Washington State University

NIH Research Projects · FY 2026 · 2022-08

ESI MIRA Project Summary – Seth Rudman Project Summary A comprehensive mechanistic understanding of the process of adaptation that is predictive of outcomes is a fundamental goal in evolutionary biology and advances towards this goal have profound implications for human health. Adaptation, particularly in animal populations, is primarily studied retrospectively through the patterns it produces. Yet, decades of research have demonstrated that adaptation in natural populations occurs quickly. A growing number of field experiments, including much of my recent work, have demonstrated key insight into the process of adaptation by observing as it occurs. With advances in sequencing and bioinformatics it is now tractable to use forward-in-time experimental approaches to directly observe the process of adaptation in sexually reproducing diploid animals by studying repeated changes across independent populations, which is amongst the strongest evidence of adaptation. Combining a forward-in-time experimental approach with manipulations of aspects of genetic diversity, the selective landscape, or demography can directly test hypotheses about the genomic architecture, constraint, and pace of adaptation providing key insight into the process of rapid adaptation. I propose experiments united by an approach that leverages existing molecular and population genetics tools in Drosophila melanogaster for replicated field experiments that directly test key hypotheses about the relationship between genetic variation, adaptation, and the predictability of evolution in animal populations. The primary questions are: 1) Are genotype-phenotype relationships of complex traits predictive of the outcomes of adaptation; 2) Are large effect loci essential for rapid adaptation from standing genetic variation; 3) What is the relationship between the amount of genetic diversity and the pace and parallelism of adaptation; and 4) Does gene flow into locally adapted but declining populations promote or constrain adaptation and population persistence and does this depend on the amount of genetic diversity in the recipient population. In addition to directly answering these questions, these experiments will provide data on the pace, predictability, and importance of adaptation in shaping both genomic diversity and the persistence of populations inhabiting rapidly changing environments. The work I propose here is part of building a research program that tests fundamental questions in evolutionary biology by manipulating factors hypothesized to influence adaptation and directly observing their effects with the ultimate aim of building a mechanistic understanding of adaptation.

NIH Research Projects · FY 2024 · 2022-08

ABSTRACT Coordinated specialty care (CSC) is the standard of care for early psychosis in the US. Supported by the 5- 10% set-aside in the Community Mental Health Block Grant program, targeted at evidence-based early interventions, approximately 350 CSC programs have been implemented in 49 states across the US. Neighborhoods where individuals reside are important determinants of health and the majority of the extant work on neighborhood-level determinants and early psychosis has focused on the impact on individual-level factors such as the incidence and onset of psychosis, and duration of untreated psychosis, but very few studies have examined neighborhood-level characteristics as barrier to seeking services for early psychosis. Research has highlighted the importance of mental health services being available in proximity to an individual’s community, yet very little is known about the spatial distribution and accessibility of CSC programs in the US. There is a clear gap in what we know about the potential difficulties accessing CSC programs and whether the geographical placement and distribution of CSC programs contributes to inequities. To address this gap, our overarching goal of the proposed study is to characterize CSC programs and understand the geographical distribution and inequities in access to CSC programs. We will create a novel integrative multi-level geospatial database of CSC programs implemented throughout the US, that will include client-level data (race, ethnicity, age, gender), program-level data (geocoded location, capacity, size, setting, payment, role availability), provider-level data (race, ethnicity, professional credentials), and neighborhood-level census data (residential segregation, ethnic density, area deprivation, rural-urban continua, broadband internet subscription, public transit time). Our specific aims are to 1) Characterize the variations in facility-level characteristics of CSC programs by geographical location; 2) Examine geographic availability and accessibility of CSC programs based on neighborhood-level characteristics; and 3) Develop an interactive dashboard, using web-mapping technology, that will enable decision makers and community stakeholders to identify areas with poor access to CSC programs. The proposed research represents a novel application of advanced spatial analytics that will advance understanding on geographic inequities and accessibility of CSC programs, which can used to inform policy and the future implementation of CSC programs in high-need areas with limited access.

NIH Research Projects · FY 2025 · 2022-07

Abstract Arthropod-borne disease continues to be a significant source of morbidity and mortality worldwide. The ability of an arthropod to harbor and transmit pathogens is termed “vector competency”. Many factors influence vector competency, including how the arthropod immune system responds to the microbe. The intricacies of insect immunity have been well-studied owing the model organism, Drosophila. In contrast, comparatively little is known about tick immunity, representing a fundamental knowledge gap in vector biology. Arthropod immune processes are now increasingly recognized as being divergent across species. For example, we identified a noncanonical Immune Deficiency (IMD) pathway in ticks that limits colonization of two bacterial pathogens: Borrelia burgdorferi (Lyme disease) and Anaplasma phagocytophilum (Human Granulocytic Anaplasmosis). Ticks lack genes encoding upstream IMD pathway regulators. Therefore, the molecular and cellular events preceding the noncanonical IMD pathway in ticks deviate from the classical paradigm defined in insects. We asked in our previously funded R21 whether a specialized stress-response system termed the Unfolded Protein Response (UPR) could impact vector competency through tick immunity. Infection imparts stress on the host and, for this reason, cellular stress-responses are tightly intertwined with innate immunity. Our data shows that the UPR is induced by tick-borne bacteria and initiates the noncanonical IMD pathway in ticks. Through RNAi knockdown and pharmacological manipulation, we show that the IRE1α branch of the UPR signals through the adapter molecule TRAF2 to restrict vector colonization by A. phagocytophilum and B. burgdorferi both in vitro and in vivo. Collectively, our findings provide an explanation for how the core IMD pathway is activated independent of canonical upstream regulators. Based on these findings, our central hypothesis is that the UPR functionally regulates vector-microbe interactions through crosstalk with the IMD pathway. AIM 1 of this proposal will now investigate the role of the I. scapularis UPR on microbial growth, migration kinetics through tick tissues and/or transmission to a naïve host. AIM 2 will uncover the mechanistic linkage between the UPR and the noncanonical IMD pathway using an unbiased approach to define and characterize the signalosome during infection. Since microbial infections impart stress on host systems and cellular stress responses are well conserved across eukaryotes, we expect that the findings from this R01 will uncover novel determinants of vector competence and may have broad relevance to many arthropod-pathogen systems.

