Yale University

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT This proposal aims to characterize the impact of GLP-1 analogue obesity treatment on mechanisms of modifiable cardiometabolic risk factors in young people with type 1 diabetes (T1D) complicated by obesity through assessment of adipose, glucose, and lipid physiology. Obesity and overweight impact 40% of adolescents and young adults with T1D, a population in whom T1D alone already drastically increases future cardiovascular disease risk. Our preliminary data indicate that regardless of BMI category, most adolescents with T1D have a visceral to subcutaneous adipose tissue ratio as high as youth with obesity and prediabetes. This visceral fat ratio in youth with T1D is proportional to the degree of hepatic insulin resistance. The study will comprehensively assess drivers of cardiometabolic risk factors in young people with T1D and obesity while examining the impact of GLP-1 analogue obesity treatment on visceral adipose tissue (the ratio of visceral to visceral+subcutaneous adipose tissue), insulin resistance, and postprandial lipemia. To achieve these aims, we will utilize 1) an abdominal MRI to quantify abdominal adipose distribution, 2) the stepped euglycemic hyperinsulinemic clamp with stable isotope tracers to assess insulin resistance and gluconeogenesis, 3) a DEXA scan to measure body composition, and 4) a high-fat mixed meal tolerance test to quantify postprandial changes in atherogenic lipoproteins. After completing these assessments, 54 young adults with T1D and obesity who meet the criteria for anti-obesity pharmacotherapy will be randomized to 1- year of treatment with oral semaglutide or placebo. The metabolic assessments will be repeated at 1-year of the treatment. A comparator group of 15 young adults with T1D and lean body mass index will undergo the metabolic studies at one-time point. Carrying out the proposed research program is critical to advance the understanding of the strategies to reduce cardiometabolic risk and impact the pathophysiologic mechanisms promoting cardiometabolic risk in young patients with T1D and obesity. This is consistent with the NIDDK's mission to improve the health and quality of life of individuals with diabetes. Results will reveal the intricacies of cardiometabolic disease risk and potential treatment in young people with T1D complicated by obesity.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY Neural activity in the neocortex is plastic over a range of temporal scales. Learning associations between sensory stimuli and behaviorally relevant outcomes drives cortical plasticity and is fundamental to an organism’s survival. Chronic stress can also impact neural circuits although its contributions to sensory cortex connectivity and sensory encoding is unclear. Changes in information processing in the neocortex can take place at different spatial scales: in local microcircuits made up of heterogeneous excitatory and inhibitory cell types and in larger interconnected cortical networks. Rodent studies from the last decade have revealed an elaborate network of secondary visual areas that may be involved in visually-guided behaviors such as associating initially neutral visual stimuli with aversive events. However, the dynamic network connectivity of these secondary areas and their distinct contributions to learned, visually guided fear behavior is unknown. Fear-learning enhances the cortical representation of stimuli that predict a foot shock, but it is currently unknown if different inhibitory elements support these changes in visual stimulus representations. Using innovative imaging approaches this proposal will address 3 Aims: (1) Determine the changes in network dynamics and functional connectivity that accompany fear learning. The proposed experiments will investigate primary and secondary visual cortical contributions to learned fear behavior using simultaneous dual 2-photon/widefield imaging and cortical inactivation approaches. (2) Determine the microcircuits that contribute to fear-learning related changes in cortical responses. Using cell-type specific 2-photon imaging and optogenetics Dr. Moberly will test the hypothesis that VIP-mediated disinhibition enhances cortical output neuron responses. (3) Investigate the consequences of stress for functional sensory cortical network architecture and its relationship to ongoing behavioral state. Dr. Moberly will conduct this research in the labs of his mentors Drs. Jessica Cardin and Michael Higley at the Yale University School of Medicine with input from advisory committee members, Drs. Marina Picciotto and Michael Crair. In the K99 period, Dr. Moberly will learn new technical skills in cellular 2-photon and simultaneous dual 2-photon/widefield imaging in combination with optogenetics and quantitative behavioral approaches. The proposed experiments and multifaceted training plan will impart Dr. Moberly with a unique combination of skills that will position him to transition into a successful independent career as a systems neuroscientist.

