University Of California-Irvine

NIH Research Projects · FY 2024 · 2019-07

): Nearly 100 million Americans were afflicted by at least one of more than a thousand neurological diseases, according to data for 2011. The risk of dementia increases exponentially with age, with disproportionate effects on blacks and Hispanics. Finding effective treatments for neurological disorders and strokes requires fundamental knowledge of the nervous system and the participation of a diverse workforce to enhance our overall creativity. In 2017, the U.S. Department of Education named the University of California, Irvine, (UCI) as a Hispanic-serving institution, meaning that one-quarter of the undergraduate student body identifies as Latino and that half of all students receive financial aid. More than 50 UCI faculty conduct research in neurosciences, primarily in the Departments of Neurobiology and Behavior (NBB), and Anatomy and Neurobiology. NBB, which was established in 1964, was the first neuroscience department in the world (five years before the formation of the Society for Neuroscience) and it is ranked among the top by the National Research Council. Over the last 18 years, the UCI Minority Science Programs (MSP) has developed innovative interventions to improve the academic excellence and to increase the number of underrepresented minority (URM) undergraduates being trained as the next generation of biomedical researchers. The objective of the program Broadening Research Achievement in Neurosciences (BRAiN) for a Diverse Workforce is to facilitate participants’ career advancement from community college to UCI and from college to Ph.D. programs in neurosciences. The measurable objective is to increase by three-fold the number of URM undergraduates entering Ph.D. programs in neurosciences each year. Participants will spend two years being mentored and conducting research continuously. During the sophomore year, participants will join the MSP training laboratory, which is dedicated to developing original research projects while providing a nurturing and stimulating environment for URM students who have not taken upper division classes. Subsequently, participants will join one of the UCI laboratories dedicated to neurosciences. Participants will take courses in neurosciences, scientific writing and training in the responsible conduct of research. At the end of the program, participants will have attended 30 research talks by neuroscientists, developed strong quantitative reasoning skills (including computer programming), presented their research findings at two national conferences, participated as co-authors on a paper based on their research experience and gained admission to Ph.D. programs in neurosciences at top universities.

NIH Research Projects · FY 2026 · 2019-05

PROJECT SUMMARY An urgent child policy question facing the U.S. concerns how much and in what way to invest in early care and education, particularly pre-kindergarten (pre-k). Rigorous evaluations of pre-k programs within states have produced positive, negative, and null findings that are difficult to reconcile, necessitating an approach that can integrate data across time and contexts. We ask whether variations in investments by the 50 states across the past 20 years are associated with better child health, achievement, and family outcomes. Our study will build on our previous work in North Carolina (NC), which examined the effect of variation in pre-k funding to counties across years on child outcomes using econometric analyses of panel data. Our first aim is to evaluate whether the beneficial impacts of investments in public pre-k generalize across the nation and with other estimation approaches. We will use this rigorous method at the national level by compiling the first state-by-year-level dataset that integrates pre-k funding, pre-k program features, state characteristics, and child and family outcomes (e.g., NAEP scores, anxiety, disability, maternal employment), applying a state- and year-fixed effects approach. We also incorporate new NC data for an instrumental variables approach, using the interaction between a family’s proximity to the nearest NC Pre-K center and funding for pre-k as an instrument for pre-k attendance—providing innovative causally-informative evidence of pre-k effects. The second aim is to determine whether statewide investments in pre-k promote children’s well-being across different subgroups. In our nationwide study, we will test whether state investment levels have differential impact on subgroups of children, and in NC, we will test whether our new analytic approaches demonstrate beneficial impacts on child achievement for all populations of children. The third aim is to examine whether features of state pre-k programs and children’s environments are associated with differential impact of states’ investments, which we test through moderation analyses of carefully collected pre-k implementation data in each state and year. The fourth aim is to examine whether prior NC Pre-K exposure protected children from the adverse impact of economic shocks and natural disasters, by contrasting academic achievement for NC Pre-K attenders with a matched group of non-attenders during pre- and post-economic crises, and from counties affected by hurricanes, floods, and snowstorms. Together, this work will inform child development theory about the enduring impact of early education experiences and optimal policy strategies for promoting positive learning outcomes.

NIH Research Projects · FY 2025 · 2019-04

The broad, long-term objective of the UC Irvine Skin Biology and Disease Resource-based Center is to promote significant and innovative discoveries in regulatory mechanisms in skin biology and disease through cross-fertilization between multiple disciplines, integrated through systems biology. The mission of the Administrative Core is to centralize the leadership and operations of the Center, and to provide the administrative support needed to meet the broad, long-term objective of the Center. The Administrative Core will exert leadership for skin biology research at UCI and nationwide and oversee outreach and enrichment activities of the Center that will expand the community of scientists pursuing NIAMS-mission research. The Center is led by Director Bogi Andersen and Associate Directors Anand Ganesan and Arthur Lander, with input from the Executive and Advisory Committees. The Administrative Core will manage funds and other Center activities; oversee communications; set milestones and oversee surveys and other methods of evaluations; and oversee Community Engagement efforts. The Administrative Core also runs a series of programs of enrichment, including weekly Skin Club, monthly seminar series, and yearly symposia, as well as several mechanisms for seed grant and other funding types that advance the specific aims of the Center in the upcoming cycle. In the first cycle, the Center Administration successfully enhanced collaborations and synergy among research community members and attracted new researchers to the study of skin biology and disease. The initial funding of the Center spurred a marked increase in technology development, innovative research publications, and grant funding. By servicing the various needs for the three resource Cores and the P30 research community members, the dedicated effort and strong support of this centralized Administrative Core provides added value and enhances the synergy of the research community of the UC Irvine P30 Center.

