Donna M. Ferriero - US grants

Support is requested for the study of regulation of the development of somatostatin-containing neurons in rat retina. A large quantity of somatostatin present early in the embryonic rat retina, with a significant decline in concentration prior to synapse formation, suggesting a role for the peptide in retinal synapse development. The first year of the proposed research will focus on the regulation of somatostatin synthesis at the transcriptional level in an effort to show that synthesis is turned off when somatostatin levels decline. Northern blot hybridization analysis will be utilized to establish the quantity of somatostatin mRNA expressed at key developmental timepoints. Then, in situ hybridization of the somatostatin message will be performed in tissue sections of developing retina to determine that the biosynthesis of somatostatin occurs in defined neuronal populations. To support the hypothesis that somatostatin binding sites will be studied in vitro and autoradiographically. Retinal cell suspensions will then be cultured to define more specifically the molecular mechanisms by which somatostatin is expressed. Firstly, mRNA will be measured and correlated with somatostatin content in this system. Then, the effects of cyclic AMP, which has been shown to regulate somatostatin mRNA accumulation in primary diencephalic cultures, will be monitored. These assays, employing forskolin stimulated activation of adenylate cyclase, will be performed to determine if cyclic AMP regulates somatostatin biosynthesis. Cysteamine, which reversibly depletes somatostatin immunoreactivity and biologic activity in the retina and central nervous system, will be added to tissue cultures to further study the interactions of cAMP activated synthesis in the depleted state. Finally, models of abnormal development including transgenic mice with obligate synthesis of somatostatin, and genetic strain of microophthalmic mice (mi/mi), will be used to study the patterns of somatostatin synthesis and expression in the abnormal state, both in vivo and in vitro.

Support is requested for the study of regulation of the development of somatostatin-containing neurons in rat retina. A large quantity of somatostatin present early in the embryonic rat retina, with a significant decline in concentration prior to synapse formation, suggesting a role for the peptide in retinal synapse development. The first year of the proposed research will focus on the regulation of somatostatin synthesis at the transcriptional level in an effort to show that synthesis is turned off when somatostatin levels decline. Northern blot hybridization analysis will be utilized to establish the quantity of somatostatin mRNA expressed at key developmental timepoints. Then, in situ hybridization of the somatostatin message will be performed in tissue sections of developing retina to determine that the biosynthesis of somatostatin occurs in defined neuronal populations. To support the hypothesis that somatostatin binding sites will be studied in vitro and autoradiographically. Retinal cell suspensions will then be cultured to define more specifically the molecular mechanisms by which somatostatin is expressed. Firstly, mRNA will be measured and correlated with somatostatin content in this system. Then, the effects of cyclic AMP, which has been shown to regulate somatostatin mRNA accumulation in primary diencephalic cultures, will be monitored. These assays, employing forskolin stimulated activation of adenylate cyclase, will be performed to determine if cyclic AMP regulates somatostatin biosynthesis. Cysteamine, which reversibly depletes somatostatin immunoreactivity and biologic activity in the retina and central nervous system, will be added to tissue cultures to further study the interactions of cAMP activated synthesis in the depleted state. Finally, models of abnormal development including transgenic mice with obligate synthesis of somatostatin, and genetic strain of microophthalmic mice (mi/mi), will be used to study the patterns of somatostatin synthesis and expression in the abnormal state, both in vivo and in vitro.

