Stanford University

NIH Research Projects · FY 2025 · 2022-08

C-DIAS OVERALL: PROJECT SUMMARY/ABSTRACT This is a revised proposal for a NIDA P50 Center of Excellence: The Center for Dissemination and Implementation At Stanford (C-DIAS). Our overarching goal is simple to say, yet paradigm shifting if achieved: Equitable access to evidence-based addiction treatments. C-DIAS unites established addiction dissemination and implementation (D&I) research experts with complementary areas of strength. C-DIAS Research Core members bring the best of current approaches to D&I science in measures, methods, design and modeling, as well as in health economics and policy translation. Three coordinated Research Projects serve as study vehicles to improve D&I science in addiction by reducing variation and increasing standardization in measures and methods, translating and harmonizing data across studies, and using agent- based and economic modeling to scientifically respond rather than merely react to substance-related epidemics and health care disparities. In addition to serving as the C-DIAS organizational hub, the Administrative Core will provide a platform to build expert capacity in D&I research in addiction, offering a stratified range of Training Education and Consultation Services, with open access to online resources and tools for the broadest possible audiences. C-DIAS will advance D&I science by greater precision and rigor in evaluating implementation strategies— which are the “interventions” of an endeavor to install or scale up evidence-based treatments. Proven pharmacological or psychosocial addiction treatments are the “what” of any implementation effort, whereas implementation strategies pertain to “how” these interventions are most effectively translated and brought to scale. Policymakers and systems leaders seek empirically-based tools to make informed decisions about not only what to implement, but how to do so effectively and at what cost. C-DIAS is based at Stanford but designed to serve as an open, vibrant and highly visible national center without walls. It will leverage established relationships and generate new connections among independent multi-disciplinary teams of individuals across geographically dispersed and diverse entities. C-DIAS will develop human capital, foster synergy, and accelerate professional, scientific and public health impact with outcomes otherwise unachievable in its absence. We propose a sorely needed and radical approach, with one critically significant and urgent goal: To improve public health by making the best addiction treatments equitably available for those most in need.

NIH Research Projects · FY 2024 · 2022-08

ABSTRACT. Glucose homeostasis plays a critical role in multiple cellular processes, and impaired or altered glucose metabolism is associated with a wide range of pathological states. A key step in glucose metabolism is catalyzed by the glycolytic enzyme pyruvate kinase. Proliferating cells almost universally express the pyruvate kinase M2 (PKM2) isoform, which can assume either an active or inactive state. PKM2 is at the nexus of cellular metabolism, and determines whether cells metabolize glucose into ATP or use it to make more of the necessary building blocks for cell division. Multiple studies have demonstrated how dynamic changes in PKM2 expression contribute to altered glucose metabolism in different contexts. The ability to non-invasively visualize and track dynamic changes in PKM2 expression will enable improved understanding of altered glucose metabolism and the downstream mediators of glycolysis in multiple disease states. The lack of PKM2 expression within the brain and myocardium make this imaging strategy highly promising for neurological and cardiovascular applications. We have recently reported the development and human translation of [18F]DASA-23, the first clinically-relevant and specific radiopharmaceutical to detect, localize, and quantify PKM2 using positron emission tomography (PET) imaging. We have determined the biodistribution, radiation dosimetry, and brain distribution of [18F]DASA- 23 in healthy volunteers, and have explored its ability to visualize PKM2 expression in one potential application of patients with primary brain tumors. Although our results highlight the potential of imaging PKM2, [18F]DASA- 23 has several limitations that impedes widespread use, and the ability to study PKM2-mediated glycolytic reprogramming in broader applications. This includes high radiation dose to the gallbladder wall, a high degree of non-specific binding within white matter in the brain, and poor solubility in radiotracer formulation vehicle. This proposal will develop novel PKM2 radiotracers to overcome the limitations of [18F]DASA-23. Development of a safe and reliable PKM2 radiotracer will enable repeat assessment of the dynamic alterations in glucose metabolism in multiple different applications and patient populations. We will establish the synthesis and fluorine- 18 radiolabeling of two candidate small molecules with improved physicochemical properties relative to DASA- 23, pharmacological activity and specificity for PKM2, and the potential for radiolabeling. We will automate the radiosyntheses and characterize uptake and specificity in cell culture (Aim 1), determine biodistribution and radiation dosimetry (Aim 2), and assess the ability to visualize PKM2 expression in one potential application of primary brain tumors (Aim 3). Success of this proposal will develop novel radiotracers for visualizing a hallmark of metabolism. This will have important ramifications for studying altered glucose metabolism in multiple applications and could improve our collective understanding of metabolic adaptations in disease. Importantly, this technology will be adopted by a wide range of users in different pre-clinical and clinical studies.

NIH Research Projects · FY 2026 · 2022-08

PROJECT SUMMARY / ABSTRACT The goal of this proposal is to identify novel coinhibitory checkpoint molecules operating orthogonally to currently targeted checkpoints in cancer patients, such as PD-1. Despite the unprecedented success of checkpoint blockade immunotherapy as a therapeutic approach against multiple types of cancer, significant inter-individual variability remains, ranging from no response to complete remission of systemically metastasized lesions. Various tumor-intrinsic parameters, such as tumor mutational burden, neoantigen burden, and mismatch repair pathway deficiencies, only show limited predictive power for favorable clinical outcomes. Therefore, complementary to such tumor-intrinsic factors, it is plausible to hypothesize that humans have hitherto unknown coinhibitory mechanisms orthogonal to currently known immune checkpoints and that a minor fraction of humans with germline or somatic mutations in such pathways could benefit from exceptionally potent antitumor immunity triggered by checkpoint blockade immunotherapy. Identification of such orthogonal pathways would significantly enhance the health of patients with multiple types of cancer; for instance, the 15 types of cancer for which pembrolizumab has been indicated. The current project aims at dissecting the human genetic and immunological basis of coinhibitory crosstalk between human T cells and cancer cells by tackling the two specific aims. Specific Aim 1 is to delineate the molecular mechanisms of PD-L1/L2-mediated inhibition of T cell activation independent from PD-1. This aim is supported by preliminary observations that both primary and immortalized T cells from a patient with inherited complete PD-1 deficiency, as well as PD-1-negative human T cell lines Jurkat and HuT78, all responded to co-inhibition by bead- immobilized PD-L1/L2. CD83 was identified as a candidate for PD-L1-mediated immunosuppression. The proposed work includes searching for additional candidates through RNA sequencing and surface receptor profiling and functionally validating the candidates via RNA silencing and CRISPR-guided knockout. Specific Aim 2 is to characterize the immunoregulatory function of CD83 in human T cells. Previous studies suggest that CD83 can serve as both an immunosuppressive receptor and ligand for human T cells, although its counterreceptor(s), downstream signaling, and relevance to antitumor immunity remain mostly elusive. The proposed work includes biochemical characterization of CD83 signaling in human T cells with or without cancer cell coculture utilizing genetic knockout and lentiviral rescue of CD83 gene. The proposed project will help untangle the biochemical and immunological basis of human T cell coinhibitory circuits in the crosstalk between cancer cells and also substantially bolster the applicant's scientific portfolio and expedite the growth toward an independent investigator pioneering human genetics of cancer immunology and immunotherapy.

