Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2025 · 2021-07

Project Summary / Abstract The goal of research in the Leibfarth group is to develop platform synthetic methods that enhance the thermomechanical, adhesion, and degradation properties of polymers while also uncovering mechanistic insights that broadly inform synthetic method development. This goal informs our two complementary research areas that seek to 1) leverage chemo- and regioselective C–H functionalization to enhance the properties of commodity polymers and 2) develop stereoselective polymerization methods that uncover emergent polymer properties from simple chemical building blocks. We have identified a compelling opportunity to make progress on a grand challenge in biomedicine – the discovery of degradable biomaterials with concomitant control of thermomechanical properties, degradation profile, and metabolic fate – by leveraging our expertise at the interface of asymmetric catalysis, C–H functionalization, and polymer chemistry. In the five-year period of this MIRA grant, we propose to develop new catalytic approaches to control the stereochemistry of degradable polymers and establish a domain-selective approach to tune the material properties of polymers post-production. These methods will provide access to heretofore unknown semicrystalline biomaterials where critical parameters such as the chemical composition, polymer molar mass, and percent crystallinity can be systematically varied. The novel polymer structures accessed as a result of this work and the comprehensive evaluation of their degradation mechanism will enable the identification of design rules for the development of new classes surface eroding biomaterials. More specifically, we aim to develop new catalytic methods for the stereoselective polymerization of cyclic monomers through complementary kinetic resolution and enantioconvergent synthetic approaches. We envision that the discovery of previously unprecedented stereodefined polymers can be accomplished by developing a comprehensive understanding of structure–reactivity relationships that determine stereoselective addition of monomers to a reactive polymer chain-end. Further elaboration of this stereoselective polymerization philosophy will enable the pursuit of methods for stereoselective radical polymerization in a controlled manner through chiral Lewis acid catalysis, thus providing a strategy to create highly functional and stereodefined polymers from widely-available vinyl monomers. Complementary to the pursuit of stereoselective polymerization, we have identified late-stage functionalization as an underutilized approach to diversify the properties of existing polymers relevant to biomedicine without resorting to de novo synthesis. This philosophy can be used on current materials in medical devices as well as further expand the utility of the new, stereodefined polymers we discover in the course of our research program. By developing methods to conduct domain- selective C–H functionalization reactions, we propose to systematically tune the water transport into the amorphous phase of semicrystalline polymers and, therefore, influence their degradation mechanism.

NIH Research Projects · FY 2025 · 2021-07

Project summary The latent reservoir is the primary barrier to curing HIV, but little is known about the nature of this reservoir or the molecular mechanisms that regulate it. Increasing evidence suggests that the size and nature of this reservoir is impacted by drugs of abuse. Cannabinoid (CB) abuse, in particular, is prevalent amongst people with HIV (PWH), but the impact of CBs on the latent HIV reservoir has not been investigated. CBs are known to have immuno-modulatory and anti-inflammatory activities through activation of the CB2 receptor that is widely expressed in immune cells, including CD4 T cells. Our hypothesis is that CB exposure during HIV infection alters the size, location and transcriptomic phenotype of the latent HIV reservoir through CB2-dependent activation of the AP-1 transcription factor in CD4 T cells. Consistent with this hypothesis we have recently discovered that cannabinoids promote reactivation of HIV from latency in a T cell model. Defining the impact of CBs on the latent HIV reservoir will be critical to designing appropriate approaches to clear the reservoir from PWH who use CBs. To achieve this goal we propose to use cutting edge methods from the fields of single cell multi-omic analysis to characterize the effect of CBs on the latent HIV reservoir, and to test the functional role of CB-activated pathways in HIV latency. In the R61 phase, we will use a primary cell HIV latency model to define the impact of CB exposure on the transcriptome and epigenome of latently infected cells. Additionally, we will use a cutting-edge single cell HIV assay to quantify the size and location of the intact HIV reservoir in a cohort of CB-using PWH compared to non CB-using PWH. For the R33 phase, we will combine the results from these studies with a detailed single cell genomic analysis of identify CB-regulated transcripts in the PBMCs and CD4 T cells from the CB-using PWH cohort. By integrating these datasets we aim to identify CB-regulated pathways that regulate the size and subcellular location of the HIV reservoir. Overall these results will advance our understanding of how CBs interact with the latent HIV reservoir at the molecular level.

NIH Research Projects · FY 2024 · 2021-07

Project Summary/Abstract Disorders of the brain remain an enormous medical challenge. New platforms that allow functional testing in high-fidelity systems that incorporate patient tissue and high-throughput capacity are urgently needed to bridge gap between cell-based assays and whole-animal testing. We propose a multi-institutional effort to identify, develop, and initiate translation of therapeutic neurological agents using the organotypic brain slice culture (OBSC) platform. Our OBSC models leverage existing cellular and extracellular milieu in the live brain slices to allow rapid, functional testing on brain tissue. Our multi-disciplinary team, which spans three CTSA Program hubs (UNC, Duke, and Univ. Florida), has developed multiple models for neurological disorders and shown the effectiveness of the OBSC platform to discover new mechanisms of disease and identify new therapeutic compounds. We have developed technological innovations for OBSC modeling, including elevated-throughput techniques for brain slicing, viral- and biolistic-based transduction of disease-relevant genes, targeted gene knockdown, real-time monitoring using reporter assays, and incorporation of fresh patient tissue. We now seek to leverage the power of the OBSC platform and experience of our team to broaden the utility of the OBSC platform and ultimately improve the care for patients suffering from brain disorders. Our multi-institutional team will bring together and share new disease models, curated panels of therapeutic agents, unique molecular tool kits, and clinical patient tissue. Within the areas of neurodegenerative disease, brain cancers and ischemic disease, we will add disease-relevant capabilities to the platform, screen therapeutic agents, and enable new immune-based approaches, the fastest-growing area of clinical research, with a particular focus on the brain microenvironment. Approaches to accomplish our aims are: (i) We will use OBSC models of neurodegenerative disease to develop methods for tracking of cell signaling using real-time optical imaging, define molecular pathways mediating inflammatory drivers of disease, and identify effective new therapeutic agents. (ii) We will use fresh patient biopsy samples and OBSC models of aggressive brain cancer to characterize he cellular and genomic heterogeneity of brain tumors, the efficacy of anti-cancer immune therapies, and perform drug screens to identify new therapeutic agents. (iii) Lastly, we will utilize OBSC models of ischemic disease to develop methods for pooled molecular screening, investigate the impact of microglia on stroke progression, and identify new compounds that reduce infarct size. Together, our approaches will create an expandable infrastructure built around OBSC technology, accelerate the discovery of new and effective therapeutic strategies, and initiate translation towards ultimate human patient trials to treat multiple disorders of the brain.

