Washington University

NIH Research Projects · FY 2025 · 2021-01

Motor axon loss is a cardinal symptom of amyotrophic lateral sclerosis (ALS). Axon loss can be driven by a genetically encoded program in which the axon survival factors NMNAT2 and STMN2 inhibit the activity of the axon destruction factor SARM1. Recent data suggest that this program of axon self-destruction may contribute to pathology in ALS. First, aggregation of TDP-43, a hallmark of most ALS cases, results in the selective loss of mRNA encoding functional STMN2, a key axon survival factor. Second, loss of SARM1 suppresses some neurodegenerative phenotypes in a mouse ALS model that expresses pathogenic human TDP-43. Here we investigate the contribution of this axon degeneration pathway to ALS. We have defined the mechanism of action of SARM1, demonstrating that it is the founding member of a new class of NAD-cleaving enzymes. SARM1 enzyme activity is normally held in check via an autoinhibitory domain. Injury- or disease- induced loss of NMNAT2 and STMN2 disinhibits SARM1, leading to rapid NAD+ depletion, metabolic catastrophe, and axon fragmentation. Our structure-function studies of the SARM1 protein have identified mutations with a range of consequences, from constitutively active variants that promote cell death and axon loss, to dominant negative variants that are neuroprotective. These findings imply that human variants may exist that either promote or protect against neurodegeneration, and that understanding the phenotypic consequences of genetic variation requires functional studies of enzyme activity. In support of this hypothesis, we have identified several rare SARM1 variants in ALS patients, but not in controls, that have constitutive NADase activity and promote neuron death and axon loss. These variants also cause motor dysfunction and paralysis when expressed in the mouse CNS, suggesting that activating SARM1 mutations may contribute to ALS pathogenesis. Here we propose to define the function of SARM1 variants from ALS patients, controls, and the general population. These studies will allow us to categorize SARM1 variants as putatively pro-degenerative, neuroprotective, or neutral. In parallel, we will dissect the contribution of variation in components of the programmed axon destruction pathway to ALS phenotypes, alone and in combination with known ALS genetic risk-factors, in motor neurons differentiated from human induced pluripotent stem cells (iPSCs). Finally, we will investigate neurodegeneration in a mouse knock-in model carrying a Sarm1 allele equivalent to a pro- degenerative allele found in ALS patients, alone and in combination with a SOD1 model, based on a specific patient genotype that we identified. We will attempt to suppress ALS phenotypes with SARM1 inhibition via a proven gene therapy approach and with experimental small molecule inhibitors. Results of these studies will establish the relationship between the SARM1-mediated axon destruction program and ALS, and build the foundation to develop axoprotective therapeutics to treat this devastating disease.

NIH Research Projects · FY 2025 · 2021-01

Project Summary/Abstract The neural mechanisms of regulation support socioemotional competence 63,74 and may operate as resilience factors in the face of developmental risk for psychopathology 13,17. Investigating how these neural mechanisms emerge and change during infancy may identify critical periods of intervention and individual and contextual factors that condition neurodevelopmental trajectories. Three major gaps exist in this literature: 1) Neural correlates of regulation are well established in the adult literature but have not been examined longitudinally during infancy. 2) The effects of temperament risk, parental psychopathology, and parent-child transactions have only been linked to cross-sectional studies of neural activity, rather than to neurodevelopmental trajectories. 3) The neurocognitive mechanisms underlying the socialization of regulation and psychopathology in parent-child transactions are poorly understood. In this proposal, the first two gaps will be addressed by the dissertation project, and a plan is presented to address the third gap with the postdoctoral research. In the F99 phase, four repeated measures of infant electroencephalogram (EEG) from 8 to 24 months postpartum will be used to map infant trajectories of neural markers implicated in regulation during childhood and adulthood. The EEG data will provide two neural correlates: delta-beta synchrony and network connectivity. At each laboratory visit, infant resting-state EEG is collected, and mother-child interactions are videotaped during a 5-minute free-play task. Additionally, primary caregivers report on their child’s temperament and their own anxiety at each wave. These repeated measures design will enable the applicant to address three main questions: 1) How local and global neural mechanisms underlying regulation emerge and change during infancy. 2) How these neural trajectories are influenced by infant negative affect and maternal anxiety. 3) How dyadic patterns of positive affect and responsivity in the mother-infant relationship impact these neural trajectories. In the K00 phase, the applicant will use novel techniques to assess dyadic neural and cognitive activity in mother- child interactions in order to map specific regulation socialization strategies to neural synchrony, joint attention, and risk for anxiety. The current proposal entails an integrated pre- and postdoctoral training plan to prepare the applicant as an independent researcher in developmental neuroscience. There are four overarching training goals: 1) Gain expertise in the study of infant brain development. 2) Learn and apply advanced quantitative methods and dyadic analysis to the study of brain development. 3) Expand dyadic analysis of mother-child interactions to the neural and cognitive systems and. 4) Develop strong scientific writing and teaching skills. These training goals will effectively prepare the applicant to transition from the predoctoral to the postdoctoral training and build a career program that can be taken into an independent, tenure-track position as a developmental neuroscientist.

NIH Research Projects · FY 2025 · 2020-12

Use of the non-dominant left hand is critical for individuals who suffer chronic impairment of the dominant right hand due to unilateral conditions such as peripheral nerve injury. (This project will study right-handed individuals, and thus uses "right hand" instead of "dominant hand.") Most rehabilitation focuses on restoration of function, but many patients never achieve this: 64,000 people per year in the USA have nerve injury to the right hand but achieve satisfactory recovery, and these patients must learn to compensate by using the left hand. However, the neural mechanisms of left hand compensation remain unknown. Our preliminary data suggest that compensation involves interhemispheric mechanisms: the left hemisphere's mechanisms are recruited to support the ipsilateral left hand. However, this mechanism has never been assessed with neuroimaging during the left hand precision movements that would engage such a mechanism, nor in the context of hand usage choices (left vs. right) during unconstrained reach-to-grasp action. Our short-term goal is to identify interhemispheric mechanisms that support left hand compensation (both performance and use), and determine whether the mechanisms arise from cortical asymmetry for movement (hand dominance). This will provide the foundation for our long-term goal to develop and target therapies to improve compensation for patients who face challenges to rehabilitation due to chronic right hand impairment. Our patients will be individuals with chronic forced use of the left hand due to unilateral upper extremity peripheral nerve injury. We will compare them with healthy patients in one fMRI study with 3 Aims: Aim 1: identify interhemispheric mechanisms that support left hand performance after right hand injury. We expect left hemisphere activity to correlate with left hand performance in fMRI, in patients > controls. Aim 2: identify interhemispheric mechanisms that support left hand usage after right hand injury. We expect left hemisphere activity to correlate with left hand usage outside fMRI, in patients > controls. Aim 3: determine whether the interhemispheric mechanisms arise from cortical asymmetry. We expect the mechanism to depend on hemisphere-specific specializations. Specifically, for patients who retain some function of their injured right hand, we expect that ipsilateral brain involvement will be demand-correlated during left hand action > during right hand action. These findings will establish a mechanistic understanding of the interhemispheric cortical mechanisms of left hand compensation. These mechanisms are the necessary foundation for future development of interventions such as targeted neuromodulation, and precision-medicine prediction of which patients will benefit from compensatory therapy. Moreover, our findings will establish a healthy-brain mechanism for the changes following chronic forced use of the LH, which can serve as a baseline for future studies of central nervous system conditions (e.g. stroke) that include chronic forced use.

