Jacqueline C. Bresnahan - US grants

The Neurotrauma Society Meeting is held annual as a satellite symposium of the Society for Neuroscience Meeting, this year in LA. The theme for this fall s meeting is Recovery from Neurotrauma, from molecules to man . The meeting lasts for 2 full days, just before the NS meeting, and is balanced between lectures and posters (four sessions, grouped by topic, with assigned times for author presentation). There are 5 regular lecture sessions (1 in the morning and 2 in the afternoon) where internationally recognized (and hence in general more senior investigators) give 20 minutes talks with 10 minute of discussion. Poster sessions occur as one hour breaks in each morning and afternoon schedule. To provide a more open format, there are also 2 Open Communication Sessions where 12 speakers will be chosen from submitted abstracts by the program committee and more junior investigators will have a chance to present their material orally (10 minute talks with 5 minute discussions). The top 18 student posters will be judged for a special student poster competition (with two cash prizes contributed by industry). Abstracts will be made available on the Web, so that participants can schedule the posters they wish to visit in advance, and the proceeds will be published in the Journal of Neurotrauma, which is the official journal of both the national and international chapters of the Neurotrauma Society. A recent letter from the PI has confirmed all of the invited speakers and session chairs. This meeting has been held annually since 1982 and has grown over the years so that last year there were 15-20 invited speakers, 200 posters, and 350-400 attendees. Only 1/3 of the society members are also members of the SN, and attendees include a large number of clinicians, for whom this is a chance to learn what new basic research findings have been made in this field. A major aim of the society is to encourage students and postdocs in this area. This is done via travel grants, awards for the best submitted abstracts, and the opportunity to present their work as a short lecture for the best abstracts. Another aim is to encourage participation of qualified women and minorities in neurotrauma research. To this end, there is newly chartered Women in Neurotrauma Research (WINTR) which hosts a panel discussion during lunch of the first day (past years: opportunities for funding and a mock study section, this year a discussion of how to integrate research with family obligations for women and for dual career couples in general).

The final functional outcome of spinal cord injury (SCI) is the result of an interplay between secondary degenerative events and endogenous mechanisms of repair. We now know that the secondary injury process includes the induction of programmed cell death in both glia and neurons. We also know that there are substantial reparative events in the cord after injury that involve growth and trophic factors. Exciting new information on the stimulation of proliferating progenitor cells after injury to the CNS suggest that more repair may be possible than previously imagined. We have developed models of contusion spinal cord injury in rats that mimic some of the clinical features of SCI in man. Recovery occurs over time in the face of continuing apoptotic cell death. At the same time, ependymal zone cells as well as cells in the white proliferate and appear to contribute to repair at the lesion margins. In addition, new axonal growth can be seen entering the lesion area. Using these observations as a background, we will examine the effects of methylprednisolone (MP) and basic fibroblast growth factor (bFGF) on these cellular events after SCI. MP is the current Agold standard@ of clinical treatment of acute SCI, but its mechanisms of action are not fully understood. BFGF is a promising treatment for brain and spinal trauma that is likely to retard apoptosis and drive cellular proliferation of CNS progenitor cells. We will study recovery of function and the effects of MP and bFGF after both contusion and dorsal hemisection SCI using established behavior methods. We will examine the effect of these agents on apoptotic cell death of neurons and glia, and on cell proliferation and phenotypic expression. We will examine the effects of combination treatments on behavior and the regeneration of the corticospinal tract, which will be used a barometer of axonal growth in these two lesion types. The results are expected to provide further information on the biology of SCI as well as an assessment of potential therapies that could reach the clinic in a short time.

