Purdue University

NIH Research Projects · FY 2025 · 2022-09

Alzheimer's Disease (AD), AD-related dementias (ADRD) and other neurodegenerative diseases such as Parkinson’s disease (PD) exhibit pathogenic gene-environment interactions (GxE) with synergistic effects of exposure to an environmental chemical/pathogen and genotype. Recent progress in next- generation sequencing has expedited the discovery of genetic risk factors associated with Alzheimer's disease, AD-related dementias and Parkinson’s disease. Yet, identified genetic factors only account for a fraction of patients, and not all patients with the identified genetic risks develop disease. This is exasperated for AD/ADRD/PD as it is convoluted with many covariates over a person’s lifetime. To elucidate the contribution of GxE to disease we propose an approach based on the principles of latent-persistent effects of environmental neurotoxicants; and the developmental origins of adult disease hypothesis. We seek to test the hypothesis that persistent neurotoxicity is due to exposures altering self-perpetuating homeostatic processes that give the resiliency and perpetuity to the adverse toxicological processes by either genetic and/or epigenetic means. Specifically, we will collect samples from same-sex siblings in northern Indiana that phenotypically differ in AD/ADRD/PD relevant cognitive and motor dysfunction phenotypes; reprogram blood cells into human induced pluripotent stem cells (hiPSCs); differentiate them into cortical and midbrain lineages; and characterize their vulnerability to induction of a persistent neurotoxic state caused by neurodevelopmental exposures to environmental hazard/toxicants relevant to this region and history. We expect to identify a persistent neurotoxic state with a conserved response shared across individuals at the level of the genetic/epigenetic pathways evoked. 'Age-related signaling' networks have been defined as central pathways regulating healthy lifespan and aging and are thus expected as a shared network feature conferred by GxE risks. Our innovative approach will leverage single-cell genomics approaches and pathway analysis to identify mechanisms activated by subject-specific GxE to cause a persistent neurotoxic state with only a small subject size and low FDR. The following aims will test our hypothesis and validate the shared signaling networks: Aim 1: Identify subject by toxicant interactions contributing to persistent neurotoxicity using human cells derived from paired Alzheimer's disease, AD-related dementias and Parkinson’s disease and healthy cohort with comparable exposure histories; Aim 2: Identify genetic pathways associated with establishing a persistent neurotoxic state via single-cell genomics and bioinformatic comparisons to population level data; and Aim 3: Validate the genetic pathways of GxE induction of persistent neurotoxicity with in vivo and in vitro models. This work seeks to understand how past environmental exposures influence AD/ADRD/PD disease risk and incidence by utilizing an interdisciplinary academic- community partnership to study elderly subjects at risk for AD/ADRD/PD. The findings may also identify targets of the GxE interactions that contribute to the increased risk of Alzheimer's disease, AD-related dementias and Parkinson’s disease over our lifetime.

