University Of Alabama At Birmingham

NIH Research Projects · FY 2025 · 2021-09

Neurodevelopmental processes are shaped by dynamic interactions between genes and environments. Maladaptive experiences early in life can alter developmental trajectories, leading to harmful and enduring developmental sequelae. Pre- and postnatal hazards include maternal substance exposure, toxicant exposures in pregnancy and early life, maternal health conditions, parental psychopathology, maltreatment, and excessive stress. To elucidate how various environmental hazards impact child development, it is imperative that a normative template of developmental trajectories over the first 10 years of life be established based on a sufficiently large and demographically heterogeneous sample of the US population. To accomplish this, the Healthy Brain and Child Development (HBCD) Consortium has been formed to deploy a harmonized, optimized, and innovative set of neuroimaging (MRI, EEG) measures complemented by an extensive battery of behavioral, physiological, and psychological tools, and biospecimens to understand neurodevelopmental trajectories in a sample of 7,200 mothers and infants enrolled at 27 sites across the United States (US). The HBCD Study will carry out a common research protocol under direction of the HBCD Consortium Administrative Core (HCAC) and will assemble and distribute a comprehensive and well-curated research dataset to the scientific community at large under the direction of the HBCD Data Coordinating Center (HDCC). The overarching goal of the HBCD Study is to create a comprehensive, harmonized, and high-dimensional dataset that will characterize typical neurodevelopmental trajectories in US children and that will assess how biological and environmental exposures affect those trajectories. A special emphasis will be placed on understanding the impact of pre- and postnatal exposure to opioids, marijuana, alcohol, tobacco and/or other substances. To address these broad objectives, the sample of women enrolled will include: 1) a varied cohort that is representative of the US population; 2) pregnant woman with use of targeted substances (opioids, marijuana, alcohol, tobacco); and 3) demographically and behaviorally similar women without substance use in pregnancy to enable valid causal inferences. In addition, the HBCD Study will identify key developmental windows during which both harmful and protective environments have the most influence on later neurodevelopmental outcomes. The large, multi-modal, longitudinal, and generalizable dataset that will be produced for the first time by this study will provide novel insights into child development using state-of-the-art methods. The HBCD Study will inform public policy to improve the health and development of children across the nation.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Rates of opioid use during pregnancy are at epidemic proportions. Infants exposed to opioids before birth frequently develop neonatal opioid withdrawal syndrome (NOWS) and are at a much higher risk of sudden infant death syndrome (SIDS). A major knowledge gap is that it not currently possible to determine the magnitude of NOWS or SIDS risk in opioid-exposed infants. Most infants with antenatal opioid exposure are observed for several days after birth in the hospital to determine if the infant develops NOWS. Infants who develop NOWS are frequently managed using the Finnegan scoring system which is complex and subjective. There is therefore a need for rapid objective and quantitative measurement of signs of opioid withdrawal. There is a critical need to determine if the hospital course of NOWS or abnormalities in respiratory control can be predicted soon after birth, in order to initiate earlier monitoring or therapy. Furthermore, it is essential to be able to adjust therapy in a more evidence-based manner. The overall objective of the Vital Signs In Opioid-exposed Neonates (ViSION) project is to use cardiorespiratory dynamics (characteristics and patterns of heart rate, respiratory rate, and oxygen saturations) soon after birth from the existing large cohort of opioid-exposed newborns (>140/yr) at the University of Alabama at Birmingham to develop and validate mathematical models to predict (a) onset of NOWS, (b) changes in magnitude of NOWS as estimated by Finnegan scoring, and (c) abnormal polysomnographic (sleep) study at three months of age. The central hypothesis of this proposed study is that cardiorespiratory dynamics improve detection of NOWS onset and severity, and can predict infants with sleep disordered breathing before discharge. The Specific Aims are: Specific Aim 1: Test the hypothesis that cardiorespiratory dynamics soon after birth identify opioid-exposed neonates at higher risk of neonatal opioid withdrawal syndrome. This hypothesis will be tested in a prospective cohort of 200 opioid-exposed newborn infants and 100 healthy control infants. Specific Aim 2: Test the hypothesis that cardiorespiratory dynamics of infants diagnosed with neonatal opioid withdrawal syndrome track temporally with clinical assessment by the Finnegan scoring. This hypothesis will be tested in a prospective cohort of 100 newborn infants diagnosed with neonatal opioid withdrawal syndrome. Specific Aim 3: Test the hypothesis that intermittent hypoxemia and bradycardia events soon after birth are associated with apnea-hypopnea index on polysomnographic studies at three months of age. This hypothesis will be tested in a prospective cohort of 60 newborn infants diagnosed with NOWS, 60 opioid-exposed infants without NOWS, and 60 healthy control infants.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT The “Deep South,” including Alabama, Mississippi, and Louisiana, has the highest rates of obesity, diabetes and hypertension in the nation. As a result, life expectancy in the Deep South is substantially lower than other regions. The mission of the Deep South Center to Reduce Disparities in Chronic Diseases is to develop a regional research center focused on the prevention, treatment and management of cardiometabolic diseases among populations who suffer disproportionately from these conditions in Alabama, Mississippi, and Louisiana. The Center is unified thematically through the application of the precision public health approach across the care continuum to reduce the burden of cardiometabolic disease, as improving health outcomes will require precision public health, i.e., “providing the right intervention to the right population at the right time.” This approach acknowledges the importance of context, individual beliefs and preferences as well as the need for multi-level and multi-domain interventions. The Center brings together an interdisciplinary team of investigators from 4 institutions in 3 contiguous states (the University of Alabama at Birmingham, Tuskegee University, Louisiana’s Pennington Biomedical Research Center, and the University of Mississippi Medical Center) as well as regional non-academic partners to extend cardiometabolic research into real-world community and clinical settings. The Center will drive academic and nonacademic partners toward a new level of intellectual synergy, collaboration, and sustainable efforts to disseminate effective interventions that improve health outcomes in the region. To achieve the long-term goal of improving population health, the Center will provide and coordinate resources not currently available through the following: 1) an Investigator Development Core, to expand the region’s research workforce through enrichment activities and a robust pilot program; 2) a Community Engagement Core, to promote effective bi-directional collaborations between researchers and non-academic partners; and 3) the Administrative Core, to provide leadership and support for all Center initiatives. The Center also includes three interrelated research projects evaluating multi-level and multi-domain interventions that are informed by, and conducted with, academic and community partners in the region to improve cardiometabolic health outcomes among populations with persistent chronic disease burden. Given the significant burden of cardiometabolic diseases evident in the Deep South, the strong research base present at the partnering institutions, and the potential to expand and focus these energies on health outcomes research, the Deep South Center to Reduce Disparities in Chronic Diseases is ideally situated to inform research, clinical care, and policy to improve cardiometabolic health outcomes in a region of tremendous need.