NIH Research Projects · FY 2025 · 2023-09

Project Summary/Abstract Precious metal catalysts are typically used for the synthesis of active pharmaceutical ingredients (APIs) even though first-row transition metals such as Ni are more sustainable and can facilitate unique reactivity. For exam- ple, Ni-catalyzed reactions can readily form sp2-sp3 C–C bonds, which provides methods to synthesize the types of non-planar APIs that are challenging to prepare using precious metal-catalyzed reactions. However, in general, the relative lack of mechanistic understanding about Ni-catalyzed reactions has hindered their use in the syn- thesis of APIs because it inhibits the development of improved systems and the rational design of new reactions. One difficulty in elucidating the pathway of Ni-catalyzed transformations is that NiI complexes are often invoked as intermediates but information about their reactivity is limited. In this project, novel NiI halide, alkyl, and aryl species supported by bidentate nitrogen ligands, which are proposed as intermediates in reactions including cross-coupling, cross-electrophile coupling (XEC), and metallaphotoredox based processes, will be synthesized. The ability of these NiI complexes to undergo the proposed elementary steps in catalysis will be investigated as a function of the ancillary ligand and reaction conditions using experimental and computational techniques. These studies will be complemented by experiments to probe how NiI species are formed via comproportionation between Ni0 and NiII complexes and in situ studies to elucidate the speciation of Ni catalysts during catalysis. It is expected that our fundamental investigations will lead to the design of the next generation of Ni-catalyzed reactions by providing guidelines about the reactivity of NiI complexes. Another problem with the development of Ni-catalyzed reactions is that they often involve heterogeneous reductants, which complicate mechanistic studies, create difficulties for scale up, and cannot readily be tuned to vary the reduction potential. The PI’s group has developed a series of commercially available tunable homogeneous reductants, with reduction potentials similar to Zn0. Apart from leading to improvements in practicality, the tunability of these reductants was crucial for developing novel strategies for controlling the rate of alkyl radical generation from Katritzky salts and 1° alkyl halides in Ni-catalyzed C(sp2)–C(sp3) XEC, which led to new reactivity. Here, tunable homogeneous reductants, with reduction potentials similar to Mn0, a commonly used heterogeneous reductant, will be prepared. Kinetic studies will be performed to understand the ability of the reductants to control the rates of alkyl radical formation from N-hydroxyphthalimide (NHP) esters and 1°, 2°, and 3° alkyl halides. This will be accompanied by experi- ments to identify ancillary ligands on NiII complexes that enable facile trapping of alkyl radicals, which is currently unknown. The studies on alkyl radical generation and trapping will aid in solving significant problems in C(sp2)– C(sp3) XEC, such as the use of aryl and alkyl chlorides and 3° alkyl halides as substrates. Finally, through a collaboration with Merck, the new methods will be evaluated against medicinal chemistry targets and applied to nanomole scale chemistry, which is an emerging strategy to prepare diverse libraries of bioactive compounds.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY The adaptive immune system is driven by the B Cell receptor (BCR) pathway, which triggers B cell differentiation and proliferation in response to antigen binding. Activation of the BCR generates signaling lipid phosphatidylinositol – 3,4,5 – triphosphate (PIP3) on the inner leaflet of the plasma membrane. The pleckstrin homology (PH) domain of non-receptor tyrosine kinase Bruton's Tyrosine Kinase (Btk) binds to this PIP3, triggering release of an auto-inhibited conformation and trans auto-phosphorylation. Auto- phosphorylated Btk activates downstream pathways, leading to B cell activation and proliferation. Overactive BCR signaling can lead to severe malignancies, such as chronic lymphatic leukemia and non-Hodgkin's lymphoma. The first Btk inhibitor, ibrutinib, was approved in 2013 by the Food and Drug Administration as an alternative to chemotherapy for the treatment of B cell malignancies. Successful Btk inhibition slows cancer cell proliferation by reducing the activation and binding of transcription regulator NF-κB to DNA. The most widely used Btk inhibitors are ibrutinib and second-generation derivatives, which irreversibly bind the ATP-binding pocket of the kinase domain. This pocket is highly conserved among tyrosine kinases and consequently, treatment leads to significant off-target side effects and resistance due to mutations in the binding site. These factors necessitate alternative inhibitory sites within Btk for the advancement of B-cell cancer treatment. The critical and initial step in Btk activation is its plasma membrane association through the PH domain. The PH domain represents an attractive inhibitory target as there is low sequence homology among the class. However, the lipid specificity, stoichiometry of PIP3 binding and how it regulates these assemblies, functional oligomeric states of full-length Btk, the interfaces involved, are unknown. The goal of my proposal is to determine the lipid specificity, stoichiometry, mechanism of membrane recruitment, and the membrane-associated oligomeric states of Btk. Through a quantitative understanding of these molecular events, I aim to understand how the function of Btk at the plasma membrane is regulated. I will use a workflow developed by the Gupta lab to directly detect protein-protein and protein-lipid interactions from a lipid bilayer using native mass spectrometry (nativeMS). This will allow me to detect the lipids that interact with Btk in a bilayer mimicking the lipid composition of the plasma membrane as well as the oligomeric states of membrane bound Btk. I have obtained preliminary nativeMS and vesicle association data that shows in vitro binding to PS as well as the ability to associate with bilayers in a PS-dependent manner. I will determine the functional consequences of this yet uncharacterized lipid interaction using an immortalized B Cell line. By activating the BCR within the presence of a PS scavenger, I can determine whether PS plays a role in the BCR pathway, making it a mechanism for inhibition of Btk.

NIH Research Projects · FY 2025 · 2023-09

Modified Project Summary Section Nearly one-fifth of the Unites States population resides in a rural region, and approximately one-fifth of those residents suffers from a mental illness. While these rates of mental illness are similar to urban areas, individuals living in rural regions face a disproportionate burden of negative psychiatric outcomes. Modern advances in psychiatric research have focused on using machine learning and human neuroimaging to predict diagnoses and treatment outcomes. However, recent evidence suggests that machine learning models themselves may drive differences in health outcomes through performance differences. Specifically, clinical decision-making models created in predominantly one population group may demonstrate reduced generalizability in other population groups (e.g., poorer likelihood of choosing the correct treatment if patients are rural). Given that virtually all neuroimaging ‘brain-behavior’ predictive models in psychiatry research are generated from data collected in highly populated metropolitan areas, this study will evaluate ‘brain-behavior’ models for performance differences in rural populations. It will also investigate means of eliminating these performance differences that could create further health outcome gaps in rural populations. In Aim 1, I will use neuroimaging data from 9,811 individuals in the Adolescent Brain and Cognitive Development Study to create a ‘brain-behavior’ predictive model of cognition. In Aim 2, I will evaluate this model for urban-rural performance differences and pursue strategies to reduce these differences. This study will have important implications for understanding how algorithms in healthcare drive health outcome gaps and how we can reduce these gaps by designing models that perform the same across all populations.

NIH Research Projects · FY 2026 · 2023-09

ABSTRACT Children’s exposure to intimate partner violence (IPV), often perpetrated by fathers, has been described as a gateway to other adversity, with more than a 50% co-occurrence of direct forms of child maltreatment (CM). IPV exposure can wreak havoc on children, with risk for psychosocial impairments, including posttraumatic stress disorder (PTSD), that can emerge early and cascade across development. Lacking are interventions that adequately address the complex nature of IPV in families, including fatherhood and coparenting. This gap reflects a bias towards excluding offending fathers from child-focused work and an overreliance on batterer intervention programs (BIPs), which have shown negligible effects in meta-analyses and fail to address the roots of offending behaviors in fathers. Consequently, IPV exposed children remain at risk and fathers’ personal and interpersonal functioning, including the father-child relationship, does not improve. In effect, there is an urgent need for effective interventions for fathers and their families. Fathers for Change (F4C) is a novel fatherhood-focused intervention with a dual focus on IPV and CM that focuses on identifying, understanding, and managing emotions to reduce aggression and improve partner and parent-child interactions. F4C has a growing evidence-base demonstrating significant reductions in family violence, improved father-child interactions, and in one open trial, improved child mental health. Proposed therapeutic mechanisms of F4C include reflective functioning (RF), the capacity for parents to understand their own and children’s actions as a function of underlying states and motivations, and emotion regulation (ER), the capacity to exert control over emotional states and reactions to threat. Poor RF and ER have been associated with increased family violence and stress-related psychopathology, suggesting key focal points for intervention. To date, there have been no empirical examinations of ER and RF as therapeutic change mechanisms for reducing family violence and improving father-child interactions and child mental health. Proposed is a dual-site, multi-modal examination of ER and RF in fathers (of children 4-7 y.o.) randomized to F4C (N=180) or the Duluth Model (N=180), a BIP serving as active control. In-session observational coding will assess adaptive and maladaptive ER and RF across treatment. Weekly self-ratings will assess at-home ER and RF. Aims will (1) assess efficacy of F4C compared to a standard BIP in reducing family violence and child mental health impairment, (2) identify and compare trajectories of therapeutic change targets across interventions, and (3) examine the mediating role of father’s ER and RF on child-related outcomes. This proposal will grow the evidence-base for F4C and advance our understanding of therapeutic mechanisms through which F4C exerts its effects.