NIH Research Projects · FY 2025 · 2021-06

Project Summary/Abstract Acute flaccid myelitis (AFM) is a poliomyelitis-like illness of children that emerged in the US in 2014. Since then, AFM outbreaks occur August-October every other year, concurrent with outbreaks of enterovirus D68 (EV-D68) infection. Increasing evidence now indicates that EV-D68 is a principal cause of AFM. EV-D68, in the same Enterovirus genus as poliovirus, primarily causes respiratory tract infections. However, the mechanisms of EV-D68 neuropathogenesis and the characteristics of the human immune response to EV-D68 are poorly understood. This five-year research career development award will provide training and development of the skills necessary for the PI to establish an independent research program focused on understanding how EV-D68 causes respiratory and neurologic disease and how human antibodies modify this pathogenesis. Currently the PI is an Assistant Professor on the tenure track in Pediatric Infectious Diseases and Microbiology & Immunology at the University of North Carolina at Chapel Hill School of Medicine. His training to date has focused on the isolation and characterization of human monoclonal antibodies (mAbs) from subjects with prior EV-D68 infection. He will supplement this experience with further training in developing in vivo mouse and ex vivo human tissue models of EV-D68 infection with which to study the pathogenesis of this virus and how human antibodies, both monoclonal and polyclonal, modify pathogenesis. The short-term goal of the proposed studies is to test the central hypothesis that human antibodies prevent EV-D68 from causing AFM but do not protect against respiratory disease. The PI will be mentored by national experts in the study of picornaviruses (Drs. Craig Cameron and Stanley Lemon), human airway epithelial cultures (Dr. Raymond Pickles), mouse models of virus infection (Dr. Mark Heise), and human antibodies (Dr. James Crowe, Jr.). The specific aims are to 1) determine how EV-D68-specific human antibodies modify virus-induced respiratory disease and AFM in a mouse model of infection, to test the hypothesis that antibodies efficiently preclude EV- D68 from causing AFM by preventing dissemination to the CNS; and 2) investigate factors that determine the efficacy of antibody at inhibiting EV-D68 infection in differentiated primary human airway epithelial cells at an air-liquid interface, to test the hypothesis that antibodies poorly protect against initial respiratory mucosal infection. Overall, these studies will help determine whether EV-D68 neutralizing antibodies are a mechanistic correlate of protection from AFM by pinpointing the quantity and quality of antibodies capable of restricting EV- D68 infection to the respiratory tract. Industry collaborators are developing protective mAbs made by the PI as human therapeutic agents, which would be the only available targeted therapy for children with EV-D68 infection. Thus, these studies directly translate to importance in human health in addition to contributing to a more fundamental understanding of disease.

NIH Research Projects · FY 2025 · 2021-06

Abstract The large number of disease susceptibility loci identified from genome-wide association studies (GWAS) is enabling polygenic risk scores (PRS) to deliver on their promise to improve health outcomes and to transform the practice of personalized medicine. The limitations of existing PRS for common diseases and related quantitative traits for blood, cardiovascular, and metabolic conditions in the US necessitates larger sample sizes which now can be obtained using recently developed biobank repositories and electronic health records together with larger population-based cohort studies. Thus, it is critical to optimize PRS performance in as many ways as we can, leveraging knowledge from population genetics. Focused strategies to identify high-impact genetic variants could improve the quality of PRS in populations with high-risk alleles. DNA variants with a signature of natural selection often demonstrate such properties, and have been shown to be enriched among top associations for a number of hematological and immune/inflammatory traits that are important biomarkers for key chronic diseases. We propose to focus our PRS studies on hematological and immune/inflammatory traits and their associated chronic diseases and to extend methods for the development of PRS to accommodate various population genetic parameter estimates, high impact genetic variants, and multiple endophenotypes. Thus, our Specific Aims are: 1) Assemble and harmonize data sets needed to accomplish the goals of the project, including hematological traits (red blood cell, white blood cell, platelet), and immune/inflammatory traits (CRP, fibrinogen, D-dimer) from: Jackson Heart Study, Women’s Health Initiative, BioVU, and GeneSTAR. 2) Extend PRS methods to: a) explicitly model population genetic parameters; b) accommodate large-effect risk alleles (such as those from regions with a signature of natural selection); and c) enable joint modeling of multiple endophenotypes; and 3) Develop and apply novel PRS and overall disease prediction models to: a) estimate risk of common diseases and related biomarkers affected by hematological, thrombotic and immune/inflammatory biology; and b) enable calculation of PRS-adjusted clinical laboratory values.

