New York University

NIH Research Projects · FY 2024 · 2021-12

PROJECT SUMMARY/ ABSTRACT Please see Research Strategy as per NOT-OD-23-032

NIH Research Projects · FY 2025 · 2021-09

Abstract Military veterans in the U.S. represent one of the populations most disproportionately impacted by the current opioid crisis. With historically high rates of injury and opioid analgesic use to treat chronic pain, veterans were widely prescribed opioids (often in conjunction with benzodiazepines) during the early years of the epidemic, and have been at elevated risk of opioid-related overdose and HIV/HCV infection since. Further, current data on opioid-related harms affecting veterans are likely to understate the severity of the situation, as most studies of veterans focus on the less than 50% of veterans who use VA healthcare facilities. Veterans who use opioids and are not connected to the VA healthcare system have high rates of homelessness and experience higher prevalence of comorbid substance use disorder and mental health diagnoses than their “service-connected” counterparts. Due to these vulnerabilities and the observed barriers to testing and treatment among veterans— especially substance- and mental health-related stigma, drug naiveté, and limited support networks—veterans who use opioids represent a critical target for interventions designed to mitigate overdose and HIV/HCV risk behaviors. For socially isolated veterans and veterans with limited access to healthcare, programs that work outside of formal healthcare institutions and agencies are desperately needed. This application proposes to advance scientific understanding of the most potent and efficient way to prevent opioid-related harms among veterans by achieving the following Aims: 1) Evaluate the effectiveness of a peer-delivered, community-based education, navigation and support (CENS) intervention to reduce opioid-related risk behaviors; 2) Examine factors that mediate (e.g., knowledge, self-efficacy, self-stigma) and moderate (e.g., mental health, pain/OUD severity, age) intervention effectiveness; and 3) Explore intervention participants’ and peer outreach staff perspectives on implementation as well as barriers to and facilitators of intervention effectiveness. The proposed intervention will be delivered by three veteran peer outreach workers—one dedicated to each of the three main intervention components (Education, Navigation, and Support). The study will recruit 300 veterans with opioid use disorder to participate in a randomized controlled trial. The CENS intervention will engage 150 participants in ongoing educational sessions, healthcare and treatment navigation, and social support (involving both one-on-one and group social integration protocols) designed to improve self-efficacy, reduce self-stigma, increase service and healthcare utilization, and bolster knowledge. This study stands to contribute a timely, culturally-tailored innovation to overdose and HIV/HCV prevention-as-usual that, informed by the theory of triadic influence, directly confronts the social, intrapersonal, and structural-level barriers to opioid- related risk reduction among veterans. Study findings will be of great interest to community-based and civic healthcare organizations that provide overdose and HIV/HCV risk reduction outreach, as well as to agencies committed to improving healthcare engagement among veterans.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT The disproportionate rise in nursing home (NH) use among Black and Latino persons living with dementia (PLWD) reflects the inadequacy of existing programs and policies to support aging in place for the most vulnerable. Enabling aging in the community and preventing avoidable NH placements is widely considered a priority by federal, state, and local entities along with families and older adults. Yet, it is unclear what is needed to support Black and Latino PLWD to remain in the community. While there are many plausible next steps to addressing this poorly understood problem, developing an instrument to identify which unmet needs are disproportionately driving avoidable NH placements is an important actionable step in reducing disparities among Black and Latino PLWD. The objectives of this 5-year proposed project are to 1) develop and test the feasibility, acceptability, reliability, and validity of an assessment instrument aimed at addressing the unmet needs that drive Black and Latino PLWD to experience avoidable NH placements at greater rates than their White counterparts using an exploratory sequential mixed methods research design, and 2) translate preliminary findings onto the public agenda as a policy issue. Aim 1 will use qualitative interviews to describe the unmet needs driving these placements. An integrative analysis of qualitative findings derived from Aim 1 will inform an unmet needs assessment instrument, IN-HOME (Identifying Needs to Help Older adults Maintain Everyday community living), that will be developed, refined and piloted in Aim 2 and psychometrically tested in Aim 3. Preliminary findings from this study will be used to generate actionable, evidence-based policy recommendations to address disparities in NH placements. Aligned with NIA’s strategic priorities to address disparities experienced by minority older adults with Alzheimer’s Disease or Related Dementias (ADRD), this study will form the basis for a subsequent R01 that will tailor IN-HOME for community-dwelling Black and Latino PLWD and investigate which actionable unmet needs place them at greatest risk for NH placements. The candidate, Dr. Jasmine Travers, PhD, MHS, is a gerontological nurse practitioner and assistant professor at New York University (NYU) Rory Meyers College of Nursing. The long-term goal of Dr. Travers is to build an independent research career focused on developing innovative approaches to eliminate disparities in access to and quality of person-centered long term care experienced by Black and Latino PLWD through the use of mixed methods. Under a team of exemplary mentors and scientific advisors with expertise in gerontology, ADRD, mixed-methods, policy, survey design, and psychometric evaluation, Dr. Travers will build on her preliminary work on disparities in access to community long-term care to accomplish training goals in primary research involving vulnerable PLWD, mixed methods, policy, and leadership development. NYU provides an exceptional environment for the completion of the proposed research and for Dr. Travers who has already begun to emerge as a leading disparities researcher in the field of aging and long-term care.

NIH Research Projects · FY 2024 · 2021-09

Project Summary The overarching goal of the Data Science for Social Determinants (DSSD) at New York University, Moi University and Brown University is to develop future leaders in data science who are equipped to develop and analyze data to better leverage deep and rich survey as well as internet and other digitized data sources that can help us capture information on the social determinants of health. DSSD's design will rapidly expand the local base of expertise via curriculum development, resulting in 2 PhD (4-year training) and 6 total postdoctoral (2-year) and faculty trainees (12-14 month) will train at NYU, while 8 Masters and 2 PhD trainees will commence or complete training (2-year and 4-year training, respectively) through newly developed data science tracks at Moi University. Connecting with local data science industries and organizations (IBM, Deep Learning Indaba, DataKind, AI.Kenya and Aga Khan University Nairobi and Karachi) will bring in and create intellectual meeting spaces for a wide pool and variety of talented trainees from both data science and health backgrounds, to propel and sustainably advance data science capacity in Kenyan institutions as well as the DSI-consortium.

