University Of California, San Diego

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT COVID-19 vaccines have been inequitably distributed and vaccine uptake has lagged, particularly for immigrant, refugee, Latino, and Black, Indigenous, People of Color (BIPOC) communities. Underlying reasons for the slow and variable uptake of COVID-19 vaccines include individual (e.g., health literacy, trust), cultural (e.g., linguistic needs), and structural barriers, such as technological and systemic factors (e.g., complex and onerous processes to schedule and attend vaccine appointments). There is no one-size-fits-all approach to vaccine implementation. This is borne out in the data indicating enormous disparities in COVID-19 testing, morbidity, mortality, and inoculation rates. Culturally relevant and linguistically appropriate, dynamic, and scalable strategies are essential to the immediate and long-term success of COVID-19 vaccine uptake and pandemic mitigation. Changing COVID-19 vaccine uptake behaviors offers an opportunity to concurrently improve engagement in other preventive health behaviors that are important public health priorities (e.g., diabetes management, cancer screenings, recommended adult vaccinations). We propose to co-refine, test, and scale a multicomponent health program to address the multi-level barriers to vaccine uptake and engagement in preventative healthcare in immigrant, refugee, Latino, and BIPOC communities in San Diego. Harnessing reverse innovation thinking, we will apply the Practical Robust Implementation and Sustainability Model (PRISM) to co-create with our community partners, the Global ARC, and San Ysidro Health, the elements of a health program that combines mHealth outreach (community-driven text and voice messages) and enhanced care coordination. Responsive to NOSI NOT-MD-22-006, this R01 will build on our current CEAL and RADx-UP implementation research to scale and sustain a multicomponent health program to increase acceptance, access, and uptake of COVID-19 vaccination and preventive care engagement among underserved communities. We assembled an experienced team of community-engaged implementation scientists, health equity, public health, and infectious disease researchers to accomplish the following aims: 1) Optimize a multicomponent health program to promote COVID-19 vaccine uptake and engagement in preventive healthcare using our established co-creation approach to address multi-level (individual, community, systemic) barriers to vaccine uptake and preventive care engagement; 2) Evaluate the implementation, effectiveness, and sustainment of the multicomponent COVID-19 vaccine and preventive care engagement program using a hybrid type 3 implementation-effectiveness sequential multiple assignment randomized trial design across immigrant, refugee, Latino, and BIPOC communities in Central and East San Diego.

NIH Research Projects · FY 2026 · 2022-09

PROJECT SUMMARY/ ABSTRACT Severe hypoglycemia remains a common and life-threatening issue for patients living with type 1 diabetes (T1D). Research has consistently shown that patients with impaired awareness of hypoglycemia (IAH), which typically coexists with a diminished counterregulatory response (CRR), are at the highest risk for severe hypoglycemia. However, we currently do not have clinically available tools to identify patients with IAH, and even if identified, we lack interventions to reduce their risk. The ultimate goal of this research is to address these large, unmet needs. Specifically, we plan to utilize continuous glucose monitoring (CGM) to identify which CGM metrics are associated with IAH and a diminished CRR (AIM 1). We will then determine if modern T1D management with hybrid closed loop (HCL) systems can restore hypoglycemia awareness (AIM 2). AIM 1: Identify the CGM metrics associated with hypoglycemia awareness Our first hypothesis is that time below range (TBR) by CGM will inversely correlate with epinephrine AUC during a hypoglycemic clamp. This hypothesis is strongly supported by our preliminary data detailed in our research strategy. To test this hypothesis, we propose that the consortium enroll a total of 112 subjects with T1D. Subjects will wear a blinded CGM for 10 days prior to a hypoglycemic clamp. Data to be collected will include counterregulatory hormones during hypoglycemia, hypoglycemia symptom scores (Edinburgh), and hypoglycemic awareness questionnaires (Clarke, Gold, Pederson, and hypo A-Q) as we have previously done. CGM metrics will be correlated with CRR, symptom scores, and currently used questionnaires to define awareness. The study design will determine which CGM metrics predict hypoglycemia awareness and CRR. AIM 2: Determine if hypoglycemia awareness can be restored in individuals with T1D using up-to-date management of diabetes Our second hypothesis is that reducing TBR using modern diabetes management with HCL systems will improve awareness of hypoglycemia and the CRR. To test this hypothesis, all 112 subjects will be randomized 1:1 to either a control arm or a hypoglycemia reduction arm for 2 years. In the control arm, all insulin delivery methods will be allowed and CGM targets will follow current standards of care that allow up to 4% TBR (~1 hour/day). The hypoglycemia avoidance arm will provide all patients with a HCL system and target <1% TBR. Hypoglycemic clamps will be conducted at baseline and months 3, 6, 12, 18, and 24 with 10 days of blinded CGM prior to each clamp. The primary outcome will be the difference in epinephrine AUC during hypoglycemic clamp between study arms. This study design will determine if up-to-date T1D management can restore the CRR and hypoglycemia awareness.