NIH Research Projects · FY 2025 · 2019-04

Hippocampal sclerosis of aging (HS-A) and Limbic predominant age related TDP-43 encephalopathy neuropathologic change (LATE-NC) are important degenerative pathologies that are closely related but distinct. Around 20% of older age dementia is attributable to these two conditions. HS-A is diagnosed when there is disproportionate atrophy in a brain structure called hippocampus and LATE-NC is characterized by abnormality of an essential protein, TDP-43. Clinically, both pathologic changes mimic Alzheimer’s disease pathology and present with memory problems. Despite their importance, none of the pathologies can be diagnosed during life and can only be found by examining the brain after patients die. Moreover, both HS-A and LATE-NC frequently co-occur with Alzheimer’s disease pathology. This makes their diagnosis even more challenging. This is especially important in an era that specific treatments for these protein abnormalities are being developed. Therefore, there is significant unmet need for discovering ways to diagnose them during life. In the current cycle of the grant, we showed that our quantitative method is more informative than standard pathologic measures in identifying relationships between HS-A and clinical impairment and presence of other pathologies. We also found that clinical features of HS-A and LATE-NC are remarkably similar to Alzheimer’s disease. We found that MRI detects atrophy of hippocampus in HS-A, and this is an early finding making MRI a suitable tool for HS-A diagnosis. We have also found indications that autoimmunity plays a role in HS-A. The objective of this proposal is to advance our HS-A discoveries and to start new investigations on pathological assessment and diagnosis of LATE-NC in an ethnically diverse group of participants. Inclusion of ethnically diverse participants will allow for studying the prevalence and impact of these important pathologies in a representative sample of our population. The study will use the resources of four ongoing studies of older individuals across two Alzheimer’s disease research centers at University of California (UC), Irvine and UC Davis, The 90+ Study, and LifeAfter90 Study. In aim 1, we will replicate our HS-A results in a new and ethnically diverse group of participants that will validate our methods. We will also test the hypothesis that quantitative assessment of LATE-NC will identify novel associations with cognitive impairment and other brain pathologies. In aim 2, we will test the hypothesis that brain MRI and glucose PET scan, can identify features that are specific to HS-A and LATE-NC. In aim 3, we hypothesize that blood markers of autoimmunity and inflammation are related to HS-A and LATE-NC respectively and measuring TDP-43 and progranulin in blood, can serve as markers of LATE-NC. Successful completion of this proposal, we lead to major advancement in the diagnosis of two important dementia related pathologies that cannot be diagnosed during life. This will not only enable realistic assessment of the effectiveness of Alzheimer’s therapies but will also pave the way for clinical trials of HS-A and LATE-NC.

NIH Research Projects · FY 2025 · 2019-04

Project Summary Neuronal Kv7 (KCNQ) and Kv1 (KCNA1) subfamily voltage-gated potassium (Kv) channel loss-of-function causes developmental epileptic encephalopathy, ataxia, hereditary spastic paraplegia (HSP) and addiction. In this project we focus on the mechanistic basis and potential therapeutic utility of plant-derived diterpenes and hydroxybenzoic acids that are extremely well tolerated in rodent and human safety studies, cross the blood- brain-barrier, and which we recently discovered to be potent, efficacious openers of neuronal Kv7 and Kv1 channels. Addiction and epilepsy represent major health burdens in the US and globally, and new approaches to their treatment are desperately warranted. Episodic ataxia and HSP are relatively rare and therefore can especially benefit from therapeutic development in academia; in turn, treatments developed for episodic ataxia can inform development of therapies for epilepsy and potentially other forms of ataxia. In this project we take a multidisciplinary approach incorporating molecular dynamics (MD) simulations, site-directed mutagenesis, electrophysiology, and in vivo testing in existing and novel mouse models. In Aim 1, we will pursue the molecular basis of action and isoform selectivity of the diterpene carnosic acid (CA), which we recently discovered to be a highly efficacious and isoform-selective Kv7.3/5 channel opener, and to Kv7.3/5-dependently inhibit cocaine- seeking behavior in mice. We now intend to delineate how CA opens Kv7.3 but not Kv7.2 using all-atom MD simulations, mutagenesis and electrophysiology. We propose similar studies for gentisic acid (GTA), a hydroxybenzoic acid we recently found to be the most potent known opener of Kv7.3 and Kv7.2/3. The studies are translationally highly significant because of the potential for CA to treat psychotropic drug addiction, and for GTA as a lead anticonvulsant compound. In Aim 2, we will determine the molecular basis of action of pisiferic acid (PA) and gallic acid (GA), another diterpene and hydroxybenzoic acid, respectively, that we found to be novel Kv1.1/Kv1.2 channel openers that can reverse the effects of Kv1.1 and Kv1.2 mutations that cause ataxia, epilepsy and HSP. We will use experimentally validated MD simulations of entire Kv1.1 and Kv1.2 channels in model neuronal lipid membranes, to determine how PA and GA achieve efficacy, potency and selectivity. Extensive preliminary data support an exciting paradigm in which PA co-opts the Kv1.1 voltage sensor to act as a ligand-binding/gating domain. We will also test the ability of PA and GA to rescue in vitro a broader panel of ataxia, epilepsy, and HSP-linked Kv1.1 and/or Kv1.2 mutants. We have already discovered that PA reverses EA1 in vivo in our new mouse model of Kv1.1-linked Episodic Ataxia Type 1 (EA1). In Aim 3, we will conduct in vivo testing of the therapeutic efficacy of PA and GA in mouse models of EA1 and Kv1.2-linked ataxia/epilepsy/HSP, and PK studies. The project will uncover transformative mechanistic paradigms for small molecule opening of voltage gated ion channels in general, whilst also providing a complete picture of the mode of action of four safe, efficacious, selective and/or potent Kv channel openers with true therapeutic potential.

NIH Research Projects · FY 2026 · 2019-02

Project Summary Voltage-gated potassium (Kv) channels, essential for cellular electrical activity, are generated by tetramers of pore-forming α subunits, often in complexes with other, non-pore-forming β subunits and other protein classes. Sodium-coupled solute transporters provide a mechanism for transport of water-soluble ions, neurotransmitters, vitamins, sugars and other small molecules across cell membranes and against the electrochemical gradient. In prior award cycles we discovered that Kv channels form physical complexes with sodium-coupled solute transporters and we defined multiple modes of co-regulation in these “chansporter” complexes, establishing a new class of cellular signaling hub. Concomitant with this work, we discovered a range of novel small-molecule modulators of Kv channels and chansporter complexes, including synthetic compounds and plant metabolites, some with therapeutic potential. In this latest cycle, we propose to pursue both these fields of study, focusing primarily on the Kv1 (KCNA) and Kv7 (KCNQ) Kv channel families, disruption of which causes disorders as diverse as ataxia, cardiac arrhythmia, diabetes, achlorhydria, hypothyroidism, and epilepsy, and the transporters with which they interact. We will build on our prior work and preliminary data that include novel chansporter complexes, novel modes of Kv channel chemosensing in chansporter complexes, and screening results revealing abundant new Kv channel modulators from plants. We have established two new, unpublished transgenic rodent lines for this project that will facilitate study of new therapeutic approaches to treat Episodic Ataxia 1 (EA1) (a mouse model), and of the precise roles in vivo of KCNQ5 and KCNQ5-transporter complexes (a Kcnq5 knockout rat line). We use a highly integrated approach to investigate the molecular mechanistic bases for channel and chansporter biology and pathophysiology, drawing from our long experience in studying molecular basis of biology and disease in multiple tissues, cellular electrophysiology, transport and radioligand assays, transcriptomics, various imaging modalities, structure-function studies, and biochemical techniques. In the next five years, we aim to address several critical knowledge gaps, pursuing the following novel research directions: (1) Molecular mechanistic studies of new and known channel-transporter complexes to dissect novel forms of co-regulation and signaling; (2) Channel/transporter-active small molecule discovery from plants, drawing from our completed dual-target screen of 1444 plant extracts; (3) elucidation of novel roles for KCNQ5 in the vasculature and brain; (4) in vivo testing of the first compounds known to directly rescue EA1-linked Kv1.1 sequence variants. Our overarching goals are to uncover new chansporter complexes and their roles in vivo, enhance understanding of Kv channel biology, and discover novel and therapeutically relevant channel/transporter-targeted small molecules.