NIH Research Projects · FY 2024 · 2021-09

The overarching goal of the University of Alabama at Birmingham (UAB)/Tuskegee University (TU) Faculty Institutional Recruitment for Sustainable Transformation (FIRST) Partnership is to create systemic and sustainable culture change to further support inclusive excellence in research at both institutions. Within our theme of Health Disparities, we will build a community of scientists committed to inclusive excellence by recruiting 12 early-career faculty committed to promoting diversity and inclusion and interested in addressing health disparities. These faculty will be hired across 3-4 clusters representing areas of research strength and opportunity across our institutions: cancer, obesity and diabetes, cardiovascular disease, and neuroscience. Program hires will be designated as Benjamin-Carver Scientists in honor of two barrier-breaking investigator leaders – Regina Benjamin, MD and George Washington Carver. Our Scientists will be surrounded by a comprehensive support infrastructure including sponsors, mentors, career coaches, institutional research navigators, and professional development opportunities to help them mitigate the difficulties experienced by new hires and accelerate the development of collaborative networks and peer support. We have targeted our activities and support mechanisms to ensure that they impact all levels of influence in the Social Ecological Model including Individual, Interpersonal, Organizational, Community and Policy. All levels are reflected in the following specific aims: 1) recruit and hire clusters of Benjamin-Carver Scientists who are committed to advancing diversity, equity, and inclusion in areas of shared research need and support them by implementing individual career development plans, enhanced with scientific mentoring, career coaching, and advocate sponsorship; 2) provide enhanced support for new faculty through an Office of Integrated Research Support in which Research Navigators will help new faculty coordinate sponsor, mentor, and career coaching support; connect new faculty to existing support resources; and assist new faculty with logistic, fiscal, and regulatory matters related to proposal development and grants management; 3) provide extended opportunities for professional development and networking, including a diverse and inclusive Research Community Collaboratory to help faculty build a collaborative network of peers; 4) expand a series of diversity and inclusion programs at both UAB and Tuskegee to help foster systemic and sustainable cultural change; 5) institute changes to institutional policies and environment to support sustainable institutional culture change, including initiatives to support diverse and inclusive hiring and retention; and 6) evaluate the UAB/TU FIRST Partnership and work with the FIRST Coordination and Evaluation Center (CEC) to determine whether this cohort and cluster design model of faculty hiring, sponsorship, continual mentoring, and professional development, embedded within programs to create academic cultures of inclusive excellence produces significant change in metrics of institutional culture.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY The overarching goals of this proposal are to define how intrinsic IL-2 production and extrinsic IL-2 signals act in conjunction with helper CD4 T cell subsets to configure the exhausted CD8 T cell pool during chronic viral infections and ascertain how these factors contribute to the restoration of responses following checkpoint blockade therapies. This is significant for providing both fundamental insights into the regulation of exhausted T cell ontogeny as well as for devising strategies to structure this ensemble to improve infection control while avoiding immunopathology. The premise is founded in part on a series of exciting published and preliminary findings showing that the pace of viral control can be predicted by the number of IL-2 producing CD8 T cells present at the peak of the effector phase of the anti-viral response. Moreover, the initial formation of IL-2 producing CD8 T cells is accelerated during the earliest stages of chronic viral infections, but this population fails to amplify, suggesting that they may instead serve as progenitors for the development of transitional and terminally exhausted subsets. We also discovered that IL-2-producing effector CD8 T cells generated during acute infections have superior protective powers and are resistant to full terminal exhaustion following adoptive transfer and chronic viral challenge. Additionally, our preliminary findings demonstrate that the cell-autonomous synthesis of IL-2 attenuates the ability to receive STAT5 signals. Collectively, these findings are consistent with a model in which the initial manufacture of IL-2 enables exhausted precursor formation by restricting STAT5- mediated signals, which are known to drive terminal differentiation. Further, we anticipate that the subsequent extinguishment of IL-2 synthesis restores permissiveness to STAT5 signaling, which prompts the further developmental transition of these precursors into more exhausted sub-populations. Our studies are designed to pin-point how intrinsic cytokine production and extrinsic cytokine signals direct the formation and maintenance of exhausted subsets, and provide new insights into the mechanisms that shape the efficacy of the anti-viral T cell pool during chronic infections. We propose the following specific aims: 1. Define the contributions of IL-2-producing CD8 T cells to the formation of exhausted subsets. 2. Define the influence of functionally distinct CD4 helper subsets on the exhausted pool. 3. Determine the temporally distinct roles of IL-2 in structuring the exhausted pool. Our studies take advantage of innovative and technically robust approaches to deconvolute the roles of distinct cytokine producing subsets of anti-viral T cells during chronic infections. They are designed to impact the field by advancing our understanding of how intrinsic cytokine production and extrinsic cytokine and cellular signals integrate to configure the exhausted CD8 T cell pool and contribute to the control of chronic viral infections.