NIH Research Projects · FY 2025 · 2023-08

Project Summary/Abstract The central nervous system plays a pivotal role in metabolism, within which the ventromedial hypothalamus (VMH) is crucial for proper regulation of body weight and blood glucose level. The VMH senses nutrients and circulating hormones and integrates its metabolic information to coordinate effector responses to maintain organismal homeostasis. O-linked beta-D-N-acetylglucosamine (O-GlcNAc) modification, catalyzed by O-GlcNAc transferase (OGT), is considered a cellular sensor of nutrients and hormones. Our recent studies reveal that OGT in the VMH is required for promoting adipose tissue lipolysis and maintaining energy balance by regulating the VMH neuronal activity and sympathetic innervations to white adipose tissues. Based on these findings, we hypothesize that VMH O-GlcNAc signaling determines a body weight setpoint by means of humoral and nutritional sensing and filtering lipolytic information to white adipose tissues. To test this hypothesis, we will combine gain- and loss-of- function models with state-of-the-art neural tracing and molecular profiling tools, optogenetic and electrophysiological recordings, and comprehensive physiological assessments to delineate the mechanisms by which VMH OGT regulates adipose tissue function and constrains body weight gain. Aim 1 will determine whether O-GlcNAc signaling in VMH neurons determines the setpoint of body weight. Aim 2 will delineate the neurocircuitry by which OGT-expressing VMH neurons relay lipolytic information to white adipose tissues. Aim 3 will identify how O-GlcNAc signaling regulates hormonal and nutritional sensing of VMH neurons. A detailed understanding of body weight control by hypothalamic OGT could support the development of novel therapies for obesity and co-morbidities.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY An estimated eleven million individuals are released from U.S. jails and prisons each year. Individuals with a history of incarceration have higher rates of cardiovascular events and mortality compared to the general population, especially in the weeks following release. An especially underexplored factor in the epidemiology of cardiovascular disease in this population is sleep health. Incarcerated people may have unique reasons for being sleep deficient at the individual, environmental, and institutional policy levels. They have higher rates of mood disorders and chronic pain, which amplify sleep deficiency. Exposure to extreme temperatures, noise, and light within correctional facilities and halfway houses may impact sleep. Exposure to violence and the conditions of confinement are associated with increased rates of psychosocial stress or post-traumatic stress disorder, contributing to sleep-disrupting nightmares and insomnia. The overarching aim of this study is to understand how incarceration, especially correctional and post-release social and physical environments, contribute to sleep deficiency and cardiovascular disease risk. The central hypothesis of this study is that sleep deficiency is associated with the factors in the social and physical environment during incarceration and after release. I propose a mixed methods study with the following aims: 1) Identify population-specific risk factors in the social and physical environment that contribute to sleep deficiency during and after incarceration and potential opportunities to improve sleep health; 2) Characterize using geospatial analysis the environmental factors during incarceration and post-release that may contribute to sleep deficiencies; 3) Develop and pilot instruments to assess sleep deficiency and sleep environment among people with a history of incarceration. To achieve these aims, I will leverage the existing infrastructure of an ongoing NHLBI R01 study (JUSTICE, 1R01HL137696-01A1) and the expertise of the SEICHE Center for Health and Justice, the Yale Program in Sleep Medicine, the Sleep and Health Research Program and Behavioral Sleep Medicine Program at the University of Arizona, and the National Environmental Institute of Environmental Health Sciences (NIEHS) Social and Determinants of Health Group.The proposed research and training in sleep medicine and cardiometabolic outcomes, environmental factors in sleep health, and intervention development will facilitate my transition to a career as an independent researcher in sleep health disparities among people with a history of incarceration and position my research to impact public health interventions and policies.

NIH Research Projects · FY 2025 · 2023-08

Exposing students to new opportunities, creating a sense of accomplishment, social learning through informal mentorship, and making learning fun have been found as benefits of informal STEM education. The Science & Health Partnership at Yale Center for Healthcare Simulation (YCHS) leverages advances in simulation training to promote informal STEM learning for local high school students at an academic health center. The proposed work builds on the Simulation Academy at Yale – Youth Entering Science (SAY-YES!) research team's prior innovative work developing an after-school simulation program engaging local public high school students in hands-on experiential learning using multiple technologies including high fidelity manikin simulators and virtual reality platforms to explore anatomy and physiology. During each session, a potential healthcare profession is introduced corresponding to the STEM topic. This five-year SEPA proposal seeks to develop, from the foundations of the existing SAY-YES! program, a multi-level curriculum for STEM learning, leadership, and mentoring.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY/ABSTRACT In the United States (US), the incidence of early-onset colorectal cancer (EOCRC), defined as colorectal cancer (CRC) diagnosed among individuals <50 years of age, has been increasing rapidly. Compared to late- onset CRC (i.e., CRC diagnosed among individuals ≥50 years of age), EOCRC is characterized by more aggressive pathology and distinct genetic profiles, indicating that the etiology of EOCRC may be distinct. There have been a limited number of studies examining the risk factors of EOCRC, and none have focused on early life risk factors, which could be particularly relevant to early-onset diseases. Over the past few years, the American Cancer Society and the US Preventative Services Task Force reduced the recommended age of CRC screening from 50 years to 45 years, and the COVID-19 pandemic influenced the provision of preventative services across the country. These events could impact the temporal and geographic patterns of screening for EOCRC, but no study has systematically assessed such impacts in a large sample of the US population. The proposed study aims to 1) identify early-life risk factors of EOCRC and 2) evaluate spatiotemporal patterns of EOCRC screening in the US. For the first aim, we will conduct a nested case-control study within the California birth cohort and compare the birth characteristics between 1200 EOCRC cases and 60,000 control subjects matched on year of birth. In addition, we will obtain birth address matched social, behavioral, lifestyle, and environmental factors from the Centers for Disease Control and Prevention and the Environmental Protection Agency and compare these neighborhood-level factors between the 1,200 cases and 60,000 controls. Two key methodological challenges will be addressed: a) spatial autocorrelation, the tendency of adjacent geographic units to share similar attributes; b) the modifiable areal unit problem, a biasing effect that occurs when observed associations between exposures and outcomes vary based on the scale of the chosen geographic unit. For the second aim, we will assess temporal patterns in EOCRC screening and identify factors associated with potential regional variation in screening by leveraging unique claims data from Blue Cross Blue Shield (BCBS), the largest insurance provider in the US that covers one-third of all Americans. Approximately 4 million beneficiaries aged 45-49 years are enrolled in BCBS each year, allowing us a great opportunity to assess both stool-based and structural CRC screening tests. With a rigorous design and unparalleled sample sizes, the proposed study addresses research questions with clear public health significance, is very innovative, and will probably yield a high impact on our knowledge about the etiology and screening of EOCRC – a critically understudied field according to the National Cancer Institute. In addition, by bringing together a multidisciplinary team of experts in cancer epidemiology, health services research, spatial statistics, gastrointestinal oncology, and geographic information systems science, this study will contribute to the training of a promising minority predoctoral fellow with aspirations to pursue a career in cancer epidemiology research and improve the health status for all.