NIH Research Projects · FY 2026 · 2022-02

Anxiety disorders represent the most prevalent type of psychopathology across the lifespan. Fear/avoidance, temperament precursors of anxiety symptoms, can be observed reliably in the first year of life. Thus, develop- ment of fear, a critical component of the Research Domain Criteria (RDoC) negative valence constellation, is important in its own right, and represents an early marker of anxiety symptoms. Fearful temperament has been linked with a distinct neurophysiological signature – relative right frontal activation, detectable via electroen- cephalogram (EEG) recording. Dynamic reciprocal effects between the left and right hemispheres contribute to this asymmetry as well as associated behavioral states of fear/avoidance, and shape risk versus protection with respect to anxiety. Maternal sensitivity not only effects fear development, but also moderates links be- tween fear reactivity and anxiety symptoms. The present study addresses an important gap in research, exam- ining reciprocal effects between changes in left and right frontal activation as a vehicle for transmission of anxi- ety-related risk, along with fearful reactivity, considering maternal sensitivity as a moderator. It is hypothe- sized that (1) dominance of right hemisphere change effects on left activation growth across infancy and coupling of accelerated changes in infant fearfulness and shifts in the associated electrophysio- logical signature will confer the risk for anxiety; (2) early maternal sensitivity will serve to attenuate links between psychophysiological risk and anxiety, accentuating it later. The following specific aims are proposed: Aim 1. Infant fearfulness: Modeling behavioral and EEG changes moderated by maternal sensitivity. We will enroll mothers with infants (n=300) at three sites: Pullman, WA, Blacksburg, VA, and Jupiter, FL, re- quired to provide racial/ethnic diversity in the overall sample recruited from otherwise demographically similar communities, and because of data collection/processing demands associated with the longitudinal design. We will conduct multi-method bimonthly assessments of infant fear from 6 to 18 months, relying on a planned missingness design. Infant EEG will be recorded to measure frontal activation along with observations of ma- ternal sensitivity. Dynamic latent change effects will be modeled for behavioral and EEG indicators, providing a picture of development across infancy and setting the stage for Aim 2. Explaining anxiety onset: Contributions of latent change in fear, electrophysiology, and sensitivity. Contributions of changes in fear and related electro- physiology to the emergence of anxiety will be quantified, considering moderation of maternal sensitivity. Chil- dren will be followed until 24 months of age, obtaining questionnaire and structured interview parent-report, as well as observations of behavioral inhibition/anxious behaviors. Latent change scores for fear and frontal EEG power on the left and the right will be modeled as predictors of anxiety, with maternal sensitivity as a moderator of these links. Completion of the proposed evaluation is expected to clarify the role of fear development and maternal sensitivity in the onset of anxiety symptoms, informing preventative efforts (e.g., parent training).

NIH Research Projects · FY 2025 · 2021-09

SUMMARY (ADMINISTRATIVE CORE) Our Consortium consisting of Washington State University (WSU), the Liverpool School of Tropical Medicine (LSTM), and the Kenya Medical Research Institute (KEMRI) proposes to support KEMRI to develop and manage a highly cost-effective and sustainable KEMRI/CGHR Research Platform (KRP) in western Kenya (under “Category 1. Research Coordination and Administration” of the administrative core). The platform includes the population-based health and demographic surveillance systems, laboratories in Kisumu and Nairobi, clinical research centers in Kisumu, and offices and shared services at the Kisian campus, Kisumu. Between 2014 and 2020, KRP’s day-to-day management was changed three times to make it more efficient, transparent, affordable, and less contentious with KEMRI, CDC, and other partners. Currently, the platform is managed by KEMRI, with CDC (through its partners) contributing financially together with the other international partners who share the platform. WSU and LSTM have an excellent relationship with KEMRI and its partners and many years of administrative and technical expertise in Kenya. Following extensive discussions and agreements leading up to this joint application, WSU, LSTM, and KEMRI central leadership will support KEMRI/CGHR to implement a management plan to strengthen the Center’s capacity to assume cost-effective and sustainable management of the platform in support of their public health research in Kenya. This will include a phased introduction of financial and administrative management systems, mutually agreed systems for cost-sharing across all national and international partners sharing the facilities, close fiscal oversight by WSU, and mentorship and training. KEMRI has already taken critical steps to streamline the platform’s management and strengthen its leadership, including the appointment of three senior staff at KEMRI/CGHR, the revision of policies to enhance decision-making at the Center level, and commitment to providing its share of financial support. It also includes the rolling out of an electronic system for the management of grant and ethics applications. As part of “Category 2. Research Laboratory Administration”, we will maintain accreditation standards, quality assurance processes, and integrated biosafety and biosecurity systems to support KEMRI state-of-the-art laboratories. We will also introduce a laboratory information management system, including for effective management of samples and associated data. The KEMRI laboratories recently demonstrated their utility to the country by providing >70% of the country’s testing capacity for COVID-19 pandemic. In addition, we present evident technical expertise to lead the Research Core areas identified in the RFA, including HIV and TB, malaria, influenza, and other respiratory pathogens, zoonotic diseases, antimicrobial resistance, neglected tropical diseases, non-communicable diseases, and vaccine- preventable diseases. Experienced WSU and LSTM scientists will team up with KEMRI and CDC counterparts to design, implement, and publish research, an approach designed to train and mentor KEMRI scientists in applying and managing research funding. In the accompanying Research Core proposals, these teams describe field and laboratory activities that address each required outcome and objective of the RFA. Table 5 of this Administrative Core provides a work plan showing how each study will be implemented over the program’s 5-year duration.

NIH Research Projects · FY 2026 · 2021-08

OVERALL: ABSTRACT American Indians/Alaska Natives (AI/ANs) and Native Hawaiians and Pacific Islanders (NHPIs) are increasingly concerned about Alzheimer’s disease and related dementias, as these conditions will soon have a major impact on their communities. Although AI/ANs and NHPIs share a high prevalence of ADRD risk factors, including hypertension and type 2 diabetes, their life expectancies have improved by as much as 30 years over the last 5 decades, resulting in a tripling of their populations’ share of people ages 65+. Unfortunately, the healthcare systems that serve AI/ANs and NHPIs are unprepared for the clinical, social, and economic, burdens associated with ADRD. The Natives Engaged in Alzheimer’s Research Center will be the only P01 to include AI/ANs or Native Hawaiians since the National Institute on Aging started cataloging P01s in 1984. Based at Washington State University, with collaborators at the University of Miami, Brigham Young University, and several other major universities, as well as with AI/AN and NHPI community partners, we will create an innovative program to understand, intervene on, and mitigate ADRD-related health disparities experienced by our priority populations. Our 3 Research Projects will be supported by Administrative, Research Methods, Recruitment and Engagement, and Biospecimen Cores. The Center will also feature a network of 8 Satellite Centers led by Native researchers that encompass areas where 90% of AI/ANs and NHPIs reside. These Satellite Centers will ensure that our work and our dissemination efforts are national in scope and help to create research-ready cohorts. The Center will unite scholars across social, clinical, and behavioral sciences. Notably, 12 AI/AN or NH/PI professionals have committed to participate as Investigators or Consultants. Thus, our Specific Aims are to: 1) In a group- randomized trial, test the effectiveness of a culturally informed provider training and “dementia-friendly clinic” intervention for detection and appropriate management of AI/AN patients with ADRD and mild cognitive impairment in at least 20 urban and rural clinics serving AI/ANs; 2) In a group-randomized controlled trial, test the effectiveness of a Hula intervention for preventing cognitive decline in NHPI elders with subjective cognitive impairments or mild cognitive impairment in the Hawaiian Islands; 3) In an observational study, screen AIs in 2 remote reservation communities for obstructive sleep apnea and cognitive impairment, then conduct a randomized controlled trial to test an intervention to improve adherence to sleep apnea treatment for improving or preserving cognitive function; 4) Genotype DNA from saliva samples, catalog the frequency of genetic markers for ADRD risk, and calculate polygenic risk scores for ADRD using biospecimens from 2,700+ AI/ANs and NHPIs; and 5) Create an innovative P01 to understand, intervene on, and mitigate ADRD disparities in AI/ANs and NHPIs. Our leadership, resources, and experience offer an unprecedented opportunity to address the National Institute on Aging’s research priorities for underrepresented minorities, as articulated in the National Plan to Address Alzheimer's Disease, and to respond directly to its interest in diversifying the scientific workforce.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY MARC-WSU increases the diversity of the biomedical workforce by providing meritorious trainees from backgrounds traditionally underrepresented in STEM with the tools and experiences needed to complete their undergraduate degrees, matriculate into a graduate program, and pursue a career in biomedical research. Increasing the diversity of the biomedical workforce is the first step in investigating health disparities across America and providing innovative, effective solutions. MARC-WSU will provide a supportive environment that will inspire, enable, and empower these trainees to succeed as future leaders in biomedical research. Specifically, this program gives trainees the skills, self-efficacy, and cultural acumen to overcome systemic obstacles and flourish in the biomedical workforce. MARC-WSU will achieve these outcomes by: 1. Building community through a network of mentoring and advising relationships that support the trainees' diverse academic and career goals. 2. Increasing trainees' knowledge of the culture of biomedical research through research skills courses, mentoring, and professional development workshops. 3. Supporting the development of trainees' academic and career goals, science identity, resilience and confidence as a professional. 4. Incorporating the trainees' own career goals and values system into the research experience. 5. Enhancing trainees' self-efficacy through independent work, leadership, and peer teaching. 6. Improving trainees' oral communication, scientific writing, and scientific presentation skills. 7. Instructing trainees in safe and ethical laboratory practices, project management skills, hypothesis-driven experimental design, and rigorous data analysis. 8. Integrating knowledge from the classroom and the research experience to develop scientific understanding and critical thinking skills. 9. Enriching the research experience with attendance and presentations at local and national symposiums. 10. Expanding the number of underrepresented trainees applying to and matriculating into biomedical PhD programs. The MARC-WSU program will measure success using individual student progress in critical thinking, writing, and speaking skills, completion of advanced coursework in the sciences and engineering, the cultivation and practice of research skills, and the matriculation of MARC-WSU trainees into doctoral-level programs in the biomedical sciences, with successful careers beyond the completion of the PhD. The progression of MARC- WSU trainees into biomedical careers represents a pathway for increased diversity of the biomedical workforce, and consequently for improved wellbeing of underserved communities across the nation.

