center. We will emphasize educational and public outreach plans, according to the successful protocol that we have built in the current P50.
Core B: Behavior
Core Leader: Jeffrey Leek, PhD, and Gianluca Ursini (Lieber Institute for Brain Development)
Our main goal is to identify the molecular expression changes induced by mutations in microtubule-associated genetic risk factors and stress exposure in human cell and mice models, which are developmentally regulated in human brains and differentially expressed in psychiatric disease conditions, such as schizophrenia (SZ). We hypothesize that mutations in microtubule-associated genes induce alterations in specific molecular pathways that are exacerbated by stress exposure in neuronal cells derived from human iPS cells and mouse models. We also hypothesize that these changes can be translated in a context relevant for human brain development and for psychiatric disorders, such as SZ. In order to test this hypothesis, we propose the following specific aims:
Aim 1 will determine the molecular expression changes in the homeostasis of induced neuronal cells as a consequence CRISPR/Cas9 mediated mutagenesis of KCTD13, DPYSL2, SDCCAG8 and CKAP5 by conducting a standardized RNA-seq analysis pipeline at base resolution and at transcript level. To quantity homeostatic change, we will measure the transcriptome, the primary cellular response, with and without exposure to dexamethasone as a model of stress.
Aim 2 will determine the molecular expression changes in the prefrontal cortex (PFC) of mice with mutations in Pcm1, 16pDup, 16pDup/Kctd13+/-, and Dpysl2 by conducting a standardized RNA-seq analysis pipeline at base resolution and at transcript level. We will also examine if observed changes are exacerbated by adolescent social isolation stress.
Aim 3 will address the question whether our findings in Aim 1 and 2 can be translated in a context relevant for brain development and SZ, by comparison of our RNA-seq data and two unique human post-mortem datasets, available in Core A.
Our gene expression and a network/pathway analysis as well as molecular profile comparison will detect specific molecular alterations associated with microtubule-associated genetic variants and environmental exposures in human cells and animal models, which may contribute to understand the molecular landscape underlying the brain development and the pathophysiology of psychiatric conditions, such as SZ.
Core C: Behavior
Core Leader: Michela Gallagher, Ph.D.
Research under the current proposal is designed to determine behavioral alterations relevant to schizophrenia (SZ), which is elicited by activation of stress-associated cascades and the E-I imbalance in the prefrontal cortex in mouse models that carry microtubule-associated genetic variants. We will also study how adolescent social isolation exacerbates these changes at the molecular, circuitry, and behavioral levels in collaboration with three Projects. In addition to performing basic behavioral characterization of the mouse models, this Core will conduct behavioral assessments for neurocognitive domains that are mediated by medial prefrontal cortex and orbitofrontal cortex, such as behavioral flexibility, including outcome expectancy in goal directed behavior, and working memory. Our rationale for use of specific behavioral assessments aimed at prefrontal systems is grounded in research that
1) documents the critical role of prefrontal cortex (PFC) circuitry in the information encoding mechanisms required to support behavioral performance in these paradigms across rodent and primate species,
2) implicates PFC systems supporting these functions in SZ by anatomical and physiological/neuroimaging evidence from patients,
3) demonstrates a vulnerability of such PFC-mediated behaviors/networks to stress exposure, including underlying mechanisms in PFC. Importantly, the assessments hold potential for the study of species-conserved cognitive mechanisms in the human brain to provide translational opportunities based on this preclinical research program.
As such, the work in Core C is consistent with the Research Domain Criteria (RDoC) approach for advancing a biological understanding of the pathophysiology of major psychiatric illness and creating a platform for discovery of new therapeutics. Core C will consult with the investigators of the Projects both for conducting basic behavioral assays, for implementing behavioral experiments with a focus on analytically powerful protocols targeting PFC-mediated behaviors, and will lead data analysis and interpretation of findings in behavioral studies under the research program. In addition to its scientific value to the immediate objectives of the research program, the work of this Core may have broader translational significance in the neuropsychiatric field. As the molecular pathways under investigation are better understood in the context of behavioral profiling, the work of the core could identify assessments that are best suited to target dysfunctional mechanisms in animal models of SZ. In this way the proposed behavioral research may yield innovative findings of more general preclinical importance.