NIH Research Projects · FY 2024 · 2023-08

PROJECT DESCRIPTION Campylobacter jejuni is the most common cause of food-born diarrhea in the United States. Although it is most often associated with self-limiting diarrheal disease, a small proportion of infected patients develop a more serious neuro-degenerative complication known as Guillain-Barré Syndrome. Despite many important advances in the field, still remarkably little is known about its mechanisms of pathogenicity. C. jejuni has the ability to efficiently colonize the intestine of a broad range of hosts, which allows this pathogen to gain access to the food chain thus presenting major challenges to the food industry. It has been widely observed that the attack rate of C. jejuni during well-documented point-source outbreaks or controlled human infections is under 50%, and that even under conditions of heavy exposure, only a subset of individuals developed symptomatic infections. It is well established that under physiological conditions, the resident microbiota exerts a restricting effect over bacterial enteropathogens. We therefore hypothesize that some of the documented differences in susceptibility to C. jejuni infection is due to differences in the composition of the intestinal microbiota. We intend to conduct experiments to better understand the mechanisms by which the resident intestinal microbiota restricts C. jejuni colonization. To achieve this objective we plan to leverage a germ-free chicken gut colonization model that we have recently established in our laboratory along with our long-standing expertise in the study of C. jejuni. These studies will provide the foundation for the understanding of the role of the resident microbiota in C. jejuni intestinal colonization, which may serve as the bases for the development of novel strategies to combat C. jejuni infections.

NIH Research Projects · FY 2024 · 2023-08

ABSTRACT There is an urgent need to develop novel pharmacological treatment for alcohol use disorder (AUD). Recently, psychedelic compounds have attracted great attention in treatment of different psychiatric disorders following their reported fast-acting and long-lasting effects. Preliminary evidence from observational studies, supported by animal studies, is promising for the potential therapeutic effects of psychedelic N,N-dimethyltryptamine (DMT) for AUD. Here, we propose to investigate the therapeutic potential of a single dose of intravenous DMT plus a brief course of psychotherapy (including Motivational Enhancement Therapy (MET)) on alcohol consumption in non-treatment seeking individuals with AUD, in a proof-of-concept, randomized (1:1), placebo- controlled, double-blind, parallel group, laboratory study and clinical trial. Methods: Otherwise healthy individuals with diagnosis of moderate to severe AUD (based on DSM5) will be randomized to receive a single dose of intravenous DMT or active placebo (diphenhydramine). Vitals will be closely monitored, tolerability will be measured using a Visual Analogue Scale (VAS), and DMT acute psychedelic effects will be assessed using the Mystical Experience Questionnaire (MEQ), at the end of the dosing day. Adverse events will be assessed using the Systematic Assessment for Treatment Emergent Effects (SAFTEE) interview on the dosing day and weekly follow up sessions (4 weeks). One day following drug administration, participants will attend an experimental session using Alcohol Drinking Paradigm (ADP). All participants will consume a priming dose of alcohol at the beginning of the experimental session, which will be followed by two 1-hour self-administration drinking sessions, over which participants will have a choice of consuming a total of 8 drinks or receiving $5 for each drink that is not consumed. The total number of consumed drinks is the main primary outcome. All participants will receive a brief course of psychotherapy (including MET). We will explore the effects of DMT (plus brief psychotherapy) on participants' natural alcohol consumption weekly for 4 weeks, using Timeline follow-back (TLFB), to measure the percentage of heavy drinking days, abstinent days, and total amount of alcohol consumption and categorical outcomes of abstinence, no heavy drinking and a 2-level reduction in WHO drinking risk will be compared. Hypotheses: Relative to control (diphenhydramine, IV, 25 mg plus MET), a single psychedelic dose of intravenous (IV) DMT (0.3 mg/kg) plus brief psychotherapy (including MET) in individuals with AUD will 1) be safe and well-tolerated, 2) reduce alcohol consumption measured in the laboratory using Alcohol Drinking Paradigm, the day after, and 3) reduce alcohol drinking over the following 4 weeks.

NIH Research Projects · FY 2025 · 2023-08

Summary Alzheimer's disease (AD) is a common neurodegenerative disease with progressive memory loss and cognitive decline. Early detection is critical for prevention and treatment of AD and AD related dementia (ADRD). It has been estimated that delay of the onset of dementia by even one year would reduce the prevalence of dementia by 10%. Recently, increasing evidence demonstrates that AD pathological changes can occur in sensory associated brain areas 5-10 years early before typical AD symptoms present, suggesting that they could serve as early biomarkers for AD/ADRD detection and diagnosis. Hearing is an important neural sense. Hearing loss also is a major high-risk factor for dementia. Recent studies demonstrated that visual and auditory stimulations with gamma oscillation cycles could reduce amyloid-ȕ (Aȕ expression in the brain and improve memory in AD mice. We hypothesize that hearing has a critical role in AD development and progression. However, hearing is an understudied field in AD study. Little is known about AD-induced hearing changes. Previous epidemiological studies demonstrated that AD patients could have hearing loss. However, since aged persons usually have age-related hearing loss (ARHL), it was hard to distinguish AD-induced hearing decline from ARHL in those epidemiological studies. The link to AD pathology also could not be determined and remained unclear. In this project, we will use AD mouse models to identify and characterize AD-induced functional and pathological changes in the auditory system (Aim 1). Both familial AD (fAD) and sporadic AD (sAD) mouse models will be used to increase experimental rigor. AD-induced functional changes in the auditory system will be longitudinally examined and assessed during AD development and progression. These changes will be linked to Aȕ and Tau protein expressions and genomic changes in the auditory system, which will be assessed by RNA sequencing. The AD hearing marker, thus, can be unambiguously determined. In Aim 2, we will use both AD and ARHL mouse models to further distinguish AD-induced hearing decline from ARHL. We will also define the impact of hearing loss on AD/ADRD development and progression and test whether ARHL can accelerate/exacerbate AD/ADRD development and progression. These proposed studies can improve our understanding AD pathology and the role of hearing in AD/ADRD development and progression. Such information is also critical and required for understanding the underlying mechanism for the therapeutic effect of acoustic stimulation against AD and further improving treatment and prevention.