NIH Research Projects · FY 2025 · 2022-09

Project Summary The cellular environment is crowded with a concentration of macromolecules between 200-400 grams per liter. This crowding affects protein interactions, binding affinities, and diffusion. These conditions are not replicated in the dilute solutions used for in vitro studies. To have a full understanding of protein function, it is necessary to study proteins in complex environments that mimic the in vivo environment. However, the high concentration of macromolecules makes it difficult to perform structural studies in these systems. Owing to this, it is necessary to develop new methods to study protein structure in complex model systems. Here, we propose to further establish the protein footprinting method fast photochemical oxidation of proteins (FPOP) for studying complex model systems. FPOP utilizes hydroxyl radicals to oxidatively modify solvent accessible amino acids in proteins. The in vitro method can identify protein-ligand and protein-protein interaction sites as well as regions of protein conformation changes. My group has further expanded FPOP for studies in cells (IC-FPOP) and in vivo (IV-FPOP) in C. elegans, an animal model for human disease. We have demonstrated that IC- and IV-FPOP can oxidatively modify hundreds to thousands of proteins in these complex systems. The next step in method development is to establish their efficacy for identifying protein interactions in these model systems by studying specific applications. For the next 5 years, we plan to apply IC- and IV-FPOP to study protein folding and aggregation. The identification of protein interactions involved in misfolding and aggregation will help design new therapeutics. We also plan to extend the method into another three-dimensional model system, ex vivo tissue. This will provide structural information in a model system that more closely resembles the in vivo environment than monolayer cell culture.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY This proposal leverages cephalopods as a uniquely suited model system to ask how sensory systems detect and discriminate diverse environmental signals. Octopuses, as representative cephalopods, use their flexible arms and semi-autonomous distributed nervous system to explore their surroundings at a distance by locally detecting and capturing prey. This unique “taste by touch” system is mediated by chemotactile receptors (CRs), which are structurally similar to nicotinic acetylcholine receptors, but are insensitive to neurotransmitters, and instead detect poorly soluble molecules to mediate contact-dependent aquatic sensation. Here, we will exploit cephalopod chemotactile systems to understand how subtle evolutionary modifications in single proteins facilitate a functional transition from neuronal signaling to environmental sensation. Our approach spans structural biology to animal behavior. First, in Aim 1, we propose to determine high resolution structures of ligand- bound and apo octopus CRs to analyze structural and biophysical underpinnings of sensory versus neurotransmitter receptor function. In Aim 2, we will extend our comparative approach to CRs in distinct cephalopods with specific behaviors. In contrast to octopuses that use arms for active exploration, cuttlefish are ambush predators that strike and capture unsuspecting prey with their eight arms and two long tentacles. We recently discovered CRs in cuttlefish, which detect distinct ligands, exhibit different voltage dependence, and enable unique behaviors. Here, we will extend our analyses to include structurally informed experiments to compare aspects of ligand binding, ion permeation, and channel gating to ask how receptor function is suited to particular organismal behaviors. Finally, we recently found that octopus sensory cells express diverse CR subunit combinations that can form homo- and heteropentameric ion channel complexes. Subunit composition alters ligand sensitivity and ion permeation to tune signal detection, transduction, and filtering to influence peripheral processing in the octopus’ unusual distributed nervous system. In Aim 3, we will analyze the structural basis by which heteromeric complexes alter biophysical properties of ligand binding, ion permeation, and channel gating. Collectively, these studies will reveal broad principles underlying the structural basis for sensory receptor function and the evolution of biological novelty.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY COVID-19 cases and hospitalizations in children have increased dramatically worldwide. Although most COVID- 19 is mild in children, severe illness and post-infectious complications can occur. We and others have found that children are an important source of household and community transmission. Vaccination is the most effective way to prevent severe infection and decrease transmission. Infants under 6 months of age are at high risk for life-threatening complications, but a vaccine for this age group is not yet in clinical trials; thus, maternal vaccination and breastfeeding may be an important strategy to protect infects. SARS-CoV-2 infection and vaccine immunity studies have focused predominantly on adults, but children have developing immune systems and may respond to the new mRNA vaccination platform differently from adults. This proposal addresses the critical need to study the short- and long-term immune responses to COVID-19 mRNA vaccination in children, human milk, and breastfeeding infants. We have a successful ongoing longitudinal COVID-19 vaccination cohort that began in December 2020, in which we have collected biologic specimens from 368 individuals including adults, children, and lactating mother-infant pairs. We will enroll a total of 560 individuals down to 6 months of age after the mRNA vaccine receives Emergency Use Authorization (EUA) for the younger age group. Participants are followed every 3 months for nasal, saliva, milk (if lactating), and blood samples. We will test all COVID-19 symptomatic or exposed participants for breakthrough infection throughout the study period. Our central hypothesis is that the repertoire, magnitude, and longevity of COVID-19 vaccine- induced immune responses will be dependent on age and previous experience with SARS-CoV-2 infection. Importantly, our study will also move beyond the systemic immune responses to examine mucosal immunity in the respiratory tract and in human milk. To test the hypothesis, we will characterize vaccine induced serum, nasal, and saliva SARS-CoV-2-specific antibody response (Aim 1) and cellular (CD4+/CD8+) response (Aim 2) in children compared with adults and identify key immunologic correlates of protection against breakthrough infection. We will also determine humoral and cellular responses in human milk and secretory IgA in the breastfed infants’ upper respiratory tract and evaluate vaccine-induced differential gene expression in milk that direct the immune response (Aim 3). Our collaborative team with expertise in vaccinology, immunology, virology, epidemiology, and bioinformatics will ensure successful integrative analyses and interpretation of these immunologic and transcriptomic data. Completion of the study will provide a comprehensive characterization of longitudinal COVID-19 mRNA vaccination-induced immunity across age groups and in human milk to inform vaccination strategies to optimize the protection of children and infants.

NIH Research Projects · FY 2025 · 2022-09

The term blood-­brain barrier collectively describes the properties of central nervous system (CNS) vasculature  which stringently regulate the movement of ions, molecules and cells between the blood and neural tissue. These  properties protect the CNS from toxins, pathogens and CNS immune surveillance, and provide the neural tissue  the  necessary  nutrients  for  proper  function.  The  vascular  endothelial  cells  of  the  CNS  confer  many  of  the  properties  of  the  blood-­brain  barrier,  they  form  paracellular  and  transcellular  barriers,  and  possess  distinct  transport properties that efflux potential toxins and deliver nutrients. Blood-­brain barrier dysfunction is observed  in a wide array of neurological diseases including epilepsy, multiple sclerosis, stroke, Parkinson’s Disease and  Alzheimer’s  Disease.  The objective  of this  study  is  to  identify  novel  genes  that  allow the blood-­brain barrier to  regulate the health and function of the CNS. We identified several genes highly enriched in CNS endothelial cells  which are involved in the synthesis and degradation of monoamine neurotransmitters, dopamine and serotonin.  Dopamine  and  serotonin  are  neuromodulators  as  they  potentiate  the  firing  rate  of  neurons.  Outside  of  the  synapse they function as traditional extracellular signaling molecules and hormones. Neural circuits incorporating  dopamine  and  serotonin  are  involved  in  learning,  reward,  movement  and  mood,  while  dysfunction  in  monoaminergic  systems  are  observed  in  a  host  of  neurological  diseases  including  Parkinson’s  Disease,  Alzheimer’s Disease, neuropathic pain and neuropsychiatric disorders. Metabolism of dopamine and serotonin  by brain endothelial cells could be involved in the many processes and behaviors regulated by these monoamine  neurotransmitters.  To determine  the functions of blood-­brain barrier  monoamine neurotransmitter metabolism,  we generated conditional endothelial-­specific knockouts of Ddc, MaoA and MaoB and examined the requirement  of these genes in behavior and the brain levels of dopamine and serotonin. Our preliminary studies suggest that  blood-­brain barrier monoamine metabolism regulates the levels of monoamines in a behavior context-­dependent  manner. We will use the conditional endothelial-­specific knockout mouse models to examine how the blood-­brain  barrier  regulates  behavior  responses  to  sensory  stimuli  and  determine  the  mechanisms  by  which  blood-­brain  barrier  monoamine  metabolism  regulates  monoamine  neurotransmitter  levels  and  behavior.  Lastly,  we  will  examine  if  brain  monoamine  signaling  is  altered  in  endothelial-­specific  knockouts  of  blood-­brain  barrier  monoamine metabolic genes.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY FAIR DOs (Findable, Accessible, Interoperable, Reusable: Development Of Simulations) is a 5-year research education program in the modeling and simulation of digestive and renal neurobiology. It is developed by a team of award-winning faculty of educators and researchers who, among other things, is behind the FAIR mapping and knowledge management infrastructure for the SPARC effort. The main thrust of FAIR DOs is to educate the next generation of researchers and clinicians in the neural regulation of digestive and renal epithelia through taught modules and supervised modeling projects that make use of SPARC datasets, maps, and models. The FAIR DOs faculty will leverage teaching structures at Case Western Reserve University to recruit students. Overall, FAIR DOs aims to provide about 35/45 hours of training to over 75 students over the proposed five-year project, with each student research project producing one publication in the Physiome journal--launched by the International Union of Physiological Sciences. The SPARC Portal will provide findability to these student- generated open-access models and associated data via its Search and Flatmaps functions, as well as accessible interactivity via the O2S2PARC simulation platform. Overall, the FAIR DOs effort will provide a unique educational, mentored experience that will also generate a SPARC ecosystem of interoperating models that coherently study the neurobiology of digestive and renal epithelial transport. From a didactic “Systems” perspective, this ecosystem of models will be organized to follow the teaching narrative of the British Medical Association Award-winning “Medical Physiology 3rd Ed.” textbook, supplemented with recent literature, setting the stage to further leverage SPARC resources for education in the future.

