Washington State University

NIH Research Projects · FY 2024 · 2020-06

PROJECT SUMMARY Since 2012 when the first human case of Middle East respiratory syndrome coronavirus (MERS-CoV) was confirmed, the World Health Organization has reported >2220 human infections and almost 800 deaths spread across 27 countries located in the Middle East, Europe, Asia, and the United States. Although dromedary camels are the known reservoir of the virus, there is limited knowledge on the mechanisms and factors associated with camel-to-human transmission, which remains the primary mechanism of human infections. Human outbreaks and the number of documented cases of MERS-CoV continue to grow in the Middle East and Asia; however there have, to date, been no documented cases of human disease in the eastern Africa countries where >65% of the world’s dromedary camels are found. This is despite evidence of prevalent MERS-CoV infection of camels in the region. The absence of human disease in East Africa may be explained by viral plasticity resulting in inefficient transmission and/or weakened virulence, or poor disease surveillance and reporting among the marginalized camel-owning nomadic pastoralist populations that inhabit remote arid lands of the regions. We will test these hypotheses by conducting integrated longitudinal cohort studies within a closed community of naïve pastoralists and their camel population that is known to sustain MERS-CoV circulation in Marsabit County, Kenya, in order to determine the maintenance and transmission of the virus among camels, zoonotic transmission to humans, and severity of human infections. To determine if the circulating MERS-CoV is genetically and phenotypically distinct from known virus clades in the Middle East and Asia, we will isolate the East African virus by collecting samples biweekly from an infant cohort (birth -1 year) of 211 camels, followed by culture and isolation of the virus to performed genotypic and phenotypic comparison with the known clade viruses To investigate whether a combination of weak surveillance and poor access to health care are responsible for absence of disease, we will follow-up for a year, a cohort of 573 camel handlers through biweekly visits, weekly telephone calls, and access to a toll-free number in order to intensively examine and test them for MERS-CoV disease. In addition, we will identify, test, and follow-up >4500 in- and out-patients with respiratory illness at Marsabit County Referral Hospital for 3 years. To assess the risk the virus poses to humans, we will determine the level of viral shedding in camels, and relate this to the incidence of zoonotic transmission, and types of camel contact that increase transmission risk. These studies will identify the type of virus circulating in East Africa, increasing knowledge about plasticity of MERS-CoV and its impact on zoonotic transmission and disease. By elucidating the frequency and mechanisms of zoonotic transmission, and progression to clinical human disease, we will define the risk the virus poses to this community at the frontline of a newly emergent virulent virus by virtue of their occupation and lifestyle, paving the way for development of improved surveillance and appropriate prevention and control strategies.

NIH Research Projects · FY 2024 · 2019-09

ABSTRACT: Alzheimer’s disease and related disorders (ADRD) is a growing public health concern for American Indian and Alaska Native (AI/ANs) people, but AI/ANs are profoundly underrepresented in ADRD research. For example, the National Alzheimer’s Coordinating Center database, an important resource for influential ADRD studies, includes just 162 AI/ANs out of 31,000 adults (0.5%). This is a troubling exclusion, since AI/ANs have high burdens of many ADRD risk factors, including hypertension, type 2 diabetes, traumatic brain injury, vascular brain injury (VBI), and stroke. The Cerebrovascular Disease and its Consequences in American Indians (CDCAI) study, conducted by our research group, is the only prospective cohort study with longitudinal data for rigorous evaluation of cognitive function, ADRD, and VBI in any US Native population. In its first wave of exams, CDCAI collected data on 818 elderly AI/ANs living on reservations in 3 primarily rural, geographic regions; the second wave of exams is underway. CDCAI is yielding seminal findings on ADRD risk factors and biomarkers in AI/ANs; 33% of the sample showed evidence of VBI, with men even higher. Yet the cohort is not representative of the national AI/AN population because 72% of AI/ANs now reside in urban areas. Urban A/ANs represent an “invisible tribe” that is largely absent from health research. One likely explanation is that urban environments feature distinctive risk and protective profiles, including lifestyle factors that exacerbate age-related cognitive decline, and protective factors such as better access to acute and specialty care and higher levels of education. Accordingly, we propose a novel study, URBan Native Elders (URBANE), to replicate CDCAI in a large cohort of urban AI/ANs with broad geographic representation. We will recruit 1,200 men and women ages 55 and older in 5 geographically diverse metropolitan areas with large AI/AN populations. Data collection will include MRI for all participants, and genetic testing for alleles associated with ADRD in the all-races population. We will expand CDCAI protocols to reflect state-of-the- art imaging and neuropsychological assessments. We will use protocols established by the CDCAI to estimate probable ADRD based on a single exam. The resulting analyses will establish the baseline prevalence of cognitive impairment, probable ADRD, and VBI and their associations with clinical, genetic, neuroimaging, behavioral, and lifestyle risk and protective factors. Of importance, this cross-sectional study will lay the foundation for future longitudinal research on ADRD. Our Specific Aims are to: Quantify the extent of probable ADRD and VBI as defined by brain MRI, and determine their associations with cognitive functioning in a diverse sample of urban AI/AN elders. 2) Evaluate associations between conventional risk factors and probable ADRD, and evaluate potential mediation of selected risk factors by VBI. 3) In an exploratory analysis, evaluate potential differences in our primary associations by sex; and evaluate education and access to healthcare as potential protective factors for cognitive impairment and probable ADRD in elders with VBI.