NIH Research Projects · FY 2025 · 2021-05

PROJECT SUMMARY/ABSTRACT Understanding how to induce skin regeneration instead of scarring will have broad implications clinically and cosmetically because; they inhibit mobility, are painful, and are a source of psychological trauma. In the United States, treatment for burn victims alone amounts to $7.5 billion annually, the total market for scar treatment is estimated to be around $12 billion. A lack of understanding of the role of fibroblast heterogeneity inhibits progress in developing novel treatments for clinical conditions associated with scarring, such as, scleroderma, diabetic ulcers, psoriasis and fibrotic keloid scarring. Consequently, understanding how to regenerate skin has the potential to impact anyone who undergoes surgery, but also individuals with clinical conditions associated with scarring. Adult skin wounds heal with scars, but embryonic skin can regenerate without scarring. However, not knowing how to safely transfer the regeneration abilities of embryonic skin to adult skin remains a gap in knowledge. Thus, the long term goal of our research is to establish methods to induce fibroblasts in adult skin to have the same regenerative potential as their embryonic skin counterparts, without altering development and homeostasis. The objective of this application is to induce adult fibroblasts with embryonic transcription factors to reprogram skin to support regeneration and reverse aging. Embryonic and Neonatal skin have a transient Papillary Fibroblasts (PFs) that are lost during skin maturation and aging. Neonatal PFs reside beneath the epidermis, support skin regeneration, and express the canonical Wnt transcription factor, Lef1. A hallmark of skin aging is the degradation of PFs and the loss of Lef1 expression. Our central hypothesis is that Lef1 is the master regulator of Neonatal PF function, which induces scar-less skin regeneration and will restore PF identity in aged skin. Guided by our exciting preliminary data we will use novel transgenic mouse models and next generation sequencing technologies to investigate this hypothesis by the following specific aims: In aim 1 we will test the hypothesis that Lef1 is the master regulator of Neonatal PF function by tissue specific induction in adult skin. Here we will use wounding experiments and chamber grafting assays to test if adult scarring fibroblasts can be transformed to be regenerative. In aim 2, we will define the downstream pathways that Lef1 regulates to specify Neonatal PF functions. To do this we will perform ChIPSeq and Single-Cell-RNA-Seq with our regenerative transgenic model systems. Finally, in aim 3 we will determine if Lef1 can safely restore Neonatal PF identity and function in aged skin. We will analyze “old” transgenic mice with reprogrammed skin and test their ability to regenerate skin and inhibit “aging”. This proposal is innovative because of our novel approach of manipulating embryonic transcription factors in fibroblasts in vivo. The proposed research is significant because understanding how to safely regulate fibroblast lineages will lead to new molecular and cellular targets to regenerate organs, while also providing an important knowledge base for other clinical conditions associated with dysfunctional fibroblasts such as; psoriasis, scleroderma, and arthritis.

NIH Research Projects · FY 2025 · 2021-05

PROJECT SUMMARY/ABSTRACT Alzheimer’s disease and related dementias (ADRD) affects 5.5 million Americans. The number of people in the US with ADRD is expected to increase to 16 million by 2050 unless preventive interventions and effective treatments are developed. Despite advances in ADRD research among non-Hispanic Whites, little is known about the prevalence and risk factors for ADRD in American Indian and Alaska Native (AI/AN) populations. This is in part due to low participation of AI/ANs in ADRD research. The National Institute on Aging and the Alzheimer’s Association have issued calls for an increase in minority participation in clinical trials. However, success has been evasive in efforts to recruit AI/AN populations into ADRD clinical studies. The internet is a primary method for providing information about ADRD and recruiting participants into clinical studies. AI/ANs frequently use the internet for health information but also report that the websites they access lack cultural sensitivity. To increase AI/AN participation in ADRD research, it is critical to understand how AI/ANs consume and process online information. This information can be used to create effective communication and recruitment material for ADRD clinical studies. In this K01, we will apply a multi-method approach to assess how AI/ANs process and consume online material. Using this information, we will create culturally tailored online content to educate AI/ANs about ADRD to promote their enrollment into ADRD research. Our Specific Aims are to: 1) conduct psychophysiology testing and semi-structured interviews to characterize visual website search patterns, objectively measured cognitive and emotional responses, knowledge about ADRD and research, and preferences for accessing health information online; 2) create a culturally tailored educational ADRD website for AI/ANs that offers research opportunities and conduct a pilot study to demonstrate feasibility of a future randomized controlled trial; and 3) complete a rigorous training plan that will position the applicant as an independent investigator and leader in communication with underserved populations, especially AI/ANs. This innovative application provides an opportunity to discover critical information about how AI/ANs process information and answers the National Institute on Aging’s call for research to better understand effective strategies for recruiting minorities into ADRD research and communicating health messages that are appropriate for diverse populations.

NIH Research Projects · FY 2025 · 2021-05

PROJECT SUMMARY / ABSTRACT Advances in machine learning and low-cost, wearable sensors offer a practical method for understanding, assessing, and intervening for Alzheimer's Disease and Related Dementias (ADRDs) in everyday spaces. We propose to create a Behaviorome research program that will create ground-breaking methods for building health-predictive models from wearable sensor data by mapping patterns of behavior using machine learning and pervasive computing technologies. This program will create innovative multidisciplinary ideas to address NIH ADRD Milestone 11.c, Embed wearable technologies/pervasive computing in existing and new clinical research. Our research program builds on a history of interdisciplinary research contributions in areas including human behavior modeling from longitudinal sensor data and design of novel assessment and intervention mechanisms. We propose to design and validate methods for mapping a human behaviorome “in the wild”, automatically assessing cognitive and functional health from behavior markers, scaling technologies through machine learning, linking health and behavior with their influences, and closing the loop with automated interventions. Similarly, our mentoring program builds on experience training students and early- career investigators to become leaders in the field of gerontechnology. We will recruit and train graduate students and early-stage researchers, including those from underrepresented groups, to grow an institutional multidisciplinary Behaviorome research program and to establish new research programs that contribute to the targeted Milestone. We will scale the impact of mentoring by establishing a webinar series and creating youtube videos that highlight and explain breakthroughs in the design and application of Behaviorome research. Results of this program will include scripts and templates to construct a behaviorome with resource- limited wearable devices, scale data and models to large diverse populations, integrate data with multiple information sources (e.g., genetics), automate health assessment and intervention, and create understandable explanations of data and models. These will contribute to existing clinical studies such as the clinician-in-the- loop smart home, digital memory notebook, and pervasive computing measures of functional performance. Furthermore, they will lead to new clinical studies that formalize connections between health and its influences, exploration of the impact of ethnicity and the built environment on health, and the design of ADRD interventions for medication adherence, task prompting, and negative interaction de-escalation. The proposed contributions are significant because they will provide insights on detecting and assessing ADRDs within a person's everyday environment using wearable sensing and pervasive computing methods that have not been investigated in prior work. Additionally, the mentoring steps will pave the way for a new generation of researchers to offer improved methods of addressing the need to understand, assess, and intervene for ADRDs in everyday settings, thereby improving quality of life and reducing health care costs.

