Jin Lee, D.Phil - US grants

The role of the Histology Laboratory Core is to coordinate and provide professional and technical services for proper handling and processing of histopathologic materials. Specific aims are as follows: 1) to prepare routine hematoxylin and eosin (H & E)-stained pathology slides for histopathologic examination of the bronchial biopsy samples, and 2) to prepare and distribute tissue sections required for special histopathologic examinations and in-situ hybridization studies. The main purpose of this project is to develop effective chemopreventive approaches in former smokers and to identify potentially useful biomarkers of intermediate endpoints. We expect to deal with a large bulk of histopathologic materials in this project. In addition, some studies require special care in preparing the tissue sections. For example, in- situ mRNA hybridization studies (e.g., retinoic acid rector gene expression), the slides should be prepared using a specially treated water to avoid RNA degradation, which can not be accommodated by the regular pathology department personnel. Therefore, it is essential to establish a separate core facility for successful control of this project. As the clinical trial progresses to attain the targeted sample size, more Core effort will be required in the preparation and distribution of tissue sections for the translational research projects. The role of this Core will become increasingly important because proposed studies and further progress in the field of carcinogenesis research may identify new cellular, biochemical, or molecular genetic markers of interest. These will need to be tested and expanded using the tissue samples obtained through this project.

This program project will require the procurement and processing of a large bulk of histopathologic materials. These materials need to be processed, not only for routine histopathologic diagnosis of the tumor and premalignant lesions, but also for the various immunohistochemical and in situ hybridization studies proposed in the basic research projects. Preparation of tissue samples for these studies cannot be processed by a regular pathology laboratory since they require special care in their handling. For example, to obtain optimum results with mRNA in situ hybridization, all glassware should be baked, and tissue sections need to be handled with special care to avoid RNAase contamination. This Histology Laboratory Core will respond to such a special need and will prepare and distribute adequate tissue samples for the studies proposed in Projects 4 through 6. It will also function as a liaison with the Department of Pathology to coordinate the handling of pathology slides and paraffin blocks sent from collaborating institutions (e.g.,RTOG and CCOP institutions for Project 1). In addition, this core will process the immunohistochemical staining for the conduct of Project 3. Another major function of the laboratory will be the procurement and storage of both normal and malignant tumor samples for future use. This Core facility will become increasingly important since the proposed studies may find new molecular biologic of biochemical markers of interest that would warrant re-examination of some samples in the future.

[unreadable] DESCRIPTION (provided by applicant): The goal of this project is to understand how repeated experience can stably modify a neuron's activity by studying the molecules that affect the nuclear localization of the cGMP-dependent protein kinase EGL-4 during long-term odorant adaptation in C. elegans. This understanding will provide us with insight into mechanisms of learning and memory and molecular processes that underlie addictions. Nuclear localization of EGL-4 in odorant sensory neurons is necessary for the transition from short to long-term adaptation that occurs upon continual stimulation to an odorant. We propose to study the molecules that regulate this nuclear translocation. By utilizing a GFP-tagged EGL-4 molecule to visualize in real-time the behavior of EGL-4, we will examine the localization of EGL-4 in known components of both odorant chemotaxis and adaptation. We will also genetically screen for individuals that are defective in EGL-4 nuclear translocation to identify new components of adaptation that regulate the transition to long-term adaptation. The results from this study will give us a clearer understanding of neuronal plasticity. [unreadable] [unreadable]