NIH Research Projects · FY 2024 · 2021-09

Abstract African Americans (AAs) have a high prevalence of hypertension and suffer a disproportionately high risk for cardiovascular disease compared to whites. Apparent treatment-resistant hypertension (aTRH) is a severe form of hypertension defined as requiring ≥ 4 antihypertensive medications to achieve blood pressure (BP) control. An estimated 9 million US adults have aTRH. Given the rigorous clinical work-up needed to rule out pseudoresistance and therefore, diagnose true TRH, the population prevalence of TRH is not known. We propose an ancillary study to the Jackson Heart Study (JHS), a large prospective cohort of AA adults (n=5,306) that will deeply phenotype participants with aTRH, ruling out pseudoresistance (i.e., inaccurate clinic BP measurement, presence of a white coat effect, suboptimal antihypertensive therapy, and poor medication adherence) and secondary causes of hypertension (i.e., primary aldosteronism, obstructive sleep apnea, and kidney disease). The JHS provides a highly feasible setting for the proposed study as it includes a large population of AAs with a high prevalence of hypertension and extensive data collection as part of an upcoming fourth parent study visit, which will facilitate the screening and recruitment of participants for our ancillary study. Based on data collected during the fourth JHS exam, we will identify 400 participants with aTRH and 200 treatment-responsive controls. These participants will undergo a clinical evaluation for TRH including assessment of existing ambulatory BP monitoring (ABPM) data to detect white coat effect and urinalysis of antihypertensive drugs and metabolites to assess medication adherence. Also, participants with aTRH will be compared to treatment responsive controls with respect to secondary causes of hypertension and lifestyle factors. Clinical translation of the study findings will be facilitated by an external panel of experts who co-authored the American Heart Association 2018 Scientific Statement on Resistant Hypertension. Overall, the diagnosis and treatment of TRH is limited by our understanding of pseudoresistance and other factors underlying aTRH. The proposed study is urgently needed to more efficiently diagnose and better treat this severe form of hypertension among African Americans.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Many infants exposed to opioids antenatally develop Neonatal Opioid Withdrawal Syndrome (NOWS). The overarching goal of the HEAL Initiative: Neonatal Opioid Withdrawal Syndrome Pharmacological Treatments Comparative Effectiveness Trial – Clinical Sites (UG1 Clinical Trial Required) as stated in RFA-HD-21-031 is to design and implement a multi-center, comparative effectiveness, randomized controlled trial (RCT) to assess the optimal pharmacological treatment for NOWS. Our clinical site at the University of Alabama at Birmingham (UAB) is a Regional Perinatal Center with a Comprehensive Addiction in Pregnancy Program (CAPP) that can enroll a large cohort of infants with NOWS. UAB is also actively enrolling in the UAB Outcomes of Babies with Opioid Exposure (OBOE) in response to HEAL Initiative: Antenatal Opioid Exposure Longitudinal Study Consortium (RFA- HD-19-025) and the ACT NOW Weaning study. As a center in the NICHD Neonatal Research Network (NRN) for >25 years, our clinical site has long-standing established productive collaborations with many other clinical centers. Our center has an excellent track record of enrollment in clinical studies (#1 or 2 in most NRN trials) with successful follow-up to 2 years of age and beyond, with trained certified examiners for neurodevelopment. Our investigational pharmacy has worked with us on multiple trials. Therefore, there is demonstrated adequacy of clinical, administrative and data organizational management facilities. For this delayed onset clinical trial, the final protocol will be developed in coordination with the other Clinical Sites and the DCC. We clearly express our intent to participate in a cooperative manner with the other Clinical Centers, the DCC, the NIH, and the DSMC in all aspects of research in a manner consistent with the terms of the award. The Specific Aims are: Specific Aim 1: To develop a protocol in coordination with the other Clinical Sites and the DCC, to compare morphine, methadone, and buprenorphine, while taking into account non-pharmacologic factors and adjunct therapies for the management of neonatal opioid withdrawal syndrome. Specific Aim 2: To recruit and enroll infants with neonatal opioid withdrawal syndrome at our clinical site in the randomized controlled trial, and measure important short-term outcomes by hospital discharge. Specific Aim 3: (A) To determine important neurodevelopmental and other clinical, social, and environmental outcomes in enrolled infants at two-year follow-up outcomes; and (B) In collaboration with the other Clinical Sites and the DCC, to disseminate findings of the clinical trial and help develop evidence-based clinical practice guidelines.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY SARS-CoV-2 has infected over 138 million people and resulted in over 2.8 million deaths so far, with the expectation the pandemic will continue for many more months, and the virus will persist endemically for years, exacerbated by emerging variants of concern (VoC). Although several vaccines are being used wide-spread, it is unclear if they will be able to induce effective long-term immunity against emerging VoC. Highly effective anti- viral therapeutics for SARS-CoV-2 remain elusive, although several monoclonal antibodies (mAbs) targeting the Receptor Binding Domain (RBD) of the Spike (S) protein have been granted EUA for mild to moderate infection, their effectiveness against severe disease has not yet been evident. With the slow pace of global vaccination, limited anti-viral use/efficacy, and the emergence of antigenic drift variants, the trajectory of this pandemic and future resurgences of the virus is of great concern. Fundamental understanding of the mechanisms of inducing and sustaining protective humoral immunity to SARS-CoV-2 will be critical to its mitigation. The virus is now classified into several clades, numerous VoC emerging, and indications including our work and others that some of this antigenic drift is the result of the virus escaping from immune pressure and increased transmissibility. Drift within the RBD is of the utmost concern as it can enhance the infectivity of the virus and negate the activity of NAbs that may have developed from previous vaccination or infection. Numerous reports have emerged of repeated SARS-CoV-2 infections in patients, and breakthrough infections in fully vaccinated individuals, highlighting the imperfection of naturally acquired SARS-CoV-2 immunity. Utilizing our rationally designed RBD/RBD-ACE2 fusion protein variants, we have identified epitopic and phenotypic heterogeneity amongst RBD-specific human B cells and have isolated several potent RBD-specific human neutralizing monoclonal Abs (NmAbs) (IC50<50 ng/ml) against SARS-CoV-2 which are entering into a Phase 1/2 clinical trial using inhaled delivery in the coming months. We hypothesize that within RBD, the highly conserved regions (RBD-CR), epitopes desirable for mediating broad and potent humoral protection, are surrounded by variable regions (RBD- VR) that are structurally dynamic and highly susceptible to antigenic drift. Further, we hypothesize that RBD-VR mitigate the development of potent and broad RBD-CR specific humoral responses through their immunodominance and direct occlusion of RBD-CR. This RBD-CR/RBD-VR evolutionary dynamic is likely to regulate the sustained protection (or failure) of humoral responses against future viral variants. We will 1) define the ontological and phenotypic diversity of the human RBD-specific neutralizing antibody response, 2) define the dynamics of maintenance of ACE2 binding and immunological pressure on constraining RBD evolution, and 3) determine RBD Ab tolerance for and contribution to SARS-CoV-2 drift. Defining the limits of natural infection and vaccination induced RBD neutralizing antibodies to drive antigenic drift and confer protection from divergent SARS-CoV-2 viruses will inform the development next generation SARS-CoV-2 vaccines and therapeutics.

NIH Research Projects · FY 2025 · 2021-09

It has been shown that although women tend to smoke less than men, they are less likely to quit tobacco use. We developed and established the efficacy of a theory-based Community Health Worker (CHW) intervention for low-income women that augments the tobacco cessation program offered through the public health system. Through previous NIH funding (R01), we developed and established the efficacy of a theory-based Community Health Worker (CHW)-led intervention for low-income women that augments the tobacco cessation program offered through the public health system. Participants in the intervention had 1.88 times the odds of tobacco cessation than participants in the control condition, which consisted of a scheduled appointment to attend the tobacco cessation program through the public health system (20% vs. 11% cessation, respectively). Although our tobacco cessation intervention was successful, CHWs’ feedback indicated that the program was very time consuming. Thus, we propose an adaptation of this CHW-delivered tobacco cessation program to be integrated with mHealth support through mobile devices (App) for participants and a tracking system for CHWs. The relevance of this approach is that it is theory-based (Social Cognitive Theory) and will be adapted based on an efficacious intervention as well as salient features of other mHealth applications that have been shown to be successful in engaging users. During the R21 phase, we will make the adaptations and determine feasibility of a CHW-delivered intervention that is integrated with a mHealth tobacco cessation application (interactive App for participants and tracking system for CHWs) through formative assessments among all involved, pretesting, and pilot testing of the intervention. In the R33 phase, we will assess the effectiveness of the integrated CHW-mHealth tobacco cessation intervention through a group randomized trial with towns as the unit of randomization (8 towns, N=344). The comparison group will be the same as the previous intervention (consisting of a home visit by a CHW during which the participant is scheduled to attend the tobacco cessation program at the neighborhood public health clinic) to allow for comparisons. We hypothesize that at 6-months, women smokers who receive the integrated CHW-mHealth intervention will have significantly higher 7-day point prevalence abstinence (defined as no cigarettes in the past 7 days) than women smokers in the control condition. Self-report will be verified through measurement of exhaled carbon monoxide levels among 30% of participants. If shown to be effective, this approach could be utilized as a model for a population-based intervention in low-resource settings, including rural and disadvantaged women in the U.S.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Converging evidence indicates that neuronal and network hyperexcitability is an important early event in Alzheimer’s disease (AD) patients. The cellular and molecular basis of this hyperexcitability is a critical area of investigation and the presence of similar hyperexcitability in animal models enables studies to dissect underlying mechanisms. A key insight is that hyperexcitability in both AD patients and mouse models has a strong diurnal rhythm. Emerging data also indicate that neural excitability in the forebrain is normally under control of the circadian clock, which regulates seizure thresholds and susceptibility to epileptiform activity. Circadian variation in cellular function is driven by transcriptional molecular clocks expressed in most cells, and molecular clock ablation increases AD pathology. We have compelling preliminary evidence for rhythmic variation in neuronal excitability that is at least partly due to circadian regulation of the membrane properties of inhibitory interneurons, especially fast-spiking cells expressing parvalbumin (PV). Given that PV+ interneurons in the cortex and dentate gyrus are strongly implicated in AD, and that circadian rhythms are disrupted in AD patients and AD mouse models, we propose rigorous experiments to test the hypothesis that dysregulation of the molecular clock and resulting changes in PV+ interneuron gene expression and activity contribute to AD- related neuronal hyperexcitability. Specifically, we will evaluate the differences in circadian clock and clock- controlled gene expression in PV+ interneurons vs. excitatory neurons in the mouse models of AD, using a combination of RNA sequencing, state-of-the-art bioinformatics, and recently developed tools to evaluate molecular clock rhythmicity and transcription in a cell-specific manner (Aim 1). We will record from inhibitory and excitatory neurons in the dentate gyrus and cortex to determine if clock-driven changes in PV+ inhibitory neuron activity are disrupted in AD models and contribute to overall hyperexcitability (Aim 2). Finally, we will utilize an innovative chemogenetic chronotherapeutic approach to manipulate PV+ interneuron physiology to determine whether reinstating the normal circadian patterns of PV+ interneuron activity in AD mice protects against hyperexcitability, cognitive impairment, and pathology (Aim 3). The proposed studies led by a strong interdisciplinary team use powerful approaches to determine how disruption of circadian rhythms facilitates neuronal hyperexcitability that contributes to early stages of AD. Understanding these mechanisms may catalyze development of behavioral or pharmacologic interventions.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Single amino acid variants in triggering receptor expressed on myeloid cells 2 (TREM2) have been identified by genome-wide association studies to be one of the strongest genetic risk factors for late-onset Alzheimer’s disease (AD). AD-associated variants in TREM2 impair TREM2’s ability to bind and signal in response to endogenous ligands. The molecular mechanisms of TREM2 in AD remain not fully understood, in part due to the lack of conclusive identities for TREM2’s endogenous ligands and the lack of a good understanding of how different endogenous ligands interact with TREM2 to affect TREM2 functions. The central hypothesis is that AD-associated variants at TREM2 basic site allosterically regulate endogenous ligands bound at the hydrophobic site of TREM2, and allosteric co-stimulation by endogenous ligands at both basic and hydrophobic binding sites could enhance TREM2 immune activity in AD. The objective is to identify endogenous ligands that bind at the basic and/or hydrophobic sites of TREM2, determine the allosteric effects of ligand binding at either binding site, and determine the effect of AD-associated TREM2 variants on endogenous ligands binding to TREM2. We will test the hypothesis and implement the objective with three Aims. Aim 1 is to identify endogenous ligands of the basic and hydrophobic binding sites on TREM2. Aim 2 is to determine the allosteric effects of endogenous ligand binding at both TREM2 binding sites. Aim 3 is to determine the effect of AD-associated TREM2 variants on endogenous ligands binding to TREM2. New knowledge could have high impact for AD research and treatment by opening an unrecognized avenue of targeting both TREM2 basic and hydrophobic sites to identify TREM2 endogenous ligands and to promote TREM2-regulated immune activation. Research outcomes could aid the development of novel therapeutic strategies, such as combined drug therapy, to enhance TREM2-mediated cellular activities and to compensate the loss of TREM2 function by AD-associated variants for AD treatment. Although immediate potential application for this study is AD pathogenesis, impact on other neurodegenerative and inflammatory diseases affected by TREM2 activation may be enormous. Innovations include 1) this will be the first study to identify TREM2 endogenous ligands by targeting both TREM2 basic and hydrophobic binding sites in a more conclusive manner using the combined in silico virtual screening, biophysical and biological experiments; 2) Results will unveil novel findings that AD- associated variants at TREM2 basic site allosterically regulate endogenous ligand bound at TREM2 hydrophobic site, and allosteric co-stimulation by endogenous ligands at both basic and hydrophobic sites could enhance TREM2-regulated immune activity in AD, providing novel insight into etiology of AD.