NIH Research Projects · FY 2024 · 2019-08

ABSTRACT American Indians (AIs) are disproportionately impacted by the current opioid epidemic. AIs experience an 88% higher prevalence of OUDs (1.5%) than the US general population (0.8%), and AIs are second only to non-Latinx whites in their experience of opioid overdose deaths (13.9 and 17.5 per 100,000, respectively). Fortunately, medication-assisted treatment (MAT; e.g., buprenorphine + naloxone) is highly effective for reducing opioid- related harm, including overdose, making it the gold-standard OUD treatment approach. A recent systematic review, however, showed a median retention of 56% at the NIDA-recommended 12-month treatment length. Further, most AIs with OUD do not attend traditional substance-use treatment (62%). This is concerning because treatment retention is strongly associated with mortality rate reduction. Although no studies have documented OUD treatment outcomes specific to AIs, research in Native communities has generally highlighted concerns about the cultural acceptability of the highly directive, Western medical substance-use treatment approaches (e.g., cognitive-behavioral therapy, 12-step programming) that are widely available. Further, previous research has indicated that AIs with substance use disorders want greater representation of Native staff and better integration of culturally adapted approaches in the services they receive. Culturally adapted approaches to treatment are associated with reductions in use and associated problems. However, there are currently no evidence-based, culturally adapted counseling approaches for AIs addressing MAT retention and opioid-related harm. Accordingly, we propose a 2-phase R61/R33 development and evaluation project in response to RFA-DA- 19-013 - “Responding to Opioid Use Disorders (OUD) in Tribal Communities in the Context of SAMHSA and CDC Funding.” This project will leverage recent federal OUD treatment initiative funding (SAMHSA TI-18-016, CDC-RFA-OT18-18030101supp) as a platform for culturally adapting substance-use counseling focused on improving MAT retention and reducing opioid-related harm within the Cherokee Nation Health System (CNHS). The 2-year R61 Phase will entail 2 parts. First, we will conduct a mixed methods inquiry to inform research methods and the community-specific, cultural adaptation of an existing, efficacious, harm-reduction counseling approach. Second, we will manualize and pilot the resulting Counseling for Harm Reduction and Retention in MAT at Cherokee Nation (CHaRRM-CN) together with a community advisory board comprising CNHS providers, staff, and patients as well as researchers from Cherokee Nation, Washington State University, and the University of Washington. The subsequent, 3-year R33 Phase will entail a 2-arm RCT (N=160) conducted within CNHS testing the efficacy of CHaRRM-CN in improving 6-month MAT retention, reducing substance-related harm and illicit opioid use, and increasing Native enculturation compared to a services-as-usual control condition (i.e., cognitive behavioral treatment).