NIH Research Projects · FY 2025 · 2021-06

Project Summary/Abstract Neurons are capable of activating pathways that induce either the degeneration of the entire cell by apoptosis or to selectively degenerate only the axons by pruning. While the main components of the caspase activating machinery during apoptosis and pruning have been identified, a fundamental question of whether the apoptotic machinery is activated throughout the neuronal soma and axons, or is spatially restricted, during these events remains unknown. This question is most relevant in the context of pruning where one predicts the apoptotic machinery to be localized to the targeted axons undergoing degeneration. However, we were surprised to find an unexpected spatial restriction of caspase activation even during apoptosis, where we found these to be restricted primarily to the soma, even though both soma and axons degenerate. In this proposal, we will mechanistically examine the spatial localization of the apoptotic machinery in neurons during apoptosis and pruning. We will utilize neurons cultured in microfluidic chamber devices to allow for the spatial segregation and manipulation of neuronal somas and axons. Our hypothesis is that during apoptosis and pruning, the restricted caspase activity causes the “physiological axotomy” of axons, activating the Sarm1-mediated axotomy pathway of axon degeneration. The concept that the developmental pathways of apoptosis and pruning can cause axotomy is novel because these pathways were considered to be distinct from the injury-induced axotomy pathway. In Aim 1, we will define the spatial restriction of the apoptotic machinery in neurons during apoptosis and axon pruning. In Aim 2, we focus on examining the function of Sarm1 during apoptosis and axon pruning. In Aim 3, we will investigate the Sarm1-deficient mice for pruning defects in vivo and evaluate if these mice exhibit behavioral deficits. This project opens exciting areas of research not only because of its new concepts for apoptosis and pruning, but also because it brings into focus a developmental function of Sarm1 that is beyond its recognized role in axon injury.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY A major barrier in the field of opioid research is our limited understanding of the organization of the opioid system in the brain: we still do not know which cell types express each opioid receptor (mu, delta, kappa, nociceptin) to mediate the effects of endogenous and exogenous opioids, nor what other molecules are present in these cells. Without this knowledge, understanding how opioids alter activity in circuits to produce behavioral effects remains elusive. This gap in knowledge prevents the identification of molecular targets to potentiate opioid analgesia and mitigate the deleterious effects of opioid use disorder (OUD), including addiction and respiratory depression. To fill this gap in knowledge, we propose to leverage the uniquely massive single-cell RNA sequencing (scRNA-seq) database generated by the Allen Institute for Brain Science (>3.4 million cells throughout the entire mouse brain) as part of the BRAIN Initiative Cell Census Network (BICCN) effort. Using this database, we will establish a comprehensive catalog of all the cell types that express each opioid receptor and peptide throughout the brain, as well as the co-expression of gene networks that mediate or regulate opioid actions, including other G protein-coupled receptors and cellular effectors of opioid receptors (Aim 1). Further, we will use our well- established high-throughput single-cell RNA-seq pipelines to characterize the cell-type-specific molecular adaptations that occur during chronic opioid exposure, withdrawal, and abstinence, using a clinically relevant model of post-surgical pain for which opioids are typically prescribed (Aim 2). Finally, we will leverage this novel knowledge to dissect the mechanisms of action of opioids by performing advanced circuit mapping and in vivo functional imaging studies of cell types expressing opioid receptors in the prefrontal cortex (PFC), a region critical to both opioid analgesia and addiction (Aim 3). Our transformative work will be the first to combine several highly innovative technologies at the molecular, circuit, and neural ensemble levels, including high-throughput scRNA-seq, new viral strains with improved transsynaptic transfer and decreased toxicity for circuit mapping, the crystal skull and miniscope- microprism optical approaches for in vivo wide-field imaging of brain state and for recording dynamics of molecularly defined neuron types in freely moving mice undergoing opioid analgesia and addiction paradigms. We have an extraordinary interdisciplinary team of investigators with highly complementary expertise in the neurobiology of opioids and the distribution and function of their receptors in pain and addiction circuits, the molecular and anatomical brain architecture, large-scale cell type characterization and circuit mapping, and highly innovative brain imaging methods as applied to the study of neural circuits. Overall, this research aims to generate an exceptional resource for the opioid field (to be made publicly available through the NIDA-funded SCORCH data coordination center) to explain how opioids change the brain, and discover novel therapeutic approaches to prevent and treat OUD.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY Globally, there are more deaths from cancer than from HIV, tuberculosis, and malaria combined. In 2018, there were more than 18 million new cancer cases and 9.5 million cancer deaths worldwide, and, by 2030, these numbers will increase by at least 50%. Countries with low or medium Human Development Index account for more than 60% of cancer cases and 70% of cancer deaths. Cancer deaths in sub-Saharan Africa occur disproportionately from cancers that are preventable and/or curable including lymphoma, cervical cancer and breast cancer. Addressing the substantial burden of cancer requires a new generation of cancer researchers who can test hypotheses in the African context. The University of North Carolina Project- Malawi (UNCPM) has a rich cancer research environment that includes participation in the National Cancer Institute (NCI) U54 consortium for HIV associated malignancies, the AIDS Malignancy Consortium, the International Epidemiologic Databases to Evaluate AIDS (IeDEA), NCI African Esophageal Cancer Consortium, and a USAID-funded cervical cancer screening study. To date, UNCPM has had significant success in cancer clinical trials and translational research projects, and in establishing a handful of early- career investigators, but momentum is needed to build on this foundation. First, large improvements in cancer control in Malawi require approaches that extend beyond clinical trials to cancer outcomes research, a broader approach that requires unique skills. At this time, there are few cancer researchers with both the necessary cancer research skills and the local experience to develop cancer outcomes solutions adapted to the Malawian context. Second, there is no defined pipeline for early-career cancer research capacity. Finally, there are few opportunities for successful Malawian investigators to obtain the additional skills and experience necessary to be competitive for independent research funding. To address these gaps, we propose to establish the Malawi Cancer Outcomes Research Program (M-CORP). This application is led by UNC in conjunction with the University of Malawi College of Medicine (COM) and in collaboration with other national and international institutions and training programs. M-CORP aims to increase the cadre of trained investigators who can pursue independent research careers by: 1) establishing a robust curriculum and training opportunities across the continuum of cancer outcomes research specific to the Malawian environment and research priorities; 2) supporting advanced degrees; and 3) overcoming obstacles to research independence by providing post- doctoral opportunities including mentored pilot grants. We will use a multi-PD leadership strategy based at both UNC and UNCPM that will optimize our strengths across the spectrum of cancer outcomes research. An exceptional pool of multidisciplinary mentors, a detailed plan for program evaluation, and robust, broad-based partnerships between UNC and Malawian institutions ensure that M-CORP objectives will be met and will establish Malawi as an internationally recognized center of excellence for cancer outcomes research.

NIH Research Projects · FY 2025 · 2021-06

Project Summary The existence of active adult neurogenesis in mammals, including humans, suggests striking structural plasticity and regenerative capacity within the mature nervous system. Adult-born granule cells (GCs) derived from radial neural stem cells (rNSCs) within the dentate gyrus (DG) have been shown to play a critical role in specific forms of memory. Impaired memory, commonly associated with Alzheimer’s disease (AD), correlates with impaired rNSC behavior and hippocampal neurogenesis in AD mouse models and human patients, likely due to lack of permissive niche environment to support neurogenesis. Therefore, identifying critical niche components capable of maintaining NSCs and promoting sustainable neurogenesis will enable development of novel strategies to enhance functional repair from endogenous NSCs. Increasing evidence from human studies have provided tremendous support for the alterations of cholecystokinin (CCK) system in AD patients. Despite these promising findings, the functional role of CCK in heathy and AD brains remains unknown. Our goal is to explore the unprecedented role of endogenous CCK in regulating neurogenic niche and neurogenesis in normal and AD mice. This proposal is built upon a series of our recent findings. Specifically, we found that stimulating DG CCK interneurons to increase CCK level in the DG provides a permissive niche environment to support rNSC proliferation and production of proliferating progeny through the trophic effects of CCK on dentate astrocytes in promoting their glutamatergic gliotransmission. In contrast, reducing dentate CCK disrupts neurogenic niche by inducing reactive astrocytes and neuroinflammation, which correlates with decreased activation of rNSCs and production of proliferating progeny, suggesting an anti-inflammatory role of CCK in DG. Interestingly, 5xFAD mice exhibit dystrophic CCK neurites and reduced dentate proCCK expression, which correlates with reactive astrocytes, impaired neurogenesis, and memory deficits. These data suggest that AD pathology may interact with dentate CCK interneurons to impact various functional aspects associated with DG. These interesting findings sparked the following directions we would like to pursue. Aim 1 is to test the hypothesis that dentate astrocytes mediate CCK- dependent regulation of NSCs and neurogenesis through glutamatergic gliotranmission from astrocytes; Aim 2 is to test the hypothesis that reduced dentate CCK impairs NSC proliferation and neurogenesis through reactive astrocytes mediated interferon-γ signaling onto NSCs; Aim 3 is to test the hypothesis that increasing dentate CCK restores impaired neurogenic niche, neurogenesis, and memory in 5xFAD mice.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY/ ABSTRACT There is a lack of measures available for domains of importance to individuals and families living with dementia. Available measures that do have clinical importance may include abstract constructs and/or complex response formats (e.g. 7-point Likert-type or numerical scales) that are not optimal for persons with dementia. Interdisciplinary researchers are engaging in innovative, person-centered interventions grounded in therapeutic optimism but are constrained by instruments that primarily quantify negative behaviors, deficit and decline, and/or by measuring broad constructs such as global quality of life. The NIH-PROMIS measures demonstrate the value of standardized, common datasets but they do not currently incorporate patient preferences and may not encapsulate the full range of positive outcomes. Rigorous, multi-site psychosocial intervention trials would benefit from a common portfolio of robust measures sensitive to change that capture modifiable and meaningful aspects of living well with dementia over time. This project will develop the infrastructure for development, standardization and validation of new outcome measures and methods for psychosocial interventions in dementia. Emphasis will be placed on measures that are clinically meaningful to persons living with dementia as well as measures that capture modifiable elements of living well with dementia. Measures that optimize longitudinal evaluation of psychosocial intervention for persons with co-occurring cognitive and sensory challenges will be prioritized. In the R21 phase, an interdisciplinary steering council comprised of persons living with dementia, researchers and biostatisticians will convene focused expert panels and collaborate with an advisory board of researchers and clinicians. Using the principles of human-centered design, the council will identify priority outcome measures and create standards for design and testing of novel measures, methods and technologies. In the R33 phase, prioritized measures and methods will be pilot tested. These will include self-report, carer-informed and observational measures. Products of this work will include: 1) Research guidelines for the development and testing of new measures for psychosocial intervention research and 2) Promising new measures, methods and technologies available for larger scale testing with the goal of adding to the NIH- PROMIS measure set. The dissemination of these products will enhance the existing research infrastructure and accelerate progress in psychosocial intervention research.