NIH Research Projects · FY 2025 · 2019-02

PROJECT SUMMARY Breast cancer susceptibility gene 1 (BRCA1) is a tumor suppressor gene best known for is function in DNA repair. Early studies suggested that genome instability within the mammary epithelial system of BRCA1 mutation carriers (BRCA1+/mut) promotes a cascade of cell autonomous, genetic events ultimately giving rise to cancer initiation within a population of luminal epithelial progenitor cells. However, a major gap in knowledge is whether germline BRCA1 mutations lead to cell state aberrations within the cellular microenvironment surrounding the epithelium, which may act in trans to promote pre-cancerous changes in the epithelial system thereby promoting breast cancer initiation. In this renewal, we will expand on the provocative findings from our currently funded project, which identified dramatic precancerous changes in both breast epithelium and surrounding microenvironment of human BRCA1+/mut. Our work identified the expansion of epithelial cells with basal-luminal intermediate (BLI) phenotype as well as a population of epithelium-surrounding precancer-associated fibroblasts (preCAFs) expressing copious amounts of matrix metalloproteinase 3 (MMP3) and immunoregulatory factors (e.g., neutrophil-attracting CXCL8). Based on our progress and preliminary data, we will address the hypothesis that preCAFs emerge in a cell-intrinsic manner due to fibroblast-specific BRCA1 deficiency leading to an aberrant form of fibroblast differentiation. In this context, we will test whether cell-intrinsic NFkB activation in BRCA1+/mut acts as a central signaling node towards the preCAF state. We will also determine whether BRCA1mut fibroblasts establish a precancerous immunosuppressive microenvironment during breast cancer initiation through MIP-2, a mouse homologue for CXCL8. Addressing these major gaps in knowledge about the function and origin of preCAFs may pave the way towards novel therapeutic approaches to block the cancer-promoting function of preCAFs or reverse their phenotype as precision cancer prevention strategies.

NIH Research Projects · FY 2025 · 2018-08

PROJECT SUMMARY/ABSTRACT Tuberculosis preventive therapy (TPT) is known to reduce the incidence of tuberculosis (TB) among people living with HIV (PLHIV) and is considered a core service of National AIDS Programs. TPT coverage has been steadily increasing with the availability of short-course regimens such as 12 weeks of once-weekly isoniazid- rifapentine (3HP). However, treatment completion in programmatic settings remains sub-optimal, highlighting the need to identify feasible facilitation strategies that can maximize the impact of the considerable investments in TPT scale-up. Previously, in a single-center randomized trial in Uganda, we demonstrated that facilitated self-administered therapy (SAT) resulted in high levels (92%) of treatment completion, was preferred by most PLHIV and substantially reduced health system costs relative to directly observed therapy. Further evidence is now needed on whether the facilitation components are necessary and feasible to implement across a broad array of HIV clinics to guide TPT scale-up efforts. Building on our prior work, the overall objective of this proposal is to compare the effectiveness and implementation of simplified facilitated SAT to routine SAT for 3HP delivery. Simplified facilitation will include three components from our previous trial designed to address key barriers to 3HP completion: enhanced counseling, interactive voice response dosing reminders/adverse event check-ins, and 99DOTS-based adherence monitoring/support. Routine SAT will include enhanced counseling alone. Our central hypothesis is that simplified facilitated SAT will result in higher levels of completion than routine SAT and be cost-effective. In Aim 1, we will test our hypothesis by conducting a highly pragmatic cluster-randomized trial among 3400 PLHIV at 20 HIV clinics in three districts of Uganda to compare completion of 3HP using simplified facilitated SAT vs. routine SAT. The selected HIV clinics represent diverse levels of the healthcare system, geography (rural vs. urban/peri-urban location) and experience with 3HP delivery. In Aim 2, we will employ a mixed methods approach to assess the reach, adoption and implementation of facilitated SAT components, and whether or not they modified targeted barriers. Last, in Aim 3, we will collect data on the cost of simplified facilitation of 3HP from the health system and societal perspectives to estimate the incremental cost- effectiveness of simplified facilitated SAT as a function of key facility-level characteristics. Investments in scale-up of 3HP – the most promising intervention for TB prevention available today – will not achieve the desired impact unless 3HP can be delivered effectively and in a person-centered fashion. Our proposed studies will clearly demonstrate whether facilitation components are required to achieve high completion, feasible to implement across a variety of clinic settings, and cost-effective. These data will inform decisions as to whether simplified facilitation should be budgeted by HIV/AIDS programs as part of 3HP scale- up initiatives in high TB-burden settings throughout Africa and worldwide.

NIH Research Projects · FY 2026 · 2018-07

Summary The study of mammalian immune cells and their interactions with tissue in situ is critical for understanding how they regulate processes ranging from wound healing to autoimmune disease initiation to cancer and for designing better therapeutic strategies to treat these prevalent conditions. Intravital multiphoton microscopy (MPM) combined with a rich repertoire of fluorescent reporter mouse models and in vivo cell and tissue labeling techniques have made it possible to visualize immune cell-tissue interactions at a subcellular level in skin and other organs. However, there are significant differences in the structure and immune milieu of human skin that limits the translatability of these findings to the human cutaneous immune response. Our group has recently developed a fast large area multiphoton exoscope (FLAME), a unique imaging platform optimized for efficient clinical skin imaging to rapidly generate macroscopic images (mm to cm-scale) with microscopic resolution (0.5- 1µm) based on label-free molecular contrast (fluorescence intensity and lifetime). In this application, we leverage our extensive experience in MPM technology development and clinical imaging of more than 400 patients over the past several years to develop the first MPM-based clinical device (iFLAME) as a research imaging tool optimized for, and dedicated to, in vivo label-free imaging of immune cell populations and their dynamics in human skin. In Aim 1, we develop iFLAME as a clinical research tool for efficient in vivo label-free imaging of dermal cell populations and their dynamics in human skin. This work involves development of detection and analytic approaches as well as optical and computational methods to enable rapid fluorescence lifetime detection and analysis necessary to automate measurements of the cellular morphological and metabolic signatures. In Aim 2, we validate iFLAME performance by demonstrating in vivo characterization of immune cells in normal and inflamed human skin. In Aim 3, we develop quantitative morphological and metabolic MPM imaging endpoints to assess immune infiltrates and their dynamics in human skin in the context of monitoring wound healing. This work represents the first attempt to use intrinsic sources of MPM contrast to image, identify, and quantify key immune cells in human skin in vivo based on their optical signatures and migratory behavior. Our long-term goal is to develop iFLAME as a clinical research tool for rapid, label-free imaging of immune cells in skin based on cellular morphologic and metabolic imaging endpoints. These can be used to better understand, evaluate and optimize wound healing, autoimmune skin diseases and therapeutic responses.

