University Of Washington

NSF Awards · FY 2024 · 2024-08

Spheres are among the simplest geometric objects, serving as building blocks for more complicated topological spaces. The homotopy groups of spheres (collections of continuous functions between spheres, considered up to certain deformations groups) hold fundamental information about maps between topological spaces and have deep connections to number theory, algebraic geometry, differential topology, and geometric topology. Despite their ease of definition, there are few effective methods to compute homotopy groups of spheres. Using equivariant technology the PI will explore the rich connections between equivariant homotopy theory and chromatic homotopy theory and use them to develop powerful new computational techniques. This research will be integrated with conference organization, and graduate students and postdocs mentoring and training. The PI will also be engaged in outreach to local middle school teachers and students. The research involves a range of projects that will leverage recent discoveries in equivariant homotopy theory to advance computations in chromatic homotopy theory. The study of Lubin-Tate theories is one of the most important areas of research in chromatic homotopy theory. In 2009, Hill—Hopkins—Ravenel's resolution of the Kervaire invariant problem elevated equivariant homotopy theory as a potent tool to drive significant progress in chromatic homotopy theory and address classical problems in geometry and topology. The projects involve exploring computational structures in the equivariant slice spectral sequences of Real bordism theories and Lubin-Tate theories at the prime 2. This endeavor extends to achieving analogous results at odd primes. To achieve these goals, the PI plans to employ new equivariant techniques, including transchromatic isomorphisms, stratification results, and the generalized Tate diagram of spectral sequences. These methods will enable extensive equivariant chromatic computations, establish general differential patterns, and reveal a broader range of transchromatic phenomena in the equivariant slice spectral sequences of norms of Real bordism theories and Lubin-Tate theories across various groups and heights. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-08

This research project will develop new theory and methods for assessing the sensitivity of causal inferences to violations of underlying assumptions. Applied causal inference work in the social, behavioral, and economic sciences often use a handful of methods commonly known as "quasi-experimental" designs. However, the validity of these methods requires strong assumptions about the data-generating process, many of which are difficult to defend in practice. What if these assumptions are false? In such cases, sensitivity analyses play an essential role by allowing researchers to quantify how strong violations of key assumptions need to be in order to substantially change a research conclusion. This project will develop a unified theoretical framework that allows researchers to easily quantify how violations of key assumptions affect causal effect estimates using such methods. These results will allow applied scientists and decision makers to draw robust and trustworthy conclusions using valid, but imperfect scientific evidence. Graduate and undergraduate students will be trained and mentored. Dissemination activities for the new methods will include redesigned academic courses, workshops on sensitivity analysis for practitioners, and the development of open-source software. This research project will develop a comprehensive suite of sensitivity analysis tools for popular identification strategies in causal inference, such as instrumental variables, difference-in-differences, and regression discontinuity designs. The investigator will bound the bias due to violations of common assumptions utilized in these settings, such as violations of the exclusion and independence restrictions in instrumental variable estimation; violations of the parallel trends assumption in differences-in-differences designs; or violations of the continuity assumption at the cutoff point in regression discontinuity designs. Analytical formulas will be derived for the bias due to such violations, as well as easily interpretable sensitivity statistics for routine reporting that can be used to quickly communicate the robustness of a result. Other innovations will include allowing for multiple, simultaneous violations of assumptions, incorporating expert knowledge on the relative importance of variables to bound the bias, and enabling valid statistical inference using both classical methods as well as leveraging modern machine learning algorithms. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-08