A key in understanding the etiology and interventional outcome of neuropsychiatric diseases is the ability to analyze the brain's functional circuitry through precisely controlled stimulation mechanism, while at the same time non-invasively monitoring changes in neuronal activity. The use of conventional stimulation electrodes is constrained by the lack of its ability to selectively target different neuronal populations. Likewise, electrode-based neural stimulation does not necessarily mimic endogenous neural activity, and a spatial propagation ofthe activity in a biologically realistic fashion cannot always be guaranteed. The monitoring of the resulting brain activity on the other hand includes the use of recording electrodes that wili provide high temporal resolution measurements. However, the lack of "anatomical awareness" of recording electrodes is a limiting factor for the analysis of functional circuitry that involves multiple, and possibly elusive brain areas. Blood oxygenation level dependent (BOLD) functional MRI (fMRI) with Its non-Invasive, whole-brain coverage capability is promising for such large-scale neuronal monitoring. But current fMRI schemes struggle with problems of image distortions and lack of sufficient spatial resolution. The candidate of this K99, Pathway to Independence grant is a superbly trained MR scientist now seeking to bridge the gap between fMRI monitoring and targeted neural stimulation schemes that exist today. In this proposal, the candidate proposes a coordinated development of a highly innovative molecularly targeted neuro-optical stimulation method with a de novo distortion-free, high-resolution functional MRI technique the candidate has developed in recent years. With this new method, specific types of neurons can be molecularly targeted for interrogation, endogenous neuronal activation elicited, and the resulting pattern of neuronal activity monitored at an exceedingly high spatial resolution without distortions. This new capability to non-invasively monitor brain activity at high spatial resolution, while controlling the neuronal activity with high functional precision, will provide a powerful future tool for studying the mechanisms of neuropsychiatric diseases. This will lead to better understanding ofthe disease mechanism as well as the development of new treatments.

DESCRIPTION (Provided by the applicant) Abstract: This proposal suggests a consilience of bold new concepts in genetics, stem cell biology, and functional magnetic resonance imaging (fMRI) to address a singular question that could never be tapped previously: can we directly map the functionality of stem cell-driven neural circuit regeneration in vivo? Despite the debilitating character of central nervous system (CNS) diseases, and the urgency for therapeutic development, the complex nature of the neural fabric underlying CNS diseases such as spinal cord injuries, Parkinson's disease, Alzheimer's disease, multiple sclerosis, and stroke, substantially negate their viable cure to date. Here, the fundamental ability of stem cells to regenerate non-dividing cells, as well as recent development of induced pluripotent stem cells (IPSC) gives fresh impetus for a whole new class of innovative treatments where damaged neural circuitry might be partially or fully restored by stem-cell induced neurogenesis. We seek to be instrumental in this pivotal endeavor by introducing a completely novel way of directly assessing the functionality of stem cell driven neural circuitry in vivo. This will be achieved by combining genetic techniques that will introduce modulatory and/or reporting capability to neural cells based on its cell type. We will strategically introduce viral vectors to both the underlying neural circuit as well as transplanted stem cells. For stem cell transfection, we expect our proposed technique to allow modulatory/reporting capability from only those developing into specific cell types. This cell-specific modulation/reporting capability will then combined with a novel distortion-free fMRI imaging technique, permitting non-invasive and high-resolution visualization of the regeneration processes. This project, upon its success, will provide direct functional assessment capabilities for the regenerated nerve tissue in vivo. This in turn will provide key guidance for developing novel stem cell therapies for CNS diseases. Public Health Relevance: The complexity and functional nature of neural circuitry makes central nervous systems (CNS) diseases such as spinal cord injuries, Parkinson's disease, Alzheimer's disease, multiple sclerosis, and stroke particularly challenging for therapeutic approaches. While recent development of induced pluripotent stem cells (IPSC) gives fresh impetus for a whole new class of innovative treatment landscapes for patients with debilitating CNS diseases, the ultimate functionality of stem cell induced CNS regeneration remains elusive. This project, upon its success, will provide direct and functional assessment capabilities for the regenerated nerve tissue in vivo. This in turn will provide key guidance for developing novel stem cell therapies for CNS diseases.

1056008
Lee

This proposal utilizes a brand new technology called optogenetic fMRI (ofMRI)(pioneered by Dr. Lee) combined with advanced imaging, computation, and applied mathematical algorithms. Optogenetics, is a genetic technology that allows neural circuit elements to be triggered selectively based on their genetic identity with temporal precision, while passband b-SSFP fMRI, provides high quality functional brain activation maps to be obtained of MRI is a technology that combines the two to visualize precise causal response of the brain circuit upon selective triggering. This allows the brain circuit element's causal role in the system level outcome to be evaluated. However, in order to parse through the massive combination of the brain circuit's response, real-time interactive monitoring is necessary while the small, dense structures of the brain requires high resolution acquisition. The PI aims to design and implement novel parallel computation and compressed sensing (CS) reconstruction algorithms to achieve these goals. The implementation will then be utilized to look at brain circuit responses to a range of frequency stimulation. In particular, the motor cortex (M1), striatum, thalamus, and substantia nigra (Stn) will be stimulated in order to precisely map the system known to be correlated with the motor symptoms of Parkinson's disease. Brain activity amplitude, shape, and distribution will then be carefully compared to analyze the circuitry.