NIH Research Projects · FY 2025 · 2021-09

Project Summary New therapies are urgently needed for patients with malignant gliomas, which are highly invasive and lethal brain tumors. Patients with glioblastoma (GBM) die within 1-2 years of diagnosis despite current conventional therapies, including surgery, radiation and chemotherapy. A major driver of tumor recurrence is the infiltrative nature of the glioma cells into adjacent normal brain, which is driven by activation of a mesenchymal transcription program. This mesenchymal transition is activated through extracellular stimulation of cell surface receptors by growth factors such as TGFβ1. The overall purpose of the present project is to investigate the role of adhesion G protein-coupled receptor B1 (ADGRB1/BAI1) in the mesenchymal switch and invasion, and explore new therapies for GBM based on the related mechanisms. ADGRB1 is an orphan adhesion GPCR specifically expressed in the brain. We previously showed that ADGRB1 expression is significantly reduced in patients with GBM through epigenetic silencing, suggesting that ADGRB1 loss may facilitate tumor formation. Our new preliminary data show that low ADGRB1 expression correlates with invasion and poor outcome in glioma patients. Restoration of ADGRB1 expression in GBM cells suppresses the mesenchymal phenotype in culture and mice xenografts. Our pilot studies further suggest ADGRB1 can inhibit TGFβ1-driven mesenchymal transition through a WxLWxLW motif in its first thrombospondin type 1 repeat (TSR1). This motif mediates ADGRB1 binding to the latent TGFβ1 complex and prevents TGFβ1 maturation. Based on these results, we hypothesize that ADGRB1 acts as a brain tumor suppressor by blocking the TGFβ1-mediated mesenchymal switch and that restoration of its expression with epigenetic therapy will represent a novel therapeutic intervention for GBM. To test our hypothesis, we propose the following aims: (i) define how ADGRB1 negatively regulates the mesenchymal transition and glioma invasion, (ii), determine how BAI1 antagonizes TGFβ1 pro-mesenchymal signaling and (iii) evaluate whether epigenetic restoration of BAI1 expression can inhibit glioma cell invasion in vitro and in vivo, and augment survival post-operation. These studies are important as we identified a specific region in the extracellular domain of BAI1 that antagonizes TGFβ1 maturation and the glioma mesenchymal switch, providing a new mechanism for antagonizing this oncogenic pathway that can be exploited therapeutically. These findings support targeting this new pathway in patients whose cancers are driven by mesenchymal transition.

NIH Research Projects · FY 2025 · 2021-09

Zambia Alabama HIV Alcohol Comorbidities Program (ZAMBAMA) The overall aims of the Zambia Alabama HIV Alcohol Comorbidities Program (ZAMBAMA) are to (Aim 1) test the effectiveness of a transdiagnostic model, Common Elements Treatment Approach (CETA), to reduce unhealthy alcohol use and improve HIV clinical outcomes in rural and not yet reached HIV clinics, (Aim 2) evaluate the mechanisms through which CETA impacts HIV outcomes, (Aim 3) investigate whether the treatment effect of CETA varies by clinical (e.g., presence of comorbidities), demographic (e.g., sex) and/or contextual factors (e.g., Zambia, Alabama), and (Aim 4) examine implementation factors, including cost, related to integrated delivery of alcohol reduction interventions to people with HIV and unhealthy alcohol use at front-line HIV clinics. The P01’s central theme is that, among people with HIV and unhealthy alcohol use, integrated screening and treatment of common behavioral and mental health comorbidities will lower unhealthy alcohol use and improve HIV treatment outcomes. ZAMBAMA will be implemented by a collaborative team that brings together groups of established and early-career investigators working to address scientific gaps at the nexus of HIV, substance use, and mental illness in priority geographical areas in the global HIV response: the region south of the Sahara Desert and the Southern U.S. Together, the team will implement two randomized controlled clinical trials in adults with HIV and unhealthy alcohol use to evaluate CETA’s effects on alcohol use, the HIV care continuum (antiretroviral therapy adherence, engagement and retention in HIV care, viral suppression), and common mental health and substance use comorbidities. Project 1 (CETA HIV Alcohol Reduction Trial in Zambia - CHARTZ) will be implemented at public sector primary HIV clinics in Zambia where existing HIV ‘peer educators’ (i.e., lay counselors) will be trained to provide an alcohol brief intervention (BI) and CETA to participants at in-person sessions. Project 2 (Telemedicine for Unhealthy Alcohol Use in Persons Living with HIV using CETA - TALC) will enroll participants receiving HIV care at community Ryan White HIV/AIDS program- funded clinics across Alabama that are serving not yet reached and rural populations. Graduate-level providers (supervised by a licensed clinical psychologist) will provide the interventions (BI and CETA) remotely using telemedicine approaches that were rapidly expanded during the COVID-19 pandemic. Both research projects will also evaluate implementation factors, to enhance the impact of clinical effectiveness data and both projects will be supported by 3 resource cores: a CETA core for clinical intervention training and oversight, a Methods and Analysis core to harmonize data elements across projects and create synergy in data analyses, and an Administrative core to manage business and regulatory requirements of the P01, monitor and manage overall progress, and promote bidirectional knowledge and idea exchange between Southern U.S.- and the region south of the Sahara Desert-focused investigators.