NIH Research Projects · FY 2025 · 2022-09

Summary/Abstract The present equipment request is designed to support the NIH-funded BindingDB project. The equipment will make the BindingDB website and database faster and more reliable, so that researchers can make optimal use of BindingDB’s massive data collection. The purpose of BindingDB is described below. Medications are organic compounds that bind specific proteins in the human body, and researchers in universities, government labs, and pharmaceutical companies, are constantly at work seeking such compounds. These ongoing efforts generate a continuous flow of information about what small molecules bind what proteins, and how tightly. Scientists can learn from this information and use it to guide the discovery of the latest generation of medications. However, this information is typically published only in scientific articles or patents, where it cannot easily be found or accessed by other researchers. BindingDB makes these data findable and far more usable by extracting them from patents and articles and placing them in a web-accessible database equipped with a range of search, download, and analysis tools. For BindingDB to be maximally useful, this project also aims to maximize and, ultimately, certify, BindingDB’s responsiveness, reliability, and trustworthiness. The requested equipment will strongly support this aim.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Prenatal Alcohol Exposure (PAE) is a common cause of fetal growth restriction (FGR), which is a known risk factor for brain disability. Our previous studies identified extracellular maternal miRNAs as a causal link between PAE and FGR. These studies in pregnant women in Ukraine, resulted in discovery of 11 miRNAs (HEamiRNAs) that were elevated in plasma of heavy alcohol- exposed mothers who subsequently delivered growth-restricted, alcohol-affected infants (HEa), but not in exposed mothers who delivered infants that were apparently unaffected (HEua), or unexposed (UE) mothers. Maternal HEamiRNAs collectively explained 24-31% of the variance in infant height, weight and head circumference at birth, and in rodents, non-human primates, and in human trophoblast cell lines, explained PAE inhibition of placental trophoblast epithelial- mesenchymal transition (EMT) and FGR. Studies in this proposal, test an innovative hypothesis that two candidate cytokines, also identified in the Ukraine cohort, and extracellular miRNAs, control fetal growth in response to PAE, and that these may be manipulated to overcome FGR. In Aim 1, we test the hypothesis that two PAE-sensitive cytokines, C-reactive protein and sFlt1, control HEamiRNA transfer between maternal circulation and trophoblasts, as a means to inhibit the growth of chorionic villi, leading to FGR. Aim 2 is based on initial studies that showed 3 HEamiRNAs, which were elevated in both preeclampsia and FGR, promoted EMT gene expression in trophoblasts. We plan to test the hypotheses that cytokines, and sub-groups of birth outcome- defined HEamiRNAs may be manipulated to overcome FGR. Studies in Aims 1 and 2 will use cell culture and mouse models, with ultrasound imaging and transcriptomic studies, to assess the role of cytokines and HEamiRNAs in PAE-mediated inhibition of placental and fetal growth. In Aim 3, we will assess associations between HEamiRNAs, and other conditions linked to defects in placental development and function (preeclampsia, pre-term birth, spontaneous abortion, FGR), in samples from pregnant women recruited in the San Diego region with and without evidence of placental dysfunction, including PAE. We will additionally investigate the distribution HEamiRNAs in lipoprotein particles (LPPs) and extracellular vesicles (EVs) to determine whether pregnancy and/or placental dysfunction is associated with re-distribution of HEamiRNAs among extracellular compartments, possibly influencing their endocrine function and target tissue distribution. The proposed studies, led by qualified investigators with complementary skills, meet a significant need for mechanism-centered diagnoses and intervention for pregnancy complications due to PAE and other etiologies.

