Anthony Kline - US grants

DESCRIPTION (provided by applicant): Traumatic brain injury (TBI) affects 1.5 to 2 million individuals in the United States each year. Approximately 30% of the 300,000 severely injured survivors endure long-term disabilities, leading to costly medical and rehabilitative care. In addition to the physical impairments, TBI survivors also face a significant disturbance in cognitive function. Because memory impairment is a multifaceted phenomenon, alleviating this dysfunction may necessitate either a combination of pharmacotherapies or single pharmacologic agents that modulate various neurotransmitter systems. In this application, the investigators are investigating the latter possibility via a serotonin (5-HT-1A) receptor agonist -- a phamacotherapy paradigm used clinically to treat depression and anxiety. Both the 5-HT and acetylcholine (ACh) neurotransmitter systems are implicated in mediating cognition. 5-HT-1A receptor (5-HT-1AR) agonists are known to interact with the cholinergic system by increasing ACh release. ACh neurotransmission is chronically decreased after TBI, which may, in part, contribute to cognitive deficits. Thus, the interaction between the serotonergic and cholinergic systems via 5-HT-1AR agonists may provide an alternative and potentially beneficial therapeutic strategy after brain injury. Preliminary support for this hypothesis stems from recent work demonstrating that acute 5-HT-1AR agonism attenuates learning and memory deficits produced by controlled conical impact injury. While the mechanisms for the beneficial effect observed in this novel therapeutic approach are not known, the results have prompted a general hypothesis that 5-HT-1AR agonism attenuates TBI-induced cognitive dysfunction by increasing cholinergic neurotransmission and decreasing cholinergic cell death. The following aims are proposed to test this hypothesis: Aim I will evaluate the potential efficacy of a delayed (24 hr after injury) and chronic (20 days) 5-HT-1AR agonist treatment paradigm on cognitive performance after experimental TBI produced by a well-established cortical impact model. Aim 2 will assess biochemical and immunohistochemical markers of cholinergic neurotransmission after chronic 5-HT-1AR agonist treatments. The results from these novel studies will demonstrate for the first time the effects of chronic treatments with a 5-HT-1AR agonist on cognitive performance after TBI as it relates to a rehabilitative setting. This R03 pilot project will serve as proof of concept for the basis of a more comprehensive study of 5-HT-1AR-mediated mechanisms after TBI. The long-term goal is to develop pharmacotherapies that attenuate posttraumatic neuronal cell death and facilitate cognitive performance after human TBI.

DESCRIPTION (provided by applicant): Traumatic brain injury (TBI) affects 1.5 to 2 million individuals in the United States each year. Approximately 100,000 severe-TBI survivors endure long-term memory and/or physical impairments that require rigorous and costly rehabilitative therapy. While there are currently no accepted treatments for human TBI, we have data showing that early (15 min after TBI) administration of serotonin/lA receptor (5-HT1AR) agonists - a pharmacotherapy novel to TBI, but used routinely to treat anxiety and depression in humans - attenuate experimental TBI-induced behavioral deficits. While the benefits of this early treatment are compelling, the potential efficacy of delayed and chronic 5-HT1AR agonist treatments after TBI is unknown. This issue is paramount given the secondary sequelae that are prevalent hours to days after TBI and that perturb the recovery process. Empirical investigation of pharmacological interventions that restore and/or enhance neuromodulation when given hours vs. minutes post-TBI is essential for successful rehabilitation. Also warranted is further investigation of environmental enrichment (EE) on recovery. EE enhances outcome after TBI vs. standard environments, is a relevant experimental analogue of the rehabilitation paradigm, and may provide clinical utility alone or as an adjunct to pharmacotherapy. Thus, the goals of this proposal are to further examine the effects of the 5-HT1AR agonist buspirone and EE alone or in conjunction with each other. It is hypothesized that huspirone and EE will facilitate memory and motor recovery after TBI, but the combination of treatments will be more efficacious than either alone. To test this hypothesis, a logical series of aims are proposed: Aim 1 will investigate the potential efficacy of a delayed and chronic treatment regimen (1-20 days post-injury) with buspirone on functional recovery after TBI produced by a well-established cortical impact injury model that produces deficits resembling those seen clinically. Aim 2 will determine the effects of EE on functional outcome after TBI. Aim 3 will investigate the effects of EE plus buspirone treatment on recovery after TBI. Both treatments augment cholinergic function, which is strongly implicated in memory and decreased after TBI, and thus Aim 4 will explore this avenue as a potential mechanism by quantifying biochemical and immunohistochemical mediators of cholinergic neurotransmission. This project represents the first systematic investigation of delayed and chronic 5-HT1AR agonist treatments, EE, and their combination on memory and motor function after TBI, and will provide a framework for further empirical research of potential mechanisms. Our long-term goal is to develop therapies that facilitate functional recovery after human TBI.