Inflammation and excitoxicity appear to be pivotal in CNS trauma, but effective strategies for targeting these aspects of secondary injury have been elusive. Proinflammatory cytokines appear to have complex and sometimes contradictory roles in CNS injury and repair. Similarly, glutamate receptors are responsible for excitotoxic death of neurons and glia in injury, but are also essential for normal CNS function, and have been implicated in recovery after injury. The pro-inflammatory cytokine tumor necrosis factor-alpha (TNFa) has recently been shown to have a unique and critical role in the modulation of normal neuronal glutamate synaptic transmission (Beattie et ai, 2002;Stellwagen and Malenka, 2006;Aizenman and Pratt, 2008), and also to exacerbate excitotoxic cell death (Hermann et ai, 2001;Beattie, 2004). We have identified TN Fa-mediated trafficking of GluR2-lacking, Ca++-permeable AMPA receptors (CP-AMPARs) as a novel and perhaps 'nodal'link between injury-induced inflammation and excitotoxicity. TN Fa increases excitotoxic cell death in vitro, and enhances neuronal death after spinal cord injury (SCI). Further, reducing GluR2- lacking AMPAR-insertion into neuronal membranes by blocking TNFa after SCI, results in reduced neuronal death, reduced white matter damage, and better outcomes in a cervical injury model of SCI.

DESCRIPTION (provided by applicant): The innate immune response is clearly an integral part of the secondary injury cascade in CNS trauma, but the role of glial and monocyte derived pro-inflammatory cytokines is complicated by multiple downstream effects mediated by multiple concentration-dependent receptors, and a rapidly changing and evolving microenvironment. We have new findings that strongly support the hypothesis that TNF and AMPAR changes are critical in both secondary injury and recovery after CNS injury. Sequestering TNF using soluble TNF receptor protein (sTNFR1) reduces damage after cervical SCI, and, in a highly dose-dependent manner, improves neurological outcomes. We propose to extend our studies of TNF and AMPAR trafficking as a therapeutic target for SCI using a multivariate approach to test preclinical efficacy. Etanercept (ETAN) is a TNF-sequestering protein biologic used clinically in rheumatoid arthritis;topiramate (TPM) is a neuroprotective, anti-epileptic drug that has AMPAR antagonism. We will use these drugs to target AMPAR-trafficking and AMPAR activity, separately and in combination. We will systematically evaluate the biological responses to these drugs using early post-injury biomarkers that have predicted neurological outcomes in our prior work. We will use the biomarker data to plan preclinical dose and timing regimens to evaluate effectiveness in both cervical and thoracic rat SCI, and evaluate their effects on autonomic, sensory and motor outcomes. These efforts are aimed at moving towards clinical application of anti-TNF therapies for SCI, and may be applicable to other CNS degenerative disorders as well. We propose three specific aims: Aim 1: We will evaluate the effects of ETAN and TPM on the time course and extent of biomarkers of AMPAR surface expression, cell death and the production of pro-inflammatory cytokines after cervical SCI. We predict that these treatments will reduce the feed-forward cascade of cell death. Aim 2. Guided by these data, we will optimize dose and timing of single and combination therapies to maximize six week recovery after unilateral cervical SCI using a variety of forelimb functional tests (grooming, paw placement, Catwalk and IBB). Aim 3: We will test the effects of optimized drug regimens on recovery from thoracic contusion lesions using a battery of tests that includes autonomic, sensory, and motor outcomes. This will establish whether efficacy extends to multiple models of SCI. TNF and AMPARs are also involved in the production of chronic hypersensitivity after nerve injury (Choi et al, 2010), and we will test whether this occurs after SCI as well. TPM is already used to treat chronic SCI pain. We predict that acute treatments with ETAN and TPM that result in better motor outcomes will also result in reductions in long term allodynia and in tonic, aversive central pain (King et al, 2009), the latter measured by place preference tests. PUBLIC HEALTH RELEVANCE: This project is aimed at the preclinical development of therapies for spinal cord injury (SCI). The drugs we will use target a unique aspect of the 'secondary injury'process: namely the role of tumor necrosis factor in altering the effects of a neurotransmitter, glutamate, on neurons and glial cells. Some of these drugs are already approved for other indications (e.g. etanercept and topiramate), so progress to clinical applications could be rapid, leading to better recovery after SCI and savings of health care costs.

