University Of California At Davis

NIH Research Projects · FY 2025 · 2024-09

This application is a resubmission in response to the Funding Opportunity Announcement (FOA) identified as PAR-21-271. PROJECT SUMMARY In the US, cancer disproportionately affects many racial/ethnic minority communities, defining significant cancer health disparities (CHDs). Across all racial/ethnic groups, American Indians and Alaskan Natives (AIANs) experience one of the most substantial disparities with the highest comorbidity rates and lowest 5-year survival rates for all major cancer types. CHDs are driven by a combination of biological and social determinants of health, but actionable knowledge of both elements remains severely limited for AIANs. In this K99 proposal, I will address two critical problems that have perpetuated CHDs among American Indians/Alaskan Natives (AIANs): underrepresentation in precision medicine that limits discovery translation and race-level trends for outcomes and SDOH that conceal community diversity critical to effective implementation. Neither issue can be successfully addressed without the involvement and voices of Native communities, who have long been subjected to extractive research practices with little to no return on community health. This proposal centers Native communities living within the UCD Comprehensive Cancer Center catchment area (UCDCCC-CA) in a community-based participatory research (CBPR) framework to develop guidelines for appropriate Native community engagement for biomedical research while assessing community-level biological and social determinants of cancer health and care access. In Aim 1, I will utilize whole exome sequencing data from 300 Hispanic/Latino (HL) gastric cancer samples to identify somatic alterations that are associated with Indigenous American (IA) genetic ancestry at both global and local ancestry levels. These may be applicable to AIANs who also share high IA ancestry. Associated loci will be functionally validated in HL patient-derived organoid models in the R00 phase. In Aim 2, I will administer a validated survey and conducted focus groups interviews to identify key factors that contribute to individual decision-making for participation and biospecimen donation in cancer research among AIANs. In the R00 phase, I will work with UCDCCC-CA Native community stakeholders and my Native community advisory board to translate these findings into Native community engagement guidelines for research. In Aim 3, I will utilize cancer risk and health behavior data from the Aim 2 survey and cancer outcomes and sociodemographic data from the California Cancer Registry to elucidate community-level social determinant of health that drive UCDCCC-CA Native community cancer outcomes. In the R00 phase, I will use these findings to identify areas of intervention in collaboration with community and Native-focused health clinic partners to improve Native community cancer health. This proposal fulfills the mission of the National Institute for Minority Health and Health Disparities to understand and reduce health disparities. The results of this proposal will increase understanding of biological vulnerabilities and social determinants of health that lead to greater cancer incidence and mortality among AIANs, while creating the infrastructure to ethically engage Native communities and increase their participation in cancer research.

NIH Research Projects · FY 2024 · 2024-09

NOTE: You must submit in Word format, not PDF, for eRA to update all the systems. Metabolic health issues such as metabolic dysfunction-associated steatohepatitis (MASH) and type 2 diabetes melilotus (T2DM) are risks for hepatocellular carcinoma (HCC) and colorectal cancer (CRC). Emerging evidence has revealed that diet-induced gut dysbiosis contributes to metabolic disease development, including cancer. Thus, gut dysbiosis-targeted drug discovery should be explored, which is anticipated to advance the field of metabolic disease prevention and treatment. This project aims to develop and characterize an orally delivered nanodrug designed and synthesized to compensate for the shortage of specific microbial metabolite-regulated signaling in liver and colon cancer patients. The metabolic benefits of this novel drug have been uncovered in diet-induced obese (DIO) mouse models. Our data shows that human CRC and HCC specimens have elevated protein deacetylases and reduced signaling for short-chain fatty acids (SCFA), harboring histone deacetylase (HDAC) inhibitory properties. Through epigenetic mechanisms, HDAC inhibitors enhance intestinal retinoic acid (RA) signaling, which is consistently reduced in human HCC and CRC. We have validated the interactive and combined benefits of HDAC inhibitors and RA in cancer cells and animal models. We, therefore, have synthesized nano- “RA-based HDAC inhibitors” using polymers. One of the synthesized drugs, PRORA, was produced by covalently linking propionic acid (PRO), an HDAC inhibitor, and all-trans RA (RA) to polyvinyl alcohol (PVA), assembled into nano-micelles. This nano-formulation allows the co-delivery of both chemicals simultaneously. PRORA can be absorbed by the intestinal epithelial cells. In DIO mice, oral therapy with PRORA reverses diet-induced fatty liver, splenomegaly, and insulin resistance without noticeable toxicity. Based on this exciting success, we propose to uncover the mechanisms of action. Our data revealed that PRORA activated hepatic and intestinal FXR, a bile acid (BA) receptor, which also has reduced expression in human HCC and CRC. We propose analyzing the effects of PRORA in FXR-dependent or -independent manners using DIO wild-type mice and FXR KO mice, followed by transcriptomic profiling using colon and liver RNA. PRORA-influenced genes and pathways will be uncovered in the gut-liver axis. In addition, because FXR KO mice are prone to developing MASH and progressing into liver cancer, the proposed approaches allow us to determine whether PRORA has liver cancer-preventive effects. The data generated impact on having a safe orally delivered drug to treat or prevent MASH and T2DM.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY In all animals with internal fertilization, hundreds of seminal fluid proteins (SFPs) are transferred to females during mating. These SFPs have canonically been primarily thought of as a set of male proteins that advance male interests during post-mating processes, including by manipulatively influencing female behavior and reproductive investment. From this perspective, SFPs are thought to be an important substrate of interlocus sexual conflict against which females must continually evolve defensive countermeasures, which has been hypothesized to explain observations that SFPs evolve rapidly. Using single-nucleus RNAseq of the Drosophila melanogaster female reproductive tract (uterus, oviduct, glands, and sperm storage organs), I discovered that females express >40% of the 292 known D. melanogaster SFPs. Moreover, multiple female reproductive cell types specialize in producing SFPs prior to mating. For example, in a cell type in the seminal receptacle, SFP genes comprise 30% of mRNAs in the average sequenced nucleus (up to 65% of mRNAs in some sequenced nuclei). This result demands a re-evaluation of SFP biology that integrates the function and possible selection on the female-contributed protein fraction, which I take on in this proposal by asking: 1) What are the functions of female-expressed SFPs, i.e. shared reproductive secreted proteins? Do the female- and male-contributed protein fractions have the same function? And 2) How do shared secreted reproductive proteins evolve? Do the sex-shared secreted proteins exhibit different evolutionary modalities than the sex-limited SFPs? Do evolutionary and functional patterns in these genes better support a model of cooperation or sexual antagonism? Importantly, for the sex-shared proteins, any potential sexual conflict would have to be intra-locus conflict, with implications for the predicted duration and outcome of the conflict. I address these knowledge gaps using cutting-edge genome engineering methods, specifically gene knock-out and scarless CRISPR insertion of transgenic nanobody-epitope tagged proteins, to allow both immunofluorescent visualization and functional characterization. A second set of experiments uses comparative genomics and transcriptomics in a phylogenetic framework to determine the history of the proteins, the persistence of their expression by each sex, their evolutionary rate, and the evidence for either directional or stabilizing selection on the protein sequences. Because the cell types in D. melanogaster females that specialize in SFP production belong to the sperm storage organs, the proposed work will additionally test whether convergently evolved gains of sperm storage in a third female organ, observed among select non-melanogaster Drosophila species, are associated with gains of SFP expression in additional cell types. In total, the proposed work will provide a missing perspective on a famous set of genes in a premier genetic model organism. Simultaneously, the proposal provides critical training opportunities in future-oriented methodologies, toward achieving career independence.