NIH Research Projects · FY 2025 · 2021-09

This K08 proposal describes a 5-year research training program that will prepare the candidate for a career as an independent, NIH-funded translational scientist focused on mechanisms driving sepsis-mediated vascular disease. The scientific premise for the aims is that vascular endothelial dysfunction is a key driver of organ injury in pediatric sepsis. Morbidity and mortality remain unacceptably high in pediatric sepsis despite advances in modern medicine, principally due to knowledge gaps in the mechanisms driving vascular disease during sepsis. Endothelial cell-derived angiopoietin-2 (Ang-2) has emerged as a critical promoter of vascular injury and organ impairment in sepsis through its antagonism of the endothelial Tie2 receptor. Plasma Ang-2 levels are significantly elevated in children with sepsis and are associated with measures of organ injury and outcomes. Preliminary data suggest that heparan sulfate cleavage from the surface glycocalyx of flow conditioned primary human lung microvascular endothelial cells promotes Ang-2 expression, implicating a novel paradigm by which Ang-2 is upregulated in sepsis. Preliminary data and prior work also suggest that inactivation of liver kinase B1 (LKB1) and downstream adenosine-monophosphate-activated protein kinase (AMPK) may be integral in this process. Further, the candidate discovered that Ang-2 is bound to the surface of exosomes isolated from plasma of septic children, suggesting that exosomal Ang-2 may significantly contribute to vascular disease and organ injury during sepsis. Together, these data support the novel mechanistic hypothesis that enzymatic heparan sulfate erosion from the endothelial glycocalyx during pediatric sepsis upregulates expression of Ang-2 that, when bound to exosomes, has potent vascular destabilizing effects. To test this hypothesis, in vitro, ex vivo, and clinical studies will be performed in the following two aims. Aim 1: Test the hypothesis that glycocalyx heparan sulfate erosion increases Ang-2 expression from flow conditioned human lung microvascular endothelial cells via attenuated LKB1 activity and downstream AMPK pathway signaling. Aim 2: Test the hypothesis that exosomal Ang-2 (a) is biomarker for organ injury and clinical outcomes in pediatric sepsis and (b) promotes endothelial permeability via Tie2 receptor antagonism. Harnessing the combined expertise of the candidate’s mentoring team and utilizing the wealth of resources available at the candidate’s institution, the candidate will train in advanced translational science techniques germane to the completion of these aims, including microvascular flow modeling, nanoparticle tracking analysis and characterization with flow cytometry, and electric cell-substrate impedance sensing. The proposed research program has defined benchmarks that will facilitate the candidate’s career advancement and will culminate in the submission of an R01 to continue uncovering the mechanisms driving vascular disease in pediatric sepsis that will foster the discovery of novel therapeutics.

NIH Research Projects · FY 2024 · 2021-09

ABSTRACT Many individuals with cognitive impairment experience decrements in everyday function that can signal encroaching dementia. For every year that cognitive decline is delayed there is substantial savings in patient care costs and inestimable savings in quality of life for patients and family members. The prevalence of cognitive impairment in chronic stroke is approximately 50%. A number of techniques have been found to improve various aspects of cognition in the training situation. However, their transfer to everyday situations outside the training setting has been limited as is retention. They are therefore of reduced or narrow practical value. A cognitive ability that is improved but not used to enhance real-world performance has limited significance. Pilot work has been carried out here with a novel intervention designed to fill these crucial gaps. The results are very promising. The purpose of the proposed research is to carry out a randomized controlled trial (RCT) with a large enough sample to rigorously evaluate these preliminary results. The new treatment to be tested combines two repeatedly validated rehabilitation methods: the Transfer Package of CI Movement Therapy and Speed of Cognitive Processing Training. The purpose of doing so is to produce a marked enhancement in the transfer of cognitive improvement from the training setting to instrumental activities of daily living (IADL) in everyday situations. Both halves of the new combination therapy are founded on the investigators' own basic research. In both cases, efficacy is supported by positive results from multiple single-site RCTs and a large multisite RCT. However, the effect of Speed of Processing Training has been found to have limited generalization to cognitive-based IADL in everyday situations. In contrast, one of the main strengths of CI Therapy is that it produces a robust and extensive transfer of treatment effect to everyday life situations by means, in part, of its Transfer Package. A second strength of the Transfer Package of CI Therapy is long-term retention of its effect. In our initial work, the Transfer Package procedures that this lab has used effectively to transfer motor gains from the treatment to everyday setting were adapted to transfer cognitive gains from the treatment to the everyday setting. We propose here an RCT with 2 arms: Speed of Processing Training (SOPT) in the lab combined with the TP (TP+) and SOPT in the lab but without the TP (TP-). Participants will be 120 adults more than 1-year post-stroke with mild to moderate cognitive impairment. Performance on cognitive-based tasks will be measured in daily life and in the lab before and after baseline, after treatment, and 6- and 12-months afterwards. This design will permit rigorous testing of the value of the Transfer Package with an adequate sample size, objective measurement, blinding, and between-subjects control for amount and intensity of training and therapist contact.

NIH Research Projects · FY 2025 · 2021-09

Project Summary / Abstract We look between targets located in the 3D visual environment by making disjunctive saccades that bring the target image onto both foveae. Each gaze shift is followed by a fixation period for visual analysis during which the new vergence level must be maintained. Most studies have focused on the circuitry controlling conjugate saccades, whereas the neural control of disjunctive saccades and vergence eye movements has received less study. Several models suggest that abducens motoneurons send a monocular command carrying information to each eye to control disjunctive saccades. Other models have proposed the existence of saccade-vergence burst neurons (SVBNs) that project to medial rectus motoneurons and are active only during disjunctive saccades. We have identified this novel cell type, which only discharge during disjunctive saccades, in the central mesencephalic reticular formation (cMRF) lateral to the oculomotor nucleus (OMN). Electrical microstimulation in this region of the cMRF elicits disjunctive saccades, whereas inactivation impairs vergence gaze holding. Recent anatomical findings have demonstrated that premotor neurons related to the near response are located in the cMRF, and that they project to the supraoculomotor area (SOA) and to the OMN. We hypothesize that the cMRF, and the SVBNs in particular, play a critical role in the generation of disjunctive saccades. We further hypothesize that projections between the cMRF and SOA form part of a previously undescribed neural circuit that produces vergence integration, allowing vergence angle to be maintained during fixation. Other anatomical and electrophysiological findings demonstrate that the cerebellum, specifically, the caudal fastigial nucleus and the posterior interposed nucleus, play a role in controlling vergence eye movements in both normal and strabismic individuals. We therefore hypothesize that the cerebellar input to the cMRF/SOA complex helps encode or modulate disjunctive saccades. Guided by these overarching hypotheses, we propose Specific Aims to characterize this neural circuitry. 1. To determine the role of SVBNs and the cMRF in the production of disjunctive saccades; 2. To test the hypothesis that the cMRF/SOA complex is the vergence integrator responsible for maintaining the level of convergence; 3. To determine how the cerebellar projections to the cMRF/SOA circuitry are involved in the generation of disjunctive saccades and vergence eye movements. To test our specific hypotheses, we will use established neurophysiological techniques (electrophysiological recordings, antidromic activation, electrical microstimulation and reversible pharmacological modulation). The overall goal of our project is to substantially increase our understanding of the neural circuitry controlling 3D eye movements in primates, and to broadly impact the oculomotor field, leading to new neurophysiological and modeling approaches. These findings will also provide a critical basis for understanding the absence of precise binocular coordination in eye movement dysfunctions such as strabismus.

