Stanford University

NIH Research Projects · FY 2025 · 2024-07

Project Summary Radiation-Induced Cardiotoxicity (RIC) remains a concerning health issue, particularly in accidental radiation exposure scenarios as evident in the Life Span Study of Japanese atomic bomb survivors. However, the complex interplay of factors contributing to the diverse presentations of RIC remains elusive. This project aims to shed light on the critical roles of sex hormone “estrogen” and genetic variations in estrogen receptor (ER) signaling in modulating RIC susceptibility and response. The research takes an innovative approach by combining state-of- the-art techniques and multidisciplinary expertise. Firstly, a "cell village" strategy will be employed which leverage pooling human induced pluripotent stem cell (iPSC) derivatives from a diverse cohort of 200 individuals. Pooled iPSCs will be differentiated into 3D cardiac organoids (iPSC-COs) and treated with varying doses of estrogen and radiation to simulate different physiological conditions and radiation exposure, respectively. Cutting-edge single-cell genomics and computational technologies will be employed to scrutinize the resulting transcriptomic and epigenomic changes in each individual. This will help us identify inter-individual variations and underlying genetic mutations that contribute to differential molecular responses upon irradiation. Additionally, animal models will be employed to simulate radiological incidences and corroborate multi-omics data to functional outcomes in whole organisms. Collectively, these experiments will elucidate the genes responsible for sexual disparity in RIC and its relation to estrogen signaling which can provide insights into personalized risk prediction and intervention strategies, addressing a critical knowledge gap in the field of radiation biology and cardiovascular health.

NIH Research Projects · FY 2025 · 2024-07

Dilated cardiomyopathy (DCM)-associated heart failure is a leading cause of death. About a third of all DCM is caused by variants in genes controlling heart muscle structure and/or function. Although this information is improving patient management, it has not yet led to new therapeutics. This proposal is to evaluate the potential of activating the AMPK-BACH1-NRF2 signaling pathway to treat inherited forms of DCM. The pathway emerged from an unbiased, high-throughput functional genomics screen to discover novel therapeutic targets and signaling pathways capable of restoring contractile function in human induced pluripotent stem cell cardiomyocytes (hiPSC-CMs) harboring DCM-causing mutations. The screen and subsequent validation experiments revealed that activation of the AMPK-BACH1-NRF2 pathway restored contractility of multiple patients’ hiPSC-CMs carrying the DCM-causing TNNT2 R173W mutation. The magnitude of the effect was comparable to CRISPR-mediated correction of the mutation. Importantly, activation of the pathway had no effect on healthy (isogenic control) hiPSC-CMs, suggesting that the therapeutic response is specific for the disease context. Modulation of AMPK, BACH1 and NRF2 have been reported to elicit protective effects in ischemic heart disease models but these targets are largely unexplored in the context of inherited DCM. The proposed Specific Aims address critical issues necessary to advance targeting this pathway to treat inherited DCM. AIM 1 explores the selectivity of pathway activation for restoring contractile function in different forms of inherited DCM, including disease caused by mutations in myofilament (TNNT2, MYH7 and TTN) and non-myofilament (PLN, RBM20, LMNA) genes. We expect that the pathway will ameliorate contractile dysfunction in multiple forms of DCM. AIM 2 will define the mechanism(s) of action by identifying specific genes downstream of pathway activation that are essential to restore contractile function. Our hypothesis is that the pathway converges on a subset of NRF2- target genes. AIM 3 explores whether activating the pathway restores contractile function by normalizing metabolic and/or ER/SR stress. Finally, AIM 4 tests whether activating the pathway will mitigate clinical features of DCM using an established mouse model. A successful outcome of this project will constitute a mechanism-based approach to treat inherited DCM. The project uses both human patient iPSC models and mouse transgenic models to integrate human cellular context with whole organism physiology. The outcome will define mechanism(s) of action that might not only benefit inherited DCM but also inform the development of novel strategies to treat acquired forms of DCM.