NIH Research Projects · FY 2025 · 2024-09

SUMMARY/ABSTRACT Breast cancer is a leading cause of death in the U.S. and a major public health burden as one woman out of eight will receive a diagnosis in her lifetime. Advances in DNA genotyping and sequencing technologies have led to the identification of >300 germline variants that contribute to breast cancer risk. However, as of today, large ‘omics’ studies include women of mostly European ancestry even though preliminary evidence indicates that there is a clear benefit in studying genetically diverse samples. A genome wide association study (GWAS) in Hispanic/Latina (H/L) women with one tenth of the number of individuals included in previous studies led to the discovery of protective variants only present in populations that have Indigenous American ancestry. These variants significantly add to the predictive accuracy of breast cancer polygenic risk scores (PRS). H/L represent 16% of the U.S. population and in some states, up to 37%. To address the lack of H/L representation in genomics studies, we have created the Latin America genomics of breast cancer (LAGENO-BC) consortium, which includes ~28,500 Latin American breast cancer patients and ~32,600 unaffected women from 11 different countries and the U.S. diaspora. Our goal is to study the Latin American-specific genetic architecture of breast cancer and develop predictive models that account for variation by ancestry and geography. By combining existing studies, LAGENO-BC quadruples the prior H/L BC GWAS sample size and consequently, provides the power required for the discovery of additional variants contributing to breast cancer risk and for improving the predictive power of PRS for risk stratification in those populations. In the first aim, we propose to discover susceptibility loci and genes associated with breast cancer in H/Ls by conducting GWAS and ancestry-matched multi-tissue model-based transcriptome wide association studies with readily available RNA sequence data (TWASs). These analyses will be repeated for overall risk and subtype-specific risk. In the second aim, we will use known and new genetic risk variants to improve breast cancer risk prediction using ancestry-aware approaches and integrating existing models based on non-genetic risk factors. This work is expected to result in the discovery of new risk variants that are especially relevant to populations with an admixed Indigenous American ancestral background. In addition, utilizing ancestry-matched and multi-tissue models for TWAS, new associations linked to ancestry- specific and tissue-specific eQTLs are expected to be uncovered. It is also expected that PRS scores that incorporate H/Ls variants and assess regional and ancestry heterogeneity will be more predictive than a one-size-fits-all approach. Beyond the proposed work in this application, LAGENO-BC is building up a resource that will serve as a foundation for future collaborative studies addressing a broad range of scientific questions related to breast cancer genetics in H/L populations.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Food insecurity is a strong mediator in the relationship between poverty and breast cancer mortality in the United States. National data has identified adult women experiencing food insecurity have 54% lower odds of reporting breast cancer screening in the past two years when compared with food secure women. Counties with very high levels of food insecurity are significantly associated with later-stage breast cancer among women ages 65 and older. Early detection reduces breast cancer mortality rates and increases the likelihood of better treatment options. However, higher food insecurity has been associated with delayed utilization of, and reduced adherence to, recommended medical care among individuals and communities. Partnering with existing community programs that offer supplemental food assistance may provide an opportunity to encourage breast cancer screening in this harder-to-reach population. The proposed study will apply community-engaged research methods to improve access to breast cancer screening among a marginalized population that is experiencing food insecurity. The central hypothesis is that addressing barriers to breast cancer screening promotion for diverse communities of women with food insecurity can increase screening uptake, ultimately improving cancer health equity. We will test our hypothesis via the following Specific Aims: (1) Identify barriers and facilitators to implementing a cancer prevention intervention in a food distribution setting; (2) Develop a tailored community-based cancer prevention intervention to promote breast cancer screening among women with food insecurity; and (3) Evaluate the acceptability, appropriateness, and feasibility of the community-based intervention promoting breast cancer screening among women with food insecurity. To achieve Aim 1, I will collect qualitative data about food bank distribution events and the clients served using participant observation with semi-structured fieldnotes and interviews (n=20) and directed content analysis guided by the Consolidated Framework for Implementation Research to gain an in-depth understanding of factors and elucidate mechanisms that impact implementation. For Aim 2, I will use Intervention Mapping as a systematic process for planning a community-based intervention to promote breast cancer screening among diverse women who are served by the food bank. For Aim 3, I will conduct a pilot study of the intervention with the food bank, then survey women ages 40-74 years (n=50) to evaluate select implementation outcomes. This research stands to significantly improve the understanding of food insecurity as a social determinant of cancer health and provide an implementation model to promote cancer prevention with this at-risk population. The training plan will be facilitated by the mentorship of my sponsor and the exceptional facilities at UC Davis. This proposal describes an integrative and comprehensive training plan to support scientific and professional development and propel me toward my long-term goal of becoming an independent public health researcher.

NIH Research Projects · FY 2025 · 2024-09

There are 21 different connexin isoforms in human that play crucial roles in intercellular communication during development and physiology. At molecular level, six connexin proteins assemble as hexamers to form an hemichannel that is inserted in the plasma membrane. The docking of two hemichannels from adjacent cells form a gap junction channel (GJC); both hemichannels and GJCs are permeable to ions and small metabolites (<1 kDa). Several human connexin mutations and multiple pathological conditions exacerbate the activity of undocked hemichannel causing cellular dysfunction. A major limitation in the field is the lack of adequate pharmacological and genetic tools to dissect out the role of undocked connexin hemichannels versus their functions as GJCs. The objective of this proposal is to create recombinant nanobodies targeting Cx43 protein that differentially recognize hemichannels from GJCs to overcome this technical barrier. These nanobodies have reduced size (15 kDa) allowing higher tissue penetration, reduced immunogenicity and easy genetic manipulation and engineering. To achieve this goal, we have selected peptides corresponding to extracellular regions, N-terminal helix and intracellular loop of Cx43 protomer for llama immunization. To identify and generate recombinant Cx43 nanobodies present in this serum, we will use the immunized llama’s B-cells as source of nanobodies-coding RNA to produce a nanobodies-phage display library. The selected nanobodies will be screening to recognize Cx43 via ELISA, Western blot and immunofluorescence. In addition, we will assess the nanobodies capabilities to block Cx43 hemichannel activity in vitro and their wide applicability using heterologous expression system and disease models. Considering the compelling evidence for a pathological role of Cx43 hemichannels in various diseases and our willingness to share the resulting antibodies with the whole scientific community, the present project is highly innovative and significant.