This project brings together U.S.-based and Bhutan-based scientists to investigate key questions on high mountain glaciers and their impact on landscape change in the Bhutanese Himalaya using geophysics. High mountain glaciers serve as important agents of erosion as well as sediment and water sources that contribute to many of the major rivers in Asia. Scientists have observed significant changes to Himalayan glaciers in response to a changing climate, which could severely impact downstream rivers and the populations that rely on these rivers for survival. Furthermore, melting glaciers can lead to glacial outburst floods, which occur when a glacial lake breaches its natural dam, resulting in a sudden release of flood water. This project will use geophysical methods including radar and seismic experiments to study the Lunana glaciers region of Bhutan, to better understand landscape change, glacier dynamics, and hazards. This project will improve our understanding of sediment transport, landscape change, glacier movements, and glacial outburst floods in the Bhutanese Himalaya. It will also test the potential of seismology to monitor sediment transport and river levels downstream of the Lunana glaciers and the Pho Chu River Valley, including the possibility for flood early warning. Seismic and radar surveys will reveal ice thickness, hydrologic processes, glacial moraine dam stability, the abundance and state of permafrost and debris-covered ice in the region, and subglacial sediments or materials. The project also aims to build geoscience, research, and field skills capacity in Bhutan and the Himalayan region. This project is jointly funded by three EAR Programs: Geomorphology and Land Use Dynamics (GLD), Hydrologic Sciences (HS), and Geophysics (PH). This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY/ABSTRACT: Stroke is not just an event but a chronic illness with tremendous impact. Every year, 5 million people die of stroke, and another 5 million are left with disabling impairments of cognitive and physical function. Progress in the acute treatment of stroke has reduced the number of patients who die but also increased the number of survivors, who along with their family members are faced with many challenges over the long term. These challenges range from cognitive and physical impairment to psychosocial and existential suffering. One third of stroke survivors develop cognitive impairment due to vascular contributions to cognitive impairment & dementia (VCID), which may worsen over time due to concomitant Alzheimer disease and related dementia (ADRD) or immunological processes triggered by the stroke itself. Post-stroke cognitive impairment reduces a patient’s ability to participate in rehabilitation and increases caregiver burden. Over 1 in 3 stroke survivors suffer advanced symptoms of pain, anxiety and depression, and these symptoms can become more frequent or intense in those who are actively dying. As families face immense grief and prognostic uncertainty, clinicians need to help them maintain hope but also understand the severity of the disease, so that they can effectively discuss treatment options across the range of possible outcomes. About 1 in 3 patients die in the hospital, most of them after a decision to withdraw life-sustaining treatments, such as mechanical ventilation, artificial nutrition, and hydration. For those who survive a severe stroke, most patients and families face a completely different reality with new social roles, high symptom burden, and frequent rehospitalization. Therefore, we urgently need interventions to support patients and families as they grapple with often conflicting realities of recovery, grief, adaptation and treatment decisions that continue for months or even years after the stroke. We propose a randomized clinical trial of a “Longitudinal Transdisciplinary Neuropalliative Care Support” (Lotus) team that follows patients and their families from early in their hospital course through six months and provides targeted support as well as helping them ensure that treatment is line with their values. This team is led by a specially trained Lotus nurse who works with a team of mental health, spiritual care, social work and neuropalliative care providers, who will support the patient and family through a combination of in-person and tele-health appointments. The team will implement complementary activities to enhance palliative care integration into stroke care: awareness, assistance, adjustment, acceptance and alignment. We hypothesize that this intervention compared to usual care will (1) improve psychological outcomes such as depression or anxiety for patients and families and (2) increase ‘hospital-free days’ for patients and improve goal concordance, meaning that patients will be more likely to receive the type of medical care that is consistent with their personal goals and preferences. Finally, we will (3) conduct a mixed methods evaluation to explore barriers and facilitators to future implementation and dissemination.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Soil-transmitted parasitic nematodes infect over a billion people around the world and can cause devastating and sometimes fatal illness. Despite this massive health burden, the biology of parasitic nematodes remains almost entirely unexplored. For example, we lack basic knowledge about the physiological specializations and sensory behaviors that enable these gastrointestinal parasites to locate and infect hosts - processes that could be targets for novel therapeutic interventions. Furthermore, virtually nothing is known about how parasitism of mammals arose, multiple times independently, from the evolutionarily conserved genomes, neural circuits, and physiology of nematodes. I have developed the potentially fatal human parasite Strongyloides stercoralis as a powerful new model to investigate the sensory adaptations of species with direct relevance to global health. In this proposal, we aim to understand how temperature cues shape the specialized behavioral and physiological responses of soil-transmitted parasitic nematodes. This proposal is a unique approach that centers the underlying biology the human parasites themselves and lies at the intersection between parasitology, neuroscience, genetics, and molecular biology. Our central hypothesis is that parasitism of mammals requires dramatic shifts in thermal behavior and physiology that arise from an evolutionarily common cluster of molecular and cellular adaptations. First, we leverage high-resolution quantitative behavioral assays to define the role of temperature in driving the parasite-specific behaviors and physiological responses of S. stercoralis across their complex life cycle, experiments we hope will establish a new platform for the development of novel therapies that block the remarkable ability of parasitic worms to locate human hosts and survive the extreme thermal environments of their bodies. Next, we will map the neural circuits underlying thermosensation in S. stercoralis, using newly optimized tools for studying neuronal function in parasitic worms. These experiments will reveal the cellular links between thermal behaviors and molecular substrates and resolve how worm neural circuits evolved to support human parasitism. Finally, we will leverage our expertise in the comparative and functional parasite genomics to identify the genetic and molecular substrates of parasite-specific thermosensory responses in Strongyloides and other parasitic nematode species; a first, critical step towards designing novel and broadly effective anti-parasitic drugs capable of disrupting infections. The results of these studies will reveal key mechanisms underlying the unique thermal adaptability of mammalian parasitic nematodes, providing an essential foundation to enable the development of novel interventional strategies. Although focused on soil-transmitted parasitic nematodes, if successful this work should provide insights into other helminths, such as schistosomes, which also actively invade mammalian hosts.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY This application is a request for funds to support a conference titled the ‘Simian Collective’ (SimCo) that highlights nonhuman primate neuroscience research in the United States and its unique role as a translational model of human neurological, neuropsychiatric diseases and disorders and healthy and impaired aging. Our collective and proposed conference seeks to build a crucial community effort to address the key challenges facing the field of nonhuman primate neuroscience in the 21st Century. The conference is strategically organized to stress that maximizing the impact of nonhuman primate neuroscience research in the modern era necessitates complementary goals of Science, Ethics, Education and Advocacy. Owing to the shared functional organization of the primate brain, our simian cousins are uniquely powerful models from which we can learn many facets of human brain function in both health and disease. This cutting-edge research must continue to grow and receive renewed investments to prosper. The meeting will also emphasize that the phylogenetic advantages of using a primate model, which shares many unique neural and cognitive characteristics with humans, for investigative research must also be balanced by the crucial neuroethical considerations that inevitably emerge because of these similarities. Furthermore, the Simian Collective will also emphasize the responsibility of the members of the field to educate our scientific colleagues and the public about the unique importance of nonhuman primate research and to advocate for its significance. The program of the meeting emphasizes the complementary relationship between these core tenets and seeks to build a community initiative to promote and ensure nonhuman primate neuroscience research in the coming years. The past two conferences were extraordinarily successful with ~200+ attendees across all levels - student, post-doctoral fellow, assistant, associate and full professors. We anticipate our community to grow, attract new talent, and remain a vibrant contributor to neuroscience research.

NSF Awards · FY 2024 · 2024-08

Bacteria are everywhere and essential for life. Bacteria in the ocean regulate global temperatures; bacteria in soil help plants grow; and bacteria in our bodies help us digest and extract nutrition from food. Bacteria change over time through evolution, but because bacteria can gain and lose genes, their evolutionary histories may be hard to determine. This project will use recent advances in mathematics to develop a new method to estimate bacterial evolutionary histories. The method will be benchmarked against existing tools for speed and accuracy. In addition, it will provide advanced training for a postdoctoral researcher from an underrepresented group and create a training workshop for specialists on bacterial evolutionary histories. Accurate estimates of bacterial and archaeal phylogenetic trees are critical for classifying novel strains, interpreting changes in microbial communities, and understanding diversification and adaptation. A major challenge in prokaryotic phylogenetics is that gene transfer and recombination events result in different genes having different evolutionary histories. This project will implement and study a novel method for averaging gene-level phylogenies to obtain an overall phylogeny. The method can average gene trees even when not all genes are shared by all organisms. The project will implement tree-averaging in software and benchmark its performance against the most widely-used bacterial phylogenetics estimators using both simulation and modern bacterial genomics datasets. To support the specific objectives of this project, a postdoctoral researcher whose intersectional identity is underrepresented in the mathematical sciences will be hired, and a bilingual Spanish-English workshop on microbial phylogenetics for the group "Women in Bioinformatics and Data Science Latin America" will be developed. The results of this project will be available at statdivlab.github.io/phylogenetics. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-08