NIH Research Projects · FY 2025 · 2021-06

SUMMARY My research program focuses on quantitatively characterizing the role of RNA structure in post-transcriptional regulatory processes. Comparisons of protein and messenger RNA (mRNA) abundance at genome scale reveal low correlation between the two gene expression levels in most human tissues and other organisms. This poor correlation suggests that a significant amount of gene regulation occurs post-transcriptionally. To discover elements in mRNAs that control their activities, we measure the effects of human disease-associated structure variants that map to non-coding regions of the transcriptome. Specifically, we integrate computational structure prediction with high-throughput allele-specific chemical structure probing in vivo to assess the functional consequences of RNA structure change. We also establish the causality of these variants by using quantitative reporter assays to measure translation efficiency, splicing, and mRNA stability. In total, these experiments provide molecular explanations of disease mechanisms. To support these goals, we develop, implement, and apply both computational and experimental approaches to study RNA structure in the cell. Our proposed research program will further develop two important technological innovations. The first is a hybrid experimental/computational approach for studying precursor and mature mRNA structure simultaneously in vivo; this approach integrates mutational profiling of SHAPE-MaP data (Selective 2’-Hydroxyl Acylation by Primer Extension – Mutational Profiling) with Boltzmann suboptimal sampling of the secondary structural ensemble. The second innovation is SHAPE-JuMP, which uses a bifunctional RNA modification reagent and a highly processive reverse transcriptase that “jumps” across chemical crosslinks to probe through-space three-dimensional contacts in RNA. We will use these technologies to establish the structures of both precursor and mature mRNAs. In addition, we will extend the biological scope of our work by using these technologies to collaboratively investigate inter- and intramolecular interactions in positive strand RNA viruses. In sum, this program will identify novel RNA structure motifs that regulate the functions of precursor and mature mRNAs and viral genomes.

NIH Research Projects · FY 2024 · 2021-05

Iterative Design to Engage All Learners ABSTRACT In 2016, high levels of per- and polyfluoroalkyl substances (PFAS) were discovered in the Cape Fear River in eastern North Carolina (NC), part of NC's largest watershed and drinking water supply for over 1 million people. More recently, PFAS were detected in central NC, in the Haw River, which supplies drinking water for Pittsboro, NC, and in Jordan Lake, a recreational reservoir that also provides drinking water for several cities in the Research Triangle area. Although PFAS contamination has been detected in 49 states, biomedical researchers in NC are leading the way nationally in assessing the extent of PFAS contamination in waterways and air and in conducting studies on the effects of PFAS on human and ecosystem health. These chemicals have been used since the 1950s in a wide range of consumer products and have been found in the blood of people and animals worldwide. Research suggests that PFAS are harmful to human and animal health, with documented immune system impacts that may influence individual susceptibility to COVID-19. Using cutting- edge, interdisciplinary research on PFAS as a foundation, the Center for Public Engagement with Science in the UNC Institute for the Environment, proposes Iterative Design to Engage All (IDEA) Learners, with a goal of building the capacity of NC teachers, especially those in economically disadvantaged communities and those impacted by PFAS contamination, to introduce current biomedical science and career opportunities to diverse students. We will accomplish this goal through three specific aims: (1) Apply design thinking and design-based research approaches to co-develop, implement, and revise standards-aligned curriculum units that feature current research on the health effects of PFAS; (2) Increase teacher knowledge of current PFAS research and self-efficacy for incorporating current biomedical science into classroom instruction; and (3) Support participating teachers in promoting biomedical research careers to diverse students. Over five years, IDEA Learners will result in two curriculum units with up to eight PFAS-focused lessons that have been designed for NGSS and incorporate research-generated data and science and engineering practices relevant to biomedical research careers. These units will include videos that highlight diverse environmental health sciences researchers. Through long-duration professional development (PD), 48 teachers will deepen their content knowledge and improve their self-efficacy, positioning them to increase URM and female students' interest in current biomedical science instruction. An additional 48 teachers will participate in short-duration PD, and all teachers will have the potential to reach over 30,000 students during the project period. The project will lead to enhanced capacity to offer inclusive learning environments and improved support for URM, female, and LEP students, ultimately cultivating a more diverse future biomedical research workforce.

NIH Research Projects · FY 2025 · 2021-05

ABSTRACT Unintended pregnancy is a major contributor to maternal and infant mortality in low-income countries (LICs). More than 300,000 women and 2.7 million newborns die every year in LICs due to complications from childbirth and pregnancy. Nearly half of the 200 million pregnancies occurring annually in LICs are unintended. High numbers of unintended pregnancy are primarily the result of non-use of contraception. Non-use of contraception is more likely to occur among potential users who experience poor provider care. Providers who are frequently absent, solicit informal payments from clients, and deny methods to unmarried or nulliparous women are a major barrier to women seeking family planning. Yet, removing these barriers is difficult due to low supervision and accountability in under-resourced public facilities. Such findings highlight the need for interventions that increase quality of care via alternative mechanisms for monitoring providers. The social accountability approach solicits citizen feedback with the goal of improving provider performance and service delivery. To date, there is limited rigorous evidence on the effectiveness of social accountability interventions to increase contraceptive use and on the conditions necessary for successful and sustainable scale-up of these interventions. Further, no prior study has rigorously assessed social accountability in a setting where Universal Health Coverage (UHC) is already operating. Based on these knowledge gaps, we propose to evaluate the impact of two social accountability interventions using rigorous methods. We propose this study in Kenya, which rolled out UHC in late 2018 and where one out of every 42 women will die from complications related to pregnancy and childbirth. This study seeks a) to implement the Community Score Card and the Citizen Report Card, b) to evaluate the impact of each of these interventions on contraceptive use, quality of care, and community engagement within communities in Kisumu County, Kenya, and c) to assess the potential for sustainability in additional counties in Kenya, using implementation science methods. To evaluate the impact of the Community Score Card and the Citizen Report Card on our outcomes of interest, a three-armed cluster randomized controlled trial will be conducted in Kisumu, Kenya, with all public-sector facilities randomly assigned to one of three study arms: 1. Community Score Card intervention, 2. Citizen Report Card intervention, or 3. control sites. Outcomes will be assessed via pre- and post-intervention surveys at the individual (n=2268) and facility levels (n=129). Implementation science methods will be used to assess the quality, scalability, and replicability of both the Community Score Card and the Citizen Report Card for uptake by the public-sector healthcare system. Specifically, in-depth interviews will be conducted with community members and service providers (n=30), and focus groups (n=4) will be conducted with key intervention facilitators to assess implementation challenges. This research project will develop an evidence base and implementation strategy for effective community monitoring of publicly funded healthcare facilities in LICs.