NIH Research Projects · FY 2025 · 2022-09

Project summary Interactions between human mesenchymal stem cells (hMSCs) and their environment are a main factor in the function of these cells. Although the importance of cell-material interactions is well established, it has been diffi- cult to characterize this complex interplay, especially in vivo. During in vivo experiments, a stem cell treatment's efficacy can be assessed, but the underlying cellular processes and change in the surrounding microenvironment that leads to these outcomes remain a black box. Positive outcomes may result from these experiments even if there is loss of function or integrity in the tissue due to failed cell-material interactions, making aspects of the stem cell treatment ineffective and potentially unnecessary. Since real-time measurement of these interactions has not been realized in vivo, in vitro models provide an alternative approach to measuring cell-material inter- actions in both 2D and 3D culture. The use of scaffolds to mimic aspects of native tissue provide controlled environments where cell-material interactions can be quantitatively characterized. These scaffolds are used for 3D cell encapsulation and are designed to be remodeled by cells. This creates a feedback loop where the cell remodels the pericellular region and responds to the dynamically changing cues in the environment. Real-time characterization of these dynamic cell-material interactions continues to be a challenge. We propose to char- acterize dynamic cell-material interactions by measuring real-time hMSC-mediated scaffold remodeling using microrheological characterization and the resulting cellular processes using cell staining and inhibition. We will use an hMSC-laden synthetic hydrogel scaffold that mimics aspects of native microenvironments to present cues to cells. To characterize hMSC function, we will use techniques including cell staining and pharmacological inhibi- tion of molecules for cellular contractility and matrix adhesion. Our unique approach will characterize the scaffold microenvironment in real-time during cell-material interactions. We will use multiple particle tracking microrhe- ology (MPT) to measure hMSC-mediated scaffold remodeling and degradation. This technique quantifies the spatio-temporal evolution of the rheology in the pericellular region, which is part of the feedback loop that defines cell-material interactions. Together, these measurements will provide a relationship between cellular function and cell-engineered pericellular rheology as the complexity of the scaffold microenvironment is increased. The pro- posed research program will focus on characterizing cell-material interactions during specific critical processes that are not fully understood. The processes we will study are (1) cellular adhesion, (2) hMSC motility in re- sponse to scaffold viscoelasticity and (3) hMSC-material interactions when signaling molecules are presented in the environment. The proposed work will support the overarching goal of understanding the fundamentals of cell-material interactions and the influence on basic cellular processes.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY The capability to precisely control behaviors of biomolecules in living cells is a challenging task. Current methods can be classified into chemical-based and laser-based approaches. For example, small molecule inhibitors or activators can be introduced into the biological system for manipulating enzyme activities. However, it is impossible to control the interaction locations with high precision, which poses off-target effects. Protein control using genetic methods such as gene silencing or editing might selectively impact a targeted protein, but requires transfection and incubation, and cannot be performed in real-time. Optical techniques such as optical tweezers can manipulate small targets at the laser focus, but can only interact with a few targets at a time. Current laser manipulation and ablation methods usually require a pre-acquired image together with a manual selection of target locations on samples. This method is not only time-consuming but also unsuitable to apply to highly dynamic living biological samples. Optogenetic methods can control neuron functions using light radiation and light-sensitive ion channels, but only at the single-cell level. There’s no existing technology that can select molecular targets in cells and control only these targets at sub-micron resolution in real-time. In this application, we develop a real-time precision opto-control (RPOC) platform that can selectively and precisely control biomolecules only at the desired interaction site using lasers. RPOC is based on a high-speed laser scanning system. First, during the laser scanning, an optical signal is generated at a specific pixel from the target molecule. Then, this optical signal will be compared with a preset threshold and to send out an electronic signal to control an acousto-optic modulator which is used as a fast switch to couple another laser beam to interact with the same pixel. The optical signal detection, processing, and laser control happen within 20 ns, much faster than the pixel dwell time. Digital logic circuits will also be designed with the comparator circuits to control the interaction laser beam based on the logic output from multiple signal channels. We will use photo- switchable proteins and design photo-convertible inhibitors and activators to demonstrate precision control of enzyme activities on site. Furthermore, we will use multiple continuous-wave lasers and acousto-optic tunable filters to design a portable and multicolor RPOC that can operate outside an optical lab. RPOC can accurately control and manipulate biomolecules in real-time without affecting other biomolecules in the system. It is highly chemical selective since the optical signal can be selected from fluorescence, Raman, or absorption signals. It will allow biologists to control and interrogate only the biomolecules of interest during laser scanning without affecting other parts of the sample with sub-micron precision. RPOC would be widely applied to study enzyme activities in cells, understand organelle interactions, improve controlled-release of drugs, and perform precision neuromodulation.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Malaria is one of the most serious public health problems in sub-Saharan Africa. School-age children are most commonly infected with malaria parasites with an estimated 200 million at risk. Malaria screening for school- age children in endemic countries is critical in two aspects: malaria transmission and educational performance (human capital investment). Malaria rapid diagnostic test (RDT)-based interventions have shown to be effective, but mass screening with malaria RDTs on a routine basis is expensive and impractical. As a result, school-age children are often excluded. In this respect, risk stratification (prescreening) for malaria RDTs can play a critical role in the diagnosis and management of malaria. We hypothesize that a combination of blood hemoglobin level and acute undifferentiated febrile illness assessments can risk-stratify school-age children who will benefit from malaria RDTs and avoid unnecessary RDTs. Malaria infections in school-age children are strongly associated with anemia. Thus, noninvasive blood hemoglobin level readings can be highly beneficial for identifying asymptomatic (undetected) afebrile malaria infections. We will take advantage of our recently developed mHealth method that can reliably predict blood hemoglobin levels from digital photographs of the inner eyelid taken by a low-end smartphone. In Aim 1 (R21 phase), we will perfect an mHealth blood hemoglobin computation algorithm applied to school-age children (6 to 15 years of age) in Rwanda. The proposed machine learning approach will hybridize deep learning and statistical learning to accurately and precisely measure blood hemoglobin content among school-age children using an unmodified smartphone. In Aim 2 (R33 phase), we will develop an mHealth risk-stratification model to determine the need of malaria RDTs among school-age children. We will investigate the added value of mHealth blood hemoglobin assessments in identifying patients who will benefit from malaria RDTs and will need confirmatory malaria diagnosis. We will further formulate an advanced risk-stratification model that can forecast molecular test-confirmed malaria. In Aim 3 (R33 phase), we will implement an mHealth application integrating malaria risk stratification with the existing electronic health record (EHR) system. We will incorporate the mHealth technology into an Android- based EHR-integrated mobile application for community health workers (CHWs) and health facilities in our study settings. We will also include a digital reporting platform to replace paper-based patient data collection for CHWs and allow for automatic transmission into the currently used EHR system in our study settings. After successful completion, we expect to improve malaria diagnosis and management among school-age children, by empowering CHWs and health facilities with less hardware-dependent mHealth technologies. The proposed data-driven and connected mHealth technologies can maximize the nationwide scale-up of cost-effective malaria diagnosis and management in Rwanda, potentially offering mobility, simplicity, and affordability for rapid and scalable adaptation in other resources-limited settings.