NSF Awards · FY 2024 · 2024-07

Non-technical description Biologists have recently discovered how to grow patient cells into little spheres that resemble human tissue. These engineered tissues can be used to safely study how different drugs might interact with our cells, without exposing the patient to any drug treatments might be potentially harmful. Unfortunately, these new biological methods have three big challenges: they are not very reproducible, it is hard to control how the cells grow over time, and the spheres cannot be easily attached together to make larger tissues that more accurately model real patients. In this work, we develop new gel materials that will help to overcome all three of these challenges. Our gels are designed to resemble the materials that surround the cells in our body. These materials will provide biochemical signals to the cells and also provide physical support as the cells grow, divide and move around to form larger tissues. To demonstrate that our gels are reproducible and versatile, we will grow models of three different types of patient tissue: intestines, brain, and glioma (i.e. brain cancer). The research team will also lead an outreach program at a local elementary school to showcase how scientific exploration can be fun and exciting. The research will also include interns from local community colleges to provide hands-on scientific training. Technical description Organoids (i.e. stem cell-derived, multicellular structures with emergent self-organization into tissue-like structures) hold great potential for engineering 3D tissues in vitro, whether for the purposes of regenerative medicine, fundamental studies of human development, or personalized disease models. However, several key challenges currently hinder the formation of these complex, engineered tissues, including (i) reproducibility in organoid formation, (ii) controllable morphogenesis (i.e. organization of cell types within the organoid), and (iii) spatial control over multi-organoid patterning. Here, we take a biomaterials approach to address these three challenges by designing a new class of biopolymeric gels. We call this new family of biomaterials liposome network crosslinked (LINC) hydrogels. LINC hydrogels are networks of biocompatible polymers crosslinked through self-assembled lipid vesicles, or liposomes. Within each liposome, the individual lipids are in constant motion, and their motion can be controlled through tuning lipid design parameters. In Aim 1, we will design and characterize LINC hydrogels with tunable biochemical and mechanical properties to demonstrate modularity. In Aim 2, we will customize LINC hydrogels for the production of three different types of organoid building blocks. In Aim 3, we will customize LINC hydrogels for the bioprinting of organoid building blocks into complex tissue structures called assembloids. To demonstrate versatility of our biomaterials, we will focus on three biological case studies: human intestinal organoids from intestinal stem cells, human neural organoids from induced pluripotent stem cells, and pediatric glioma organoids from patients, as these are prototypical cultures for the three key application areas of organoids: regenerative medicine, models of human development, and precision medicine, respectively. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT. Ferroptosis is a form of regulated cell death defined by the iron-dependent accumulation of membrane lipid peroxides that results in oxidative damage and plasma membrane disruption. Ferroptosis is a form of cell death that is mechanistically and morphologically distinct from other forms of cell death, such as necroptosis, apoptosis, and pyroptosis. Targeting ferroptosis has garnered immense interest in the cancer research community owing to the potential to selectively active this mechanism of cell death in cancer cells. To realize the full potential of inducing ferroptosis in cancer, non-invasive imaging markers are needed. System xc-, the cystine-glutamate antiporter, is a plasma membrane transporter mediating the cellular uptake of cystine in exchange for intracellular glutamate. System xc- plays a critical role in ferroptosis by regulating the availability of cystine, a crucial precursor for glutathione biosynthesis, thus impacting the cellular antioxidant defense system and ultimately influencing susceptibility to ferroptosis. Modulation of system xc- activity with pharmacological inhibitors is a potential therapeutic strategy that regulates this form of cell death in anti-cancer therapy. Molecular imaging of system xc- therefore has the potential to identify cancers with appropriate transporter activity that would be suitable for ferroptosis induction, assess drug engagement and inhibition of system xc- in living subjects, and monitor the efficacy of targeting this emerging mechanism of cell death in cancer. To the best of our knowledge, the investigation of system xc- radiopharmaceuticals in this context of ferroptosis, has yet to be reported. We have pioneered the development of [18F]hGTS13, a homo-glutamate radiotracer specific for system xc-. Importantly, [18F]hGTS13 demonstrates several advantages over existing system xc- radiotracers including: 1) incorporation of a UV-active group, thus greatly facilitating radiosynthesis and quality control; 2) improved transporter specificity; and 3) reduced uptake in multiple immune cell types and improved cancer specificity. The goals of this proposal are to establish [18F]hGTS13 PET as a non-invasive imaging marker of drug induced cancer ferroptosis in cell culture and pre-clinical models and to clinically translate [18F]hGTS13 by determining its pharmacokinetics in healthy human volunteers through an exploratory investigational new drug mechanism. We will first establish the relationship between [18F]hGTS13 uptake and sensitivity to cancer ferroptosis in the in vitro and in vivo setting (SA1). We will then test the ability of [18F]hGTS13 to monitor the engagement of pro- ferroptotic drugs in vivo and predict treatment response (SA2). Lastly, we will translate [18F]hGTS13 for first-in- human testing and determine its biodistribution and radiation dosimetry in healthy volunteers (SA3). Successful completion of this project will establish the foundation for employing [18F]hGTS13 to select patients that are likely to be sensitive to ferroptosis inducing therapies and monitor treatment effects. The findings from this work will have high impact as inducers of ferroptosis advance to clinical investigation.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Interventions are critically needed to address the maternal mental health crisis in the United States, particularly since mental illness is now the leading cause of maternal mortality. Unprecedented societal shifts, disparities in access to mental health care, stigma, reluctance to use psychiatric medications, and inflammation associated with uncontrolled mental illness all highlight the need for simple, scalable, and equitable solutions in pregnancy. Developing solutions tailored to specific aspects of this problem is an essential first step. However, there is a significant knowledge gap in how mental illness contributes to maternal morbidity and mortality which must first be understood to direct risk reduction strategies. One approach to doing this involves studying severe maternal morbidity (SMM), a composite of severe complications that often precede death yet occur 70 times more frequently. In preliminary work, Dr. Panelli found that the risk of SMM is 58% higher in people with mental illness yet the relative contribution of other factors such as substance use and medical comorbidities to this risk remains to be elucidated. To address these knowledge gaps and prepare for a career as an independent clinician scientist in this field, Dr. Panelli has developed this innovative K23 proposal to reduce SMM due to mental illness through identifiable behavioral and biologic pathways. The Specific Aims are to: 1) Identify casual pathways for increased SMM in people with depression and anxiety – the two most common perinatal mental illnesses – using a population-level approach with Veteran’s Health Administration (VA) data; and 2) Conduct a pilot randomized controlled trial (RCT) of a physical activity intervention in pregnant people with mental illness to reduce severity of depression and/or anxiety and lower stress biomarkers (leukocyte telomere length and hair cortisol). These aims will be conducted in parallel to provide complimentary preliminary data to inform an R01 RCT of a physical activity intervention to reduce SMM in people with mental illness. Dr. Panelli’s training will be conducted under the experienced mentorship of Drs. Carmichael (perinatal epidemiologist), Phibbs (health economist), and Gotlib (clinical psychologist/neuroscientist), in conjunction with advisors in lifestyle interventions (Dr. Pinto) and RCT design (Dr. Lyell). Support from this K23, which will feature content training in perinatal mental illness, hands-on analysis of population-level data with advanced epidemiologic methods, and experience with clinical trials, will advance Dr. Panelli’s long-term goal of becoming an independent clinical researcher with expertise in the complex physical and biological interrelationship between mental illness, pregnancy, and SMM.

NIH Research Projects · FY 2026 · 2024-07

Building on the extensive genome wide association studies (GWAS) of coronary artery disease (CAD), and single cell characterization of atherosclerosis, we have shown that the smooth muscle cell (SMC) lineage harbors much of the risk for vascular disease. Further, these data indicate that SMC can assume two disease- related transition states linked to disease risk. We identified TCF21 as a protective CAD GWAS gene and showed that it regulates a disease-related transition of medial SMC to a fibroblast-like phenotype, producing cells we term “fibromyocytes” (FMC). Further, we and others have found that SMC can also transition to a second phenotype, characterized by expression of genes known for their role in endochondral bone formation and intimal vascular calcification. We showed in mouse genetic models that this chondrogenic process, which gives rise to cells we term “chondromyocytes” (CMC) is actively inhibited by the CAD GWAS protective Tgfb1 signaling molecules Smad3 and Zeb2, and promoted by the CAD GWAS causal factors Pdgfd and Twist1. Further, recent transcriptomic and epigenomic (multi-omic) single cell studies indicate that FMC and CMC are the end products of disparate trajectories that SMC differentially traverse in the disease setting. Our central hypothesis for this work postulates that these trajectories are specified by enhancers and key regulatory transcription factors (TFs) that mediate the branch point directionality of transition commitment, and that these cis- and trans-acting genetic modulators underlie the fundamental mechanisms of CAD gene causality in the SMC lineage. Algorithms that link disease variation and disease scRNAseq data show that disease genetic risk resides in cell state changes from SMC to FMC and SMC to CMC, making it imperative that we investigate the enhancers, related TFs, and regulons that mediate these phenotype transitions. Specifically, in Aim 1 we will generate and analyze a time-course of single cell multi- omic data from a mouse atherosclerosis model to map the epigenomic and transcriptomic regulatory elements that mediate SMC transition to the FMC and CMC trajectories. We will perform similar studies with mice targeted for CAD associated genes that promote or inhibit disease risk, to identify differences between the cis- and trans-acting factors, and disease-related regulons that determine the disease trajectories and directionality of disease risk. In Aim 2 we will perform multi-omic assays on human coronary lesions of varying severity to examine the molecular mechanisms that mediate SMC transitions in human disease. Finally, in Aim 3, we will perform in vitro enhancer PerturbSeq in human coronary artery smooth muscle cells to validate computational results from previous Aims, and further characterize downstream targets and gene regulatory networks of transition enhancers and TFs. The proposed studies will identify cellular and molecular mechanisms that mediate SMC transitions to FMC and CMC, and the relationship of these transitions to vascular calcification and disease risk.