NIH Research Projects · FY 2025 · 2020-12

PROJECT SUMMARY/ABSTRACT Deficits in effortful control (EC; top-down control of self-regulation involving inhibitory control and executive attention) represent a major transdiagnostic risk factor for multiple psychiatric disorders in childhood. Dysfunctional parenting is a strong predictor of impaired EC, making it a potent target for intervention. Dysfunctional parenting includes behavioral deficits in parenting sensitivity [timely and appropriate responses to changes in infant physical and emotional needs], along with cognitive deficits in maternal mind-mindedness [MMM; attunement to infant mental states that govern goal-directed behavior]. MMM may underlie parenting sensitivity, but prospective links remain unclear. MMM facilitates the transition from external caregiver-based regulation to self-regulation in early childhood by providing the foundation for intentional mental-state talk to regulate behavior. While adverse caregiving exposures have profound effects on brain structure, the extent that MMM impacts infant brain structural and functional connectivity in key brain regions underlying EC during periods of brain plasticity is unknown. This innovative proposal offers an unprecedented opportunity to address this scientific question with training at an institution rich with expertise in child psychiatry and neuroscience research. This K01 will chart the development of EC and related brain networks by leveraging a valuable captured infant cohort undergoing repeated diffusion (dMRI) and resting state functional (rsfMRI) MRI scans and developmental evaluations from birth to age 3 years. The applicant will add to the existing study a prospective observational assessment of MMM, a standardized test of infant executive attention, and examine connectivity in a unique set of white matter tracts. As the cohort is enriched for poverty, this study will examine novel links between maternal perinatal psychosocial risk (poverty, depression, anxiety, stress) and later MMM and parenting sensitivity, and the extent that that psychosocial risk indirectly influences EC via causally linked multiple mediators of MMM and infant brain connectivity. Findings will elucidate MMM as a modifiable factor that may increase parent sensitivity to enhance infant brain connectivity and EC to reduce psychopathology. [If MMM alters infant brain connectivity, findings will underscore infancy as a critical time for targeted preventative intervention before EC deficits emerge and lead to psychopathology; identify vulnerable infants who may benefit the most from intervention; and inform the design of novel MMM interventions to enhance infant brain function in networks most sensitive to parenting inputs and most related to EC.] This award will build upon the applicant's foundation in child development, school- age executive attention, and structural MRI by providing [new and more advanced training in the assessment of mother-infant interactions, the development of EC in infancy, and dMRI and rsfMRI methods.] The applicant will gain the skills needed to conduct longitudinal research on the caregiving and neurobiological mechanisms of child psychopathology. This K01 will facilitate an R01 proposal that will modify MMM in a parent-child intervention to alter risk trajectories linking poverty with aberrant brain connectivity and psychopathology into school-age.

NIH Research Projects · FY 2025 · 2020-12

PROJECT SUMMARY Multiple sclerosis (MS) is an inflammatory demyelinating disease with, ultimately, irreversible axonal injury leading to permanent neurological disabilities. Preventing disease progression or treating progressive MS remains a major unmet clinical need. We have previously developed a novel data-driven model-selection diffusion basis spectrum imaging (DBSI) to accurately image inflammation, demyelination, and axonal injury, as well as quantifying axonal loss in the presence of vasogenic edema in experimental autoimmune encephalomyelitis (EAE) and spinal cord injury mice, and brain WM pathologies in MS. MRI does not distinguish inter- from intra-axonal water signals, reflecting a weighted-average of signals between the two compartments. However, our recent observation that DBSI derived axial diffusivity (DBSI-λǁ) was slightly elevated in normal appearing white matter (NAWM) in people with MS (pwMS). This elevated DBSI-λǁ added uncertainty in assessing whether axonal injury (against the notion that ↓DBSI-λǁ ≈ axonal injury) is present in NAWM of these pwMS. In this proposed study, we will refine DBSI to further improve its sensitivity and specificity to axonal injury/loss, demyelination, and inflammation for accurately assessing disease progression and therapeutic efficacy in pwMS. Since MRI does not distinguish inter- from intra-axonal water signals, it reflects a weighted-average between inter- and intra-axonal signals. In the presence of inflammation-associated edema or minor axonal loss in pwMS, the longer diffusion time for human scanners coupled with the increased inter-axonal space will lead to increased DBSI-λǁ masking the detectability of axonal injury. Thus, through separating inter- and intra-axonal water compartment signals, the sensitivity and specificity to axonal injury of DBSI-derived intra-axonal λ|| (DBSI-IA-λ||) may be improved. This new model will still preserve the isotropic diffusion specificity to inflammation and tissue loss. We propose three specific aims to prove or disprove this hypothesis: Aim 1. To perform DBSI and DBSI-IA analyses on autopsy specimens from pwMS followed by conventional histology and immunohistochemical staining. Aim 2. To perform DBSI and DBSI-IA modeling on perfused frog sciatic nerve with and without contrast agent to separate inter-/intra-axonal space water signal. Aim 3a. To develop a Diffusion Histology Imaging (DHI) approach combining DBSI/DBSI-IA metrics and machine/deep learning algorithms to recapitulate histology specificity to MS pathology. Aim 3b. To translate DBSI-IA model to analyze existing DWI data from the cohort of pwMS previously imaged in an expired program project.

NIH Research Projects · FY 2025 · 2020-12

Recent advances in human genetics have defined hundreds of causal variants for Autism Spectrum Disorder (ASD) and other intellectual and developmental disabilities (IDDs). However, substantial effort is required to define downstream disease processes and thus to guide development of interventions for each one. One such new ASD gene is MYT1L – while mutations in MYT1L have recently become associated with ASD and Intellectual Disability (IDD) in humans, the role of MYT1L in neural cells and circuits is unclear. Thus, here, we propose a comprehensive mechanistic investigation of MYT1L. This project uses both cutting-edge established workflows as well as innovative new approaches to enable in-depth study of MYT1L loss at the molecular, cellular, structural, and behavioral circuit levels. We will utilize two complementary experimental systems, mouse models and human induced pluripotent stem cell (iPSC)-derived neurons, to define MYT1L's normal roles, and to identify the consequences and reversibility of MYT1L loss. We focus initially on a mutation identified in a patient with a MYT1L putative loss-of-function variant who has ASD and ID. In addition to knock-in of this variant into isogenic control PSC lines, we derived iPSC models from this subject, with and without MYT1L variant correction, enabling us to define consistent consequences of MYT1L mutation across human genetic backgrounds. We also developed mouse models targeting the paralogous amino acid, to enable studies of the consequence of MYT1L loss on brain structure, physiology, and behavioral circuit function. Further, cutting-edge gene therapy-like tools developed for both mouse and human models will allow us to investigate the effects of rescuing gene function. Similar landmark experiments profoundly changed the understanding of other neurodevelopmental disorders by demonstrating that a substantial proportion of the phenotype was reversible, thus spurring the development of therapeutics based on rescuing gene expression. Together, the experiments performed here will elucidate the requirements for and mechanisms by which MYT1L controls brain development and function, will determine how these are disrupted by pathogenic MYT1L mutation, and could also chart a course towards MYT1L-targeted therapies.