This is a collaboration between experts at UCSD, UCLA, UCSF, UC Davis, UCI and the Ecole Polytechnique Federale de Lausanne (EPFL) to examine plasticity and regeneration in the non-human primate spinal cord. Our goal is to enhance knowledge and translational relevance of research on spinal cord injury (SCI). This project will map the motor cortex ?connectome? to better understand the primate motor network in both the intact state and after recovery from SCI, and will focus on promoting therapeutic growth of the most important motor control system in primates, the corticospinal projection. Aim 1: The Intact Primate Corticospinal Connectome Aim 1 will map the intact connectome for motor control of the hand using new generation, highly specific and highly sensitive viral tools. These findings will be correlated with motor cortex recordings and forelimb EMG activity in awake, behaving subjects. These tools will allow an unprecedented understanding of hand motor control in the primate, thereby revealing novel mechanisms of motor control and identifying new targets for therapy. Aim 2: The Lesioned Primate Corticospinal Connectome How does injury affect the corticospinal connectome? How does the corticospinal system adapt to injury and alter its set of outputs, and how does this influence functional recovery? We will use the elegant and novel tools of Aim 1 to map the injured, reorganized corticospinal connectome, motor cortex dynamics, and forelimb EMG after C7 hemisection lesions. Aim 3: Spinalized Neural Stem Cell Grafts to Enhance Corticospinal Repair Work in Aim 3 will build on recent progress in neural stem cell technology. We will use ?spinalized? neural stem cells to augment growth of the injured corticospinal tract and form trans- lesion relays that enhance recovery of forelimb function after SCI. Further, we will compare the connectome of the corticospinal system after therapeutic stimulation to its intact and lesioned state. This work will reveal how new corticospinal growth after injury impacts the projections and connections of this vitally important motor system in humans. 1

DESCRIPTION (provided by applicant): Neurotrauma is a major public health problem for which there are currently no effective therapies. Neurotrauma is the largest cause of death and disability, for persons under age 45 in the world, and the societal costs of the resulting disabilities exceeds $50 billion per year. The National Neurotrauma Symposium, now in its 32nd year, has been the primary forum for exchanging information on research in the fields of both traumatic brain injury (TBI) and spinal cord injury (SCI) for many years. The meeting focuses on integrating basic, translational, and clinical scientific information on neurotrauma. The 2014 meeting, co-hosted by the National Neurotrauma Society (NNS), and the AANS/CNS Joint Section on Neurotrauma and Critical Care (AANS/CNS JSNCC), will be held in downtown San Francisco, CA at the Marriott Marquis Hotel on June 29-July 3. This application requests support for 20 pre-and post-doctoral trainees to attend the meeting and partial support for travel for the 57 speakers. An exciting collation of state of the art clinical, translational and basic science sessions on the consequences of damage to the nervous system, and treatment strategies for repair after injury is planned. Highlights of the meeting include sessions on chroni traumatic encephalopathy and sports concussions, pediatric TBI, cell transplantation for SCI (both clinical trials and recent exciting lab results showing enhanced recovery), synaptogenesis after neurotrauma, rehabilitation strategies in both the lab and clinic for SCI and TBI, a roundtable discussion of model selection, pain after neurotrauma, and novel approaches for future basic and clinical research. There will be mentoring and networking sessions organized by the Women in Neurotrauma Research (WiNTR) for students and young investigators, and a special symposium day devoted to Neurotrauma Allied Health Professionals. A public lecture with Bob Woodruff will be held on the first day of the meeting, and an airing of The Crash Reel, a documentary about Kevin Pearce an Olympic snow-boarding contender who suffered a head injury, is planned to provide important patient and family perspectives.