NSF Awards · FY 2024 · 2024-07

Formal Methods in Computer-Aided Design (FMCAD) 2024 is the twenty-fourth in a series of conferences on the theory and applications of formal methods in hardware and system verification. It provides a leading forum for researchers in academia and industry to present and discuss ground-breaking methods, technologies, theoretical results, and tools for reasoning formally about computing systems. This grant will help support conference travel for up to 8 students enrolled in US institutions to attend FMCAD, which will be in Prague, Czech Republic. The students will get the opportunity to present at the Student Forum, which is a platform for graduate students at any career stage to introduce their research to the wider formal methods research community and solicit feedback. The field of formal methods is being rapidly deployed in a variety of areas both in academic research, as well as, in industrial systems. Thus, the broader significance and importance include fostering the next generation of researchers in this research area, as well as providing international experiences to build a globally aware workforce. In particular, students will have the opportunity to present at the Student Forum, learn state-of-the-art methodologies, be exposed to novel techniques, and interact with senior researchers in their areas of expertise. The organizers will give priority to students from under-represented groups and from universities without a formal methods program. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-07

PROJECT SUMMARY This grant application seeks funding for the 2024 Santa Cruz Developmental Biology Meeting, which will be held on the campus of University of California at Santa Cruz from August 23-27, 2024. This meeting is a biennial, grass-roots meeting organized by and for the scientific community working at the cutting edge of developmental biology. The Santa Cruz meeting, which has run continuously since 1992, oc- cupies a unique niche by combining international reputation with a relatively small size (~150 attendees) and a wholly new line-up of invited speakers at each gathering. As detailed in our application, we have planned for considerable participation by graduate students and postdocs by including short talks, posters, two work- shops aimed at career issues, and a career-perspective talk from a prominent scientist whose storied career pathway epitomizes the multi-disciplinary nature of developmental biology. The meeting format is based around single-platform sessions and three non-overlapping poster sessions so that all participants are engaged with the same topic and activity for the entirety of the meeting. The theme for 2024, “Unifying Principles of Organismal Development”, seeks to highlight common themes as well as top challenges facing the field, both conceptual and technical, as we seek fresh insights into the complex mechanisms underlying organismal development, tissue renewal, and the evolution of new forms. One chief goal of this meeting will be to bring together diverse researchers in an informal setting to highlight work across a broad spectrum of relevant systems, fostering cross-interaction between research fields and synthesis of new ideas. A second major goal will be to provide a forum for a diverse group of undergraduates, graduate students, and postdoctoral fellows to engage with and present their work to leaders in the field through talks, poster sessions, and informal discussions fostered by an isolated and relaxed campus setting. Along with a distinguished panel of four senior keynote speakers, the seven platform sessions will feature scientists working with a wide range of model organisms to address fundamental and emerging questions that form the basis for modern developmental research. Topics include: Cell-cell Communication, Theory and Modeling in Development, Active Matter and Mechanics, Convergent and Divergent Morphogenesis, Cellular Transitions and Plasticity, Information Processing & Gene Regulatory Networks, and New Technologies and Synthetic Approaches. The discussion that will ensue among the conferees will enhance our inclusive, multidisciplinary developmental biology community, stimulate new ideas, and accelerate basic developmental research and progress toward clinically relevant applications.

NIH Research Projects · FY 2025 · 2024-07

Project summary Postpartum hemorrhage (PPH) is the leading global cause of maternal morbidity and mortality. Uterine atony, insufficient uterine contractility after placental delivery, causes 80% of PPH. Oxytocin is the medical standard of care for prophylaxis and first line treatment of PPH. However, the mechanism whereby oxytocin causes uterine contractility remains incompletely understood. There is a critical need to determine (1) how oxytocin causes uterine contractility, and (2) the cellular and molecular mechanisms that underlie compromised oxytocin response in uterine atony. Such knowledge is integral to the discovery of novel, targeted treatments for uterine atony. Recent data in mice highlight a role for the Transient Receptor Potential Vanilloid 4 (TRPV4) channel in oxytocin- mediated uterine contractility. To address human uterine contractility, Dr. Ansari proposes innovative translational research using uterine tissue and derived smooth muscle cell cultures from cohorts of parturients with normal contractility and uterine atony. She will uncover molecular and cellular mechanisms of human uterine contractility in normal physiology versus uterine atony, define the role of the TRPV4 channel, and explore whether pharmacologic activation of TRPV4 may represent a novel uterotonic strategy. The first aim centers on normal, physiologic contractility, and the second on the pathophysiology of uterine atony: Aim 1) Determine the functional relationship between oxytocin and TRPV4 and the mechanisms of TRPV4 activation in normal uterine contractility; and Aim 2) Define TRPV4-dependent and TRPV4-independent mechanisms underlying uterine atony through cellular and molecular assays including single nuclei RNA sequencing. Dr. Ansari’s training will be conducted under the experienced mentorship of Dr. David N. Cornfield (smooth muscle biology, calcium signaling, and single cell transcriptomics), and Dr. Virginia Winn (maternal fetal medicine, basic and translational research in obstetrics), in conjunction with expert advisors in TRPV channel biology, biobanking, uterine atony, and PPH. This training will be enhanced by the Chan Zuckerberg Biohub Physician Scientist Fellowship program, advanced coursework in translational medicine, single cell RNA sequencing, and research laboratory management, and the supportive infrastructure of Stanford’s Maternal- Fetal Medicine and Obstetric Anesthesia Divisions. Through this K23, Dr. Ansari will obtain broad-based knowledge and skills in laboratory research including advanced experiential training in uterine smooth muscle biology, calcium signaling, and single cell transcriptomics. These skills will equip her to pursue her long-term goal to develop an independent research career identifying new therapeutic targets for uterine atony at the bench with the ultimate goal of translating discoveries to clinical care to address PPH.