NIH Research Projects · FY 2025 · 2020-12

Abstract. UBE3A is a gene that encodes a HECT (Homologous to E6AP C-terminus) domain E3 ubiquitin ligase linked to numerous developmental and psychiatric disorders. Loss of function of the maternally derived UBE3A enzyme causes Angelman syndrome whereas gain of function, due to duplication or triplication of UBE3A, is linked to a broad range of disorders including Dup15q syndrome, schizophrenia, and mood disorders. These observations strongly suggest that bi-directional changes in UBE3A activity contribute to neuropsychiatric pathology. However, the mechanisms that regulate UBE3A activity remain poorly understood. The primary goal of our proposed research is to uncover mechanisms of UBE3A regulation, that when faulty, can lead to aberrant gain or loss of UBE3A function. In preliminary work, we developed a high-throughput assay to assess the functional consequence of non-truncating UBE3A missense variants. This screen identified numerous novel loss of function mutations, as well as gain of function mutations that hyperactivate UBE3A activity well above wild type (WT) enzyme levels. These results provide deep structure-function information that we can now leverage to uncover mechanisms of UBE3A regulation. This proposal aims to, 1) create a complete functional catalogue of known missense variants identified in individuals, 2) utilize structure-function analyses to identify mechanisms that can lead to both aberrant gain and loss of enzyme function, and 3) leverage biochemical insights to engineer proteins that can target UBE3A activity. The molecules generated from our work will be applied to examine whether alteration of UBE3A activity can rescue synaptic phenotypes observed in mice harboring hyperactivating mutations in UBE3A. If successful, our work will provide new biochemical insights and tools that will make it possible to target UBE3A for therapeutic intervention in various neuropsychiatric disorders.

NIH Research Projects · FY 2025 · 2020-12

PROJECT SUMMARY Glioblastoma (GBM) is a brain tumor that causes neurological deterioration and death in most patients within 2 years. Despite aggressive therapy, most GBMs reappear within 6 months. A major reason for this outcome is because GBM stem cells (GSCs) are resistant to existing therapies. Currently, no treatment consistently kills these highly resistant cells. However, the Zika virus epidemic has provided us with a new approach to eradicate GSCs. ZIKV targets normal stem cells in the developing fetal brain, yet has minimal effects on differentiated neurons or the adult brain. Since GSCs share properties with neural stem cells, we investigated whether the natural honing and lytic activity of ZIKV could be harnessed to target and kill GSCs. We published the first use of ZIKV to kill GSCs. We showed that ZIKV kills GSCs in tumors removed from patients, with minimal impact on non-GSC tumor cells, called differentiated GBM cells. Importantly, normal human brain cells were not affected by ZIKV. After intracranial treatment with ZIKV, mice harboring gliomas survived more than twice as long as untreated mice and, in some cases, treated mice were long-term survivors. In addition to the resistance of GBM stem cell to chemoradiation, GBM is the hallmark example of an immunotherapy-resistant tumor. Importantly, we found that in vivo, ZIKV treatment reduces tumor size and extends survival beyond that expected for only anti-GSC effects. This suggested that ZIKV killing of GSCs may trigger an immune response against the remainder of the tumor. In more recent studies, we have found that CD8+ T cells are required for the efficacy of ZIKV as an oncolytic therapy in vivo. Our central hypothesis is that ZIKV elicits an anti-tumor immune response that could be made even more effective by combining it with existing immunotherapies. Aim 1 will determine how CD8+ T cells promote tumor clearance after ZIKV and Aim 2 will determine whether ZIKV can be combined with immunotherapy or standard-of-care for GBM to improve outcomes. Our long-term goal is to develop a new treatment for GBM by leveraging the immune system response to ZIKV.

NIH Research Projects · FY 2024 · 2020-09

Project Summary/Abstract Sleep is necessary for all brain function and ultimately life. The core function by which sleep contributes to healthy cognition remains one of the great questions facing neuroscience. Recent theories point to powerful cellular rules, but these are controversial and have difficulty accounting for the effects of sleep in ethologically and developmentally diverse circumstances. We recently showed that cortical circuit dynamics are actively tuned to criticality, a computational regime that maximizes information processing. This regime is disrupted by changes in synaptic strength, such as those believed to typify waking experience. This raises the intriguing possibility that the core mechanism by which sleep benefits neural function is by restoring criticality. Our preliminary experiments support this hypothesis. The overall goal of this project is to develop a new framework for understanding the neural impact of sleep and experience at the level of network dynamics. We will continuously track 500-1000 single neurons in the brains of freely behaving animals for up to six months. We will track sleep, wake, behavior, and neural dynamics across the entire distribution of naturally occurring behavior. We will take advantage of this methodologically integrated approach to understand how circuits in the brain maintain the stable computation necessary for cognition and natural behavior on long time-scales. In Aim 1, we will test the relationship between specific classes of behavior and criticality in underlying networks. In Aim 2, we will test the impact of wake and sleep on criticality across the brain for the majority of an animal's lifetime. In Aim 3, we will use a modelling-based approach to establish the mechanisms of criticality in the intact brain. The results of this work will shed light on a long-standing gap in our knowledge of fundamental neurobiology. Given the increasingly recognized role of sleep in a vast number of brain-related disorders, an understanding of how sleep works will open the door to significant health-related progress in the future. This work directly advances the mission of the BRAIN Initiative.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY/ABSTRACT Globally, there were an estimated 36.9 million people living with HIV in 2017. As a result of the increased availability of antiretroviral therapy, life expectancy for these patients has increased, resulting in an expansion and aging of the HIV-infected population. Data from both high- and low-resource countries have shown an escalating burden of heart, lung, blood, and sleep (HLBS) disorders among HIV patients. Care for this now aging cohort requires modifying service delivery models towards the prevention, diagnosis, treatment, and control of chronic HLBS disorders. The successful implementation of various solutions developed in response to the global HIV/AIDS pandemic can serve as the foundation to transform global healthcare delivery systems towards HLBS disorder care. The overarching goals of this Research Coordinating Center are: (1) provide scientific, organizational, statistical, quality control, capacity building and operational leadership for the Implementation Science projects to be funded by the companion RFA-HL-20-025 and (2) promote and teach Implementation Research strategies to advance collaborative dialogue across projects and ensure up-to-date conceptualization, specification and measurement of implementation strategies and outcomes. In support of these goals, we have assembled a highly experienced and multidisciplinary research team. Our team, with expertise in running multicenter trials, dissemination and implementation research, biostatistics and research design, HIV, HLBS disorders, capacity building, and data management, will pursue the following specific aims: 1. To oversee management, organizational, and communication activities across all research sites including working with NHLBI to prepare meeting agendas, develop the study website, provide administrative support for research projects, and organize face-to-face meetings. 2. To collaborate with all research sites in developing the scientific and operational details of their implementation science research protocols. These efforts will include: a. Assisting study sites in designing, testing, and maintaining password protected data entry and management systems and ensuring that such systems implement robust quality control features. b. Harmonizing (a) demographic, clinical, and questionnaire measurements across research sites and (b) the conceptualization and measurement of implementation strategies and implementation outcomes. c. In collaboration with research sites, to statistically analyze harmonized data and to assist in the analysis of site-specific data as reports, manuscripts, and abstracts are prepared. 3. To measure critical contextual Implementation Science factors in each research setting in order to enable conceptualization of the mechanism of effect and, by extension, enable formalized assessments of external validity. To oversee the development and implementation of training and capacity building activities.