NIH Research Projects · FY 2024 · 2022-09

The term blood-­brain barrier collectively describes the properties of central nervous system (CNS) vasculature  which stringently regulate the movement of ions, molecules and cells between the blood and neural tissue. These  properties protect the CNS from toxins, pathogens and CNS immune surveillance, and provide the neural tissue  the  necessary  nutrients  for  proper  function.  The  vascular  endothelial  cells  of  the  CNS  confer  many  of  the  properties  of  the  blood-­brain  barrier,  they  form  paracellular  and  transcellular  barriers,  and  possess  distinct  transport properties that efflux potential toxins and deliver nutrients. Blood-­brain barrier dysfunction is observed  in a wide array of neurological diseases including epilepsy, multiple sclerosis, stroke, Parkinson’s Disease and  Alzheimer’s  Disease.  The objective  of this  study  is  to  identify  novel  genes  that  allow the blood-­brain barrier to  regulate the health and function of the CNS. We identified several genes highly enriched in CNS endothelial cells  which are involved in the synthesis and degradation of monoamine neurotransmitters, dopamine and serotonin.  Dopamine  and  serotonin  are  neuromodulators  as  they  potentiate  the  firing  rate  of  neurons.  Outside  of  the  synapse they function as traditional extracellular signaling molecules and hormones. Neural circuits incorporating  dopamine  and  serotonin  are  involved  in  learning,  reward,  movement  and  mood,  while  dysfunction  in  monoaminergic  systems  are  observed  in  a  host  of  neurological  diseases  including  Parkinson’s  Disease,  Alzheimer’s Disease, neuropathic pain and neuropsychiatric disorders. Metabolism of dopamine and serotonin  by brain endothelial cells could be involved in the many processes and behaviors regulated by these monoamine  neurotransmitters.  To determine  the functions of blood-­brain barrier  monoamine neurotransmitter metabolism,  we generated conditional endothelial-­specific knockouts of Ddc, MaoA and MaoB and examined the requirement  of these genes in behavior and the brain levels of dopamine and serotonin. Our preliminary studies suggest that  blood-­brain barrier monoamine metabolism regulates the levels of monoamines in a behavior context-­dependent  manner. We will use the conditional endothelial-­specific knockout mouse models to examine how the blood-­brain  barrier  regulates  behavior  responses  to  sensory  stimuli  and  determine  the  mechanisms  by  which  blood-­brain  barrier  monoamine  metabolism  regulates  monoamine  neurotransmitter  levels  and  behavior.  Lastly,  we  will  examine  if  brain  monoamine  signaling  is  altered  in  endothelial-­specific  knockouts  of  blood-­brain  barrier  monoamine metabolic genes.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Advances in synthetic biology provide powerful tools to interrogate the complex relationship between network structure and function. In this study, we will combine synthetic biology with computational modeling to investigate network-mediated regulation of cell damage and deterioration, a complex biological process. As similar studies in mammals are prohibitively time- and resource-intensive, we choose to focus on Saccharomyces cerevisiae, which has proven to be a genetically tractable model for many fundamental processes in mitotic cells and has allowed identification of many conserved genes that regulate cell-fate decisions in eukaryotes. Emerging questions include how these genes interact and how the interactions change dynamically to drive multi-generational cell deterioration dynamics. We recently found two distinct phenotypes in genetically identical yeast cells as they approach cell death: one with decreased ribosomal DNA (rDNA) silencing and nucleolar decline (Mode 1) whereas the other with heme depletion and mitochondrial decline (Mode 2). We found that stochasticity plays an important role in choosing one of the two paths, but once the fate decision is made, it is almost always irreversible. We identified a core molecular circuit, consisting of the lysine deacetylase Sir2 and the heme-activated protein (HAP) transcriptional complex, that governs the decision to select one of these two paths. Based on the model, we were able to engineer cells to follow a third path with a dramatically extended period of growth and survival, free of deterioration (Mode 3). In this proposal, we will expand these efforts and systematically perturb and rewire the core circuit that controls cell fate in order to reprogram its decision-making process. In Aim 1, we will use chemically-inducible promoters to control expression of Sir2 and HAP and thereby modulate cell-fate decisions in isogenic cells. We will use microfluidics to generate distinct, dynamic patterns of Sir2 and HAP expression and evaluate their effects on damage accumulation, physiological changes, and cellular decline. In Aim 2, we will genetically rewire the core Sir2-HAP circuit under the guidance of computational modeling and examine how these engineered circuits govern cell- fate decisions and cell deterioration dynamics. In Aim 3, we will use high-throughput microfluidics to identify the gene expression programs associated with Mode 1, Mode 2, and Mode 3 and examine how perturbations of these programs affect multi-generational deterioration dynamics. These analyses will uncover the genes and processes that underlie the missing connections between the Sir2-HAP core circuit and downstream modules that underlie cellular decline leading to cell death. They will enable us to expand our computational model and improve its predictive power. Throughout the study, we will construct deterministic and stochastic models, which will produce testable predictions and guide engineering of synthetic gene circuits. If successful, this research will advance a quantitative and predictive understanding of cellular fate decisions and cell deterioration.

NIH Research Projects · FY 2025 · 2022-09

PROJECT ABSTRACT The overarching goal of the “Partnership for Advancing Cervical Cancer Prevention in Women Living with HIV” is to improve the cascade of cervical cancer prevention for women living with HIV (WLWH) including increasing uptake of cervical cancer screening services, improving management of screen positive women, and optimizing precancer treatment outcomes, particularly in resource-constrained settings. Research base members have established collaborations with a history of conducting successful clinical trials and impacting clinical care. As a CASCADE Research Base, we will provide impactful scientific and statistical direction for pragmatic clinical trials to be conducted at the CASCADE Clinical Sites. We will establish a central administrative infrastructure to facilitate the conduct of three pragmatic trials. The proposed trials will include an investigation of primary HPV screening to facilitate uptake of cervical cancer screening, triage strategies for women who screen positive with high-risk HPV, and adjunctive treatment to improve outcomes to cervical pre-cancer treatments. We will also provide opportunities for early-stage investigators to develop clinical research skills within CASCADE. We have administrative expertise and experience in operationalizing international clinical trials and supporting clinical research sites in the timely implementation of projects. Our highly experienced investigators and our proposed trials will be of substantial benefit to the CASCADE network and the shared goal of eliminating cervical cancer in WLWH.

NIH Research Projects · FY 2026 · 2022-09

Project Summary The materno-fetal interface is a crucial environment for a healthy fetal development during pregnancy and delivery. A decrease in maternal trophic factors, such as maternal thyroid hormones or a maternal inflammation/maternal immune activation (MIA) have been linked to an increased risk for neurodevelopmental disorders, such as autism or preterm births. However, there has been no study to date to directly evaluate their impact on human developing brain. To fill this gap of knowledge, we propose to experimentally study the consequences of inflammatory factors on human neurodevelopment using the cerebral organoids co-cultured with microglial cells, a 3D model that mimics the early stages of the human brain development. Microglial cells are immune cells of the central nervous system that are generated in the yolk sac and penetrating the brain parenchyma during a critical time of the embryogenesis coinciding with synaptogenesis and gliogenesis. Given that microglial cells are generated in the periphery, we hypothesized that microglial cells are an important component of the human brain development and most likely the first brain cell type to be exposed and respond to an environmental factor/toxin that can potentially contribute to neurodevelopmental disorders prenatally. Here, we will investigate the impact of maternal inflammation due to bacterial or viral infections on human microglial cells and how that can impact the human developing brain. In addition, given that a person could be exposed to more toxic factors simultaneously or sequentially during that individual’s lifespan or during pregnancy, we will also aim to closely replicate the human experience, by testing the impact of co-exposures to infectious agents and study their synergistic impact on human microglial function and human neurodevelopment. Current studies using iPSCs has been limited sample size and the lack of diversity in the samples that are often derived from donors from a single race/ethnical group. Thus, to have a more inclusive approach, in this proposal, we will use a large cohort of samples from healthy individuals (>50), representative of different races, ethnicities and genders closely mimicking the proportions found in the United States’ population that will provide a better understanding of the health disparities and interventions both in populations and in specific subgroups. Ultimately, our goal is to provide a better understanding of the disease mechanisms involved in maternal inflammation and better targeted therapies from which the majority of the population in the US could benefit from.