NIH Research Projects · FY 2026 · 2023-08

Eastern Europe and Central Asia (EECA) have among the highest (5 of the top 10) incarceration rates worldwide, with staggering numbers of people who inject drugs (PWID), mostly with opioid use disorder (OUD: 82%) and HIV (PWH: 18%). The HIV epidemic in EECA is primarily concentrated in PWID, who account for 56% of new HIV infections with a high HIV prevalence among PWID (7.3-53.4%). The prisons and probation settings provide an opportunity for HIV prevention using opioid agonist therapies (OAT). The newly (past 5 years) formed probation system in these countries presents new opportunities and challenges for HIV prevention as probation does not have an international mandate to ensure human rights access to HIV prevention and treatment. This proposal builds on our existing implementation science network in several EECA countries. Specifically, first, MAT-LINK used SBIRT (screening, brief intervention, and referral to treatment) with trained researchers to pilot test a strategy to scale-up opioid agonist therapies. Second, many EECA countries have recently introduced probation, providing opportunities to ensure access to trans-institutionalized persons in probation to better align public health with public safety. Third, real-world implementation using existing infrastructure and staffing across more sites is now needed to expand OAT throughout prisons and probation. Fourth, we developed the first Project ECHO in EECA to guide OAT integration into primary care and psychiatric care into OAT settings. We have successfully used the EPIS (Explore, Prepare, Implement and Sustain) framework combined with the NIATx model of process improvement to scale-up OAT. Also using EPIS, we used Project ECHO to integrate OAT and HIV care into primary care clinics in Ukraine to teach specialized care to non-specialists, which we propose to use to teach prison doctors. NIATx is an evidence-informed facilitation strategy with documented successes in the adoption and scale-up of evidence-based practices (EBPs), like OAT. Using the EPIS framework and the NIATx approach we propose to: AIM 1: Scale-up OAT as HIV prevention in prisons and pre- trial detention in EECA and for individuals with OUD and link them to community treatment after release; and AIM 2: Scale-up OAT as HIV prevention in the newly formed probation system in EECA and for individuals with OUD and link them to OAT treatment as part of routine care to align public safety and public health. We will continue using our network to achieve these goals through collaboration with our partners in EECA. Significance is high due to the need to reduce HIV transmission in EECA where HIV, PWID and incarceration are syndemic, which is primarily concentrated in PWID and where OAT coverage is low. Innovation is high by using the NIATx model to scale-up OAT in prisons and probation, along with collaborative learning tools like Project ECHO to teach and maintain skills. Feasibility is high due to longstanding collaborations between these many collaborators and buy-in from CJS settings. Public health benefit is high due to addressing the interface of health and justice where many of the most vulnerable exist.

NIH Research Projects · FY 2025 · 2023-08

Project Summary/Abstract Adoptive cell therapy (ACT) with activated and expanded T or CAR T cells may be used to treat infections or tumors and ACT with T or CAR T regulatory cells are in clinical trials to control autoimmunity and allograft rejection. Many but not all patients benefit. The success of ACT depends upon T cell homing to relevant tissue sites. Normal circulating T effector memory or T regulatory cells can enter a tissue in response to their cognate antigen being displayed on the surface of the local microvascular ECs in a process triggered by TCR and modulated by costimulation. We hypothesize that antigen presentation by human endothelial cells (ECs) will recruit adoptively transferred in vitro expanded T or CAR T effector and regulatory cells to specific peripheral tissue sites in a process modulated by specific EC co-stimulators. In specific aim 1, we will test this hypothesis in vitro in models we have developed using endothelial cell monolayers in flow chambers to model in vivo conditions. We will expand our in vitro assys to include an examination of the effects that TCR- induced transendothelial migration (TEM) has on the T cells at the single cell level. In the case of CAR T cells, we will determine the most important costimulator receptor molecule motifs to be incorporated into the CAR for optimal TEM. We will also determine if human ECs have the capacity to cross-present or be “cross-dressed” by antigens allowing EC presentation of antigen to influence cancer immunotherapy. Finally, we will use a model we developed for studying adoptively transferred human T cell responses to synthestic microvessels assembled from human ECs, allowing genetic manipulation of the signals human ECs can provide. In aim 2 we will conduct similar experiments using T and CAR T regulatory cells. Successful completion of these aims will provide important information for extending the range of patients who may benefit from ACT.

NIH Research Projects · FY 2025 · 2023-08

Project Summary / Abstract The objective of the proposed research is to elucidate the biochemical mechanisms underlying the exquisite substrate binding and catalytic specificity of two phosphatidylinositol phosphate 5-kinases (PIP5K, PIKfyve). The PIPK family of lipid kinases include PIP5K (type 1), PIP4K (type 2) and PIKfyve (type 3), and is primarily responsible for converting phosphatidylinositol monophosphate lipids into PI(4,5)P2 and PI(3,5)P2. Despite sequence homology, these kinases are highly selective in substrate binding [PIP5K binds PI(4)P, PIP4K binds PI(5)P, and PIKfyve binds PI(3)P] and in catalytic activity [PIP5K and PIKfyve phosphorylate the C5 hydroxyl of the lipid's inositol head group, whereas PIP4K phosphorylates the C4 hydroxyl]. We and others have previously identified two structural elements within the kinase domain, the specificity loop and a conserved PIP-binding motif, that contribute to substrate selectivity, but how these two elements cooperate to confer kinase specificity remains undefined at the structural level. In aim 1, we plan crosslinking strategies to stabilize the specificity loop to facilitate co-crystallization with lipid substrates. We also plan to generate and crystallize a minimalistic catalytic core domain of PIKfyve. In aim 2, we propose genetic and chemical biological experiments to examine the role of PIKfyve in the life cycle of SARS-CoV-2. Several large-scale drug repurposing programs have identified apilimod, a PIKfyve inhibitor, as a top lead in suppressing SARS-CoV-2 replication in cell culture (a phase II clinical trial of apilimod in treating COVID-19 is ongoing at the Yale Center for Clinical Investigation). This discovery, together with earlier observations that apilimod also reduces infection by Ebola and Marburg viruses, has generated great interest in pharmacologically targeting PIKfyve. Drawing on structural and biochemical knowledge about the lipid kinase family, as well as chemical tools previously developed to target PIP4K, we have discovered a new class of potent PIKfyve inhibitors and plan to use them together with apilimod to interrogate how PIKfyve inhibition disrupts SARS-CoV-2 infection. All previously known PIKfyve inhibitors are structurally related to apilimod, and their binding mode to the lipid kinase is unknown. The new inhibitor class is significant because it not only adds confidence to the proposed involvement of PIKfyve in SARS-CoV-2 infection, but also has a known binding mode to PIPK, which should facilitate future optimization by medicinal chemistry.