NIH Research Projects · FY 2026 · 2022-08

Genomic instability is a characteristic hallmark of cancer, leading to an increased propensity for mutations in tumor suppressor genes, activation of oncogenes, and chromosomal alterations. These genomic changes not only contribute to tumor initiation but also play a critical role in tumor progression and the response to anti-cancer treatments. Targeted therapy aims to trigger cancer cell death by disrupting cell cycle pathways, while also exploiting the high levels of genomic instability present in cancer cells. In small cell lung cancer (SCLC), the use of the WEE1 kinase inhibitor AZD1775 exhibits potent anti-cancer activity by disrupting the G2/M checkpoint, allowing damaged cells to undergo mitosis, eventually leading to mitotic catastrophe. However, despite its promising anti-tumor effects in clinical settings, AZD1775 resistance emerges rapidly, indicating the development of genomic instability, acquired gene alterations, or compensatory mechanisms to bypass cell death. A genetically engineered mouse model of breast cancer with WEE1 conditional deletion shows that WEE1 deficiency increases the activity of APC/C, allowing cancer cells to progress through mitosis, impairing genomic integrity and resulting in carcinogenesis. Moreover, gene expression analysis in SCLC patients revealed PKN1 as a highly expressed cell cycle-related gene that is involved in G2/M delay in a WEE1-low expression group. These observations lead us to hypothesize that WEE1 inhibitory-mediated genomic instability is a driving factor in tumor resistance, promoting the acquisition of neo-oncogenic events and compensatory mechanisms in the G2/M checkpoint, enabling SCLC cells to evade cell death when treated with WEE1 inhibitors. Given the significance of genomic instability in cancer resilience, this K00 project aims to investigate whether WEE1 inhibition promotes acquired genomic instability and gene mutations that drive resistance while exploring the potential role of PKN1 in WEE1 inhibitor-resistant SCLC.

NIH Research Projects · FY 2026 · 2022-08

The biggest biomedical challenge of this century is the restoration of diseased organs and tissues. Unlike humans, salamanders have the extraordinary ability to rapidly regenerate organs, including limbs, spinal cords, hearts and brains. Our goal is to discover how these animals rebuild functional adult tissues in a matter of weeks. From development through degeneration – the health and function of our organs depends on production of appropriate tissue-specific proteins. Yet, our current understanding of regeneration is largely based on studies of mRNA and not on direct assessment of proteins that are ultimately required for repair. This is in part due to technical limitations – microarray and RNA-Seq technologies revolutionized our understanding of transcription- but until recently we lacked the tools to study translation of mRNA into protein at the same scale and resolution. The Mexican axolotl is famous for its lifelong “youthfulness”. Axolotls share with other salamanders the surprising and incompletely understood ability to regrow entire limbs after amputation. By combining cutting-edge methods in translation research, we were able to demonstrate that, unlike in mammals, severe injury in the axolotl surprisingly results in rapid activation of protein synthesis at a time when there is little cellular proliferation. This unusual molecular response is a feature specific to regenerative vertebrates and relies on activation of the mammalian target of rapamycin (mTOR) pathway. Moreover, we find that remarkably fewer than 20% of all axolotl mRNAs are translated at any given time, the remainder exist in a ‘free’ state outside the translation machinery. We will test the hypothesis that the ‘free’ transcripts in the axolotl may be spatially organized into membrane-less compartments comprised of functionally-related and translationally co-regulated mRNAs and that transcripts critical for cell survival and cell fate specification shuttle between these compartments and the ribosome to facilitate wound healing and regeneration. We have further identified that control of protein synthesis at the time of regeneration is highly dependent on the ability of the Axolotl to surpass a stress activating signal and instead promote activation of the mTOR pathway. We will test the hypothesis that the structural/sequence specific differences in Axolotl mTOR components can shed light on functional differences in upstream regulation of protein synthesis between species and the remarkably ability to repurpose a ‘stress-response’ signal to a ‘growth and regeneration’ signal. These findings suggest the possibility that poor healing in mammals may be due to a distinct cellular signaling response at the site of injury rather than to an inherent lack of regenerative potential. Lastly, we have found that amputation of the limb in the axolotl triggers selective translation of some ribosomal proteins but not others, coincident with the “burst” in protein synthesis. We will therefore test the bold hypothesis that axolotls may assemble distinct subsets of specialized ribosomes to facilitate selective expression of transcripts critical for wound healing and regeneration. Together, this proposal seeks to provide a novel mechanistic understanding as to why some species can regenerate while others cannot.