NIH Research Projects · FY 2025 · 2024-09

Few studies have characterized disease ecology questions for pathogens with robust environmental stages that cross ecosystem boundaries. In recent decades, terrestrially derived protozoan infections have been increasingly reported in marine mammals. Toxoplasma gondii and Sarcocystis neurona are common pathogens in southern sea otters, but their definitive hosts are terrestrial (felids and opossums, respectively). En route from their terrestrial definitive hosts to a marine animal, parasite stages are subject to diverse environmental forces that determine whether they are effectively mobilized and sufficiently viable to infect a marine host. Intriguingly, we found that the diversity of protozoa genotypes in sea otters, including virulent strains, does not reflect parasite diversity in terrestrial hosts. We hypothesize that (i) environmental forces drive selection of virulent protozoan strains in marine ecosystems; and (ii) virulent parasite strains accumulate and persist in submarine vegetated habitats. Our objectives will generate novel and diverse datasets for an integrative Bayesian modeling approach to test how land-sea environmental forces shape the distribution, population structure and selection of virulent parasites in coastal ecosystems. Objectives are designed to answer two questions: Q1: How do environmental forces across land-sea habitats affect the genetic distribution, transport and survival of T. gondii and S. neurona in the nearshore? And Q2: Are submarine vegetated habitats (kelp and seagrass) hot spots for transmission of virulent pathogens for sea otters? By integrating field, genomics, stable isotope analysis and modeling approaches across land-sea boundaries, this research will provide a new framework to understand how habitat and climate dynamics shape the ecology of virulent pathogens in marine environments. Focusing on kelp and seagrasses has the potential to yield transformative insight on infectious disease transmission in the coastal ocean, with broad implications for diverse host-pathogen systems. Our fully mechanistic model includes numerous ecological processes and incorporates the previously unrecognized importance of submarine vegetation in concentrating pathogens and mediating predator-prey interactions that determine marine host infection patterns. Results will thus represent a fundamental advance in our current understanding of infectious diseases that cross coastal boundaries.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Multiple sclerosis (MS) is an autoimmune disease that gives rise to chronic neurological deficits. It is caused by an immunological attack on the myelin sheath leading to demyelination and axon degeneration. MS is the most common demyelinating disease of the CNS in young adults. 8,000-10,000 children and approximately 1 million adults are currently diagnosed with MS in the US. Furthermore, most MS cases result in frequent relapses. Currently available treatments can diminish the incidence of acute flare-ups, but do not prevent progressive axonal degeneration and neurological disability. Mesenchymal stem/stromal cells (MSCs) and MSC-derived extracellular vesicles (EVs) have been suggested as a promising approach for the treatment of MS, since MSCs and MCS-EVs can confer multifactorial therapeutic functions such as immunomodulation, neuroprotection, and angiogenesis. Our lab has established methods to isolate and expand early gestational placenta-derived MSCs (PMSCs) and demonstrated that compared with adult tissue-derived MSCs, PMSCs and PMSC-EVs possess superior immunomodulatory, neuroprotective and pro-angiogenic functions. In a preliminary study, we confirmed PMSC-EVs promoted re-myelination through the differentiation of oligodendrocyte precursor cells (OPCs) into mature oligodendrocytes and improved motor function outcomes in an experimental autoimmune Encephalomyelitis (EAE) mouse model of MS. One major obstacle of effective clinical application of EVs is the lack of disease-specific targeting efficiency to the sites of injury, especially to the CNS. It is well known that integrin a4ß1 is highly expressed by multiple pathogenic lymphocytes and mediates inflammatory cell homing into the CNS in MS. Currently, therapies targeting integrin a4ß1 have been used as a treatment for adult MS. Using ultra-high throughput one-bead one-compound (OBOC) combinatorial screening, we identified LLP2A as a high-affinity (IC50~2pM) and high-specificity targeting ligand against activated a4ß1 integrin. Furthermore, we showed that LLP2A is an a4ß1 integrin antagonist and can decrease inflammation. The overall goal of this proposal is to develop a new multi-component, multi-functional, EV based nanotherapeutic with improved targeting efficiency for MS. The treatment is comprised of unique PMSC-EVs that will be further engineered by surface modification with targeting ligand LLP2A. In this application, we hypothesize that conjugation of LLP2A onto the surface of PMSC-EVs (LLP2A-PMSC-EVs) can facilitate the targeting of PMSC-EVs to multiple activated pathogenic immune cell subsets, thus improving PMSC-EV targeting efficiency, tissue distribution, and ultimately therapeutic efficacy. We will leverage our lab’s established state-of-the-art bulk and single-EV characterization methods to evaluate ligand conjugation efficacy on EVs. The central objective of this study is to develop an effective approach to modify PMSC-EVs with LLP2A to efficiently target and modulate pathogenic lymphocytes to promote neuroprotection and remyelination. This study will facilitate future investigations into finding a novel bioengineering approach of stem cell-derived and cell-free regenerative therapy for MS.

NIH Research Projects · FY 2024 · 2024-09

PROJECT ABSTRACT Perinatal substance use in the U.S. has increased over the past two decades, with now over 8% of pregnant people endorsing recent use of a federally illicit substance (including cannabis, opioids, and cocaine). For newborns with prenatal substance exposure, multiple care practices during the birth hospitalization require special consideration: 1) approaches to newborn toxicology testing, 2) monitoring and management for newborn withdrawal symptoms, 3) counseling about human milk feedings, and 4) psychosocial support for families. Each of these aspects of care may shape the earliest parenting experiences of families, including their first interactions with the health system as parents. Given the issues of stigmatization and criminalization that surround perinatal substance use, the quality of care for substance-exposed newborns varies widely, with Black families and families living in poverty most likely to experience outcomes like child welfare services reporting and family separation. Further research is needed to identify hospital care practices that are readily implemented and that improve healthcare quality and equity for substance-exposed newborns. This will be an explanatory sequential mixed methods study conducted with the Better Outcomes through Research for Newborns (BORN) Network. Established by the Academic Pediatric Association (APA) in 2010, BORN is the first and only national practice-based research network that focuses on birth hospitalization care for healthy term and late preterm newborns. As a network of over 130 hospitals across 40 states and representing 10% of all U.S. births, BORN presents an ideal opportunity to study newborn care across diverse practice settings that span the legal and epidemiologic landscape of perinatal substance use. All study aims will be co-conducted with EMPOWER (Empowering Mothers, Providers, and Others to Weigh in as Experts in Research), a unique multi-state collaborative of patient stakeholders with lived experience of perinatal substance use and formal training in research methods. For Aim 1, we will develop, pilot-test, and disseminate a cross-sectional survey to all BORN Network hospitals (N=131) to delineate the range of care practices related to newborn toxicology testing, rooming-in, human milk feeding, and psychosocial supports for families, and we will identify common patterns of care. In Aim 2, we will retrospectively evaluate associations between hospital practices and patient- level outcomes among a purposive sample of 15 BORN hospitals (N=1,500 substance-exposed newborns). We will conduct multilevel regression, adjusting for clinical and contextual confounders, with a focus on differences by payor status and race. In Aim 3, we will assess the barriers and facilitators to implementing high quality and equitable care for substance-exposed newborns by conducting qualitative interviews and in-depth policy review at 6 hospitals. We will integrate the quantitative and qualitative findings from all 3 Aims to develop a set of recommendations for practices and written policies that can be disseminated for implementation across U.S. birth hospitals.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Infants and young children amass a vocabulary at a remarkable pace during the first 3 years. The size of vocabulary, and the rate of its growth predict future outcomes including academic achievement. Research on vocabulary development has largely focused on the cognitive processes underlying how young children learn new words, but we know remarkably little about the processes that support young children's retention of words over long delays to eventually become integrated into their lexicon. The examination of the neural processes supporting young children's retention will shed new light on contributing processes. We propose that the hippocampus enables learning and initial retention of the arbitrary association between words and their referents, but that additional cortical mechanisms in the anterior medial temporal lobe and medial prefrontal cortex are critical to respectively support longer retention and abstraction of meaning across multiple exposures. Further, we propose that opportunities to repeat and generalize which present in the child's environment are particularly likely to engage these cortical mechanisms in young children. Finally, we propose that neural activations capture processes underlying individual differences in vocabulary development. We developed novel functional Magnetic Resonance Imaging (fMRI) experimental paradigms that are appropriate for use in toddlers ages 25 to 35 months, a period during which toddlers learn hundreds of words. We will leverage targeted memory reactivation occurring during natural nocturnal sleep to elucidate neural mechanisms contributing to word retention and to predict vocabulary size and growth between 2 and 3 years of age. Relevance for Public Health: The acquisition of a vocabulary is critical for communication and understanding, and its size and growth predict various outcomes, including reading ability. Language acquisition is impaired in several neurodevelopmental disorders underscoring that a characterization of memory and vocabulary development is key to understanding adaptive functioning in various populations of children. This research will contribute to provide a stronger conceptual, empirical, and methodological foundation for examining populations of children who exhibit altered language development and memory difficulties due to a variety of conditions (e.g., Autism Spectrum Disorder, Down's Syndrome).