NIH Research Projects · FY 2025 · 2021-09

During disaster crises, New York City (NYC) public transportation has been an essential service to assure that other essential workers can get to their jobs. Many transit workers are exposed to threats during disaster events, both at work and at home. Transit workers are essential workers, even though they may lack the training, experience, supplies, equipment, and supervision typically provided for traditional frontline workers (i.e., medical workers and first responders). This study, conducted in partnership with the Transport Workers Union (TWU), Local 100, is designed to: (1) evaluate the cumulative impact of multi-level interventions to date on current worker wellness; (2) develop and assess a worker-driven model of crisis management to facilitate worker well-being in the face of disasters (e.g., maintain and restore services); and (3) disseminate findings to provide input into practices and operations to protect non-healthcare essential workers during disaster events with a focus to increase their wellbeing. To achieve these aims, we propose to conduct serial cross-sectional surveys of a systematic sample of the NYC transit workforce, with the logistical assistance of TWU, representing nearly 40,000 subway and bus workers. Timing of subsequent surveys will be dynamic to capture real-time shifts and ongoing changes in strategies and practices that impact transit workers. In the first phase, we will first examine the impact of multi-level interventions already implemented by several entities, including: (a) federal, state, and local governments and agencies; (b) the Metropolitan Transit Authority (MTA), the public authority in charge of NYC Transit; and (c) TWU, which provides support and reinforcement of multilevel interventions, referrals, and social support. Guided by a new Disaster Preparedness model informed by data from our recent transit workers pilot study, the existing multilevel interventions will be evaluated to determine their impact on workers’ outcomes (e.g., well-being, psychosocial, behavioral, interpersonal relations, etc.), perceived impact of TWU interventions (e.g., reinforcement with outreach for education and support), and individual adoption of recommended practices designed to mitigate community and workplace disaster impacts. We will examine potentially moderating effects of age and occupational characteristics of the workers. Initial and subsequent survey data will inform ongoing Participatory Action Research (PAR) teams comprised of academics, workers and other key stakeholders who will formulate data-driven strategies to increase effectiveness of the multilevel interventions and further support worker resilience in the face of shifting disaster events. Results will be widely disseminated to inform policy changes suggested by study findings.

NIH Research Projects · FY 2024 · 2021-09

Project Summary Behavioral studies have shown that early life experience significantly shapes the development of brain abilities. Accordingly, if early experiences are highly unbalanced, e.g. if they occur under the influence of chronic challenges or stresses, the individual's personality will develop specific traits, including some that are associated with severe psychopathologies. Despite these extensive behavioral characterizations, very little is known about the biological mechanisms underlying learning and memory in early life, with the exception of the effects of trauma and stress. Understanding the mechanisms underlying learning and memory in early development is key for comprehending how the learning and memory systems are built and function throughout life, as well as to better elucidate the deficits associated to neurodevelopmental disabilities. One of the most important systems operating in the brain is the medial temporal lobe-dependent memory system, which processes information about episodic, spatial, contextual and social experiences. Until recently it was believed that this memory system does not function in infancy because it is developmentally immature, and only begins to be involved late in development. However, recent studies in rodent models, including our own, showed that episodic and spatial forms of learning require the function of biological mechanisms in the dorsal hippocampus (dHC), a main region, together with the medial prefrontal cortex (mPFC), of the medial temporal lobe memory system. Despite this recent progress, knowledge of the biological and system-level mechanisms of infantile, hippocampus-dependent learning and memory is lacking. To fill this knowledge gap we propose to employ rodent models of episodic and spatial learning, genetically engineered mouse models, molecular imaging technology, spatial transcriptomics and RiboTag mouse technology combined with omic analyses to pursue the following specific aims: (1) To map the distribution at a system level (dHC and mPFC) of the cellular networks activated in response to episodic learning in infancy and in memory recovery following reminders at later ages, and to test the malleability and roles of recovered infantile memories in adult behavior. (2) To comprehensively profile in situ dHC and mPFC gene expression at the level of the whole transcriptome, as well as obtain a comprehensive translatome specifically regulated in excitatory and inhibitory neurons, in response to learning in both infant and adult brains. These experiments will provide an unprecedented amount of novel information regarding the biological and system-level mechanisms underlying infantile learning and memory, as well as an invaluable source of knowledge for generating novel hypotheses regarding neurodevelopmental and adult cognitive disorders.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Hypertension (HTN) is the most prevalent modifiable risk factor for cardiovascular disease (CVD) among U.S. adults. There are well-established guidelines for managing HTN, yet HTN remains undertreated. There are a number of reasons for inadequate adoption of HTN care guidelines, including skepticism about guideline recommendations, perceptions that patients are non-adherent to medications, and lack of systems to facilitate best practices in HTN management. Team based care (TBC) has the potential to overcome these barriers. TBC is the provision of health services by at least two health providers who work collaboratively to improve chronic disease management through enhancements in quality improvement (QI) infrastructure to support care coordination, population management, self-management support, medication adherence and continuity of care. Numerous studies have demonstrated that TBC is an evidence-based strategy for improving blood pressure (BP) management and control. Despite the effectiveness of TBC, primary care practices experience significant barriers to implementing TBC care processes. This is particularly true for small-medium size independent primary care practices (SIPs), an understudied group of practices that represent the majority of outpatient care delivery but which often lack the resources and local expertise to implement care processes consistent with TBC. External practice facilitation (PF), a strategy to support implementation of evidence-based models of care into routine practice, may mitigate barriers to adoption of TBC in SIPs. Yet, there is a dearth of literature on the effect of PF on adoption of TBC in general and specifically in SIPs. The objective of this study is to compare the effectiveness of PF on the adoption of core TBC components, centered on improving BP management and outcomes in SIPs, to usual care. In this study, a PF implementation strategy will be developed through mixed methods formative assessment and input from a representative stakeholder advisory committee. This PF strategy will then be evaluated in a stepped wedge cluster randomized control trial in 90 independent primary care practices in New York City where the effect of PF on adoption of TBC (primary outcome) and blood pressure control (secondary outcome) will be compared against usual care. Given the public health burden that elevated BP presents, implementation of TBC as an evidence-based practice to improve BP control has great potential to improve CVD outcomes. This study will provide much- needed guidance on how to optimize adoption and sustainability of TBC in independent primary care setting to realize the potential of improved HTN management and addresses current gaps in prior research.