NIH Research Projects · FY 2025 · 2022-09

Project Summary / Abstract Invasive urinary bladder cancer (invasive urothelial carcinoma, InvUC) is lethal in 50% of patients. Immune checkpoint inhibitors (ICIs) can cause dramatic remission of advanced InvUC, but only ~20% of patients have this level of benefit. Pre-clinical animal models are critical for research to improve ICI outcomes, but experimental models lack many of the hallmark features of human cancer and are poor predictors of outcomes in humans. To address the gap in relevant animal cancer models for immunotherapy research, we will study dogs with naturally-occurring InvUC as canine InvUC closely mimics the human condition in pathology, molecular features including luminal and basal subtypes, clinical presentation, local invasion, and frequent metastasis. The proposed work will strengthen the canine InvUC model by defining immune cell responsiveness and ICI outcomes, with comparison to human studies. Our long range goal is to improve the outlook for people with InvUC. The objective of this proposal is to address the gap by determining the suitability of canine InvUC to serve as a model to improve ICI therapy in humans. The central hypothesis is marked similarities will exist between dogs and humans in the immune cell responses in InvUC, and ICI therapy effects including immune adverse events, antitumor activity, immunological responses, and predictors of treatment success and failure. Some differences between dogs and humans are expected, with these also being informative. The hypothesis is formulated and based on strong evidence in the literature and preliminary data. The rationale is that demonstrating the shared immune cell responses and ICI effects between dogs and humans with InvUC will allow the canine model to be fully employed to improve ICI therapy for humans. The objective will be accomplished through two specific aims: (1) determine similarities and differences between dogs and humans in the immune cell responsiveness to InvUC, and (2) determine the safety and antitumor activity of a canine PD-L1 antibody and predictors of success and failure in dogs with InvUC with comparison to findings in humans. The approach will be to: (1) perform dog-human comparison of InvUC through analyses of RNA-seq, scRNA-seq, WGS, and CITE-seq data, and (2) conduct a clinical trial of an ICI, our canine PD-L1 antibody, in dogs with InvUC to assess antitumor activity, pharmacokinetics, adverse event profile, and correlative sequencing and clinical data to predict outcomes, and to compare results to those from human PD- L1 antibody trials. The expected results will define shared immune cell responses and ICI effects in dogs and humans, expand the understanding of predictors of ICI therapy, and justify use of the canine model to improve ICI and other immunotherapies in humans. Samples from dogs in the ICI trial will also be made available for other immune, microRNA, and microbiome research by our collaborators and beyond. We look forward to contributing to the Canine Cancer Immunotherapy Network and depositing our study data in the NCI's Integrated Canine Data Commons, to complement two InvUC data sets that we have previously deposited.

NIH Research Projects · FY 2025 · 2022-08

Poor diet is a leading cause of preventable death and diseases, as well as preventable healthcare costs in the United States. Despite the importance of following a healthy dietary pattern, most U.S. adults do not meet national dietary guidelines and are either overweight or obese. There is a critical need for "just-in-time" (JIT) interventions to improve diet and eating behaviors as they occur. To maximize impact, JIT interventions should only be delivered when an individual is receptive, particularly when dietary quality is poor. However, which aspects of the food environment and dietary behavior have influence on dietary intake and quality are unknown, and how they relate to JIT intervention receptivity is unexplored. This would require collecting and analyzing near-continuous data about one's diet in the context of daily life, where behavior actually occurs, which is very challenging for researchers and burdensome for participants. Advances in wearable sensor technologies, equipped with novel computational methods could provide a pathway to capture and analyze the various exposures and patterns in the eating environment to fill this gap. The overall objective of this proposal is to create an integrated system of wearable sensor and computational methods to discover food environment exposures related to dietary quality that influence JIT intervention receptivity. Motivated by this vision, the objectives of this research include: 1) develop novel edge computing hardware and software for privacy-preserving compressive image capture and transmission, 2) develop new collaborative compression and analytics together with unsupervised continual learning to understand eating behavior, 3) determine whether sensed aspects of the environmental context during eating relate to dietary quality and receptivity to JIT interventions, particularly when the dietary quality is poor. The project is a collaborative effort combining expertise in wearable electronics, image processing, dietary patterns, and behavioral science.