NIH Research Projects · FY 2025 · 2024-07

Project Summary/Abstract Colorectal cancer (CRC) is the second most common cause of cancer-related death in the United States. Despite advances in both diagnostic and therapy, a large inter-individual variability in therapy response and diagnostic accuracy can be observed. The optimization of medical imaging agents in CRC is a critical component for improving our technical capabilities relating to early diagnostics, complete labeling of tumor-positive tissue for surgical resection, surveillance, and precision therapy. One major requirement for improving the clinical performance of therapeutics and medical imaging agents is the characterization of structural and functional chemistries in cancerous tissue that are either different from or absent in normal tissue. This is especially important in regions that may not be sufficiently labeled by existing imaging agents with a clinically acceptable and implementable signal-to-background ratio. In these cases, curative surgical intervention achieves mixed success in the CRC patient population due to the technical inability to label accurately and precisely tumor- positive tissues. Moreover, these differences may potentially explain the clinically observed variation in patient therapy response and disease aggressiveness. To develop a computational workflow that integrates CODEX imaging with high-dimensional hyperspectral chemical mapping and test my hypothesis, my experimental structure is divided into two main components. (1) I will perform preliminary non-destructive, label-free spatiochemical imaging on normal and CRC patient biopsies to identify differences in the functional heterogenous chemistries. (2) I will perform spatial omic identification of CRC related tissue architectures using CODEX for integration with the high-dimensional label-free 2D infrared spectral maps. The innovation of this research is the development of the computational workflow and framework that integrates highly multiplexed CODEX with high-dimensional label-free 2D hyperspectral chemical imaging. This approach can lead an alternative representation of physicochemical tissue properties for the characterization and identification of spatiochemistries relevant to CRC and its tumor-invasive front. Long-term, this can be used in the future to guide the development and optimization of cancer imaging agents to improve patient overall outcomes.

NIH Research Projects · FY 2026 · 2024-07

Project Summary Parasitic worms constitute a major global public health burden, with approximately one-fifth of the human population harboring one or more species within their gastrointestinal tract. We discovered that rare chemosensory epithelial cells called tuft cells sense parasites and initiate a type 2 immune response in the small intestine. Subsequent investigations into tuft cells at other barrier tissues throughout the body showed these cells respond to various microbial and environmental triggers. However, despite the massive density of microbes in the colon, the activation of tuft cells and their influence on type 2 immunity in the large intestine remains poorly understood. Several studies demonstrated key differences between tuft cells in the small intestine and colon, including distinct receptor profiles and divergent differentiation requirements. Recently, we found that commensal tritrichomonad protists in the microbiome are far more diverse than previously appreciated and stimulate colonic tuft cells and type 2 immunity independent of the tuft cell agonist, succinate. In Aim 1, we will examine the impact of tritrichomonad-tuft cell interactions on colonic immunity and determine the mechanisms colonic tuft cells use to initiate inflammation within the large intestine. Aim 2 will identify the novel tuft cell receptor(s) activated by tritrichomonads in the colon. Aim 3 will investigate the impact of colonic tuft cells on barrier function, epithelial protein secretion, and mucosal resistance to enteric pathogens. The results of this study will provide new insights into the microbial surveillance mechanisms of tuft cells in the colon and their influence on mucosal immunity. This knowledge will also reveal the potential for commensal protists to protect from enteric infections and provide a basis for using colonic tuft cells to treat inflammatory disorders in the large intestine.

NIH Research Projects · FY 2026 · 2024-07

Project Summary Typhoidal Salmonella serovars, including Salmonella enterica serovars Typhi (STy) and Paratyphi A (SPTyA), are the culprits behind devastating human-exclusive infections known as enteric fever, causing immense global morbidity and mortality. These pathogenic microbes adeptly outmaneuver the immune system by thriving in macrophages (MΦs), a critical aspect of Salmonella’s virulence. However, our understanding of how these typhoidal Salmonella strains interact with MΦs remains limited, with most research focused on non-typhoidal S. Typhimurium (STm). This gap in knowledge spurred our investigation. To bridge this gap in knowledge, we’ve employed unbiased methodologies, including random barcode transposon-site sequencing (RB-TnSeq) library screens under infection relevant conditions, genomic comparisons across different Salmonella serovars, and transcriptional profiling of MΦs infected by STy. These approaches have allowed us to identify typhoidal-specific fitness traits and factors. Our preliminary findings suggest that essential genes governing metal homeostasis contribute to typhoidal Salmonella’s fitness in MΦs due to pseudogenization of genes functioning in similar pathways. We have designed experiments to test these hypotheses (Aim 1). Furthermore, our bioinformatics analyses have led us to a putative typhoidal-specific virulence factor, which is translocated into MΦs via a type 3 secretion system (T3SS) and is vital for intracellular replication. We will take molecular and biochemical approaches to characterize this unique virulence factor within human MΦs (Aim 2). Finally, our comparative transcriptional profiling of MΦs containing replicating vs. non-replicating STy has unveiled a distinctive phenotype for MΦs containing replicating bacteria – anti-inflammatory, M2-like MΦs marked by phosphorylated STAT3 (pSTAT3). Elevated pSTAT3 levels in STy-infected MΦs are T3SS-dependent, suggesting that STy effectors manipulate the metabolic and immune landscape of human MΦs. We are determined to uncover specific virulence factors employed by STy to shape MΦ phenotypes (Aim 3). In essence, our research delves into the intricate strategies employed by typhoidal Salmonella to manipulate the host and inform new strategies for combating this formidable pathogen effectively.