NIH Research Projects · FY 2026 · 2018-07

IMPACT OF CANNABINOIDS ACROSS THE LIFESPAN (ICAL): SUMMARY Teenagers use cannabis more than any other recreational drug. Their developing brains may also be especially vulnerable to its effects, as epidemiological and experimental evidence suggests that frequent cannabis use in adolescence is associated with impairments in cognitive and affective functioning that continue in adult life. Excessive stimulation of the endocannabinoid (ECB) system – the target of cannabis’s intoxicating constituent, Δ9-tetrahydrocannabinol (THC) – is a plausible but still poorly understood mechanism for the lasting consequences of cannabis use. The NIDA Center Impact of Cannabinoids Across Lifespan (ICAL), whose renewal is proposed in this revised application, combines molecular, synaptic, and behavioral approaches to determine whether adolescent THC exposure (‘ado-THC’) alters ECB signaling in a persistent manner thus causing impairments in adult brain function and behavior. In the first funding period, which lasted 4 years, we made substantive progress on our research and service goals. Research: (1) We systematically characterized pharmacokinetics and metabolism of THC in adolescent and adult mice and rats of both sexes and validated a THC treatment protocol that models daily cannabis consumption (an increasingly common use pattern among teenagers). (2) We found that this protocol produces enduring neurobehavioral alterations, most (but not all) of which appear to be rooted in persistent disruption of microglial homeostasis. (3) We uncovered unexpected modifications in the adult metabolic and immune phenotype of male and female ado-THC mice. (4) We published 29 articles in peer-reviewed journals, including Cell Metabolism, Biological Psychiatry, Nature Communications, and Nature Neuroscience. Service: we (1) organized 2 international symposia (>1600 participants), 5 research seminars, and 4 workshops/webinars; (2) created and launched a public biobank, which in ~12 months of service distributed >500 ado-THC tissue samples to 6 laboratories worldwide; (3) trained 9 graduate students (5 from underrepresented minorities, URM), 5 post-docs (2 from URM), and 51 undergraduate students (26 from URM); and (4) funded 7 pilot research projects. In this renewal, we propose to test the highly innovative hypothesis that ado-THC alters ECB signaling in microglia and, by doing so, disrupts both their homeostasis and their interactions with neurons, ultimately causing persistent (but possibly correctable) alterations in neuroplasticity, episodic memory, and vulnerability to opioids. This hypothesis, which is supported by rigorous preliminary data generated by the close cooperation of ICAL’s three Projects, will be tested in Aim 1 using emerging molecular techniques (e.g., single-nuclei RNAseq, imaging mass cytometry) along with state-of- the-art electrophysiological and behavioral methods. In Aim 2, we will continue our service mission by (1) organizing symposia and workshops to strengthen the cannabinoid research community and increase public access to cannabis-related information; (2) expanding the public reach of ICAL’s biobank; and (3) attracting early-career scientists to cannabinoid research by offering training opportunities and pilot funds.

NIH Research Projects · FY 2025 · 2018-05

Project Summary Research and training in organic synthesis impacts all fields of science that require the design and construction of molecular architecture. This project will advance catalytic methods, including hydroacylation and hydroamination, as attractive and powerful tools for chemical synthesis. The coupling methods proposed are modern and innovative because they rely on common functional groups (e.g., aldehydes, amines) to generate new carbon-carbon and carbon-nitrogen bonds from unsaturated partners (e.g., styrenes, dienes, cyclopropenes), with high selectivity and atom economy. Through experimental and theoretical studies, this project will yield fundamental insights into the mechanism of various transition metal catalysts, including rhodium, cobalt, and copper. Beyond catalysis and mechanistic studies, we plan to build chemical motifs of high significance to the field of drug discovery. We target privileged motifs (e.g., chiral nitrogen-containing heterocycles) and explore chemical space (e.g., tricyclic cages bearing high sp3 character). Through partnerships with experts in other fields, we address exciting challenges in fluorescent microscopy, NMR spectroscopy, and cancer immunotherapy by designing functional molecules. This project demonstrates innovation in making molecules at the interface of catalysis, medicinal chemistry, and biology.

NIH Research Projects · FY 2026 · 2018-04

Many insects pose major health and economic hazards to humans as common disease vectors and agricultural pests. Almost all of our present understanding of insect phototransduction is based on opsin-based photoreception in eyes that mediate image forming vision. My lab has recently discovered two additional phototransduction mechanisms in Drosophila. Cryptochrome (CRY) and Rhodopsin 7 (Rh7) expressed in central brain neurons mediate rapid onset sustained electrophysiological responses in these neurons. CRY and Rh7 light signaling underlie a novel form of non-image forming vision that strongly modulates complex time-of-day dependent insect behavioral responses to light, including avoidance/attraction behavioral choice between light and shade and light evoked arousal. While CRY's mechanism of action is due to light evoked redox state changes of its flavin adenine dinucleotide (FAD) chromophore and Rh7's mechanism of action is through a G- protein signaling pathway, they physiologically interact and may form the basis of a true color vision system for non-image forming vision that discerns specific light spectra. We have extended our study of non-image forming vision to harmful nocturnal Anopheles gambiae and diurnal Aedes aegypti mosquitoes and find that CRY1s mediate very distinct time-of-day dependent species specific behavioral light responses in these mosquitp-o important disease vectors. Remarkably, nocturnal and diurnal mosquito CRY1s confer mosquito species specific behavioral effects when expressed in all CRY expressing cells in a cry-null Drosophila genetic background and nocturnal mosquito CRY1 is significantly more light sensitive than diurnal mosquito CRY1 measured by multiple behavioral and electrophysiological assays. We will determine the detailed mechanisms that confer species specific physiological and behavioral light responses for flies and mosquitoes and other insects using a highly sensitive electrophysiological assay that we have developed that will allow us to accurately measure redox state changes and biological outputs for light sensitive CRYs and functional interactions between CRYs and Rh7, in combination with behavioral analysis. Our custom designed instrumentation allows us to examine CRY spectrally driven redox state changes in vivo. Present insect control strategies rely heavily on highly toxic pesticides. A far more environmentally friendly alternative is to make use of light-based behavioral manipulation of insects in a species specific fashion to attract harmful insect species to traps or to repel them away from human habitation. The goal of our research to form a rational basis for designing innovative new LED devices for species-specific harmful insect control in the ongoing fight against vector-borne diseases.