NIH Research Projects · FY 2024 · 2020-09

Project Summary/Abstract Making sense of visual scenes requires a correct assignment of the borders that occur between object and background, or between objects, to the foreground object. Macaque visual cortical areas V2, V4 and V1 contain neurons that are selective for border ownership. These cells encode border ownership even if distal visual cues that define which sides of a boundary are object and ground fall far outside of the neuron’s receptive field. This selectivity often persists even when these distal cues disappear, a form of stimulus hysteresis or memory. Prior studies suggest that this border ownership selectivity relies on corticocortical feedback from hypothetical downstream neurons with receptive fields that cover the complete object, termed grouping cells. No prior study has found these hypothesized grouping cells. Thus, though border ownership cells have been found, the neural circuits that endow them with border ownership selectivity remain poorly understood, as does their role in perception. The goals of the proposed research are to determine the micro-organization of border ownership and grouping cells in the macaque visual cortex, and to relate their activity to perceived border ownership. The candidate will use advanced electrophysiological, optophysiological and viral targeting techniques in behaving macaques to achieve these goals. In Aim 1, the candidate will use two-photon calcium imaging to identify border ownership cells and grouping cells, and test specific hypotheses about how they are organized within the columnar layout of macaque Area V4. In Aim 2, the candidate will combine two-photon calcium imaging and viral targeting techniques to distinguish excitatory from inhibitory neurons in V4, and assess their role in border ownership. In Aim 3, the candidate will relate the activity of border ownership and grouping cells in areas V2/V1 and V4 to perception by recording their activity with laminar multielectrodes while reading out perceived border ownership. The candidate has extensive electrophysiological expertise including in behaving non-human primates, but needs training in two-photon calcium imaging and viral approaches, which are the technical goals of the career development plan. The primary mentor is Dr. John Reynolds, a leader in the neurophysiology of visual cortex in behaving macaques. The co-mentor is Dr. Ed Callaway, a pioneer in viral targeting and two- photon imaging in macaques. Both Dr. Reynolds and Dr. Callaway have a strong history of mentoring young scientists and are faculty at the Salk Institute for Biological Studies, an institute with a strong history in visual neuroscience. Recent advances in optical recording techniques and viral approaches have enabled high- resolution studies of genetically targeted neurons in functioning neural circuits. Combined with his background in electrophysiology, the additional training will provide the candidate with the expertise that will enable him to launch a successful career as an independent investigator studying the neural basis of visual perception in behaving non-human primates. A deeper understanding of the computations performed in the primate visual cortex is imperative for designing better diagnostic tools and treatments for central visual processing disorders.

NIH Research Projects · FY 2024 · 2020-09

Project Summary Although 21% of pregnant women experience perinatal mood and anxiety disorders, the burden is especially heavy for low-income and minority women. For example, African-American women have a higher risk of psychosocial stress (e.g., exposure to adversity, racism, and traumatic life events) throughout life than white women. Moreover, African-American women are at significantly higher risk for maternal and neonatal adverse events resulting from psychological distress, such as preterm birth (17% vs. 10%) and low birthweight (13% vs. 7%). However, African-American women are the least likely group to receive mental health interventions that could reduce these disparate outcomes. A cross-system collaboration between researchers, clinicians, and patients is working to overcome these barriers by developing, testing, and implementing a novel model: Elevating Voices, Addressing Depression, Toxic Stress and Equity in Group Prenatal Care (EleVATE GC). EleVATE GC is based on group prenatal care and has embedded within it a trauma-informed, evidence-based behavioral health intervention grounded in anti-oppressive principles. The objectives of this proposal are to rigorously assess the effectiveness of EleVATE GC and to determine the feasibility, sustainability, and barriers to implementing EleVATE GC in real-world care settings. These objectives will be achieved by conducting a pragmatic effectiveness-implementation randomized controlled trial comparing EleVATE GC (n=563) to individual prenatal care (n=282) for pregnant women at high risk for depression. This trial will be conducted at eight diverse prenatal clinics in the St. Louis region that serve a population with a high preterm birth rate (~16% vs. 11% nationally). Additionally, 80% of women served by these sites have a history of depression, anxiety, trauma, or another mental health diagnosis. Within this trial, Aim 1 is to determine the effectiveness of EleVATE GC to reduce perinatal depression and adverse pregnancy outcomes among low-income, predominantly African-American women. This aim tests the hypothesis that, compared to women receiving individual care, those in EleVATE GC will have lower perinatal depression (primary) and lower risk of preterm birth and low birthweight infants (secondary). Aim 2 is to identify strategies and contextual implementation factors to enhance implementation of EleVATE GC. An adaptation of the Practical Robust Implementation and Sustainability Model will be used across three of the four domains (intervention design, recipient, implementation and sustainability) at three levels (prenatal care clinic, clinician, and patient). This project is directly responsive to the call of RFA-MH-20-400 to "test the effectiveness of strategies for implementation and sustainable delivery of evidence-based mental health treatments and services to improve mental health outcomes for underserved populations in under-resourced settings in the United States".