NIH Research Projects · FY 2025 · 2023-08

Project Summary/Abstract Cryogenic electron-microscopy (Cryo-EM) is a new and rapidly evolving methodology for structural biology. Its success is due to advances in Direct Detector Cameras and new algorithms. This proposal seeks to develop the next generation of algorithms focused on reconstructing the structure of small molecular weight proteins, proteins with multiple conformational states, and developing algorithms that explain why resolution limits occur in Cryo-EM. The proposed research seeks to address important bottlenecks in the above problems. First, we seek to find useful statistics to detect the so-called “Einstein-from-noise” problem. This problem affects the reconstruction of small molecular weight proteins, giving catastrophically bad reconstructions. We propose to investigate this phenomenon theoretically and computationally, so that it can be detected reliably. Second, we seek to develop the next generation of algorithms for reconstructing multiple conformational states of a heterogeneous protein. In particular, we seek algorithms which are stable and which give high resolution reconstructions that can be organized in biologically meaningful ways. Finally, we seek to explore a new direction in Cryo-EM algorithms. We propose to develop a methodology which explains what causes resolution limits in a Cryo-EM reconstruction. Current methods can estimate the resolution of a reconstruction without providing any rationale for its cause. The new methodology will enable users to use data and algorithm settings more effectively. The proposed research builds on the P.I. previous NIGMS supported work on single particle reconstruction in Cryo-EM.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY Mosquitoes transmit numerous pathogens to humans by virtue of their ability to locate and bite vertebrates. Current strategies to prevent these diseases face various limitations and new approaches are needed. One possibility would be to inhibit the neurons in the mosquito brain that direct the behavior of attraction to humans and feeding on blood, but such neurons have not been identified because the tools and knowledge to do so do not exist. This proposal describes my plan to use genetic and sequencing-based approaches to identify the neurons that are required for blood-feeding behavior. I will take advantage of the fact that mosquitoes require multiple human-derived cues, particularly heat and CO2, to engorge on blood. Using gene expression changes in single cells, I will identify neurons that show altered responses specifically in the presence of both stimuli. To enable the genetic targeting of specific cell types of interest, we will create an atlas of enhancer elements that are specific to each cell type in the mosquito brain, using paired single cell RNA and ATAC-seq data. Finally, we will target key neurons for blood feeding using an enhancer driver constructs and combine them with genetic effectors to test whether the neurons are necessary and sufficient for blood feeding behavior. This proposal combines my expertise in computational genomics, transgenic mosquitoes, and behavior to achieve the goal of identifying neurons at a key node in the neural circuits for blood feeding. The identification of these neurons that drive blood feeding would be useful for understanding how they are activated and thus allow the design of more attractive mosquito traps. These neurons, if inactivated, would prevent mosquitoes from biting humans so they also could serve as a potential target to disrupt the spread of mosquito borne illness.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Over 100 million US adults have hypertension, a leading cause of mortality and morbidity, and 70% of them cannot achieve adequate blood pressure control with monotherapy alone. Although recent clinical practice guidelines suggest initiating therapy with two drugs, more than 50% of people currently treated for hypertension start with a single medication. For these patients, clinical guidelines propose adding a second antihypertensive drug for treatment escalation. The absence of head-to-head comparison in randomized controlled trials (RCTs) has limited the ability of clinical guidelines to provide evidence-based recommendations about which drug to add next for which patients. Our long-term goal is to produce real-world evidence to inform decisions about RCTs for hypertension treatment escalation and to provide the highest quality non-randomized evidence to support guideline recommendations. The overall objective in this application is to determine the comparative effectiveness and safety of the second antihypertensive agents added to monotherapy in patients with hypertension. The central hypothesis is that there is heterogeneity in the effectiveness and safety of the second antihypertensive agents, and the optimal choice depends on patient characteristics and the initial therapy. Our preliminary data demonstrate a large variation in the choice of the second agents added to monotherapy, providing ample opportunity to leverage practice variation to test our hypothesis. We will first determine the comparative effectiveness of the second antihypertensive agents added to monotherapy on major cardiovascular outcomes, such as myocardial infarction, stroke, and hospitalization for heart failure (Aim 1). We will then determine the comparative risk of the second antihypertensive agents on potential drug-related adverse events, such as acute renal failure, angioedema, gastrointestinal bleeding, and hyperkalemia (Aim 2). Finally, we will assess heterogeneity in effectiveness and safety among key patient subgroups defined by age, sex, race, and comorbidity (Aim 3). We have assembled experts in observational methods for causal inference, pharmacoepidemiology, clinical informatics, hypertension management, and implementation science, and will use real-world data from over 100 million US adults in five electronic health record (EHR) databases (i.e., Optum, Department of Veterans Affairs, Columbia, Yale, and Sentara Healthcare EHR databases). We will employ state- of-the-art observational research methods, including an active comparator new-user design, large-scale propensity score modeling, negative control outcome experiments, and empirical calibration, to emulate RCTs and to compare drug combinations. The proposed research is innovative because it will be the first study that applies massive real-world datasets and state-of-the-art observational research methods to comprehensively investigate the effectiveness and safety of the second antihypertensive agents added to monotherapy. The proposed research is significant because it provides critical evidence to inform decisions about RCTs for hypertension treatment escalation and to support guideline recommendations.

NIH Research Projects · FY 2025 · 2023-08

Project Summary and Abstract This award will support Dr. Jensen-Battaglia’s appointment as a postdoctoral fellow at Yale University’s Cancer Outcomes and Public Policy Effectiveness Research group (COPPER) and progress toward her long-term goal of developing the expertise and skills needed to become an independent cancer-focused investigator. By 2040, 73% of people in the United States (U.S.) living with cancer will be over age 65. The majority of older adults prefer to remain in their current homes as they age (‘age in place’), which is associated with health benefits. At the same time, living in deprived areas is associated with negative cancer-related outcomes ranging from delayed diagnosis and treatment to mortality, and the effects of this deprivation are intensified for those with fewer individual level resources. Housing insecurity (e.g., inadequate, unaffordable, or unstable housing) is an under-studied aspect of living environment with impacts along the cancer care continuum. Residential relocation, one sequalae of housing insecurity, has been linked to both increased utilization and disruption of healthcare. Yet there is a lack of contemporary, nationally representative data on residential relocation and its impacts among older adults with cancer. In addition, whether negative outcomes associated with relocation are related to relocation itself or deprivation of the surrounding area remains unclear. Although programs exist which support aging in place for under resourced older adults, little is known about their utilization among older adults with cancer. Identifying trends and disparities for use of programs providing long term care services in the home like Medicaid’s 1915(c) home and community-based service waivers is critical to inform future policy interventions tailored for older adults with cancer. This K00 project will address these gaps by studying adults •DJH IURP: A nationally representative dataset of U.S. older adults (Health and Retirement Study, HRS), and the Surveillance, Epidemiology and End Results program (SEER) with linked Centers for Medicare and Medicaid (CMS) claims. Using HRS data for community-dwelling older adults Dr. Jensen-Battaglia will: 1) Describe residential relocation by cancer status, 2) Compare adjusted rates of residential relocation for those with and without cancer, 3) Estimate associations between housing insecurity in the surrounding area and negative health outcomes, and 4) Explore whether these associations are stronger among those who have recently relocated. SEER data linked with Medicaid claims will allow Dr. Jensen-Battaglia to describe utilization (by state, county, and cancer characteristics) of the 1915 (c) waiver program among older adults with cancer dually eligible for Medicare and Medicaid. Dr. Jensen-Battaglia has worked closely with her mentors to develop a training plan which includes gaining expertise in the use of area-based measures and geospatial analysis, developing health policy analysis skills, and preparation to apply for future funding and a competitive, cancer- focused faculty position.