NIH Research Projects · FY 2024 · 2021-09

The CDC's Essentials for Parenting Toddlers and Preschoolers program (EFP) is a free Internet resource with the potential to break down barriers to population-wide access to scientifically-based parenting interven- tions. Our research team at NYU recently partnered with program developers and researchers at the CDC and Westat (a research firm) to evaluate EFP. We learned three important lessons from the pilot study: (1) EFP is well-liked by the parents who use it; (2) parents consume more intervention content if cued what content to use and when to use it; yet (3) the amount of content consumed, even with such guidance, is highly variable and tends to be quite limited. In short, EFP has considerable promise, but parental engagement, a major issue in the success of universal parenting interventions, remains a challenge. Thus, learning how to maximize meaningful parent engagement is the lynchpin to realizing the promise of EFP. The objective of the proposed research is to optimize EFP by identifying engagement-fo- cused intervention elements to add to EFP that enhance its effects on parenting skills. The study is based on the Multiphase Optimization Strategy (MOST) developed by Dr. Linda Collins (Co-I) and collabora- tors. The bedrock logic of MOST is that only after an intervention has been optimized to meet specific criteria, should it be evaluated in an RCT. In our view, improvements in parenting skills are the key optimization crite- ria that must be met. The study is a longitudinal factorial optimization trial in a community sample of 800 parents with 1.5- to 3-year-old children. There are 4 experimental factors; each corresponds to the presence vs. absence of an engagement-focused intervention element. This experiment enables the estimation of the indi- vidual and combined effects of each element. The specific aims follow. Aim 1: Optimize the effects of EFP on parenting skills by determining which combination of the four ex- perimental engagement-focused intervention elements results in the greatest success of EFP, as reflected in increasing parent warmth and reducing corporal punishment, overreactive and lax discipline. Aim 2: Determine the extent to which boosted meaningful parent engagement in EFP is the mechanism driving the effects of the four engagement-focused intervention elements on parenting skills, and which as- pect(s) of engagement (e.g., content consumption; behavioral skills practice) are the key mediators that trans- late the effects of the four engagement-focused intervention elements into improved parenting. Aim 3: Examine parent (e.g., change readiness; race) and child characteristics (e.g., externalizing behavior; sex) to determine if the optimal intervention package differs among subgroups. Cracking the code of providing parents with an intervention that they actually use, and that improves parenting, could have far-reaching ef- fects (e.g., improving population-level child outcomes).

NIH Research Projects · FY 2025 · 2021-09

Project Summary     Adoptive  Cell  Immunotherapy  using  patient  harvested  T  cells  engineered  against  tumor-­specific  targets  has ushered  in  a  new  therapeutic  era.  However,  the  lack  of  tumor-­specific  targets  and  T  cell  receptors  limits  the     addressable cancers. A promising approach for solid tumors uses T cell receptors (TCRs) directed against tumor-­ specific  antigens  (TSAs)  displayed  on  HLA  receptors  found  on  the  surface  of  nearly  every  cell.  Unfortunately,  HLAs are highly polymorphic genes between people. This restricts both the TCRs and TSAs to a small number  of  patients.  A  platform  that  expands  the  number  of  HLA-­restricted  TSAs  and  TCRs  will  transform  the  entire   immunotherapy pipeline.     Here,  I propose to address this unmet need by generating  programmable  Dendritic cells  (DCs)  –  professional antigen presenting cells that function to mature and activate naive T cells. Programmable DCs would permit the discovery of new TCRs, validation of TSAs, and perhaps be used as "living" vaccines to marshal a patient’s own immune system against infectious disease and  cancer. Thus far, efforts to leverage the potential of DCs have         been  limited  principally  by:  (1)  an  inability  to  produce  cells  with  HLAs  matched  to  patients;;  (2)  an  inability  to  robustly test and validate new TSAs against TCRs;; and (3) an inability to produce "off-­the-­shelf" DCs at industrial  scale.     As  a  new  innovator,  my  vision  is  to  produce  off-­the-­shelf  Dendritic  Cells  pre-­engineered  to  match  any  HLA  haplotype (even rare ones) and pre-­encoded with any combination of TSAs. Using this new platform, my group will  search  for  and  validate  "universal"  TSAs/TCRs  that  can  be  used  broadly  in  TCR-­Therapy  for  any  patient.     Specifically, we will focus on peptides expressed from regions of the genome normally epigenetically silenced,  but  re-­activated  in  tumor  cells  (such  as  endogenous  retroviruses).  To  reach  this  goal,  we  will  continue  development of technology enabling the "writing" of millions of base-­pairs of DNA in human induced pluripotent  stem cells (iPSCs). This technology allows direct customization of the large HLA locus of iPSCs in a single step;; introduction of libraries of potential TSAs;; and integration of synthetic reporter constructs (which are excisable     for safety) for enhancing the in vitro directed differentiation of iPSCs to DCs. Thus, allogeneic programmed DCs  will catalyze a wide variety of immunotherapy applications and expand access of these advanced treatments for   a greater number of patients.

NIH Research Projects · FY 2024 · 2021-08

Abstract Functional gastrointestinal disorders (FGIDs) are common, costly, and cause significant impairment that can begin in infancy. FGIDs remain poorly understood, and treatment are often ineffective, prompting a recent NIDDK workshop to conclude an urgent need for improved understanding and management paradigms. In this proposal, we test the hypothesis that serotonin reuptake inhibitor (SSRI) antidepressant exposure in pregnancy is a major contributor to FGIDs in children. This is based on the premise that SSRIs, when used by women in pregnancy, alter the fetal availability of serotonin (5-HT), which is critical for healthy nervous system development in both the brain and the gut. There are significant public health implications, as SSRI use in pregnancy is continually increasing and it is estimated that upwards of 350,000 newborns per year were exposed to an SSRI during gestation. Most of the studies to date on the long-term effects of SSRI exposures have focused on offspring brain development. In contrast, studies on the effects of maternal SSRI exposure on gut development are lacking. However, our group has compelling evidence, through both published studies and new preliminary data, suggesting associations between SSRI exposure and FGID risk, which are independent of exposure to maternal depression. We have also identified a specific microbiome profile (enterotype) that is highly linked to FGIDs, and which our preliminary results suggest, may be transferred from mother to infant where it continues to drive elevated 5-HT signaling. Thus, SSRI exposure could increase offspring FGID risk, and this risk could be mediated by microbiome changes in the mother and child. We will investigate these questions by building on an ongoing birth cohort study of 375 mother-infant dyads (“The MYRNA Study”; controls, maternal depression +/- SSRI exposure) being followed through pregnancy and the first two years of life. Our new study, which we term, “Gestational SSRI Exposures in the DevelopmenT of Functional GAStrointestinal Disorders (GETGAS)”, will leverage MYRNA infrastructure and population, while adding extensive characterization of FGID diagnoses and symptoms, and analyzing stool samples (mother in pregnancy, and infant at birth, 1 yr and 2 yr) to test the role of microbiota-driven 5-HT signaling pathways. We will do this through three specific aims: Aim 1: Confirm effects of gestational SSRI exposure on infant FGID development trajectories in the first two years. Aim 2: Test whether maternal SSRI use during pregnancy promotes a 5-HT producing microbiome that is vertically transferred to the offspring where it increases FGID risk. Aim 3: Use data driven methods to identify clusters of maternal and infant factors most predictive of FGIDs in early life. Findings from this innovative and cost-effective proposal will help disambiguate the effects of PMD, SSRI exposure, and transmission of maternal microbiota on the development of FGIDs in early life, inform clinical management of FGIDs, and provide a foundation for biomarker and therapeutic target discovery.