[unreadable] DESCRIPTION (provided by applicant): [unreadable] [unreadable] The studies outlined in this proposal form a cohesive theme focused on ischemic injury and recovery mechanisms in perinatal hypoxia-ischemia. These projects arose as an outgrowth of the last cycle, but with new and exciting directions for the projects. We will now turn our attention to the long-term repair and recovery mechanisms. Our multi-disciplinary approach is maintained in this renewal with the human research project and three laboratory projects that all inform each other and are supported by two cores. The administrative core provides budgetary oversight, training and data management, while the technology core provides a central facility for both the human and laboratory studies for MR imaging and development as well as a repository for MRI development. In Project 1 "White matter injury as predictor of outcome in neonatal brain injury", we plan to focus on the injury to white matter and to correlate the damage in major white matter pathways with neurodevelopmental outcome. We will utilize short echo 3D MR spectroscopic imaging (SEMRSI) to detect changes in metabolites relevant to the injury process, and we also propose to apply high angular resolution diffusion tensor imaging (HARDI) to detect small changes in water diffusion parameters (diffusivity and anisotropy). In Project 2 "Role of Vascular Endothelial Growth Factor (VEGF) in recovery after ischemic neonatal brain injury", we will test whether VEGF plays a critical role in long-term recovery after neonatal stroke by promoting angiogenesis and neurogenesis. In Project 3 "Effects of polyphenols on neonatal Hypoxia-ischemia (H-l) brain injury", we hypothesize that exposure to polyphenols from pomegranate juice as well as the specific polyphenol, resveratrol, will protect the neonatal brain against the acute effects of H-l. In addition, we hypothesize that delayed injury that occurs in the setting of neonatal H-l including axonal injury and regeneration will also be responsive to polyphenols. In Project 4 "Tissue repair in two models of acute neonatal brain injury", we will focus on the repair process in two distinct models of neonatal brain injury - neonatal bacterial meningitis and transient middle cerebral artery occlusion. The two models share common features during the acute phase, such as tissue inflammation and apoptotic neuronal cell death; very little is known about the repair process in these models. These projects are all relevant to human disease as they will bring us closer to therapies for neonatal brain ischemia. [unreadable] [unreadable] RELEVANCE TO PUBLIC HEALTH: Achieving these goals will ultimately result in improved care and outcome in encephalopathic neonates, and potential therapies aimed at the repair process after neonatal ischemic events. [unreadable] [unreadable] [unreadable]

Specific aims are to describe and define neonatal cocaine intoxication and withdrawal using quantitative cocaine/metabolite concentrations obtained from cord blood and correlating these data with examination of the newborn infants in the first few days of life. Additional aims are to investigate the long term effects of cocaine on the developing visual and nervous system through cohort analysis of 1 month old infants with and without gestational drug exposure.

cocaine; embryo /fetus toxicology; drug withdrawal; neurologic manifestations; overdose; newborn human (0-6 weeks); skin; nicotine; developmental neurobiology; drug abuse; ethanol; drug metabolism; lead poisoning; mother /infant health care; pregnancy; caffeine; embryo /fetus membrane; fetal alcohol syndrome; human subject; urinalysis; blood chemistry;

Neurobiology is in a period of explosive growth and change. The development of new techniques, the application of advances in genetics and immunology, the influx of talented scientists from other areas of biomedical science, and the increasing conviction that the time is ripe for major advances in neurobiology, have led to a period of unprecedented growth. Rapid progress will undoubtedly continue through the next decade and will bring major payoffs. Progress in neurobiological research will lead not only to greater understanding of the brain, but will lay the foundation for new modes of diagnosis, treatment and management of neurological and psychiatric disease. Child Neurology deals with illnesses that are among the most debilitating and expensive, and yet this discipline is among the least touched by the modern revolution in biology. Individuals trained in both clinical and basic neuroscience would provide the most effective bridge between disciplines, and could translate basic neuroscience advances into new approaches for understanding and treating neurological diseases in children. This application proposes to develop such individuals through the creation of a research training program for Child Neurology residents at UCSF. The program would integrate clinical and research activities of selected residents in a coherent plan for career development. An effective training program must provide broad and rigorous exposure to ideas and techniques that transcend the bounds of traditional disciplines. We propose to achieve this goal by developing for each trainee a program that incorporates rich experiences in both basic and clinical neuroscience. Clinical training activities will be carried out in the Division of Child Neurology where trainees will become experienced, thoughtful clinicians able to bring critical thinking to bear on clinical problems. Basic science training will consist of both didactic and research activities to be carried out over a minimum of two years. Research training will be carried out in the laboratory of mentors who are established neuroscientists, representing a variety of disciplines, who are committed to trainee development. Formal and informal didactic activities will be carried out under the umbrella of the UCSF Neuroscience Program. The PI and his Advisory Committee will be responsible for candidate recruitment and trainee selection, monitoring of trainee progress, and support of the trainee's transition to an independent research career. A successful training program will result in the production of a number of highly talented Child Neurologist- neuroscientists who pursue productive investigative careers that impact significantly on neurological disease in children.