NIH Research Projects · FY 2025 · 2019-01

Project Summary/Abstract (Parent Grant Application) Prostate cancer (PC) is the most common non-skin cancer in American men, with a lifetime incidence of 1 in 7, and also the second leading cause of cancer death in American men. Androgen receptor (AR) is the primary oncogenic driver of PC growth, survival and progression. AR-directed therapy is currently the principal treatment regimen. Despite initial response rates exceeding 90%, PC eventually relapses and progresses to fatal castration-resistant PC (CRPC), where reactivation of AR signaling occurs in a low-androgen environment. Recent introduction of FDA-approved next-generation antiandrogens, including enzalutamide (ENZ) and abiraterone acetate (ABI), have improved the CRPC treatment landscape, but emergence of drug resistance remains nearly universal, with no AR-targeted therapeutic options afterwards. These dismal facts underscore the pressing clinical need to identify new molecular targets and develop effective therapies to combat advanced PC. Through integrated analysis of publicly available clinical PC data sets coupled with functional studies in AR-positive PC cells, we propose monoamine oxidase A (MAOA), which synergizes with AR to promote PC, as an ideal therapeutic candidate to complement AR-targeted therapy in CRPC. We identified a novel reciprocal interaction between MAOA and AR in PC cells. MAOA expression is induced by androgen treatment; and conversely, MAOA silencing significantly reduces AR activity by lowering AR target gene expression and responsiveness to androgen stimulation in PC cells under both androgen-replete and depleted conditions as well as in a CRPC xenograft model. We showed significant positive co-expression of MAOA and AR target genes (PSA, TMPRSS2, NKX3.1) in multiple clinical data sets, including CRPC. Importantly, we found MAOA genomic amplification and/or epigenetic activation in 64% of samples in a CRPC data set, reinforced by elevated MAOA protein expression in our CRPC patient cohort. Additionally, we demonstrated that inhibition of MAOA by genetic or pharmacological approaches enhanced the growth-inhibiting effects of ENZ and ABI in androgen-sensitive, CR and antiandrogen-resistant PC cells. Based on these findings, we will test the hypothesis that MAOA synergizes with AR through reciprocal crosstalk and convergent downstream signaling to amply MAOA/AR effects promoting AR-driven PC growth and progression, and that co-targeting MAOA/AR is an actionable, effective strategy to treat CRPC and reverse antiandrogen drug resistance. To address this hypothesis, three aims are proposed. In Aim 1, we will elucidate the mechanistic basis of MAOA- AR reciprocal interaction in PC cells. In Aim 2, we will characterize the role of MAOA in regulating the development and progression of CRPC in xenograft models. In Aim 3, we will determine the efficacy of MAOA inhibitors for treating CRPC and reversing resistance to next-generation antiandrogens in vitro and in vivo. These studies will provide fundamental innovative insights into AR regulation in CRPC and illuminate a path toward the development of new combination therapy for advanced PC.

NIH Research Projects · FY 2026 · 2018-08

ABSTRACT Efficient repair of DNA base lesions, such as UV-induced cyclobutane pyrimidine dimers (CPDs) or DNA alkylation damage, is critical to maintain genome stability and prevent mutations that can lead to cancer. Repair of these abundant classes of DNA lesions is the responsibility of the nucleotide excision repair (NER) and base excision repair (BER) pathways. The importance of efficient excision repair is highlighted by the severe phenotypes of patients with Xeroderma pigmentosum (XP), which have inherited defects in NER genes, and the cancer predisposition of individuals with variants in key BER genes. While the basic mechanisms by which the NER and BER pathways repair DNA base lesions are well understood, how these excision repair pathways efficiently recognize and repair DNA lesions resident in eukaryotic chromatin is unclear. To address this question, we have developed a new high-throughput sequencing method known as MNase-CPD-seq, which provides an unprecedented snapshot of the dynamics of damaged nucleosomes in UV-irradiated cells. Our preliminary MNase-CPD-seq data indicate that damaged nucleosomes rapidly alter their rotational setting to move CPDs to more accessible Minor-Out rotational settings, and at later repair time points, alter translational positioning to expand linker regions between nucleosomes. These preliminary data form the basis of Aim I, where we will use MNase-CPD-seq to characterize the role of damage recognition factors, such as XPC or UV-DDB, and histone modifications in promoting repositioning of damaged nucleosomes in yeast and human cells and in vitro. In parallel, we also plan to develop related methods to map nucleosome dynamics associated with other classes of DNA lesions (i.e., UV-induced 6-4 photoproducts and N-methylpurine (NMP) lesions). In Aim II, we will test the hypothesis that ACR complexes in yeast and human cells are required for efficient repair of UV damage in chromatin by repositioning damaged nucleosomes during repair. We will also investigate how ACR mutants affect UV mutagenesis in UV-exposed yeast cells and human skin cancers. Finally, in the previous award we identified novel roles for histone PTMs in regulating distinct NER and BER pathways in chromatin. Aim III will build on the studies by characterizing the mechanisms by which histone acetylation regulates NER and BER in yeast chromatin, and the role of histone H3 K36 methylation by Set2 in yeast and SETD2 in human cells in regulating canonical and cryptic transcription coupled-nucleotide excision repair (TC-NER). Since ACR subunits (e.g., ARID2) and SETD2 are frequently mutated in human cancers, including melanoma, these studies have important implications to mechanisms of carcinogenesis and chemotherapeutic resistance in cancer.