NIH Research Projects · FY 2025 · 2021-08

Project Summary/Abstract Our planet is inhabited by trillions of bacteria that live inside and outside of humans. The “skin”, or surface, of bacteria is called the cell envelope, and functions to separate us from them. Although some bacteria are symbionts, infection by pathogenic bacteria is still a major cause of death worldwide. While Gram-negative bacteria contain a protective outer membrane layer absent in most Gram-positives, almost all bacteria contain polymers composed of unique patterns of glycans that extend from the cell surface. Bacterial surface sugar polymers, or exo-polysaccharides, act as molecular barcodes that distinguish different strains of bacteria within a single species. Many bacterial exo-polysaccharides contain rare sugars, which are monosaccharides that are absent in other organisms, including humans. While exo-polysaccharides are necessary for host infection, we still lack an understanding of how rare sugar-containing glycan polymers are assembled, recognized, and enable survival in the host. My laboratory seeks to generate chemical and biochemical tools to study bacterial protein and glycan pathways that enable survival in different environments. Our main areas of focus are: (1) development of small molecule regulators of bacterial chaperone function; (2) manipulation of cell surface sugar patterns to selectively label and disable bacteria. This proposal focuses on the latter program, in which we identify rare saccharide subunits that are unique to Gram-negative cell surface polymers called O-antigens, and represent key epitopes that mediate interactions with hosts and susceptibility to antibiotics. Over the next five years, we will address the following questions: (1) Can we improve chemoenzymatic routes to rare sugar precursor substrates? (2) How do glycosyltransferases recognize rare sugar substrates to build O-antigens? (3) Are O-antigen glycosyltransferases regulated via protein-protein interactions? (4) What host protein structural motifs are involved in bacterial rare sugar recognition? (5) Can we identify new host proteins involved in bacterial recognition? To answer these questions, we will use a multidisciplinary approach, involving a combination of organic chemistry, chemical biology, biochemistry, microbiology and sequencing-based analyses. This work will significantly expand our understanding of cellular mechanisms underlying bacterial polysaccharide synthesis, and will teach us how humans recognize foreign sugars. Relevance to public health: In addition to providing fundamental insight into the production of bacterial factors that are important for infection, the results of this proposal will inform novel strategies to disable hard-to-treat Gram-negative infections by interference of essential host-pathogen interactions, as well as biomolecular reagents to recognize bacterial oligosaccharide structures for new diagnostics.

NIH Research Projects · FY 2025 · 2021-07

Summary Traditionally, studies of antibiotic resistance have focused on evolutionary and molecular mechanisms of resistance. However, many of our best antibiotics target the bacterial cell envelope, which is a mechanically robust, structural exoskeleton for the cell. Ultimately, these antibiotics cause cell death by weakening the envelope enough to cause explosion of the cell by the large, hydrostatic pressure within it. Despite the central mechanical importance of the cell envelope, we have little understanding of which molecules and moieties within it are critical for its load-bearing capacity. Addressing this question would transform our understanding of antibiotic resistance. A primary reason for this gap in our knowledge is the formidable challenge of applying mechanical forces to single bacterial cells while monitoring their physiology. The proposed research will address this obstacle by applying innovative, highly precise, high-throughput microfluidics and microscopy-based assays to measure the mechanical properties of two of the major cell envelope components in bacteria, the outer membrane and the cell wall. These assays will be combined with molecular and cell biological techniques, and biophysical theory, to explore an emerging paradigm within microbiology: that bacteria control antibiotic resistance by adaptively tuning the mechanical properties of their cell envelope. First, building on the recent landmark finding that the outer membrane confers antibiotic resistance to bacteria because of its mechanical strength, the dependence of outer membrane stiffness and mechanical antibiotic resistance on the fine-scale biochemical composition of the outer membrane will be systematically measured. Next, the mechanism of outer membrane vesiculation (a process underlying antibiotic resistance and pathogenesis) will be investigated by combining a theoretical mechanical model of vesiculation with novel microscopy assays to quantify vesiculation dynamics, while genetically tuning protein-protein interactions between the cell wall and outer membrane. Finally, the scope of these studies will be extended to Gram-positive bacteria by determining the dependence of antibiotic resistance on cell wall stiffness in these species, specifically focusing on the mechanical contributions of teichoic acids to resistance. Together, these studies will transform our understanding of bacterial pathogen survival and growth, and point to fresh strategies to circumvent antibiotic resistance and treat bacterial infections.

NIH Research Projects · FY 2025 · 2021-07

Project Summary Regenerative therapies offer the potential to reverse deficits arising from neurodegenerative disease, stroke, and traumatic brain injury. But the development of such treatments requires a comprehensive understanding of how to direct neurons to adopt appropriate functional properties and circuit identities. This proposal seeks to reveal fundamental principles of circuit design by identifying the permissible and predisposed molecular mechanisms evolution uses to drive changes in the courtship behaviors of drosophilids. Using a new model for comparative neurobiology that I have developed with my collaborators, I will compare homologous neurons in the pheromone processing pathways of four closely related Drosophila species. First, I will take advantage of highly stereotyped, species-specific pheromone preferences and in vivo neuroimaging to identify the sites of adaptation in pheromone processing circuits. By quantifying the courtship of each species in high resolution, I will be able to correlate differences in the pheromone preference behaviors observed between species to the changes observed in how pheromone cues are processed (Aim 1). This will elucidate the circuit motifs and dynamics that control the differential activation of an essential population of P1 interneurons that gate male entry into courtship across species. Next, to reveal the molecular underpinnings of adaptations in P1 connectivity and excitability, I will perform RNA sequencing on the P1 neurons of each species. This analysis will identify differentially expressed genes which I will test to determine how they regulate the functional properties of P1 and mate preference behaviors (Aim 2). Finally, I will assess when and how the transcription factor Fruitless–which specifies the male courtship circuitry–acts to organize the sexually dimorphic anatomy and function of P1 neurons in melanogaster males. Further, taking advantage of genetic pipelines I have built, I will use Targeted DamID to determine how changes in Fruitless target genes specify novel courtship behaviors across species (Aim 3). Under the continued mentorship of Dr. Vanessa Ruta, and supported by the substantial resources of Rockefeller University, I am well poised to complete the proposed research and shed new light on the molecular and cellular mechanisms that evolution uses to encode novel behaviors. In addition, a comprehensive career development plan, supported by my advisory committee, will ensure that I receive the conceptual, technical, and career training I require to successfully transition to independence at a top research institution.