DESCRIPTION (adapted from applicant's abstract): Neonatal bacterial meningitis leads to severe brain injury in many affected children. The goal of this research is to elucidate the mechanisms of brain injury. Several lines of evidence indicate that meningitis may involve the generation of reactie oxygen intermediates (ROI) and lead to oxidative injury to the brain. A critical role for ROI in meningitis is supported by findings that a scavenger of ROI (alpha-phenyl-tert-butyl-nitrone, PBN) dramatically protected infant rats with experimental neonatal meningitis from developing neuronal injury and was protective in primary co-cultures of neurons and glia stimulated with bacterial products and excitatory amino acids. In Specific Aim 1, the principal investigator will analyze the generation of ROI in primary cultures of neurons, astrocyotes, or microglia in vitro, after stimulation by fragmented cell walls of group B streptococci, cytokines IL-1 and TNF-alpha, ischemia, or excitatory amino acids. In Specific Aim 2, he will confirm the findings of Specific Aim 1 in his infant rat model by identifying production of ROI in the CNS and the effects of ROI scavengers in limiting ROI production, cytotoxicity, and neuronal injury. In Specific Aim 3, he will attempt to dissect the mechanisms by which ROI produce neuroonal injury, again in the infant rat model, by quantitating the effects of ROI scavengers on meningeal inflammation, cerebral ischemia, nitrotyrosine formation, and glial cell activation.

DESCRIPTION (provided by applicant): Recent data from our laboratory suggest that the developing brain responds differently to hypoxia-ischemia (H-I) than the mature brain, therefore requiring different strategies for neuroprotection. Our preliminary data suggest that the developing brain exposed to H-I is more susceptible than the adult brain to injury caused by free radicals, including nitric oxide and hydrogen peroxide. It is our hypothesis that the neonatal brain is more vulnerable than the mature brain because of a greater susceptibility to oxidative stress. This oxidative stress is due to hydrogen peroxide accumulation and sensitivity. To explore this hypothesis we will show that the immature brain accumulates more hydrogen peroxide than the mature brain after H-I, and that the maturity of the nervous system determines the differential responsivity. We will measure hydrogen peroxide accumulation in vivo and in vitro in response to H-I and correlate this with the location and type of cell death. Since the susceptibility to oxidative stress may be due to inadequate scavenging systems in the immature brain, we will measure glutathione peroxidase activity and localization and assess the effect of overexpression of this enzyme on neurological outcome. We will test whether differences in the regulation of glutathione levels and extent of oxidation between immature and mature brain account for differences in susceptibility to oxidative stress. We hypothesize that one mechanism for increased damage with hydrogen peroxide accumulation may be through the formation of hydroxyl ions as a result of the Fenton reaction. This mechanism is particularly important in the immature brain because of the increased amount of free iron. We will measure the localization and concentration of redox available iron and measure hydroxyl formation and outcome after iron chelation. The significance of the findings lies in the clinical application. Prenatal and perinatal hypoxia-ischemia are major causes of severe handicaps in neurologically impaired children. If blockade of free radicals can diminish perinatal H-I injury, development of specific inhibitors may lead to safe, easily administered therapies that could significantly reduce the burden for families and society in caring for these unfortunate children. The ability to identify therapies specific to the immature brain is of utmost importance and relevance to proper therapy.