Artificial intelligence (AI) algorithms underpin social media. Algorithms sift through a large inventory of content, deciding what appears at the top of each user's feed. These social media AI systems can shape people's beliefs, affect their well-being, and change their behaviors. These consequences accrue to individuals, but also aggregate at the societal level where the value of social media AI has been stubbornly difficult to square with the societal harms that they produce. Such issues are in part due to the individualist values embedded in how social media AI software operates, maximizing each user's individual experience–-as inferred, for example, through their likes, retweets, and surveys–-at the cost of societal preferences, such as community health and civic engagement. This project aims to shape an alternative future where social media AI software aids us in achieving societal goals, by demonstrating the feasibility of integrating such societal objectives into social media algorithms used to prioritize content in users’ feeds. The project goal is to create a method that can build translational science on top of social science and computer science, and develop engineering solutions that can be deployed at scale on social media, if desired. This project will develop techniques for encoding societal values into social media ranking algorithms. Our multi-disciplinary team of researchers aims to 1) introduce a novel method that leverages the precise language of definitions and measurements of the social science constructs to build algorithmic objective functions using large language models (LLMs), referred to as societal objective functions, which can be deployed broadly as weights in social media ranking algorithms; 2) create a pluralistic algorithmic library of such societal objective functions based on rigorous and empirically validated social science theory articulating a broad space of values; and 3) build methods to integrate multiple potentially-competing values and understand the trade-offs between them. To achieve these goals, the project will weave together social science and computer science insights. Social science research will articulate the design space of societal values at play, as well as careful definitions and measurements of each of these values. Computer science research will translate these social scientific insights into AI models that agree with community ratings on the values expressed in social media content, enabling integration into feed ranking algorithms. By conducting large-scale field experiments with diverse populations, this project will provide empirical evidence on the impact of integrating a pluralistic library of societal values into such algorithms. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/ Abstract Social justice is the primary ethical value underlying health equity. The public health literature, due to its disciplinary limitations, does not “secure” conceptions of health equity to an in-depth theory of social justice. Political philosophy offers several theories of social justice. Which of these are best suited to health equity? The objective of this solo-authored book project is to answer this question. The book has three specific aims: To identify and critically assess (1) the implicit ethical goals that underlie much public health research, education, and policy on health equity; (2) the dominant theories of social justice that are frequently applied to public health and healthcare ethics; and (3) alternative theories of social justice that could be applied to health and healthcare. As the book will be a work in normative bioethics and its primary methodology is philosophical analysis and critical assessment of concepts and arguments, it does not have a hypothesis per se but rather a central claim to be explained and defended. The central claim is that we need to develop and apply the political philosophical theory of “relational egalitarianism” to health equity. Relational egalitarianism is a theory committed to elucidating and defending relational equality. According to this form of equality, foremost, social justice means that people must be able to stand in front of each other as equals. Health inequity, according to this theory, occurs primarily when health disparities are caused by or lead to relational inequalities, or both. The innovation of the project is that it will (1) develop relational egalitarianism to apply to health equity as an alternative to dominant analytic theories of social justice, and (2) it will engage in literature from analytic political philosophy along with theories of structural and interactional racism, sexism, transoppression and ableism from the critical theory literature. Its significance is that it will demonstrate how relational egalitarianism can help guide choices about how health equity should be understood, measured, and represented in the health sciences, policy, and education. As the reduction of health inequities, which will improve the health of marginalized groups, is frequently cited as a primary goal of public health, the book is of clear