NIH Research Projects · FY 2026 · 2021-06

PROJECT SUMMARY Anxiety and depression are increasingly recognized as disorders that are developmental in origin. While vulnerability to anxiety and depression is heightened prior to adulthood, the developmental factors that give rise to this increased risk are not well understood. Two characteristic learning and memory biases are implicated in the etiology of anxiety and depression: preferential processing of negatively valenced information and a tendency to form overly general memories and value associations. Despite their apparent clinical relevance, studies to date linking these learning and memory biases to psychiatric risk have relied largely on recollective measures that do not enable the study of how they may arise over development through value-based learning and memory encoding processes. In this proposal, we will leverage computational modeling and neuroimaging approaches to elucidate how mechanistic relations between learning computations and memory formation underlie valence and overgeneralization biases across development from childhood to adulthood. Aim 1 will characterize how valence biases in learning change over development, how they influence incidental memory for episodic details of valenced outcomes, and how they arise through neural computations. Aim 2 will characterize, across development, how generality of learned representations adapts across contexts, how the specificity of memory representations changes with time, and how neural representations support the use of multiple levels of abstraction to guide learning and memory. Aim 3 will characterize how valence and generalization biases change longitudinally with age and assess their relation to real-world autobiographical memory and clinical symptomatology. The significance of the proposed research lies in its potential to: 1) provide a theoretical account relating valence and generalization biases in value-based learning and corresponding biases in episodic and autobiographical memory; 2) elucidate the neurocognitive mechanisms underlying these biases; 3) delineate normative longitudinal developmental changes in these processes from childhood to adulthood; and 4) establish whether computational phenotypes capturing these biases predict anxious and depressive symptomatology.

NIH Research Projects · FY 2025 · 2021-06

Project Summary. Community-dwelling persons living with dementia (PLWD) are highly susceptible to avoida- ble emergency department (ED) visits and hospitalizations. Adult day service centers (ADCs) provide commu- nity-based care to a growing number of racially diverse PLWD, the majority of whom are low-income. Daily as- sessment and serial observations by an ADC’s interdisciplinary staff (which includes registered nurses, social workers, and program directors) support early detection of clinical problems in PLWD. However, when acute changes in health status occur, ADC staff who wish to provide timely notification to primary care providers (PCPs) frequently cannot do so effectively. In my prior research, I found that ADC staff relied on facsimile or voicemail message to communicate urgent information. This resulted in delayed or non-responses from PCPs and allowed minor health issues to escalate into medical emergencies. As the number of PLWD in ADCs grows, there is a critical need to strengthen communication of salient clinical information between ADCs, PCPs, and caregivers to reduce costly hospitalizations and ED visits. Mobile health (mHealth) interventions have been shown to improve communication and clinical information exchange across a variety of health care settings, but they have not been designed for ADCs. My goal in seeking a K23 award is to become an inde- pendent scientist who leads a research program that integrates care from ADCs and PCPs using mHealth in- terventions to reduce avoidable health care utilization disparities in PLWD. With support from an experienced interdisciplinary mentorship team, I will acquire training in three areas: using integrated health systems to ad- dress health care utilization disparities, developing mHealth interventions using user-centered design princi- ples, and designing and testing behavioral interventions. With the requisite training, I will execute the following specific aims: (1) identify the key domains of an mHealth application intended to support communication be- tween ADCs, PCPs, and informal caregivers regarding the health-related needs of PLWD; (2) design and test the visual layout of an mHealth application intended to support communication between ADCs, PCPs, and in- formal caregivers of PLWD; and (3) develop and examine the feasibility and acceptability of mHealth applica- tion use among ADC staff, PCPs, and informal caregivers in reducing hospitalizations and ED visits in PLWD over a 6-month period. My findings will inform a future R01 proposal to test the efficacy of an intervention using a fully operational mHealth application. This study is significant because the findings will be used to improve standards of care and reduce costly and traumatic outcomes in PLWD. It also advances legislation from the Office of the National Coordinator for Health Information Technology requiring that patients be able to access information from their medical records using their preferred smartphone application. This study is innovative because it leverages ADCs’ strengths and incorporates frontline provider perspectives to inform the develop- ment of a pragmatic user-centered mHealth application to integrate care and reduce disparities in PLWD.

NIH Research Projects · FY 2025 · 2021-05

Project Summary Repeated exposure to abuse and neglect substantially increases the likelihood of poor child and adult life outcomes. Current child welfare (CW) family preservation services (FPS), however, do not reduce maltreatment recidivism. Maternal posttraumatic stress disorder (PTSD), which has been associated with poor parent-child interaction, is a promising intervention target to reduce maltreatment. Our prior work showed that mothers receiving FPS to prevent recidivism have a high prevalence of trauma-related disorders. High rates of PTSD among these mothers suggests that treating PTSD may reduce recidivism in a significant subset of high- risk mothers. In addition to directly increasing risk of maltreatment, PTSD-related cognitive deficits may make learning parenting skills more difficult, contributing to intergenerational persistence of trauma. Hypothesis: Parenting Skills Training in Affect and Interpersonal Regulation (P-STAIR), an intervention targeting maternal PTSD-related parenting deficits, will reduce recidivism. We use a combination of STAIR to target the adverse effects of PTSD (and related depression) on parenting and an abbreviated version of PCIT to improve parenting skills. STAIR is a two-phase treatment to improve emotion regulation and interpersonal skills prior to exposure treatment. It is more effective than exposure therapy alone and increases treatment retention for child abuse-related PTSD. PCIT is a dyadic treatment that has promise for reducing recidivism in CW mothers. We added a focus on how PTSD symptoms affect parenting and parent-child interaction skills: P-STAIR. Preliminary Data: Following P-STAIR, mothers had a 7-fold lower rate of new confirmed maltreatment reports (2.7%) than NYC’s FPS population as a whole (18.6%). Maternal PTSD and depression symptoms, measured at 90-day follow-up, show that 92.3% of mothers no longer met diagnostic criteria for PTSD. Independent behavioral observations demonstrated significant reduction of negative parenting behaviors and increased positive parenting behaviors. The pilot data indicate P-STAIR is feasible. We propose to conduct a RCT to evaluate efficacy of P-STAIR compared to supportive counseling (SC). Aim 1: Compare P-STAIR to SC with respect to maternal PTSD/depression symptom reduction. Aim 2: Compare P-STAIR to SC with respect to parenting behaviors. Aim 3: Compare P-STAIR to SC with respect to maltreatment recidivism. Impact: If demonstrated efficacious, P-STAIR will reduce maltreatment recidivism among high-risk CW involved mothers, ameliorating lives of children and families and reducing maternal stigma. Our CW partners’ support letters indicate P-STAIR has potential for ready CW dissemination, thereby altering real-world clinical practice.