NIH Research Projects · FY 2024 · 2021-09

Project Summary/Abstract Acute exacerbations of chronic obstructive pulmonary disease (COPD) result in significant morbidity and healthcare costs, especially severe exacerbations that require hospitalization. Traditional bronchodilators and anti- inflammatory medications, as well as programs to monitor and treat symptoms, have a modest effect on reducing hospital admissions. Readmission rates remain unacceptably high, so alternative approaches are needed. In this regard, pulmonary rehabilitation (PR) is remarkably effective, and is associated with an 80% reduction in admission rates. Successful completion of PR is also associated with substantial improvements in quality of life, dyspnea, as well as exercise tolerance. Despite these benefits, recent studies have highlighted very poor referral rates for PR overall and as low as 1.9% post-hospitalization, and non-completion rates as high as 60%. The poor adaptation of PR in the community is due to a combination of barriers in availability and accessibility, as well as attrition. The number of PR centers in the United States is inadequate, and these centers are mostly distributed in urban areas. In addition, multiple socioeconomic and medical barriers hinder access to PR, and contribute to high drop-out rates. New strategies are needed to reduce hospital readmission in COPD, and to increase the delivery of PR to underserved urban as well as rural areas in the community. Given the scarcities in existing resources, conventional models of care delivery are being challenged, and attempts are being made to find alternative, cost-effective ways of delivering healthcare to a larger number of eligible patients. To overcome the socioeconomic and physical barriers to PR, we hypothesize that a video telehealth intervention that will deliver PR to the patient's home, regardless of geographic location, will reduce hospital readmissions in COPD, and reduce respiratory morbidity. To test our hypothesis we propose a prospective randomized controlled phase 3 multicenter clinical trial comparing a real-time video telehealth PR intervention plus standard of care versus standard of care alone, with the following Specific Aims: (1) To determine if a video telehealth PR intervention reduces 30-day all-cause readmissions in patients hospitalized for acute exacerbation of COPD, (2) To evaluate the effects of the video telehealth PR intervention on dyspnea and respiratory quality of life in COPD post hospital discharge, and (3) To evaluate the cost-effectiveness of the telehealth intervention. Accomplishment of the aims of this study will result in a significant reduction in COPD readmission rates, and a paradigm shift in the way PR is delivered to patients with COPD, especially those that reside in remote and rural areas with limited access to pulmonary rehabilitation.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Hypertension (HTN) and chronic kidney disease (CKD) overburden African Americans (AAs). These disparities translate to higher rates of cardiorenal disease endpoints including stroke, coronary heart disease (CHD), end stage renal disease (ESRD), and death. Blood pressure (BP) lowering with antihypertensive treatment reduces the risk of these outcomes, but the effects of treatment may be variable in different race groups. Studies have demonstrated that AAs respond best to calcium channel blockers and diuretics and not as well to to beta- blockers, angiotensin converting enzyme inhibitors, or angiotensin receptor blockers in comparison to their European American (EA) counterparts. The reasons for differences in cardiorenal health and antihypertensive treatment response are multifactorial and thought to include both environmental and inherited factors. Prior genetic and pharmacogenetic association studies of HTN and BP response to antihypertensive agents have been undertaken in AAs, but these studies have been considerably smaller in scope and sample size compared to those of EA populations. Smaller samples sizes of existing genetic datasets have hindered polygenic risk prediction in this population with the potential to create new health disparities. In order to overcome the limitations of previous research and enable efforts in personalized medicine in AAs, we will leverage data from existing cohorts for one of the largest genomic and pharmacogenomic studies of cardiorenal traits to date. Our pharmacogenetic discovery includes >4000 AAs randomized to chlorthalidone and >2500 randomized to lisinopril from the GenHAT study, an ancillary study of the Antihypertensive and Lipid Lowering Treatment to Prevent Heart Attack Trial. We have established an agreement with the International Consortium for Antihypertensives Pharmacogenomics Studies (ICAPS) for validation of our findings. Our genomic discovery is anchored in whole-genome imputed GWAS data from ~12000 REGARDS study AA participants and ~5000 AAs (JHS, Genoa, HyperGEN) with relevant phenotype and genotype data from the NHLBI’s Trans-Omics for Precision Medicine (TOPMed) program. We will replicate our top variant-association findings in additional populations (~11,000 AAs) with relevant data followed by polygenic risk score testing in other cohorts from TOPMed. Using these rich resources we will derive new screening tools for antihypertensive treatment response and cardiorenal diseases. Polygenic risk score applications are increasing in other populations and this research will substantially improve the available data in underrepresented AAs. .