DESCRIPTION (provided by applicant): It is the goal of this competing renewal application for the Neurological Sciences Academic Development Award that we continue to train, with the highest rigor, candidates who will succeed in academic child neurology. At UCSF, we have created a program that fosters clinical excellence and rigorous postgraduate scientific training. The NSADA has provided the foundation for our division to grow academically and has allowed us to merge clinical training with scientific discovery. The overall theme of our application is "Mechanisms of neurological diseases presenting in infancy and childhood: phenotype-genotype correlations". We will offer rigorous basic and translational scientific training to young investigators skilled at recognizing the phenotypes of complex childhood neurological diseases. This clinical training will allow them to ask important questions about the complex genetics of disease, and should bring understanding to a variety of diseases where the gene is known but the mechanism of disease is not. Our first candidate will study the phenotype-genotype correlations in Tuberous Sclerosis Complex focusing on a subset of patients who present with infantile spasms. She will use the facilities of the Pediatric Clinical Research Center to form a clinical database for these patients, the resources of the Tuberous Sclerosis Alliance for patient enrollment, and the established expertise of leaders in neuroimaging and cell biology of the malformed proteins, tuberin and hamartin. Additional candidates have been identified who will, upon completion of their clinical training, investigate such disorders as channelopathies and autism. We have included in the grant, as well as the appendices, the rich resources that are provided to these applicants. With well established, nationally recognized and supported training programs to support the didactic phase, coupled with a team of research scientists committed to the career development of child neurologists, we will succeed in bringing forth the next generation of leaders in academic child neurology.

Advisory Committees; Budgets; Clinical Research; Clinical Study; Grant; Investigators; Manuscripts; Preparation; Programs (PT); Programs [Publication Type]; R01 Mechanism; R01 Program; RPG; Research Grants; Research Personnel; Research Project Grants; Research Projects; Research Projects, R-Series; Research Resources; Research Training; Researchers; Resources; SCHED; Schedule; Services; Task Forces; computerized data processing; data management; data processing; programs; signal processing

0-6 weeks old; Address; Apoplexy; Astrocytes; Astrocytus; Astroglia; Blood - brain barrier anatomy; Blood Vessels; Blood capillaries; Blood-Brain Barrier; Brain; Capillaries; Capillary; Capillary, Unspecified; Cell Communication and Signaling; Cell Signaling; Cerebral Ischemia; Cerebral Palsy; Cerebral Stroke; Cerebrovascular Apoplexy; Cerebrovascular Stroke; Cerebrovascular accident; Cerebrum; Chelating Agents; Chelators; Cognitive; Cognitive Disturbance; Cognitive Impairment; Cognitive decline; Cognitive function abnormal; Common Rat Strains; Complexons; Coupling; Data; Disturbance in cognition; ECSF; Encephalon; Encephalons; Endothelial Cells; Epilepsy; Epileptic Seizures; Epileptics; Epoetin; Erythropoietin; Extravasation; Fe element; Fetal Liver Kinase-1; Flk-1 Protein; Flk-1 Receptor Tyrosine Kinase; GFAC; Genes; Growth Agents; Growth Factor; Growth Factors, Proteins; Growth Substances; HIF 1; HIF-1 protein; HIF1; HIF1 protein; Hemato-Encephalic Barrier; Hypoxia; Hypoxia Inducible Factor; Hypoxic; Impaired cognition; In Vitro; Individual; Infant, Newborn; Injury; Intracellular Communication and Signaling; Iron; Ischemia; KDR Tyrosine Kinase; Kinase Insert Domain Receptor; Lead; Leakage; Life; Live Birth; Maintenance; Maintenances; Mammals, Rats; Measures; Mediating; Methods and Techniques; Methods, Other; Middle Cerebral Artery Occlusion; Modeling; Motor; NRVS-SYS; Neonatal; Neonatal Brain Injury; Nerve Cells; Nerve Unit; Nervous System; Nervous System, Brain; Nervous system structure; Neural Cell; Neural Growth; Neural Stem Cell; Neurites; Neurocyte; Neurologic Body System; Neurologic Organ System; Neuronal Growth; Neurons; Newborn Infant; Newborns; Occlusion, Middle Cerebral Artery; Outcome; Oxygen Deficiency; Pathway interactions; Pattern; Pb element; Perfusion; Permeability; Personal Satisfaction; Play; Public Health; Rat; Rattus; Receptor Protein; Recovery; Reporting; Rho-associated kinase; Rho-kinase; Role; Seizure Disorder; Signal Transduction; Signal Transduction Systems; Signaling; Societies; Spillage; Stroke; Techniques; Testing; Time; Up-Regulation; Up-Regulation (Physiology); Upregulation; VEGF Receptor Flk-1; VEGF Receptor KDR; VEGFR-2; VEGFR2; VEGFs; Vascular Accident, Brain; Vascular Endothelial Cell; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factors; Vegf; Week; angiogenesis; biological signal transduction; brain attack; burden of disease; burden of illness; capillary; cerebral vascular accident; cognitive dysfunction; cognitive loss; cognitively impaired; day; disability; disease burden; epilepsia; epileptiform; epileptogenic; erythrocyte colony stimulating factor; functional outcomes; heavy metal Pb; heavy metal lead; hematopoietin; hypoxia inducible factor 1; improved; in vitro Model; in vivo; long term memory; neonate; nerve stem cell; neural progenitor cells; neurogenesis; neuronal; neuronal progenitor; neuronal progenitor cells; neuroprotection; newborn human (0-6 weeks); pathway; postnatal; public health medicine (field); receptor; social role; stroke; vascular; well-being; years of life lost to disability; years of life lost to disease