NIH Research Projects · FY 2024 · 2022-08

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NIH Research Projects · FY 2025 · 2022-08

Project Abstract: Gonorrhea is a sexually transmitted disease caused by the bacterial pathogen Neisseria gonorrhoeae that colonizes urogenital, anal, and nasopharyngeal tissues. Locally in the United States the Centers for Disease Control and Prevention (CDC) reported a 67% increase of gonorrhea cases between 2013 – 2017 with >550,000 cases in 2017 alone. N. gonorrhoeae wreaks havoc on world health care systems causing pelvic inflammatory disease, infertility and ectopic pregnancies. The bacteria can also be transmitted from mother to child during birth and lead to blindness. If left untreated N. gonorrhoeae can cause gonococcemia resulting in skin infection, arthritis or endocarditis. Pathogenic gonorrhea strains are increasingly resistant to common front-line antibiotics. The WHO surveillance program reports resistance to most available antibiotics. Rampant resistance has caused the CDC and the World Health Organization each to classify N. gonorrhoeae as a superbug and a future with an untreatable gonococcal infection is a real possibility. Thus, there is significant unmet need to identify novel targets and molecules with therapeutic potential. Studies proposed in this application build upon discoveries that FDA- approved carbonic anhydrase inhibitors (CAIs), such as acetazolamide and ethoxzolamide, display potent antimicrobial activity, in an applicable clinical range, against N. gonorrhoeae. CAIs, and analogs we have designed, also have no antimicrobial effect on commensal bacteria reducing the potential for problematic dysbiosis caused by antibiotic treatment. We have shown that the molecules exhibit their antibiotic effect by inhibiting the carbonic anhydrase from N. gonorrhoeae and have validated N. gonorrhoeae carbonic anhydrase (NgCA) as a viable anti-gonococcal therapeutic target. Our team has improved the potency of the CAI-based inhibitors from 4 µg/mL to 0.5 µg/mL. This proposal will continue lead optimization of CAI-based analogs using structure-based design while incorporating modifications to improve permeability into the Gram-negative cell. Molecules will be assessed in in vitro antimicrobial assays and prioritized analogs will progress to in vitro pharmacokinetic (PK) and pharmacologic profiling. Finally, top performing analogs will be assessed for in vivo efficacy in various gonorrhea mouse models as well evaluated the in safety and pharmacokinetic assay to support future lead selection and investigational new drug enabling studies.