DESCRIPTION (provided by applicant): Children from low-income language minority backgrounds begin kindergarten at a significant disadvantage compared to their English-speaking peers, highlighting the need to provide them with enriching educational experiences in early childhood. Many state readiness standards now highlight preschool science as a key domain in the preparation of young children for the transition into formal schooling (Head Start, 2007). Yet, the lack of studies with rigorous research designs to evaluate the effectiveness of preschool science curricula has prevented researchers from drawing conclusions about best practices. In addition, preschool science curricula have been designed for classrooms made up of European American children from middle income backgrounds and the effectiveness of these programs have not been tested with low-income Latino preschool children from Spanish- speaking backgrounds. The overall goal of the proposed study is to design and test the efficacy of a preschool science curriculum for low-income Latino children that focuses on improving their conceptual understanding of germ contagion and contamination, and food and nutrition, an area of interest to NIH related to developing creative and innovate research education to deliver information about healthy living in science to children. Our study is novel in that it integrates health and biology concepts in a multi-unit science curriculum, instead of introducing health information as a stand-alone topic outside of science, typical of preschool programs. In the proposed research, 40 preschool classrooms will be assigned randomly to one of two experimental groups: 1) a treatment group that receives the biology-based health science curriculum; 2) an attention control group that receives a standard health curriculum from published, on-line materials. Within each experimental group, half of the classrooms will receive the curriculum in Spanish and the other half will receive the curriculum in English. All children, ages 4 and 5, will participate in pre- and postest assessment sessions. We expect that relative to control group, children will show an increase in conceptual understanding of health concepts related to biological process, and science inquiry skills as measured by their capacity to ask questions and generate explanations. There will be significant increases on measures of science understanding and inquiry skills for both groups of children (those receiving the instruction in Spanish and those in English) although the overall effects of the experimental curriculum will be stronger for children receiving the instruction in their primary language (Spanish).This study will provide vital information for the development and dissemination of a biology-based preschool health science program particularly for low-income Latino children from Spanish-speaking backgrounds, but appropriate for different types of learners.

DESCRIPTION (provided by applicant): An important challenge in drug development for Alzheimer's Disease (AD), as well as many other neurological diseases, is the accurate pre-clinical predictability of the future clinical efficacy and side effects of the compound. The complexity of the neurological system makes it extremely difficult to infer impact of the candidate compound using surrogate in vitro information, ex vivo data, or simple animal behavior models. Our proposal aims to fundamentally transform this process through accurate disease circuit characterization along with in vivo full brain quantification of the drug administration impact longitudinally over time in the same animals utilizing the optogenetic functional magnetic resonance imaging (ofMRI) technology. ofMRI is a novel technology combining cell type specific, temporally precise optogenetic control with fMRI readouts. The first proof of concept study published by the PI demonstrated ofMRI technology's ability to accurately visualize brain-wide neural activity resulting from cell types specified by its genetic identity, cell body location, and axonal projection target. With further developments in ofMRI technology to achieve high-throughput, high spatial resolution, and awake scanning, the PI's lab has preliminary data showing the ability to characterize whole brain network response specifically associated with basal forebrain cholinergic neurons. Selective stimulation of cholinergic neurons in the basal forebrain with defined temporal patterns was shown to elicit widespread neural activity including the hippocampus and neocortex. Such direct functional visualization of the network activity with the ability to longitudinally track in vivo can be used o characterize the disease progress, actively identify novel target circuits, and also monitor how different doses and course of treatment impacts each individual animal over time with disease progress. This is in contrast with conventional ex vivo technology that relies on sacrificing a large number of animals at different time points, which increases variance, cost, and time while only obtaining passive, surrogate markers of function or difficult to reproduce behavioral tests. I this proposal, we will quantify network function across normal, and mutant APP mice to quantify network function changes associated with cholinergic neurons of the basal forebrain. With these quantifications, we aim to establish this approach as a new platform for quantitative drug design and evaluation. With the ability to observe network engagement associated with key neural circuit elements implicated in AD, we will identify the AD circuit mechanism and also test a candidate drug that has already shown good electrophysiological, behavioral, and anatomical efficacy. We aim to demonstrate ofMRI-based observation of therapeutic effects. The prevention and/or reversal of AD-related circuit impairment could be fully or partially effective and we will e able to directly observe locations of the changes associated with specific neuronal populations in live animal brains during disease progression and drug therapy. Resting state fMRI studies will also be conducted in parallel to increase translational potential into clinical settings.