NIH Research Projects · FY 2025 · 2022-08

Project Summary – Overall Bone marrow produces blood cells whose functions range from oxygen delivery to anti-microbial defense to hemostasis, all originating from hematopoietic stem cells (HSC). To sustain and regulate this process, bone marrow stromal cells form multiple niche microenvironments, each tailored to the needs of a particular developing blood cell population. Using highly-multiplexed imaging technologies, our proposed Bone Marrow Tissue Mapping Center (TMC) aims to systematically and quantitatively dissect the cellular composition and spatial organization of human bone marrow microenvironments. The resulting detailed maps will serve as an open and global platform for understanding which cells and interactions are critical for each branch of hematopoietic maturation, and how these vary by anatomical site and across diverse patient demographics. The TMC will define cellular identities and cell states at the transcriptional, translational, and post- translational levels using Nanostring DSP, Multiplexed Ion Beam Imaging (MIBI), and MALDI-MSI, which generate quantitative spatial maps of RNA, protein, and N-glycans, respectively. Our cross-disciplinary team not only includes the inventors of MIBI and a pioneer of MALDI-MSI, but also experts in human HSCs and human hematopoiesis and a practicing hematopathologist with expertise in histopathologic bone marrow diagnosis. To overcome the unique challenges of working with hard, mineralized bone, we will leverage parallel, robust, clinically-validated bone marrow processing pipelines which maximize and standardize sample quality and compatibility with current and future technologies. Integrating seamlessly into standard clinical workflows, our pipelines enable convenient sharing of prospectively-collected materials with the Tissue Core. Samples will be collected from three different sources: (1) prospective, patient-matched multi-site collection from deceased donors to examine differences between anatomical sites, (2) prospective collection of femoral head from hip arthroplasty specimens for differences between age ranges, (3) iliac crest bone in the Stanford Pathology archive for differences between races and genders. These multiple collection strategies, multiple sites, and different investigational focuses complement prior HuBMAP projects. The Data Analysis Core team has pioneered multiple novel data processing pipelines, including pixel-based analyses, cell-based analyses including state-of- the-art cell segmentation and cell clustering and enumeration, and neighborhood analyses. These tools are broadly-applicable to all highly-multiplexed quantitative imaging technologies. Overall, our team and strategy are exceedingly well-suited for executing the vision of the proposed Bone Marrow TMC. The spatial structure of bone marrow reflects the evolutionary mechanisms that terraformed bone to create unique microenvironments meeting the nutritional needs of developing blood cells with divergent functions. The interdependence between bone marrow tissue structure and hematopoiesis is informative not just in blood cell maturation, but for understanding metabolism, aging, and development of cellular therapies.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT Non-coding elements comprise 98% of the human genome. The coordination of non-coding regulatory elements in the mammalian genome plays a pivotal role in controlling gene expression. Both experimental and computational studies reveal that pathogenic genes involved in complex diseases, including oncogenes, are regulated by a large number of enhancers, implying the existence of a complex interdependent regulatory network of enhancers in modulating and maintaining expression of these genes. Genome-Wide Association Studies (GWAS) reveal that non-coding regulatory elements, including enhancers, are hotspots for the genetic predisposition to disease. To determine causal relationships between chromatin architecture and gene transcription, perturbation in a biological system is necessary. Recent advances in CRISPR-based genome engineering and live cell imaging technologies have enabled new techniques for ultrahigh resolution interrogation of the function of various genome regulatory elements and how they relate to gene expression. In preliminary studies in our lab, we performed a targeted CRISPR interference (CRISPRi) based screen to study how the 7 MYC enhancers present in K562 cells work together to co-regulate this oncogene. We created a library with >87,000 pairs of gRNAs targeting the MYC enhancers to understand the epistatic network of gene regulation underlying MYC expression. We found that when a subset of enhancer pairs were targeted together, they exhibited a more dramatic than expected reduction in growth rate. We developed a model that divides MYC enhancers into 2 layers that work together with varying degrees of efficiency to co-regulate MYC expression in K562 cells. Here, we seek to expand these preliminary results to examine additional oncogenes and perform these experiments in additional cell types. In addition, we will combine perturbation of oncogene enhancers with CRISPR-based live cell imaging (termed CRISPR LiveFISH), that allows for the dynamic imaging of multiple genomic loci, mRNA, and protein components in living cells. In Aim 1, we will develop an ultrahigh-resolution multiplexed CRISPRi/a tiling screens platform to dissect enhancer interactions of different oncogenes in different cancer cell lines. We will perform multiplexed CRISPRi/CRISPRa screens to inhibit or activate pairs of enhancers with an ultrahigh spatial resolution (~20bp) controlling four oncogenes (MYC, CCND, BCL2, PDE4DIP) in K562 and HeLa cells. In Aim 2, we will characterize the dynamic real-time interactions between transcriptional coactivators, mediators, multiple enhancers, promoters, and RNA transcription during CRISPRi/a-mediated perturbation. We will monitor real-time dynamics of different enhancers, promotors, RNA transcription, and the transcriptional coactivator proteins BRD4, IRF1, and Gata4 using LiveFISH with and without enhancer perturbation. Altogether, we seek to apply new CRISPR technologies developed in our lab to create a model of how oncogene enhancers are dynamically regulated across multiple oncogenes and in multiple types of cancer cells.

NIH Research Projects · FY 2025 · 2022-08

The combination of lineage tracing and single cell RNA sequencing (scRNAseq) in mouse atherosclerosis models has created a paradigm shift in our understanding of vascular disease, showing that lesion smooth muscle cells (SMC) undergo phenotypic transitions into derivative cells with multiple complex phenotypes. We identified TCF21 as a coronary artery disease (CAD) associated gene mapped by genome-wide association studies (GWAS) and showed that this gene regulates a disease-related transition of SMC to a fibroblast like phenotype, producing cells we term “fibromyocytes.” Further, we and others have shown that medial SMC can also transition to a second SMC-derived cellular phenotype, characterized by expression of genes known for their role in endochondral bone formation, substantiating and expanding previous work investigating this process that is linked to intimal vascular calcification. We showed that this chondrogenic process, which gives rise to cells we term “chondromyocytes” (CMC), is actively inhibited by two CAD associated genes, one encoding the TGFB1 signaling molecule SMAD3, and the other encoding the environmental sensing aryl hydrocarbon receptor (AHR). Knockout (KO) of both genes in mouse models showed increased transition to CMC, larger lesion size and increased vascular calcification. These studies identified SOX9 as a primary driver of the phenotypic transition to the CMC phenotype. Our longterm goal is to elucidate the molecular mechanisms that mediate the detrimental CMC transition. Our Central Hypothesis postulates that SOX9 is a key initiator of this chondrogenic process in the vascular wall, as it is in endochondral bone formation, and regulation of its expression and function in SMC is intimately linked to vascular calcification and disease risk. Our objective is thus to determine the upstream epigenetic signals that modulate SOX9 expression, and how SOX9 expression contributes to CMC development and vascular calcification. Specifically, in Aim 1 we will employ Sox9 KO and SMC lineage tracing in the ApoE KO mouse atherosclerosis model to characterize the effect of this gene on SMC cell state transitions, and the impact of perturbing these transitions on disease morphology and cellular anatomy. In Aim 2, we will conduct scRNAseq in these mice to characterize the SMC gene expression program downstream of Sox9 in this cell type. Single cell assay of transposase accessible chromatin sequencing (scATACseq) in the same animals will map enhancers genome-wide that are differentially regulated in CMC phenotypic transition, and identify specific transcription factors (TFs) that bind these enhancers to regulate expression of CMC genes. Studies proposed in Aim 3 will employ in vitro studies to characterize the transcriptional and epigenetic mechanism by which SOX9 interacts with the inhibitory factors SMAD3 and AHR, and novel TFs that promote transition to the CMC phenotype. The proposed studies will identify cellular and molecular mechanisms that mediate SMC transition to CMC, and the relationship of this process to vascular calcification and disease risk.