NIH Research Projects · FY 2026 · 2022-08

Project Summary We propose to develop a community engaged research program in Martinsville, IN, a community of 11,000 people that overlays four groundwater contamination sites, including a U.S. Environmental Protection Agency- designated Superfund site. The total size of the groundwater contamination is over 60 acres and lies within a single aquifer. The contaminants are chlorinated volatile organic compounds (CVOCs), primarily tetrachloroethylene (PCE) and trichloroethylene (TCE), thought to originate from several dry cleaning and metal degreasing operations. An activated carbon filtration system has been in operation since 2005 to remove PCE/COVCs from the municipal water, which now meets EPA drinking water standards. However, a recent report from the Agency for Toxic Substances and Disease Registry concludes that people’s health may be harmed by breathing indoor air contaminated via vapor intrusion, the migration of volatile compounds from contaminated groundwater and soil into buildings above. The long-term goal of our research is to understand health effects of PCE exposure in communities, and to empower community members’ participation in environmental health decision-making. The project will be led by a transdisciplinary academic team in a strong partnership with two community based organizations. This team will partner with an existing, engaged Community Advisory Board (CAB), to accomplish three aims. Aim 1 will assess the community’s concerns and perceptions about the contamination and associated health risks, and how that changes across the 5-year project. Aim 2 will quantify current exposure to PCE/CVOCs in exhaled breath and indoor air from 300 residents using a cutting edge assessment tool, proton transfer reaction time-of-flight mass spectrometer (PTR-TOF-MS). Through modeling we will estimate cumulative exposures in residents and investigate associations of CVOC mixtures with visual and cognitive functions, which are hypothesized to be a sensitive non-cancer endpoint for PCE exposure. We will also address the community’s concerns about elevated cancer rates using Indiana state cancer registry data. In Aim 3, we will design and implement a targeted educational campaign to promote PCE/CVOC testing and installation of mitigation systems when residential contamination levels exceed exposure limits, with a focus on promoting property owners’ and landlords’ participation in vapor intrusion testing and remediation activities. With the CAB, we will co-design the action plan, identifying and responding to emerging barriers, through a Developmental Evaluation (DE) process utilizing a Theory of Change (TOC) model informed by the evaluation team’s qualitative and quantitative analysis of data on stakeholder concerns and responses. Throughout the process of community assessment and education, we will evaluate the effectiveness of the process, identifying pathways for moving from research to effective actions.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Human induced pluripotent stem cells (hiPSCs) can be differentiated to cells in all three germ layers (ectoderm, mesoderm, and endoderm), providing an invaluable cell source for basic research and translational applications. While recent years have witnessed breakthroughs in lineage-specific differentiation of hiPSC, the effect of matrix stiffness, viscoelasticity, and integrin ligand presentation during multi-stage exocrine pancreatic organoid (exoPO) development remain largely unexplored. Furthermore, current three-dimensional (3D) matrices for hiPSC culture and differentiation do not provide sufficient controls over matrix biophysical properties (e.g., viscoelasticity, stiffness) and biochemical motifs (e.g., cell-adhesive ligands). Additionally, no prior work has employed dynamic xeno-free hydrogels to study the effect of matrix mechanics and cell- adhesive ligand presentation on the development of hiPSC-derived exoPO. We hypothesize that exoPO differentiation can be drastically improved by presenting the cells with fine-tuned 3D matrix properties during the developmental stages. To achieve this goal, we will develop a viscoelastic dynamic double network (DDN) hydrogel platform with unprecedented tunability in matrix mechanical properties and biochemical motifs. Specifically, we will control matrix stiffness by forming an elastic hydrogel network with inverse Electron Demand Diels-Alder (iEDDA) click reaction. We will tune matrix stress-relaxation through a set of linear polymers complexed by reversible boronate-diol bonding. Uniquely, the elastic iEDDA click hydrogel network will be engineered to exhibit tunable hydrolytic degradation. On the other hand, the viscoelastic network will allow conjugation of cell adhesive ligands to permit viscoelasticity mediated engagement of integrins. With this viscoelastic DDN hydrogel platform, we will define the impact of matrix viscoelasticity, stiffness, and integrin ligand presentation on multipotent pancreatic progenitor cell differentiation and exoPO formation. In Aim 1, we will study the role of matrix viscoelasticity on pancreatic progenitor differentiation. In Aim 2, we will describe the requirements of matrix stiffness during pancreatic progenitor differentiation. In Aim 3, we will identify the role of cell adhesive ligands on pancreatic ductal/acinar cell specification. In the long-term, this project will produce a dynamic hydrogel platform to advance the use of chemically-defined matrices as xeno-free artificial stem cell niches for organoid development and tissue regeneration applications.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Ubiquitin (Ub) is a principal post-translational modifier crucial to eukaryotic biology. The Ub system relays an intricate control over cellular processes through its attachment and detachment on substrate protein, giving rise to a “ubiquitin code”. Deregulation in this code has severe consequences, primarily in disease pathogenesis. In contrast to other post-translational modifications being singular, ubiquitination comes in many different flavors. Subsequent attachment of the C-terminus of one Ub moiety onto a substrate lysine residue or one of eight amines (Met1, Lys6, Lys11, Lys27, Lys29, Lys33, Lys48, Lys63) of another Ub moiety gives rise to distinct Ub linkages that are recognized by the decoders or readers of the “ubiquitin code”. These Ub receptors facilitate important functions such as inflammatory response and DNA damage repair. Research over the past few decades has provided critical insight into the writers that establish and erasers that remove the “ubiquitin code”, yet our understanding of these Ub-receptors and Ub-interacting proteins is limited in comparison. This proposal describes the use of Ub-based probes for profiling UBDs and Ub-dependent protein-protein interactions using expanded protein chemistry. In two aims, we leverage genetic code expansion and liquid chromatography-tandem mass spectrometry to address the goal of this project: to develop a means to identify and structurally characterize Ub-dependent protein-protein interactions. Aim 1. Identify transiently interacting UBDs of eukaryotic origin using photoaffinity biotinylated Ub probes and structurally characterize promising candidates using chemo-selective ligation and X-ray crystallography. Aim 2. Develop a means to perform chemical (poly)ubiquitination using the traceless Staudinger ligation for access to homogenous (poly)Ub probes and (poly)ubiquitinated substrate protein. This will be applied to the atypical Lys29 ubiquitination of the 26S proteasome receptor Rpn13, a regulatory effect of the E3 ligase UBE3C. These approaches in Ub chemical biology will allow us to study Ub-dependent protein-protein interactions and their biophysical counterpart by reconstituting the elements needed to chemically trap noncovalent interactions, a feature currently limiting the study of the Ub system. Taken together, we seek to describe how these Ub-based interactions biochemically attenuate function and corroborate these findings using structural evidence.