NIH Research Projects · FY 2024 · 2020-09

Project Summary This is a study to investigate the relationship between trajectories of alcohol use, longitudinal changes in brain function, and the development of Alzheimer disease (AD). To address gaps in knowledge about the relationship between alcohol use and AD, we will integrate plasma Aβ testing and a measurement of clinical dementia into ongoing assessments of N=600 participants (age ≥ 50, 17% African American) in a large ongoing study of alcohol use disorder. We will leverage sample collection from the St. Louis site of the Collaborative Study on the Genetics of Alcoholism (COGA), a longitudinal, family-based study of alcohol use disorder funded by NIAAA for over 30 years, with extensive clinical, neuropsychological, electrophysiological, and genetic data from families densely affected by alcohol use disorder and community-based comparison families. The ongoing assessments of older COGA participants includes a comprehensive evaluation of alcohol use, neurophysiological measures including resting-state electroencephalogram (EEG) and event-related brain potentials (ERPs) acquired during cognitive tasks (same as in previous longitudinal assessments), and neuropsychological surveys. Together with existing COGA data, the new combined assessment will allow for creation of powerful measures of alcohol use, brain function, and neuropathology. This represents the first study to integrate AD biomarkers with comprehensive, longitudinal assessments of alcohol use. Aim 1 will examine the effect of alcohol consumption on preclinical AD and longitudinal changes in brain function and cognition in older adults. Aim 2 will investigate genetic, comorbid, environmental, and demographic factors as moderating the effect of alcohol consumption on AD biomarkers and brain function. The innovations include integration of state-of-the-art AD assessment, plasma biomarker of AD, brain function measures of neural synchronicity and connectivity, and comprehensive longitudinal assessment of alcohol use in a high-risk sample that has been followed over 20 years. This proposal is significant because the products and results will apply broadly to our understanding of both the development of AD and the long-term impact of alcohol on the brain.

NIH Research Projects · FY 2024 · 2020-09

Project Summary Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a positive-sense single-stranded RNA virus that was first isolated in Wuhan China in December, 2019. SARS-CoV-2 is the cause of coronavirus disease 2019 (COVID-19), which is now a pandemic and has caused more than 1.3 million confirmed cases and 72,000 deaths, with an estimated case fatality rate of 4%, with substantially higher death rates (~15%) in the elderly or immunocompromised. Virtually all countries and territories have reported cases, with major epidemics in China, Italy, Spain, France, Germany, Iran, and the United States. SARS-CoV-2 is thought to be of zoonotic origin, most likely bats, and is about 75% identical to the original SARS-CoV. Most cases are spread by direct human-to- human transmission, with community transmission in asymptomatic individuals described. Currently, no countermeasures are licensed for human use. The development, characterization, and ultimately deployment of an antibody-based treatment against SARS-CoV-2 could prevent substantial morbidity and mortality, and possibly mitigate its epidemic spread. This interactive multi-PI proposal leverages complementary expertise in the Diamond, Crowe, and Baric laboratories to rapidly develop highly neutralizing and therapeutic human monoclonal antibodies (mAbs) against SARS-CoV-2 for immediate use in humans. To achieve this goal, we will generate and interrogate human mAbs against SARS-CoV-2 that are obtained from multiple convalescent subjects. We will identify potently neutralizing mAbs and optimize them for affinity by selecting naturally occurring somatic variants identified by repertoire sequencing and sibling analysis and Fc effector functions. Protective activity of top candidate coronavirus mAbs will be tested in newly-generated and optimized mouse models of SARS-CoV-2 infection, including those expressing human ACE2 receptors (hACE2). To define correlates of protection, we will use chimeric viruses, shotgun mutagenesis, and neutralization escape to identify the epitopes of our most protective mAbs. Our team has extensive experience in the generation, characterization and optimization of antibodies, CoV biology, and animal models of disease and protection. A therapy composed of one to three highly neutralizing mAbs may provide an immediate countermeasure against the pandemic spread of SARS-CoV-2 and help establish correlates of structural and functional humoral protection that ultimately inform vaccine efforts.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY/ABSTRACT: Parkinson disease dementia (PDD) – one of the Alzheimer's Disease Related Dementias (ADRD) – is associated with increased disability, mortality and healthcare costs among people with PD. The long term goal of this research is to reduce disability and delay the onset of the ADRD PDD by enabling people at risk for PDD to cope with cognitive decline to maintain daily function. Existing medical treatments for PD do not prevent or treat cognitive impairment, so behavioral interventions that mitigate its negative functional consequences and potentially delay or slow PDD are a top research priority. Unfortunately, the widely-used process training approach to cognitive intervention (repetitive practice of tasks designed to challenge and improve particular cognitive functions) has been unsuccessful in improving daily function and quality of life in this population. To overcome this limitation, the investigators take a strategy training approach, teaching targeted strategies that people can use in their everyday lives to circumvent cognitive deficits and accomplish daily tasks. Strategy training is a practice standard for cognitive rehabilitation in brain injury and stroke, but its application to PDD is novel. Moreover, the strategy training intervention developed for this study explicitly focuses on generalization of training to daily function, an aspect that is critically lacking from many typical strategy training approaches and from PD-related cognitive intervention research to date. This project addresses prospective memory, or the ability to remember to execute delayed intentions at the appropriate moment in the future. Good prospective memory is essential for independent living, employment, social relationships, and compliance with important health behaviors. People at risk for PDD have prospective memory deficits that relate to worse daily function and quality of life. Therefore, prospective memory impairment is a relevant cognitive problem to address in this population. The primary objective of the current project is to determine the efficacy of mechanistically-targeted strategy training on prospective memory among people at risk for PDD (PD with subjective cognitive decline). It is a single-blind randomized controlled trial comparing the effects of strategy training to the traditional process training approach on objective laboratory prospective memory performance (Aim 1) and reported everyday prospective memory function (Aim 2). Additional objectives are to investigate neural mechanisms of prospective memory impairment in PD (Aim 3) and neural and behavioral predictors of prospective memory training response (Aim 4). This project leverages participants, data and infrastructure from an existing longitudinal cohort of PD and control participants to evaluate short-term and long-term training effects, neurobiological mechanisms, and predictors of treatment response. This work will meet the pressing need for effective cognitive interventions that delay functional impairment associated with PDD and will positively impact function and quality of life in this population.