DESCRIPTION (provided by applicant): Central thalamic deep brain stimulation (CT-DBS) is a promising therapy for restoring consciousness in patients in coma and vegetative state by changing the arousal state. Early experimental studies have established a causal link between central thalamus electrical brain stimulation and forebrain arousal. Clinical investigators in the 1960s and 1970s considered the potential relevance of the findings as a method for restoration of arousal and consciousness in chronically unconscious patients and carried out pilot case studies of electrical stimulation. However, despite eye opening and autonomic signs consistent with arousal effects, no reports described sustained recovery of interactive behavior. Following on these early case reports, a multicenter study involving a total of 49 patients was carried out. Deep brain stimulation resulted in increases in arousal and associated physiological responses in the majority of these patients but there were unfortunately no changes in behavioral responsiveness. More recently, a single subject study provided the first compelling evidence that some severely brain injured patients in minimally conscious state (MCS) may benefit from CT/DBS. The overall findings indicated significantly improved behavioral responsiveness with a combination of immediate as well as slowly accumulating, though long-lasting effects. However, this type of response has only been observed in a single subject and has been difficult to reproduce. While these earlier findings raise the possibility that using CT/DBS to improve consciousness in severe traumatic brain injury could be efficacious, many critical challenges lay ahead. These include defining the mechanisms of action and optimizing stimulation targets and parameters to make CT/DBS a reliable clinical treatment. In this proposal, we aim to overcome these challenges. We will develop and utilize a novel optogenetic functional magnetic resonance imaging approach that will enable us to systematically understand the underlying mechanism of action of the CT/DBS therapy with unparalleled clarity. Elucidating the mechanism of CT-DBS therapy will allow us to optimize the stimulation target, stimulation parameters, and even help stratify patients for inclusion into such therapeutic modalities. In Preliminary Studies we have established the effectiveness of the ofMRI approach at identifying key locations of stimulation and, importantly, function associated with CT-DBS. We will first conduct ofMRI, EEG, and behavioral studies in normal animals to define circuit mechanisms involved in both acute and long-term stimulation. Then, we will evaluate the optimized parameters of stimulation and their role in restoring consciousness in animal models of TBI. Knowledge of CT-DBS mechanism and optimization parameters will be invaluable for future clinical trials, understanding of mechanisms of arousal, consciousness, and neuromodulation therapy.

This project will implement a Robert Noyce Teacher Scholarship Phase II project, STEM Teachers for English Language Learners: Excellence and Retention (STELLER), at the University of California, Santa Barbara (UCSB). STELLER addresses the growing national need to recruit, prepare, and provide effective induction support for new STEM middle and high school teachers with degrees in STEM fields and strong content-specific pedagogical preparation. A unique aspect of this project is the intent to focus on preparing new teachers who can meet the needs of English Language Learners (ELLs), students whose difficulties in speaking, reading, writing, or understanding English may hinder their learning. This program seeks to recruit undergraduate STEM students across all disciplines for entry into a 13-month post baccalaureate program leading to certification and a career teaching in high needs secondary schools. A total of 40 scholars will be supported over the five years of the project. The new teacher scholars will obtain teaching positions in high-need secondary schools with a particular emphasis on placement in the Santa Barbara Unified School district but also elsewhere on the California Central Coast in Santa Barbara, Ventura, San Luis Obispo, and Kern counties.