DESCRIPTION (provided by applicant): Project Summary: It is the goal of this competing renewal application for the Neurological Sciences Academic Development Award that we continue to train, with the highest rigor, candidates who will succeed in academic child neurology. At UCSF, we have created a program that fosters clinical excellence and rigorous postgraduate scientific training. The NSADA has provided the foundation for our division to grow academically and has allowed us to merge clinical training with scientific discovery. The overall theme of our application is "Translating discoveries in neurological diseases of infancy and childhood". We will offer rigorous basic and translational scientific training to young investigators skilled at recognizing the phenotypes of complex childhood neurological diseases. Our specific aims are to continue to offer a structured program for training academic child neurologists for a career in translational research;to foster career development;and to expose candidates to the intellectual environment of UCSF. Our first candidate will perform an evaluation of the phosphoinositide 3'-kinase signaling pathway in pediatric malignant gliomas by setting up in vitro and in vivo xenograft models from pediatric brain tumors. She will use the facilities of the Brain Tumor Research Center and the mentorship of Dr. Daphne Haas-Kogan and Dr. William Weiss, experts in the field. Additional candidates have been identified who will, upon completion of their clinical training, will submit protocols for review by the internal and external advisory committees. We have included in the grant the rich resources that are provided to these applicants. With well established, nationally recognized and supported training programs to support the didactic phase, coupled with a team of research scientists committed to the career development of child neurologists, we will succeed in bringing forth the next generation of leaders in academic child neurology. Public Health Relevance: It is necessary to adequately train the next generation of leaders in child neurology so that the future care of children with neurological disease can be addressed and improved.