NIH Research Projects · FY 2024 · 2023-08

SUMMARY The inner cell mass (ICM) and trophectoderm (TE) are the first two cell types specified during mammalian development. TE cells support implantation and give rise to the placenta, whereas ICM cells forms the embryo and some extraembryonic tissues. Their differentiation is therefore critical for successful pregnancy. The mechanically-regulated Hippo signaling pathway is differentially activated in ICM and TE cells, driving gene expression programs that define these cell states. These programs also depend on cell type-specific chromatin landscapes. How mechanical forces regulate chromatin structure and embryonic cell fates during pre- implantation is however not fully understood. I hypothesize that mechanical forces transmitted though the cytoskeleton, regulate TE transcriptional programs by modulating both Hippo signaling and chromatin structure. In this proposal, I will test this hypothesis by defining how nuclear tension regulates Hippo signaling and chromatin organization during early embryonic differentiations. My ultimate goal is to define the mechanistic links connecting mechanical and regulatory pathways to cell and chromatin states. This work will enhance our understanding of cell fate specification, both in relationship to early embryogenesis and implantation, and more broadly. My postdoctoral work in the Giraldez lab showed that Lamin A/C is transcriptionally up-regulated in TE, compared to ICM, and that it regulates TE identify; LMNA/C depletion leads to an ICM-like transcriptional state reminiscent of Hippo pathway activation. In Aim 1 (K99), I will investigate regulation of Hippo by Lamin A/C and determine the role of mechanical sensing by cytoskeletal networks in the regulation of this signaling. In Aim 2 (K99/R00), to determine how mechanical forces regulate chromatin, I will apply advanced electron microscopy approaches to visualize nucleosome resolution chromatin structure in vivo. During the training period in the Giraldez lab, I will apply a novel labeling strategy, combined with cryo EM to characterize lamina-heterochromatin interactions. During the R00 period and beyond, I will apply these approaches to determine how compaction of the embryo and the generation of mechanical forces on the nucleus impact chromatin structure and ICM/TE fates. In Aim 3 (R00), I will use chimeric embryos and other developmental assays to examine how changes in the mechanical properties of the nucleus affects differentiation potential. I will also quantify nuclear stiffness and chromatin structure in developing mouse embryos. This work paves the way for a deeper understanding of the role of mechanical forces in regulating gene expression and cell identity. This proposal brings together training and concepts that I have acquired throughout my education and new approaches (RNA-seq and cryo-EM) that I will learn in my mentor’s (Giraldez) lab, from other scientists and specialists at Yale, and at the lab of my co- mentors, Elizabeth Villa and Berna Sozen. This proposal will complete my postdoctoral training and prepare me for my ultimate goal of running a competitive independent research program.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY/ABSTRACT Immune profiling studies continue to increase in complexity, with multi-omic designs that encompass additional dimensions such as time, tissue, and spatial profiling becoming more commonplace. Unsupervised dimensionality reduction has been a widely used and valuable approach for extracting and understanding the major sources of variation in previous studies, but popular methods such as Principal Components Analysis (PCA) and Non-negative Matrix Factorization (NMF) cannot support these increases in data complexity, nor can existing multi-omic embedding methods, which are designed for static datasets. It is critical that algorithms be developed that incorporate the complex data structures inherent in state-of-the-art immune profiling multi-omics studies that include additional dimensions (e.g., time or space) in order to capture multi-resolution components of vaccination and infection. The goal of this project is to develop algorithms based on tensor frameworks - which are extensions of matrices beyond two dimensions. Tensors naturally represent complex data without flattening on any variable, and tensor decompositions can identify multi-index patterns of variation, analogous to PCA or NMF in higher dimensions. Tensor decomposition methodology is an active area of research in the applied mathematics community, but is under-developed for application to immune profiling data, and current methods face critical challenges that prevent them from being directly applied in immunology studies. This project brings together computational immunology and applied mathematics researchers to strengthen and develop novel approaches of tensor decomposition in order to make them beneficial to the immunology community. Aim 1 will reframe a tensor decomposition problem into a regularized NMF problem, thereby allowing tools developed for matrix analysis to be used on tensor data, and furthermore will extend the new algorithm to handle multi-omics data that has a temporal or spatial component. Aim 2 will directly improve tensor decomposition approaches by developing novel metrics for tensor decomposition quality, and by extending a multi-omic embedding method into the tensor space using a novel tensor-algebra. The resulting algorithm will be able to generate components associated with data that can include both multi-omic and multi-dimensional (e.g. time, space, tissue, etc.) designs. These components can be analyzed for association with clinical features and outcomes, allowing for discovery of novel biological mechanisms. The proposed project will result in a suite of complementary algorithms that will aid the immunology community in understanding complex pathogenic and treatment/vaccination processes using the increasingly complex study designs that are becoming common to immune profiling studies.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY Respiratory infections, including SARS-CoV-2, disproportionately affect residents of correctional facilities (jails and prisons). While the Federal Bureau of Prisons and state Departments of Correction (DOCs) implemented numerous prevention strategies including social distancing protocols, vaccination campaigns, and testing programs to mitigate transmission and reduce the disease burden, overarching guidance on COVID-19 prevention within correctional facilities is limited. As a result, DOCs must develop and modify their policies based on existing evidence regarding the effectiveness of COVID-19 prevention strategies within correctional facilities. Unfortunately, the existing evidence base is limited. Specifically, the effectiveness of prevention strategies has principally been estimated in isolation (not in combination with other strategies) and while holding the variant constant. Because of this, DOCs are left with little evidence on how to implement and adapt prevention strategies in combination and under the ever-changing COVID-19 landscape. With the goal of expanding the evidence base for infectious disease prevention strategies within correctional facilities, we will estimate the effects of testing and vaccination on the burden of COVID-19 in both jails and prisons. To do so, we will develop an individual level discrete time hazard (transmission) model of SARS-CoV-2 and test the feasibility and reliability of a cutting-edge statistical causal inference approach as an alternative to transmission modeling (Aims 1 & 2). To examine the effects of testing and vaccination in combination and to identify scenarios when strategies require modification to contain spread and reduce disease burden, we will simulate waves of SARS-CoV-2 in the community and identify the strategy combinations required to prevent outbreaks within facilities using our transmission models (Aim 3). The proposed simulation approach will allow for the simulation of SARS-CoV-2 transmission and disease under known and future, theoretical scenarios. The execution of the proposed aims will strengthen the evidence available to DOCs and other policymakers and could make possible the estimation of indirect treatment effects under a causal framework within complex, nested social networks. In addition, their execution, coupled with the proposed training program comprising coursework, structured mentoring, and experiential learning, will allow Dr. Lind (the candidate) to enrich her knowledge of infectious disease transmission modeling, causal inference methods for treatment effect estimation in the presence of interference, and health disparities and infection control within a highly marginalized population, residents of correctional facilities. The candidate has established an expert mentoring and advisory team led by Dr. Albert Ko at the Epidemiology of Microbial Diseases Department at the Yale School of Public Health to enable this training, guide Dr. Lind's transition to independence during the R00 award phase and support her growth as an independently funded infectious disease epidemiologist.