NSF Awards · FY 2024 · 2024-07

Chlorine disinfection of drinking water has played a critical role in preventing outbreaks of waterborne diseases. Unfortunately, chlorine also reacts with dissolved organic matter in water supplies to form chlorinated byproducts. Consumer exposure to these byproducts is associated with slightly increased health risks. Drinking water regulations thus balance the need to reduce consumer exposure while maintaining effective disinfection. In the past, these regulations have focused on controlling the accumulation of small molecule end products that form as chlorine progressively breaks down dissolved organic matter. However, recent research has indicated that the larger byproducts formed during the process may be more important drivers of toxicity. The goal of this collaborative study by a chemist and a toxicologist is to compare the concentrations and contributions to toxicity of the large intermediate byproducts compared to smaller end products. The study will target water samples collected from drinking water utilities from different sources that use different disinfection techniques. The results of this project will be used to develop guidance to utilities on which disinfection conditions and techniques result in the formation of byproducts contributing to toxicity. The results also will benefit society by helping regulators identify byproducts that can serve as improved metrics of consumer exposure, thus contributing broadly to public health. Drinking water regulations have focused on trihalomethanes (THMs) and haloacetic acids (HAAs) as metrics of disinfection byproduct (DBP) exposure since the 1970s. Research over the past two decades has shifted to other unregulated 1-2 carbon DBPs such as haloacetonitriles that may contribute more to the toxicity of disinfected waters due to their greater cytotoxic and genotoxic potencies. While 1-2 carbon DBPs accumulate as terminal products, the higher molecular weight intermediate DBPs formed from the initial chlorine reactions with organic matter have received less attention. Recent research has indicated that these intermediate DBPs contribute more to the toxicity of disinfected drinking water. This project will compare the concentrations and contributions to toxicity of several novel classes of intermediate DBPs (halogenated phenols, proteins, and lipids) versus 1-2 carbon DBPs under different disinfection scenarios. The first objective is to compare DBP classes during chlorine/chloramine disinfection versus granular activated carbon (GAC) treatment followed by chlorine disinfection. The second objective is to assess how the formation of these DBP classes is affected by water age and water source (e.g., pristine source water, algal-impacted water, and wastewater-impacted water). The third objective is to assess whether nitrifying biofilms in chloraminated distribution systems increase the formation of these DBP classes by emitting DBP precursors. A fundamental assumption behind the current use of THMs and HAAs as metrics of DBP exposure is that their formation correlates with the toxicity drivers in disinfected waters. This study will test this assumption. The results will benefit society by providing information to regulators to assess whether alternative metrics of DBP exposure are needed to more accurately assess toxicity and whether efforts to reduce THM/HAA concentrations effectively reduce health impacts associated with DBP exposure. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-07

This project aims to develop an integrated photonic platform called MOLINO (Multilayer Oxide-clad LIthium Niobate On-chip) for efficient and compact generation of mid-infrared frequency combs. Frequency combs are precise tools for measuring different colors of light and have revolutionized metrology, spectroscopy, and sensing. However, current mid-infrared frequency comb sources are limited by high cost, low efficiency, and large size. By leveraging recent advances in lithium niobate nanophotonics and ultrafast laser technology, this project will create chip-scale mid-infrared frequency comb sources that require orders-of-magnitude less power than current systems while having much broader bandwidth and lower cost. If successful, this project will dramatically expand accessibility to mid-infrared frequency combs, accelerating scientific discovery and enabling new technologies to monitor greenhouse gasses, analyze chemicals, and sense biomolecules for health monitoring. This project will also train graduate students in cutting-edge skills in ultrafast optics and nanophotonics, supporting a diverse high-tech workforce. The collaboration between US and Swiss research teams will strengthen international scientific ties and leverage complementary expertise to maximize the project's impact. Technical description: The goal of this project is to develop the MOLINO platform, a chip-scale platform for generating broadband mid-infrared frequency combs pumped by ultrafast near-infrared lasers. This will be achieved through a collaboration between teams at Stanford University and University of Neuchâtel with three synergistic thrusts: (1) developing advanced GHz-repetition-rate, ultrashort-pulse-duration lasers in the 1-2 µm band based on Yb and Tm gain media; (2) creating multilayer-clad, dispersion-engineered lithium niobate nanophotonic waveguides and resonators with low loss, high nonlinearity, and low sensitivity to fabrication imperfections; and (3) demonstrating broadband, coherent mid-infrared light generation in MOLINO devices through nonlinear optical effects such as supercontinuum generation, difference frequency generation, and synchronously-pumped optical parametric oscillation. The teams will leverage their world-leading capabilities in ultrafast solid-state laser development, ion-beam-sputtered optical coatings, thin-film lithium niobate nanofabrication, and characterization of nonlinear photonic devices. The Swiss team will focus on laser development and coating fabrication, while the US team will lead MOLINO device design and fabrication, and the teams will work together to demonstrate and characterize the nonlinear dynamics of the MOLINO devices. If successful, this project could extend the bandwidth, efficiency, and repeatability of chip-based mid-infrared frequency comb sources by orders of magnitude beyond the state-of-the-art. This collaborative U.S.-Swiss project is supported by the U.S. National Science Foundation (NSF) and the Swiss National Science Foundation (SNSF), where NSF funds the U.S. investigator and SNSF funds the partners in Switzerland. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

The mission of the Stanford CTSA is to transform clinical, translational, and science of translation research and education (CTSRE) at Stanford and across the CTSA consortium. The Stanford CTSA’s programs in CTSRE have been designed to extend from the earliest stages of the translational pipeline to the “final mile” of implementation science taking full advantage of the rich research and educational resources available at Stanford University. Stanford CTSA’s organizational structure, by design, promotes interdisciplinary educational and research opportunities and stimulates collaborative innovation and discovery. The vision of the Stanford CTSA is to a) accelerate the discovery and translation of treatments; b) increase substantially clinical trial and clinical research participation; and c) achieve this vision by leveraging real-world data systems and AI approaches to optimize translational research and the science of translation. In the next seven years, we believe the Stanford CTSA can best achieve this by capitalizing on our greatest strengths, namely a) training the next generation of clinical, translational, and science of translation researchers; b) implementing programs in the principles of the science of translation that will span all elements of our CTSA; c) developing, optimizing, and implementing innovative data science and health informatics infrastructure, methods, and tools to promote analysis of large-scale real world data to serve as the platform for the conduct of CTSRE, accelerating the translation of discoveries into improved health; d) sustaining, developing, enhancing, and implementing infrastructure, methods, services, and tools to support our entire CTSRE efforts locally and across the CTSA Hub; e) imbuing all Elements of our Stanford CTSA with science of translation principles, including rigor and reproducibility (SPORR) and team science, in support of our CTSRE mission. Finally, we aim to develop and employ innovative state-of-the- field evaluative approaches to determine the effectiveness of all our CTSRE efforts. We aim to do this locally, regionally, and nationally in collaboration with our networks and across the CTSA consortium.