DESCRIPTION (provided by applicant): Exposing rats to complex, stimulating, and social housing (i.e., enriched environment; EE) improves cognitive and motor performance after traumatic brain injury (TBI). The specific mechanisms that underlie this effect are unknown, although there is evidence suggesting that EE enhances cortical plasticity. Furthermore, subcortical cholinergic afferents are critically involved in eliciting specific forms of plasticity in rats, such as estrogen-mediated plasticity in the hippocampus, denervation-mediated plasticity in the frontal cortex, and experience-related plasticity in both visual and auditory cortices. The proposed studies will test the hypothesis that cholinergic neurons are important mediators of EE-induced change after TBI and that there are gender differences in this mediation. As such, cholinergic enhancement may be an effective means of augmenting the beneficial effects of `ancillary' treatments, such as behavioral enrichment (i.e., rehabilitation), in patients with TBI. Five specific aims are proposed to test the overall hypothesis. Aim 1 will determine whether disrupting basal forebrain cholinergic neurons (BFCNs) blunts the ability of EE to improve behavioral performance after TBI produced with the well-established controlled cortical impact injury model in conjunction with saporin (SAP)-lesions. Aim 2 will determine to what extent EE-induced synaptic plasticity in these same animals (i.e., Aim 1) is altered and if plasticity correlates with performance. Because hormones are known to modulate cholinergic function and males and females respond differently to EE after TBI, Aim 3 will assess cortical plasticity and functional improvement with EE in gonadally intact vs. ovariectomized (OVX) female rats after TBI+/-SAP lesions and compare the results to males from Aims 1-2. Aim 4 will assess the effect of a delayed and chronic treatment regimen (1-20 days post-injury) with the FDA-approved acetylcholinesterase inhibitor donepezil in male and both intact and OVX female rats after TBI to determine if motor and acquisition of spatial learning and memory can be facilitated in either group via an additive or synergistic effect of donepezil+EE. Lastly, Aim 5 will expand on the rehabilitative strategy of Aim 4 by determining whether the effects of donepezil+EE are sustained after the subjects are returned to standard living conditions with and without continued pharmacotherapy. Our long-term goal is to develop therapies that facilitate functional recovery after human TBI.