NIH Research Projects · FY 2025 · 2024-09

Personalized genetics is poised to revolutionize healthcare; however, despite advances, genetic prediction and portability of critical variants remains too limited for clinical use. Genetic prediction is error-prone when applied to individuals with genetic ancestries different from discovery cohorts, often predicting disease risk little better than random in non-European samples. We propose understanding an underlying cause of this loss of prediction accuracy by assessing the extent of GxG interaction effects across ancestries. Using an innovative approach, we tackle this statistically challenging problem by 1) modeling effect sizes of meQTLs on different ancestry haplotypes in an admixed African sample from the same population -- thereby controlling for GxE, and 2) using highly heritable genome-wide methylation phenotypes, affording us thousands of observations per participant rather than a single phenotype (e.g. presence of cardiovascular disease). Our study design allows us to assess whether the presence of ancestry-dependent interactions is a common factor in the variability of SNP effect sizes across populations. Outcomes of this grant include: generating a large genome-wide methylation dataset from 500 admixed South Africans, paired with underlying genome-wide DNA variation. We will further estimate the fraction of meQTLs with ancestry-specific effects and thereby comprehensively provide a snapshot of the frequency of GxG interactions in the human genome. These results will motivate investigation of GxG effects in a broader set of biomedical phenotypes and the extent to which they contribute to poor portability of polygenic risk across populations.

NIH Research Projects · FY 2024 · 2024-09

SUMMARY/ABSTRACT This proposal requests funds to purchase a BD FACSymphony S6 flow cytometric sorter to be housed in an active and established core, the Flow Cytometry Core at the California National Primate Research Center (CNPRC). The CNPRC is one of seven national designated primate research centers. Local, regional, and national investigators pursue translational research at CNPRC that is relevant to immunology, infectious disease, gene therapy and somatic cell genome editing, regenerative medicine and stem cell transplantation, cardiovascular disease, cancer, and neurodevelopment. The instrument is requested to support >20 current NIH-funded research projects from 6 Major Users and 11 Minor Users, as well as pending and future NIH- funded research, that spans basic, translational, and even clinical research. These projects include studies of therapeutic HIV vaccines, SIV pathogenesis, genome editing, retinal gene therapy, tissue-targeted AAV vectors, and even a phase-1/2a human clinical trial of a next-generation COVID vaccine candidate, for which mechanistic immunologic analysis will be performed at UC Davis. All these studies and indeed much of modern biomedicine requires the ability to separately study functions of specific cell types, or even individual cells. To perform such studies reproducibly and at an adequate quality level, furthermore, requires an instrument that is consistently checked and maintained, and one that should be under service contract. The BD FACSymphony S6 flow cytometric sorter will serve the increasing number of samples of our Users that are currently stressing a >20-year-old FACSAria flow sorter. The FACSAria was purchased using institutional funds and has been successfully maintained for an extraordinarily long serviceable lifetime, and was under service contract continuously until BD no longer offered the contract. However, the age and wide use of the older instrument impacts availability to the expanding NIH-funded research of our Major Users. Together, these constraints are hampering the progress of NIH- funded projects in their laboratories as well as efforts of their collaborators. The FACSymphony S6, with its greater selection of channels matched to those in our analytic cytometers, better robustness, more consistent maintenance, and higher laser power, will significantly advance our Major and Minor Users’ pursuits of novel mechanistic insights. We will integrate this instrument into an existing CNPRC core facility, the Flow Cytometry Core, which is maintained by UC Davis and CNPRC with significant infrastructure and oversight and advisory committees. The existing infrastructure will integrate the operation, maintenance and management of the proposed instrument which will, in turn, stimulate new research projects and innovations for many years.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract Unintended drug block of cardiac ion channels remains a major problem in drug development. The voltage-gated potassium channel KV11.1 also known as the hERG channel is a major drug anti-target that binds a diverse set of small molecule drugs. Drug-induced inhibition of the hERG channel reduces the critical repolarizing current IKr and thereby prolongs the cardiac action potential. Many drugs that bind the hERG channel promote deadly arrythmias like Torsades de Pointes while some hERG blockers present significantly lower proarrhythmic risk. Two hypotheses were proposed to elucidate this discrepancy: (1) preferential drug binding to the inactivated state of the hERG channel confers greater proarrhythmic risk and (2) simultaneous drug binding to other cardiac ion channels can ameliorate the risk associated with hERG channel block. Here, we present a state-specific molecular modeling assessment of drug binding to different conformations of the hERG and voltage-gated sodium hNaV1.5 and calcium hCaV1.2 channels. We have developed structural models of these cardiac ion channels in open and inactivated conformations and will perform all-atom molecular dynamics simulations to validate structural stabilities and assess ion conduction. Ligand docking was then performed using Site Identification by Ligand Competitive Saturation (SILCS), a pre-compute ensemble molecular docking technique. SILCS allows us to perform a high-throughput assessment of ligand binding affinities using molecular fragment energy maps derived from molecular dynamics simulations of state-specific ion channel models. Bayesian machine learning was used to provide improved correlation of SILCS-computed affinities with experimental data. Using SILCS multi-ligand docking we also estimated interactions of drugs with sex hormones in the hERG channel pore to assess a potential molecular mechanism for an increased proarrhythmia risk in females. We aim to use SILCS computed state-specific drug affinity data to inform multi-scale functional kinetic models of cardiac electrophysiology to estimate emergent drug effects on the cardiac action potential and heart rhythm. These modeling results will be validated by performing voltage-clamp electrophysiology measurements in vitro and compared to other published work. Upon completion of this project, the applicant will have refined expertise in computational modeling of ion channel-drug interactions and will have received training in experimental electrophysiology for the first time. Throughout the project, the sponsors will implement a training plan to strengthen the applicant’s knowledge of cardiac physiology and pharmacology while also broadening their scientific network, and improving the applicant’s scientific communication skills. This plan is tailored such that the applicant is prepared for a career as an academic scientist, investigating drug interactions with ion channels.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY 5.8 million individuals miss or delay medical care because of a lack of transportation. Persons with end stage kidney disease (ESKD) particularly depend on reliable transportation to dialysis sessions. In 2022, 84% of individuals newly diagnosed with ESKD initiated their treatment at an in-center hemodialysis (HD) facility, requiring they travel three times a week for HD. Yet 10% of patients with ESKD treated with HD miss dialysis at least monthly, and 35% miss treatment at least once every three months. Missing dialysis has significant individual and health system consequences, including increased emergency department visits, hospitalizations, and premature mortality and contributes to persistent racial and socioeconomic disparities in ESKD prevalence and outcomes. This Mentored Patient-Oriented Career Development Award (K23) will provide me with the training and mentorship needed to become an independent clinician-investigator and leader on transportation insecurity, social risk, and healthcare access for patients with ESKD. I am a family physician and medical anthropologist trained in qualitative and health services research and bring a unique perspective on how individual and community social context impacts healthcare. To achieve my long-term research and career goal of improving patients’ access to health care services and addressing longstanding health inequities, I need additional skills in (1) epidemiologic design and advanced quantitative analysis skills; (2) policy development and evaluation in transportation and ESKD, with emphasis on quality metrics; (3) clinical intervention development and implementation of future transportation solutions. I will accomplish the study’s goals through the following aims. Aim 1: Quantify social risk levels in a diverse group of patients with ESKD treated with in-center HD and associations with key HD outcomes in a prospective cohort study of 300 patients using the Accountable Health Communities (AHC) instrument. Aim 2: Assess the predictive validity of a novel transportation screening tool, the Transportation Security Index (TSI), in HD settings to identify patients most likely to miss HD appointments and compare its performance to the AHC’s transportation question. Aim 3: Identify facilitators and barriers to home dialysis as a component of addressing transportation insecurity in the ESKD population by conducting semi-structured interviews with patients, dialysis staff, and policy makers. The current proposal aligns with NIDDK’s priority area of developing interventions to integrate social and medical care to improve health equity and will break new ground by evaluating social risk within the ESKD population, with a focus on transportation insecurity.