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract Alzheimer’s disease and related dementias (ADRDs) are highly prevalent, devastating to patients and their loved ones, and encompass a diversity of etiologies and pathologies. Traumatic brain injury (TBI) and repetitive head impacts (RHI) can contribute to the initiation and/or acceleration of ADRDs, including chronic traumatic encephalopathy (CTE), but critical gaps in knowledge prevent the development of effective diagnostic strategies and therapeutic interventions. Most importantly, mechanisms through which diverse patterns of head trauma increase risk for AD/ADRD/CTE are not known, nor is it clear how multiple pathological processes interact to influence symptom manifestation and disease progression. These knowledge gaps must be addressed to enable in vivo diagnosis and disease-modifying therapeutics in our lifetime - the highest-priority recommendation for post-TBI AD/ADRD from the 2022 ADRD Summit. To address these critical issues, we propose NEW-HOPE-TBI, a unique and powerful cohort and open-access biospecimen, imaging, and data resource to enable and enhance research to better understand connections between macroscopic (neuroimaging) and microscopic (neuropathology) features of posttraumatic AD/ADRD/CTE. To accomplish this, we propose to expand our existing nationwide network of well-characterized brain donors from diverse cohorts with heterogeneous TBI exposures and/or AD/ADRD (Aim 1), broadly characterize co-pathology burden about head trauma (HT) exposure patterns (Aim 2), identify quantitative neuropathology and imaging signatures of cognitive and behavioral decline in chronic TBI and post-traumatic neurodegeneration (Aim 3), and deeply phenotype meso- to micro- scale vascular pathology in TBI (Aim 4). Through this work, we will build a unique open-access biospecimen and data biorepository that includes digital pathology and imaging data linked to accessible brain tissues and biofluids, and critical metadata including TBI exposure history, cognitive and behavioral health metrics, and social determinants of health, from well-characterized research donors (Aim 5). We have assembled a world-class team of scientists, clinicians, neuroimagers, neuropathologists, and biostatisticians to fulfill the goals of this proposal, including Dr. Tim Brown, a bioethics expert who is committed to advising NEW-HOPE-TBI concerning diversity, equity, and inclusion in our research team and cohort. Finally, to promote AD and ADRD neuropathology research far into the future, we place great emphasis on the career development and engagement of junior neuropathologists and fellows who represent the next generation of post-traumatic neurodegenerative neuropathology researchers. The proposed work is made possible by leveraging clinical TBI and AD/ADRD studies, network-based autopsy with ex vivo MRI, postmortem data that far exceed the NIH CDEs, a world-class team of senior, mid-level, and early-stage investigators, and a unique focus on neuroethics to enhance diversity in neuropathology research. No other team in the world can lead the science and data sharing proposed herein.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Antiviral systems are critical to the survival of their host. They protect from virally induced morbidity and mortality across all known domains of life and play an essential role in controlling the endogenized viral elements contained in genomes, the unchecked expression of which can lead to debilitating autoimmune diseases in humans and developmental problems in multicellular organisms. Many antiviral systems encode the exquisite ability to detect and modify or destroy specific nucleic acids, and this has made those components transformative tools for biotechnology and medicine. The directed discovery of antiviral systems in prokaryotes has exploded the number of antiviral systems that were known and reshaped how we think about the evolution of antiviral immune strategies. In eukaryotes, far less discovery research has been done even though there is significant lack of conservation between the well-studied antiviral systems of humans, plants, and insects with those of microbial eukaryotes such as amoeba. Not only are microbial eukaryotes the most abundant eukaryote, but they are also often hosts of the most enigmatic viruses discovered to date, the Megavirales family; giant viruses whose physical and genome size can rival that of bacteria. The machinery these microbial hosts use to control viral infection, and how these viruses subvert these responses remains almost entirely unknown. This proposal aims to address this unanswered question through first understanding how Acanthamoeba spp. defend themselves against members of the Megavirales family before expanding that understanding (and the tools used to arrive at it) to other microbial eukaryotes. We will combine principles of the virus-host arms race with cutting- edge AI-driven protein structure prediction and homology detection to uncover viral proteins that are involved in antagonizing currently known pathways of viral restriction in Acanthamoeba. We will further use environmental sampling to establish a collection of viruses and wild amoeba isolates that display a range of susceptibilities to our giant viruses to identify new systems of restriction within the Acanthamoeba genus. Finally, we will combine our environmental sampling with discovery-based functional metatranscriptomics to realize the hidden antiviral capacity of the abundant microbial eukaryotes. Ultimately, these discoveries will lead to the first comprehensive assessment of non-metazoan antiviral systems that are present in eukaryotes. This will not only dramatically expand our knowledge and challenge ideas of how eukaryotic antiviral systems evolved, but the results of this proposal will provide new molecules that can be used to better human life and life quality through their technological and direct biomedical applications, as have so many antiviral systems before.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Diarrhea caused by enteric pathogens infections is a leading cause of morbidity and mortality for children in low- and middle-income settings. Upgrading drinking water infrastructure is a common strategy for reducing enteric pathogen infections, which are predominantly transmitted through environmental media and most of which lack effective vaccines. However, upgraded water systems alone may not be sufficient to reduce exposures to enteric pathogens due to the potential for the introduction of contamination from unhygienic household water handling practices, after water is collected from the source. Seasonal changes in water availability may also influence waterborne disease transmission pathways, creating another potential barrier to reducing enteric disease. Here I propose to assess the impact of water handling practices and seasonality on waterborne transmission of enteric pathogens. I will analyze data from household surveys and microbiological data from drinking water alongside enteropathogen infection data from an impact evaluation assessing acute and chronic health outcomes in children up to 12 months of age associated with an upgraded drinking water system in Beira, Mozambique. A key advantage to the data from the ongoing study is the availability of microbiological data on infections with specific enteric pathogens, which may provide a more accurate estimate of health burden compared to reported symptoms alone. In Specific Aim 1, I will assess the impact of water handling practices, including water storage, household water treatment, and use of alternate water sources, on enteric pathogen infections in 12-month-old children. In Specific Aim 2, I will assess if and how household water handling practices interfere with improvements to drinking water source infrastructure using a mediation analysis framework. Specific Aim 3 will then assess the role of seasonality on differences in enteric health outcomes and drinking water exposure pathways. Findings from this analysis will provide insight into how behavioral and seasonal factors impact the relationship between drinking water quality and enteric pathogen infections in a low- income, urban setting. Understanding these factors will allow for insight into drivers of enteric pathogen transmission and potential targets for addressing the high burden of enteric disease in settings with limited infrastructure and vaccine availability.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY – OVERALL: MACHINE LEARNING-ENABLED DESIGN OF PROTOTYPE PATHOGEN VACCINES AND ANTIBODIES We propose a highly synergistic Center focused on developing end-to-end strategies for pandemic preparedness vaccine development for bunyaviruses and paramyxoviruses. Our Center brings together five research institutions with complementary and synergistic expertise in computational protein design, structure-based vaccine design, mRNA vaccines, structural biology, viral entry, viral diversity and evolution, animal model development, high biosafety-level containment virology, vaccinology, and vaccine process development and technology transfer. Our team has real-world experience in vaccine and biologics product development in both academic and industry settings. Our Center comprises five Scientific Projects supported by three Scientific Cores, an Administrative Core, and a Data Management Core. Our Scientific Projects include: 1) Development of computational methods for vaccine and biologics design, 2) Fundamental research on viral entry and receptors, 3) Antigen design, 4) Protein nanoparticle vaccine development, and 5) mRNA vaccine development. We will structure our efforts in two phases: in Phase 1 (Years 1-3) we will focus on developing vaccines for our prototype pathogens and in Phase 2 (Years 4-5) we will apply those learnings to two new bunyaviruses and two new paramyxoviruses to demonstrate that our computational and experimental approaches generalize across viral families. Our prototype pathogens are: Lassa virus (LASV; arenaviruses), Rift Valley fever virus (RVFV; phenuiviruses), and Hendra virus (HeV; paramyxoviruses). We carefully selected our prototypes as we believe they present specific vaccine design challenges which, if we are successful in solving, will facilitate the development of vaccines against related viruses. Simultaneously, the antigens from viruses in these three families have some similarities that will give rise to synergies in our design approaches. The structure of our Center will allow maximal synergy between our groups in pursuit of its central output: to define generalizable approaches and tools to develop vaccines and biologics for emerging pathogens with pandemic potential.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Protein phosphorylation is the most important and most extensively studied mechanism by which cells rapidly sense signals, transduce signals, and execute decisions. Protein phosphorylation provides a means to dynamically regulate protein function, is integral to all cellular processes, and abnormal phosphorylation is associated with many diseases. In our previous research, we developed and applied experimental and computational phosphoproteomics methods to systematically interrogate the signaling network and reveal its architecture and design principles. Our rigorous research delivered reproducible methods, which are essential to analyze phosphoproteomes in multi-perturbation studies. Our applications delivered knowledge that is crucially important to understand why different cell types respond differently to the same stimuli to accomplish different functions. Building on the methods we developed and knowledge acquired, we propose to focus our research for the next 5 years on the functional aspects of protein phosphorylation, addressing the outstanding question of how phosphorylation regulates the organization of the cellular proteome. We will expand our studies in multiple directions that represent the next frontier in signaling biology. First, we will identify phosphosites that are relevant to protein functions at the proteomic scale. Second, we will study how phosphorylation regulates the multi-level organization of the cellular proteome from transient protein interactions to subcellular organelles. Third, we will study the differences in signaling of morphologically distinct cellular phenotypes. Fourth, we will develop methods to include phosphorylation of tyrosine and histidine in phosphoproteomic studies. Fifth, we will assess the impact of mutations on protein functions at the proteomic scale. To achieve these goals, we will combine mass spectrometry-based proteomics, high throughput biochemistry, molecular biology, advanced microscopy, cell sorting, and computational and statistical methods, placing a strong emphasis on expanding the capabilities of current proteomic methods. This research will deliver new methods and a wealth of knowledge of protein phosphorylation in a spatial, temporal, and functional context; which can be expanded in many new directions. Our discoveries on the basic functioning and regulation of proteins can have an immediate translational impact by informing us about the functional consequences of mutations and changes in phosphorylation in disease states.