NIH Research Projects · FY 2025 · 2018-07

1 Project Summary: Resistant bacterial infections impact animal health and welfare, but they also 2 have potential for causing negative human health consequences through transmission of resistant 3 bacteria or resistance genes through food or animal contact. The use of common classes of 4 antimicrobials in humans and animals increases the likelihood that drug resistance selected for in 5 animal species could impact humans and vice versa. Therefore, monitoring AMR in animals has 6 the potential to contribute to mitigation not only disease in animals but human infections as well. 7 Coordinated nationwide AMR monitoring in veterinary species for bacteria of importance in 8 major sectors of veterinary medicine is has been a recent phenomenon, and many data gaps exist. 9 Veterinary diagnostic laboratories are uniquely positioned to contribute to antimicrobial 10 resistance (AMR) monitoring through access to clinically relevant bacterial isolates and technical 11 expertise in laboratory testing. This project addresses these gaps by contributing to standardized 12 method development for whole genome sequencing to ensure the sensitivity and specificity is 13 appropriate for detecting the resistance of interest, improvement of the bioinformatics pipeline 14 with quality assurance and quality control criteria tailored to the veterinary diagnostic laboratory 15 and provide AMR data in agricultural and companion animal species. Each year of funding will 16 involve coordination with veterinary diagnostic source laboratories for acquisition of bacterial 17 isolates derived from clinically affected animals, including Salmonella, E. coli and 18 Staphylococcus pseudintermedius and other bacteria of interest, from across the US, performance 19 of whole genome sequencing of on a quarterly basis, and evaluation for the presence of 20 antimicrobial resistance genes. The data will be provided to FDA CVM Vet-LIRN for further 21 evaluation and future public availability. 22 23

NIH Research Projects · FY 2025 · 2017-09

Project Summary Although the integration of neurobiology into immunological research has just begun, it has already profoundly improved our understanding of immunology. By using the Caenorhabditis elegans model system, we have identified three specific neuro-immune regulatory circuits that are mediated by the G protein-coupled receptors OCTR-1, NPR-8, and NMUR-1, respectively. Our studies and others indicate that there could be a predetermined setpoint for internal immunity, like the setpoint for the human heart rate, and that the nervous system regulates immune responses to internal or external environmental changes and restores immune homeostasis by bringing immunity back to the setpoint. Considering this “setpoint for immune homeostasis” theory, we plan to investigate how the nervous system and immune system interact to achieve the resolution of infection and the specificity of innate immunity, two important but poorly understood issues in immunology. Previously, we have characterized the OCTR-1-dependent neuro-immune regulatory circuit in great detail and have revealed that NMUR-1 mediates the specificity of C. elegans innate immunity. Here, we propose studies to dissect the molecular and neuronal mechanisms underlying neural regulation of resolution in the context of the OCTR-1 circuit and to identify specific neurons, neural networks, and neural signals that regulate NMUR-1- mediated specificity of innate immunity. These studies will advance our understanding of how the nervous system controls both infection resolution and innate immune specificity as well as define key regulatory principles that govern neuro-immune relationships. Results from these studies could benefit the development of new treatments for infectious diseases and innate immune disorders.