NIH Research Projects · FY 2025 · 2021-05

PROJECT SUMMARY/ABSTRACT Together, tobacco and alcohol kill more than half a million people in the United States every year, making co- addiction to these substances the leading cause of preventable death. We propose a contingency management (CM) paradigm based on point-of-care urine tests that measure ethyl glucuronide. Theoretically framed within the Addiction Neuroclinical Assessment (ANA) with hypothesized mechanisms in core domains (i.e., incentive salience, negative emotionality and cognitive function), and proceeding from a sound and innovative scientific premise, we will, for the first time, target smoking and alcohol use by implementing an evidence-based behavioral treatment (CM) for alcohol among participants who have initiated frontline pharmacotherapy (varenicline; VC) for smoking cessation. This project seeks to replicate, harness and extend our previous findings from secondary analyses and pilot studies, which demonstrate that applying CM to target the use of alcohol can produce non- targeted reductions in smoking. We will do so by offering CM for alcohol use to smokers who are simultaneously initiating smoking cessation pharmacotherapy. We also seek to identify the most potent ANA-based mediators (e.g., incentive salience given that CM’s focus is on alternative reinforcement) of behavior change in response to treatment. Lastly, we will examine the extent to which biologic sex and baseline alcohol use and cigarette smoking interacts with CM to produce different levels of reduced alcohol use and cigarette smoking. Patients will take part in a 2-arm randomized controlled trial to evaluate the ability of a CM intervention to reduce alcohol use and cigarette smoking. After completing a 2-week induction period, patients will be randomized into 2 trial arms for a 12-week treatment period: 1) a CM intervention group that receives smoking cessation treatment as usual (TAU), which includes varenicline and counseling, and 2) a non-contingent (NC) control group that also receives TAU. Our Specific Aims are to: 1) Determine if CM+TAU is more effective than NC+TAU for reducing alcohol use and cigarette smoking. We hypothesize that CM will yield lower rates of biochemically-verified alcohol use and cigarette smoking, concurrent biochemically-verified alcohol use and smoking, and self-reported drinks per day, cigarettes per day, and heavy drinking days during the 12-week treatment and 6-month follow-up periods. 3) Identify the most potent ANA-based mediators of treatment response across the treatment groups. We hypothesize that higher executive dysfunction, negative emotionality, and alcohol- and smoking-related incentive salience will be associated with higher levels of alcohol use and cigarette smoking across treatments and over time; 3) Determine if biological sex or baseline severity of alcohol use and smoking interacts with treatment assignment to produce differential changes in our primary and secondary outcomes. We hypothesize that biological sex will significantly interact with treatment to produce differential outcomes, and that those with less severity of alcohol use and smoking at baseline will experience better outcomes. This investigation will support future studies that can identify personalized treatment for different sub-groups of smokers with an AUD.

NIH Research Projects · FY 2025 · 2021-05

Project Summary/Abstract Prostate cancer (PC) affects 1 in 9 men and causes nearly 30,000 yearly deaths in the United States. Understanding the PC tumor microenvironment (TME) is essential for optimizing cancer prevention and care. Unlike highly mutable tumor cells, non-cancerous stromal cells in the TME, consisting primarily of cancer associated fibroblasts (CAFs), are a genetically stable and attractive therapeutic target in PC, with reduced risk of acquired resistance from genetic changes. Our long-term goals are to elucidate the molecular mechanisms governing stromal support of PC growth and progression, and identify potential druggable therapeutic targets in the stromal compartment to disrupt tumor-stromal interactions. We recently found that monoamine oxidase B (MAOB), a mitochondrial oxidative enzyme responsible for degrading monoamine neurotransmitters and dietary amines, is highly induced in PC stromal cells compared to normal prostate stromal cells in patient samples, PTEN-knockout (KO) transgenic mice and primary cultures of patient-derived CAFs. Stromal MAOB expression further increases during disease progression toward castration resistance and neuroendocrine differentiation, compared to hormone-naïve disease. Our epidemiological studies revealed that men taking MAOB inhibitors for neurological disorders such as depression tend to have a lower incidence of PC. Functional studies showed that MAOB ablation in prostate stromal cells profoundly suppressed co-cultured PC cell proliferation/invasion and co-inoculated xenograft/allograft prostate tumor growth in mice. Mechanistically, gene profiling, bioinformatics and phenotypic analyses indicate that stromal MAOB heightens cellular reactive oxygen species (ROS) levels and chemotaxis/chemokine secretion, particularly CXCL13. Based on these new findings, we hypothesize that the elevated expression of MAOB in PC stromal cells promotes adjacent epithelial PC development and progression, and that targeting MAOB and its downstream effectors in stromal cells is an effective strategy to treat PC. In Aim 1, we will determine the functional role of MAOB in stromal activation and its contribution to tumor growth and progression in tissue recombinant xenograft immunocompromised mice and MAOB-KO immunocompetent mice. In Aim 2, we will investigate the molecular mechanism by which MAOB mediates tumor-stromal communication in PC, specifically dissecting how Twist1 cooperates with TGFβ1/Smad3/4 by ROS to activate CXCL13 and how the CXCL13/CXCR5 paracrine axis impacts PC cell behaviors and associated novel signaling pathways in the context of MAOB. We will also establish the relevance of our mechanistic findings in a large collection of human PC samples and assess correlations with disease status. In Aim 3, we will evaluate the efficacy of MAOB inhibitors for treating PC, including castration-resistant PC, in xenograft and syngeneic mouse prostate tumor models. These studies have tremendous biological, pathological and clinical implications for the future application of clinical MAOB inhibitors and development of TME-targeted therapies to complement existing tumor cell-centric PC therapies.