NIH Research Projects · FY 2024 · 2024-08

Project Summary/Abstract “Necroptosis” is a form of cell death with roles in host defense, autoimmunity, and cancer. Necroptosis is associated with inflammation and immunity, but how cells in which the necroptotic pathway is active alter the immune response is poorly understood. Much of the prior research in this area has focused on the role of lytic cell death and the release of “DAMP” molecules in necroptosis-induced inflammation. However, recent work from our group and others has highlighted key roles for proteins of the necroptotic pathway—including RIPK1 and TRIF—in activating inflammatory transcription programs, and has demonstrated that activation of “necroptosis” can drive cytokine production in the absence of cell death in some settings. The work proposed here will test the hypothesis that transcriptional signaling, not lytic cell death, represents the most immunologically significant output of the “necroptotic” pathway, and that necroptotic cell death may actually reduce inflammation by eliminating cells in which inflammatory transcription programs are active. To do this, we will focus on thee Aims: First, we will define the interactions and transcription programs activated by necroptotic pathway components in cultured cells, using engineered proteins and natural ligands. Second, we will study the role of necroptosis pathway activation in vivo, by delivering engineered, constitutively active forms of necroptosis inducers to the lung epithelium of mice. Third, we will evaluate the roles of necroptotic transcription and cell death in the immune response to influenza A virus, a potent activator of the necroptotic pathway. The work proposed here will use novel experimental tools to understand the immune response to “necroptosis” in vivo. Given the emerging roles of this pathway in infection, autoimmunity and cancer, and the substantial efforts underway to target it therapeutically, we suggest that these studies are timely and potentially impactful.

NIH Research Projects · FY 2025 · 2024-08

SUMMARY/ABSTRACT During the past decade in Latin America, new yearly HIV infections rose 21%, and as of 2022, HIV infections were still rising. Of the 3.8 million people living with HIV in the Americas, 2.8 million live in Latin America. The HIV epidemic continues to be highly concentrated among gay, bisexual, and other men who have sex with men (GBM), especially in Mesoamerican countries such as Guatemala (9%), Belize (13.9%), Mexico (14.9%) and El Salvador (16.3%). Research points to multiple co-occurring psychosocial and structural conditions, including HIV- and sexuality-related stigmas that act as intertwined forces that potentiate HIV transmission among GBM. This study addresses intersectional stigmas experienced by Indigenous GBM (IGBM) in Guatemala. This study will adapt a patient-provider stigma-reduction intervention - Finding Respect and Ending Stigma around HIV (FRESH). FRESH is workshop-based intervention that has been employed to reduce stigmas among healthcare workers and GBM around the world, including a recent Spanish-language version in the Dominican Republic. Guatemala is a diverse society in which close to 50% of the population identifies as Indigenous. Indigenous Guatemalans who also identify as GBM, experience intersectional stigmas, including racial discrimination, which increase vulnerability to HIV. Due to a crumbling public health system, and discrimination towards Indigenous people at public hospitals, Indigenous traditional healers (ITH) are the first line of response to those seeking health services. The adapted FRESH intervention, named KABAWIL in Maya K’iche language will be implemented with IGBM and will include, for the first time, ITH. Our aims include: Aim 1: We will use the ADAPT- ITT framework to adapt FRESH and produce the culturally tailored KABAWIL intervention. We will conduct in- depth interviews (20 per group) and two focus groups with IGBM and ITH. Aim 2: We will use a randomized wait- list control trial design to pilot test the intervention with 120 participants. Thirty IGBM and 30 ITH will be randomly assigned to three KABAWIL intervention workshops (10 GBM + 10 ITH per workshop; n=60). The other 30 GBM and 30 ITH will be assigned to the 3-month wait-list control. We will assess the intervention’s preliminary efficacy on increasing HIV testing, PrEP uptake, PrEP/ART adherence, and decreasing experiences of stigma and discrimination. Aim 3: Evaluate facilitators and barriers to the implementation of the KABAWIL intervention. We will conduct a post-implementation, mixed-methods assessment guided by the Consolidated Framework for Implementation Research (CFIR). We will conduct interviews with IGBM (n=10), ITH (n=10), and interventionists (n=10) to identify contextual and organizational factors that may impact the feasibility and acceptability of the intervention, and determine organizational/contextual fit for the design of a larger hybrid effectiveness implementation trial to establish KABAWIL as a model for reducing intersectional stigmas and improve HIV prevention and care for indigenous people throughout the Americas.