NIH Research Projects · FY 2025 · 2021-05

Project Summary Pathological alcohol-seeking behavior is regulated in part by glutamate AMPA receptor (AMPAR) activity in the amygdala. Transmembrane AMPA receptor regulatory proteins (TARPs) profoundly affect the trafficking and function of AMPARs in synaptic and behavioral plasticity. Although the TARP family of proteins is expressed throughout the brain, the TARP γ-8 subtype is restricted to forebrain regions including the basolateral amygdala (BLA); a brain region that is critical to addiction. However, the role of TARP γ-8 in alcohol use disorders (AUD) or other addictions is unknown. To fill this gap in knowledge, we propose an innovative set of behavioral, genetic, bidirectional systemic and site-specific pharmacological, molecular, and physiological studies in mice to evaluate the mechanistic role of TARP γ-8 in alcohol reinforcement, escalated self- administration, and cue-induced reinstatement of alcohol-seeking behavior as a model of relapse. Elucidating the neural mechanisms of these three critical behavioral domains has high translational value for understanding the development, progression, and maintenance of AUD. Successful completion of the studies in this application will provide fundamental mechanistic insights into TARP γ-8 regulation of pathological alcohol-seeking behavior. Moreover, this work moves the field forward in understanding the molecular mechanisms by which alcohol hijacks reward processes and has potential to inform development of new pharmacotherapeutic strategies that target AMPAR function in a highly selective brain region-specific manner.

NIH Research Projects · FY 2025 · 2021-05

Project Summary/Abstract The long-term objective of this research is to improve physical activity among populations experiencing cancer disparities. The goal of this training award is to: (1) deliver a multicomponent, culturally-targeted, technology- delivered, scalable intervention to increase physical activity among African American colorectal cancer survivors; while advancing the development of a new investigator, with a program of research focused on increasing physical activity and decreasing cancer disparities. Specific aims are to: (1) co-create, with AA CRC survivors who engaged in physical activity throughout cancer treatment, culturally targeted narrative videos to increase physical activity knowledge, self-efficacy, outcome expectations, habits and enjoyment; (2) pilot test the intervention to assess reach, effectiveness, adoption, implementation, and maintenance and; and (3) measure outcomes at baseline, 3- and 9-months post baseline. Outcomes include: psychosocial constructs related to physical activity engagement (knowledge, self-efficacy, outcome expectations, enjoyment, habits); physical activity (average daily steps, weekly minutes of moderate-vigorous intensity activity); symptoms (pain, fatigue, depression, bowel dysfunction); and inflammation biomarkers that may explain the pathways through which physical activity impacts cancer outcomes. First, a qualitative exploratory approach will be used to interview 20 African American colorectal cancer survivors. Together with a subset of interested participants a multi-component intervention to increase physical activity among African American colorectal cancer survivors will be created. Next, an additional 72 participants will be recruited to conduct a pilot two-group, randomized repeated measures study to assess Reach, Effectiveness, Adoption, Implementation and Maintenance (RE- AIM) of the intervention. Feasibility of collecting survey and biomarker data at baseline, 3 and 9 months later will be assessed. These timepoints will facilitate exploration of changes pre- and post-intervention, and to determine if effects are maintained 6 months after completing the intervention and chemotherapy. Time since last chemotherapy will be controlled for in all analyses to account for variations in treatment schedules.

NIH Research Projects · FY 2024 · 2021-05

ABSTRACT (Parent Grant) Identification of high-quality chemical probes, molecules with high specificity and selectivity against macromolecules, is of critical interest to drug discovery. Although millions of compounds have been screened against thousands of protein targets, small-molecule probes are currently available for only 4% of the human proteome. Thus, more efficient approaches are required to accelerate the development of novel, target-specific probes. In 2019, a new bold initiative called “Target 2035” was launched with the goal of “creating […] chemical probes, and/or functional antibodies for the entire proteome” by 2035. In support of this ambitious initiative, we propose to develop and test a novel integrative AI-driven methodology for rapid chemical probe discovery against any target protein. Here, we will build an integrative workflow where the unique XChem database of experimental crystallographic information describing the pose and nature of chemical fragments binding to the target protein will be used in several innovative computational approaches to predict the structure of organic molecules with high affinity towards specific targets. The candidate molecules will be experimentally validated and then optimized, using computational algorithms, into lead molecules to seed chemical probe development. The proposed project is structured around three following interrelated keystones: (i) Develop a novel method for ligand-binding hot-spot identification and discovery of novel chemical probe candidates; (ii) Develop novel fragment-based integrative computational approach for accelerated de novo design of chemical probes; (iii) Consensus prediction of target-specific ligands, synthesis, and experimental validation of computational hits. More specifically, we will develop a hybrid method to predict structures of high-affinity ligands for proteins for which XChem fragment screens have been completed. These approaches will be used for screening of ultra- large (>10 billion) chemical libraries to identify putative high affinity ligands within crystallographically determined pockets. Then, we will develop and employ an approach using graph convolutional neural networks for de novo design of a library of strong binders that will be evaluated to select the best candidates for chemical optimization. Finally, we will combine traditional structure-based and novel approaches, developed in this project to select consensus hit compounds against three target proteins: transcription factor brachyury, hydrolase NUDT5, and bromodomain BAZ2B. Iterative design guided by the computational algorithms, synthesis, and testing will progressively optimize molecules to micromolar leads to chemical probes for the target proteins. Completion of the proposed aims will deliver a robust integrative workflow to identify leads for chemical probes against diverse target proteins. We expect that our AI-based computational approach to convert crystallographically-determined chemical fragments into lead compounds coupled with the experimental validation of computational algorithms will accelerate the discovery of new chemical probes, expand the druggable proteome, and support future drug discovery studies