NIH Research Projects · FY 2026 · 2022-08

PROJECT SUMMARY Reconstruction of the urinary tract in urologic surgery oftentimes involves utilizing the small intestine to replace a segment of the urinary system, and as a result, patients may suffer from the side effects of connecting the urinary and gastrointestinal tracts. Patients that undergo urinary diversion surgery, for example, are at risk of infection, electrolyte abnormalities, and ileus as a result of this practice. If a better substitute for the urinary tract were available, outcomes from urinary diversion or reconstructive surgery involving the small intestine would be drastically improved. The main goal of this proposal is to develop a source of autologous urothelial stem cells that can potentially be used towards the development of alternative bladder or urothelial substitutes. In this proposal we hypothesize that urothelial stem cells can be generated via direct conversion of fibroblasts and can reconstitute the bladder urothelium in the mouse. First, we will aim to generate urothelial stem cells via direct conversion, or transdifferentiation (Aim 1), and we will achieve this by overexpressing transcription factors associated with bladder development and screening for suprabasal and basal urothelial markers. We will validate our screening results with functional assays with organoids as well as multilayered assembloids. Second, we will map the epigenetic changes that take place during urothelial stem cell differentiation to suprabasal cells (Aim 2), and we will accomplish this by performing Omni ATAC-seq on control bladder organoids and urothelial stem cell organoids in basal and differentiation media conditions. By identifying differences in areas of chromatin hyperaccessibility, we will be able to identify transcription factor binding motifs enriched in stem cell and differentiated cell states. Finally, we will develop a bladder urothelial stem cell transplant protocol using mouse models of urothelial ablation and injury (Aim 3). We will determine if urothelial stem cell transplantation can reconstitute all cell types within the urothelium utilizing urothelial stem cells obtained from mouse bladders as well as those obtained from transdifferentiation, and we will test for functional outcomes. If we are successful in these aims, we will demonstrate that autologous urothelial stem cells can be generated via direct conversion of fibroblasts, and we will establish a source of cells for bladder substitute tissue engineering as well as a basis for cell-based therapy for disorders of the urothelium that are typically treated with surgical reconstruction using the gastrointestinal tract, such as severe radiation cystitis or severe interstitial cystitis. The impact of this work on human health will be significant as this work will potentially make urinary diversion and reconstruction surgery a much less morbid surgical option for patients with severe urothelial disorders.

NIH Research Projects · FY 2024 · 2022-07

Late preterm (34-36 weeks gestational age) infants account for 7% of the 3.76 million live births in the United States annually, or over 263,000 infants each year. Compared to term infants, late preterm infants are at increased risk of morbidity from outcomes such as hypoglycemia, temperature instability and hyperbilirubinemia, and often require medical intervention in a neonatal intensive care unit (NICU). Thus, while the vast majority of infants born at term stay with their mothers in a well infant (level I) nursery during the birth hospitalization, many late preterm infants are instead hospitalized in the NICU where they may be separated from their mothers. However, significant variation exists amongst hospitals for NICU admission rates and clinical thresholds for admission in late preterm infants that is not explained by clinical illness. Preliminary data obtained by the PI suggests that institutional criteria for requiring automatic NICU admission in late preterm infants can vary from 34-37 weeks gestational age and 1500-2500 grams birth weight. This represents late preterm infants of varying maturity and size, and likely does not precisely capture infants who are at highest risk of needing NICU level interventions. The goal of this proposal is to identify optimal NICU admission criteria for late preterm infants. A large retrospective cohort of late preterm infants born at a single institution will be assembled, collecting data on admission locations, and occurrence and management of late preterm morbidities. With this, Aim 1 will be addressed: identify the frequency of neonatal morbidities amongst infants born at 34-36 weeks’ gestation, and the frequency of these morbidities requiring medical intervention. Literature on the frequency of morbidities in late preterm infants is limited, and none currently exists delineating the proportion of these morbidities that require clinical intervention. Subsequently, in Aim 2: a prediction model will be developed for which late preterm infants are most likely to benefit from automatic admission to a NICU at the time of birth. The cohort generated in Aim 1 will be utilized to compare clinical parameters of infants who required at least one NICU level intervention to those that did not require any. Training and test data sets will be established. Using cross-validation techniques within the training set, an optimal cut-point for a score derived from the predictive model will be chosen to drive clinical decision-making based on the sensitivity and specificity of the decision rule. The strategy will be evaluated on a test set. The obtained prediction model will be a resource towards informing optimal NICU admission criteria for late preterm infants. The PI will train in study design methodology, data analysis, modeling, and grant writing during this fellowship that will advance her career path towards an independent physician scientist focused on identifying high value care practices that safely promote an intact mother-infant dyad in newborn care. She will benefit from the world-class research and clinical environment, and renowned expertise at Stanford University.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY Dr. Bollyky is a mid-career investigator who has developed a successful, translational research program focused on bacteriophages – viruses that infect bacteria - and bacterial wound and airway infections. He is also a highly productive research mentor, serving as a director of the physician-scientist training program for the medical residency program as well as training numerous residents, fellows, and medical students in his own lab. The goal of this K24 is to provide Dr. Bollyky with protected time to grow his patient-oriented research program and mentor additional clinical fellows, junior faculty and other trainees in patient-oriented research. The proposal also provides for dedicated time and resources to help Dr. Bollyky enhance his own mentoring skills, obtain scientific and career guidance from an advisory committee, sharpen his skills in vaccinology and microbiology, and expand his knowledge and technical skills in human clinical studies. The scientific focus of this work is bacteriophages – viruses that infect bacteria – and their impact on bacterial infections. Phages are abundant in the human body but their impact on human health is largely unknown. The Bollyky lab has recently reported that phages produced by the major bacterial pathogen Pseudomonas aeruginosa (Pa) promote chronic Pa infections. In particular, they have identified a novel mechanism by which filamentous Pf phages produced by Pa contribute to antibiotic tolerance by functioning as structural elements in Pa biofilms – slimy communities of bacteria and polymers that allow bacteria to colonize airways and other surfaces. Pf phages organize host and microbial polymers in ways that produce a robust biofilm that resists penetration by antibiotics, leading to antibiotic resistance. Their team recently published that Pf phages are associated with heightened antibiotic resistance in patients with cystic fibrosis (CF), a genetic disease associated with thick, tenacious sputum and chronic lung infections with Pa. It may be possible to protect against Pa infection by targeting Pf phages. The Bollyky Lab recently developed a vaccine that targets Pf phages to prevent Pa infections. However, it is unclear whether antibodies against Pf phages naturally occur and whether these are protective against Pa infection. Here, Dr. Bollyky will test the hypothesis that Pf phages contribute to antibiotic resistance and chronic infections while Pf antibodies protect against this. In Aim 1 they will define how Pf phages contribute to antibiotic tolerance in Pa biofilms. In Aim 2 they will define how Pf antibodies protect against Pa infections. Finally, in Aim 3 they will determine whether Pf and anti-Pf phage antibodies influence clinical outcomes in patients with CF. Together, these studies will give rise to novel therapeutic targets and treatment strategies against Pa biofilm infections and launch the careers of multiple young physician scientists. Protected time for career development and mentoring will allow Dr. Bollyky to broaden the scope and influence of his and his trainees’ work and help to sustain and grow the patient-oriented research enterprise of the NIAID and the NHLBI.