NIH Research Projects · FY 2025 · 2024-08

Theranostics is a type of precision medicine in which targeted radiolabeled compounds are being used for both diagnosis and treatment. Targeted therapies can increase therapeutic effectiveness while limiting side effects. Since the publication of the NETTER1 trial results (1,2), the use of 177Lu-DOTATATE to treat neuroendocrine tumor (NET) patients has increased significantly. The protocol for the trail was four 200 mCi administered treatments at 8-week intervals. This protocol received FDA approval in the United States and has become the standard of care (SOC) for most sites throughout the world. On the other hand, there is a large group of physicians and scientists that believe 177Lu-DOTATATE treatments should be personalized for each patient. 177Lu emits electrons for therapy, but it also emits gammas that can be used for imaging. Therefore, it is possible to quantitate dose to a person’s organs at risk (OAR) and tumors using imaging techniques, such as SPECT/CT. Assessing OAR dosimetry after each treatment allows for personalization of 177Lu-DOTATATE therapy. Retrospective studies have demonstrated significant lengthening of progression free survival (33 vs. 15 mos) and overall survival (54 vs. 25 mos) when patients were able to continue treatments up to 23 Gy delivered dose to their kidneys (4). In addition to OAR dosimetry, there is growing interest in personalizing therapies based upon tumor biology. Thus, the goal of this work is to develop and validate a cost effective, patient friendly technology that will enable personalization of 177Lu-DOTATATE using daily measurements of the radioactivity in the patient’s OAR and tumors without requiring serial visits to an imaging center. The Personalized Remote Radiation Tracking Portable Organ Dosimetry Device, PRRT PODD or PODD for short, is a patient-friendly radiation measuring device that the patient can use in their homes. The primary OAR is the patient’s kidney; however, the PODD will also provide data for accurate dose estimation of other OAR and physician selected tumors, thus enabling treatment personalization based upon maximum tolerated dose to OAR, cumulative tumor dose, therapeutic ratio (i.e., ratio of tumor dose to dose to OAR), or other dosimetry-based metrics. Our project consists of four specific aims: 1) Design, develop, and characterize compact imaging detectors and associated software tools for use with the PODD; 2) Use realistic, digital NET patient phantoms and simulation tools to optimize PODD design for both OAR and tumor dosimetry; 3) In clinic patient studies to validate quantitative organ and tumor measurement capabilities of PODD; and 4) Conduct a patient study including at home measurements comparing OAR and tumor dosimetry estimates using PODD to serial SPECT/CT imaging. At the conclusion of this project, we will have validated our PRRT PODD technology on a 36-patient cohort and the technology will be ready to support clinical trials to study the benefits to patients of personalized 177Lu-DOTATATE therapies, where personalization can be based on dose to patients’ OAR or tumors and on tumor biology.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Infectious gastroenteritis is a leading cause of morbidity and mortality worldwide. Diarrheagenic E. coli (DEC), particularly enteropathogenic (EPEC) and enteroaggregative (EAEC) E. coli, are well established as important causes of gastroenteritis in infants living in low- and middle-income countries (LMICs). Although some observations suggest that adults in LMICs may asymptomatically harbor DEC and that DEC may be a frequent but unappreciated cause of diarrhea in young children in HICs, the contribution of these organisms to gastrointestinal (GI) illness in adults living in high-income countries (HICs) is poorly understood. Multiplex PCR panels enabling rapid detection of the common causes of infectious gastroenteritis, including EPEC/EAEC, were approved by the FDA in 2014, and are increasingly used by clinical laboratories. Recent studies employing multiplex panels indicate that DEC are commonly found in adults in HICs. A multicenter study of patients with gastroenteritis in 10 European countries found that EPEC was the most common pathogen detected. However, EPEC/EAEC were frequently found in samples containing multiple pathogens, raising questions about their clinical significance. In our laboratories at the University of Washington, EPEC and EAEC comprise 22.5% and 12.3% of all positive stool panel results, respectively. In this proposal, I plan to perform both retrospective and prospective studies to better define the clinical significance of EPEC/EAEC in U.S. adults. The specific aims are to: 1) perform a retrospective analysis of the clinical features, risk factors, antimicrobial susceptibility, and outcomes of symptomatic adult patients with stool samples positive for EPEC/EAEC, and 2) create a repository of EPEC/EAEC strains from symptomatic adult patients using prospectively collected residual stool samples and perform whole genome sequencing (WGS) to define the molecular epidemiology of DEC in Seattle adults. These studies will provide new insights into the clinical importance of DEC in HICs and inform strategies for their management and prevention. The research will be supplemented by coursework and specialized training activities as part of a five-year career development plan to allow me to become an independent investigator. Intensive training in next-generation sequencing technologies and bioinformatics with mentoring by experts in the fields of infectious diseases and bacterial genomics will allow me to achieve my long-term objective of becoming a productive independent investigator with a focus on the molecular epidemiology of bacterial infections.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Metabolic-associated fatty liver disease (MAFLD) is a fatty liver disease with dysregulated metabolic phenotype but without excessive alcohol consumption and is a significant public health concern worldwide. The pathogenesis of MAFLD involve multiple cell types and usually accompanied by decreased metabolic capacity as well as increased inflammation and oxidative stress in the liver. The intestinal environment (i.e., gut microbiome, host intestinal cell types) also contributes to MAFLD via the gut-liver axis. MAFLD has traditionally been linked to nutritional imbalances, such as a western diet (WD). However, the pathogenesis of complex metabolic disorders, such as MAFLD, is rarely caused by only one risk factor. Early life exposure to environmental stressors is an emerging contributor to the delayed onset of metabolic diseases later in life. Among various environmental stressors, epidemiological and animal studies showed that legacy and current-use flame retardants (i.e., polybrominated diphenyl ethers [PBDEs] and tetrabromobisphenol A [TBBPA]) are associated with altered carbohydrate and lipid metabolism, which are hallmarks of MAFLD. PBDEs and TBBPA can activate important xenobiotic-sensing nuclear receptors, namely the pregnane X receptor (PXR) and constitutive androstane receptor (CAR) in the liver and intestine. PBDEs and TBBPA are enriched in breast milk and can cross the placenta making neonates especially susceptible to flame retardant-induced toxicities. I demonstrated that neonatal exposure to BDE-99 (i.e., human breast milk enriched PBDE congener) persistently up-regulated proinflammation- but down-regulated lipid metabolism-related genes in mouse livers, which was accompanied by a dysbiotic gut microbiome at young adulthood. Furthermore, I observed immune cell infiltration in the liver with compromised xenobiotic and lipid metabolism pathways in hepatocytes at later adulthood from neonatal exposure to BDE-99. Large intestinal microbiota transplantation using donors that were neonatally exposed to BDE-99 showed altered the immunological landscape of the gut environment towards proinflammation in the germ-free recipients, suggesting the involvement of the gut microbiome in the dysregulated gut-liver axis later in life. Building on these findings, my central hypothesis is that early life exposure to legacy and current use flame retardants predisposes MAFLD development later in life, modulated by the gut microbiome. Specifically, the flame retardant-induced proinflammatory gut environment leads to the exacerbation of liver injuries later in life by. I will use a novel humanized transgenic mouse model with human PXR and CAR as well as their targeted human CYP3A genes to test my hypothesis with 2 specific aims: 1) early life PBDE or TBBPA exposure exacerbates MAFLD following WD; 2) altered gut environment from early life toxicant exposure critically regulates aggravation of WD induced MAFLD. The proposed work lay the foundation for how the gut microbiome, environmental stressors, and secondary risk factors interact in complex diseases.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Sensitive Treponema pallidum genome recovery through tiling amplicon sequencing Genomic epidemiology is now a standard part of the public health response to outbreaks of bacterial pathogens. For most bacteria, genome recovery is performed via shotgun sequencing of clinical isolates. Genome recovery for Treponema pallidum, the causative agent of syphilis, is more complicated due to the lack of routine culture for the organism in clinical labs. Expensive reagents and complicated protocols are required to perform hybridization capture, which is not standard in clinical or public health microbiology laboratories. Even then, the analytical sensitivity of hybridization capture does not meet the low levels of T. pallidum DNA present in most clinical samples, leading to failure of complete genome recovery in over half of clinical specimens that test PCR positive for T. pallidum DNA. Here, we propose to optimize methods to recover T. pallidum genomes from diverse specimens to allow for greater analytical sensitivity and scale. Specifically, based on recent data from hundreds of newly sequenced T. pallidum genomes, we will design a tiling amplicon sequencing approach for T. pallidum genome recovery, using 2- 3kb sized PCR amplicons compatible with both long-read and short-read sequencing. We will then fully validate this tiling amplicon library generation approach using both cultured isolates and diverse clinical T. pallidum specimens taken from across the world to determine its analytical sensitivity, specificity, precision, and accuracy. Finally, we will widely share our standard operation procedures and reagents for executing this work in research and public health laboratories across the world. The proposed work will extend the analytical sensitivity, ease of workflow, and cost-effectiveness for T. pallidum genome recovery, enabling genomic epidemiology to inform outbreak response and prevention for syphilis.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT HIV is a chronic infection requiring long-term antiretroviral therapy (ART) to halt viral replication and prevent disease progression. Despite highly effective ART, issues remain with medication access, adherence, and tolerability. When ART is stopped, the virus rebounds from a reservoir of long-lived infected cells with resumption of disease progression. Exceptions to this natural disease course are “Elite Controllers” who spontaneously suppress viral replication by highly functional HIV-specific T-cells. To achieve the robust immune responses observed in Elite Controllers, we have generated HIV-targeting chimeric antigen receptor (CD4CAR) T cells that redirect the specificity of T cells to recognize highly conserved regions of HIV. Using our validated non-human primate (NHP) model of chronic HIV infection, we have shown that CD4CAR T cells induce long- term viral remission following ART interruption in a subset of NHPs. Our data suggest that long-term CD4CAR T-cell responses will be essential to advance this therapy broadly for people living with HIV (PLWH). We are currently investigating strategies to extend CD4CAR T-cell persistence in vivo. Unlike in cancer, CAR T cells for HIV must overcome abundant soluble antigen in the form of circulating virus. Our preliminary data highlight how soluble antigen can inhibit CD4CAR T-cell killing of target cells and promote a dysfunctional immunophenotype. We hypothesize that soluble antigen must be overcome to achieve comparable success as observed for cancer. In this proposal, we will thoroughly characterize and overcome the effect of soluble antigen on CD4CAR T-cell function using our new ex vivo models of viremia to validate next-generation CD4CAR constructs. We will also extend the persistence of CD4CAR T cells using the immunostimulatory IL15 cytokine, which can boost HIV-specific endogenous T-cell and cancer-specific CAR T-cell responses. Our preliminary data confirm the potential for IL15 to enhance the cytotoxicity and survival of CD4CAR T cells. In this proposal, we will compare a series of IL15-armored CD4CAR T-cell strategies to enhance long-term persistence and durable cytotoxicity. Finally, we will combine our strategies to generate potent armored CD4CAR T cells with improved function and persistence during viremia, which we will validate and test in our NHP model of chronic HIV infection. Through the mentored research experiences outlined in this K08 proposal, Dr. Bui will develop expertise in gene therapy and adoptive cellular therapies for the treatment of HIV, which is aligned with his future career goal to become an independent physician-scientist advancing immunotherapies for infectious diseases. The hosting institutions have vibrant scientific communities in adoptive cellular therapies and infectious diseases that Dr. Bui will engage in to foster learning and build collaborations. Dr. Bui will receive comprehensive training in technical skills, scientific inquiry, and scientific communications, and will present his discoveries at conferences and in written publications. With the guidance of his expert mentors and committed support from the hosting institutions, Dr. Bui will smoothly transition to a principal investigator position by the end of his K08 support.