NIH Research Projects · FY 2025 · 2021-04

Abstract: Exposure to solar ultraviolet (UV) light creates DNA damage that induces high levels of somatic mutations in human skin cancers like melanoma. UV light most commonly causes cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts (6-4PPs) at dipyrimidine sequences (i.e., TT, TC, CT, and CC). Deamination of cytidines in these lesions or mutagenic bypass leads to their frequent conversion to mutations. As a result, UV- exposed cells and skin cancers are dominated by C to T and CC to TT substitutions in dipyrimidine sequences that collectively constitute a UV mutation signature. Surprisingly, many driver mutations that contribute to melanoma progression have sequence context and substitution characteristics that do not conform to the canonical UV mutation signature. For example, the most common melanoma driver mutation, BRAF V600E, involves a T to A substitution in a GTG sequence context. This difference in mutation characteristics between most UV-induced mutations and melanoma driver mutations has led to the hypothesis that UV light induces melanomagenesis by mechanisms other than the induction of mutations. However, we have recently provided experimental evidence of mutations caused by rare, non-canonical UV lesions in whole genome sequenced yeast and bioinformatics evidence of similar lesions in human clinical skin cancers. These lesions are likely bulky photoproducts formed at TA, CA, and AC dinucleotides. Strikingly, the mutations associated with these atypical UV photoproducts have identical characteristics to many recurrent driver mutations in melanoma, suggesting that these rare lesions may play a significant role in causing skin cancer. The objective of this proposal is to better define the characteristics of atypical UV photoproducts and their contribution to cancer progression. In Aim I, we will utilize CPD and 6-4PP photolyases expressed in yeast to assign UV-induced mutation classes to their corresponding lesion types as well as assess the ability of physiological UVB light to induce mutation classes associated with atypical UV photoproducts in yeast and human cells. In Aim II, we will analyze the genome-wide distribution of atypical TA photoproducts using a novel high throughput sequencing method, called UVDE-seq. We will also characterize the formation of putative CA and AC photoproducts and their contribution to oncogenic BRAF mutations. Finally, Aim III will identify DNA polymerases involved in the error-free and error-prone bypass of TA and AC photoproducts. Successful completion of these aims will provide new insights into the molecular causes of skin cancer, and thereby define a new paradigm of UV mutagenesis that could potentially explain the epidemiological association of acute UV exposure with increased melanoma incidence.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY/ABSTRACT Endometriosis is estimated to affect 10% of reproductive age women. It results in considerable morbidity with chronic and debilitating pain, which substantially affect the quality of life of women and their families. Because it is an estrogen-dependent disorder, hormonal therapies are available for the medical treatment of endometriosis. However, these hormonal treatments along with laparoscopic surgery are often of limited efficacy with high recurrence rates, frequent side effects, additional costs, and potential morbidity. Thus, a critical need exists to develop new and effective therapies for endometriosis targeting biologically important mechanisms that underlie pathophysiology of this disease. Endometriosis is known as a chronic inflammatory disease. Aberrant inflammatory dysfunction contributes to development and progression of the disease. We have recently determined a small molecule, niclosamide (Niclo) that could serve as a potential new effective, non-hormonal, fertility-sparing option for the treatment of endometriosis. We have demonstrated that Niclo reduces growth and progression of endometriosis-like lesions (ELL) via inflammatory signaling using a mouse model of endometriosis. Our studies show that large peritoneal MΦ (LPM) are increased in the peritoneal fluid (PF) of ELL mice and invaded into the ELL. Elevated LPM populations in the PF are reduced by Niclo. Niclo also inhibits aberrant inflammation established in the PF, ELL, pelvic organs (uterus and vagina) and dorsal root ganglion (DRG), as well as MΦ infiltration, vascularization and innervation in the ELL. Therefore, we hypothesize that understanding the complex chronic inflammatory mechanisms associated with endometriosis is crucial to develop a new therapeutic strategy and provides the rationale for targeting immune dysfunction. The objective of this application is to test this hypothesis by examining: 1) how loss of LPM impacts pathophysiology of endometriosis, 2) how ELL induction alters the functionally heterogenic population of ELL and peritoneal exudate cells, and how their inflammatory dysfunction is inhibited by Niclo and 3) how inhibitory interactions from Niclo correlate with pain-related symptomology. Niclo is an efficacious Food and Drug Administration-approved drug for the treatment of helminthosis in humans that has been used for decades. Thus, drug re-purposing of Niclo could result in a rapidly-distributable, fertility-sparing and effective non- hormonal therapy that has fewer side effects than current treatments.

NIH Research Projects · FY 2025 · 2021-03

PROJECT SUMMARY This NIH ORIP K01 award application describes a 5-year training plan designed to allow me to gain additional skill and knowledge so that I can transition to an independent R01-funded tenure track research scientist. In carrying out the proposed research and career development plan, I will add to my scientific repertoire and acquire expertise in intestinal stem cell biology, microbiome, and microfluidic organ-on-chip technology. Using this newly acquired expertise, I will establish a scientific niche that will set me apart from my mentors and pave the way to a robust, extramurally funded research program. With the support of my mentoring team, I have designed a robust research program that leverages my extensive expertise with comparative gastroenterology and molecular biology. Specifically, Aim 1 will demonstrate molecular and genetic alterations affecting canine IBD following the development and validation of a patient-specific IBD model that can quantitatively assess the cellular and molecular signature of host-microbiome crosstalk. Aim 2. will allow mapping of the microbial signature and epithelial integrity in response to the host-microbiome intercellular crosstalk by utilizing single-cell level multi-omics (especially genomics and transcriptomics) and RNA in situ hybridization. Consistent with the ORIP’s mission statements promoting veterinary scientists to employ their expertise in comparative medicine to investigate human diseases, my research will allow me to use my expertise in comparative gastroenterology as well as in primary stem cell culture to investigate alterations in intestinal homeostasis relevant to Inflammatory Bowel Disease. Also, as ORIP supports animal modeling of human diseases, I will be using the dog as a spontaneous animal model to investigate the effect of gut microbiota in the intestinal epithelium given their genetic and physiological similarity to humans. The results generated in this proposal have direct implications for human diseases, since they will provide new insights into genetic and transcriptomic alterations initiating or maintaining the chronic inflammation in the gut. Such findings can be applied to various chronic conditions that have been epidemiologically associated with microbiome dysbiosis and disturbances of intestinal health (i.e., Colorectal Cancer, Diabetes Mellitus, and Alzheimer’s Disease, to name a few). This knowledge may be applied to understand disease development and novel therapies aimed at modifying intestinal homeostasis via perturbation of epithelium-microbiome-immune axis in the intestine. In summary, the training goals and career development activities proposed in this application will promote my successful transition into independent research directions.

NIH Research Projects · FY 2026 · 2021-02

Among load-bearing implants, total hip arthroplasty (THA) is probably the most clinically successful intervention. CoCrMo alloy, a wear resistant material of choice, is typically used in femoral heads for THAs. In vivo life of THAs are often reduced due to debris generation, and Co and Cr metal ion release from modular junctions. Management of taper corrosion from trunnions of CoCrMo head and Ti6Al4V stems remain a serious challenge today. Taper corrosion happens primarily due to mechanically assisted crevice corrosion (MACC) along with fretting and galvanic corrosion, and leads to adverse local tissue reactions (ALTR), an immune- mediated biological reaction due to elevated Co and Cr ions. ALTR has profound influence on bone, leading to implant failure, which can result in early revision surgery. Co and Cr ions can also cause other symptoms such as deafness, blindness, and interstitial cell damage resulting in impaired renal functioning. Our application is focused on self-lubricating and self-healing calcium phosphate (CaP) reinforced Ti- or CoCrMo-alloys to minimize bio-tribocorrosion in applications such as trunnions in modular taper interlocks in THAs. CoCrMo-CaP composite will be designed to minimize Co and Cr ion release compared to pure CoCrMo alloy; while Ti alloy- CaP composites will be designed to completely eliminate the release of Co and Cr ions due to corrosion or wear degradation. The objective of this proposed research is to test our central hypothesis that CaP based solid lubricants in Ti or CoCrMo alloys will form an in situ film at the contact surface to minimize bio- tribo-corrosion and reduce metal ion release. The rationale is that once we understand the mechanisms of tribofilm formation and its influence on bio-tribo-corrosion, we can design implants with reduced metal ion release possibility in vivo. Our preliminary data show in situ tribofilm formation with CaP reinforcement in Ti6Al4V or CoCrMo alloys during in vitro bio-tribo- corrosion studies. Presence of tribofilm lowered wear induced damage and minimized metal ion release in vitro. We have three Specific Aims for the proposed program – (1) to understand tribocorrosion mechanism and tribofilm formation in CaP reinforced Ti-alloy matrix composites, and measure their in vitro biological response; (2) to understand tribocorrosion mechanism and tribofilm formation in CaP reinforced CoCrMo composites, and measure their in vitro biological response, and (3) to measure in vivo biological properties of CaP added Ti or CoCrMo alloys.