NIH Research Projects · FY 2026 · 2024-07

Motivation: Wireless technology has largely bypassed MRI. In an era of wireless earbuds and cell phones, MRI receive arrays remain hindered by bulky cabling and the commensurate RF baluns, and coax coupling that degrade performance. The simple act of adjusting the cable routing inside the MRI bore is a commonplace nuisance and source of lost time for patient handling. The ultimate goal of this proposal is to cut the cord - to demonstrate fully functional wireless receive array technology. This capability will open new avenues for array design and applications. It will become possible to create an open interface standard that is vendor- agnostic. Through wireless operation, arrays can finally be made wearable, without fear of mechanical fragility of the connections, thereby bringing the full SNR potential of wearable arrays to fruition. Approach: To architect MRI wireless arrays, we will develop all the necessary support technology, including wireless power transfer, low-power balun-free MRI receiver electronics, wireless coil Q-spoiling, ultra-wide band microwave short-range serial links, and microwave phase and motion synchronization systems. Notably, we can achieve these goals without need of custom IC fabrication for wireless interfaces. We will also leverage the compressibility of MRI k-space data by factors of 3x or more, to stream MRI data over multiple band-limited microwave channels. Moreover, we will leverage these added microwave fields to support motion, respiratory and cardiac sensing, to effectively provide non-contact vital signs sensing as a free byproduct of the wireless approach. The wireless arrays will be constructed and compared to geometrically equivalent topologies, for image quality and safety. The net outcome will be a wireless array with data transmission recovered and streamed over gigabit ethernet links to standard network interfaces. This approach will provide excellent synergy with recent efforts to make wearable lightweight conformable MRI receive arrays. Significance: The result of this project will overcome a common irritant and inhibitor of MRI receive arrays by removing the cable interface. This will enable wearable arrays to achieve their full imaging potential, while providing an interface that is vendor neutral, safer, and cheaper.

NIH Research Projects · FY 2025 · 2024-07

1 PROJECT SUMMARY 2 Coronary artery calcium (CAC), typically measured on gated computed tomography (CT) scans, is the 3 strongest predictor of atherosclerotic cardiovascular disease (ASCVD) events across all populations, including 4 groups underrepresented using traditional risk assessment methods. Despite the predictive ability of CAC 5 imaging, less than 1 million gated CT scans are performed annually in the US vs 19 million non-gated chest CTs 6 performed for reasons other than to measure CAC. Further, underrepresented individuals undergo fewer gated 7 CAC scans because these scans are not typically covered by insurance. When scored manually, non-gated CAC 8 scores predict ASCVD events as accurately as gated CAC scores, but grading CAC severity by visual estimation 9 is qualitative and inconsistent, and reporting varies substantially. Consequently, there is vital, lifesaving 10 information that has been collected but not used to guide preventive interventions for individuals unaware of their 11 increased ASCVD risk. 12 Stanford has developed a deep learning (DL) algorithm that quantifies CAC Agatston scores accurately on 13 routine non-gated chest CTs. We have shown that notifying patients and their clinicians about the presence of 14 incidental CAC dramatically increases statin prescriptions. The Novel Incidental Calcium Evaluation (NICE) 15 study will apply the now FDA-cleared algorithm on non-gated chest CTs performed across 3 geographically 16 diverse health systems (Stanford, MedStar Health, and Mayo Clinic). The study will include ~186,000 diverse 17 patients with non-gated chest CT scans without known ASCVD and follow-up within the 3 health systems. 18 Our team has expertise in preventive cardiology, radiology, epidemiology, health equity, DL, and qualitative 19 methods. First, the NICE study will evaluate the prevalence, epidemiology, and prognostic value of DL-CAC in 20 predicting ASCVD events across a real-world, diverse primary prevention cohort who underwent non-gated chest 21 CTs (Aim 1). Second, the algorithm will be extended to estimate CAC on lung cancer screening low radiation 22 dose CT scans (Aim 2). Automating CAC quantification would allow for the equitable and efficient implementation 23 of joint lung cancer and ASCVD screening programs for the 14.5 million eligible individuals in the US. Third, in 24 partnership with the National Minority Health Alliance, we will conduct focus groups with 100 diverse patient and 25 clinician stakeholders to identify facilitators and barriers to increasing preventive therapies following notification 26 of DL-CAC (Aim 3). 27 NICE will provide compelling evidence to support the immediate implementation of opportunistic screening for 28 and notification of CAC by leveraging routine, non-gated chest CTs already performed for other reasons. This 29 study fulfills the promise of data science approaches to equitably improve cardiovascular disease prevention.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT The candidate’s long-term career goal is to lead an independent research program focused on defining mechanisms that govern gut microbiome-host interactions in chronic heart failure and to translate this knowledge into novel preventative and therapeutic targets. This research program will pave the path for gut microbiome-focused precision medicine approaches in heart failure. The candidate will use the structured environment of the K08 award to acquire advanced research expertise in microbiome meta-transcriptomic and time-series analyses, integrative multi-omic data analysis, clinical research methods, as well as targeted professional development skills in mentorship, scientific communication, grantsmanship, and leadership. This will be accomplished in the context of investigating longitudinal gut microbiome-host interactions in chronic heart failure and how these relate to disease pathophysiology and progression. The candidate will leverage a longitudinal multi-omic and clinical dataset from a cohort of 60 patients with chronic heart failure with reduced ejection fraction, previously recruited by the candidate as part of her NHLBI-funded F32. The first aim will examine the relationship between gut microbial Bifidobacterium- based functional guild and heart failure severity and outcomes. The second aim will elucidate shifts in the host immune profile, gut microbiome composition and function, and heart failure disease status following a targeted Bifidobacterium probiotic intervention. A new cohort of patients with stable chronic heart failure with reduced ejection fraction will be recruited for this aim. This project will uncover mechanisms by which the gut microbiome interacts with different facets of host biology, which is expected to provide novel insight into disease pathophysiology and explain heterogeneity in heart failure progression. This knowledge will translate into better disease risk stratification and identify new therapeutic targets in heart failure. The proposed project will be performed under the guidance of an expert mentorship team comprised of world- leading experts in host-microbiome multi-omics (Dr. Snyder), mechanistic microbiome research (Drs. Sonnenburg and Tang), and heart failure translational research and clinical trials (Drs. Tang and Khush). They will provide complementary expertise in skills applied in this project and support the candidate's career development. The rich multidisciplinary environment at Stanford is optimal for this research proposal and provides an ideal setting for the candidate’s development into an independent researcher. At the culmination of this proposal, the candidate will gain the necessary expertise and preliminary data for an R01 proposal to identify synergistic microbiome-host pathways that underlie heart failure pathophysiology. As an independent physician-scientist, the candidate will use gut microbiome-based precision medicine approaches to improve the outcomes of patients she cares for as a heart failure physician.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Alcohol misuse negatively affects relationships and is significantly correlated with higher rates of relationship conflict, distress, and dissolution as well as other serious negative interpersonal consequences (e.g., domestic violence, sexual assaults). Encouragement from concerned partners (CPs) is a common motivator for those who misuse alcohol to pursue care and often the most helpful mechanism in supporting change. The goals of this proposal are to: Identify how specific CP behaviors influence their partner’s alcohol craving, motives, drinking, and problems on a daily basis using dyadic ecological momentary assessment techniques (Aim 1); use the knowledge from EMA analysis to iteratively develop a CP-focused web-based intervention (WBI) that provides psychoeducation about communication patterns that influence DP drinking and by integrating personalized feedback about CPs’ own communication behaviors that may be working against their goals (Aim 2); and pilot the WBI’s efficacy on CP outcomes (depression, anxiety, social support), their partner’s drinking behavior (alcohol consumption, motives, related consequences), and both partners’ relationship distress and conflict (Aim 3). We expect the WBI will yield significant improvements in all outcomes. This project is significant because intervening with CPs has strong potential to change relationship dynamics that may reduce problems and prevent future problems associated with alcohol misuse. It also develops a new prevention model that does not rely on the drinking partner attending a clinical facility to access care. The proposed study is innovative because it uses dyadic and ecological momentary assessment designs to test dynamic questions about interdependence in relationship interactions and alcohol use between partners and employs the generated knowledge to inform intervention adaptation. Teaching CPs to effectively communicate their concerns may be a necessary catalyst for decreasing their partner’s alcohol use and preventing alcohol use disorders. The potential reach of this intervention is large such that it can be easily implemented over the web to those who may need help but would not otherwise seek care.