NIH Research Projects · FY 2026 · 2024-08

Project Summary Hypertrophic cardiomyopathy (HCM) is a heritable hypertrophy of the left ventricle that can present as sudden cardiac death and heart failure. Autosomal dominant missense variants in myosin heavy chain 7 (MYH7) can cause up to 40% of the familial HCM cases. While accurate interpretation of MYH7 variants can help identify patients at risk for developing HCM, there are currently ~1,800 MYH7 missense variants in ClinVar that are variants of unknown significance (VUS), creating diagnostic challenges for clinicians and emotional distress for patients. Furthermore, because clinical guidelines now recommend that all HCM patients undergo genetic testing, the number of MYH7 VUS is growing rapidly. Deep mutational scanning (DMS) can help address the VUS problem by determining the functional consequence of thousands of genetic variants in a single experiment. By generating accurate variant effect maps that define the function of nearly all possible missense variants in a sequence of interest, DMS can assist with clinical variant interpretation at scale. So far, deep mutational scans have been primarily practiced in cancer-derived cell lines, which are not helpful for assessing genes such as MYH7 that are only expressed in specialized cells like cardiomyocytes. Human induced pluripotent stem cells (hiPSCs) can differentiate to many cell types, offering a solution to this problem; however, gene-editing in hiPSCs is generally low throughput. We recently developed a novel gene-editing technique called CRISPR activation On-Target Editing Retrieval (CRaTER) that can edit hiPSCs at scale, enabling the generation of large variant libraries in hiPSCs for the first time. To address the growing MYH7 VUS problem, we plan to perform a deep mutational scan of MYH7 in physiologically-relevant hiPSC-derived cardiomyocytes (hiPSC-CMs). In Aim 1, we propose to generate a single nucleotide variant (SNV) library of nearly all possible MYH7 SNVs in hiPSC-CMs. We will leverage CRaTER-assisted genome-editing to be able to generate a hiPSC SNV library at this scale. In Aim 2 we will optimize and validate phenotypic assays to be able to discriminate hiPSC-CMs expressing pathogenic variants from hiPSC-CMs expressing benign variants. These phenotypic assays will inform a deep mutational scan of MYH7 that will determine the functional effect of nearly all possible MYH7 SNVs and assist with clinical variant interpretation. In summary, we will combine CRaTER, a novel gene-editing technique we developed, with hiPSCs to perform a DMS of MYH7 in genome- edited hiPSC-CMs. The results will improve the interpretation of ~1,800 MYH7 VUS, enable the interpretation of thousands of possible MYH7 SNVs that may be encountered in the future, and provide a framework by which deep mutational scans can be performed in hiPSC-derivatives for the first time, which we believe will be transformative for the field of variant interpretation.