DESCRIPTION (provided by applicant): This proposal is the first competing renewal of a grant initially funded in 2003 in response to the RFA HD- 02-019 for NICHD Institutional Training for Pediatricians. Our application requests ongoing resources to train 4 pediatricians with an M.D. or M.D./PhD. degree each year in pediatric research in the Department of Pediatrics at the University of California San Francisco (UCSF). In the first 4 years of this award we have supported the training of 12 pediatric subspecialty fellows (4 men and 8 women). All 5 trainees who have finished their fellowships have academic appointments (4 at UCSF) and two-thirds of the fellows with at least three years of post-doctoral training have submitted or hold a career development award (K award). The Department of Pediatrics at UCSF is strongly committed to producing young pediatric researchers who are passionate about translating scientific data to enhance the health of children. The goal therefore of this T32 application is to provide rigorous scientific training and extensive career mentoring to pediatric subspecialty fellows who are committed to a sustained career in research is highly relevant to pediatric disease. We anticipate that most scholars will emphasize either Tl (Laboratory to Human Subjects) or T2 (Evidence to Practice) research and outline appropriate training components specific to each pathway. However, we also recognize that some trainees will pursue projects that will integrate elements of Tl and T2 investigation or that research and career goals may shift during the period of T32 support. For this reason, the program is designed to work proactively with each T32 scholar to design and implement an optimal training program. The training program for Tl and T2 fellows consists of the following four key elements: (1) a mentored research experience;(2) scientific course work;(3) interdisciplinary experiences and career development workshops;and, (4) ongoing review and evaluation. A well-established mentoring program is in place. The objective goals of the T32 program are to have 90% of the graduates enter academic medicine at the completion of training (actual performance to date 100%), 80% remain in academic medicine for greater than 5 years, two-thirds of the graduates apply for a K award within 12 months of completing training (actual performance to date 63%), and 50% of graduates to have an independent, federally-funded grant within 5 years of graduation (not enough time has passed to evaluate performance on this metric).

DESCRIPTION (provided by applicant): This revised proposal is submitted in response to RFA-HD-13-011, Child Health Research Career Development Award (CHRCDA) Program to provide K12 awards through the CHRCDA mechanism to young pediatric investigators. The application requests resources to support four pediatricians each year who hold MD or MD/PhD degrees and have completed scholarship training in a clinical subspecialty. The rationale for the program is based on the well-documented and urgent need to support mentored career development for pediatricians to enable them to become fully independent and productive basic science researchers, and the fact that UCSF has the vision, experience and infrastructure to train the next generation of leaders in pediatric science. Our aims are to (1) offer a structured program for training academic pediatricians, (2) foster career development and promote retention of junior faculty, (3) expose promising early career pediatricians to the intellectual richness of UCSF research and (4) promote diversity in academic pediatrics. The scholars trained by this program will bring state-of-the-art approaches to bear on diagnosis, treatment and prevention of health problems in children as well as childhood onset of adult illness. The design of this program involves harnessing the expertise of world-class basic laboratory scientists who will serve as mentors for interdisciplinary training. The basic science training program is focused around seven scientific cores: cancer, cardiopulmonary medicine, developmental biology, genetics, immunology, neurobiology, and stem cell biology. Each core has a Director, designated faculty, and a specific didactic curriculum. The scholars, in conjunction with their mentor and Core Director, will also participate in a program of additional discipline-specific course work dependent on both the prior experience and training of the applicant and the scientific theme of the trainee's research, which may often overlap amongst different cores. We believe that the Department of Pediatrics at UCSF has the vision, experience, and infrastructure to train the next generation of leaders in academic pediatrics. In this application, we provide evidence that the Department together with the broader UCSF research community comprise an exceptional environment for preparing young pediatricians who will receive support through the CHRCDA mechanism for successful careers as basic science researchers. For example, there are over 1,000 research laboratories and over 2,200 active research projects at UCSF, and the faculty currently includes 4 Nobel laureates, 41 National Academy of Sciences members, 61 American Academy of Arts and Sciences members, 72 Institute of Medicine members, and 16 Investigators of the Howard Hughes Medical Institute. This program is an investment in the future of children's health, as the diverse group of researchers we will train will harness advanced research strategies to address urgent problems (e.g., asthma, cancer, infectious diseases, host defense defects, diabetes, obesity, and developmental malformations of the heart, brain, and other organs), that will result in new treatments to improve child health outcomes.