NIH Research Projects · FY 2024 · 2024-07

PROJECT SUMMARY Varicose veins (VV), which affects approximately 23% of adults in the USA are non-lethal however, it negatively impacts the quality of life in many patients due to its debilitating symptoms. With limited methods for prevention and early treatments for VV, there is an unmet need to understand the mechanisms of VVs. VV pathogenesis is multifactorial and is a combination of epigenetic and genetic factors. In addition, hemodynamics and inflammation play an important role in venous pathology. Vein wall dilation occurs concurrently or as a consequence of the venous hypertension and inflammation, further exacerbating the venous reflux. These changes results in the overexpression of matrix metalloproteinases (MMPs), which causes degradation of the extracellular matrix proteins, thereby further affecting the structural integrity of the vein wall. This effect is due to changes in the endothelium (ECs) and smooth muscle cells (SMCs) is the alterations in cross talk which plays an important role in venous constriction. Alterations in venous tone also contribute to the development of VV. Our group identified 30 independent genetic variants associated with VV in a previous genome-wide association study (GWAS). In complex disease traits, the disease-associated loci may be in non-coding regions even if they may be responsible for gene expression regulation. It is difficult to know in which cell types or physiological contexts this regulation occurs. Thus, candidate genes require validation to establish causality. One way of doing this is with single-cell genomic assays. In our study, we propose to elucidate the molecular mechanisms behind the development of varicose veins by leveraging the recent advances in single-cell sequencing technologies. By using single cell RNA-seq, we propose to examine the transcriptomic changes at the single cell level in varicose veins. Moreover, we propose to decipher the key regulators effecting the different components of the varicose vein and their effects on ECSMC crosstalk (Aim 1). Due to limited access to ECs and SMCs from patients (given that the isolation and long-term culture of vascular cells from patient’s blood is an extremely difficult task), the mechanisms underlying vascular dysfunction in varicose veins remain largely unknown. As part of Aim 2, we will utilize the induced pluripotent stem cell technology to recapitulate the disease phenotype of varicose veins in vitro.

NSF Awards · FY 2024 · 2024-07

Cities across the US have experienced a significant increase in people experiencing homelessness, especially since the beginning of the COVID pandemic. Timely and early intervention that improves the well-being of those who are experiencing homelessness significantly improves their outcomes, reduces time spent in homelessness, and prevents persistent homelessness. However, because of the dynamic movements of unhoused persons (due to clearing of encampments, weather, safety, etc.) coupled with a reluctance to provide information to the authorities, it is difficult for existing programs to determine the magnitude and location of service needs and to ensure that well-intentioned programs do not inadvertently reduce overall wellbeing. The project will support research that will measure neighborhood conditions and factors that impact the wellbeing of homeless populations through cameras, noise, and environmental sensors mounted on cars driving throughout the city of San Jose. This data will help determine neighborhood conditions at a granular level and the localized need of the homeless population and to optimize the services they receive (e.g., meal delivery, trash and waste removal, and toilets) through our partners including the City of San Jose, Loaves & Fishes, and Feed My Lamb. The project has four main technical research steps to achieve the goal of understanding neighborhood wellbeing and the local needs of the homeless population: (1) developing a community-driven vehicular and mobile crowdsensing system to measure neighborhood conditions, (2) designing clustered federated learning algorithms to reconstruct city-wide maps of neighborhood environments and service needs, (3) modeling the causal relationships between neighborhood environments and wellbeing across different communities, and (4) developing methods to optimize services to improve and reduce inequality in wellbeing. The research project involves three types of community partners: local food pantries, local residents, and the city government of San Jose. Through collaboration with these partners, the project will have immediate impact to provide localized actionable needs relating to food, trash, and toilets, and to improve the wellbeing of vulnerable populations in San Jose, CA. The methods and models developed in the project will be generally applicable to other cities and areas with diverse neighborhoods. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2024-07