NIH Research Projects · FY 2025 · 2020-09

PROJECT SUMMARY The DIAN-TU platform was formed to design and manage interventional therapeutic trials and find a treatment that provides cognitive benefit for those certain to develop dominantly inherited AD (DIAD). The DIAN-TU trial platform is now fully operational in 13 countries and 37 sites. The current DIAN-TU secondary prevention trial is a four-year phase 3 cognitive endpoint trial of two anti-amyloid drugs, solanezumab and gantenerumab, with results to be announced in early 2020. The DIAN-TU platform is now mature and primed for testing treatments targeting tau or a combination of tau and amyloid-beta (Aβ) depending on the outcomes of the amyloid trials, and is the ideal platform to provide pivotal biologic results of tau treatment in a pure form of AD. If there is a positive outcome of Aβ drugs, this would support all subjects to be on therapy, enabling combination treatment and giving tau drugs a greater chance of success. If Aβ drugs are negative, then we will need to address the ability of tau-targeted drugs to impact the biology of AD and the potential to slow or prevent the disease. Thus, the tau NexGen studies can and should be done regardless of the outcomes of current amyloid trials. The next phase of the DIAN-TU Next Generation (NexGen) trial will test diverse tau targets in the DIAD population using three mechanisms: a tau antibody, a genetic treatment, and an aggregation inhibitor. The DIAN-TU Tau NexGen will conduct randomized, double blind, pooled placebo-controlled, two-year phase 2 biomarker endpoint trials of three anti-tau or anti-tau/anti-Aβ combination therapies in 216 DIAD mutation carriers (MCs, 72 in each drug arm) who are mildly symptomatic (CDR 0.5 or 1) or asymptomatic with an estimated year of symptom onset (EYO) 15 years before to 10 years after EYO. The trial platform has five novel trial design aspects: 1) a dose escalation algorithm to safely maximize target engagement; 2) a common-close design, so all subjects will stop treatment when the last enrolled subject completes the two- year treatment; 3) a pooled placebo/control design, which increases the number of subjects who contribute to the primary analysis; 4) novel imaging (e.g., the latest generation tau PET tracer, diffusion basis spectrum imaging MRI) and biofluid measures of soluble tau species, neurodegeneration, and inflammation; and 5) a cognitive and tau PET run-in period. The DIAN-TU's contribution is expected to be a substantial understanding of the key tau and combination therapeutic targets in AD through the use of an innovative trial platform in an ideal population. This contribution would provide the AD field with a greater likelihood of translating promising anti-tau therapies to large phase 3 studies, accelerating disease-modifying therapies in DIAD, and possibly translating to the more common sporadic form of AD. Our future aims are to transition successful phase 2 biomarker outcome studies to phase 3 cognitive endpoint outcome studies to enable registration of successful drugs.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY In the proposed work, the Hunstad laboratory at Washington University will employ new models of urinary tract infection (UTI) in female, androgenized female, and male mice to determine molecular mechanisms by which host androgen exposure promotes uropathogenic Escherichia coli (UPEC) pyelonephritis as well as renal scarring, a common sequela of upper-tract UTI. Our recent findings indicate that the influence of sex, including androgen exposure, on these common bacterial infections is more complex than previously appreciated. Using newly developed and optimized models of UTI in mice, we recently demonstrated that androgen exposure is associated with increased risk for chronic cystitis and high-titer pyelonephritis, as well as formation of renal abscesses, in both male and female hosts. These findings correlate with epidemiologic data revealing higher morbidity and mortality in men who do suffer complicated UTI (compared with women), and higher incidence of UTI in women with a common hyperandrogenic condition (polycystic ovary syndrome). Moreover, we have demonstrated a separable effect of androgens on renal fibrosis (scarring), a common complication of pyelonephritis in children that can contribute to long-term sequelae such as hypertension and risk for chronic kidney disease. On the basis of these findings, we hypothesize that fundamental sex differences impact the host-pathogen interaction and cellular responses in pyelonephritis, thereby influencing pathogenicity, resolution, and subsequent renal scarring. To interrogate this hypothesis, and building on our published work, we will first comprehensively define host-sex-specific virulence requirements and sex influences on the host- pathogen interaction through single-nucleus RNA-seq, hybrid capture-enhanced bacterial RNA-seq, and insertion-site sequencing experiments on the infected kidney, in collaboration with MPI Dr. Earl and collaborator Dr. Humphreys. Candidate sex-discrepant host pathways and bacterial virulence factors will be confirmed and interrogated using germline and conditional knockout mice, bacterial mutants, and an array of bacterial pathogenesis studies. We will also define the molecular pathways underlying renal fibrosis following pyelonephritis, and how these pathways are modulated by sex and/or by androgen exposure at varying times before, during, and after UTI. In total, the proposed work will leverage new preclinical models and an array of conventional and cutting-edge experimental techniques. Our results will illuminate sex-specific host-pathogen interactions in the infected kidney and identify UPEC virulence factors important in sex-dependent outcomes of upper-tract UTI. In addition, our preclinical findings will also be translationally relevant to recurrent UTI and renal scarring in human patient populations.

NIH Research Projects · FY 2024 · 2020-09

The islet of Langerhans plays a key role in glucose homeostasis through regulated hormone secretion. Islet research has focused on the insulin-secreting β-cells, even though aberrant secretion of another islet hormone, glucagon from α-cells, exacerbates the pathology of diabetes. Normalization of glucagon action by glucagon receptor antagonism can help diabetic patients maintain euglycemia and avoid hypoglycemic episodes, but progress on this approach has been slowed by numerous side-effects. An alternate approach would be to reduce glucagon secretion, but the mechanism controlling its exocytosis remains controversial. Recent data from our lab and others suggest that the long-standing focus on α-cell Ca2+ signaling may have been misleading, and that regulation of glucagon secretion requires signaling through multiple pathways. This complexity drives an innovative research strategy where we integrate data from mechanistic experiments focused on individual components, while also testing for possible cross-talk between pathways. The loss of glucose-regulation of glucagon secretion (GRGS) in vivo after β-cells are destroyed in Type I diabetes suggests that interactions between islet cell types are critical to α-cell function. This has led to models of paracrine signaling where secreted factors from islet β- and δ-cells constrain α-cell function. We have shown that insulin and somatostatin work in concert to reduce cAMP and PKA activity, which lowers glucagon secretion, but does not by itself explain GRGS. Preliminary data point to a key role for complexin 2 in linking PKA activity to secretion. We have also shown that a novel juxtacrine pathway, EphA4/7 forward signaling, is activated by ephrinA5 ligands on the β-cell surface. This effect leads to F-actin polymerization and decreased glucagon secretion, putatively via RhoA activation, and it appears to have a glucose-regulated component. Thus, both paracrine and juxtracrine pathways drive GRGS from islets, but dispersed α-cells treated with ephrinA5 also exhibit an additional GRGS mechanism, which appears to be intrinsic to the α-cell. Based on these data, we hypothesize that GRGS requires a synergistic combination of paracrine, juxtacrine, and cell-intrinsic signaling pathways. This hypothesis will be tested via three specific aims: 1) Determine the role of paracrine-mediated PKA-activated phosphorylation of complexin 2 in insulin- and somatostatin-mediated inhibition of glucagon secretion; 2) Determine the role of RhoA activation in the juxtacrine EphA4/7 forward signaling that leads to inhibition of glucagon secretion; 3) Determine the role of EphA4/7 forward signaling in intrinsic α-cell response to glucose. The multiple intracellular and intercellular signaling mechanisms that we are uncovering will be elucidated by methods that allow precise observation of the pertinent dynamics in living cells and islets. The research plan will also leverage our findings that the mechanisms of GRGS are similar in mouse and human α-cells, and we will perform parallel experiments across species to the extent possible. These experiments will further our understanding of α-cell function, which is a critical step towards discovering new potential targets for the regulation of glucagon and treatment of diabetes.