NIH Research Projects · FY 2025 · 2021-05

Project Summary/Abstract The efficient and stereoselective preparation of chiral organic molecules is vitally important to the development of new therapeutic agents. However, despite significant progress, many desirable complex 2D and 3D molecular scaffolds remain difficult to access or are inefficiently prepared. Consequently, development of new transformations that provide synthetic chemists with strategic bond disconnections that facilitate de novo approaches to organic synthesis is of great importance. In addition, such methods should be robust, practical, allow the preparation of useful amounts of material, and ideally employ readily available starting materials. The aim of this MIRA grant is the conversion of an NIGMS funded project with a long-term goal that broadly seeks to develop and exploit new, practical, and efficient stereoselective new technologies for organic synthesis of molecular scaffolds of biological importance that are otherwise difficult to access, or inefficiently prepared via current processes. To this end, we are interested in developing transformations that leverage the broad chemical reactivity of boron-stabilized chemical synthons for complex molecule synthesis. This approach is attractive because reactions of such intermediates permit novel bond constructions and simultaneous installation of a synthetically desirable boron moiety. The proposed studies described in this program will further our expanding studies of catalytic stereoselective C–C bond forming reactions with organoboron reagents. These readily accessible reagents merge powerful bond forming reactivity with broad synthetic utility, to deliver versatile organic compounds of notable significance for fragment-based drug discovery. Building off our previous work, we are developing new catalytic C–C, C–N, and C–X bond forming methods for the synthesis of chiral amines, alcohols, and aromatic structures, with a particular focus on examples that contain quaternary carbon stereogenic centers. We envision that chiral metal nucleophiles can be accessed by enantioselective transmetalation of readily accessible achiral organodiboron reagents. The resultant nucleophilic species can engage in a number of enantioselective reactions with various C=O, C=N, C–X electrophiles. We also aim to target the generation and utility of novel boron-functionalized synthons by metal-free Lewis base catalysis for reactions with electrophiles that are difficult to achieve via transition metal catalysis. Importantly, enantioselective variants of these unknown reactions will herein be explored. Furthermore, new nucleophilic aromatic substitution reactions of aromatic and heteroaromatic compounds with organoboron carbon nucleophiles will be developed that operate under metal-free conditions. Overall, by exploring synthetic reactions of organoborons, these studies in methods development will introduce new transformations and strategies that will streamline synthetic chemists' approaches to complex molecules important to biomedical research.

NIH Research Projects · FY 2025 · 2021-05

PROJECT SUMMARY/ABSTRACT Cardiovascular disease (CVD) is the number one killer of Americans, and also causes significant economic costs to the nation. Thus, understanding the mechanisms through which CVD develops is of paramount importance if we are to successfully identify those at risk for CVD, and intervene to ultimately prevent CVD-related death and economic impact. Psychological stress reactivity has long been appreciated as a risk factor for negative CVD- related outcomes, and recent work suggests that inflammatory reactivity to stress is a critical biological mechanism through which stress increases risk for CVD. However, there are significant gaps in our current knowledge regarding the neural predictors and molecular pathways through which stress leads to inflammation. These knowledge gaps are critical to fill if we are to develop a full mechanistic understanding of how stress leads to CVD risk, and may also shed light on future intervention targets. Thus, the present project will use cutting- edge computational methods to identify neural signatures of stress-related inflammatory reactivity, and will use pharmacological tools to block an important stress-signaling pathway (i.e., beta-adrenergic signaling) and examine its effects on neural and inflammatory reactivity to stress. Study 1 (N=100) will use fMRI to examine neural responses to a social evaluative stress task, with blood samples taken before and after the stressor assayed for pro-inflammatory gene expression and circulating inflammatory proteins. We will use innovative multivariate machine learning analytic techniques to identify the neural patterns that predict changes in inflammation, as well as network-based analytic tools from mathematics to examine how large-scale brain networks change configuration in response to stress in ways that are linked to inflammation. In Study 2 (N=120), we will conduct a mechanistic, randomized, double-blind, placebo-controlled trial of the beta-adrenergic receptor blocker propranolol to examine how blocking beta-adrenergic signaling impacts neural and inflammatory responses to the social evaluative stress task. Together, these two studies will allow us to establish the neural signatures of stress-induced increases in inflammation (Aim 1), determine the effects of beta-adrenergic signaling on neural responses to stress (Aim 2), and examine the neural mediators of beta-adrenergic related attenuations in stress-related inflammatory reactivity (Aim 3). In doing so, this project will ultimately help identify neural signatures of risk for stress-related inflammation, as well as novel targets for future intervention to ameliorate the impact of stress on the brain and body and reduce the health and economic burden of CVD.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY – UNIVERSITY OF NORTH CAROLINA-CHAPEL HILL, FROHLICH Cognitive control requires the brain to dynamically allocate limited resources to manipulate internal representations as a function of behavioral demands, a process referred to as output-gating. Output-gating comprises two intertwined cognitive processes: the selection of relevant information and the suppression of irrelevant information. These two cognitive processes have been correlated with oscillatory neuronal network activity in two distinct frequency bands and network locations: theta oscillations (4-7 Hz) in prefrontal cortex (PFC) for selection and alpha oscillations (8-12 Hz) in posterior parietal cortex for suppression. However, the causal role of these oscillations and their interactions in output-gating has yet to be established. To address this gap, this proposal examines the causal role of theta and alpha oscillations in output-gating across multiple scales with individualized brain stimulation paradigms to provide a mechanistic delineation of how these oscillations support behavior, coordinate network activity, and regulate neuronal spiking activity. The objective of AIM 1 is to demonstrate the causal role of theta and alpha oscillations in selection and suppression, respectively. To accomplish this, theta and alpha frequency rhythmic transcranial magnetic stimulation is applied in healthy participants to frontal and parietal sites with simultaneous electroencephalography (EEG) during a working memory task with a retrospective cue that drives output-gating. The hypothesis of AIM 1 is that frontal theta activity coordinates the selection of relevant information, while parietal alpha activity coordinates the suppression of irrelevant information. The objective of AIM 2 is to spatially resolve the theta and alpha network dynamics that support selection and suppression, respectively. To achieve this objective, direct cortical stimulation combined with invasive EEG will be used in epilepsy patients with implanted electrodes for clinical purposes. The hypothesis of AIM 2 is that connectivity between frontal and parietal regions establishes oscillatory dynamics critical for selection and suppression. The objective of AIM 3 is to determine how oscillatory network dynamics regulate neuronal spiking activity. This is examined by applying theta and alpha frequency rhythmic optogenetic stimulation to frontal and parietal sites in the ferret with simultaneous electrophysiology recordings during an attentional task that modulates theta and alpha oscillations. The hypothesis of AIM 3 is that theta oscillations increase spiking and alpha oscillations decrease spiking activity. The proposed work is significant since it will provide a multi-scale mechanistic understanding of how theta and alpha oscillations coordinate output-gating. The proposed aims are innovative since they employ synergistic causal perturbations through targeted brain stimulation paradigms with concurrent electrophysiology, enabling the manipulation of oscillatory dynamics and the delineation of their role in coordinating neuronal spiking, network organization, and behavior. This work will provide the foundation for the future development of brain stimulation interventions that target impaired brain network oscillations for the restoration of cognitive deficits in psychiatric illnesses.