NIH Research Projects · FY 2024 · 2017-06

In striated muscles, actin thin filament architecture is critical for efficient contractile activity, and alterations in thin filament integrity are linked to severe and often lethal skeletal and cardiac muscle diseases. Our long-term goal is to identify the components and molecular mechanisms regulating actin architecture in striated muscle during normal development and disease. Our short-term goal is to evaluate how actin-binding proteins of the tropomodulin family (e.g. leiomodin or Lmod and tropomoduin or Tmod) affect the formation and then the structure of the thin filament. We will test our recently proposed molecular mechanism for the Lmod/Tmod-dependent regulation of the pointed end of the thin filaments. We will also study the structural and functional consequences of Lmod binding to sides of the already formed thin filaments. Finally, we will establish mechanisms of regulation of Lmod functions. We propose three aims to identify underlying molecular mechanisms of the full spectrum of Lmod and Tmod functions: (1) to decipher the role of Lmod in the maintenance of proper thin filament lengths via pointed end regulation; (2) to establish the role of Lmod’s actin-binding sites in thin filament activation; (3) to test the hypothesis that Lmod functions are regulated by Ca2+. By employing high resolution cryogenic electron microscopy (cryo-EM) in conjunction with 3-dimensional nuclear magnetic resonance and Förster resonance energy transfer), we will recreate the full picture of Ca2+- dependent Lmod interactions with the thin filament and reveal the biological significance of these interactions. The robustness of structural models will be evaluated by monitoring of development and contractility of cardiac and skeletal muscles in knockout mice in vivo via the identification and utilization of mutations specifically affecting newly discovered Lmod’s and Tmod’s functionalities. To achieve these goals, we established a powerful multidisciplinary collaboration between the Kostyukova laboratory at the Washington State University (expert in protein structure, biochemical and biophysical properties of actin-binding proteins), the Gregorio laboratory at the University of Arizona (expert in the molecular, cellular and developmental biology of myofibril assembly) and the Galkin laboratory at the Eastern Virginia Medical School (expert in high resolution cryo-EM of actin complexes). Our data will provide a comprehensive identification of critical components of the regulatory mechanisms underlying thin filament assembly and maintenance in health and disease.

NIH Research Projects · FY 2024 · 2015-09

PROJECT SUMMARY OF THE OVERALL U54 CENTER Assessing the risk of adverse natural product (NP)-drug interactions (NPDIs) is of paramount importance due to the exponential NP sales growth in the US. This effort is challenged by relatively scant pharmacokinetic knowledge of individual NP constituents that precipitate these interactions and a lack of mechanistic understanding of NP effects on drug disposition. Investigations of NPDIs to address these knowledge gaps are complicated by the inherently large variability in phytoconstituent composition of supposedly the same NP, acquisition of sufficient high-quality NP study materials, and the diverse populations to which NPs are marketed, including the elderly. Our established Center of Excellence for Natural Product Drug Interaction Research (NaPDI Center) was created by the National Center for Complementary and Integrative Heath (NCCIH) in 2015 to address this public health problem. A cross-disciplinary research team composed of experts in clinical pharmacology, NP chemistry, biomedical informatics, and health communications led the charge to address the challenges unique to NPDI research. As detailed in this renewal application, we will advance the mission of the NaPDI Center by designing and executing new mechanistically-driven Interaction Projects, expanding the utility of our web-based data repository and public portal, and broadening dissemination of knowledge to national and international research communities. These activities will be accomplished through synergistic interactions between three established scientific Cores (Pharmacology, Analytical, and Informatics) and an overarching Administrative Core. The Pharmacology Core will expand an innovative method to select three to five high priority NPs to study as precipitants of potential clinically significant NPDIs. The Analytical Core will source and fully characterize the chemical composition of commercially available, representative NPs and provide NP study materials free of adulterants and contaminants for the Interaction Projects. The Pharmacology Core will design and execute rigorous in vitro studies to establish the mechanism of NP effects on drug disposition; predict the magnitude of NPDIs using physiologically-based pharmacokinetic modeling and simulation; and design and conduct clinical NPDI studies. The Analytical Core will provide quantitative analysis of object drugs, NP constituents, and emerging drug disposition biomarkers in the human pharmacokinetic samples. The interpreted data generated from the Interaction Projects will be transferred to the data repository developed and maintained by the Informatics Core. Major findings will be disseminated to scientific and other interested communities through our unique public access web portal perpetuated by the Informatics Core. All Cores will share the task of broadening Center outreach activities through targeted workshops and symposia at national and international scientific meetings. A Steering Committee composed of Core leaders and NCCIH officials will jointly oversee Center scientific progress with an External Advisory Panel of experts in relevant fields selected by NCCIH.