NIH Research Projects · FY 2026 · 2022-07

Mitochondrial malfunction is well known to manifest as dysfunction of neurons, due to the high energetic demands of these highly polarized cells, their remarkable axonal length, and complexity. However, how neuronal mitochondria precisely control crista structure in response to ever- changing energy demands and oxidative stressors and how these regulations impact neuronal function and behavior in multicellular organisms remain elusive. In our Preliminary Studies, we have discovered an evolutionarily conserved role for MIC60 in maintaining crista structure in Drosophila. We have found molecular modifications of fly MIC60 (dMIC60) including phosphorylation and oxidation and their significance in maintaining crista junction plasticity and resisting oxidative stress. These results demonstrate the vital importance of dMIC60 and its posttranslational modifications for mitochondrial and neuronal homeostasis. We hypothesize that molecular regulations of dMIC60 including phosphorylation and oxidation allow dMIC60 to receive cellular signals to conduct its functions and that their misregulations could lead to neuronal dysfunction and degeneration. To test this hypothesis, we will perform the following Specific Aims. Aim 1: Roles for dMIC60 in neuronal homeostasis. Aim 2: Crista structure as a cellular cause for oxidative damage. Aim 3: Molecular mechanisms of dMIC60 phosphorylation and oxidation.

NIH Research Projects · FY 2026 · 2022-07

Project Summary This application for a Mentored Patient-Oriented Career Development Award (K23) will allow the candidate to obtain specialized training and research expertise in new areas that will enable her to become an independent physician scientist. Dr. Fischer details a five year research plan aimed at characterizing the effects of cannabis exposure on neurobiological functioning and the evolution of depressive symptoms in transition age youth (TAY). The specific aims are to prospectively examine the impact of cannabis exposure on reward and stress system functioning, and to develop a preliminary integrative biomarker-guided model of the effects of cannabis exposure on symptoms of reward impairment and depression in TAY. Forty regular cannabis-users and 30 non-cannabis- using controls will be recruited and prospectively followed for 12 months. Participants will complete clinical and substance use assessments, perform reward and stress tests utilizing functional neuroimaging and cortisol bioassays, and provide hair samples to quantify aggregate delta-9-tetrahydrocannabinol (THC) exposure. Findings from this work will advance our understanding of the neurobiological and psychiatric consequences of cannabis exposure in TAY. The application builds on the candidate’s prior research experience in characterizing the acute effects of cannabis in dual-disorder patients and examining functional connectivity biomarkers of risk, resilience and treatment response in depression. Over the course of the K23, Dr. Fischer will receive in-depth mentored-training in new areas that are essential to her career development: (1) the multidimensional characterization of reward processing and impairment; (2) the neurobiology of phyto/endo-cannabinoids and the hypothalamic-pituitary-adrenal (HPA) stress axis; and (3) the longitudinal investigation and analysis of translational cannabis and depression research in TAY. An interdisciplinary team of mentors who are experts in the characterization of reward function and impairment in relation to substance abuse (Dr. Knutson), phyto/endo- cannabinoid pharmacology and physiology (Dr. Piomelli), HPA stress-axis physiology and pathophysiology in depression (Dr. Schatzberg), adolescent cannabis research (Dr. Tapert), and statistical methods for analyzing longitudinal data (Dr. Jo) will help the candidate meet her training and research objectives. This program of research and training directly aligns with NIDA and NIMH Strategic Plans and Priorities by rigorously characterizing the neurobiological and psychiatric impact of cannabis exposure during an understudied period of brain development using quantitative, multivariate, and longitudinal assessments. Dr. Fischer’s research and training will take place at Stanford University. This K23 Career Development Award will provide a fundamental foundation for the candidate to achieve her career goals of characterizing the longitudinal effects of cannabis exposure on neurobiological function in relation to clinical symptoms, thereby informing public poloicy and assisting in the development of neurobiologically-guided interventions for cannabis use and depressive disorders.

NIH Research Projects · FY 2026 · 2022-07

PROJECT SUMMARY (See instructions): What operations are performed by the mammalian central nervous systems to coordinate and conduct voluntary movement? Motor systems neuroscience seeks to understand these neural mechanisms. The last two decades have witnessed a transformation in this field with the use of multielectrode recordings and statistical estimation and modeling techniques. These technological advances have yielded rich, low-dimensional neural dynamics that are suggestive of the mechanisms underlying behavior. To minimize confounds, the overwhelming majority of these studies utilize behavioral constraint to isolate just the behaviors of interest for study. While effective for generating many behaviorally similar trials, this may have the unintentional consequence of artificially constraining neural dynamics to a subset of its full range. This project seeks to better understand the full repertoire of behavior in a freely-moving setting and model this neural activity using novel computational tools. This project combines the expertise of two neuroscientists with complemenary skillsets spanning systems and computational neuroscience. This project will involve acquiring novel, freely-moving data using the recent advances in depth imaging cameras and modeling the dynamics of the labelled neural data with innovative switching dynamical models. TThis combined expertise will be applied to the investigation of the initiation vs sustaining of movement, the decomposition of walking periods into distinct dynamical regimes, and an analysis of foraging behavior. Taken together, these studies will further our understanding of how neural dynamics drive unconstrained motor behavior. This insight has implications for the development of ambulatory brain-machine interfaces and may inform the treatment of individuals with motor disorders such as stroke.

NIH Research Projects · FY 2025 · 2022-07

Tuberculosis (TB) is a respiratory disease that causes the death of 1.5 million people each year. Approximately 30% of the human population worldwide are latently infected with TB, creating a risk for developing active TB and transmission. Genomic studies have attempted to identify innate immune genes and polymorphisms that affect susceptibility to TB among humans. Despite findings that suggest correlation of certain genetic polymorphisms in specific immune genes with TB susceptibility, genomic studies have failed to identify specific isoforms that might influence human TB susceptibility. This is because innate immune genes are highly conserved among humans, making them exhibit similar immune responses to TB bacteria. By contrast, different species of nonhuman primates exhibit different levels of susceptibility to TB. Rhesus macaques (macaca mulatta) are more susceptible to TB compared to cynomolgus macaques (macaca fascicularis), yet the underlying mechanism of this variation is unknown. It is crucial to compare innate immune response genes across primate species in response to TB infection to identify the differences in immune orthologs that help eliminate TB bacteria. Using comparative genomics approaches, this study explores and characterizes immune responses across species by ex-vivo infection of different primate blood cells with TB bacteria using long-read RNA-Seq. The use of long-read RNA-Seq provides a more comprehensive annotation of innate immune orthologs across primate species which wasn’t possible in past studies using short-read RNA-seq. This study will help identify species-specific immune orthologs important in immunity against TB, which might ultimately be used to evaluate drug targets for human TB prevention and treatment. This comparative genomic approach can also be applied to identify species-specific immune orthologs as drug targets for other infectious diseases.