NIH Research Projects · FY 2025 · 2022-07

PROJECT ABSTRACT This Mentored Research Scientist Career Development Award (K01) will support my pursuit of becoming an independently funded investigator in Alzheimer’s and aging research. My proposal features a systematic approach to a comprehensive training plan including mentorship, didactic and experiential learning activities, and professional development opportunities, as well as a complementary research project establishing foundational evidence for my research agenda centered on improving care of people with Alzheimer’s disease, related dementias, and diabetes mellitus (ADRD-DM). My training goals are designed to attain essential advanced knowledge and skills pertaining to 1) ADRD collaborative care and research, 2) DM collaborative care and research, and 3) health systems and clinical trials. My research and training will take advantage of the immense resources and expertise at supporting centers across Indiana University, the Regenstrief Institute, and Eskenazi Health, including the National Institute on Aging-funded Institute of Alzheimer’s Disease Research Center and the IMPACT Collaboratory, and will be conducted under the mentorship of a multidisciplinary team of ADRD, DM, and implementation science experts, who have clinical and research experience in creating innovative technologies to improve care delivery in ADRD-DM populations. Building on my expertise in human factors engineering, the research project will examine how use of information technology can increase awareness of hypoglycemia and shared decisions about treatment options among people with ADRD-DM and their caregivers and primary care clinicians. I will use a participatory design process to develop assistive information technology that leverages continuous glucose monitoring devices. In this experiential learning opportunity, I will gain insights into patients’ and caregivers’ perceptions and experiences, clinicians’ information gaps and other barriers to individualizing ADRD-DM treatment, and the design of technology to facilitate shared decision-making, while expanding the knowledge base on these aspects of health care. Furthermore, I intend to increase representation of diverse socioeconomical and cultural backgrounds among ADRD research participants and scientists. This K01 award will uniquely position me to bring together human factors and ADRD-DM research to design and implement technology to assess and improve the health of people with ADRD.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT Mimicking protein-like cavities and functions requires to precisely organize surface groups on well-defined, porous scaffolds to function in a concerted and orchestrated manner. Here, we propose to generate fully sequence-defined robust nanocages for selective, multivalent recognition and catalysis reminiscent of protein cavities. Building on the PI’s unique interdisciplinary expertise in supramolecular, organic, and computational chemistry, the primary objective is to create strategies to design and synthesize fully sequence-defined nanocages as artificial receptors for sensing biomolecules (e.g., enkephalins in the brain) and as selective artificial enzymes (e.g., for site-selective modification of peptides). We will direct the positioning of the endohedral functional groups (e.g., mono-, di-, or tripeptides) with chiral, sequence-defined covalent templates (e.g., sequence-defined peptide dendrimers and a-helical peptides), which will transfer their outer amino acids onto the endohedral sites of the nanocages in a stereocontrolled manner. The Schneebeli lab has synthesized and characterized robust, hydrazone-linked nanocages required to bulid the sequence-defined nanocages. These nanocages have been successfully applied as receptors for small molecules, and as size-selective polymerization catalysts, which lays the foundation for the proposed selective neuropeptide recognition and site- selective catalysis. We will pursue two parallel avenues to create the fully sequence defined nanocages. In Project 1, we will template nanocage formation directly in a sequence-defined manner, assembling the nanocages with hydrazone bonds (which are reversible under acidic conditions) and then cut out the templates. In Project 2, we will stereoselectively functionalize preformed [8+12]-nanocages (created with a novel, cyclotrimerization-based synthesis) with the help of a-helical peptide templates and equilibrating imine bonds, and then reduce the imine bonds and cut out the templates. This research is novel in the concept, synthetic approach, characterization, and applications, because (i) fully sequence-defined asymmetric nanocages represent a considerable shift in the current research of artificial molecular receptors and artificial enzymes, (ii) hydrazone-linked nanocages and cyclotrimerization-based nanocages syntheses (pioneered in the Schneebeli group) are powerful approaches to access large and chemically-robust nanocage frameworks capable of holding up to 12 endohedral functional groups with full sequence definition, and (iii) the creation of robust, protein-mimetic cavities may ultimately yield numerous advances in both fundamental structural and mechanistic knowledge of artificial receptor/artificial enzyme design. The proposed research is significant, because it will enable synthetic chemists to independently control the endohedral sites of porous nanocages, ultimately offering new opportunities and approaches to rapidly sense biomolecules and catalyze new, stereoselective chemical reactions for therapeutics development.