NIH Research Projects · FY 2025 · 2020-09

Project Summary/Abstract Advanced chronological age is the greatest risk factor for developing Alzheimer’s disease. Accumulating evidence suggests that disease processes may begin decades prior to dementia. Therefore, cellular and molecular processes that contribute to biological aging may also modulate Alzheimer’s disease pathogenesis. We recently identified a fundamental cellular aging stress response, cellular senescence, as a pathogenic process driving neurodegeneration in tauopathies, brain diseases histologically defined by tau protein accumulation. Features of cellular senescence include stable cell cycle arrest and toxic secretory phenotype. In this way, senescent cells escape cell death and become persistently deleterious to their surrounding environment. We have found a causal relationship connecting tau accumulation (i.e. neurofibrillary tangles), a neuronal senescence-like phenotype, and chronic neurodegeneration in tauopathies, including Alzheimer’s disease. As terminally differentiated cells, neurons may seem incapable of initiating a senescence stress response. However, our data indicate that neurons with mature neurofibrillary tangles are arrested in a cellular senescence-like state. The objective of this project is to identify the upstream molecular mediators and downstream cellular consequences of neuronal senescence in the human brain. High resolution profiling methods will be applied to analyze neurons across the adult human lifespan, and throughout the progressive stages of Alzheimer’s disease. This project will significantly advance the basic understanding of this novel neuronal cell fate, cellular senescence, and its influence on brain health. Moreover, the cellular and molecular pathways identified in our project may reveal novel therapeutic targets for intervention, and the age/stage of disease where they would be most beneficial.

NIH Research Projects · FY 2025 · 2020-09

ABSTRACT Fracture nonunion poses a significant clinical problem. In the United States, approximately 1.6 million bone fractures encounter prolonged healing or nonunion each year. Fracture nonunion treatment usually involves complicated and massive procedures in practice, and sometimes needs multiple surgeries, therefore increases the cost of health care and results in marked patient disability. The major population bearing with these clinical complications are patients with inflammatory conditions, e.g, elder patients, smoking, diabetic or rheumatoid arthritis (RA) patients, highlighting the potential deleterious role of chronic systemic inflammation in fracture r epair. The overarching hypothesis of this proposal is that chronic inflammation results in fracture nonunion through Dnmt3b downregulation mediated angiogenesis defect, and local delivery of OPN and CXCL12 restores angiogenesis and fracture repair under inflammatory conditions. This hypothesis is supported by our preliminary data wherein we show that Dnmt3b is highly expressed in fracture callus during fracture repair and Dnmt3b is the major DNA methyltransferase (Dnmt) responsive to cytokines in MPCs. Relevant to our proposal, we provide evidence that inflammatory signals inhibit Dnmt3b in an NF-κB-dependent manner. Consistently, mice with Dnmt3b loss-of-function (LOF) in chondrocytes display impaired angiogenesis and fracture repair; and Dnmt3b gain-of-function (GOF) in chondrocytes shows protective effect from inflammation in vitro and accelerates fracture repair in mice. Mechanistically, angiogenesis defect mediated by inflammation and Dnmt3b LOF coincide with downregulation of OPN (Osteopontin) and CXCL12 (C-X-C Motif Chemokine Ligand 12) and exogenous OPN and CXCL12 can restore angiogenesis capacity in vitro. To further examine the efficacy of local delivery of OPN and CXCL12 in vivo, we have developed an optimized biomaterial sheet loaded with OPN and CXCL12 and showed a robust angiogenesis process and a restoration of fracture union in RA mice. Three main Specific Aims are proposed. Specific Aim 1 will delineate the mechanism by which inflammation reduces angiogenesis via downregulating Dnmt3b during fracture repair. Specific Aim 2 will determine the optimal release kinetics of OPN and CXCL12 on angiogenesis. Specific Aim 3 will determine the therapeutic effect of sustained OPN and CXCL12 release on angiogenesis and fracture nonunion in mice. The proposed studies will enhance our understanding of mechanisms by which systemic inflammation (via the NF- κB pathway) affects the angiogenic process through Dnmt3b. This work will establish an important therapeutic option to improve the angiogenesis and fracture healing.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY/ABSTRACT Direct electrical stimulation (DES) of the basolateral complex of the amygdala (BLA) can improve declarative memory, reflecting the role of the BLA in modulating memory processes in medial temporal lobe (MTL) regions as a function of emotional arousal. Thus, DES can reveal mechanisms of BLA-mediated memory enhancement relevant to human mental health and disease. DES of the BLA can be used to interrogate the function of memory circuits, especially how neuronal oscillations in the MTL support declarative memory. First, BLA is hypothesized to wield the capacity to prioritize long-term retention of information initially encountered adjacent in time. Second, the BLA preferentially projects to anterior MTL regions and thus is hypothesized to preferentially modulate memory processes in those anatomic regions, processes thought to support memory for non-spatial items more so than memory for spatial locations. Third, although emotional arousal, amygdala activity, MTL activity, and memory performance are typically correlated, we hypothesize that DES will reveal that BLA outputs to other MTL regions cause improved memory performance by directly eliciting pro-memory oscillatory states in those networks. The expected outcomes represent a significant advancement for the basic science of normal memory function and significant movement towards novel therapeutics designed to emulate endogenous mechanisms of memory enhancement.

NIH Research Projects · FY 2024 · 2020-09

PROJECT ABSTRACT Dr. Cecilia Pascual-Garrido, MD, PhD is a hip preservation and adult reconstruction surgeon whose research goal is to make significant and meaningful contributions in the care of young adults with pre-OA hip disorders, through the process of basic scientific discovery and its translation to patient care. Her educational, clinical and research experiences uniquely prepare her to pursue this work, and this career development award will facilitate the complementary musculoskeletal research training she requires, with an emphasis on epigenetics, transcriptome, bioinformatics and statistical concepts. All activities will occur at the Washington University School of Medicine, an institution with strong health services research program and musculoskeletal research group. Recent reports indicate an etiologic role of Femoroacetabular Impingement (FAI) in up to 50% of OA cases. Over 300,000 individuals in the US undergo primary total hip replacement (THR) annually, with THR utilization projected to double by 2030. While characterization of hip bone deformity in FAI has been extensively studied, the role of intraarticular inflammation and epigenetic changes plays in this disease is unknown. To address this need, the purpose of this project is to identify critical biologic events that are mediators of the OA cascade in hip FAI. Aim 1 will identify a catabolic state in articular chondrocytes (ACs) from the impingement zone in hip FAI. ACs from the impingement zone in FAI (early stage) and OA (late stage) will be tested in cultures with TGFβ and IL-1β. We hope to characterize a catabolic state and abnormal DNA methylation in pathological cells. Aim 2 will characterize spectrum of disease (normal-FAI and OA) through gene expression and DNA methylation profile based. Our preliminary data shows enriched biological processes in hip OA compared to FAI. Additionally, we will investigate that genome-wide methylation profiling in hip OA and hip FAI. We believe that integrating epigenomic and transcriptome data will allow us to better understand how the identified loci may contribute to OA pathogenesis. The application of ATAC-seq in OA is novel and could have tremendous clinical implication to identify altered promoters and enhancer genes that might be involved in the pathogenesis of hip OA. Finally, Aim 3 will characterize a small animal model of hip FAI and secondary OA. This animal model could provide a novel opportunity to test future interventional studies for the treatment of hip OA. Through established collaboration with musculoskeletal researchers and clinician scientists in our institution, this application has the potential to uncover early pathological pathways associated with hip OA secondary to hip FAI, which may have important diagnostic and clinical implications.