NIH Research Projects · FY 2026 · 2022-07

SUMMARY Opioids are the most effective analgesics but are associated with severe side effects including respiratory depression, tolerance, and addiction. These factors helped cause the opioid abuse epidemic in the US, making drug overdose the leading cause of accidental death in the US. Thus, the identification of safer analgesics with diminished side effects and abuse potential is critical to address the ongoing crisis. Clinically used opioids predominantly exert both their analgesic and adverse effects through their action on the µ-opioid receptor (MOR). While several approaches were taken towards safer analgesics, these efforts are limited by a lack of understanding the complex biochemical networks engaged and activated by MOR in response to ligand binding. This proposal builds on recent evidence suggesting that (1) MOR signaling is dependent on the interplay between subcellular localization and membrane trafficking in a ligand-specific manner and (2) MOR shows ligand- dependent effects on its protein interaction network and the signaling pathways it activates. Thus, delineating the MOR-initiated signaling pathways for endogenous peptides and addictive opioids and how these are coordinated by receptor location and trafficking provides potential new strategies for therapeutic modalities and safer analgesics. The overarching goal of this proposal is to combine quantitative proteomics, functional genomics, and opioid receptor biology to systematically discover and characterize regulators of MOR signaling and trafficking in human induced pluripotent stem cell-derived neurons. We will combine proximity labeling mass spectrometry and quantitative phosphoproteomics to systematically delineate interaction networks that MOR engages and map the signaling pathways it activates. To study the functional role of proteomic targets in MOR signaling and trafficking, we will develop and apply reporter assays for receptor signaling and trafficking in CRISPRi gene regulation screens. Finally, we will test mechanistic hypotheses from proteomic and genetic screens on how novel regulators of trafficking and signaling fine tune the cellular response of MOR activation. Our proposed approach will yield mechanistic insights into MOR-initiated signaling pathways and how these are regulated by receptor trafficking. Identifying key regulators of MOR activation will fill a critical gap for designing safer, pathway selective analgesics and treatments for opioid addiction.

NIH Research Projects · FY 2026 · 2022-07

PROJECT SUMMARY / ABSTRACT To optimally support global child health and development, research needs to be designed to account for the scale of disease burden and child mortality that occurs worldwide. A robust global pediatric health research strategy depends upon new investigators with the technical expertise required to translate basic science to population health and wellness. Although the US leads the world in biomedical research, none of the 39 NIH funded global health institutional training grants are focused solely on improving health for children in low- and middle-income countries, and none are from NICHD. General pediatric global health fellowships limit career opportunities for clinician scientists, further restricting researchers to high income health systems. We will capitalize on the significant teaching and research resources that exist across Stanford University to develop the next generation of pediatric subspecialty global health researchers. We propose a dual fellowship training approach which will encompass both the trainee’s pediatric subspecialty as well as a focus on global health approaches to subspecialty research. Resources such as the co-located seven schools which make up teaching and research at Stanford University, as well as the surrounding Silicon Valley, offers a unique entrepreneurial and technologically innovative environment, will serve as the hub of an interdisciplinary environment to build a cross-cutting research program addressing the needs of children in low- and middle-income countries. The program PI has directed T32 programs for over a decade and is the School of Medicine Senior Associate Dean for Faculty Development and Diversity, so is well-positioned to provide substantial mentorship and leadership resources for the trainees. Stanford has a group of highly experienced and well-funded global child health researchers with substantial mentorship and clinical experience who work across the spectrum of low- and middle-income countries. Finally, the proposal includes a unique pediatric subspecialty global health curriculum that integrates with existing rigorous coursework and a Master’s in Epidemiology program to provide the foundation for the proposed training program. Two Stanford pediatric subspecialty fellows will be selected per year, beginning in their second year of training. Trainees will: (1) complete a Master of Science in Epidemiology and Clinical Research; (2) receive structured dual-mentorship by both subspecialty and global child health researchers; (3) design and complete a hypothesis-driven research project; (4) attend interactive research working groups; and (5) participate in the Department of Pediatrics highly successful grant writing programs. This comprehensive program will train a new generation of pediatric subspecialty global health scientists equipped to advance the research agenda outlined in the NICHD Scientific Vision.

NIH Research Projects · FY 2025 · 2022-07

The prime objective of our multi-center National K12 “Diabetes-Docs: Physician-Scientist Career Development Program” (DiabDocs) is to support the research career development of the next generation of physician-scientists in basic and clinical diabetes for academic careers with a specific focus on type 1 diabetes and other forms of diabetes. Our structured K12 program is designed to provide a mentored research experience together with tailored career development training. We propose these aims to achieve the DiabDocs program goals: 1) Create a national cohort of up-and-coming physician-scientist researchers by developing cohesion through shared programmatic training cohort experiences including an annual retreat. 2) Expand the geographical and pipeline reach of K12 programs so that Scholars train locally while maintaining the historic strengths of the past institutional diabetes K12 programs. 3) Develop an outstanding national mentor community to train diabetes physician-scientists and provide a more visible pathway to research careers for medical students/residents/fellows by active recruitment programs and participation in the DiabDocs retreat and educational programs. 4) Ensure consistency and effectiveness of mentoring across research centers through shared mentor training and resources. In addition to our MPI structure, one of the distinguishing features of our program is that previously funded K12 Scholar program PIs and their experience are part of the DiabDocs Executive Leadership Committee complemented by the addition of two Adult Endocrinology physician-scientists with expertise in basic science, clinical science, and health services research. Monthly career development programming and local mentoring of K12 Scholars is supplemented by Scholar presentations at the DiabDocs annual retreat in the Spring and at the American Diabetes Association plus external ‘arm’s length’ mentors and a visiting K12 Scholars Program to engage experts at other institutions. The first 4 recruitment cycles (2 cycles in year 1 of grant) resulted in 72 letters of interest with 47 invited applications representing 28 unique institutions of which 24 were from institutions that did not previously have a diabetes related K12. With the start of Year 4 of DiabDocs we will have funded 24 K12 Scholars of whom 19 are from ‘new’ institutions. To date, 2 K12 Scholars have ‘graduated’ to their own K08/23 awards with 2 other awards in process and 9 additional K08/23 proposals submitted. Our K12 scholars will be well-positioned to serve as catalysts for multidisciplinary investigations bridging bench-to-bedside science leading to patient-centered discoveries. We are confident that we will continue to attract high caliber applicants and mentor them toward impactful careers in academic medicine as skilled physician-scientist leaders.