STELLER's team will consist of UCSB faculty from science, mathematics, engineering, computer science, and education; teachers and administrators from the high-need Santa Barbara Unified School District (SBUSD); directors from UCSB's Office of Education Partnerships; and community leaders from the Santa Barbara County K-20 STEM Council. The program will continue efforts begun under the Noyce Phase I CalTeach at Santa Barbara (CTSB) to increase the quality, number, and diversity of secondary science and mathematics teachers who graduate from UCSB's post-baccalaureate Teacher Education Program (TEP). Starting at the undergraduate level, a focus on English Language Learners (ELLs) will be added so as to thoroughly prepare secondary science and mathematics teacher candidates to teach the disciplinary-specific language, core ideas, and practices outlined in the Next Generation Science Standards and the Common Core State Standards in Mathematics. The trajectories of CTSB and STELLER Teacher Scholars will be tracked to generate new insights into effective teacher education. This project will produce a 24% increase in UCSB's production of science and mathematics teachers. Evaluation efforts will include an assessment of activities specific to this project as well as continued assessment of the success of beginning teachers from the initial Phase I Noyce Program at UCSB. A mixed methods approach will be used in project evaluation. Data regarding the program operation will be collected through surveys and interviews with all participants. Data regarding teacher effectiveness and training will be collected in classroom observations, tracking of average state standardized test scores of students, and performance on the state-mandated teaching performance assessment.

? DESCRIPTION (provided by applicant): Recent success of neurostimulation therapies such as deep brain stimulation (DBS) for Parkinson's disease (PD) support the importance of understanding how activity of specific local neuronal population influence the overall brain network to drive behaviors such as reversing tremors in Parkinson's disease. Given the resulting complex behavioral output, it is likely that the effects of brain stimulations are not limited to simply changing local neuronal activity. The local change is driving neural activity in many regions of the brain to give rise to the therapeutic effects. However, this important neurobiological question of how large-scale network activity relates to behavior still remains largely elusive. Understanding of how specific neuronal population functionally relates to the overall brain enables us to systematically design therapeutics for neurological diseases based on our concrete knowledge of the circuit function underlying behavior. The main therapeutic goal for neurological diseases lies in reversing the behavioral phenotype such as essential tremors, which are a direct consequence of loss of proper circuit function. If the circuit function underlying behavior can be directly visualized, the potential for therapeutic intervention is limitless. Therefore, in this proposal, we aim to start reverse- engineering global brain dynamics associated with the basal ganglia circuit and to understand how they relate to motor behavior. The novel optogenetic functional magnetic resonance imaging (ofMRI) technology, enables us to selectively trigger specific neuronal populations within the brain while monitoring how activity in regions across the brain are altered as a result of such stimulations. Optogenetics enables cell-type specific, millisecond-scale, activity modulation using light while high-field fMRI tracks resulting responses in live subjects across the whole brain. In the initial study, it was shown tha specific cell-type triggered fMRI responses could be measured throughout the brain with temporal precision. Since we first developed the ofMRI technology, we developed advanced imaging technologies to enable high-throughput, high-resolution images in live subjects. With these advances in place, we acquired preliminary ofMRI datasets, through which we have evidence that dopamine D1 and D2 receptor expressing medium spiny neuron (MSN)-driven dynamic interactions across the whole brain can be reliably measured across multiple synapses. Electrophysiological recordings also show strong evidence that the time course of the ofMRI signal closely matches underlying electrical activity patterns. With this unprecedented ability to obtain global brain dynamics associated with cell- type specific modulations, we aim to determine the global direct and indirect pathway functions. These measurements will then be computationally modeled to provide a mechanistic understanding. In addition, resting-state fMRI measurements will be made during systematically increased and decreased excitability of D1 or D2 MSN. This will enable us to evaluate how the direct and indirect pathway imbalance is reflected in resting-state fMRI measurements, and allow direct translation of the findings into clinical neuroimaging.