NIH Research Projects · FY 2024 · 2020-09

Project Summary Research has yet to understand why some with psychotic-like experiences (PLEs; early markers of psychosis risk) transition to psychosis spectrum disorders whereas others report only transient PLEs. This information will be critical for understanding the etiology of psychosis spectrum symptoms and for prevention and intervention efforts for this major public health concern (~90% of individuals with significant PLEs report mental health diagnoses in adulthood). According to the expanded proneness-persistence-impairment (PPI) model, potential distinguishing factors between transient PLEs and those transitioning to psychotic disorders is whether they are sustained and distressing (i.e., sustained dPLEs). Consistent with NIMH Strategic Objective 2, this K23 application will fill critical missing gaps in the literature by characterizing the key risk factors and clinical significance of early sustained dPLEs. The application will focus on ~11,800 children from the Adolescent Brain and Cognitive Development study initially aged 9-11-years-old followed annually over the course of the award. The analyses will test PPI model hypotheses, including investigating the most important factors distinguishing sustained from transient dPLEs, examining neurobiological correlates (e.g., resting state functional connectivity, cortical thickness, cognitive functioning), family history of psychosis, motor and speech developmental delays, and environmental predictors (adverse childhood experiences, cannabis use; Aim 1). Models will also test whether longitudinal changes in cognitive, neural predictors, and environmental risk factors distinguish sustained versus transient dPLEs (Aim 2). Lastly, the application will also fill a critical research gap by examining the clinical significance of sustained dPLEs, examining the social and educational functional impairments, treatment seeking behavior, and conducting additional data collection when the youth are ages 16 to 18 to assess the base rates of attenuated psychosis syndrome (APS) among youth endorsing sustained dPLEs (Aim 3). To assess rates of APS, the applicant and a masters-level clinician will interview a subset of ABCD participants (n=500) and their parents/caregivers using the Structured Interview of Prodromal Syndromes. Overall, the applicant will implement rigorous practices, including running all analyses for the aims and hypotheses outlined below on two-thirds of data and then replicating the exact same models on an untouched one-third of data. Under the mentorship of a diverse team of experts of developmental psychosis spectrum psychopathology, longitudinal analyses, machine learning, and neuroimaging analyses, this scientifically rigorous proposal will test hypotheses regarding cross- sectional and longitudinal predictors of sustained versus transient dPLEs for the future application of early identification and preventative interventions. The application addresses several gaps in the applicant’s training that are critical for success as an independent clinical investigator, including the need for further training in advanced statistical techniques (e.g., machine learning, longitudinal analyses, neuroimaging analyses) and increased exposure in the area of developmental psychosis spectrum psychopathology.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY Infections due to antibiotic resistant organisms (ARO) are a major threat to public health worldwide and a significant cause of healthcare-associated infections (HAI). AROs are especially threatening to immunocompromised patients in intensive care units (ICUs), where a weakened immune system combined with cancer chemotherapy and antibiotic exposures results in increased risk for HAIs due to AROs. A potential source of ARO transmission is the hospital environment. AROs can survive on objects and surfaces in the ICU for prolonged time intervals, resulting in fixed reservoirs for ongoing ARO transmission to vulnerable patients. To create and optimize infection prevention strategies, it is crucial to understand the link between the hospital environment and HAIs, and to optimize environmental hygiene intervention strategies to prevent HAIs due to AROs. To characterize the environmental areas with the greatest ARO burden in the ICU, our group cultured, identified, and created a biospecimen repository of 1,109 potential AROs from collected from ICU surfaces, patient fecal samples, and clinical bloodstream infection (BSI) isolates from patients admitted to the bone marrow transplant (BMT) ICU. Our preliminary selective microbiologic culture techniques and whole genome sequencing (WGS) data indicate that there is a genetic link between AROs isolated from sink drains and clinical BSI isolates. This raises the hypothesis that AROs in sink drains are being transferred between the ICU environment and patients (or vice-versa), and that the environment may be a reservoir for ARO transmission. Based on these preliminary data, our goal is to define the link between AROs found in the ICU environment and patients with HAIs, and to design and test interventions to reduce the burden of AROs in the ICU environment. In this project, we will utilize antimicrobial susceptibility testing and WGS to longitudinally determine the presence and transmission of AROs across the hospital environment, and determine the genetic link between these strains to those causing BSIs in patients in a BMT ICU. Further, we will implement and measure the impact of a bleach and hydrogen peroxide-based environmental hygiene intervention on the concentration of AROs present in ICU sink drains. Completion of these specific aims will allow us to extensively characterize the transmission dynamics of AROs across environmental surfaces and patients, and to evaluate an environmental hygiene intervention that focuses on sink surfaces and drains.

NIH Research Projects · FY 2024 · 2020-09

Summary Acute kidney injury (AKI) has a wide spectrum of outcomes from recovery to a long-term transition to chronic kidney disease (CKD). Between 2000 and 2014, AKI hospitalizations have increased from 3.5 to 11.7 per 1000 persons. Medicare patients aged 66 years and older hospitalized for AKI have a 35% cumulative probability of a recurrent AKI hospitalization within one year and 28% will be diagnosed with CKD in the same time frame. Men have a higher risk of AKI, and of developing progressive CKD, although the mechanisms are poorly understood. In the mouse, males also show a heightened vulnerability to AKI. Recent single cell RNA-seq studies from the McMahon and Kim groups have highlighted marked differences in gene expression between the sexes in proximal tubule segments, the region of the nephron most susceptible to AKI. Preliminary studies in the Humphreys and McMahon laboratories using single nuclear sequencing identified a cell type resulting from failed repair of proximal tubule cells (FR-PTC) following mild to severe AKI with a pro-inflammatory, pro- fibrotic signature. FR-PTCs are hypothesized to drive progressive kidney disease following AKI. This proposal centers on the postulates that an understanding of sex differences in response to AKI, and the application of genetic approaches to target proinflammatory properties of FR-PTCs and to eliminate FR-PTCs following renal repair, will be effective routes to ultimately benefit patient outcomes post AKI. To this end, we have assembled a complementary team, with prior collaborative experience: Humphreys (Washington University), Kim (University of Pennsylvannia) and McMahon (University of Southern California). All team members have participated in the ReBuilding a Kidney Consortium. In Specific Aim 1: we will characterize successful versus failed proximal tubule repair with single nucleus transcriptomics (snRNA-seq) and single nuclear chromatin accessibility studies (scATAC-seq) in male and female mouse models examining key findings in human kidney biopsies. In Specific Aim 2: we will harmonize multimodal datasets generated in Specific Aim1 to facilitate viewing and interrogation of these data by the broad research community. Mining of these data by the group will focus on defining the regulatory logic of repair strategies and outcomes in the male and female kidney. In Specific Aim 3: we will examine the hypothesis that adverse outcomes in the male kidney following AKI are driven by NF-kB pathway components Nfkb1 and TNIK in FR-PTCs, genetically eliminating the action of these genes. We will generate and validate a new transgenic mouse resource for the community, enabling genetic modification and elimination of FR-PTCs. We will determine whether FR-PTC removal has a favorable outcome, as we predict, on progressive kidney disease following AKI.