Project Summary A molecular pathway that links N-glycosylation to birth defects Developmental defects are the leading cause of both fetal loss before birth and infant deaths after birth. The immense advances in our genetic understanding of mammalian development over the past four decades have not yet translated into better preventative or therapeutic options for babies with birth defects. Our ability to address this high unmet need depends critically on the discovery and detailed mechanistic understanding of molecular and cellular pathways that drive human developmental defects. Working with a multidisciplinary investigative team that combines expertise in signal transduction, human genetics and bone biology, we have discovered an novel inter-organelle communication mechanism that links protein N-glycosylation in the Endoplasmic Reticulum (ER) to the reception of WNT signals at the cell surface. The WNT/β-catenin pathway is a key cell-cell communication system that regulates tissue patterning during development and regenerative responses in adults. Human genetic studies from our collaborators show that this ER-based pathway is disrupted in a severe, deforming subtype of the inherited bone fragility disorder Osteogenesis Imperfecta (OI) and in subtypes of Congenital Disorders of Glycosylation (CDG), characterized by developmental defects across tissues. Given that over 25% of proteins encoded in our genomes are N-glycosylated, we propose the existence of a surveillance mechanism (analogous to the unfolded protein response) that ensures WNT-driven differentiation is only allowed to proceed if the ER N-glycosylation machinery is intact. The three goals of this proposal are to delineate the components and transduction mechanisms of this regulated N-glycosylation pathway, understand its role in osteoblast differentiation and bone matrix production and establish its relevance to human birth defect syndromes. We use genome-wide CRISPR screens and mass spectrometry to identify pathway components, gene-editing to disrupt or mutate these components in both cell lines and primary cells from human OI patients, and mechanistic studies to understand how N-glycosylation in the ER tunes WNT ligand sensitivity at the cell surface. Successful completion of this project will define a new pathway that regulates WNT signaling and human development by using N-glycosylation as a regulatory post-translational modification. More broadly, our work will define a largely unexplored signaling function for N-glycosylation, a fundamental cell biological process linked to diseases across multiple organ systems.

NIH Research Projects · FY 2025 · 2024-07

Progress in treating neurologic disease depends on the recruitment, retention, and support of neurosurgery and neurology physician scientists (NSci). Our major goal is to increase the number of NSci residents who transition to independent academic careers, tackling complex neurologic questions with translational impact. We will achieve this through early multidimensional support of trainees, creating a stimulating and collaboration-fostering environment. We will focus on early engagement in research training, individualized and enduring mentorship of the UE5 scholars, and iterative program design, to foster the strongest pool of NSci physician scientists. To this end, we will establish an innovative UE5 training program which extends beyond the architecture of the residency training years, to increase the recruitment of candidates, and ensure their retention and successful transition to independent research careers. These innovations include: (1) integration with established Stanford pathway programs to engage future physician scientists, (2) Day 1 enrollment of NSci residents into an UE5-foundational research training pipeline; (3) tiered, cross-departmental leadership, functioning throughout the continuum of NSci training to ensure the success of the UE5 trainees; (4) increasing trainee independence through establishment of an UE5-to-K Transition Team, and (5) continuous improvement with outside feedback from an External Advisory Committee. This adaptive UE5 educational structure will identify, equip, and encourage NSci Residents immersed in direct investigative study, increasing retention and reducing time to a mentored or independent research award. Our active, engaged commitment to NSci residents will ensure they are well-prepared for independent research careers, and directly benefit the trainees, patients, and scientific communities we serve.

NSF Awards · FY 2024 · 2024-07

Neural networks have revolutionized science and engineering in recent years, but their theoretical properties are still poorly understood. The proposed projects aim to gain a deeper understanding of these theoretical properties, especially the statistical ones. It is a matter of intense debate whether neural networks can "think" like humans do, by recognizing logical patterns. The project aims to take a small step towards showing that under ideal conditions, perhaps they can. If successful, this will have impact in a vast range of applications of neural networks. This award includes support and mentoring for graduate students. In one direction, it is proposed to study features of deep neural networks that distinguish them from classical statistical parametric models. Preliminary results suggest that the lack of identifiability is the differentiating factor. Secondly, it is proposed to investigate the extent to which neural networks may be seen as algorithm approximators, going beyond the classical literature on universal function approximation for neural networks. This perspective may shed light on recent empirical phenomena in neural networks, including the surprising emergent behavior of transformers and large language models. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-07

Aging is the primary risk factor for debilitating diseases such as Alzheimer’s disease. Can manipulation of neurons in the brain alter the body’s physiological state to extend lifespan and prevent neurodegenerative disease? Neuropeptides are signaling molecules released by neurons that act through modulatory receptors expressed throughout the brain and body to regulate homeostasis. Whether neuropeptides could control long-term phenotypes such as the rate of aging, neurodegeneration and cognitive decline remains largely unknown. Neuropeptides have been implicated in Alzheimer’s disease in humans. For example, the neuropeptide Galanin (GAL) is overexpressed in degenerating brain regions in Alzheimer’s disease, low levels of the neuropeptide Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) are correlated with higher amyloid burden and memory decline, and the number of neurons expressing the neuropeptide Hypocretin are significantly reduced in postmortem hypothalamus of Alzheimer's disease patients. However, a systematic characterization of the role and mode of action of neuropeptides in regulating vertebrate lifespan and their impact on neurodegeneration and cognitive decline is missing. This is largely because aging and lifespan experiments in transgenic vertebrates are slow (3+ years in mice) and low throughput. I will take advantage of a short-lived vertebrate model the African killifish to (1) determine if deletion of key neuropeptides can alter lifespan, healthspan, and cognitive decline, (2) investigate the mode of action of one neuropeptide that I have already found to extend lifespan when knocked out, and (3) test if neuropeptides can act as pro-longevity factors when delivered later in life to counter age-associated cognitive decline. To achieve this, I will use interdisciplinary technologies at the nexus of genetics, aging, and neuroscience. I already have exciting tools and data that support my goal. I built a library of neuropeptide knockout killifish targeting 22 human-conserved neuropeptides using CRISPR/Cas9 and I optimized the protocol for lifespan and healthspan assessment in the killifish. In tantalizing preliminary data, I found that knockout of the AD-associated neuropeptide GAL in killifish results in progressive cognitive decline suggesting that neuropeptides could be key modulators of neurodegeneration in disease such as Alzheimer’s disease. By focusing on diverse neuropeptides that interact with specific druggable receptors, I hope the long-term impact of this work will translate to clinical solutions to age-associated Alzheimer’s disease and others. For the mentored part of my training at Stanford University, I will receive training from my mentor Dr. Karl Deisseroth, co-mentor Dr. Anne Brunet, and an exceptional scientific advisory team with expertise in neuroscience, neuropeptides, aging, neurodegeneration, genetic screening, and CRISPR methods. This work, my technical training, and my career development at Stanford University will provide me with the skills and foundations required to be a leader of a laboratory at a top academic institution, discovering genes critical for longevity and for countering cognitive decline in Alzheimer’s disease.