Project Summary / Abstract: The blood oxygenation level dependent (BOLD) functional Magnetic Resonance Imaging (fMRI) signal source has been long debated since the invention of fMRI in the early 90s. While fMRI is one of the most successful technologies utilized in numerous studies, the debate over the source of fMRI signal source continue to generate controversies over the utility of fMRI and the interpretation of fMRI studies. One important aspect in the study of fMRI signal source is that it is fundamentally difficult to study the fMRI signal source in isolation in one brain region with one cell type. fMRI signal represents an intricate interaction between the vascular system and the neural activity and it is bound to depend on cell type and vascular composition of each region of the brain. In this context, basal ganglia thalamo-cortical circuit provides a unique opportunity to understand the fMRI signal source. Many of the basal ganglia circuit elements contain largely inhibitory cell populations with remote projections. This enables the possibility of driving neural activity throughout the brain with distinct temporal dynamics, which can potentially help delineate fMRI signal with more specificity on each cell type?s role. In addition, the basal ganglia thalamo-cortical system is an important network in the brain that consists of multiple nodes throughout the brain forming a distributed network that real- time controls important behavior such as movement. It is one of the key networks that require large-scale imaging methods such as fMRI to understand its overall dynamics. Specifically, studying the fMRI signal source associated with this network function will directly enable us to apply the knowledge to understanding an important neurobiological question. Our experimental strategy consists of three main components. In the first aim, we will measure cell type specific whole brain network function during D1- or D2- MSN stimulation utilizing simultaneous ofMRI and electrophysiology. In the second aim, we will conduct optical imaging at locations identified by preliminary ofMRI studies to confirm sources of fMRI signal with cell type specificity at each location. Then, we will computationally model the measurements to systematically understand the source of the fMRI signal against frequency components of the electrophysiology signal and also cell types at each downstream location from the stimulation site. We will utilize this knowledge to then construct models of whole brain circuit function with both stimulation cell type and downstream locations cell type specificity These aims are designed to understand the fMRI signal source in the context of D1 and D2 MSN regulation of global brain function across multiple synapses. We will also demonstrate the power of such studies in enabling a comprehensive model that can describe the interactions among all regions involved with both stimulation cell type and downstream local regional cell type specificity. This study will provide an important groundwork for understanding fMRI signal sources and how it can enable cell type specific whole brain function investigation with unprecedented precision.

Deciphering how the brain works could have untold impacts on medicine, technology, commerce, and our understanding of ourselves. For example, advances in neurotechnology could lead to brain-machine interfaces to overcome sensory impairments and loss of movement due to neurodegenerative disease. Many of the most important advances in neuroscience have required interaction with technical fields such as physics, electrical and chemical engineering, bioengineering, statistics, and computer science, and this will increasingly be the case as the field advances. However, the path for top students from these disciplines to enter the field of neuroscience has always been challenging because they lack the appropriate background and awareness of key questions and technological limitations in the field. This National Science Foundation Research Traineeship (NRT) award to Stanford University will accelerate fundamental developments in neuroscience by attracting promising young talent from these technical disciplines to neuroscience and training them to be leaders in the field. The program will allow students to apply technological developments in diverse fields to the most important problems in neuroscience today and train a new generation of neuroscientists who will bring these technologies to fruition in academia, medicine, and the private sector. The project anticipates training thirty (30) PhD students, including twelve (12) funded trainees, from physics, electrical and chemical engineering, bioengineering, materials science, computer science, and other technical fields.

This traineeship program consists of a novel integrated curriculum of coursework, internship and training experiences, and outreach to achieve its goals. The program will emphasize training for acquiring and analyzing vast data sets, enabling an understanding of nervous system circuitry at a scale that was unimaginable just a few years ago, and connecting the novel data to Stanford's strength in theory, inference from large data sets, and computational modeling. The program will introduce a rigorous multi-year curriculum for trainees, building on their home-discipline training and allowing them to collaborate with each other and with the members of the Neurosciences PhD program. Training will leverage the highly successful Stanford ADVANCE program that supports new PhD students with a special summer program prior to the start of graduate training, and build on it with several approaches customized to this program. The program will be specifically designed to optimize trainee preparation for a career in academia or in a technology industry setting, utilizing internship placements with both startups and established corporations.

The NSF Research Traineeship (NRT) Program is designed to encourage the development and implementation of bold, new potentially transformative models for STEM graduate education training. The program is dedicated to effective training of STEM graduate students in high priority interdisciplinary research areas through comprehensive traineeship models that are innovative, evidence-based, and aligned with changing workforce and research needs.

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.