NIH Research Projects · FY 2025 · 2024-09

Abstract Human eye development is an exquisitely balanced crosstalk of numerous signaling pathways that iteratively dictate the ebb and flow of this complex and highly physiologically active organ. Amongst the plethora of genetic mutations that can derail this sensitive programming, larger scale ocular malformations are encapsulated under a spectrum known as Microphthalmia (small eye globes), Anophthalmia (missing eye globes), and Coloboma (failed closure of the choroid fissure), collectively referred as MAC syndrome that affects approximately 1:10,000 live births. The following proposal highlights an anomalous index case of human bilateral anophthalmia and XX sex-reversal (BASR) syndrome. This anatomically male child was born with bilateral anophthalmia, a left cystic orbital mass, focal brain lesions, and is genetically female (XX). Their disease is the result of a de novo mutation on the long arm of their paternal X chromosome (Xq27). This region contains an extra copy of a large fragment (640k base pairs) from the long arm of chromosome 9 (9q21) which translocated during paternal meiosis I. The translocated fragment contained regulatory elements of an ion channel gene, TRPM3. Most fascinatingly, the exact point in which the translocation happened was at a 180 base pair palindrome on Xq27 that has been the causal site of mutation for a staggering number of other congenital disorders that affect, not just the eye, but numerous other tissue compartments or organ systems. These disorders involve the translocation of different autosomal fragments that land at this locus. To address BASR etiology, as well as molecularly unify all the disorders that map back to this translocation hotspot, we hypothesize that SOX3, a neighboring transcription factor, is ectopically activated within tissue domains that correspond to the translocated fragments. Additionally, we hypothesize that SOX3 in BASR was activated in the pigmented monolayer of cells in the back of the retina (retinal pigment epithelium; RPE) that is essential for photoreceptor health and ocular genesis. The following training plan seeks to test this hypothesized SOX3 etiology through both in vitro and in vivo modalities. By utilizing immortalized blood cells from the BASR patient, I aim to characterize the misfolded higher-order genetic landscape that comprises the Xq27 mutation through sequencing of closely associated regions (Hi-C). Additionally, induced pluripotent stem cells from the BASR patient will be induced to become RPE, during which I will gather and conduct serial analysis of the genetic expression of this process within a BASR context via single-cell sequencing. I aim to complement these efforts with characterization of a BASR mouse model, in which partially formed eye rudiments within these newly generated transgenic mice that ectopically express Sox3 will be immuno-detected and qualitatively assessed for any potential transdifferentiation.

NIH Research Projects · FY 2025 · 2024-09

Summary The overarching goal of the proposed work is to engage spousal caregivers of patients with dementia to plan for their own health emergency. Serious, unexpected health events such as hospitalization of a caregiver can become a major disruption to the caregiving relationship. Often the caregiver has not planned for such unexpected events where they may become ill. The project specifically focuses on the online development and testing of an Emergency Preparedness Toolkit (EPT). The EPT is designed for caregivers, in the event of an emergency, to provide a standby caregiver with detailed plans of how to care for the person with dementia. Interventions which support a caregiver are known to reduce caregiver burden and delay or offset the need to place a person with dementia in a care facility. Particularly challenging for some caregivers is the identification of a standby caregiver, especially if they do not have nearby family/friends. Previous work using the paper version of the EPT indicated that some caregivers had difficulty identifying a standby caregiver, recognizing the value of using the EPT and/or stalled by the amount of work needed for EPT completion. The online version of the EPT will provide tailored guidance to the caregiver and may increase the number of caregivers who identify a standby caregiver and are able to complete the EPT. Alternatively, caregivers may require coaching in addition to the online EPT format. Therefore, once the EPT is developed into an online format, spousal caregivers will be recruited and randomized to receive access to the EPT either with or without a coach. The first aim is to compare the two groups as to their ability to identify and engage a standby caregiver. Once the caregiver has successfully completed the online EPT they will they provide it to the standby caregiver. The second aim is to compare the change of spousal caregiver preparedness, confidence, and self-efficacy between groups. The third aim will examine the barriers and facilitators for spousal caregivers to identify and engage a standby caregiver for a person with dementia.