NIH Research Projects · FY 2025 · 2014-09

PROJECT SUMMARY/ABSTRACT The world's population is aging. The resulting prevalence and ability to provide quality care for older individuals with Alzheimer's disease and related dementias (ADRDs) and other chronic illnesses is a challenge our society must address. Our vision is to address this challenge by providing a diverse body of undergraduate students with the scientific, clinical, and research experience needed to understand health-assistive technology and design technological solutions that aid with the challenges of aging and improve human health. Undergraduates in neuroscience, psychology, sociology, computer science, and engineering (MSTEM) programs as well as those in healthcare-related disciplines need a strong multi-disciplinary background to be truly prepared for research in applying technology to gerontology issues. The objective of this renewal application is to continue to enhance and lead a research training program for undergraduate students in the fields of gerontology and technology-based assistive environments. This will be done through course work, summer research programs, online materials and professional symposia to help other institutions develop similar programs. The ultimate goal is to bring up a diverse generation of new graduate student researchers and innovators who understand the need of continued work in the field for addressing the aging population issues and begin their research careers prepared for gerontechnology oriented research. To accomplish our goal, we will refine and offer a gerontechnology class that is geared toward multidisciplinary undergraduate students (Aim 1). We will also refine and offer a gerontechnology-focused summer undergraduate research experience (GSUR) program that will provide a team-based research opportunity for highly-qualified students (Aim 2). To broaden the impact of the training program, we will offer mentoring support for senior capstone projects and independent and clinical training projects related to gerontechnology (Aim 3). Finally, we will broaden the impact of our program by disseminating training materials through online classes, Youtube videos, and podcasts, and presenting methods and results of the training program at high-visibility gerontology and technology meetings (Aim 4). In all of these efforts we will recruit and involve a diverse student body, including women in STEM, minorities, persons with disabilities and individuals from disadvantaged backgrounds. The proposed program is innovative because Gerontechnology-related undergraduate programs with a true multi- disciplinary core are rare. The combination of serving both the local student body, summer students from other programs, and individuals from outside the university through online materials, open seminars, and workshops will bolster the quality, quantity, and diversity of highly prepared upcoming graduate researchers. The project is significant because it will introduce many undergraduates to the issues faced by our society in the coming decades, as well as prepare many of them to help develop new approaches to health care through melding technology with traditional medical approaches.

NIH Research Projects · FY 2024 · 2014-09

Abstract In our parent R 01 application, we have shown that the presence of dopants in HA influences the bone remodeling process and phase stability in coating while improving coating interfacial mechanical properties processed via laser and plasma. Specific dopants like magnesium (Mg), zinc (Zn), and silicon (Si) in their oxide form help in osteogenesis and angiogenesis. However, those dopants have limited influence on osteoinductivity or osteoclastogenesis, and both are needed to minimize aseptic loosening. The objective of this follow-up application is to understand osteogenesis mechanisms via in vitro gene expressions and in vivo studies using rat and rabbit distal femur models of doped HA-coated implants with plant-derived compounds (PDCs) such as gingerol from ginger, allicin from garlic, and carvacrol from oregano. It is envisioned that PDCs will be responsible for osteoinduction and reduced osteoclastogenesis to improve bone tissue-material interactions, minimize aseptic loosening and implant migration. Over 1 million total joint arthroplasties have been performed in 2019 in the United States alone. Aseptic loosening is still considered one of the most common failure modes in these surgeries' coming days. Metal implants such as Ti6Al4V are widely used for arthroplasties due to their excellent biocompatibility in vivo. Although Ti6Al4V is osteoconductive, cementless implants for joint replacement depend on stringent initial mechanical stability for bone ingrowth/apposition to occur. Initial implant integration and avoidance of late aseptic loosening are even more challenging in the revision scenario, in which the bone-bed is often sclerotic and dysvascular. We hypothesize that: 1) addition of PDCs with selected dopants in HA coating will modulate release kinetics and control bone remodeling in vivo; 2) PDCs will locally increase bone density and introduce osteoinductivity, especially for patients with compromised bone in revision surgeries; and 3) an easily introduced oxide layer between the metal substrate and HA coating will maintain stronger bonding between the metal implant and the ceramic coating. Our design goals are: 1) improve bioactivity and introduce osteoinductivity to minimize healing time and 2) enhance interfacial mechanical properties between the HA coating and the implant to increase coating in vivo lifetime. We propose the following two specific aims: (1) Aim 1: Understanding of gene expressions for PDC added doped HA coatings on Ti6Al4V; (2) Aim 2: Evaluate effects of PDC release from doped HA-coated Ti6Al4V on early-stage bone cell – materials interactions and in vivo biological response. The following outcome measures will judge this project's success: quantitative and qualitative control of PDC added doped HA coatings on Ti6Al4V in terms of enhanced osteoinductivity, uniformity in microstructure leading to higher interfacial mechanical strength, improved cellular differentiation, and bone-tissue integration. This program's scientific understanding will improve the long-term fixation of cementless implants and minimize aseptic loosening.