NIH Research Projects · FY 2024 · 2021-09

Project Summary/Abstract Acute exacerbations of chronic obstructive pulmonary disease (COPD) result in significant morbidity and healthcare costs, especially severe exacerbations that require hospitalization. Traditional bronchodilators and anti- inflammatory medications, as well as programs to monitor and treat symptoms, have a modest effect on reducing hospital admissions. Readmission rates remain unacceptably high, so alternative approaches are needed. In this regard, pulmonary rehabilitation (PR) is remarkably effective, and is associated with an 80% reduction in admission rates. Successful completion of PR is also associated with substantial improvements in quality of life, dyspnea, as well as exercise tolerance. Despite these benefits, recent studies have highlighted very poor referral rates for PR overall and as low as 1.9% post-hospitalization, and non-completion rates as high as 60%. The poor adaptation of PR in the community is due to a combination of barriers in availability and accessibility, as well as attrition. The number of PR centers in the United States is inadequate, and these centers are mostly distributed in urban areas. In addition, multiple socioeconomic and medical barriers hinder access to PR, and contribute to high drop-out rates. New strategies are needed to reduce hospital readmission in COPD, and to increase the delivery of PR to underserved urban as well as rural areas in the community. Given the scarcities in existing resources, conventional models of care delivery are being challenged, and attempts are being made to find alternative, cost-effective ways of delivering healthcare to a larger number of eligible patients. To overcome the socioeconomic and physical barriers to PR, we hypothesize that a video telehealth intervention that will deliver PR to the patient's home, regardless of geographic location, will reduce hospital readmissions in COPD, and reduce respiratory morbidity. To test our hypothesis we propose a prospective randomized controlled phase 3 multicenter clinical trial comparing a real-time video telehealth PR intervention plus standard of care versus standard of care alone, with the following Specific Aims: (1) To determine if a video telehealth PR intervention reduces 30-day all-cause readmissions in patients hospitalized for acute exacerbation of COPD, (2) To evaluate the effects of the video telehealth PR intervention on dyspnea and respiratory quality of life in COPD post hospital discharge, and (3) To evaluate the cost-effectiveness of the telehealth intervention. Accomplishment of the aims of this study will result in a significant reduction in COPD readmission rates, and a paradigm shift in the way PR is delivered to patients with COPD, especially those that reside in remote and rural areas with limited access to pulmonary rehabilitation.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Hypertension is the most common comorbidity with HIV affecting over two thirds of women with HIV over forty years of age. Hypertension is a prominent risk factor for cardiovascular disease (CVD) that is modifiable through lifestyle behavior changes such as physical activity. On average, women with HIV engage in low levels of physical activity particularly if they reside in the southeastern United States where CVD risk is substantial. There are gaps in knowledge regarding the integration of lifestyle medicine approaches in HIV primary care. Further, under-representation and low engagement are significant barriers to existing behavioral physical activity interventions for women with HIV. The goal of the proposed Mentored Patient-Oriented Research Career Development Award (K23) is to provide the essential training and professional scaffolding critical for Dr. Crockett's career transition to independent clinical scientist focused on preventing and addressing cardiovascular comorbidities in underserved populations. To achieve this goal Dr. Crockett will pursue formal training in hypertension and cardiovascular health outcomes, advanced mixed methods research, adaptation of behavioral health interventions, and intervention science. These training aims will be accomplished through the wealth of institutional resources at the University of Alabama at Birmingham; close mentorship from experts; formal training through courses and intensive workshops; attendance at local, national, and international meetings; and professional development activities. This training will be applied to a research project that aims to: (1) elucidate the barriers and facilitators to daily physical activity behavior among women with HIV and hypertension using a social cognitive theory framework; (2) systematically adapt a prescriptive physical activity intervention with peer support for women with HIV and hypertension; and (3) conduct a pilot clinical trial of the adapted intervention to evaluate its acceptability, feasibility, and preliminary impact on systolic and diastolic blood pressure. The research aims will be accomplished by conducting a mixed methods assessment of daily physical activity behavior through objective measurement with accelerometers and experience sampling with 30 women with HIV and hypertension; using an evidence-based intervention adaptation framework (the “ADAPT-ITT model”) with input from key stakeholders (i.e., women with HIV, healthcare workers) in focus groups, expert consultations, and integration of the existing literature; and a randomized pilot intervention trial with 50 women with HIV and hypertension. Results of the proposed study will form the foundation for an NIH R01 application to evaluate the adapted intervention in a larger scale randomized controlled trial. The complimentary training and research goals will ensure Dr. Crockett's transition to independent clinical investigator and contribute to reducing the burden of CVD while improving quality of life among women with HIV.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY Cystic fibrosis (CF), a life-limiting autosomal recessive lung disease that affects 1 in 3500 children, is caused by mutations in the Cystic Fibrosis Transmembane conductance Regulator (CFTR) gene. Recently FDA-approved CFTR modulators partially improve lung function for the most prevalent mutation (F508del), but other common genotypes, such as premature termination codon (PTC) nonsense mutations, lack an effective intervention. Even in F508del patients, 25% are low responders to the latest triple combination modulator cocktail. We discovered that a small non-coding microRNA called miR-145 regulates CFTR gene expression and blocks efficacy of CFTR modulators. Our previous data show that complete miR-145 antagonism improves F508del therapy. This project will pursue a more precise strategy that utilizes an antisense oligonucleotide (ASO) to sterically impede only miR-145 binding to CFTR without interrupting other pathways. We hypothesize that ASO-directed miR-145 target site blockade improves CFTR correction. We propose 3 Aims: Specific Aim #1: Enhance F508del CFTR correction selectively through ASO blockade of the miR-145 binding site. Specific Aim #2: Bolster CFTR correction through miR-145 manipulation in low responders. Specific Aim #3: Employ miRNA inhibition to augment readthrough efficacy in PTC mutations. Together, these Aims investigate miR-145 inhibition as a novel strategy to improve next- generation CFTR correction. Aim 1 tests the efficacy and selectivity of ASO target site blockade to enhance CFTR modulator response in F508del CF airway epithelial cells and relevant animal models. Aim 2 uses patient-derived nasal samples from low responders to FDA-approved CFTR modulators to test whether miR-145 inhibition boosts individual benefit. Aim 3 investigates using miR-145 inhibition to increase the CFTR substrate available for PTC readthrough in mutations that currently lack an effective therapy. These experiments will examine a novel, highly selective strategy to advance CFTR-directed therapeutics, broadly applicable across CFTR intervention and patient genotype.

NIH Research Projects · FY 2025 · 2021-09

PROJECT ABSTRACT In family caregiving situations, those with Alzheimer’s disease and related dementia (ADRD) are more likely to experience elder abuse and neglect (EAN) than older adults living alone or not requiring assistance with care. Elder abuse and neglect (EAN) is defined as acts committed by a person in a trusted relationship that cause actual or increased risk of harm to an older adults’ health and wellbeing. EAN consists of different subtypes such as physical and psychological abuse and neglect. About half of family caregivers for persons with dementia self- report doing at least one of these three EAN subtypes. This proposal evaluates an evidence-based tele-coaching intervention, Care-Resistant Behavior Internet Training (CuRB-IT) to enhance coping skills of family caregivers and reduce elder abuse and neglect (EAN). CRB includes actions taken by a person with ADRD to resist or refuse assistance with care, such as refusing to open their mouth for oral care or using physical aggression. Our tele-coaching intervention (CuRB-IT), which increases problem-solving coping skills by teaching practical strategies to manage CRBs, effectively reduces CRB-related caregiver distress. We will test the effectiveness of CuRB-IT in a delayed-intervention randomized clinical trial, within a multi-time series approach. Caregivers in immediate- and delayed-intervention groups (N=266) will complete 4 waves of instrument completion and daily diary surveys describing the frequency of CRB and EAN over a 21-day period at baseline, 3 months, 6 months, and 9 months. Delayed-intervention caregivers will receive weekly texts to reduce inflated intervention effects inherent in many wait-list control designs.22 After the 3-month follow-up, the delayed-intervention group will then receive the CuRB-IT intervention. This design allows us to maximize power to examine efficacy (between-group), mechanism-of-action, and intervention delay (within-group). Thus, we propose the following Specific Aims: 1) Test the efficacy of an online care-resistant behavior coaching intervention in reducing frequency of EAN (a) among the experimental (immediate-intervention group) as compared to the control (delayed-intervention group) (between groups) and (b) within-person from pre- to post- intervention.; 2) Test hypothesized mechanism of action through multi-level structural equation modeling to assess the relationships between CRB-stress appraisal, use of CuRB-IT problem-focused coping strategies and EAN; and 3) Examine intervention decay at 3- and 6-months post intervention to determine performance of intervention and inform scheduling of booster sessions. We will also assess the efficacy of the CuRB-IT intervention in preventing onset of EAN use by family caregivers as an exploratory aim. This proposal directly addresses high-priority research gaps identified by the NIH Office of Disease Prevention and US Preventative Task Force. Next steps in this research program will focus on translation to practice settings such as the UAB Health system’s Caregiver Bereavement Service Line and Adult Protective Services.