NIH Research Projects · FY 2024 · 2024-09

Project Summary: Regulation of immune responses in the lung is a critical determinant of health. While the immune system must respond to viral and bacterial pathogens, these responses must be finely tuned so as not to reduce organ function. Aberrant immune responses to viral pathogens in the lung can result in significant morbidity and mortality. Control of maladaptive host immune responses occurs through a number of mechanisms including input from the nervous system. Reciprocal communication between these two systems throughout the body has emerged as a mechanism to detect pathogens and coordinate host-protective immune responses and control inflammation. Although neuro-immune circuits in the lung are known to exist, the contribution of these to the regulation of inflammation caused by virus-derived pathogen-associated molecular patterns is unknown. Building on our preliminary data showing the regulation of lung inflammation by a unique neuro-immune circuit, the overall goals of this project are to precisely determine the role of vagal afferent neurons, the neuro-immune circuitry, signaling used, and how immune cell function is regulated. By using complementary and unique tools, the role of vagal afferent neurons and the processes these neurons evoke to suppress lung inflammation will be determined. To achieve this, we will use selective activation of vagal afferent neurons, perform identification of lung-specific vagal afferents, and confirm the role of β2AR and the source of catecholamines that regulate acute lung inflammation induced by a TLR agonist. Finally, we will assess if this novel anti-inflammatory pathway can be evoked to control inflammation during influenza infection (SA1). Understanding of the endogenous reflex triggered by lung inflammation will be performed using a combination of optogenetic silencing of the vagal afferents, pharmacological antagonists, and novel mice with conditional TLR7 deficiency in vagal afferents, to deduce if neurons sense viral components, and the mechanisms of immune regulation elicited. Finally, the role of sympathetic lung innervation will be assessed, and the requirement for specific neurotransmitter receptors tested (SA2). Together, these proposed studies will decipher the contribution and components of a novel lung anti-inflammatory pathway that limits acute lung inflammation.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY (ABSTRACT) With an increasingly aging and diversifying US population, the prevalence of Alzheimer’s Disease and related dementias (ADRD) is expected to rise. The expected increase highlights the importance to address modifiable risk factors to decrease the burden of ADRD and lead to better health outcomes. Obesity is a modifiable risk factor that continues to pose a public health concern. Mid-life obesity is considered a risk factor for ADRD, however late-life obesity appears to have either a null or protective association with ADRD. Potential explanations for these mixed findings are selective survival bias, competing risk of mortality, inverse causality, and BMI’s estimation of adiposity. These conflicting results affirm the need for a lifecourse approach to fully delineate the contribution of obesity at different ages as a risk or protective factor on ADRD and cognitive decline. This project will further evaluate the association between lifecourse obesity and ADRD and cognitive change, while addressing selective survival and competing risk biases that frequently impact findings from observational cohort studies on aging. Data will be leveraged from three ongoing well-characterized diverse cohort studies of long-term Kaiser Permanente members: the Kaiser Healthy Aging and Diverse Life Experiences Study (KHANDLE) (n=1712), the Study of Healthy Aging in African Americans (STAR) (n=764), and the LifeAfter90 Study (LA90) (n=999). These harmonized cohorts provide up to 60 years of data with repeated BMI and cognitive measures. BMI will be measured using height and weight collected from health records. Adolescent BMI will be any BMI readings between the years 10-20. Mid-life BMI will be from the ages 40-60 years, and late-life BMI will be from ages 65 years and above. Participants will be grouped into four categories based on their BMI: obese, overweight, normal, and underweight. Cognition will be measured using the Spanish English Neuropsychological Assessment Scale annually for KHANDLE and STAR and twice a year for LA90 participants. To address methodological concerns of selective survival and selection bias, Aims 1,2, and 3, will use inverse probability weighting. Aim 1 will examine different lifecourse pathways to evaluate the risk of adolescent, mid- and late-life obesity on cognitive decline. Aim 2 will address competing risk bias and evaluate the risk of mid- and late-life obesity with ADRD. Aim 3 will determine the mediating impact of lifecourse obesity on the association between gender and ADRD and cognitive decline to better understand the influence of lifecourse obesity on the well characterized gender differences in ADRD and cognitive decline. The results from this proposed study have potential public health impacts by addressing the role of a modifiable risk factor over the lifecourse on ADRD. These results will contribute to a better understanding of the epidemiology of ADRD and cognitive decline in diverse populations.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Botulism is a serious paralytic disease that affects both humans and animals which is caused by botulinum toxins (BoNTs). Animal botulism outbreaks in farm animals, in particular, not only affects animals but also result in financial losses for the industry and increases the risk of human exposure to BoNTs through animal products such as milk. To prepare for a response to the future outbreaks, a fast, sensitive, and reliable detection method for BoNTs is needed. The Endopep-MS method is one of the most promising methods that has been used for confirmation of human botulism by the Centers for Disease Control and Prevention (CDC) for years. However, a comprehensive validation study of the Endopep-MS for animal specimens and feed is lacking, and none of the veterinary diagnostic laboratories in the US have standard procedures for performing this method. To fill the knowleage gap, our team at the California Animal Health and Food Safety Laboratory (CAHFS), a founding member of VET-LIRN, is proposing a comprehensive validation study of the Endopep-MS for qualitative detection of BoNT/A, /C, /CD, /D, and /DC in animal specimens and feed. Our study has three specific aims as follows. Specific Aim #1: To validate the Endopep-MS for qualitative detection of BoNT/A, /C, /CD, /D, and /DC in animal specimens and feed. We will validate and compare the developed method to the standard mouse bioassay (MBA) using toxin-spiked samples and archived samples collected during natural botulism cases. The sample matrices include but are not limited to animal sera, liver, gastrointestinal contents, and feed. Specific Aim #2: To develop monoclonal antibodies for the toxin extraction step in the Endopep-MS method. Monoclonal antibodies are key for the Endopep-MS to achieve selectivity toward BoNT serotypes in samples. Since there is currently only one source of the monoclonal antibodies in the US, we will collaborate with Dr. Christina Tam at United States Department of Agriculture (USDA) and develop new monoclonal antibodies for the method. Specific Aim #3: To assess the effectiveness of polyclonal antibodies for the toxin extraction step in the Endopep-MS method. The limited source of monoclonal antibodies can delay research development and adoption of this method, an issue we have experienced ourselves. Alternately, polyclonal antibodies are commercially available but the ability of polyclonal antibodies for BoNTs extraction has not been completely explored. We will complete this knowledge gap by investigating capability of polyclonal antibodies for extraction of BoNT/C, /CD, /D, and /DC. If this project is funded and achieved, the lethal and time-consuming MBA that has been performed at CAHFS for close to 30 years can be avoided. Our laboratory, as part of the Vet-LIRN, will have the Endopep-MS ready to support botulism testing during outbreaks and large-scale animal feed emergency events.