NIH Research Projects · FY 2025 · 2021-01

Microcystin-LR (MCLR) is the most potent and abundant cyanotoxin produced by freshwater blue-green algae. MCLR exposure is associated with nonalcoholic fatty liver disease (NAFLD) and hepatocellular carcinoma (HCC). NAFLD has quickly risen to become the most common chronic liver disease in many countries, and HCC is the most common type of primary liver cancer. My K99/R00 data indicate MCLR drives progression of nonalcoholic steatohepatitis (NASH), an advanced form of NAFLD, to a more severe burnt out phenotype (increased fibrosis and reduced steatosis). The burnt out stage is an important pathogenic step towards irreversible cirrhosis and HCC. Preliminary data in Sprague Dawley rats indicate that MCLR-elicited burnt out NASH persists 4 weeks after MCLR exposure, in contrast to the control diet group that returned to a baseline phenotype. In addition, MCLR persistently dysregulated matrisome genes (related to fibrosis) and carcinogenesis genes (de-differentiation/stem cell markers) in NASH. Based on these data we hypothesize MCLR-mediated fibrotic and carcinogenic reprogramming of the liver is enhanced in NASH and promotes the development of cirrhosis and HCC. This hypothesis will be tested in two aims. Aim 1 will determine the role of the protein phosphatase 2A (PP2A)-associated pathways in MCLR-mediated stellate cell activation and hepatocyte de-differentiation. Considering the liver is the primary target for MCLR, there is surprising paucity of research investigating these pathways and none have examined the cell type specific effects of PP2A. STUDY 1.1 will define the contribution of specific PP2A-associated pathways responsible for MCLR-elicited hepatocyte and stellate effects using MCLR and PP2 modulators in human HepaRG hepatocytes and human LX2 stellate cells co-cultured as 3D spheroids in static culture or microfluidics. STUDY 1.2 will test the hypothesis that hepatocytes are the primary MCLR target and stellate cells are activated by hepatocyte damage using HepaRG and LX2 cells cultured individually or in combination in transwell plates. STUDY 1.3 will define the effects of PP2A- pathway modulators on MCLR- and NASH diet-elicited liver pathology in rats fed either a control or a NASH diet. Aim 2 will determine the mechanisms and long-term effects of MCLR-elicited fibrotic and carcinogenic reprogramming on cirrhosis and HCC development in healthy versus NASH. STUDY 2.1 will mimic important aspects of the original MCLR tolerable daily intake study including the dose (40 µg/kg), interval (daily), and duration (3 months) in the context of a tumor initiator (diethylnitrosamine) and a NASH diet. These mechanistic studies will be complemented with phosphoproteomics and single nuclei transcriptomics analyses, potentially providing druggable targets. This research will have sustained impact because it will be the first to comprehensively assess MCLR-elicited cirrhosis and HCC in pre-existing liver disease. By mimicking the TDI study, this research may indicate a lower TDI is required for people with pre-existing liver disease.

NIH Research Projects · FY 2025 · 2020-12

PROJECT SUMMARY Stress is a pervasive aspect of daily life and a significant risk factor for a host of mental illnesses, including major depression. In the brain, chronic stress causes adaptations in the mesolimbic dopamine system that increase vulnerability for developing depression and depression-related behaviors in clinical populations and preclinical animal models, respectively. One area of the brain that has gained attention as of late is the lateral habenula (LHb), in part because of its ability to tightly constrain dopamine activity. Notably, the LHb is hyperactive in individuals suffering from major depression, while restoring normal activity in this area has emerged as a viable therapeutic strategy in treatment-resistant patients. Although we still do not know how chronic stress leads to LHb dysfunction, one intriguing possibility is through stress-induced alterations in the endogenous cannabinoid (ECB) system. The primary role of the ECB system in the brain is to provide activity- dependent, on-demand negative feedback, which helps to maintain synaptic homeostasis. Our data indicate that chronic stress augments ECB signaling in the LHb, while local activation of this system elicits a passive- despair-like coping strategy, impairs behavioral flexibility in an attentional set-shifting task, and decreases the firing rate of dopamine neurons located in the ventral tegmental area. However, the precise role of the ECB system in the LHb and the mechanisms by which this system modulates stress-related behaviors has yet to be formally evaluated. In the current proposal, we will fill this important gap in knowledge by systematically examining how the ECB system modulates LHb function and identifying whether chronic stress-induced alterations in this system are necessary to produce deficits in dopamine cell firing and the expression of depression-related behaviors. In Aim 1, we will perform site-specific pharmacological manipulations of the ECB system in tandem with in vivo electrophysiology recordings of dopamine cell activity in freely behaving rats to uncover how stress-induced alterations in LHb ECB signaling may contribute to deficits in behavioral flexibility. In Aim 2, we will use ex vivo electrophysiology combined with retrograde labeling of LHb projections to identify the role of the ECB system in modulating excitatory and inhibitory LHb inputs and examine how chronic stress alters ECB control of synaptic strength at projectionally defined LHb synapses. In Aim 3, we will use a combinatorial viral approach to determine effects of acute, circuit-specific activation of LHb neurons on stress coping and behavioral flexibility, and test whether chronic, long-term LHb activation recapitulates the behavioral effects of chronic stress in an ECB-dependent manner. Broadly stated, the proposed research will fill a significant gap in the field by identifying how the ECB system regulates the activity of a key circuit that has been implicated in various domains of mental health, and the neurophysiological and behavioral consequences of stress-induced alterations in this system. Moreover, this work will pave the way for future studies exploring the involvement of this system in brain function and disease.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY / ABSTRACT The world's population is aging and the increasing number of older adults with Alzheimer's disease and related dementias (ADRDs) is a challenge our society must address. While the future of healthcare availability and quality of services seems uncertain, at the same time advances in pervasive computing and intelligent embedded systems provides innovative strategies to meet these needs. One particular need which technology can help address is assessment and assistance with a person's functional performance. The long-term goal of this work is to develop technologies that will improve the independent functioning and quality of life of individuals with functional limitations (particularly individuals with ADRDs) and reduce their reliance on caregivers. The primary objective of this application is to develop a multi-modal sensor-based approach to automate functional health assessment and assistance with everyday activities. Building on our prior collaborative work, our approach will be to collect and fuse multi-modal functional performance data from ambient sensors, mobile sensors, free text, and assessment apps (Aim 1). This fused “human behaviorome” will provide a basis, together with observation-based ground truth, for automated functional assessment and validation of each component technology, including the use of compensatory strategies, through in-person observation and through video recording of typical daily activities and strategies (Aim 2). Finally, using iterative, user-centered assessment of prompt-based assistance, we will evaluate the ability of activity segmentation and forecasting techniques to provide automated support for activity initiation and accurate completion of everyday activities (Aim 3). The proposed contributions are significant because they will provide insights on functional health revealed within a person's everyday environment that have not been investigated in prior work. The results can also help to extend functional independence through real-time assistance, while the outcomes can assist family planning, provision of care, and design of real-world and lab-based measures of functional performance. This work is important because of the increasing number of older individuals experiencing cognitive and functional limitations due to chronic health conditions. Furthermore, they address the need for individuals to remain functionally independent as long as possible in their own homes, thereby improving quality of life and reducing health care costs.