NIH Research Projects · FY 2025 · 2022-07

Project Summary/Abstract The objective of “Brain Computer Interfaces and Disability: Developing an Inclusive Ethical Framework (BCI- DEF)” is to use structured vignettes, video-supported interviews, and a deliberative democracy approach to assess and analyze diverse, critical stakeholder perspectives about the benefits, risks, and ethical challenges of Brain Computer Interface (BCI) technology. BCIs measure and interpret brain signals and interface with a device to allow users to perform a task, such as communication or movement. BCIs hold great promise, especially for end users with disability due to neurologic deficits in communication or motor function, but this potential may be limited by unaddressed ethical challenges. Issues of concern include stigma, autonomy, privacy, safety, impacts to personhood, responsibility and justice. Through the innovative approach in this project, we will focus on input from a primary group of potential end users (patients with neurologic disability and their family/caregivers) and clinicians who care for patients with acute neurologic injury to obtain critical and heretofore absent information about important focus areas related to ethics of BCI. The inclusion of additional key stakeholders including BCI engineers and research teams will further inform the deliberative democracy approach. Issues we will further investigate through this project include weighing the potential benefits and risks of BCI; evaluating ethical challenges surrounding BCI and disability including those around autonomy, stigma, and justice; impact of BCI on perceptions of disability and clinical decision making in acute brain injury; and defining critical information needed by end users, families, and clinicians to make future informed decisions about BCI use. This work is critical to ensure seamless, equitable, and ethical adaptation of innovative and cutting edge BCI technologies being developed through BRAIN Initiative projects.

NIH Research Projects · FY 2025 · 2022-07

Enter the text here that is the new abstract information for your application. As diabetes technologies have become more innovative and effective in the management of pediatric type 1 diabetes (T1D), research and usage has not engaged all youth living with T1D. Studies have consistently demonstrated 50% lower rates of diabetes technology use in youth of lower SES. Although diabetes technology has the potential to improve in pediatric T1D outcomes. This proposal aims to build an evidence base for data-driven interventions designed to increase uptake and utilization of diabetes innovations by addressing barriers and supporting promoters of diabetes technology use. Ananta Addala, D.O., M.P.H, is a physician scientist committed to a career as an independent investigator addressing factors associated with in T1D management and outcomes. Dr. Addala’s longstanding research and clinical interests are to promote care for youth with T1D. As a physician with a background in pediatric endocrinology, epidemiology, and behavioral health, Dr. Addala is uniquely qualified to address factors associated with diabetes technology use youth with T1D. Dr. Addala has enlisted a multi-disciplinary mentorship team comprised of experts in the fields of pediatric T1D, health disparities, statistics, and mixed method study design to successfully execute this proposal and launch an independent research career in pediatric T1D. The overall objective of this proposal is to discover factors associated with diabetes technology use in youth with T1D and public insurance and develop a brief intervention, as a means to understand and improve pediatric T1D outcomes. This will be accomplished through two aims. In aim 1, focusing on the family, Dr. Addala will construct an evidence base of barriers and promoters to diabetes technology use in youth with public insurance in order to formulate and test a brief pilot intervention aimed at increasing uptake. In aim 2, this time focusing on the providers, Dr. Addala will construct the evidence base on barriers and promoters to recommending diabetes technology to youth with public insurance in order to formulate and test a brief pilot intervention to increase provider recommendation of diabetes technology. Taken together, findings from Aims 1 and 2 will result in the development of an intervention aimed at increasing diabetes technology uptake and access in youth with public insurance, thereby improving T1D outcomes. Dr. Addala will use the K23 mentored award to execute an in-depth training plan which includes formal coursework and structured mentorship by her mentors to advance her understanding of mixed methods research, intervention development, and expertise on statistical methods. This proposal is foundational to a future independent clinical trial to evaluate the efficacy of the interventions developed on promoters and barriers of diabetes technology use in youth with T1D.

NIH Research Projects · FY 2026 · 2022-07

ABSTRACT During the past few decades, there has been a rapid increase in the incidence of oropharyngeal cancer (OPC), which is attributable to the epidemic of oral human papillomavirus (HPV) infection. Patients with HPV-positive OPC respond well to concurrent chemoradiotherapy and have a more favorable prognosis than HPV-negative patients. However, standard treatment is associated with significant toxicity and likely represents over-treatment for many patients with HPV-positive disease. Several randomized clinical trials have tested novel deintensification strategies with the goal to reduce toxicity and improve patients’ quality of life while preserving the high cure rate. These trials enroll patients based on cancer stage and smoking history. However, current clinical prognostic factors are rather crude and do not accurately predict disease progression on an individual level. Reliable prognostic models are critically needed for personalized risk-adaptive therapy of OPC. To address this unmet need, we propose quantitative CT features to characterize intratumoral spatial heterogeneity and disease invasion/spread, which are known drivers of treatment resistance and disease progression. In addition to knowledge-based image features, we will develop complementary data-driven deep learning models to predict disease progression by using a retrospective multi- institutional dataset of 1771 patients. Further, we will integrate imaging with clinical data to improve prediction and establish their validity by rigorous prospective validation in 1780 patients enrolled in 3 randomized clinical trials. Finally, we will employ a radiogenomic approach to elucidate biological basis of the imaging signatures. If successful, the proposed models will allow more accurate prediction of prognosis and improve risk stratification of OPC. This has significant therapeutic implications by optimizing the selection of patients for treatment deintensification, which will increase the likelihood of success of future clinical trials and pave the way for precision medicine in OPC. Because the information is derived from standard CT scans, this would be readily integrated into current clinical workflow, widely applicable to underserved populations in low-resource settings, and therefore would help reduce health disparity in the US.

NIH Research Projects · FY 2025 · 2022-07

Summary Our Genetics and Developmental Biology (GENETICSDEVBIO_T32) training program combines the expertise of multiple Stanford departments to provide rigorous and innovative training in genetics and developmental biology across a range of living systems. We believe that interdisciplinary training in genetics and developmental biology is particularly urgent in 2021 and beyond. High-throughput sequencing and massive studies of genetic variation are transforming much of biomedical research. However, connecting DNA sequences to traits remains a key challenge in all fields. Our training program directly addresses the problem of mapping genotypes to phenotypes through combined training in genetics, genomics, and experimental methods for testing gene functions in a wide range of organisms. The major training goals of the program are to: Provide students with rigorous 5-year Ph.D. training at the forefront of modern genetics, genomics, and developmental research. Develop fundamental skills in scientific thinking. Develop skills required to manage and analyze scientific data in a rigorous and reproducible manner. Train students to carry out scientific research ethically and responsibly. Train students to effectively communicate findings in written and oral formats. Advance basic and applied knowledge by training scientists who will consider a range of questions, populations, and study systems in their research. We achieve these goals through an innovative combination of a joint computational and experimental training camp; joint courses, retreats, seminar series, journal clubs, and research talks; formal and ongoing instruction in ethics and rigorous design of experimental research; and monthly T32-specific “professional skills” discussion meetings throughout graduate school. Our outstanding faculty and trainees have an exceptional track record of combining techniques across disciplines and making major scientific advances that improve our mechanistic understanding of basic biology and human health and disease.