DESCRIPTION (provided by applicant): Approximately 1.5 to 2 million individuals suffer a traumatic brain injury (TBI) each year in the United States and many (24-96%) exhibit agitation and aggression. Antipsychotic drugs (APDs) are commonly used to clinically manage these behavioral sequelae despite a paucity of research into their effect on subsequent recovery. Studies from our laboratory have shown that a short and consistent exposure paradigm (i.e., once daily administration for 19 days) of the APDs haloperidol (HAL) and risperidone (RISP) negatively impact motor and cognitive function after TBI in rats. These detrimental effects may be mediated, in part, by D2 receptor antagonism as parallel experiments using the D2 receptor agonist, bromocriptine, demonstrated enhanced functional recovery. Hence, we propose to empirically evaluate functional recovery after TBI using clinically relevant APDs with different pharmacological actions on the D2 receptor; specifically, HAL (a strong D2 antagonist), RISP (a moderate D2 antagonist) and aripiprazole (ARIP) a partial agonist for the D2 and 5-HT1A receptors. Overall, we hypothesize that ARIP (1) will not be detrimental to functional recovery (because of a lack of D2 receptor antagonist effects), and (2) will enhance recovery (due to D2 and/or 5-HT1A agonist activity). Three specific aims are proposed to test the overall hypothesis. Aim 1 will compare both the short- term (3 weeks) and long-term (3 and 6 months) effects of short-and-consistent exposure of ARIP to HAL and RISP on motor (beam-balance/walk and rotarod), cognitive (spatial learning/memory retention), histological (CA1/CA3 cell survival, cortical lesion volume), Western blot (D2/5-HT1A receptor expression), and TBARS (oxidative stress) outcome after controlled cortical impact (CCI) injury. Aim 2 will compare both the short-term and long-term effects of intermittent dosing (i.e., once every 2, 3, 4, or 5 days) of HAL, RISP, and ARIP on the same endpoints as Aim 1. The rationale for intermittent dosing is that during clinical rehabilitation, not all patients with agitation require daily APD intervention and thus receive prn (i.e., as needed) or intermittent doses. Aim 3 will evaluate the moderating effect of a rehabilitation-relevant environmental enrichment (EE) paradigm on the motor, cognitive, and histological, and oxidative effects of HAL, RISP, and ARIP, examining both short and consistent exposure and intermittent dosing. The EE paradigm mimics the cognitive, physical, and social environment of rehabilitation settings. These various aims are designed to duplicate real world rehabilitation practice in an animal model. Additionally, these aims will inform our understanding of the mechanisms of APDs mediating the deleterious (e.g., HAL and RISP) or potentially beneficial (e.g., ARIP) effects after TBI and could possibly facilitate the discovery o agents to (1) combat TBI- induced agitation and aggression without negatively impacting behavioral recovery, and (2) positively impact outcome by enhancing the recovery process. Furthermore, these studies will have an immediate impact on clinical care by providing clinicians critical information about the effects of APDs after TBI.

Traumatic brain injury (TBI) affects more than 10 million individuals worldwide each year (~ 2.8 million in the USA) and results in long-term motor and cognitive deficits (e.g., reference learning and executive function). To combat this significant health care issue a variety of relatively invasive experimental therapeutic strategies have been attempted and have yielded limited translation to the clinic. Environmental enrichment (EE) is a non-invasive paradigm that promotes significant cognitive recovery and histological protection after experimental TBI and has the potential to mimic post-TBI clinical rehabilitation. The parent R01 was crafted to begin refining and optimizing EE after TBI so that it conformed temporally to clinical neurorehabilitation. The wealth of data lead to a preclinical model of neurorehabilitation that is temporally like the clinic in the sense that delaying EE for a week (i.e., rehabilitation) and providing only 4-hr per day (as common in the clinic) shows significant benefits. Overall, the findings provided significant support for EE as a potential model of neurorehabilitation, but additional empirical research is essential to learn more about its capabilities and limitations that ultimately strengthen its validity and applicability. Hence, the goal of this renewal is to utilize our delayed (7 day) and abbreviated (4 h day) EE model of neurorehabilitation, which we refer to as Rehab, to address questions that concern physiatrists. Five specific aims that are logical and crucial extensions of the parent grant are proposed: Aim 1a determine whether motor (beam and rotarod), cognitive (spatial learning & memory, and executive function using the attentional set shifting test that is analogous to the clinical Wisconsin card sorting task), and affective (open field test) benefits can be sustained after EE is withdrawn, and if so, for how long, Aim 1b determine if providing ?refresher rehab? after the EE-induced benefits begin to wane will stabilize or re-strengthen benefits, Aims 2abc determine whether ?bridging? delayed EE, which is initiated at 7- days after TBI, with a) [amantadine {10 mg/kg/day; i.p.}], b) aqua therapy [{two 90 s swim sessions}], or c) music exposure [(3 h per night of New Age, Ambient, or Classical - Mozart?s sonata for two pianos, K.448)] as adjunct therapies during the week after TBI will augment recovery relative to non-enriched or Rehab groups, and Aim 3 evaluate mechanisms for the bridge plus Rehab therapies. Completion of the aims will further advance a model of neurorehabilitation that mimics the real-world while addressing questions that continue to concern physiatrists, such as how long do the rehab benefits last once discontinued and can they be maintained or improved further with supplemental rehab? Can supplemental therapies before full rehab provide a better outcome? What mechanisms are involved in the effects observed? The refined model will significantly impact and advance rehabilitation-based research. Translatability of the findings will be facilitated further by optimizing the EE model in adult rats of both sexes and by assessing behavior with sensitive and clinically-relevant tests of motor and executive function both acutely and long-term.