NIH Research Projects · FY 2024 · 2020-09

Project Summary This Mentored Research Scientist Development (K01) application will provide the candidate with the necessary skills to become an independent investigator focused on the development and implementation of alcohol misuse interventions for American Indian (AI) young adults. The activities in the K01 include utilizing community-based participatory research to culturally re-center a Contingency Management alcohol intervention that combines technology-based engagement strategies and behavioral economic theoretical models to increase retention of AI 18-29 year olds. To support the achievement of career and research goals the candidate has carefully assembled a team of experts in the areas of culturally tailoring substance use disorder treatments among AI people, community-based participatory research, behavioral economics, Contingency Management and technology delivered alcohol interventions among young adults (Drs. Kamilla Venner; Nina Wallerstein; James Murphy; Matthew Pearson; Michael McDonell and J. Scott Tonigan). The candidate’s training goals include: 1) Acquire the content knowledge to develop culturally grounded alcohol use disorder interventions and apply behavioral economic theoretical models with AI communities; 2) Harness technology in the delivery of alcohol use disorder interventions; 3) Strengthen expertise in mixed, methods longitudinal and advance statistical analyses; and 4) Grant writing. In partnership with a rural reservation community, the candidate will implement an enhanced Contingency Management (CM) intervention for at risk drinkers (AUDIT > 8) aged18-29 year old, with a focus on enhancing treatment attendance and engagement in substance free activities. This objective will be achieved in two phases, with Phase I consisting of establishing a Community Advisory Board that will guide the research and assist with conducting four focus groups (n = 8 per group) and twenty semi-structured interviews with young adults, providers and cultural leaders to identify the form and structure of a culturally centered approach to treatment and engagement in CM and substance-free activities. Phase II will involve a 12-week two-group RCT (n = 60) that compares standard CM with the re-centered CM plus. Fixed assessments will occur at baseline and at 4, 8, and 12-weeks and the enhanced CM condition will employ mobile devices to assess substance free activities weeks 10-12. Study aims will address (1) whether rewarding treatment attendance and alcohol abstinence increases retention and abstinence, and (2) assess daily community recovery alternatives and engagement in substance-free activities. As an exploratory analysis, we will also assess secondary outcomes to determine responders and non-responders to the planned intervention. Collectively, career development and research activities are tightly integrated and mutually supportive thus offering an ideal platform for Dr. Hirchak to become an independent investigator.

NIH Research Projects · FY 2026 · 2020-09

PROJECT SUMMARY WSU’s MIRA is an ESTEEMED Program that increases the diversity of the bioengineering workforce by providing meritorious students from underrepresented and/or disadvantaged backgrounds with the tools and experience needed for success in college and as applicants to research-based graduate programs in bioengineering. Increasing the diversity of the bioengineering workforce is the first step in investigating research participation disparities across America and providing innovative, effective solutions. By adapting and adopting evidence-based practices and interventions in novel ways, MIRA provides a supportive environment that helps develop underrepresented undergraduate scholars’ academic and research potential. Specifically, this program gives students the skills, strategies, and cultural acumen to overcome systematic obstacles and flourish in the bioengineering workforce. MIRA will achieve these outcomes by: 1. Building community through mentoring, motivation, monitoring, and advising. 2. Creating an environment of support and motivation that will socially integrate students into the WSU STEM community. 3. Facilitating academic integration with academic advising and an enriched curriculum in areas of professional development, science communication, and professional writing. 4. Increasing student research expertise and confidence as a scientist and professional. 5. Integrating students into the culture of biomedical research with research preparation courses, seminars, and workshops. 6. Enhancing the undergraduate research experience with attendance and presentations at local and national symposiums. 7. Increasing the number of underrepresented minority students engaged in a productive research experience. 8. Transitioning students to 3rd and 4th year research experiences in the WSU Honors program. The WSU MIRA program will measure success using individual student progress in critical thinking, writing, and speaking skills, completion of advanced coursework in the sciences and engineering, the cultivation and practice of research skills, and the matriculation of MIRA scholars into doctoral-level programs in bioengineering. The successful progression of MIRA scholars into bioengineering careers represents a pathway for increased diversity of the bioengineering workforce, and consequently for improved wellbeing of underserved communities and acceleration of scientific innovation.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY/ABSTRACT Alcohol use disorder (AUD) is an alarming public health problem that costs the US more than $249 billion annually. AUDs leads to a devastatingly diverse set of negative health outcomes, including significantly increased risk of hypertension, stroke, pancreatitis, liver cirrhosis, Alzheimer’s disease, multiple cancers, and multiple types of cognitive dysfunction. Disulfiram, naltrexone, and acamprosate are the only FDA-approved medications for the treatment of AUD and are only modestly effective. Zonisamide (ZON) is an anticonvulsant medication with GABAnergic, glutamatergic and monoaminergic effects which has shown significant promise in animal and human laboratory settings, and small clinical trials for the treatment of AUD. ZON could represent a critical new tool for clinicians, but previous studies have been limited by a couple of key factors (e.g., no objective evidence recent alcohol consumption, no objective evidence of medication adherence) that this study is designed to overcome. Theoretically framed within the Addiction Neuroclinical Assessment with hypothesized mechanisms in core domains (i.e., incentive salience, negative emotionality and cognitive function), and proceeding from a sound scientific premise we will target reduced alcohol use among AUD patients in primary care using a carefully designed combination of contingency management (CM, or incentives in exchange for objectively verified evidence of behavior) delivered at thrice-weekly visits during the first 4- weeks of ZON treatment to jumpstart reductions in drinking. For the remaining 8-weeks of ZON, CM will be delivered at weekly visits in exchange for evidence of 100% or higher medication adherence. The goal of this study is to complete a double-blind, placebo-controlled RCT to evaluate the ability of ZON to significantly decrease alcohol use among treatment-seeking AUD adults. Individuals will be randomized into 1 of 2 trial arms that will receive: 1) ZON plus standard treatment (ST), which includes Take Control bibliotherapy modules and CM through the 12-week treatment period (ZON+ST), or 2) Matched placebo plus ST (PLO+ST). The primary outcome will be biochemically-verified alcohol use during the treatment period. We will use a 2-week, single- blind placebo induction period wherein participants will be required to demonstrate a 75% medication adherence rate before continuing into the 12-week treatment period. Follow-up will occur at 1, 6 and 12-months. Our Specific Aims are to: 1) Determine if ZON+ST is more effective than PLO+ST for reducing biochemically- verified alcohol use and other alcohol use outcomes; 2) Identify ANA-based mediators of treatment response across the treatment groups; 3) Determine a.) if biological sex, baseline depression and baseline severity of alcohol use interacts with treatment assignment to produce differential changes in our primary and secondary outcomes; and b.) whether groups differ on adverse events or medication adherence. This project focuses on the efficacy of a promising pharmacotherapy for AUDs using a double-blind placebo-controlled design that will rigorously measure alcohol use and medication adherence.