NIH Research Projects · FY 2025 · 2022-07

PROGRAM SUMMARY / ABSTRACT Objective: The principal mission of the Stanford K12 Clinician-Scientist Career Development Program is to fund a premier clinician-scientist/clinician investigator training program, designed to leverage excellence in basic, translational and clinical research to develop new clinician-scientists who will emerge as productive, independent physician scientists and strong leaders across the eye and vision research community. Rationale & Design: Through this institutional career development award, we will incorporate a rigorous program with outstanding vision research faculty serving in various mentor and supervisor roles, comprehensive eye and vision courses, and significant clinical ophthalmologic exposure to facilitate future applications of vision research. The Byers Eye Institute along with the broader vision research community at Stanford University has a proven record of success in mentoring, along with sufficient external support to support all scholars. Key Activities in the Training Plan: In this program, participating faculty will lend their expertise by serving across Mentoring Groups, an Executive Committee, and an External Advisory Board to help build, review and support each Scholar's career development plan. As recent K-awardees, additional junior faculty will also serve in the Near-Peer Mentoring Committee to lend additional advice and guidance and build a community of mutual support for Scholars. Scholars will be partnered with mentors appropriate to their scientific focus and will be trained in basic or clinically relevant research, as well as additional essential skills meant to propel their transition to independence, such as statistics, scientific writing, grantsmanship, ethics, leadership and the responsible conduct of research. Each program will be customized for each Scholar according to their specific interests, skills and background, needs, and career goals. Planned duration of appointments and projected number of scholars: With an emphasis on recruiting strong applications from women and under-represented minority applicants, a maximum of 3 postdoctoral Scholars will be selected each year after their completion of a post-residency clinical fellowship in ophthalmology. K12 Scholars will typically spend 2 years in the program. Intended scholar outcomes: Successful outcomes will be measured by productivity in papers and presentations, successful transition to independent K awards and later conversion to R-funded research, and impact on the field of eye and vision research as well as clinical ophthalmology. Stanford has robust mentoring systems, strong curriculum, and solid infrastructure in place to prepare Scholars for research independence. Together, the program's myriad resources will create a supportive environment for K12 Scholars throughout the duration of the program.

NIH Research Projects · FY 2025 · 2022-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. This application will support and renew the Medical Scientist Training Program (MSTP) at Stanford University School of Medicine. In over 50 years of continuous NIH funding, over 350 trainees have graduated from this program, many of whom have become leaders in their fields of academic medicine and biomedical research. Our program provides a superb environment and unique advantages for fulfilling our mission “to train a talented cohort of pioneering physician scientist (PS) leaders dedicated to a lifetime of biomedical discovery that improves human health through innovation.” Dual degree pre-doctoral training in the Stanford MSTP has durably produced successful outcomes in our trainees, based on several metrics used to compare MD/PhD programs nationally, including low attrition and time to degree conferral, publication record, and high retention of graduates in biomedical research careers. This proposal describes important elements of the Stanford MSTP. Program enhancements include a significantly increased level of institutional support, reflected in increased staffing and faculty effort to direct the MSTP; increased financial support for student training by the School of Medicine; integrated training in rigorous, reproducible, responsible and safe research conduct throughout the curriculum; improved mentoring and monitoring mechanisms for students throughout their training; enhanced career development training and physician scientist community building; new physician scientist training tracks for MD-only students, and enhanced integration of MD-only trainees in MSTP activities. Together, the increased institutional support, increased effort by School of Medicine faculty and leadership, dedicated program enhancements, and integration of clinical and graduate training have changed, expanded and improved the Stanford MSTP. Support through this proposal, heavily leveraged with School of Medicine and University resources, will support continuing innovation in training physician scientists at Stanford.

NIH Research Projects · FY 2024 · 2022-07

PROJECT SUMMARY/ABSTRACT Research: RNA binding proteins (RBPs) regulate diverse cellular processes including transcription, translation, and regulation of gene expression, and are frequently dysregulated in cancers. Through an unbiased genetic screen aimed at identifying cancer-specific RBP dependencies, we recently identified a specific requirement for RBM39 in malignant myeloid cancers and that cancers bearing RNA splicing factor mutations as being particularly sensitive to the anti-cancer sulfonamides. RBM39 is an RBP that functions in RNA splicing and recently, a class of clinical-grade “anti-cancer sulfonamide” compounds were demonstrated to degrade RBM39 protein by co-opting the Ddb1/CUL4 ubiquitin-ligase complex as their mechanism of action. Thus, the primary goal of this project is to assess differential and tissue-specific requirements for RBM39 in normal hematopoiesis versus myeloid malignancies, and to assess requirements for RBM39 for leukemia initiation and maintenance. This proposal will utilize a novel conditional knockout (cKO) mouse for Rbm39 and several associated newly developed in vitro and in vivo murine models to pursue this goal. We expect these investigations to further our understanding of the role of RBM39 in normal physiology and cancer as well as provide new therapeutic insights into the on- and off-target toxicities of the anti-cancer sulfonamides. These goals are particularly timely given that several of these molecules have already proven excellent safety in multiple phase I/II trials and are now ripe for therapeutic testing in a patient population most likely to benefit from RBM39 degradation. Candidate: Dr. Sydney X. Lu is a graduating hematology & medical oncology fellow in the Department of Medicine at MSKCC. He aims to become an independent, tenure- track physician-scientist investigating the molecular pathogenesis of hematological malignancies through a combination of genetics, functional genomics, and murine modeling. Dr.Lu has outlined a five-year period of mentored training to strengthen his skills in functional genomics and disease modeling. This training period will be carried out under the mentorship of Dr. Omar Abdel-Wahab, a leader in the functional genomics of hematopoietic malignancies. Dr. Lu has also assembled an advisory committee composed of Drs. Ross Levine, Martin Tallman, Michael Kharas, and Christine Mayr who will help guide his training and research. Environment: MSKCC is the world's oldest and largest private cancer center, devoting more than 130 years to exceptional patient care, innovative research, and outstanding educational programs. MSKCC exposes trainees to an exceptionally robust academic research environment with a strong commitment and track record of successfully supporting junior faculty who are seeking careers as independent physician-scientists.