Project Summary / Abstract: One of the central goals of Neuroscience is to understand the brain circuit mechanisms underlying behavior. To understand the brain circuit mechanisms, it is becoming increasingly clear that the whole brain connectivity and function needs to be measured to bridge scales between micro- circuit mechanisms and whole brain system function. The requested Bruker BioSpec 70/40 USR has the state of the art large bore 7T magnet combine with high-performance gradients and multi-channel transmit-receive radiofrequency system for rapid, high-resolution imaging. The large bore system with features customized for state-of-the art Neuroscience studies will critically enable new technology development for the understanding of neural circuit mechanisms as well as enable increasing number of users with NIH funding that wish to utilize more MRI time. This advanced imaging system will be part of a core facility, located in the new Stanford Neuroscience Institute building. The requested Bruker BioSpec 70/40 USR imaging system will support NIH funded projects from 22 researchers and this number is expected to grow with a state-of-the art system that is located centrally in the neuroscience community at Stanford. The projects from the 22 researchers investigate a wide range of topics, including: Optogenetic fMRI and the investigation of global brain circuit mechanisms (Lee), Investigating post-stroke recovery mechanisms using imaging techniques (Steinberg), Developing Multivariate Dynamical Systems based Markov chain Monte Carlo (MDS-MCMC) algorithms (Menon), Using optogenetic control/modulation to study anhedonia (Deisseroth, Glover); Neurostimulation by Ultrasound: Physical, Biophysical and Neural Mechanisms (Butts-Pauly), Defining Cell Type Specific Contributions to fMRI Signals (Lin); Brain-wide consequences of optogenetic fMRI of LC in ion channel mutants (de Lecea), Imaging Global Brain Circuit Mechanisms Underlying Social Interactions (Shah), Imaging circuit mechanisms of closed- loop intervention in epilepsy (Soltesz), Studying the basal ganglia circuitry with electrophysiology and optical imaging and optogenetic fMRI (Ding), Innovating high-resolution novel imaging approaches to elucidate mechanisms of prion-like spreading of neurodegenerative disease (Gitler), Mechanogenetics (Liphardt), MRI/optogenetic mapping of epileptogenic circuits and effects of prophylactic agents following traumatic brain injury (Prince), Transforming neurostimulation methods for TMS (Etkin), and Establish guidelines for using dMRI fiber tracking to localize deep brain stimulation targets (McNab). These studies investigate critical functional and structural questions regarding fundamental neurobiological processes and cover NIH research areas with implications for diverse aspects of human health and disease, including Epilepsy, Parkinson?s and Alzheimer?s? disease, traumatic brain injury, stroke, depression, and autism. All projects will benefit significantly from simultaneous multi-modal neuromodulation combined with multi-modal recording/imaging; this combination of capabilities is most effectively provided by the requested Bruker BioSpec 70/40 USR system.

Project Summary / Abstract: Many neuroscience studies have shown that specific cell types within a brain network have unique contributions to behavioral output and that even a single neuron makes connections to large portions of the brain. Therefore, in order to truly get at the problem of uncovering brain function we need measurements with cellular specificity across the whole brain during behavior. As such, due to technological limitations, our current understanding of global brain circuit mechanisms is extremely limited. My recent development of optogenetic functional magnetic resonance imaging (ofMRI) technology provides a partial solution. However, challenges still remain: how do you non-invasively deliver cell type specific neuromodulation? How do you image the whole brain function in freely moving subjects? In this Pioneer Award proposal, I propose a novel approach that enables non-invasive, cell type specific, whole mammalian brain imaging in freely moving subjects. In particular, we propose to develop a non-invasive cell type specific stimulation in mammalian brain termed ?Mechanogenetics? and a functional ultrasound (fUS) imaging technology that can image whole brain function in awake-behaving animals. Mechanogenetics will utilize mechanosensitive ion channels expressed in selective cell types enabling neuromodulation using mechanical deflection from ultrasound probes delivered non-invasively instead of using optical probes that need to be surgically implanted. For imaging, miniaturized functional ultrasound technologies with high-resolution, 3D real-time imaging capability that can be mountable on the subject's head will be developed. The resulting ?Mechanogenetic functional ultrasound (MfUS)? technology will enable non-invasive flexible modulation of neuronal populations while the impact of such modulation can be monitored in freely moving animals across the whole brain with high spatiotemporal resolution. Instead of measuring large-scale neuronal activity associated with binary behavioral readout or complex behaviors related to single neuronal populations, my goal is to establish a new paradigm for understanding brain function, where cell type specific whole brain function during behavior can be monitored continuously. With such data, combined with computational modeling, whole brain algorithms of behavioral control can be constructed. Furthermore, the Mechanogenetics technology can bring cell type specific neuromodulation closer to human translation. Functional ultrasound technology development will also enable human brain function monitoring in non-laboratory settings. This will ultimately enable brain circuits to be engineered the way electrical engineers engineer electronic circuits allowing direct treatment of neurological disease including Alzheimer's disease and related dementias or directly manage pain addressing the opioid crisis.