NIH Research Projects · FY 2026 · 2018-02

PROJECT SUMMARY The Hippo pathway is a key regulator of development, regeneration, tissue homeostasis and organ size, whose dysregulations have been frequently observed in human diseases like cancer. In mammals, the Hippo pathway is composed of a core kinase cascade that comprises two Ser/Thr kinases MST and LATS as well as their adaptor proteins SAV1 and MOB1, respectively, downstream effector protein YAP, and nuclear transcription factor TEAD. Upon Hippo pathway activation, MST phosphorylates and activates LATS, which in turn phosphorylates YAP, resulting in its cytoplasmic retention and degradation. Un-phosphorylated YAP is translocated into the nucleus, where it binds TEAD to drive the transcription of genes involved in various growth-related events. Recent studies in both Drosophila and mammals have further revealed MAP4K-family kinases (MAP4Ks) and PP2A phosphatase complex STRIPAK as additional components of the Hippo pathway, where MAP4Ks act in parallel to MST to phosphorylate and activate LATS, while STRIPAK inhibits MST and MAP4Ks to control the Hippo core kinase cascade. Multiple growth-related signaling events, such as growth factors, glucose/energy homeostasis, cell-cell contact, mechanical cues and a series of stress signals, have been uncovered to regulate the Hippo pathway and its downstream YAP-dependent transcriptional program. However, precisely how these upstream signaling stimuli feed into the Hippo pathway core components has not been fully understood. In the past grant period, we shed light on this long-standing question by characterizing phosphatidic acid and its-associated lipid metabolic pathway in transducing the Hippo pathway upstream signaling events to the Hippo pathway kinase LATS. In the current grant period, we propose to re-examine the Hippo pathway in energy stress response that was discovered by us a few years ago. Specifically, we will elucidate the molecular mechanisms underlying the energy stress-induced Hippo pathway activation by characterizing the role of the AMPK-STRIPAK-MAP4Ks axis in this process (Aim 1). In addition, we will investigate a YAP-independent function of the Hippo pathway in promoting cell survival against energy stress (Aim 2). Collectively, completion of this project will reveal mechanistically how energy stress activates the Hippo pathway and provide functional insights into the Hippo pathway in growth control and cancer development.

NIH Research Projects · FY 2026 · 2017-09

Project Summary/Abstract Our proposed project is to test the hypothesis that calcineurin (CN) inhibition may be a promising intervention to prevent or slow Alzheimer disease (AD). The molecular target of our treatment strategy, CN, has emerged as a key mechanism related to AD pathophysiology. Signs of CN hyperactivity are found during early stages of cognitive decline in humans and in mouse models of AD. Studies across numerous laboratories, using a variety of experimental models, suggest that CN activity is both necessary and sufficient for the progression of key AD markers including Aβ deposition, neurodegeneration, neuroinflammation/glial activation, synapse dysfunction, and cognitive loss. To inhibit CN, we will use two treatments: 1) tacrolimus, an FDA-approved drug used for the prophylaxis of allograft rejection and a second line treatment for numerous immune/inflammatory disorders and; 2) Q134R, a novel hydroxyquinoline derivative that inhibits the CN-dependent transcription factor, NFAT (but does not inhibit CN activity). In rodent models, tacrolimus and Q134R exhibit anti-inflammatory and neuroprotective properties. Moreover, an epidemiological study found that the incidence of dementia was strikingly reduced in human kidney transplant patients administered tacrolimus, relative to age-matched subjects in the general population. We are using the preclinical canine model of human aging and AD. Beagles are metabolically similar to humans and spontaneously develop amyloid-β (Aβ) deposition and cognitive decline with advanced age. Further, the aging beagle shows predictive validity in regard to several high-profile anti-AD drug trials. In this project, we proposed to extend an ongoing longitudinal prevention study, initiated in middle aged 5-8 year old beagles that are being treated with tacrolimus, Q134R or placebo. At the age we initiated the intervention, most animals were cognitively intact and expected to have little or no brain Aβ. Dogs have been treated for 2.5 years and will complete 3 years of treatment prior to this new study where we propose to extend the treatment study to 5 years in total. One group of 15 dogs is being treated with tacrolimus (0.075 mg/kg/day, orally), a second group of 14 dogs is receiving Q134R (8 mg/day orally) and one group is serving as a placebo control group (n=14). Aim 1 will continue to assess multiple longitudinal cognitive outcomes including learning, executive function, spatial and object recognition memory. Aim 2 will expand on measures of plasma and CSF levels of AD biomarkers (e.g. NfL, Aβ, GFAP). Aim 3 will continue MRI measures of structure, and metabolic and vascular pathology to detect in vivo outcomes reflecting brain health. Aim 4 will focus on neuropathology (Aβ, glial activation, synapse dysfunction, neurodegeneration) and CN related pathway modifications. All outcome measures in this study are similar if not identical to those used in human clinical trials (including a human MR scanner, fluid biomarkers, assessment of analogous cognitive domains). We hypothesize that tacrolimus and Q134R will lead to maintenance of cognition, maintenance of CSF and plasma biomarker outcomes reflecting reduced brain pathology, maintenance of structural integrity, metabolic function and reduced vascular pathology and reduced AD neuropathology. These studies will provide a rigorous test of the CN hypothesis of AD and possibly pave the way for investigating if CN inhibition may serve as a primary or complementary treatment strategy in human AD clinical trials.

NIH Research Projects · FY 2025 · 2017-09

PROJECT SUMMARY Poverty puts children at risk for developmental delays, lower school achievement and educational attainment, and unfavorable labor market and health outcomes. The Baby’s First Years (BFY) project is the first large-scale randomized controlled trial in the U.S. to estimate the impact of poverty reduction on children’s development and health. Launched in 2018 (NICHD R01HD087384), BFY recruited 1,000 low-income mothers and their newborn infants in four metropolitan areas. Mothers were randomized to receive a monthly unconditional cash transfer of either $333 (“high-cash gift group”) or $20 (“low-cash gift group”) for the first 4 years and 4 months (52 months) of the child’s life. Participants have been followed up annually around the children’s birthdays to measure child development and family life. In this renewal, with funding already in hand for a two-year extension of the cash gifts, BFY has the opportunity to study the impact of poverty reduction for an unparalleled duration, across the first six years of life. The continuation of the project is driven by a need to understand whether continuous monthly cash transfers will improve low-income children’s development at the start of formal schooling. To accomplish this, we will collect two lab-based waves of data, at ages 6 and 8. We will assess high-cash/low-cash group differences at age 6 on measures of academic achievement skills as well as cognitive, self-regulation, and socio-emotional development. We will additionally assess high-cash/low-cash group differences in measures of brain activity and stress physiology. At age 8, we will investigate whether children’s learning and developmental trajectories have been altered in ways that generate persistent impacts, 20 months after the cessation of the payments. The study will also measure family contexts based on two pathways by which poverty is theorized to affect children: an investment pathway (household expenditures; maternal work; activities with children, early care and education arrangements) and a stress pathway (economic hardship; parental relationship quality, maternal mental health, stress, and parenting quality). At both ages 6 and 8, we will assess high-cash/low-cash group differences in these investment and stress pathways.