NIH Research Projects · FY 2025 · 2022-09

Summary Alcohol use disorder (AUD) causes 1 in 20 deaths worldwide and imposes huge economic costs on society. Twin studies have shown that the risk for developing AUD is heritable. Genome wide association studies (GWAS) have indicated that, like most psychiatry diseases, AUD is highly polygenic. Although GWAS in both humans and rodents are powerful techniques, with different strengths and weaknesses, techniques to integrate the two are poorly developed. GWAS identify individual SNPs that influence a trait; because those SNPs are species specific, polygenic risk scores (PRS) and similar approaches cannot be used to transfer information across species. To address this limitation, we are proposing a framework for transferring polygenic signals across species. We introduce the concept of polygenic transcriptomic risk scores (PTRS). Whereas PRS sum the effects of many SNPs, a PTRS sums the effects of genetically predicted transcript abundance across many genes. Because these effects are at the gene, rather than SNP level, they can be applied to orthologous genes in other species. The extent to which a PTRS for AUD that was developed in humans might predict rodent behaviors believed to be relevant to AUD is currently unknown. In this grant we will assess whether PTRS can be used to translate polygenic signals related to AUD between humans and rodents. We focus on AUD because of the existence of high quality human GWAS data about AUD and related traits like alcohol consumption. In Aim 1, we will phenotype 1,250 HS rats for multiple alcohol self-administration traits. In Aim 2, we will perform GWAS and transcriptome wide association analysis (TWAS) for alcohol-related traits in the rats from Aim 1. In Aim 3, we will build PTRS for AUD and related traits and optimize them for portability across species. These aims address a critical limitation, namely the inability to transfer polygenic knowledge between species, which is inhibiting progress towards a deeper understanding of how polygenic liability for AUD alters molecular and cellular processes, brain circuits and behaviors. If successful, our results will open new avenues for research aimed at prediction, prevention, and treatment of AUD.

NIH Research Projects · FY 2025 · 2022-09

There is a great need to determine molecular signatures of Alzheimer’s disease (AD) to better understand AD pathophysiology, to detect AD in the preclinical stage, and to identify novel therapeutic targets. The plasma proteome is an ideal resource in which to identify molecular signatures, as proteins perform essential biological functions, are direct therapeutic targets, and shed light on disease mechanisms. Our preliminary data identified several plasma proteomic biomarkers associated with cognitive impairment and AD-related brain atrophy. We also found that accelerated biological (i.e., proteomic) aging relative to chronological age, as measured by our validated proteomic signature of aging (known as a ‘proteomic clock’), was associated with higher risk of multiple age-related conditions, including cognitive impairment. Yet, study of the proteomic changes preceding AD is still in its early stages. The objective of this study is to improve understanding of the proteomics of AD and related dementias (ADRD) by leveraging a nested case-cohort of 2,836 women in the racially diverse Women’s Health Initiative Memory Study (WHIMS). WHIMS contains longitudinal cognitive and neuroimaging measures; 1,336 incident cases of MCI and ADRD rigorously ascertained during 26 years of follow-up; rich phenotypic data; and preserved biospecimens. SOMAscan, the most comprehensive proteomics platform measuring 7,000 clinically relevant human proteins across numerous biological pathways, will be used to characterize the proteome longitudinally from blood samples collected at baseline and 14-18 years later. We will also obtain longitudinal plasma biomarkers of AD pathology from these samples. Our central hypotheses are that: (i) accelerated proteomic aging will be associated with higher risk of MCI/ADRD and lower likelihood of cognitively healthy longevity; and (ii) the 7,000-protein SOMAscan will enable identification of novel proteomic biomarkers, signatures, and biological pathways for MCI/ADRD and related endophenotypes. Our Aims are: Aim 1) Determine associations of validated proteomic clocks of aging with incident MCI/ADRD and cognitively healthy longevity (i.e., survival to age 90 without cognitive impairment); Aim 2) Determine associations of the plasma proteome at baseline, and 14-18-year changes in the proteome, with incident MCI/ADRD and cognitively healthy longevity; Aim 3) Relate proteomic clocks of aging and ADRD-associated proteins identified in Aim 2 to neuroimaging measures and plasma biomarkers of AD pathology; and Aim 4) Identify novel multi-protein signatures that predict MCI/ADRD, cognitively healthy longevity, and plasma biomarkers of AD pathology. Key proteins will be validated on a separate platform (ELISA) and replicated in external cohorts. This study will advance understanding of the heterogeneous mechanisms of ADRD pathogenesis and cognitive impairment in aging, identify risk and protective proteomic factors, and suggest candidate proteins for pathophysiology-targeted interventions in ADRD. The novel proteomic data will be a rich and valuable resource for the broader scientific community to identify novel proteomic biomarkers for a wealth of phenotypes, thus having an enduring impact.

NIH Research Projects · FY 2026 · 2022-09

Project Summary Prenatal alcohol exposure is estimated to impact 1-5% of children in the US alone. Yet, many individuals who have been exposed prenatally to alcohol and suffer from fetal alcohol spectrum disorders (FASD) fail to be recognized. This failure is due, in large part, to a paucity of specialized clinics and expert dysmorphologists who are trained to identify FASD. Access to appropriate diagnostic expertise is particularly limited in remote areas. Unfortunately, if individuals with FASD are not recognized and diagnosed, they do not receive critically needed services nor can they access potential interventions early in life, when intervention is most likely to be effective. The CIFASD5 Diagnostic-Telemedicine Resource (DTR) will ensure consistent and accurate assessment of the physical characteristics of FASD across the CIFASD research sites. In addition, the DTR will address the critical need for increased diagnostic capacity through training of non-expert practitioners across CIFASD5 sites using telemedicine-based methods. In the previous CIFASD4 iteration, telemedicine approaches were tested for this purpose in a small sample of clinicians and found to be an effective method for training and monitoring of new examiners. In CIFASD5, this method will be extended to multiple sites and be employed in the evaluation of over 1,800 children and adults. However, telemedicine alone is insufficient to expand capacity and ensure consistency and accuracy of diagnosis. To that end, several novel eHealth tools have been developed to assist in the detection of physical features associated with prenatal alcohol exposure. These tools hold promise in providing simple and efficient ways to screen and identify FASD. These include MorpheusQ, a smart-phone based app that automates facial feature detection; Face-to-Gene, a 2D facial image diagnostic aid used by clinical geneticists to screen for potential syndromes; and 3D facial image signatures. These tools are scalable and have the potential to improve screening and diagnosis across the globe, even in remote areas, such as in Alaska. However, the diagnostic accuracy of these tools needs to be systematically compared to standardized dysmorphological exams. It is essential to determine whether eHealth tools can effectively replace and/or improve traditional exams before they are widely implemented.