NIH Research Projects · FY 2024 · 2024-08

ABSTRACT In several high-income nations, including the United States, infectious syphilis has been resurgent for over two decades now, while syphilis is still endemic in low- and middle-income countries. Syphilis is therefore still a public global health concern, particularly in consideration that it can lead to neurological sequelae such as dementia and stroke-like syndrome, as well as cardiovascular manifestations potentially leading to death. Furthermore, every year, about half a million pregnancies are adversely affected by congenital transmission of the pathogen. The partial success of recent syphilis control campaigns promoted by the CDC and WHO clearly highlights the necessity of devising novel ways to control this serious infection. Improving our understanding of syphilis pathogenesis and the virulence factors that allow the syphilis agent, Treponema pallidum subsp. pallidum (T. pallidum) to establish infection and persist in the host despite a robust immune response might be the key to new control strategies. However, two significant obstacles have hindered our ability to unravel the complexities of syphilis pathogenesis since the first T. pallidum strain was isolated in 1912. These barriers were the inability to propagate T. pallidum in vitro and, consequently, to genetically manipulate this difficult pathogen. In 2018, the “in vitro propagation” barrier was overcome by the discovery that T. pallidum could be propagated in a cell culture-based system. In 2021, for the first time, we overcame the “genetic manipulation” barrier and derived a T. pallidum knock-out (KO) isolate, in which a functional kanamycin resistance (kanR) cassette was used to successfully replace the non-essential T. pallidum tprA (tp0009) locus through homologous recombination after transforming the syphilis agent with a suicide vector. This newly found ability to genetically alter T. pallidum will allow us to pinpoint more clearly the role of putative virulence factors of this pathogen during infection. Here, by using an array of newly generated T. pallidum KO strains lacking critical components of the pathogen`s antigenic variation system, we propose to study the contribution of antigenic variation to T. pallidum ability for immune evasion and persistence during infection. If successful, these studies will provide our research community with an array of T. pallidum mutants to be used in comparative studies. Furthermore, our results will help settle ongoing controversies surrounding the function of the T. pallidum TprK virulence factors that originated over the last two decades due to working with a difficult pathogen that, until now, could not be genetically engineered.

NIH Research Projects · FY 2024 · 2024-08

Project Summary An emerging model of obesity pathogenesis posits a central role for the inflammatory activation of hypothalamic microglia localized to the arcuate nucleus (ARC, a key brain area for the control of food intake and body weight) in the pathogenesis of diet-induced obesity (DIO), a finding observed in mammalian species ranging from rodents to humans. While microglia activation in the ARC is a known determinant of weight gain in high fat diet (HFD)-fed mice, it paradoxically improves glucose tolerance in DIO, but the mechanisms underlying both of these metabolic effects remain unclear. Herein, we report the novel finding that in mice, DIO induces loss of specialized extracellular matrix (ECM) structures known as perineuronal nets (PNNs) in the same brain area where reactive gliosis occurs. PNNs can powerfully influence the activity of neurons that they enmesh, and in the ARC, a large proportion of AgRP and a subset of POMC neurons are among those enmeshed by PNNs. These neurons are central regulators of energy homeostasis, and their altered function in DIO is strongly implicated in obesity pathogenesis and glucose regulation. Importantly, ablating or silencing microglia reduces Npy and AgRP levels and increases POMC neuron excitability, suggesting a link between microglial activation and ARC neuronal function that promotes weight gain. Furthermore, DIO is associated with loss of hypothalamic PNNs in proportion to the degree of microglial activation, with PNN stability enhanced by interventions that ablate or limit the inflammatory capacity of microglia. Finally, removal of hypothalamic PNNs experimentally causes gliosis, hyperphagia and rapid weight gain with preserved glucose tolerance in rodents, providing strong evidence that microglia and PNNs are linked in a critical mechanism that underlies obesity pathogenesis. Here, we investigate the inter-related hypotheses that obesity-associated microglial activation induces loss of PNN enmeshment of ARC neurons, thereby altering AgRP and Pomc neuron function in ways that promote excess fat accumulation but maintain glucose tolerance. Proposed studies will first quantify the role played by microglia to regulate ARC PNN turnover. We will then 1) determine whether the effect of DIO to induce loss of ARC PNNs depends on microglial activation, 2) identify the roles of both microglial activation and PNN loss in obesity-associated dysfunction of AgRP and POMC neurons, and 3) determine the bidirectional contributions linking ARC PNN loss and microglial activation to DIO susceptibility.

NIH Research Projects · FY 2025 · 2024-08

SUMMARY/ABSTRACT The burden of noncommunicable diseases (NCDs), including cardiometabolic disease, is increasing globally, particularly in low- and middle-income countries (LMICs). NCDs are responsible for an estimated 35% of all deaths in sub-Saharan Africa, and in Mozambique nearly 60% of all DALYs are attributable to NCDs and injury. The prevalence of hypertension (HTN) and type II diabetes (T2D) are on the rise, from 33.1% in 2005 to 38.9% in 2015 for HTN and 2.9% to 7.4% for T2D across the same period. Effective strategies to meet the growing disease burden that can be integrated into – and scaled through – health systems and disease-specific platforms, like those for HIV, are needed. Similarly, little information exists on effective, population-based primordial prevention of cardiometabolic in LMIC community-based settings, including efforts to intervene in the relationship between an individual's environment, health behaviors, and risk factors for HTN and T2D. In recent years, several implementation research studies have applied the Systems Analysis and Improvement Approach (SAIA) to optimize care cascades for preventing mother-to-child HIV transmission (R01HD0757; PI: Sherr), addressing HTN among people with HIV (SAIA-HTN, R01HL142412, PI: Gimbel; SCALE SAIA-HTN, 1UG3HL156390- 01/UH3HL156390,PI: Mocumbi/Gimbel), and to pilot T2D care optimization within facilities where SAIA-HTN has been implemented, coupled with food environment assessments and Citizen Science-based nutritional education modules in three institutional cafeterias. This F31 proposal will leverage the data gathered under these studies to conduct formative research to characterize determinants for effective implementation of integrated care for T2D, HTN, and HIV embedded within ongoing health systems strengthening efforts in public sector facilities in Mozambique. We will also conduct pilot research to determine possible associations between community-based participatory education modules and changes in diet. We will evaluate barriers, facilitators, and organizational readiness to deliver medical services for T2D alongside integrated HTN and HIV services (Aim 1), leveraging qualitative data gathered through interviews and focus group discussions among stakeholders in two Mozambican health facilities and organizational readiness for implementing change (ORIC) assessments. In Aim 2, we will leverage data collected in food intake questionnaires administered under the parent study to assess the presence of longitudinal or dose-response relationships between exposure to community education modules and food intake. This formative research will contribute to scarce evidence for effective systems approaches to treat and prevent cardiometabolic disease in LMICs. This research plan will provide the F31 candidate rigorous predoctoral training including 1) application of implementation science frameworks for qualitative research, 2) development of participatory research methods, and 3) advanced statistical analyses using quantitative data for implementation science.