NIH Research Projects · FY 2024 · 2021-09

Recent evidence points to αsyn misfolding and cell-to-cell transmission as critical to neurodegeneration in Parkinson’s Disease (PD) and Dementia with Lewy Bodies (DLB). Key steps implicated in αsyn spread include release, uptake, misfolding, and impaired protein degradation, yet the key molecular mechanisms that regulate αsyn spread are poorly understood. Rab proteins are small GTPase proteins that control protein trafficking and degradation and have been implicated in αsyn pathogenesis. Among the Rab proteins, Rab27b is highly expressed at synaptic terminals in neurons in key brain areas affected in PD and DLB. Rab27b regulates synaptic vesicle (SV) exocytosis and recycling. In non-neuronal cells, Rab27b regulates the distal transport of lysosomes. We recently showed elevated Rab27b levels in αsyn models and in human PD and DLB. Furthermore, we observed that Rab27b KD increases αsyn toxicity by disrupting autophagic flux - pointing to a protective role for Rab27b in cells with high intracellular αsyn burden. Surprisingly, the effects of Rab27b were different in the αsyn fibril model, in which neurons were exposed to extracellular αsyn fibrils: Rab27 KO reduced fibrillary αsyn uptake to prevent αsyn inclusions and neuronal loss. Based on these data, we hypothesize that Rab27b plays two critical yet potentially opposing roles in αsyn handling: 1) Rab27b promotes αsyn autophagic-lysosomal degradation, yet 2) Rab27b can facilitate endocytosis of pathologic, extracellular αsyn as part of its role in SV recycling. Early in disease, we propose that Rab27b upregulation is neuroprotective: Rab27b aids intracellular αsyn clearance. Yet, as disease progresses and extracellular αsyn levels increase, Rab27b’s role in SV endocytosis becomes maladaptive, overcoming any protective function in protein degradation, and aids αsyn cell-to-cell transmission. Our proposed studies will examine the impact of Rab27b and its interactors on two critical biological processes implicated in αsyn pathogenesis. In Aim 1, we will test the mechanisms by which Rab27b promotes clearance of intracellular αsyn, and examine its interactions with other proteins involved in autophagic-lysosomal function. In Aim 2, we will elucidate the mechanisms by which Rab27b aids αsyn entry into neurons and determine its interaction with other synaptic trafficking proteins to mediate this effect. In Aim 3, we will test the overall consequences of Rab27b on αsyn pathology over time in two in vivo αsyn models. A greater understanding of the network of proteins that shape αsyn transmission will have significant impact on the development of future therapies for PD and DLB.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY Few therapeutic options exist for patients with age-related macular degeneration (AMD), and no FDA approved drugs are available for the most common subtype, dry AMD. Longitudinal imaging studies have identified the accumulation of drusen near the fovea as the single greatest anatomic risk factor for progression to severe disease, suggesting that loss of lipid homeostasis in the retinal pigmented epithelium (RPE) and choroid is a critical disease mechanism underlying AMD. Moreover, histologic and molecular studies have shown that inflammation of the macular RPE/choroid is an early, unifying feature of AMD. Yet, the molecular and cellular mechanisms responsible for the accumulation of lipid and inflammation in the RPE/choroid remain poorly understood. Using publicly available bulk and single-cell RNA-sequencing data from 150 human donors, we demonstrate that choroidal macrophages—a critical component of the choroidal ecosystem required for maintenance of the choriocapillaris and RPE function—are decreased and dysfunctional in the macular RPE/choroid in AMD. We detail novel transcriptomic observations related to the molecular underpinnings of choroidal macrophages in humans, demonstrating that they are robustly anti-inflammatory and express several key modules involved in lipid clearance. Of note is the expression of triggering receptor expressed on myeloid cells 2 (TREM2), a liporeceptor that mediates lipid homeostasis and immunosuppression in a range of tissues and is lost in several diseases involving accumulation of extracellular material. Using choroidal tissue from mice, we employ a variety of techniques to validate key findings related to choroidal macrophages. We show that choroidal macrophages are important in maintaining lipid homeostasis in the choroid during times of chronic lipid challenge, as these cells expand and contain nearly all of the excess lipid after 9 weeks of high-fat diet. These findings led us to our core hypothesis that TREM2+ choroidal macrophages modulate lipid homeostasis in the RPE/choroid and suppress inflammation and choroidal neovascularization. We propose two Aims using TREM2 knock-out mice to interrogate the role of TREM2 signaling in choroidal macrophages with respect to lipid metabolism and immune and neovascular suppression in the RPE/choroid. We anticipate the findings from this proposal will expand our understanding of the role of choroidal macrophages in the choroidal ecosystem and will identify TREM2 as a novel drug target in AMD. The proposed training plan for the PI, Seth Fortmann, is sponsored by his mentor, Dr. Maria Grant MD. The overall goal of the training plan is to provide the PI with a strong scientific foundation for a successful career as a physician-scientist ophthalmologist. Included in the training plan are experiences that will help the PI: 1) gain competence in a variety of techniques including bioinformatics, flow cytometry, primary cell culture, and ex vivo imaging of human donor eyes; 2) develop hypothesis-driven research; 3) present data in an oral and written format; 4) effectively integrate research with the clinic; 5) collaborate with other scientists; and 6) responsibly conduct research.

NIH Research Projects · FY 2024 · 2021-09

Project Summary Robust and tightly regulated DNA repair is critical to maintain genome stability and prevent disease development. Eukaryotic DNA is packaged into chromatin that has a profound, yet not well understood regulatory influence on DNA repair, replication and genome maintenance. There is a fundamental gap in understanding how DNA repair pathways are regulated and coordinated within structurally diverse chromatin, and across the heterogeneous genomic landscape. The HELLS (Helicase Lymphoid Specific) is a poorly understood chromatin-associated protein, with an emerging new role in genome maintenance. Mouse HELLS is essential for gametogenesis and proper development of the immune system. Mutations in human HELLS cause severe immunodeficiency syndrome ICF (Immunodeficiency Centromeric Instability Facial anomalies). Despite substantial progress in understanding the molecular functions of the mammalian HELLS in DNA methylation and chromatin remodeling, its role in DNA repair and genome maintenance is poorly understood and remains elusive. The unresolved questions remain whether HELLS regulates multiple DNA repair pathways, and whether it has specialized roles in the repair and maintenance of a distinct genomic loci or domains. We have established and validated fungal model, Neurospora crassa to advance the fundamental understanding of HELLS-mediated mechanisms of genome stability. Our studies reveal a new, previously unrecognized link between HELLS proteins and cellular responses to DNA alkylation damage in fungal and human cells. We hypothesize that HELLS protects cells form alkylation-induced toxicity and plays important roles in the repair and stability of the constitutive heterochromatin domains. This hypothesis is founded based on a strong preliminary data in the fungal model Neurospora and in human cells demonstrating that cells deficient in HELLS exhibit sensitivity to DNA alkylation damage and are deficient in the repair of the constitutive heterochromatin. In addition, we discovered that loss of fungal WDR76 protein in HELLS mutant cells leads to the synthetic rescue of the alkylation sensitivity phenotype, implying that WDR76 acts as genetic suppressor of HELLS deficiency. In Aim 1 we will determine the role of fungal and human HELLS remodelers in the repair of alkylation DNA damage. In Aim 2 we will define precise genomic and chromatin contexts that depend on HELLS for genome maintenance. In Aim 3 we will determine the functional relationship between HELLS and WDR76, a WD40 protein implicated in response to DNA alkylation. Successful completion of the proposed research will define HELLS-mediated mechanisms of genome maintenance, and identify additional regulators and pathways cooperating with HELLS in protecting the cells from detrimental consequences of genotoxic stress. These studies will provide important insights into the origin of the disease- causing chromosomal rearrangements and breaks found in many human diseases, including the ICF syndrome.