NIH Research Projects · FY 2025 · 2017-09

ABSTRACT The goal of the University of California, Irvine MODEL-AD U54 Center is to develop novel mouse models of late- onset Alzheimer’s disease (LOAD), to deeply phenotype these and to make all data and mouse strains available to enable researchers to select the optimal mouse model and timepoints for therapeutic and intervention testing, as well as testing of hypotheses concerning mechanisms of LOAD. During the past five years, we have generated and deeply phenotyped mice with one component of our base genetic platform in which the Aß region of the App gene was converted from the rodent to the human sequence, and we have recently introduced the second component, a humanized MAPT (TAU) locus produced via gene-replacement. We have also used CRISPR and genome replacement to model and validate nine GWAS identified LOAD risk loci and have characterized mice with each of these both on a wild-type and 5xFAD background to determine their effects on plaque generation and damage exerted on the brain in response to pathology. In this continuation, we will use the results of these analyses to identify the combinations of LOAD risk variants most likely to phenocopy LOAD and introduce them on two complementary hAb-KI, hTAU, hAPOE4 platform lines, designed to mimic sub-types of AD that have been recently defined. To ensure translationability, we have an expanded focus on biomarkers and alignment with human phenotypes. To that end we have established a new Core – the Neuroimaging and neurovascular core (NIVC), which will provide brain imaging modalities currently employed in human AD subjects to align phenotypes in our mice with human disease progression. We have also expanded our fluid biomarker analysis efforts to include CSF, as well as plasma lipidomics and metabolomics to be compared to human AD plasma signatures. Similarly, our bioinformatics and data management efforts have been expanded to include single cell and nucleus RNA-seq and ATAC-seq, as well as spatial transcriptomics to enable alignment of data from our models with human AD signatures, but also to understand the mechanisms underlying disease progression in our mice. We are utilizing a comprehensive approach to evaluate our mice across their lifespans, which includes behavioral/cognitive assessment, electrophysiological analysis, super-resolution synaptic imaging, neuroimaging, bulk and single-cell RNA-seq, single cell level spatial transcriptomic analysis, unbiased proteomics, and microbiome and metabolome investigations. The UCI MODEL-AD Center will leverage the resources of our NIA-funded Alzheimer’s Disease Research Center combined with AMP-AD and other human AD datasets to facilitate comparisons to the human condition to identify the best mouse models to evaluate further. All data and models will be made available without restrictions, via The Jackson Labs, and data will be explorable via the modeladexplorer.org website, and raw data freely available for download via the AD Knowledge Portal.

NIH Research Projects · FY 2026 · 2017-09

Abstract Facioscapulohumeral dystrophy (FSHD) is one of the most common muscular dystrophies in the U.S. Currently, there is no effective treatment, and the pathogenic process is still not completely understood. Most cases (>95%) of FSHD involve mono-allelic deletion of macrosatellite D4Z4 repeat sequences at the subtelomeric region of chromosome 4q (FSHD1), while the remaining ~5% of cases demonstrate no D4Z4 repeat contraction (FSHD2). Mutations in the SMCHD1 genes were linked to FSHD2, and also greatly exacerbate the phenotype of FSHD1 by acting as a modifier of the disease severity. Expression of the DUX4 gene encoded within the D4Z4 repeat is critically linked to the development of FSHD. Since overexpression of DUX4 is cytotoxic in human myocytes and mice, it is thought that DUX4-induced cytotoxicity is the cause of dystrophy. However, only ~0.1% of patient muscle cells appear to express DUX4, and DUX4 expression can occasionally be observed in muscle cells from unaffected individuals. It is not straightforward to study FSHD in model organisms as D4Z4 repeats and some of the critical DUX4 target genes are primate-specific. During the previous funding period, we found evidence that DUX4-negative patient myocytes exhibit altered gene expression distinct from control myocytes and cross-regulation of DUX4 target transcription factors contributing to sustaining the DUX4 gene network. These findings strongly argue that FSHD mechanism is not simply DUX4-induced cell killing and further investigation is necessary to understand FSHD pathogenesis. We also developed genetically engineered mutant myoblast lines carrying D4Z4 deletion, SMCHD1 mutation or both to simulate FSHD1, FSHD2 and severe cases of FSHD1, respectively and began to characterize epigenetic and gene expression consequences of defined mutations in the isogenic background. Specific Aims of this project are (1) to create additional mutant clones to interrogate the consequences of FSHD mutations in different muscles with different disease susceptibility and during early myogenesis; (2) to investigate dynamics and regulation of DUX4 and target gene expression, and (3) to identify a modifier gene(s) that dictates disease susceptibility and severity. The successful outcome of the project should reveal DUX4 and target gene dynamics and their contributions to FSHD pathogenesis and identify critical determinants for the disease susceptibility, which may lead to identification of potentially new therapeutic targets.

NIH Research Projects · FY 2026 · 2017-08

Circadian rhythms are fundamental for understanding biology: they date to the origin of life, are found in virtually every species from cyanobacteria to mammals, and coordinate many important biological functions from the sleep-wake cycle, to metabolism, to cognitive functions. Circadian rhythms are equally fundamental for health and medicine: diet modifications have been linked to molecular-level changes in circadian rhythms; disruptions of circadian rhythms have been linked to health problems ranging from depression to learning disorders to diabetes, to obesity, to cardiovascular disease, to cancer, and to premature aging; finally, a large fraction of drug targets have been found to oscillate in a circadian manner in one or several tissues. A better understanding of circadian oscillations at the molecular level has many direct applications to precision health and medicine. To illuminate circadian oscillations at the molecular level, modern high-throughput technologies are being used to measure the concentrations of many molecular species, including transcripts, proteins, and metabolites along the circadian cycle in different organs and tissues, and under different conditions. Yet informatics tools for processing, analyzing, and integrating the growing wealth of molecular circadian data are not yet in place. This effort will fill this fundamental gap by continuing to develop and disseminate informatics tools to enable the collection, integration, and analyses of this wealth of information and lead to novel and fundamental insights about circadian oscillations' organization and regulation, roles in health and disease, and future applications to precision medicine. Specifically, via close collaborations among computational and experimental scientists, this effort will have four main aims: (1) Data: Aggregate the largest possible collection of circadian omic (e.g., transcriptomic, metabolomic, proteomic) experimental datasets covering as many species, cells, tissues, organs, and conditions (e.g., genetic, epigenetic, environmental) as possible. (2) Analysis: Develop analytical tools, including deep learning tools, to analyze these datasets to identify molecular species with a periodic concentration profile with statistical determination and conduct integrated differential analyses across the different datasets. (3) Web Platform: Import the analyses' datasets and results into an integrated database and serve them publicly through a web server (CircadiOmics platform) as a one-stop shop for viewing or downloading circadian data, annotations, tools, and analyses, enabling other scientists to view and analyze circadian data comparatively. And (4) Applications: Apply the CircadiOmics platform's datasets and tools to specific biomedical problems via multiple efforts in collaboration with other experimental labs to identify the role of circadian oscillations in health and disease and generate mechanistic molecular hypotheses that can then be tested in the lab. One example of such collaboration is the study of the interplay between Alzheimer's disease and circadian rhythms. All data, software, and results will be freely available for academic research purposes and broadly disseminated through multiple channels to benefit the biomedical community and society at large.