The core component functions as an administrative and coordinating center for all the components of the program project. The administrative functions include program oversight, including budget, management and planning, data processing for the clinical research projects and manuscript preparation. It will serve to facilitate interactions among the investigators in the center as well as coordinate activities with the internal and external advisory committees. The aim ofthe administrative core is to function as a centralized facility where administrative resources can be utilized to provide support and integration of services for all investigators. The specific aims of the administrative core will be to function as a resource for grants administration, data management and analysis, research training opportunities and a seminar series. The administrative committee will make decisions regarding the use of Core Services. This committee is chaired by the program director and is composed ofthe principal investigators of each project and include Drs Barkovich, Xu, and McQuillen . This group will meet at regular intervals regarding administrative and scientific matters such as facilities management, personnel matters data sharing, and will share responsibility for scientific seminars. In addition, the project leaders and investigators meet weekly on Wednesday to discuss issues of scanning, nursing needs, screening and other issues pertinent to the enrollment of subjects. For any adjudication. Dr. Ferriero will decide when there is incomplete agreement on matters that relate to the overall program. Cost-effectiveness is achieved through budgetary oversight, a single administrator who directly communicates with the program director and the project leaders. Quality control is provided as a service ofthe Department of Pediatrics Core Financial Unit through the internal auditing of budgets and processes. Additional quality control is realized through the use of our internal and external advisory committees.

DESCRIPTION (provided by applicant): This grant proposes to study both structural and functional correlates of brain developmental maturation and network organization using advanced imaging techniques in our human populations and similar correlates in newborn rodents with a focus on defining basic mechanisms of repair. The grant will encompass three projects and two cores: Project I) repair after ischemic injury in the term newborn - using both structural and metabolic serial imaging to evaluate how the injuries have altered the brain, its response to injury in the perinatal period, and how structural connectivity relates to neurodevelopmental outcome. This cohort will include babies treated with hypothermia. Project II) repair in high risk newborns with congenital heart disease- there is delayed brain development in this high risk population. Using structural and metabolic imaging, we will determine whether this disorder of development is associated with abnormalities in brain connectivity and functional outcome. Project III) repair after early preterm birth - our data show many preterm infants to have abnormalities of cerebellar maturation, abnormalities of white matter development or progressive white matter disease with infection. Using structural and metabolic imaging, we will determine whether altered patterns of connectivity are consistent or variable, whether recovery from these insults is possible and whether connectivity is permanently or transiently disrupted by these early ex-utero life events. There is an administrative Core for data management including biostatistical support, budgetary oversight, training and seminars. The Imaging and Neurobehavior Core (Services Core) will support the three human imaging projects by ensuring standardization of methods and tracking of imaging acquisition at the various testing time points. In addition, this Core will provide Neurobehavioral testing at all of the follow-up visits, and will be responsible for tracking patients for all projets.

Project Summary/Abstract Section Neonatal brain injury is an important cause of death and disability, with pathways of oxidant stress, inflammation, and excitotoxicity leading to damage that progress over a long period of time. Therapies have classically targeted individual pathways during early phases of injury, but targeting pathways later in the injury response may be additionally effective. Therapeutic hypothermia (TH), while being standard of care for hypoxic-ischemic encephalopathy (HIE), provides protection only in 60% of babies. The overarching hypothesis is that the metabolic state of the brain immediately after TH differs markedly between hypothermia responders and non-responders. We will identify the metabolic state after TH using proton and hyperpolarized carbon 13 spectroscopy and then study cellular pathways to identify more precise and individualized treatment approaches. We will use postnatal day 9 mice and follow them through injury evolution. The studies outlined in this proposal reflect an innovative and systematic approach to the study of HI brain injury in the newborn because they combine advanced metabolic imaging techniques (proton and carbon spectroscopy) and cell-signaling studies, focusing on HIF signaling, that will both inform and be informed by human clinical studies. Utilizing genetic cellular approaches in which components of the injury response are specifically deleted/disrupted in the specific cell compartments will allow us to evaluate the benefits of the neurovascular niche in vivo. We will broadly interrogate HIF-dependent signaling pathways following injury using ChIP-Seq. Together, these genetic tools will allow us to explore the molecular regulation of HI both in vitro and in vivo to better identify more appropriate molecular targets for therapy for the individual needing them the most. By investigating responses to HI at a cellular level using traditional biochemical assays and global level in the brain using MR spectroscopy, we aim to make a link between specific cellular changes and metabolic changes that can be detected non-invasively. This approach would eventually allow the findings to be translated into the clinic and potentially change the management of patients. We will use in vitro techniques such as CRISPr/Cas9 and Chip-seq to dissect important signaling pathways like hypoxia inducible factor (HIF). We will also use an invitro approach when we identify appropriate targets and design therapies to counteract deficient repair. Thus, defining the cerebral metabolic signature of non- responders will identify subsequent novel pathways to target, and will lead to improved outcomes that could never be achieved by only targeting pathways through hypothermia alone. Understanding how the cascade of injury responses occur and the key modulators during each phase will lead to more rationale therapies.