An exciting new area in neurodegenerative disease research is the emerging phenomenon of prion-like spreading of neurodegenerative disease proteins. Prions are well established as the protein-based infectious agent underlying the spongioform encephalopathies. In these rare, albeit devastating, diseases the prion protein converts from the normal soluble form to the aggregated self-templating infectious form. This process initiates an inexorable spread of pathology and contingent neurodegeneration throughout the brain. But could this disease mechanism extend to the more common neurodegenerative diseases like Parkinson?s disease (PD) and Alzheimer?s disease and related dementia? If so, it represents a game changer in terms of understanding disease mechanisms and opens many new avenues for therapeutic development. However, any therapeutic or mechanistic investigation into prion-like spreading will require the development of powerful new imaging approaches to track the path of prion-like spread and understand how these protein aggregates alter brain function as they spread. We will need to understand why they take some routes but not others and how this impacts brain function. To address this challenge, we have formed an interdisciplinary team, consisting of an engineer and a geneticist. First, we will use state of the art brain clearing technology to obtain high-resolution images of prion-like protein propagation of the Parkinson?s disease protein ?-synuclein. We will monitor these aggregates as they spread from one neuron to the next, tracking their paths. These high-resolution brain wide 3D maps of alpha-synuclein spreading will empower us to identify gene expression patterns associated with spreading paths and to nominate genes for functional studies. Then, we will utilize advanced high-resolution optogenetic functional magnetic resonance imaging (ofMRI) to reveal the longitudinal effects of prion-like spreading on brain network activity and likewise the impact of neural activity on prion-like spreading. Our experiments will provide fundamental mechanistic insight into prion-like spread of neurodegenerative disease. The tools we apply and the lessons we learn will likely be broadly applicable to neurodegenerative diseases including Alzheimer?s disease and related dementias.

This project aims to serve the national interest by promoting engineering design that considers social, economic, and environmental factors. It will do so by supporting the distribution and adaptation of a Social Engagement Toolkit developed by the principal investigators. This resource provides instructors with teaching materials that help engineering undergraduates gain skills in considering social aspects of engineering problems when designing solutions and engaging clients and stakeholders. These skills align with the National Academy of Engineering?s 21st Century Challenges and are foundational for today?s engineers. However, these non-technical skills are often under-emphasized or absent from engineering curricula. The impact of the Social Engagement Toolkit on student learning will be examined across multiple courses and different institutions, including Georgia Institute of Technology, University of California, Fullerton, and University of Michigan, Dearborn. The results will inform how to adapt the Social Engagement Toolkit for different institutional contexts. A study of the instructors? attitudes to adoption of the Social Engagement Toolkit is also planned. The resulting adaptations of the Social Engagement Toolkit can produce an important, transferable resource to support socially engaged engineering education nationwide.

The Social Engagement Toolkit uses a hybrid learning block approach to provide on-demand virtual lessons combined with in-person coaching to help engineering students learn to integrate contextual information (e.g., physical, personal, social, cultural, and societal contexts) at every phase of their engineering work. The research component will assess the impact of the Social Engagement Toolkit on student learning and instructor adoption of the materials into their courses across institutional contexts and program settings. The research questions will be answered via a mixed methods study that includes surveys and focus group interviews. The impact of the Toolkit on student learning will be investigated via collection and analysis of pre- and post-assessment data. Both engineering instructors who adopt and those who do not adopt the Social Engagement Toolkit will be queried to understand barriers to adoption and to assist the adaptation of resources to suit instructors? needs. It is expected that the Social Engagement Toolkit will be a vehicle for promoting greater inclusiveness in engineering programs. The NSF IUSE: EHR Program supports research and development projects to improve the effectiveness of STEM education for all students. Through the Engaged Student Learning track, the program supports the creation, exploration, and implementation of promising practices and tools.

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.