NIH Research Projects · FY 2026 · 2017-08

PROJECT SUMMARY A critical gap in understanding the etiology of sporadic Alzheimer's disease (AD) is identifying the upstream factors that lead to the development of both Alzheimer’s pathology and related neural dysfunction. Vascular disease is found in approximately 80% of patients with concomitant AD pathology and thus may be an important contributor to the development of AD, however relationships between vascular health and the emergence of AD pathophysiology has not yet been comprehensively investigated in cognitively normal samples. While large vascular adverse events such as stroke are known to confer risk for developing vascular dementia, growing evidence suggests that subtle vascular damage accrued through a lifetime of injury could predispose neural structure and function to become more susceptible to AD-related pathophysiology. Critically, chronic and subtle forms of vascular disease are more commonly found in Black and Hispanic populations with reduced access to healthcare and could help explain the increased prevalence of AD in these populations. The goal of this renewal project is to establish the role of cerebrovascular injury and dysfunction (CVID) in the pathophysiology of preclinical AD and develop individualized imaging-based cerebrovascular profiles that predict memory decline across racially and ethnically diverse populations. We will conduct follow-up assessments in 100 nondemented older adults (over 60 years of age) in our current award (BEACoN Cohort: R01AG053555), which includes amyloid-PET (florbetapir), serial high-resolution MRI and tau-PET (MK-6240), our innovative digital cognitive biomarkers which assess pattern separation, and a full UDS-3 neuropsychological testing battery. We will complement this with targeted new recruitment (n = 100) to increase the representation of Hispanic/Latino and Black participants in our cohort. We have built an infrastructure to radically transform recruitment and retention in our study including innovative partnerships with clinical research organizations with a demonstrable track record in minority recruitment. Given focus on subtle vascular damage, we will exclude based on history of stroke or severe cardiovascular disease. Our aims are (1) Assess the novel biomarker framework in which CVID predicts tau accumulation, which predicts structural and functional deterioration of the medial temporal lobes (MTL), subsequently predicting decline in hippocampal pattern separation. (2) Construct individualized brain imaging based CVID profiles that differentially predict decline in hippocampal memory across racially and ethnically diverse populations. (3) Aim 3: Associate CVID profiles with modifiable lifestyle risk factors and structural and social determinants of health that are differentially distributed across racial and ethnic groups. In summary, we will develop a novel mechanistic framework for how CVID contributes to AD pathophysiology and memory/cognitive decline that directly addresses racial and ethnic disparities in AD risk. Cerebrovascular profiles, and their associated modifiable risk factors that confer the greatest risk of AD, will be identified as targets for future intervention.

NIH Research Projects · FY 2024 · 2017-06

This research will develop methods to model active sites in metalloproteins for the purpose of determining fundamental structure-function relationships for how proteins activate dioxygen, a process that strongly impacts human health and aging. Artificial metallproteins will be prepared utilizing biotin-streptavidin technology as a tool to ensure specific and reproducible placement of synthetic metal complexes within protein hosts. This approach is proposed to be an effective method to model key properties of the active sites in native metalloproteins, including site isolation of species, regulation of the primary coordination sphere, and control of the microenvironments around the metal complexes. One glaring weakness of many biomimetic systems is their limited ability to regulate the microenvironments that surround metal centers. No chemical system operates in isolation without interacting with its local environment. There is a growing body of evidence from structural biology that the microenvironment, a space around metal complexes that comprises the secondary coordination sphere, has profound effects on protein function that ranges from modulation of physical properties to delivery of reactants and removal of products. It is our contention that the greater regulation of microenvironments will lead to better understanding of protein function. It is further maintained that the benefits gained from fundamental analyses as proposed in this application extend well beyond improvements in selectivities/efficiencies at the molecular level – they are transformative for all types of platforms, providing the requisite information that is still missing for the development of highly functional systems. We propose an approach for preparing artificial metalloproteins that allows for the confinement of synthetic complexes within protein hosts to regulate both the primary and secondary coordination spheres about the immobilized metal centers. The ability to regulate these coordination spheres within a protein will produce systematic structure-function relationships that will lead to an improved understanding of chemical processes that are directly linked to human health.

NIH Research Projects · FY 2026 · 2017-04

Malaria Epidemiology and Vector Biology of Invasive Anopheles stephensi Across Rural and Urban Landscapes in Ethiopia PROGRAM SUMMARY Anopheles stephensi is a major malaria vector species in South Asia. Since its first detection on the African continent in Djibouti in 2012, the distribution of this vector species has expanded to Ethiopia, Sudan, Somalia, Kenya, and Nigeria. The emergence and spread of An. stephensi in Africa pose serious challenges for malaria control and elimination in fast-growing urban Africa. Knowledge gaps related to An. stephensi ecology and behavior and the effectiveness of intervention methods in Africa have impeded the development of effective malaria control programs. Lack of sensitive surveillance methods for An. stephensi has also hindered efforts to effectively track An. stephensi population spread over time. To date, only limited basic research has examined the biology of invasive An. stephensi mosquitoes and the impact of An. stephensi invasion on malaria epidemiology in Africa. Little translational research has been conducted to develop new surveillance and control tools for An. stephensi. To address the major knowledge gaps and challenges in malaria control and elimination in the face of An. stephensi invasion and rapid spread across sub-Saharan Africa, our ICEMR established a consortium of outstanding institutions and investigators from the U.S. and Ethiopia to study critical scientific questions regarding malaria control and elimination efforts in the Greater Horn of Africa. The overarching goals of this ICEMR are to 1) address the knowledge gaps in invasive An. stephensi vector biology and malaria epidemiology across the rural to urban continuum in Ethiopia; 2) develop surveillance tools and methods needed to track vector spread; and 3) identify cost-effective vector control methods that can be adapted to settings of varying malaria endemicity. The ICEMR has two projects, each with multiple specific objectives. Project 1 will assess the vector biology of An. stephensi, elucidate the extent of spread of invasive An. stephensi, and develop serological biomarkers for An. stephensi exposure surveillance. Project 2 will examine the impact of An. stephensi invasion on malaria risk and determine the epidemiological impact and cost-effectiveness of larviciding methods targeting areas of high malaria risk across the rural to urban landscape. The administrative and data management cores will provide support to the entire program. Knowledge gained through this ICEMR is important to malaria control and elimination, not only in Ethiopia, but also in other African countries with invasive An. stephensi.