NIH Research Projects · FY 2026 · 2021-05

Project Summary/Abstract Over the last 15 years, several laboratories, including my laboratory, have identified multiple signaling pathways that regulate translation via the translation initiation factors eIF4E and eIF2α during protein synthesis-dependent forms of long-lasting synaptic plasticity and various memory processes in rodents, including the consolidation, reconsolidation, and extinction of auditory and contextual threat memory. These findings have generated much excitement because they demonstrate the complex biochemical regulation of translation during synaptic plasticity and memory. Despite this progress, a number of critical and unresolved questions regarding the requirement for de novo protein synthesis in memory consolidation remain unanswered. We plan to focus on auditory and contextual threat memory to determine the cell types in the amygdala and hippocampus, respectively, that require eIF4E- and eIF2α-dependent translation for memory consolidation, reconsolidation, extinction, and discrimination. We also plan to examine the cell type-specific requirement for de novo translation in memory using more complex types of behavioral paradigms, including Dysregulated translation has been shown by a number of laboratories, including my laboratory, to contribute to synaptic dysfunction and aberrant behaviors in neurodegenerative diseases such as Alzheimer’s disease (AD) and neurodevelopmental disorders such as fragile X syndrome (FXS) and autism spectrum disorder (ASD). However, using molecular approaches to dissect circuit dysfunction in these diseases/disorders has been lacking. Therefore, we plan to examine the role of cell type-specific translational dysregulation in mouse models of AD, FXS, and ASD. Moreover, we will identify the inappropriately translated mRNAs and their newly synthesized protein products using translatomic and de novo proteomic approaches that we developed to identify mRNAs/proteins that are translated/synthesized improperly in mouse models of AD and FXS. These questions will be addressed by utilizing the powerful multidisciplinary combination of new groundbreaking genetically-engineered mice and viruses, electrophysiological recordings, immuno-cytochemistry, innovative methods to measure de novo protein synthesis in vivo, cell-type specific translational profiling, and de novo proteomics. The results of these studies will provide fundamental insights into the molecular events in both excitatory and inhibitory neurons that support consolidation, reconsolidation, and extinction of memory. Moreover, these studies have the potential to provide therapeutic targets for multiple brain disorders that are associated with dysregulated translation.

NIH Research Projects · FY 2026 · 2021-04

PROJECT SUMMARY Genes play two different roles in biology, giving shape to phenotypes through developmental processes within individuals, and transmitting traits across generations through Mendelian inheritance. Evolutionary developmental geneticists work at the intersection of these roles, asking how the mechanisms that operate within individuals influence the origin, maintenance, and fate of phenotypic variation in populations. Gametogenesis and early embryonic development bridge the divide between individual and population. Development involves molecular and cellular contributions from the embryo's own zygotic genome but also from the substance of the egg and the sperm, products gametogenesis shaped by the parents’ genomes. This simultaneous regulation by multiple genomes creates distinctive inheritance properties that alter predictions about patterns of variation and divergence. To better understand how parental and zygotic genetic effects and their interactions shape variation and evolution, this project sets as its goal the characterization of genetic architectures of generation-spanning phenotypes in multiple experimental model systems, each with unique complementary features. One line of research focuses on variation in development and life history in an annelid model system, Streblospio benedicti. This species is unique in exhibiting both direct and indirect development as heritable variation, with the alternative modes representing adaptive strategies to different environmental conditions. Our population-genomic data have pinpointed tens of loci associated with the difference between two dramatically different modes of development and life-history patterns, and experimental crosses to isolate the effects of these loci allow precise reconstruction of the sequence of parental- and zygotic-effect genetic changes underlying a major evolutionary transition. These data will guide construction of improved population genetic models incorporating parental-zygotic genetic interactions. The second line of research focuses on Caenorhabditis nematodes, a longstanding experimental model for genetics. These animals exhibit sex-ratio bias and sex-associated inheritance of autosomes, both features dependent on non-Mendelian assortment during male gametogenesis. Building on the lab's extensive resources for quantitative genetic analysis in these animals, we will discover the genetic basis for variation in these traits and test hypotheses about genetic conflict between parents in the regulation of offspring sex. Regulation of chromosomal and cytoplasmic transmission, and genetic interactions between chromosomal and cytoplasmic factors, are widely shared across animals and insights from model organisms are expected to generalize to conditions of human health and disease.

NIH Research Projects · FY 2025 · 2021-03

Project Summary Oral cancer pain is more severe than all other cancers. Patients with metastatic oral cancer experience the greatest pain. Oral cancers activate neurons and produce pain; however, the effect of nociceptors on oral cancer is largely unknown. The mechanisms of reciprocal interaction between oral cancer and neurons, and how the interactions promote cancer and pain, are not known. The long-term goal is to improve management of oral cancer patients and obviate opioids by identifying components of the cancer microenvironment that are viable targets to treat cancer and oral cancer pain. The overall objectives for this application are to (i) elucidate the phenotype and distribution of transient receptor potential channel, vanilloid subfamily member (TRPV1) + neurons innervating painful and metastatic oral cancers, (ii) measure sensitization and activation of TRPV1+ neurons by mediators secreted by oral cancer, and (iii) determine the contribution of TRPV1+ neurons to oral carcinogenesis. The central hypothesis is that oral cancers release mediators, including mediators carried in extracellular vesicles (EVs) that sensitize and activate nociceptors inducing oral cancer pain. The cancer- primed nociceptors, in turn, promote cancer. The rationale for the project is that identification of components of the cancer-nerve interaction provides the opportunity to develop approaches to treat oral cancer and oral cancer pain, thereby reducing use of opioids. The central hypothesis will be tested by pursuing three specific aims: 1) Determine the type and density of innervation in oral cancers in relation to pain and metastasis; 2) Investigate sensitization of trigeminal (TG) neurons by cancer pain mediators; and, 3) Investigate cancer promotion by cancer activated and sensitized TRPV1+ neurons and evaluate the potential to stop cancer and alleviate cancer pain by antagonizing signaling via the sensory neuropeptide, calcitonin gene related peptide (CGRP). Under the first aim, neuronal innervation of the cancer will be evaluated in a retrospective patient cohort with known pain and nodal status, and testing for capsaicin (TRPV1 agonist) sensitivity and measurement of pain will be performed in prospectively enrolled oral cancer patients. For the second aim, a gene associated with pain and metastasis and miRNAs from EVs will be investigated as pain mediators. For the third aim a mouse oral carcinogenesis model will be used to investigate the impact of TRPV1 abundance on cancer incidence and phenotype, the impact of CGRP signaling on cancer promotion, and the potential for CGRP/CGRP receptor therapies for treating and preventing oral cancer and oral cancer pain. The research proposed is innovative because it is based on two new findings regarding oral cancer pain: (1) newly identified putative cancer pain mediators overexpressed in metastatic cancers from patients reporting high levels of pain, and (2) involvement of EVs in cancer induced nociceptive behavior. The proposed research is significant because these studies will lay the foundation for clinical trials to assess CGRP and CGRP receptor targeted therapies, which are FDA-approved for migraine, to treat cancer and attenuate cancer pain.