NIH Research Projects · FY 2025 · 2021-04

Pain is currently the most common cause of long-term disability in the United States, affecting over 70 million Americans and 1.5 billion people worldwide. The first line of treatment for pain has been opioids, however their use is associated with addiction, dependence and overdose. A strategy for reducing the reliance on opioids for treatment of chronic pain is to better understand the circuits that mediate different aspects of chronic pain and identify non-opioidergic mechanisms to restore normal circuit function and behavior. Corticotropin releasing factor (CRF) signaling in the bed nucleus of the stria terminalis (BNST) has been shown to regulate both nociceptive and affective/motivational behaviors associated with pain, however the clinical utility of pharmacologically targeting the CRF system has yet to be explored. Given the critical need to develop treatments to target affective and motivational aspects of chronic pain, this could be an important path forward. We hypothesize that persistent inflammatory pain leads to increased activation of CRF signaling in the BNST leading to disrupted affective behaviors and reduced motivation. We posit that modulators of BNST function that oppose CRF signaling could provide a novel approach to investigate and treat both nociceptive and motivational/affective aspects of pain. Relevant to this, we have found in preliminary studies, a population of periaqueductal gray dopamine (PAGDA) neurons that project to the BNST that exhibit anti-nociceptive properties when activated, highlighting the possibility that dopamine in the BNST is a critical suppressor of pain-related behaviors. Previous studies have found that these PAGDA neurons play a key role in opioid induced anti-nociception. The objective of this proposal is to rigorously and mechanistically determine the role of the PAGDA to BNSTCRF circuit in inflammatory pain-driven changes in behavior, with the long term goal of identifying new treatments for inflammatory pain. We have collected strong data showing that acute pain engages this circuit, and hypothesize that this initial engagement leads to plasticity contributing to the persistence of inflammatory pain, and the development of emotional behaviors associated with inflammatory pain. This will be accomplished via three convergent aims

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY Sleep is an essential conserved behavior seen throughout life and is critical for brain health and maintenance of cognitive functions such as learning and memory. Sleep disruption is intimately linked to aging and believed to expose individuals to risk of developing Alzheimer's Disease (AD). After AD onset, continued decline in sleep amount/quality is associated with progressive decline in memory performance and cognition. Therefore, sleep disruption is a source of vulnerability as well as a potential therapeutic target to treat disease. A detailed molecular understanding of the ontogeny of sleep disruption could aid in the development of earlier diagnosis for AD, and in the identification of a therapeutic window for sleep-based medicines. We propose that promoting quality sleep during the early stages of AD may delay or halt progressive cognitive decline. However, the molecular basis of sleep's restorative processes that support cognition is poorly understood. Neuronal synapses are the structures responsible for forming and storing memories, particularly in forebrain structures such as the hippocampus and cortex. Our previous work shows that synapses are a major target for the restorative actions of sleep. We have shown that a form of synaptic plasticity called homeostatic scaling-down is engaged in the brain during sleep to support learning and memory functions. Synapse dysfunction is also known to occur early in AD progression when the Tau protein begins to accumulate in the brain. We hypothesize that aberrant synaptic Tau induces synaptic dysfunction by altering homeostatic scaling- down, leading to hyperexcitability and sleep disruption. Sleep disruption, and loss of the restorative homeostatic scaling, then accelerates disease pathology and cognitive decline. Preliminary findings indicate sleep disruption is an early phenotype in a Tau-based mouse model of AD. In aim 1 we examine the interaction between hallmark AD pathologies, amyloid plaques and Tau tangles, in driving sleep disruption, and examine the necessity of Tau or amyloid in sleep disruption onset. We test the relationship between sleep disruption and Tau pathology to establish sleep disruption as a biomarkers of pathology. In aim 2 we will use an in vitro model system to dissect the molecular mechanisms by which pathogenic Tau proteins affect synapse function. We will examine a particular cleaved Tau species known to accumulate at the synapse in AD human brain, and examine the effect of cleaved Tau on restorative homeostatic scaling-down. In aim 3 we will examine the sleep-dependent regulation of the endocannabinoid system during aging in AD model mice. Our preliminary data show that endocannabinoid signaling is engaged during homeostatic scaling in cultured neurons, and that regulation of endocannabinoids during the sleep-wake cycle is disrupted in AD model mice. We show that acutely increasing the endocannabinoid anandamide using a pharmacological approach promotes sleep in symptomatic AD mice. We will test the therapeutic efficacy of this sleep-promoting strategy in AD mice, with the translational implications of modifying sleep behavior to alter AD onset or progression in human patients.

NIH Research Projects · FY 2025 · 2021-04

Project Summary Now in its 13th year, the Psychiatric Genomics Consortium is perhaps the most innovative and productive experiment in the history of psychiatry. The PGC unified the field and attracted a cadre of outstanding scientists (802 investigators from 157 institutions in 41 countries). PGC work has led to identification of ~500 genetic loci in the 11 psychiatric disorders we study. Our work has led to 320 papers, many in high-profile journals (Nature 3, Cell 5, Science 2, Nat Genet 27, Nat Neurosci 9, Mol Psych 37, Biol Psych 25). As summary statistics are freely available, psychiatric disorders often feature prominently in papers by non-PGC investigators. To advance discovery and impact, we propose to continue the work of the PGC across 11 disorder groups. Considerable new data are coming in the next five years. We thus can rapidly and efficiently increase our knowledge of the fundamental basis of major psychiatric disorders. Aim 1: we will continue to advance genetic discovery for severe psychiatric disorders in all working groups, systematically interface with large biobank studies to ensure maximal comparability, and aggressively promote new studies of individuals with psychiatric disorders from diverse ancestries to increase discovery and improve fine-mapping. Aim 2: most studies analyze common variation (Aim 1), rare CNV (Aim 2), and rare exome/genome resequencing results (via collaboration) in isolation: we will apply an integrative framework to rigorously evaluate the contributions of all measured types of genetic variation on risk for psychiatric disorders. Aim 3: we will move beyond classical case-control definitions to a more biologically-based and nuanced understanding by enabling large trans-diagnostic studies, convene trans-disciplinary teams to use genetics to address unresolved questions about the nature of psychiatric disorders, and to promote large studies of the severest cases seen in psychiatric practice (leveraging the global reach of PGC investigators). Aim 4: we will work to maximize the impact of our work via translational efforts: close collaborations with neuroscience consortia to understand the biological implications of our findings; work to identify modifiable causal risk factors; and work to robustly predict clinical outcomes and identify patient subsets. Aim 5: we will increase impact of our work by extending and formalizing outreach to different communities (including pharma and biotech), via digital media (Twitter, Facebook, Wikipedia), and by developing, distributing, and updating resources/educational material for patients, families, and medical professionals. We will convene a Scientific Advisory Board to ensure we respond positively to those invested in our results Successful completion of this body of work will greatly advance knowledge of the genetic basis of psychiatric disorders with potentially major nosological and treatment implications. These goals are consistent with a core mission of the NIMH, and the central idea of the PGC: to convert the family history risk factor into biologically, clinically, and therapeutically meaningful insights.