The most common neuropsychiatric consequence of traumatic brain injury (TBI) is depression. Early stress exposure has been recognized as an important mechanism for neuropsychiatric disorders in adulthood. In rodents, as in humans, adolescence is a transitional period between child- and adult-hood that is marked by behavioral changes, heightened brain development, and cognitive maturation. Therefore, exposure to adverse environmental conditions during this sensitive period of development could influence TBI psychiatric outcomes. Activation of the endocannabinoid CB1 receptor is thought to suppress the neuroendocrine and behavioral stress response. Although the endocannabinoid system mediates the neuroendocrine stress system, it is unknown how they interact in the pathophysiology underlying emotional behavioral impairments post TBI, or how prior environmental conditions influence these systems, and whether these effects persist in adulthood. The overarching aims are to test the hypothesis that chronic unpredictable stress provided during adolescence will result in deleterious effects on anxiety, depression, and cognition in rats subjected to a TBI as adults. Moreover, the endocannabinoid system may underlie the molecular mechanisms involved in these behavioral changes, and thus treatment with the CB1 agonist ACEA will reduce the deleterious effects of adolescent stress in adults sustaining a TBI. Specifically, the aims are designed to evaluate the long-term effects of chronic unpredictable stress provided during adolescence in adult TBI rats by utilizing a range of tests for social- anxiety- and depression-related behaviors (Aim 1a), and determine whether these behaviors correlate with cognitive impairments, neuroendocrine responses to stress, and levels of markers indicating neuroplastic changes in brain structures mediating emotional and cognitive processes (Aim 1b). Aim 2 will investigate the influence of agonists (ACEA) and antagonists (AM251) of the CB1 cannabinoid receptor on behavioral and neuroendocrine responses (from Aims 1a,b) post adult TBI. Thus, the proposed research will determine whether exposure to repeated stress during adolescence plays a determinant role in modulating components of the stress system and brain plasticity, as well as behavior following adult TBI, and whether such effects are dependent on the endocannabinoid action in brain regions regulating emotional responses. Understanding the impact of environmental and biochemical factors will help develop effective preventive and therapeutic strategies for TBI patients.

Project summary Traumatic brain injury (TBI) affects ten million people worldwide each year. Of those, two million reside in the United States (U.S.). Spinal cord injury (SCI) affects 500,000 worldwide. These CNS injuries are referred to as Neurotrauma and are major causes of disability. While the affective toll of TBI and SCI on interpersonal relationships is incalculable, the financial costs are astronomical. In the U.S. alone, the economic burden of TBI exceeds $75 billion per year. The National Neurotrauma Society (NNS) and the International Neurotrauma Society (INTS) symposia are the primary forums for exchanging the latest discoveries in the fields of TBI and SCI. The NNS, currently in its 36th year, is joining forces with the INTS and the AANS/CNS Joint Section on Neurotrauma and Critical Care (AANS/CNS JSNTCC) for the 3rd Joint Symposium held in Toronto, Canada, August 11-16, 2018. The program planning committee consists of members from each society to ensure a sufficient mix of topics targeting both TBI and SCI, basic science and clinical research, and representative speakers. The joint arrangement successfully brings together basic scientists, clinicians, and trainees from both disciplines from all over the world to network, share concerns, and create solutions for TBI and SCI. The last joint symposium was in 2009 and thus this long overdue meeting promises to be stimulating and filled with exciting state-of-the-art sessions on the consequences of CNS damage and treatment strategies for protection and repair. 7 plenaries and 18 breakout sessions distributed evenly among TBI, SCI, and TBI/SCI are scheduled. Sample sessions include discussions on progress and potential breakthroughs, how stress in TBI and SCI affect recovery, advances in CNS regeneration, and challenges of managing neurotrauma in developing countries. A patient?s perspective segment will afford personal viewpoints on living with injury and thoughts regarding scientific progress in Neurotrauma. Invited speakers not affiliated with neurotrauma, but who are experts in their respective fields will provide novel perspectives. This application seeks funding to support 30 pre-and-post- doctoral trainees to present their ongoing work, attend scientific sessions, and to network with peers and faculty. Of the 30 trainees, 10 will be under-represented minorities (URMs) and persons with disabilities as the societies strive to further promote and retain diversity. To this end, one of the topics of the inaugural Training, Education, and Mentoring (TEAM) organization, which evolved from Women in Neurotrauma Research (WiNTR) to further enhance diversity and inclusivity, will host a presentation (open to all) by Dr. Michelle Jones-London, Chief, Office of Programs to Enhance Neuroscience Workforce Diversity. The program content, which was crafted in response to comments/suggestions from previous meeting attendees conforms to the theme ?Translating Research, Transforming Lives.?