NIH Research Projects · FY 2025 · 2021-02

The overarching goal of my research program is to identify and characterize molecular mechanisms responsible for stress-induced permeabilization of the mitochondrial inner membrane. In most eukaryotic cells, mitochondria are the primary source of the energy that they provide in the form of ATP by performing oxidative phosphorylation (OXPHOS). OXPHOS is a two-step process. First, substrate oxidation by the respiratory chain results in the generation of the electrical potential on the mitochondrial inner membrane. This potential energy drives generation of ATP by the phosphorylation of ADP at the ATP synthase complex. To prevent energy dissipation and ensure that OXPHOS is efficient mitochondrial inner membrane permeability should be tightly controlled and maintained at low levels. Stress conditions associated with dysregulation of calcium and ROS homeostasis can lead to an increase in mitochondrial inner membrane permeability – a phenomenon known as Mitochondrial Permeability Transition (mPT). mPT causes dissipation of the membrane potential and loss of mitochondrial ATP-generating capacity leading to cell dysfunction and death. mPT is critically involved in a broad spectrum of diseases ranging from heart attack to neurodegeneration. Prevention of mPT is highly protective against cell death and tissue damage suggesting high therapeutics potential. However, molecular mechanisms of mPT are not well understood, and this gap in knowledge prevents mPT from being a drug target. Over the past five years, we demonstrated that mPT is a multifaceted phenomenon and depending on the disease type and stress severity, it can occur through different pathways. The central goal of our research program is to identify the link between specific molecular mechanisms of mPT and specific stress conditions. We have already established several original animal and cell disease-relevant models causing different types of mPT. In our approach, a variety of methods that measure the mPT and tissue damage at the organismal, cellular and mitochondrial levels are coupled with a number of our original electrophysiological (patch- clamp) assays that allow direct measurement of mPT at the level of mitochondrial membranes and give us a unique opportunity to dissect and characterize its multiple identities and regulation. The results of our study will provide a detailed understanding of one of the most critical events in cell death cascades and will bring an essential framework for the development of therapeutically approaches that will selectively target mPT.

NIH Research Projects · FY 2025 · 2021-02

Project Summary Sensing and processing information through signaling cascades is an essential part of cellular life. A few signaling cascades such as the MAP kinase and Hippo pathways are ubiquitous among eukaryotes yet perform different functions across organisms. Although these pathways are well-studied, how they evolve to take on new functions and adapt to new inputs remains poorly understood. The Mitotic Exit Network (MEN), a Ras-like GTPase signaling cascade and yeast homolog of the Hippo pathway, provides a unique opportunity to study this question. In the MEN, the same core signaling components operate in distinct manners under different developmental trajectories. During yeast mitosis which occurs through an asymmetric cell division called budding, the MEN is scaffolded onto the spindle pole bodies (SPB, the yeast equivalent of centrosomes) and responds to spindle position through its GTPase Tem1. During meiosis, where budding is suppressed and thus no need to sense spindle position, MEN signaling is no longer organized at the SPBs, and it is unclear whether Tem1 is still required for MEN activation and what signal it may respond to. To understand the adaptation of the MEN under distinct cellular contexts, this proposal will test the hypothesis that this adaptation is enabled partially by different activation mechanisms of the MEN kinase Cdc15, the effector kinase of Tem1, between mitosis and meiosis (Aim 1). In contrast to the drastic change in spatial organization of the MEN core components between mitosis and meiosis, the effector protein of the MEN, the phosphatase Cdc14, remains sequestered in the nucleolus prior to activation both in mitosis and meiosis. In fact, this nucleolar localization of Cdc14 is conserved from yeast to human. Sequestration of Cdc14 in the nucleolus could function either 1) to ensure tight inhibition of Cdc14’s phosphatase activity prior to activation or 2) to localize Cdc14 to dephosphorylate specific substrates in the nucleolus. To uncover the selection pressure that maintains this conserved nucleolar localization of Cdc14, this proposal will examine these two hypotheses by sequestering Cdc14 elsewhere in the cell and characterize the consequences first in yeast and then in mammalian cells (Aim 2). The experiments within both aims will be initiated during the K99 phase which also includes training of the candidate on new experimental systems such as yeast meiosis and mammalian cells, as well as the development and implementation of quantitative microscopy, proximity labeling and optogenetics. Furthermore, the candidate has assembled an outstanding mentor team to both advise her scientifically to facilitate progress of the project and prepare her for the transition to an independent investigator. Together, this proposal will create a strong foundation for an independent research career in understanding the evolution/adaptation and spatial organization of cellular signaling.

NIH Research Projects · FY 2025 · 2021-01

Oral cancer causes more prevalent and severe pain than any other cancer. Oral cancer patients suffer from severe, chronic, mechanically-induced pain; talking and eating are profoundly disrupted. Opioids are plagued with side effects and ineffective as tolerance develops; there is no satisfactory treatment for oral cancer pain. Our long-term goal is to develop cancer-targeting non-viral gene therapy to disrupt nociceptive signaling in the cancer microenvironment with minimal off-target effects and translate our approach to patients. We created two non-viral vectors with excellent transfection efficiency and no cytotoxicity: a cell-permeable peptide com- bined with a cationic lipid for DNA; and, a lipopolymer for RNA. Non-viral transfection with OPRM1 (μ-opioid receptor gene, MOR) DNA led to re-expression of MOR and partial attenuation of nociception (i.e., pain) in cancer mouse models. Protease-activated receptor-2 (PAR2) is elevated in the primary afferent neurons that innervate the cancer and drive pain. Knockdown of F2rl1 (PAR2 gene) partially attenuated nociception. While both approaches partially reduce cancer pain, our goal is elimination. Complete analgesia poses a challenge; there are multiple and redundant pain pathways, and genomic heterogeneity in oral cancer produces variations in the pathways. As a strategy to obstruct these multiple and varied pathways, we posit that a combination of OPRM1 re-expression and F2RL1 downregulation in the cancer could eliminate pain. The central hypothesis is that a non-viral vector approach to gain OPRM1 re-expression and F2RL1 downregulation within the oral can- cer will eliminate cancer pain and fully restore function. The rationale for this project is that demonstration of therapeutic efficacy of co-delivery of OPRM1 DNA and F2RL1 RNAi or CRISPR/Cas9 will yield a scientifically rigorous framework for development of non-opioid therapies that can be translated to patients. The central hy- pothesis will be tested in two specific aims: 1) Determine the feasibility and efficacy of ex vivo transfection (i.e., transfection of oral cancer cells with dual genes prior to generation of a xenograft mouse model); and, in vivo transfection (i.e., direct inoculation of genes into the oral cancer in the chemical carcinogen mouse model) with OPRM1 DNA and/or F2RL1 RNAi or CRISPR/Cas9 with the non-viral vectors to eliminate cancer-induced pain in mouse models; 2) Measure pain in oral cancer patients, analyze OPRM1 and F2RL1 expression in their can- cers relative to matched normal tissue, and analyze the correlation between dysregulation of these genes and patients’ pain. The results will give us data regarding the proportion of oral cancer patients who might benefit from co-delivery of DNA and RNA and set the stage for a clinical trial. The research proposed in this applica- tion is innovative because co-delivery of DNA and RNA into a cancer with non-viral vectors for the manage- ment of pain has not been done. The proposed research is significant because it is expected to prove strong scientific justification for the continued development and future clinical trials of novel dual gene therapy. Ulti- mately, such knowledge might lead to development of novel non-opioid therapies for cancer pain.