NIH Research Projects · FY 2024 · 2016-12

PROJECT SUMMARY The Molecular Transducers of Physical Activity Consortium (MoTrPAC) is designed to discover and characterize the range of molecular transducers that underlie the effects of exercise in humans. MoTrPAC was launched in 2016 with six adult clinical centers and a pediatric center that have collaborated to generate extensive Manual of Operations to guide research protocols involving all aspects of the clinical operations (Phase I). Phase II began in the fall of 2019 with all human clinical centers showing excellent progress towards initial recruitment goals and implementation of the protocol. The initial goal set forth by NIH was to recruit 270 children (10-17 years of age) and 1980 sedentary adults (age 18 years or greater) that are randomized to endurance training (170 youth, 840 adults), resistance training (840 adults), or non-exercise controls (50 youth, 300 adults). An additional group of highly active endurance (50 youth, 150 adults) and resistance (150 adults) trained individuals serve as comparators and are not participating in the MoTrPAC exercise training programs. The recruitment and enrollment approach are sex balanced and with participants across a wide range of ages (10-17, 18-39, 40-59 and >60-year age groups) and of different races. Due to the COVID-19 pandemic, MoTrPAC activities were suspended for over a year (beginning in March 2020) with continued constraints through 2022. Despite the numerous challenges encountered as a result of the pandemic, recruitment activities at the adult and pediatric clinical centers have accelerated to a rate that is projected to successfully achieve the target enrollment numbers by the end of the new award period. This led to the NIH Common Fund to release the current NOFO (RFA-RM- 23-010) to provide MoTrPAC with funding to complete recruitment and follow-up for the clinical studies, including finishing mechanistic randomized controlled trials of sedentary adults and children and observational studies of highly active adults and children. This will enrich the participant cohorts that are critical to understand exercise adaptations and heterogeneity across age, gender, and minority groups. Altogether, this extension will allow MoTrPAC to complete the intended goals as originally envisioned and will provide a more complete public database of the health benefits of exercise and provide insight into how physical activity mitigates disease.

NIH Research Projects · FY 2025 · 2016-09

Ribosomes are responsible for protein synthesis in the cell and are essential for growth and cell division. Genetic defects that interfere with ribosome biogenesis reduce translation and growth and cause human diseases. Heterozygous mutation of many ribosomal protein genes causes Diamond-Blackfan Anemia, which is associated with early onset anemia, morphological defects, and cancer predisposition. Ribosomal protein genes are also haploinsufficient in the fruitfly Drosophila, where they cause reduced protein synthesis, slow growth and development, and morphological defects. Unexpectedly, these effects are found to have a transcriptional basis, that is, ribosomal protein mutations activate a transcriptional program which controls translation and the other effects. This project will determine how ribosome biogenesis defects cause human diseases and cancer by elucidating the molecular mechanisms of this transcriptional response in Drosophila, and exploiting mutations that prevent cells reacting to ribosomal protein mutations. The project will use ribosome profiling methods to define the hierarchy of steps at which translation of the genome into protein is altered and highlight those likely to be relevant to cancer, anemia, and morphology. The project will use gene modification approaches to define the specific roles of individual protein isoforms and domains in the effects of ribosome biogenesis defects. The project will explore what physiological and selective role is played by the cellular responses to ribosome biogenesis defects, and their contribution to promoting health and survival and preventing disease. The findings are anticipated to suggest approaches to prevent cancer and treat the ribosomopathy Diamond Blackfan Anemia.

NIH Research Projects · FY 2026 · 2016-08

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer mortality in the US. Surgical resection is the only curative treatment, and due to the high recurrence rate (60%), only 20% survive five years. Chemotherapy and radiotherapy after surgical resection only offer modest improvements in survival. A new approach to prevent tumor recurrence in PDAC patients is urgently needed. We will compare overall survival (OS) after tumor resection between the combination of dendritic cell (DC) vaccination via an improved intraperitoneal, i.p., route, combined with current therapy (gemcitabine), versus the use of preventive DC vaccination alone or gemcitabine alone (Aim 1). Given the relationship between migration to lymph nodes (LNs) and anti-tumor immune response, accurate quantification of DC vaccine migration could serve as an early biomarker for predicting longitudinal response (OS) and elucidating the cause of differential response rates between patients. Understanding factors that affect DC vaccine response rates will enable the titration of vaccine doses to optimize outcomes for individual patients. Thus, we will validate magnetic imaging via quantitative susceptibility mapping (QSM) and ultrashort echo time (UTE) R2* techniques for tracking clinically applicable magnetic-labeled DC vaccines to draining abdominal LNs (Aim 2). We will test whether advanced MRI-tracked DC vaccine homing to LNs can be used as an early imaging biomarker to predict OS of the combination of DC vaccination and gemcitabine treatment post-surgery (Aim 3). The proposed work will meet the significant demand for a novel DC vaccination strategy of cancer therapy that can rapidly be translated to the clinic to prevent relapse after pancreatic tumor surgery while adding an imaging biomarker as a potentially powerful method to simultaneously predict response to the therapy. The success of the proposed preventive DC vaccination strategy and prediction of response to treatment could have a broad impact as a clinical extension to other solid organ systems (e.g., stomach, liver, colorectal, renal, or uterine tumors) as novel adjuvant immunotherapy to prevent relapse after surgery.

NIH Research Projects · FY 2025 · 2016-06

Program Director/Principal Investigator (Sevrioukova, Irina F.): Project Summary Human cytochrome P450 3A4 (CYP3A4) is the major and most clinically relevant drug-metabolizing enzyme, notoriously known for its extreme substrate promiscuity and allosteric behavior. Drugs and other xenobiotics can also stimulate and inhibit CYP3A4 activity, which frequently leads to undesired drug-drug interactions (DDIs), chemical toxicity and therapeutic failures. Despite extensive investigations, the CYP3A4 inhibitory and activation mechanisms remain incompletely understood. This proposal centers on using structural biology approaches to address key issues in both areas of CYP3A4 research. Aim 1 is set to investigate the CYP3A4 inhibitory mechanism via rational structure-based design of analogues of ritonavir, an HIV protease inhibitor whose ability to potently inhibit CYP3A4 was purely coincidental. We will identify structural determinants required for potent inhibition by rationally designing and investigating structure-activity relations of ritonavir-like compounds and, based on our findings, build a 3D-pharmacophore model for a potent CYP3A4-specific inhibitor that can be used for early prediction/elimination of the inhibitory potential in drug candidates and for development of more effective pharmacoenhancers. Aim 2 will utilize an integrated biochemical, chemical labeling, structural and computational approach to investigate the CYP3A4 substrate binding cooperativity and allosterism. Our recent structural findings confirmed the importance of the previously mapped peripheral area and identified three novel inner sites that could serve for substrate/effector docking. We will evaluate the role and relative importance of these areas by assessing how their modification/disruption affects CYP3A4 conformation, substrate binding cooperativity, stoichiometry and metabolism. The research outlined in this proposal is important from both the basic and translational science perspectives, because it will fill the knowledge gaps and provide fundamental insights into plasticity and adaptability of CYP3A4 to structurally diverse ligands, clarify molecular mechanisms underlying the complex ligand binding behavior and oxidative kinetics, and help develop better tools for in silico prediction of protein-ligand contacts, metabolic stability and DDI potential in drug candidates to improve their efficacy and reduce off-target effects. OMB No. 0925-0001/0002 (Rev. 08/12 Approved Through 8/31/2015) Page Continuation Format Page