PROJECT SUMMARY/ABSTRACT The Hershey Conference on Developmental Brain Injury was started in 1997 and has grown to a popular, well attended, yet still small, international conference of physician-scientists and basic scientists, with a high number of trainees. The 2018 11th Hershey Conference will return to the USA and will be organized by Drs. Donna Ferriero and Susan Vannucci to be held at Asilomar CA. We are applying for support from NIH primarily to fund attendance for junior investigators and trainees. The overall goal of the proposed meeting is to bring together an international group of clinical and basic scientists involved in research pertaining to brain injury and regeneration in developing animals and humans. The approach remains the same as previously and has four objectives: 1) identifying clinical paradigms that inform basic science concepts and translating those new basic concepts into more refined clinical application; 2) exploring areas of gaps in our current knowledge and including experts in tangential fields to provide new perspectives on these gaps 3) promoting novel collaborations 4) recruiting and mentoring of junior colleagues, especially including under-represented minorities Therefore, the objective of this R13 is to provide the necessary continued support of trainees and junior faculty. Extensive participation of trainees, both from US and abroad, has been a hallmark of the Hershey Conference. Trainees, both MD and PhD, constitute 30-50% of all participants. This meeting has been an excellent opportunity for informal interaction between trainees and the leaders in the field as well as a mechanism for post-doctoral and visiting scientist recruitment. Thus, a specific aim of this R13 is to promote trainee and junior faculty participation and to identify trainees/junior faculty from racial/ethnic minorities and underrepresented institutions to attend this conference.

ABSTRACT The Hershey Conference on Developmental Brain Injury was started in 1997 and has grown to a popular, well attended, yet still small, international conference of physician-scientists and basic scientists, with a high number of trainees. The 2020 12th Hershey Conference will return to Europe and will be organized by Drs. Henrik Hagberg, Carina Mallard from Sweden and Drs. Ferriero and Vannucci from the US. We are applying for support from NIH primarily to fund attendance for junior investigators and trainees. The overall goal of the proposed meeting is to bring together an international group of clinical and basic scientists involved in research pertaining to brain injury and regeneration in developing animals and humans. The approach remains the same as previously and has four objectives: 1) The mentoring of trainees and junior colleagues with the specific goal of keeping them engaged in academics 2) The strengthening of existing collaborations and the promotion of novel collaborations especially with investigators from outside the field 3) Exploration of specific areas where there are gaps in our current knowledge 4) Identification of advanced techniques that will rapidly move the field forward Therefore, the objective of this R13 is to provide the necessary continued support of trainees and junior faculty. Extensive participation of trainees, both from US and abroad, has been a hallmark of the Hershey Conference. Trainees, both MD and PhD, constitute 30-50% of all participants. This meeting has been an excellent opportunity for informal interaction between trainees and the leaders in the field as well as a mechanism for post-doctoral and visiting scientist recruitment. Thus, a specific aim of this R13 is to promote trainee and junior faculty participation and to identify trainees/junior faculty from racial/ethnic minorities and underrepresented institutions to attend this conference.