? DESCRIPTION (provided by applicant): This proposal represents an interdisciplinary team that has tremendous potential to impact trauma patient care. San Francisco General Hospital (SFGH) is one of four main campuses comprising UCSF, and it is the only Level 1 Trauma Center that serves the city of San Francisco. Research at SFGH is varied, but includes a large investment in trauma-related research both clinically and experimentally. However, until now, there has not been a concerted effort among basic scientists to leverage individual expertises and develop integrated animal models that correspond to commonly observed clinical polytrauma conditions. The interdisciplinary team assembled here represents a team of Neurosurgery and Orthopaedic Surgery researchers and clinicians that have collaborated to address the clinical challenges associated with understanding and treating polytrauma. For this proposal we have developed a mouse model of combined brain and bone injury in order to study the mechanisms by which injury in one tissue affects healing in another tissue. We chose to combine a traumatic brain injury (TBI) with a long bone fracture because this is a common combination of injuries observed during high velocity traumas. Furthermore, clinical data shows that brain injury accelerates fracture healing and increases incidence of heterotopic ossification, but the mechanism underlying this altered repair dynamic is unknown. While the description of these phenomena are engrained in the clinical literature, animal models to examine this combination of injuries are not common as they require expertise in divergent fields. The research proposed here will promote valuable interdisciplinary collaborations within our institution. As indicated, we chose to pair brain and bone injuries as an initial project, because there are well-described clinical outcomes for which the underlying molecular mechanisms are not understood. However, SFGH has a multitude of researchers studying a large variety of trauma including brain injuries and spinal cord injuries, cutaneous wounds, gastrointestinal and lung injuries. All of these models are currently being studied as isolated injuries, which is not commonly the clinical condition. To address this we have created the SFGH Polytrauma Research Initiative which is committed to expanding our understanding of molecular basis underlying combined injuries that occur during trauma in order to accelerate improvements in clinical practice to treat polytrauma patients. These goals are directly in line with the mission o NIAMS.

PROJECT SUMMARY/ABSTRACT This is a new multi-PI proposal from our California Spinal Cord Injury (SCI) consortium to continue to translate exciting results from the transplantation of neural stem cells (NSCs) from rodent to primate, and to evaluate efficacy and safety in our non-human primate (NHP) cervical contusion SCI model. Our multi-center consortium has examined recovery of function and its anatomical correlates in a series of studies using a cervical hemisection model. We have discovered spontaneous and extensive plasticity of the corticospinal tract (CST) system that had not been appreciated in previous rodent studies. We have developed the first large NHP model of cervical hemicontusion SCI together with an open-field scoring system and novel in-cage forelimb activity and hand function tests to evaluate functional outcomes. The wealth of new information and directions speak to the value of this shared approach to using the very valuable primate model. This project focuses on translation of our NHP stem cell work. We now report that neural stem cells (NSCs) derived from human spinal cord grafted early to hemisection sites in NHP SCI, extend very large numbers of axons over very long distances, and that these transplants appear to enhance long-term recovery of hand function, and support CST regeneration into the graft. NSCs derived from the approved human embryonic stem cell H9 (H9 hESCNSCs) also support CST regeneration into spinal cord grafts in the NHP after SCI. Further, we have advanced our cell therapy strategy to produce the first H9 hESCNSCs caudalized to move them towards a spinal cord fate, and have shown that transplants of these cells in rodents promote much more vigorous regeneration of CST axons7. Therefore, in this proposal in NHPs, we will transplant caudalized hESCNSCs into a contusion lesion at a more chronic and clinically relevant six week time point. We hypothesize that these grafts will support robust CST regeneration and enhance recovery of forelimb function, and provide a relay for CST axons to influence forelimb circuitry in the C8-T1 cord. We will use anterograde and retrograde tracing, IHC and transfection of graft cells and correlate the connectional data with recovery, and test the long-term survival, safety, and functional effects of these transplants.