Traumatic brain injury (TBI) affects more than 10 million individuals worldwide each year and results in long- term motor, cognitive, and affective deficits. Pharmacologic strategies are often used to treat TBI but to date no therapy has successfully translated to the clinic, which advocates for other rehabilitative strategies to restore neuronal networks and recover behavioral deficits thereby increasing the probability of bench-to-bedside success. Neural stem cell (NSC) therapies may be a feasible alternative to pharmacotherapies for improving function after TBI. NSC-based therapies can exploit their inherent ability to migrate to stimulate regeneration and repair damaged brain tissue. In our pilot studies, well-characterized allogeneic human NSCs, LM-NSC008, genetically modified to express the human L-Myc gene were intranasally (IN) administered to adult male and female rats after cortical impact injury. LM-NSC008 cells migrated toward and distributed throughout damaged brain tissue and into distant regions mediating behavioral changes. LM-NSC008 cells significantly improved two distinct cognitive domains - spatial learning (reference learning) and executive function vs. vehicle (VEH). Because clinical translation has been unsuccessful with single therapies, the NIH?s TBI and combination therapy workshop recommended the evaluation of combination treatments. We have reported synergistic benefits when environmental enrichment (EE) is combined with pharmacotherapies and predict augmented benefits with LM-NSC008 cells as well. Our hypotheses are that IN LM-NSC008 cells in male and female rats will 1) migrate and accumulate in sufficient quantities at proximal and distal TBI sites and contribute to behavioral recovery, 2) provide benefit with a clinically relevant delayed administration approach, and 3) improve recovery more robustly when combined with EE than when administered alone. To test our hypotheses, optimize IN delivery doses of LM-NSC008 cells, and to determine LM-NSC008 cell fate and mechanisms, alone and in combination with EE, the following Aims are proposed. Aim 1a: Determine the optimal dose and delivery protocol of IN LM-NSC008 cells for maximal distribution to areas of damage at early, delayed, and chronic time points after TBI. A single high dose of LM-NSC008 cells [6x106] or VEH will be given IN on day-7 (acute period), day-21 (delayed), or day-90 (chronic) after moderate TBI or sham injury, while six lower doses [1x106] will be given once on post-surgery days 7,9,11,13,15,17 (acute), 21,23,25,27,29,31 (delayed), or 90,92,94,96,98,100 (chronic) to determine the protocol that provides maximal distribution of cells at the trauma sites at 3 timepoints after TBI and significantly improves recovery. Aim 1b: Evaluate motor, cognitive, and affective behavioral improvements with IN LM-NSC008 cell therapy in TBI and sham rats. Aim 2: Determine the effect of combining IN LM-NSC008 cell therapy with EE on motor, cognitive, and affective behavior. Aim 3: Determine the fate, mechanisms, and regenerative capacity of IN administered LM-NSC008 cells alone or with EE after TBI.