NIH Research Projects · FY 2025 · 2012-09

1 Project Summary: The goal of the CVM Vet-LRN Veterinary Diagnostic Laboratory Program is 2 to complement, develop, and use university, state and federal veterinary diagnostic laboratory 3 resources and expertise in order to increase national laboratory capacity to detect, respond to 4 and recover from a catastrophic animal food/feed contamination event, either microbial or 5 chemical. The program's funding allows provision for equipment, supplies, and personnel; 6 training in standardized testing methodologies; participation in proficiency testing in those 7 methodologies; participation in method enhancement activities to extend analysis capability; and 8 analysis of surveillance and emergency outbreak samples. Vet-LIRN laboratories nationwide 9 strengthen the overall food safety system through increased capacity and capability to detect 10 adulterations affecting animals raised for human consumption or companion animals consuming 11 ingredients used in both animal and human food products. The Washington Animal Disease 12 Diagnostic Laboratory (WADDL) at Washington State University is well positioned to effectively 13 function as a regional laboratory in the CVM Vet-LIRN Veterinary Diagnostic Laboratory 14 Program. Since 1974 WADDL has provided full laboratory diagnostic services (approximately 15 300,000 tests annually), including pathology, microbiology and toxicology, to veterinarians and 16 animal owners throughout the Pacific Northwest. WADDL actively participates in multiple 17 national laboratory networks (Vet-LIRN, USDA-NAHLN, CDC-LRN) and is well practiced in 18 standardized methods, equipment, proficiency testing, electronic reporting, and national disease 19 surveillance programs emphasizing early detection and response. WADDL has a mature 20 laboratory quality system based upon international ISO 17025 laboratory standards, being fully 21 accredited by the American Association of Veterinary Laboratory Diagnosticians laboratory 22 accreditation program since 1978. Using FDA-Vet-LIRN funding during 2012-2017 WADDL 23 successfully participated in sample analysis for surveillance and consumer report investigations, 24 provided proficiency testing, electronic reporting, and quality system implementation, and 25 participated in special projects on companion animal salmonellosis, and methods development 26 and matrix extension for analytical chemistry testing. The laboratory is fully capable and highly 27 competent to participate in FDA/Vet-LIRN key project areas of sample analysis, providing 28 analytical data to support regulatory use, and participating in additional projects regarding 29 method development, validation and matrix extension.

NIH Research Projects · FY 2025 · 2010-09

Maternal obesity, AMPK and Developmental Programming Over 30% of pregnant American women are obese and an additional 30% are over-weight, conditions which negatively affect fetal development with long-term consequences for offspring health, including pre-disposition to obesity and type 2 diabetes (T2D). The underlying mechanisms remain poorly defined. Skeletal muscle (SM) accounts for >30% body weight and is a key tissue for the oxidation of fatty acids and glucose, as is brown adipose tissue (BAT). Under the support of our previous award, we demonstrated that maternal obesity (MO) elicits early onset of fibrotic and fatty infiltration (FFI) in offspring SM and BAT, which impairs their functions and programs metabolic disorders in offspring. We found that AMP-activated protein kinase (AMPK), a known target for metformin, is robustly inhibited due to MO, correlates with FFI and worsened offspring SM/BAT functions. In the early embryo, both myogenic and fibrogenic cells are derived from progenitor cells (PCs) in the dermomyotome. While the embryonic myogenic cells are the sources of both myogenic and brown adipogenic cells, embryonic fibrogenic cells are sources of fibrogenic and white adipogenic cells in offspring SM/BAT. Because of this, our previous studies point to embryonic origins for developmental abnormalities of offspring SM/BAT due to MO, but this is yet to be examined. Using single cell RNA sequencing (scRNA-seq), we found that embryonic myogenesis is attenuated in E9.5 MO embryos. We hypothesize that MO suppresses AMPK, which inhibits myogenic commitment and drives uncommitted PCs to fibrogenesis during embryonic development, programming FFI in offspring SM/BAT. Accordingly, we have three specific aims: 1) Study AMPK inhibition due to MO in impairing embryonic myogenic commitment; 2) Evaluate AMPK in linking MO to enhanced embryonic fibrogenesis; and 3) Analyze AMPK as a target for improving embryonic SM/BAT development of MO and the resulting offspring SM/BAT functions. We will use single cell “omics” for analyzing embryonic tissues, and embryoid body culture for mechanistic exploration. Knowledge obtained will identify molecular targets for therapeutics to improve embryonic SM/BAT development and subsequent metabolic health of MO offspring, helping the increasing populations of obese mothers to deliver healthy children.