NIH Research Projects · FY 2025 · 2022-07

Project Summary Neutrophils play critical roles during different stages of tumor development. In mice, systemic depletion of neutrophils results in decreased tumor growth in glioblastoma (GBM) and lung cancer, but promote tumor growth in pre-metastatic lung and other solid tumors, indicating their stage-specific and tissue-dependent functions in tumor progression. Neutrophils could also facilitate cancer cell resistance to chemotherapy, radiotherapy, and immunotherapy in different tumors by releasing various cytokines. Despite these preclinical and animal studies on tumor-associated neutrophils (TANs), a knowledge gap remains in our mechanistic understanding of how human neutrophils regulate cancer progression and therapeutic resistance in GBM, due to the short life and resistance to gene editing of neutrophils as well as technical hurdles in isolating stage-specific TANs. To address this gap, we propose to harness the power of microfluidics and human induced pluripotent stem cells (hiPSCs) to interrogate the diversity and plasticity of neutrophils in human GBM development. Elucidating the underlying mechanism will also enable the much-needed development and evaluation of neutrophil-targeted cancer therapy. Our central hypothesis here is that the microfluidic model will recapitulate the different stages of human tumor progression, providing a platform for phenotypic and mechanistic understanding of the roles of neutrophils in GBM development. To test this hypothesis, we will implement a novel interstitial tumor-microenvironment-on- chip (iT-MOC), and interrogate neutrophil-mediated tumor progression and therapeutic resistance at different GBM growth stages in Aim 1. Then in Aim 2, we will determine the morphology, polarization, life-span and antitumor cytotoxicity of GBM-infiltrating neutrophils. In Aim 3, we will reprogram tumor-associated neutrophils towards antitumor effector cells via genetic engineering of hiPSCs with chimeric antigen receptors (CARs) and microRNAs (miRNAs). This is a novel approach as human neutrophils cannot be genetically modified. Successful completion of these aims will offer an innovative platform to study the diversity and plasticity of TANs, and provide insights into reprograming them towards antitumor effector cells and the proof-of-concept for CAR-neutrophils in targeted cancer therapy.

NIH Research Projects · FY 2026 · 2022-07

Project Summary Many cancers are attributed to chronic inflammation, which can cause mutations and activate oncogenic signaling pathways. An important example is gastric cancer, the fourth-leading cause of cancer death worldwide. At least 80% of gastric cancer cases are attributed to stomach infection with the bacterium Helicobacter pylori (Hp), which causes lifelong chronic inflammation that does not eradicate the infection. In some individuals, this inflammation can cause gastric atrophy, metaplasia (conversion of one normal cell type to another), dysplasia (presence of abnormal cells) and finally cancer, but the specific mechanism(s) through which Hp triggers this cascade are not well understood. Similar preneoplastic changes are recapitulated in a mouse model through tamoxifen-induced expression of active KRAS in the chief cells of the stomach (KRAS+ mice). I found that Hp infection of KRAS+ mice exacerbated disease: compared to Hp-KRAS+ mice, Hp+KRAS+ mice had an altered trajectory of metaplasia and accelerated dysplasia. Hp+KRAS+ mice also had expansion of “variant” pit cells (surface mucous cells) that expressed metaplasia- and cancer-related genes like the mucin Muc4. In accordance with the hypothesis that Hp causes cancer through eliciting chronic inflammation, Hp+KRAS+ mice had severe inflammation marked by a ten- fold increase in T cells vs. Hp-KRAS+ mice. In this proposal I will investigate the mechanism(s) through which Hp worsens disease in KRAS+ mice, with a broader goal of better understanding how Hp infection and its associated chronic inflammation cause cancer. I hypothesize that Hp modulates the inflammatory response to prevent Hp eradication from the stomach, and this deleterious immune response leads to metaplasia and dysplasia. In Aim 1 I will perform targeted depletion of CD4+ and CD8α+ T cell subsets to assess whether immune perturbation impacts metaplasia, dysplasia and Hp colonization. I will also assess whether T cells traffic to the stomach in Hp+KRAS+ mice or proliferate locally. Finally, I will test whether Hp+ human samples have increased T cells relative to Hp- samples. In Aim 2 I will investigate candidate Hp virulence factors that modulate inflammation: the cag type IV secretion system and two proteins that modulate T cells, the toxin VacA and the transpeptidase gGT. I will determine whether virulence factor mutants can elicit the same inflammatory and disease progression phenotypes in KRAS+ mice as wild-type Hp does. In Aim 3 I will test whether variant pit cells arise from gastric progenitor cells, and will assess disease phenotypes in Hp+KRAS+MUC4- mice to determine whether Muc4 expression drives pit cell transformation. I will also perform spatial single-cell RNA sequencing in +/- Hp, +/- KRAS mice to discover whether variant pit cells share a gene expression signature with Muc4-expressing cells in the “first gland” of the stomach, a morphologically distinct gland that is believed to be a source of reparative cell lineages. Taken together, the results of these studies will provide new understanding of how Hp infection and inflammation cause gastric preneoplastic progression and may reveal new targets for gastric immunotherapy. These findings may also be applicable to other cancers associated with bacterial infection and/or inflammation, like colorectal cancer.