PROJECT SUMMARY: Trauma to the spinal cord and brain (neurotrauma) together impact over 2.5 million people per year in the US, with economic costs of $80 billion in healthcare and loss-of-productivity. Yet precise pathophysiological processes impacting recovery remain poorly understood. This lack of knowledge limits the reliability of therapeutic development in animal models and limits translation across species and into humans. Part of the problem is that neurotrauma is intrinsically complex, involving heterogeneous damage to the central nervous system (CNS), the most complex organ system in the body. This results in a multifarious CNS syndrome spanning across heterogeneous data sources and multiple scales of analysis. Multi-scale heterogeneity makes spinal cord injury (SCI) and traumatic brain injury (TBI) difficult to understand using traditional analytical approaches that focus on a single endpoint for testing therapeutic efficacy. Single endpoint-testing provides a narrow window into the complex system of changes that describe the holistic syndromes of SCI and TBI. In this sense, complex neurotrauma is fundamentally a problem that requires big- data analytics to evaluate reproducibility in basic discovery and cross-species translation. For the proposed TOP-VISION cooperative agreement we will: 1) integrate preclinical neurotrauma data on a large-scale; 2) develop novel applications of cutting-edge multidimensional analytics to make sense of complex neurotrauma data; and 3) validate bio-functional patterns in targeted big-data-to-bench experiments in multi-PI single center (UG3 phase), and multicenter (UH3 phase) studies. The goal of the proposed project is to develop an integrated workflow for preclinical discovery, reproducibility testing, and translational discovery both within and across neurotrauma types. Our team is well-positioned to execute this project given that with prior NIH funding we built one of the largest multicenter, multispecies repositories of neurotrauma data to-date, housing detailed multidimensional outcome data on nearly N=5000 preclinical subjects and over 20,000 curated variables. We will leverage these existing data resources and apply recent innovations from data science to render complex multidimensional endpoint data into robust syndromic patterns that can be visualized and explored by researchers and clinicians for discovery, hypothesis-generation and ultimately translational outcome testing.

PROJECT SUMMARY: Trauma to the spinal cord and brain (neurotrauma) together impact over 2.5 million people per year in the US, with economic costs of $80 billion in healthcare and loss-of-productivity. Yet precise pathophysiological processes impacting recovery remain poorly understood. This lack of knowledge limits the reliability of therapeutic development in animal models and limits translation across species and into humans. Part of the problem is that neurotrauma is intrinsically complex, involving heterogeneous damage to the central nervous system (CNS), the most complex organ system in the body. This results in a multifarious CNS syndrome spanning across heterogeneous data sources and multiple scales of analysis. Multi-scale heterogeneity makes spinal cord injury (SCI) and traumatic brain injury (TBI) difficult to understand using traditional analytical approaches that focus on a single endpoint for testing therapeutic efficacy. Single endpoint-testing provides a narrow window into the complex system of changes that describe the holistic syndromes of SCI and TBI. In this sense, complex neurotrauma is fundamentally a problem that requires big- data analytics to evaluate reproducibility in basic discovery and cross-species translation. For the proposed TOP-VISION cooperative agreement we will: 1) integrate preclinical neurotrauma data on a large-scale; 2) develop novel applications of cutting-edge multidimensional analytics to make sense of complex neurotrauma data; and 3) validate bio-functional patterns in targeted big-data-to-bench experiments in multi-PI single center (UG3 phase), and multicenter (UH3 phase) studies. The goal of the proposed project is to develop an integrated workflow for preclinical discovery, reproducibility testing, and translational discovery both within and across neurotrauma types. Our team is well-positioned to execute this project given that with prior NIH funding we built one of the largest multicenter, multispecies repositories of neurotrauma data to-date, housing detailed multidimensional outcome data on nearly N=5000 preclinical subjects and over 20,000 curated variables. We will leverage these existing data resources and apply recent innovations from data science to render complex multidimensional endpoint data into robust syndromic patterns that can be visualized and explored by researchers and clinicians for discovery, hypothesis-generation and ultimately translational outcome testing.