NIH Research Projects · FY 2025 · 2022-09

Abstract Our overall goal is to determine a potential causal link between L1, an active retrotransposon element, in human brain cells and Alzheimer's Disease (AD). L1s are involved in numerous human diseases with different molecular and cellular mechanisms. L1 expression was also found to be upregulated in AD postmortem human tissues, leading to the speculation that L1s could contribute, at least partially, to the mechanisms leading to AD in humans. However, despite all these correlations, there is no experimental-based causal link between L1s and AD so far. Here, we hypothesized that L1 expression in AD-derived brain cells can contribute to AD pathology, using both cell-autonomous and non-cell-autonomous mechanisms. Accumulation of L1-derived ssDNA molecules in the cytoplasm of glial cells might be responsible for a chronic low-level stimulation of the immune system, aka “sterile” inflammation, and can aggravate molecular and cellular phenotypes in neurons derived from AD patients. Moreover, a cell autonomous mechanism, triggered by L1 ssDNA accumulation in neurons could accelerate and exacerbate molecular and cellular phenotypes related to AD pathogenesis. Thus, the combinatory impact of L1 expression in the different cell types could causally contribute to AD. To experimentally test this hypothesis, we will use induced pluripotent stem cells (iPSC) from controls and AD to overexpress L1 ssDNA molecules and analyze their impact on different neuronal types individually or in combination using a brain organoid model system. The use of iPSC-derived cells and organoid model is perfectly suited as we can isolate the aging effect and capture the entire genome of AD individuals in relevant cell types. We have designed the following specific aims to test our hypothesis: Aim 1: Determine the impact of L1 retrotransposons in AD-derived astrocytes. We will measure alterations in transcriptomics, cytokines/interferon release levels and interferon-stimulated gene (ISGs) expression in AD/isogenic-derived astrocytes upon L1 ssDNA overexpression. We will also evaluate neurotoxicity using astrocyte conditioned media from AD-derived astrocytes overexpressing L1 ssDNA. Aim 2: Determine the impact of L1 retrotransposons in AD-derived cortical neurons. We will measure Tau aggregation and phosphorylation, and synaptic loss, all early hallmarks of the AD progression. Aim 3: To model AD progression with a brain organoid model. We will use our optimized protocol to generate brain cortical organoids (BCO) infused with human microglia from AD- and control individuals. BCO with L1 ssDNA but treated with nucleoside reverse transcriptase inhibitors (NRTI) as a protective agent, will be compared. Our proposal aims to demonstrate an eventual causal contribution of L1-derived ssDNA to molecular, cellular and network phenotypes in brain cells and organoids derived from AD individuals. Our data will reveal unexplored pathways with immediate therapeutic relevance that could lead to transformative treatments for AD and other aging-related syndromes.

NIH Research Projects · FY 2026 · 2022-09

ABSTRACT For decades, biologists have taken parts from disparate proteins and fused them in various combinations to create engineered variants with user defined properties. Despite the success of many of the generated tools (e.g. chimeric antigen receptors and enhanced CRISPR variants) the methods by which these proteins are discovered are slow and labor intensive, limiting our exploration to only a tiny fraction of potential protein space. Here, we introduce BArcoded Combinatorial Engineering and Screening (BaCES), a method that enables the simultaneous assembly and parallel testing of tens of thousands of combinatorial protein variants. The objective of this proposal is to use BaCES to create a new generation of enhanced Cas9-based transcriptional regulators, which will be combined with a novel experimental paradigm to probe gene function within in vivo contexts. The rationale underlying this proposal is that, if successful, we will create several transformative technologies and gain insight into the mechanism by which neurons tolerate neurodegenerative insults. Herein we demonstrate the feasibility of our BaCES platform and provide evidence supporting our unique approach to in vivo screening. To further our research goals, we will: 1) use BaCES to generate and quantify the behavior of 27,000 Cas9 activators and repressors; 2) thoroughly validate across targets and cell types a new generation of highly-potent Cas9 transcriptional modulators; and 3) apply these tools to perform a set of in vivo genetic screens to uncover regulators of neuronal survival within a mouse model of Parkinson’s Disease. This proposal is innovative from a technical perspective in that it creates a new method for rapidly searching through combinatorial protein space and implements a new paradigm for performing in vivo CRISPR screens within a complex cellular environment. It is also innovative in approach as it utilizes a high-throughput platform to gain insight into the genes and pathways that regulate neuronal survival within an in vivo model of disease. This work is significant in that it will create a novel method for performing combinatorial protein screens, identify a set of enhanced Cas9 activators and repressors to enable global research endeavors, and uncover the biological processes that neurons use to tolerate neurodegenerative disease- associated stressors. Our track record of producing widely adopted CRISPR tools, combined with our preliminary data demonstrating the feasibility of the proposed work and a group of long- standing committed collaborators, makes our team uniquely suited to carry out the outlined interdisciplinary research.

NIH Research Projects · FY 2025 · 2022-09

Summary This R01 renewal application is focused on triple negative breast cancer (TNBC), which is an aggressive life- threatening disease with poor prognosis and increased likelihood of recurrence and distant metastasis. Advances in cancer immunotherapy have demonstrated that modulation of the patient’s immune system can result in dramatic antitumor activity. The most promising immunotherapy approaches are those that are personalized and take advantage of the unique neoantigens within each patient’s tumor. Toward this goal, we developed a plant virus nanoparticle immunotherapy approach that activates innate immune cells within the tumor microenvironment (TME) to launch adaptive, systemic, and durable antitumor immunity. Specifically, intratumorally injected cowpea mosaic virus (CPMV) demonstrates potent efficacy in multiple mouse models, incl. TNBC. Trials in companion dogs with breast cancer also demonstrate potent antitumor efficacy. During the previous funding cycle, we gained insights into the mechanism of action and demonstrated that CPMV is recognized by pathogen-associated molecular pattern (PAMP) receptors that detect danger signals and activate the innate immune system; specifically, CPMV is recognized by Toll-like receptors (TLR2, 4 and 7). Further, we developed and tested combination and dual-pronged treatment approaches: we demonstrated efficacy of CPMV as solo-treatment as well as in combination with radiation, chemotherapy, immunomodulatory drugs, and checkpoint inhibitors, amongst others. This proposal builds on this strong portfolio of data. Our first goal is to focus on dual-pronged CPMV that combines its immunomodulatory and antitumor immunity properties with checkpoint therapy (Aim 1). Checkpoint blocking antibodies are effective at removing inhibitory signals but as monotherapy have variable and limited efficacy. In situ vaccination with CPMV increases tumor antigen specific effector T cells and our preliminary data indicate that CPMV treatment synergizes with immune checkpoint therapy. Next, we seek to develop targeted approaches that effectively concentrate systemically administered CPMV in tumors and provide further therapy options to treat metastatic disease (Aim 2). S100A9- targeted CPMV will be studied: expression of S100A9 (also known as myeloid-related protein 14 [MRP-14]), is linked to inflammation and carcinogenesis. Higher S100A9 expression in breast cancer correlates with a worse prognosis. S100A9 expression is an early event in tumorigenesis, enhancing tumor aggressiveness and metastasis. In the TME, S100A9 is secreted to the extracellular matrix, making it a highly suitable target for nanomedicine. Recognizing the potential of S100A9 as a pharmacologic target, we developed S100A9-targeted CPMV that efficiently concentrates at sites of metastasis enabling potent efficacy preventing outgrowth of metastases. Here we set out to detail the mechanisms of action and understand the pharmacology of S100A9- targeted CPMV. Finally, the preclinical development will be substantially extended with veterinary clinical trials in companion dogs with mammary tumors (Aim 3).