NIH Research Projects · FY 2024 · 2024-09

ABSTRACT The US Environmental Protection Agency recognizes wildfire smoke as a significant source of particulate matter under 2.5 microns diameter (PM2.5) emissions, which are expected to increase because of climate change. The National Oceanic and Atmospheric Administration estimates that 200 million people in the United States live in counties affected by wildfire smoke conditions. Exacerbation of pre-existing respiratory conditions, including asthma and COPD, is associated with wildfire events. However, there is a paucity of data on whether wildfire smoke exposure can promote the development of chronic disease. Understanding the human health risks of wildfires is complicated by highly variable emissions from combined combustion of biomass and anthropogenic materials at the wildland urban interface. Further, growing epidemiological evidence suggests that particulate matter source and composition can be associated with more severe health outcomes. There is a compelling need to understand the toxicology of wildfire smoke exposures for public health, particularly in vulnerable populations such as young children. The goal of our revised R21 application is to address the significant knowledge gap by which inhaled wildfire smoke can elicit cellular injury in the respiratory tract of pediatric populations. We hypothesize that neonatal lung epithelium is more susceptible to inflammation and injury as a result of wildfire smoke PM2.5 exposure. We further propose that wildfire smoke PM2.5 exposure can elicit endoplasmic reticulum (ER) stress in lung epithelium, progressively leading to restrictive lung disease. Our hypothesis is based upon preliminary data with evidence of immune dysregulation and restrictive lung disease in adult rhesus monkeys exposed as infants to the 2008 Northern California Humboldt/Trinity County fires, as well as altered expression of lung ER stress markers and surfactant protein C (SP-C) in juvenile rhesus monkeys exposed to the 2018 Northern California Butte County “Camp” fire. We will test our hypothesis using wood smoke particles as a surrogate for wildfire smoke PM2. in primary cell or organoid lung epithelial cell cultures, followed by in vivo exposures using a neonatal mouse model. The Specific Aims of this revised R21 application will (1) Define the contribution of chronologic age on impact of wood smoke particle toxicity in respiratory epithelium; (2) Define the contribution of chronologic age on impact of wood smoke particle toxicity in alveolar epithelium; and (3) Investigate the effect of early life wood smoke particle exposure on induction of immune dysregulation and lung function decrements in the adult lung. To understand how the immune system and respiratory tract can be functionally and persistently altered by air pollutants, an essential step is to determine the cellular mechanisms of exposure in vulnerable populations such as young children. Our findings will inform on the unique toxicological aspects of air pollution exposures resulting from wildfires and lead to development of biomarkers and preventative measures to limit the adverse health effects of restrictive lung diseases.

NIH Research Projects · FY 2024 · 2024-09

UC Davis Cardiovascular Symposium Systems Approach to Understanding Cardiovascular Disease -Ion Channels, Ca2+ regulation and signaling- Cardiovascular diseases, including hypertension, heart failure, arrhythmias and stroke, are the number one killer in the developed world. In order to develop better and more effective therapies to treat cardiovascular diseases, it is critically important for scientists and physicians to obtain in-depth and accurate understanding of the mechanisms underlying heart and vascular function and dysfunction. In recent decades, researchers studying heart and vascular diseases have been accumulating more and more experimental data from the molecular, to cellular, to tissue and organ levels. However, there is a critical need to integrate these data into mechanistic and quantitative models to understand emergent properties of complex biological systems, such as arrhythmias and vasospasms that are often counterintuitive due to non-linear dynamics interactions. Moreover, just from a fundamental view, it is important to understand how different cells of the cardiovascular system may interact with each other in physiology and disease. Here, we propose to take the necessary step forward to integrate experimental data into quantitative models that enable the use of mathematical tools and computational power to understand the complex interactions of the cells and molecules in the cardiovascular system. The unique design of this conference series is to combine experimental study and mathematical modeling to achieve in- depth understanding of the dynamic systems that control cardiovascular function in health and disease. The proposed interdisciplinary conference is the 8th in this series. The last installment of the conference integrated, for the first time, studies in cardiac and vascular tissue. The previous conferences have received overwhelmingly positive evaluations from attendees and resulted in high-impact publications. The proposed conference will combine experimental and modeling studies in the field of cardiac and vascular physiology and focus on in-depth comparison of the cardiac and vascular smooth muscle function, with emphasis on ion channels, cellular Ca2+ regulation, and signaling. The aims of this conference will be on (1) summarizing the current state of research in the focus area, (2) identifying consensus and controversy that warrant more investigation, and (3) exchanging ideas, data, and information among the experimentalists and modelers to facilitate interdisciplinary collaborations. As previously done, the conference results will be summarized in the form of a comprehensive review paper, which will be published in a leading scientific journal that have broad impact on the research community.

NIH Research Projects · FY 2025 · 2024-08

Simultaneous two-photon imaging and two-photon manipulation of neural activity in freely- behaving mice The ability to record and manipulate neural activity in freely-behaving mice is a key step to understand the neural circuits under natural behavior. One-photon miniscope has become a mature technology and it can perform calcium imaging and optogenetics simultaneously while the mice freely behave. Though it has enabled many new discoveries in neuroscience, the ability to image and manipulate neural activity deeper into the tissue and in higher spatial specificity can further advance the field and enable studying many new questions. Compared to one-photon, two-photon techniques can access much deeper tissue and have a much higher spatial specificity. However, it has been challenging to integrate high performance optics into a compact footprint miniscope to perform simultaneous two-photon imaging and two-photon optogenetics, due to the excessive challenges in optical and mechanical design. Here, we propose a new miniscope that can simultaneously perform two-photon calcium imaging and two- photon optogenetics in freely-behaving mice. We innovatively integrate two different beam forming techniques in the miniscope for the imaging beam and optogenetics beam. This unique combination enables a very compact mechanical design and a high optical performance. Crucially, we will achieve a high-spatiotemporal-resolution in imaging, and patterned stimulation in optogenetics where a group of user-selected neurons could be simultaneously photostimulated. This allows us to manipulate the ensemble activity while monitoring the response of the neural circuit, all in cellular resolution. The entire device can have a dimension <~14x14x25 mm3 and a weight <3 g, suitable to be mounted on the skull of freely-moving mice. Furthermore, we will custom design and manufacture the optics to support a large field of view of 400 µm in diameter. This allows us to access a large amount of neurons. Finally, the focal depth of both beams could be controlled independently so we could image and manipulate the neural activity across a 3D brain volume. The success of this project will create a two-photon miniscope that can simultaneously image and manipulate neural activity, both in cellular resolution, high temporal resolution/specificity and over a large 3D volume deep in the brain tissue, in freely-behaving mice. The proposed miniscope could enable new research that is previously not possible, such as investigating the neural circuits of 2D navigation and social behavior. Our miniscope will greatly benefit the neuroscience community, and be readily deployed to many research labs. While we will design and test the miniscope for mice, its application could be extended to rats and non-human primates in future.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract Type 1 diabetes (T1D) in childhood may cause neurocognitive deficits which worsen over time. Declines in cognitive function may lead to suboptimal T1D management, resulting in a vicious cycle that increases risk for poor glycemic control and life-threatening complications. New devices for T1D management such as automated insulin delivery systems reduce glycemic variability, but optimal use requires advanced understanding and cognitive functioning. Therefore, understanding how to prevent neurocognitive decline is critical. Despite the importance of this issue, the mechanisms underlying neural injury caused by T1D are not well understood. Exposure to diabetic ketoacidosis (DKA) at T1D onset and repeated DKA episodes, chronic hyperglycemia, and severe hypoglycemia have all been hypothesized to play a role. In the proposed multi- center study, we will conduct a longitudinal cohort study of children with T1D and a comparison group of children without diabetes, both from diverse racial, ethnic, and income backgrounds, to characterize associations between T1D-related factors and neurocognitive outcomes and to identify modifiable mechanisms and actionable targets for ameliorating cognitive declines. We will enroll 800 children 6 to 11 years-old with new onset of T1D (half with and half without DKA at onset) and 200 children without T1D. This cohort will be comprehensively evaluated at enrollment and followed for a minimum of 1.5 years to determine the extent to which DKA at onset predicts cognitive differences over time (Aim 1), how glycemic variables (mean HbA1c, time in range, mean amplitude of glycemic excursions, frequency of hypoglycemia) predict cognitive differences over time (Aim 2), and to assess interactive effects of DKA exposure and glycemic variables. We will also investigate whether DKA and hyperglycemia are associated with changes in serum inflammatory / neuroinflammatory markers and brain structure via magnetic resonance imaging (MRI, Aim 3). Participants will complete a detailed Baseline assessment (3-6 months after T1D diagnosis) and follow up assessments (yearly after the baseline assessment). Comprehensive panels of inflammatory markers and measures of gene expression will be used to characterize the inflammatory/neuroinflammatory state over time. MR diffusion tensor imaging will be used to assess brain microstructure (fractional anisotropy) and MR spectroscopy will evaluate brain metabolic alterations related to neuronal health and density. Together, these results will provide information to develop a mechanistic model for T1D-associated brain injury on which to base future interventional trials. In sum, this project will characterize the impact of DKA and glycemic variables on cognitive functioning in children with T1D and will provide insights into how these factors may lead to neural injury and cognitive decline. By identifying risk factors for cognitive deficits, determining optimal glycemic ranges to prevent declines in cognition, and pinpointing avenues for prevention and intervention, we will gain the foundation necessary to change clinical practice and improve cognitive function in people with T1D.