NIH Research Projects · FY 2026 · 2021-04

PROJECT SUMMARY Women in sub-Saharan Africa face an unacceptably high risk of HIV acquisition during pregnancy and breastfeeding. Daily oral pre-exposure prophylaxis (PrEP) with emtricitabine/tenofovir disoproxil fumarate (FTC/TDF) is effective in reducing HIV acquisition and is recommended in pregnancy. At standard FTC/TDF doses, however, tenofovir drug concentrations are 23-58% lower during pregnancy, raising concerns about reduced efficacy. In this study, we seek to identify and evaluate the optimal dose of FTC/TDF for daily oral PrEP in pregnancy, focusing on pharmacokinetic (PK) and safety outcomes. To accomplish our aims, we plan several key activities. Dose identification (Stage 1): We will randomize 45 pregnant women at 14-24 weeks gestation to three different FTC/TDF doses—standard dose (200mg/300mg), 150% standard dose (300mg/450mg), and 200% standard dose (400mg/600mg). Each participant will undergo three “cycles” comprising 14 days of daily oral PrEP, followed by intensive PK sampling over 24 hours. The first two cycles will occur in the second and third trimesters of pregnancy at the assigned FTC/TDF dose; the third will take place at 12 weeks postpartum and use only standard FTC/TDF. We will compare tenofovir diphosphate in peripheral blood mononuclear cells (PBMCs) in each pregnancy trimester to the postpartum control condition, using defined boundaries for bioequivalence. Preliminary safety data will also be obtained. Independent review: Findings from this initial stage will be independently reviewed by an expert, multidisciplinary Study Monitoring Committee, which will recommend an increased FTC/TDF dose (150% vs. 200% standard dose) for further study. Extended safety assessment (Stage 2): We will randomize 112 pregnant women at 14-24 weeks gestation to receive either standard vs. increased FTC/TDF doses on a daily basis, under direct observation, until time of delivery. Safety monitoring will continue through pregnancy, delivery, and the first six months postpartum. We will compare renal function, adverse events, bone mineral density, weight change/growth in women and infants, and pregnancy outcomes. We will evaluate FTC and TFV (and their metabolites) in plasma, PBMCs, red blood cells, urine, and cervicovaginal fluid. PK modeling: Using empiric study data, we will develop a PK model that estimates concentrations of FTC and TDF across multiple compartments during pregnancy. Our model will consider key factors that may influence drug concentrations (e.g., body weight, gestational age, renal function) to predict safety outcomes for lengthier exposures in pregnancy. This study will be led by an experienced team of researchers, with extensive expertise in HIV, clinical trials, pharmacology, and obstetrics. Our proposal leverages the strengths of its partnering institutions, including the robust research infrastructure at the University of Zimbabwe. Over the course of this award, we will provide key insights into the PK and safety of FTC/TDF in pregnancy. Importantly, these findings will help to optimize PrEP regimens for an important but often overlooked population: pregnant women in sub-Saharan Africa.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY Globally, over 50% of those infected with HIV are women, and annually, ~50% of all pregnancies are unplanned. Therefore, there is a critical need to promote female-controlled methods of multipurpose prevention technologies (MPTs) and delivery strategies that can be disassociated from the sex act. Injectable formulations are well tolerated by men and women, are efficacious for contraception, and have high patient acceptability and compliance1-4. Innovations recently introduced into the field of systemic PrEP are long-acting (LA) formulations of antiretrovirals (ARVs) that stably release drugs over many weeks as nano-formulations and have activity in animal models of prevention5-7 and in humans8-10. Currently, there are two different LA formulations being considered for HIV prevention: a LA form of rilpivirine (RPV/TMC278) and a LA form of cabotegavir (CAB/GSK744)9, 11-12. Injections of these LA ARVs requires a 4-week ‘lead-in’ regimen using oral cabotegravir or rilpivirine to fulfil current safety considerations as once injected these agents have detectable levels for months and the drug cannot be removed or have its clearance accelerated. Despite these advances in HIV PrEP, currently there are no LA injectable MPT formulations in development mainly because of limitations of current LA injectable formulations utilizing nanoparticle suspensions whereby two drugs cannot be combined into a single injection. In this R01 grant and building on our existing data, we propose a comprehensive evaluation of a first-in-line injectable MPT that offers durable and sustained protection from HIV transmission, high efficacy of contraception, increased user compliance, and the ability to be removed in case of unanticipated adverse events or when considering discontinuation from the LA HIV PrEP and/or contraception. We will achieve this goal by developing a liquid MPT formulation utilizing excipients that form a biodegradable depot after subcutaneous injection (in-situ forming implant (ISFI)). We propose a comprehensive evaluation of this novel drug delivery approach using a highly relevant macaque model of mucosal simian/human immunodeficiency virus (SHIV) as an invaluable preclinical tool to assess the efficacy of the ISFI against SHIV acquisition. This cutting-edge combined approach will be utilized to evaluate the scientific premise of our proposal to investigate whether sustained protection against HIV acquisition and pregnancy can be achieved using a unique and highly innovative ultra-long-acting coitally-independent MPT ISFI formulation.

NIH Research Projects · FY 2025 · 2021-04

ABSTRACT The long-term objective of the proposed work is to provide evidenced-based recommendations for minimizing metabolic adverse events, particularly weight gain, in a diverse population of people living with HIV (PLWH) by identifying and quantifying variability in drug exposure that may increase risk in patient subgroups. We will consider a broad array of modifying factors of drug exposure, including demographic characteristics, prior laboratory values, body anthropometrics, frailty phenotype, and pharmacogenomics. Our focus is on the integrase strand transfer inhibitors (INSTIs) dolutegravir (DTG) and bictegravir (BIC), as well as tenofovir alafenamide (TAF). The MWCCS includes diverse PLWH underrepresented in Phase III clinical trials and thus, in the sponsor-developed population pharmacokinetic models of the drug. In AIM 1, we will enroll diverse PLWH from four MWCCS sites to collect pharmacokinetic data and further refine the knowledge of factors that influence DTG, BIC, and TAF pharmacokinetics. Drug concentrations will be measured in blood plasma and peripheral blood mononuclear cells (TAF only) in the UNC Center for AIDS Research Clinical Pharmacology and Analytical Chemistry Laboratory (CFAR CPAC). Nonlinear mixed effects models will be used to develop a comprehensive PK model for each drug of interest. Using these models, then, in AIM 2, we will retrospectively measure drug concentrations in repository specimens (using the same methods and laboratory as AIM 1, and predict drug clearance in men and women from initiation of DTG, BIC, and/or TAF. These drug clearances, predicted for multiple visits per participant, will then be analyzed as the predictors of body weight gain, increased waist to hip ratio, and increased insulin resistance (as measured by HOMA-IR) over the course of treatment on the drug of interest, with multiple measures of drug exposure. The hypothesis is that those PLWH with higher drug exposure (via slower drug clearance) will be more likely to experience the metabolic adverse events of these drugs. Upon completion, we expect to have the underpinnings of a model-based risk estimator developed for further prospective validation.