NIH Research Projects · FY 2024 · 2022-09

Project Summary / Abstract Chronic uncontrollable stress can precipitate or exacerbate many highly prevalent and debilitating neuropsychiatric disorders such as major depression and schizophrenia. Such stress-related disorders often share common motivational symptoms that result in reduced engagement in activities in pursuit of once- desired outcomes. Dopamine plays critical roles in voluntary movement, motivation, and reward-based learning, but its precise contribution to self-initiated goal-directed behavior remains poorly understood. The dorsomedial striatum (DMS) is well established in supporting goal-directed behavior and receives prominent dopaminergic input from the substantia nigra pars compacta (SNc). Anatomical inputs to these nigrostriatal dopamine neurons have been identified, but little is known about how this circuitry regulates nigrostriatal dopamine dynamics during goal-directed action. Furthermore, chronic stress manipulations in rodent models have revealed complex effects of stress on the adjacent mesolimbic dopamine projections to the ventral striatum, as well as structural and physiological alterations of corticostriatal inputs to the DMS. However, the effects of stress on nigrostriatal dopamine and the circuitry regulating it during goal-directed behavior has not been well characterized. The proposed experiments therefore will address these critical gaps by examining nigrostriatal dopamine transmission (Aim 1) and the striatonigral circuitry regulating these dopamine dynamics (Aim 2) in mice performing goal-directed behavior. These K99 mentored phase experiments will entail the integration of modern optogenetic techniques with the candidate's expertise in recording dopamine using fast-scan cyclic voltammetry, and they will provide opportunities for acquiring advanced technical training with in vivo electrophysiology and cutting-edge viral circuit-manipulation techniques under the guidance of Dr. Xin Jin (mentor) and Dr. Ed Callaway (co-mentor). Training in this suite of systems neuroscience tools will permit subsequent R00 independent phase investigations of how chronic stress alters the functional circuitry regulating nigrostriatal dopamine during goal-directed actions and more complex cost-benefit decision making (Aim 3). These experiments will entail distinct stress manipulations implemented following further guidance from Dr. Byungkook Lim (consultant) and a novel decision-making task adapted from the candidate's doctoral work examining decisions involving tradeoffs between reward and effort. Collectively, the research proposed in this Pathway to Independence award will yield unprecedented insight into how chronic stress affects the circuitry regulating an under-examined dopamine pathway in goal-directed behavior and action selection; it will provide the technical training and career development to launch the candidate's independent research program; and it will reveal important additional questions for future investigations of mechanisms supporting motivated behavior and mental health.

NIH Research Projects · FY 2025 · 2022-09

Project Summary. Human ALIX (also known as PDCD6IP) functions in endo-lysosomal pathway, apoptosis, enveloped virus budding, and other essential cell signaling and membrane scission processes. These diverse functions are regulated by its posttranslational modifications (PTMs), specifically tyrosine phosphorylation and ubiquitination. We recently uncovered that ALIX, through its proline-rich domain (PRD), forms liquid-like condensates and amyloid fibrils, and that both these assemblies dissolve on phosphorylation and reform on dephosphorylation of its tyrosine residues. Projects in this ESI-MIRA proposal expand upon these exciting discoveries and will uncover the dynamic functional interplay between phase separation, fibrillization, and PTMs of ALIX. Specifically, we will: determine the structural characteristics of ALIX’s assemblies, their regulation by tyrosine de/phosphorylation and membranes, and their formation in mammalian cells (direction 1), elucidate the functional relevance of ALIX polymerization, and the mechanisms of the time-dependent transitions of ALIX condensates to fibrils (direction 2), and characterize the interactions between ALIX and ubiquitin, and determine the cross-talk between ALIX ubiquitination and its phosphorylation-mediated polymerization (direction 3). Structural characterization of ALIX’s higher- order assemblies in direction 1.1 will reveal the interactions hotspots that govern its phase separation and novel atomic- resolution details of how a PRD can form β-sheet rich fibrils. Mechanistic studies in direction 1.2 will elucidate regulation and modulation of ALIX condensates and fibrils by lipid membranes and tyrosine de/phosphorylation, revealing how a kinase accesses its sites within these assemblies, and the identity of tyrosine residues whose dephosphorylation triggers ALIX polymerization. Cellular studies in direction 1.3 will examine ALIX polymerization in mammalian cells. In direction 2.1, we will determine how polymerization affects ALIX’s functions. Mechanistic studies in direction 2.2 will elucidate time-dependent hardening of ALIX condensates into fibrils, yielding new insights into the role of phase separation in fibrillization. Structural and kinetic studies in direction 3.1 will elucidate the interactions between ALIX and ubiquitin. Finally, in direction 3.2, we will determine the impact of ALIX ubiquitination on its phosphorylation-mediated polymerization. The above studies build upon our discoveries of the unique ALIX assemblies, their modulation by PTMs, the slow maturation of ALIX condensates into rigid fibrils, residue-specific details of ALIX – late endosomal membrane interactions, and how ALIX’s phosphorylation inhibits these interactions. Extensive preliminary results, including highly homogenous samples of ALIX assemblies enabling their structural characterization, the discoveries of selective recruitment of ALIX’s signaling partners in its condensates, and of ALIX – ubiquitin interactions in solution, assure high feasibility of successfully completing our proposed studies. Our newly developed methods, including a new labeling strategy to facilitate NMR studies of ALIX assemblies, and the production of milligram quantities of pure ubiquitinated proteins, promise groundbreaking insights into ALIX polymerization and the role of ubiquitin in ALIX biology. Collectively, these studies will define molecular mechanisms that underlie ALIX’s multifaceted cellular and membrane functions.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Social isolation and loneliness are associated with increased risk for cognitive decline and Alzheimer’s disease (AD) in older adults. This is a pressing public health concern given worldwide increases in social disconnectedness. Yet, research on the effect of social disconnection, especially social isolation, on risk for AD is hindered by reliance on retrospective self-report measures of social relationships and behaviors. Moreover, potentially modifiable social cognition mechanisms (e.g., apathy, defeatist social appraisals, biased threat perception) that may differentially contribute to isolation and loneliness are poorly understood. Integrated digital technology measurement approaches using ecological momentary assessment (EMA), which involves multiple daily smartphone surveys about social behavior and experiences, and passive social sensing, including GPS location and quantification of social interactions using smartphone sensors, could provide more precise and reliable probes for detection of social disconnection related to risk for AD in CN older adults, and could also reveal novel modifiable social cognition treatment targets to mitigate risk. Measurement problems, such as incomplete and inconsistent coverage of daily social behavior and experiences, have hampered observational and interventional research. Our inter-disciplinary research group has led development and validation of EMA, mobile social cognitive testing, and scalable passive sensing (GPS and voice sensing) measures, and social network analyses, to more precisely quantify social dynamics in daily life. We have also translated our real-time EMA data into interventions that reduce social cognitive biases that influence day-to-day social disconnection (e.g., social threats, defeatist attitudes). For the first time integrating these tools, we propose to investigate associations between real-time maladaptive social cognitive biases, social isolation, loneliness and AD risk biomarkers in 128 cognitively normal (CN) older adults divided into high (N=64) and low (N=64) risk based on CSF P-tau181, A42 and subtle cognitive decline (SCD) markers. We propose to administer in-lab standard measures, as well as EMA, GPS and social interaction digital detection measures, of social isolation, loneliness and social cognitive biases. We propose to compare high- and low-risk CN groups on EMA (primary outcome), passive sensing and in-lab measures, and will also examine relationships between digital social relationship measures, in-lab measures, and biomarkers. The goals of the project are to show that EMA and passive social sensing measures (1) can differentiate high- and low-risk CN groups; (2) are associated with known Aβ and P- tau biomarkers; and (3) are associated with social cognition biases that can be modified using treatments like in-person and digital cognitive-behavioral therapy. The immense data and digital products of this study would be available for future research probing real-world social processes in older adults.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract The long-term goal of this research program is to identify negative consequences associated with e-cigarettes and other newer tobacco products, and reduce the health burden of tobacco use by developing interventions and identifying policies to reduce youth use. This proposal focuses on e-cigarette use, which has increased substantially and is now the most common nicotine product used by US youth. Our collective knowledge is not yet sufficient to identify factors that can be targeted in e-cigarette interventions, or to be sure that we understand the negative consequences of use. We propose to take advantage of the unique nature of the ABCD study, which includes regular assessments of a large, nationally representative cohort from ages 9 to 19. The design of the study will allow us to examine a range of both (1) modifiable risk factors for e-cigarette use and (2) behavioral, psychological, neurocognitive, and physiological consequences of use. The goals of this study are to prospectively test the following hypotheses: (1) e-cigarette initiation and progression in children is driven by modifiable social, contextual, and individual risk factors; (2) children who try e-cigarettes will be more likely to become users of combustible tobacco products; (3) youth e-cigarette use is associated with neural alterations and neurocognitive deficits; (4) e-cigarette use in adolescence is associated with negative health consequences. Our goals are twofold. First, we seek to identify modifiable factors among children that are associated with risk of chronic tobacco use and dependence, in order to inform intervention development. Second, we aim to delineate the extent to which e-cigarette use causes negative physical, behavioral and cognitive consequences, in order to inform policymakers who have regulatory authority over tobacco products. The overarching goal is to reduce tobacco use among youth and thus associated morbidity and mortality.