NIH Research Projects · FY 2025 · 2024-08

The goal of this project is to understand reservoir effects and curative potential of a promising CCR5-depleting bispecific antibody (bsAb) given with days of ART initiation to infants with either limited or robust pre-ART viral replication. The agent is a bsAb with one arm directed against CD3 and the other against CCR5, which approximates cytotoxic T cells to CCR5-expressing cells. These cells form a key part of the HIV reservoir in ART-treated people and have been eliminated in notable cases of HIV cure. Our preclinical studies revealed >50% remission in simian immunodeficiency virus (SIV)-infected infant animals after bsAb treatment. We hypothesize that these effects were achieved through a combination of true reservoir depletion and removal of SIV target cells required for rebound. In this R01 application, we propose to improve the manufacturing and safety of this bispecific antibody and then rigorously test the two proposed mechanisms of action. We will screen early-infected infants to identify those with either (i) limited viral replication and the potential for cure, or (ii) robust replication and likelihood of establishing measurable reservoirs that can be followed during and after bsAb treatment. These two groups will be separately studied in order to isolate and measure reservoir effects and the potential for cure. Hypothesis: Highly purified CD3/CCR5 bsAb, given within days of ART initiation, safely achieves significant SIV reservoir depletion from infants, which is sufficient to induce durable remission in those with restricted reservoirs or can act synergistically with bnAb to achieve remission in animals with larger reservoirs. Aim 1. Test CCR5 depletion and side effects caused by CD3/CCR5 bsAb produced as a CrossMab, compared to controlled Fab-arm exchange (cFAE). Aim 2. Demonstrate the potential for infant SIV cure via CCR5 depletion near the time of ART initiation, when viral replication and reservoirs are limited. Aim 3. Measure the interference with reservoir establishment that is achievable by CCR5 depletion, bnAb, or the two combined in early-treated infants with robust viral replication.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Liver cancer is a major cause of cancer-related deaths within the United States, being the fifth and seventh leading cause of cancer deaths among men and women, respectively. There is a high demand for new and more effective therapeutics for the treatment of liver cancer, especially for the most common hepatocellular carcinoma (HCC), with an improved understanding of vital regulatory factors underlying HCC cell metabolism essential for tumor progression. Genome-derived noncoding microRNAs (miRNAs or miRs) have been revealed as critical elements to control posttranscriptional gene regulation, and restoration of liver-enriched, oncolytic miRNAs (e.g., let-7-5p isoforms, miR-122-5p, and miR-148a-3p) lost or downregulated in HCC cells represents a new therapeutic strategy. However, current miRNA functional and experimental therapeutic studies are limited to the use of miRNA mimics chemo-engineered in vitro and comprised of extensive and various types of artificial modifications, which are totally different from natural miRNA molecules produced in vivo. This is also in sharp contrast to protein research and therapy in which bioengineered or recombinant protein agents produced and folded in vivo, rather than synthetic polypeptides or proteins made in vitro, have been used and found enormous success. To overcome this barrier, the PI has recently developed a novel RNA molecular bioengineering platform technology, based upon specific hybrid tRNA/pre-miRNA molecules identified in the PI’s lab as carriers, to achieve high-yield and large-scale, in vivo fermentation production of true biologic or bioengineered RNA (BioRNA) molecules for basic and translational research. Our following studies have demonstrated that miRNA (e.g., let-7c-5p) is selectively released from BioRNA “prodrug” in human HCC cells to regulate target gene expression (LIN28B) and control cellular processes (tumorsphere formation), and liposome-polyethylenimine (LPP) nanocomplex is superior to in vivo-jetPEI to deliver BioRNA into orthotopic HCC tissues and inhibit tumor growth in mouse models. Our further efforts have led to the identification of proper human tRNAs to couple with human hsa-pre-miRNAs as new carriers to offer humanized BioRNAs to target HCC. In addition, humanized BioRNA/miR-148a-3p is precisely processed to miR-148a-3p in human HCC cells to modulate specific amino acid and glucose transporter expression towards the control of aminolyses and glycolysis. Given these exciting preliminary findings, we hypothesize that novel, HCC-targeted, humanized BioRNAs can be bioengineered and used to dissect HCC nutrient homeostasis and tumor metabolism. To test the hypothesis, we proposed to (i) design, clone, express, and purify a focused group of novel humanized BioRNAs for the inhibition of HCC cell viability (Aim 1), (ii) delineate the mechanistic actions of BioRNAs in the control of HCC cell metabolism (Aim 2), and (iii) establish the effectiveness and safety of candidate BioRNAs in the modulation of HCC tumor metabolism and progression in vivo (Aim 3). Successful completion of this project will offer a set of one-of-a-kind, biologic, HCC-targeted RNA molecules as research tools, establish new roles for miRNAs in the control of HCC cell metabolism, and build a solid foundation for the development of new remedies to improve the treatment of lethal HCC.