NIH Research Projects · FY 2025 · 2020-09

Project Summary/Abstract The proposed K01 Mentored Research Scientist Development Award will provide the applicant with advanced training and skills to launch an independent research program focused on developing and adapting culturally- sensitive interventions to reduce disparities in the psychological burden of cancer among ethnic minority cancer patients and survivors. This K01 proposal seeks to culturally-adapt and test the feasibility of a writing- based intervention, expressive helping, and its efficacy on psychological well-being, quality of life (QoL), and cancer-related fatigue (CRF) among Chinese American cancer survivors. Expressive helping integrates two distinct areas of research showing that expressive writing (i.e., a writing intervention that facilitates emotional disclosure) and helping others (e.g., support giving) improves psychological well-being among healthy and clinical populations. In expressive helping, participants write about their cancer experiences, disclosing their emotions and providing encouragement and guidance, with the knowledge that their narratives will be shared with and used as a resource for other cancer survivors. Aim 1 of the study is to conduct a qualitative study to culturally adapt the expressive helping intervention for Chinese American cancer survivors. 24 In-depth interviews and 3 focus group discussions with Chinese American cancer survivors and key informants will inform the adaptation of the expressive helping intervention by focusing on material development and recruitment, implementation, and retention strategies. In Aim 2, a mixed methods randomized controlled trial of expressive helping will be conducted to test the feasibility and preliminary efficacy on psychological well-being, QoL, and CRF outcomes (N = 108). Participants will complete follow-up outcome questionnaires at 1-, 3-, and 6-month follow-ups. A subset of the participants (n = 20) will complete post-intervention qualitative interviews to examine how writing may have influenced their psychosocial adjustment. The proposed study is well-suited to the expertise the applicant brings to the project, and well-suited to be conducted in New York City (NYC), as NYC has the largest Chinese population of any city outside of Asia. The K01 award will facilitate the applicant’s transition from conducting basic research on culture and emotion regulation with healthy Asian Americans to intervention research with minority cancer survivors. It will help build his long-term career goal in implementing evidence-based interventions to reduce cancer health disparities in the psychological burden of cancer among minority cancer survivors. As part of this award, the applicant will train in cancer survivorship, community- based participatory research (CBPR), qualitative research, and mixed methods intervention research by conducting the proposed study in collaboration with Drs. Chau Trinh-Shevrin (primary mentor), Qian Lu (co- mentor), Jennifer Leng (co-mentor), Marjorie Kagawa-Singer (consultant), and Annette Stanton (consultant).

NIH Research Projects · FY 2024 · 2020-09

A key computation that all mammals perform is determining the value of different outcomes. People and animal models evaluate outcomes as gains or losses relative to an internal reference point, likely reflecting their experience-based expectations. For example, if someone is told they will receive a particular salary at a new job, but when they start, they find that the salary is substantially less, they will view that salary (which is a net increase in wealth) as a loss relative to their reference point. Reference dependence is a consequential, ubiquitous phenomenon, driving decisions about insurance, financial products, labor, and retirement savings. The proposed work seeks to uncover how large populations of neurons represent a cognitive variable –the reference point- during value-based decision-making. This work involves complementary, synergistic interactions between experimentalists and theorists in the labs of Dr. Christine Constantinople and Dr. Cristina Savin, respectively. This proposal will develop a novel behavioral paradigm for studying reference dependence in rats, enabling application of powerful tools to monitor large-scale neural dynamics. High-throughput behavioral training will generate dozens of trained subjects for experiments in parallel. We will also develop a behavioral model to quantify key aspects of rats' behavior, including individual differences in behavior across animals (Aim 1). We will use new silicon probes with high channel counts (“Neuropixels” probes) to record from populations of neurons in dozens of rats during behavior. Recordings will be obtained from the orbitofrontal cortex (OFC), a key brain structure implicated in value-based decision-making. We will develop novel latent dynamics models that will infer the reference point directly from populations of simultaneously recorded neurons in OFC, without any knowledge of the task or rats' behavior. This model will also be able to identify aspects of neural dynamics that are common across dozens of rats, and aspects that are variable across animals, reflecting individual differences in behavior (Aim 2). Finally, we will use complementary, state-of-the-art machine-learning techniques to train recurrent neural networks (RNNs) on our behavioral and neural data. This approach will generate concrete hypotheses about the neural circuit architectures performing reference-dependent subjective valuation in our task (Aim 3).

NIH Research Projects · FY 2025 · 2020-09

PROJECT SUMMARY/ABSTRACT A fundamental but poorly understood process in eukaryotic cells is how cells structure their genomes into distinct functional domains. This project addresses this gap in knowledge by studying the centromere, a specific chromatin domain found in all eukaryotes. This stably propagated locus guides the assembly of kinetochores to ensure proper segregation of chromosomes during mitosis and meiosis. Mis-regulation of centromeres adversely affects chromosome segregation resulting in aneuploidy, a condition found in more than 90% of all cancers. Aneuploidy contributes to the development of many diseases, such as cancer and Down syndrome. The goal of this project is to understand the molecular mechanisms underlying the specification and inheritance of centromeres. In most eukaryotes, centromeres are epigenetically governed by the centromere-specific histone H3 variant, CENP-A. CENP-A partially replaces canonical histone H3 at centromeres, and provides the foundation for the assembly of kinetochores. Mislocalization of CENP-A to non-centromeric regions has a devastating impact on chromosome segregation, and has been linked to a variety of cancers. However, how CENP-A chromatin is assembled at centromeres remains poorly understood. Centromeres in most eukaryotes can be transcribed at low level, and centromere transcription has been implicated in centromere structure and function. But how centromere transcription is regulated is still largely unexplored. Interestingly, centromeres in eukaryotes are usually embedded in epigenetically distinct heterochromatin, the transcriptionally silenced chromatin domain. Heterochromatin is mainly composed of tandem DNA repeats, and has been shown to contribute to centromere formation. Nonetheless, how heterochromatin repeats are organized and the precise role of heterochromatin in centromere assembly remains elusive. In addition, centromeres and heterochromatin domains in the eukaryotic nucleus typically exhibit distinct patterns of spatial organization, which have been implicated in 3D genome architecture and regulation. But how centromeres and heterochromatin are spatially organized remains unclear. We propose to use fission yeast (Schizosaccharomyces pombe) to address these outstanding questions. Fission yeast is a simple eukaryotic model organism with many aspects of centromere regulation conserved with humans. It is particularly suited to an interdisciplinary approach that includes genetics, genomics, cytology, biochemistry, and structural biology. We propose to: 1) define the mechanisms for how centromere transcription is regulated and its role in centromere function; 2) determine how the heterochromatin repeats are organized and their contribution to CENP-A chromatin assembly; 3) identify regulatory mechanisms underlying spatial organization of centromeres and heterochromatin. Given that epigenetic regulation in fission yeast is conserved, our studies will shed light on the processes governing chromosome segregation in human cells, and contribute to a better understanding of human diseases resulting from centromere misregulation.