NIH Research Projects · FY 2026 · 2022-09

Modified Project Summary/Abstract Section A diverse biomedical workforce is critical for scientific innovation and health equity. Yet, underrepresented racial/ethnic (UR) scientists and women remain disproportionately low, particularly as faculty at academic research institutions. Multiple evidence-based strategies need to be implemented to enhance faculty diversity, inclusion and to create cultures of inclusive excellence. However, a knowledge gap exists regarding integrated strategies to address diversity and inclusion, the impact of faculty cluster hiring, and institutional change models on fostering inclusive excellence. The overall goal of the University of California San Diego (UCSD) NIH Faculty Institutional Recruitment for Sustainable Transformation (FIRST) Program is to: 1) foster sustainable institutional culture change, 2) promote institutional excellence by hiring a diverse cohort of faculty and 3) support faculty development, mentoring, sponsorship and promotion. The objectives are to: 1) demonstrate institutional support, develop or modify a strategic plan with elements that will be implemented to achieve systemic and sustainable institutional culture changes towards inclusive excellence, 2) develop an evaluation plan to assess impact on the institution towards achieving FIRST program goals, 3) conduct recruitment of new faculty, outline institutional commitments, and develop recruitment committees based on commitments to diversity, equity and inclusion, 4) establish a retention plan to secure institutional commitment and a supportive environment for new faculty hires, 5) establish individual research and career development plans, mentorship plans and 6) develop strategies to reduce isolation, increase community building and foster career development for all new faculty hires. The primary hypothesis is that a cohort model of faculty hiring, sponsorship, continual mentorship and support for professional development, embedded within an institution implementing evidenced-based practices to create academic cultures of inclusive excellence will achieve significant improvements in metrics of institutional culture and scientific workforce diversity. We propose the following Specific Aims. Aim 1. The aim of the Administrative Core is to provide strategic leadership, management, and administrative oversight to support the recruitment and retention UCSD FIRST faculty and oversee implementation professional development and institutional transformation for inclusive excellence. Aim 2. The aim of the Faculty Development Core is to use evidence based strategies to enhance FIRST faculty academic advancement, research success, and inclusive institutional excellence. Aim 3. The aim of the Evaluation Core is to develop and implement an evaluation plan to assess the institutional impact of the FIRST Program and to assist the Coordination and Evaluation Center evaluation of the combined FIRST Cohort Programs. Together, these aims will promote institutional culture change at UC San Diego towards inclusive excellence by using evidence-based strategies to enhance UR faculty recruitment, academic advancement, research and career development, integration and implementation of system-wide structured faculty development programs to enhance inclusivity.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract ABSTRACT Mitochondria not only provide 90% of the energy required for reading these very lines but they are also responsible for the correct differentiation of the cells lining your gut. The recent recognition that mitochondria play an active role in stem cell fate decisions has moved them from passive power plants to active centers of cell signaling. Our inability to automatically track mitochondria and link them to the fate of a cell is in stark contrast to the increasing relevance of this organelle in stem cell fate decisions and intestinal diseases such as Crohn’s disease and colorectal cancer. Mitochondria have always been too small and too fast for volumetric imaging and tracking. Our preliminary data show that a combination of recently developed lattice light-sheet microscopy and our in-house developed computational image processing pipeline can succeed in the four-dimensional tracking of the entire cellular mitochondrial network. Here we propose to 1) expand our prototype into a general tool that can track mitochondria in multiple cell types and network morphologies, to 2) elucidate the coupling between mitochondrial network morphology and cellular fate, and to 3) create a predictive model that links the mitochondrial network morphology to its underlying signaling drivers and to the fate decisions that are caused by different morphologies. We propose to use intestinal epithelial organoids and the differentiation of intestinal stem cells to paneth cells as a model system for a mitochondria-directed fate determination. This proposal will open a new window into mitochondrial biology that will translate to a large number of mitochondria-related diseases such as cancer and neurological disorders such as epilepsy, Parkinson’s disease, and Alzheimer’s disease.