NIH Research Projects · FY 2024 · 2023-08

Project Summary Pro-inflammatory cytokine TNFα is believed to be responsible for the delayed fracture healing observed in diabetes. However, there is no consensus on the effect of TNFα inhibition on the bone formation, indicating the unmet need in searching for new regents with unique features other than pure TNF inhibitors for diabetic fracture healing. Our genetic screen led to the identification of TNFR as the novel receptor of progranulin (PGRN) (Tang, et al, Science, 2011), a chondrogenic factor that has been shown to be therapeutic against autoimmune inflammatory arthritis. Type 1 diabetes is the most common autoimmune disease, characterized by chronic inflammation and elevated TNFα activity. Although TNFα activity is mediated primarily through TNFR1, we were excited to find that PGRN-stimulated bone regeneration largely depends on TNFR2. These paradoxical findings suggest that the regenerative PGRN/TNFR2 pathway plays a major role in PGRN-stimulated fracture healing. In addition, 14-3-3ε was identified as a component of TNFR2 pathway in response to PGRN stimulation. Further, we have developed an engineered protein named Atsttrin which is composed of three TNFR-binding domains of PGRN, and Atsttrin is more effective than PGRN in inflammatory arthritis. Given that elevated TNFα is believed to be responsible for delayed diabetic fracture healing, we hypothesize that PGRN and Atsttrin stimulate diabetic fracture healing through a) inhibition of TNFα/TNFR1 inflammatory and bone resorption pathway; and primarily b) recruitment of 14-3-3ε to TNFR2, followed by activation of bone regeneration pathway. The Specific Aims are: (1) To determine the role of PGRN, especially its derivative Atsttrin, in diabetic fracture healing. We will use both systemic and inducible PGRN knockout mice to determine whether knockout of PGRN delays diabetic fracture healing, and whether recombinant PGRN and Atsttrin can reverse it (SA#1A); which stage of fracture healing requires PGRN for successful completion of diabetic fracture healing (SA#1B); and whether PGRN, especially Atsttrin, has therapeutic efficacy in treating diabetic fracture (SA#1C). We will use an appropriate injectable hydrogel to locally deliver various dosages of PGRN or Atsttrin. (2) To elucidate the molecular mechanisms by which PGRN and Atsttrin stimulate diabetic fracture healing. We will determine the effects of PGRN, Atsttrin, and TNFα on chondrogenesis of diabetic bone marrow stem cells, signaling pathways, interplays and dependence on TNFR and 14-3-3ε (SA#2A); whether both TNFRs are important for mediating PGRN's role in diabetic bone healing (SA#2B); and whether the protective effects of PGRN and Atsttrin depend on 14-3-3ε by establishing diabetic fracture models with inducible 14-3-3ε[-/-] mice (SA#2C). Proposed studies will not only advance our understanding of the molecular events underlying diabetic fracture healing, but could also lead to novel therapeutic interventions for diabetic fracture healing and other conditions in which fracture healing is impaired.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Social relationships are a key component of human health and survival and impairments in social behavior have a major impact in many psychiatric conditions. Yet despite the importance of social context to health, there remains no FDA-approved medications that target social cognition and behavior. Social context is defined by the social stimuli available to an animal, is a key mediator of behavior in rodents, and impacts social choices. Yet little is known about how neuronal circuits encode social context and choice.In rodents, circuits in ACC that project to the amygdala (ACC- BLA) and Nucleus accumbens (ACC-Nac) have been shown to be necessary for different aspects of social information transfer. However, how neural activity in these regions encode social context and choice is not known. We developed a social choice paradigm in which mice choose access to a novel or a familiar/cagemate mouse. In this novel paradigm, mice consistently show preference for a social target over a novel object, but they show variable individual biases in social choice between a novel and cagemate conspecific. Recording neural activity from the ACC-BLA and ACC-Nac circuits during this behavioral paradigm will allow for a more nuanced understanding of the neural mechanisms underlying social choice. In order to better understand how activity recorded during this and traditional social behavioral paradigms we will leverage recently developed statistical models and inference algorithms to cluster nonlinear dynamical neural responses into an unspecified number of functional sub-groups called Functional Encoding Units (FEUs). We will also apply deep learning tools for behavioral analysis to engage in joint modeling of neural and behavioral data. This will enhance our ability to predict social context and enrich encoding of social behavior. Lastly, given the impact of 3,4-Methylenedioxy methamphetamine (MDMA) on social behavior and empathy and its recent clinical significance in post traumatic stress disorder, we hypothesize that MDMA paired social exposure will bias social choice in our paradigm. We will apply deep learning to behavioral analysis of our paradigm in order to test this hypothesis. Through this K-Award we will define how social stimuli are encoded in a context-specific manner within ACC-BLA and ACC-Nac circuits during social choice and how MDMA biases these social choices. In parallel, intensive mentoring, directed readings, and structured coursework will enhance my skills and toolkit in computational modeling and machine learning-based analysis of both neural and behavioral data, and behavioral pharmacology, setting the stage for independence.