NIH Research Projects · FY 2024 · 2020-08

Abstract This revised R01 application aims to use natural medicinal compounds (NMCs) to enhance bioactivity of synthetic bone grafts. The delivery of NMCs in synthetic bone grafts, such as calcium phosphate (CaP) scaffolds, is critical to modulate bone cell-materials interactions to improve in vivo osteogenesis and angiogenesis. Using NMCs, we propose to investigate osteoinductive, 3D-Printed (3DP) CaP bone grafts without using any growth factors (GF) or proteins towards applications in orthopedics and dentistry. However, there still exists a clinical need to innovate osteoinductive synthetic resorbable biomaterials that will promote angiogenesis, exhibiting biological properties similar to autografts. We propose to use NMCs such as curcumin from turmeric, acemannan from aloe vera, and essential nutrients such as Vitamin D3 to improve osseointegration in CaPs instead of using GFs. In our preliminary studies, we have seen promising results showing angiogenesis and enhanced osteogenesis due to the presence of NMCs. We have found that curcumin can enhance osteogenesis and angiogenesis of 3DP CaP scaffolds. From our previous work, we have also established that addition of dopants such as Mg2+, Si4+, Zn2+ and Ag+ in CaP can improve mechanical properties, enhance osteogenesis and angiogenesis while improving antimicrobial response in vivo. Combining our exciting preliminary results on NMCs and established data on dopant chemistry, we propose to demonstrate next generation of synthetic bone grafts without any GF or proteins for applications in dentistry and orthopedics, which is the premise of this application. The objective of this research is to test our central hypothesis that NMC-loaded doped CaP porous 3D Printed (3DP) scaffolds will enhance osteogenic and angiogenic properties in vivo. The rationale is that once we understand the required dose for different NMCs, their release kinetics, and the mechanism of bone cell- materials interactions modulation, the key knowledge gaps, we can design GF free synthetic bone grafts similar to autografts in clinical applications. Our long-range goal is to demonstrate clinically relevant patient matched, osteogenic and angiogenic doped CaP or CaP– polymer scaffolds with NMCs that will substitute autologous bone for repair, replacement, and augmentation in orthopedics and dentistry. To achieve our research objectives, we propose two Specific Aims. Aim 1 is focused on understanding the influence of NMCs on enhanced bioactivity of porous CaP ceramic, and ceramic-polymer composite scaffolds in vitro. Aim 2 is focused on measuring osteogenesis and angiogenesis in vivo using NMC loaded porous CaP ceramic and ceramic-polymer composite scaffolds. If successful, NMCs can also work with other materials for dental and orthopedic applications.

NIH Research Projects · FY 2025 · 2020-07

Project Summary/Abstract Clinical studies indicate that the aversive somatic states experienced during opioid withdrawal predict drug craving and increase the risk of relapse. However, little is known about the neural circuitry mediating these aversive states, or how that circuitry interacts with known components of the neural circuitry mediating relapse to opioid seeking. Recently, the paraventricular thalamic (PVT) projection to the nucleus accumbens (NAc) was implicated in opioid withdrawal symptoms in mice exposed to repeated non-contingent morphine. However, it is unknown whether withdrawal from heroin self-administration engages the PVT➝NAc pathway to drive relapse by precipitating an aversive and/or painful state. To fill this gap in knowledge, during the K99 mentored phase of this proposal, I will receive training in optogenetics and brain slice electrophysiology to manipulate neural circuits implicated in heroin withdrawal and relapse. I will use optogenetics and chemogenetics to determine whether this pathway is necessary and sufficient to drive aversion or hyperalgesia during heroin withdrawal, as well as relapse to heroin seeking. I will validate the functionality of the optogenetic and chemogenetic approaches in vitro using brain slice electrophysiology. My preliminary data indicate that activating the PVT➝NAc pathway is sufficient to drive aversion and heroin seeking during acute and extended withdrawal after abstinence from heroin self-administration, but not after extinction training. The PL➝NAc pathway has been proposed as a “final common pathway” to drug seeking and it has been shown to drive heroin seeking after extinction training. The extinction procedure may thus engage the prefrontal cortex to diminish the role of the PVT➝NAc pathway in heroin seeking. A central hypothesis of this proposal is that the prelimbic cortex (PL) drives heroin seeking during acute opioid withdrawal and after abstinence through an indirect PL➝PVT➝NAc pathway, but through a direct PL➝NAc pathway after extinction training. During the R00 independent phase of this proposal I will test this hypothesis using chemogenetics to inhibit the PL➝PVT pathway to attenuate heroin withdrawal-induced hyperalgesia and relapse. One means by which this shift in circuits may occur, depending on withdrawal modality, is by shifting glutamatergic drive to the same population of NAc neurons that drive heroin seeking. To assess this, I will investigate whether a subset of NAc neurons receives convergent inputs from both the PL and PVT using slice electrophysiology. Another means by which this circuitry may evolve to drive heroin seeking is through changes in synaptic strength within PL➝PVT➝NAc pathway. I will thus investigate synaptic strength (measured as AMPA/NMDA ratio) within this circuit. Next, I will test the ability of a long-term depression protocol (LTD) in vitro to normalize identified changes in synaptic strength, and then apply the same protocol in vivo to reduce heroin seeking and hyperalgesia during withdrawal from heroin self-administration. These experiments will reveal the role of PVT circuits in heroin withdrawal and how these circuits interface with known relapse circuitry to control heroin seeking.

NIH Research Projects · FY 2024 · 2020-07

ABSTRACT Due to an alarming rise in opioid use among the general population that is mirrored in pregnant women, Neonatal Abstinence Syndrome (NAS) rates have increased in the US from 2004 to 2014. Most newborns experiencing NAS require non-pharmacologic care, which entails, most importantly, maternal involvement with her newborn. Facilitating postpartum maternal-newborn involvement is critical in preventing further adverse maternal-newborn outcomes. To achieve positive maternal-newborn involvement, mothers need to learn effective caregiving NAS strategies while they are pregnant. Surprisingly, current obstetrical practice standards for high risk pregnant women do not address this pressing need, in part because no interventions exist to prepare future mothers for the challenges of caring for their newborns at risk for NAS. To address this critical gap, I propose to adapt an existing mobile NAS tool for clinician training and decision support, for high-risk pregnant women and assess its usability, acceptability, and feasibility in a small randomized controlled analog trial. First, I will conduct semi- structured interviews with a panel of neonatology experts, NAS care providers, and mothers with NAS-affected babies to gather their recommendations on management of NAS and explore their perspectives on the care of these newborns. Findings will guide the adaptation of the existing mobile NAS tool for high-risk pregnant women. I will then test the usability, acceptability, and feasibility of the adapted mobile tool via surveys with 10 pregnant women receiving opioid agonist therapy (OAT) at Spokane Regional Health District’s Opioid Treatment Program and Evergreen Recovery Center. Finally, we will randomize 30 high-risk pregnant women seen at these facilities to either receive the adapted mobile NAS caregiving tool or usual care. We will compare these mothers on maternal drug relapse and OAT continuation, maternal-newborn bonding, length of newborn hospital stays, readmissions rates, breastfeeding initiation and duration, and postpartum depression and anxiety at 4, 8, and 12 weeks postpartum. Findings will serve as pilot data for a subsequent large R01 randomized controlled analog trial testing the efficacy of the adapted NAS caregiving tool in reducing poor outcomes for NAS-affected newborns and their mothers. My proposed research plan integrates activities, formal training, and mentorship from experts (Drs. Sterling McPherson, Hendree Jones, John Roll, Celestina Barbosa-Leiker, and Kim Johnson) in development, testing and implementation of substance use disorder treatment for perinatal women, mobile health interventions, and implementation of clinical trials in perinatal women with substance use disorders. This Mentored Research Scientist Development Award (K01) will build upon my previous training and allow me to pursue my long-term career goal of becoming an independent investigator with an established program of research focused on the development, implementation, and testing of interventions for substance using perinatal women and reduction of poor health outcomes for substance using perinatal women and their newborns.