NIH Research Projects · FY 2025 · 1989-09

Project Summary The landmark Institute of Medicine report, “Emerging Infections: Microbial Threats to Health in the United States" highlighted the importance of the epidemiology, pathogenesis, and improved control for zoonotic infectious diseases in natural animal hosts—60% of all human pathogens are directly transmitted from or emergent from animal reservoirs. Understanding the behavior of zoonotic pathogens, including mechanisms of persistence, evolution of virulence, and genetic change underlying transmission phenotypes, is now widely recognized as critically important to addressing emerging infections. In addition, the importance of zoonotic pathogens and normal microbial flora of animals in emergence, formation of reservoirs, and spread of antimicrobial resistance has been highlighted in recent reports by both CDC and WHO. This Post-doctoral Training Program specifically prepares a scientific workforce prepared to and capable of addressing critical knowledge gaps in infectious diseases and antimicrobial resistance. The Training Program focuses on the integrated training of two types of post-doctoral fellows: (i) clinically-trained veterinarians with either residency training or a MPH (3-5 years post-DVM); and (ii) recent PhD graduates with research experience in cell biology, genetics, epidemiology, immunology, or microbiology (0-2 years post-PhD). Training for those who enter the program with DVM degrees is guided by two principles: (i) a strong basic sciences foundation is indispensable and is attained with targeted coursework; and ii) a minimum of 3 years of dedicated research is required to build the basis for career progression to independence. Training for post-doctoral fellows with PhD degrees is tailored to their backgrounds but emphasizes research directed at critical knowledge gaps in zoonotic infectious diseases and/or antimicrobial resistance. In addition to education on the responsible conduct of research, all trainees receive instruction on experimental design and data interpretation that generates rigorous and reproducible results. As the Training Program recruits primarily entry-level post-doctoral fellows, 3 years of fellowship commitment is required. This integration provides a dynamic training environment where post-DVM fellows develop the basic research skills required to address and solve complex diseases while post-PhD fellows develop specific expertise with microbial pathogens and a broader and deeper understanding of the global health relevance and impact. Program data strongly supports that the Training Program is addressing a significant gap with strong demand and preparing both types of entry level fellows for successful careers.

NIH Research Projects · FY 2026 · 1977-08

Project Summary/Abstract: The cycle of the seminiferous epithelium is a characteristic of spermatogenesis in mammals. The cycle serves to distribute the developmental stages of spermatogenesis in constant periodic intervals along the seminiferous tubules. The function of the cycle is to assure a constant source of spermatozoa over the relatively extended developmental time period. We have previously shown in the mouse testis that the cycle is initiated by pulses of retinoic acid (RA) originally generated by the Sertoli cells at stages VIII and IX of the cycle. The pulse of RA forces the undifferentiated spermatogonia to irreversibly enter the differentiation pathway. At stages VIII and IX we have shown in the adult mouse that the Sertoli cells demonstrate an increased abundance of hundreds of transcripts that allow for multiple functions such as RA synthesis, junction formation, uptake of residual bodies, spermiation, and others. In the absence of germ cells there are no cyclic activities of Sertoli cells detectable along the tubules, thus, accentuating the requirement for interactions between Sertoli and germ cells for normal spermatogenesis. We propose to examine the influence of spermatogonia at stages VIII and IX on the transcriptome of Sertoli cells and their functions during the development of complete spermatogenesis. We propose that this influence on the transcriptome and on the generation of the RA pulse is partially or wholly a result of Sertoli-germ cell interactions through the Notch signaling pathway. We will test this hypothesis by examining the influence of germ cells with Notch ligand gene deletions on the transcriptome and RA biosynthesis in normally developing Sertoli cells and in Sertoli cells with Notch receptor deletions. We will utilize a new technology designated capped small RNAseq or nTIseq to obtain information on active promoters and transcriptional start sites in the presence and absence of an intact Notch signaling pathway. We will pay special attention to transcripts regulating RA biosynthesis such as RDH10. The results of this study will clarify the interactive roles of germ and somatic cells in spermatogenesis and possibly implicate the Notch pathway in the etiology of male infertility.

Other NSERC · FY 2024

Biofuel, Bioproducts, Membrane bioreactor, Anaerobic digestion, Pilot system, Renewable energy, Resource recovery, pretreatment, Sludge treatment, Lignocellulosic biomass