NIH Research Projects · FY 2024 · 2022-07

The mission of the Office of Indiana State Chemist (OISC) is to protect the public consumer, livestock and pets, and manufacturers/producers from: Falsely represented and otherwise misrepresented feed, fertilizer, seed, and pesticide products in the marketplace; misuse of feed, fertilizer, seed or pesticides such that these may cause disease or harm to persons or animals, or unacceptably disturb the environment. This mission is accomplished by assuring truth in labeling for feed, fertilizer, seed, and pesticide products offered in Indiana commerce; by requiring training of identified users and handlers of these materials; and by regulating products and persons to gain compliance with Indiana and federal laws that have been assigned to OISC. OISC may involve legal remedies (fines and penalties) 1) to secure an expectation of safety in the purchase of feed, fertilizer, seed, and pesticide products marketed in Indiana for consumers, and therein, provide protection for pets or livestock; 2) to ensure responsible use of these same products through labeling and educating the user to gain compliance; and 3) to actively support an integrated food safety system and contribute to improving the Indiana environment. The primary objective of our proposal is to further develop, implement, and sustain Indiana's best practices for a quality regulatory program to enhance animal feed and food safety. This would entail the strengthening of interagency collaboration, cooperation, and communication; achievement and sustainment of compliance with the Animal Feed Regulatory Program Standards (AFRPS); enhancement of coordination and interactions with the U.S. Food and Drug Administration (FDA) and other states' feed inspection programs. Under this proposal we are requesting $300,000 annually in grant funds for the next five years. Grant funds will be used to augment current animal feed inspection program capabilities and provide the necessary infrastructure to continue further development and sustainment of AFRPS. Time of one full-time equivalent (FTE) employee, travel necessary for collaboration and innovation, and some laboratory equipment and supplies will enable enhancements to the program's ability to conduct high-quality inspections and investigations to reduce the incidence of animal feed contamination and ultimately foodborne illness associated adulterated animal feed material/supply facilities.

NIH Research Projects · FY 2025 · 2022-07

SUMMARY Industrial hygienists and other occupational safety and health (OSH) play a critical role in addressing occupational health hazards. However, there is an unmet need for highly trained OSH professionals; this need is even greater for industrial hygienists. Meanwhile, worker health is a substantial issue within Indiana, where workplace related injury and fatality rates are consistently higher than the national average. The Purdue University Occupational and Environmental Health Sciences (OEHS) program will address this need through training the next generation of industrial hygienists. This program includes PhD, MS, and BS degrees in OEHS; the BS and MS programs are ABET accredited in industrial hygiene. Since 2017, program faculty and students have received >$100 million in external funding, published more >35 peer reviewed journal articles, given >70 research presentations, and received >30 honors or awards in recognition of their research, teaching, and service. Faculty are noted for their excellence in both research and teaching within the areas of industrial hygiene, occupational safety and health, environmental health, exposure science, occupational toxicology, radiation health and safety, and occupational epidemiology. Required coursework for our programs includes safety and ethics, exposure assessment, physical agents, industrial hygiene measurements and instrumentation, safety engineering, ergonomics, ventilation, toxicology, epidemiology, environmental health, risk assessment and statistics. The core curriculum utilizes student activities that emphasize critical thinking, communications, teamwork, creativity and awareness of current issues. Recently developed courses additionally incorporate service-based or case-study based learning approaches. Program trainees have multiple opportunities to interact with industrial hygiene and OSH professionals at a networking events held in West Lafayette, Indianapolis, Chicago, and beyond. Faculty and students conduct award-winning research related to occupational exposure and impacts of manganese and other metals; industrial contamination leading to environmental disparities; characterization and risk assessment of aerosols; and radiation health and safety. The OEHS program, as well as Purdue University, value diversity, equity, and inclusion (DEI) and seeks to incorporate these principles throughout every aspect of the program. These efforts will be reflected in broadening recruitment efforts to reach faculty and trainees with a variety of backgrounds and experience; promotion of professional development programs and resources to enable every trainee to reach their full potential; and recognition that application of DEI values to the practice of industrial hygiene will be essential in order to address the hazards faced by an increasingly diverse workforce. The success of Purdue’s OEHS program is evidenced by the fact that program graduates have taken leadership roles in industrial hygiene and related fields. By combining past experience and adapting to future needs, the Purdue OEHS program will continue to train tomorrow’s leaders in the field of industrial hygiene and occupational health.

NIH Research Projects · FY 2025 · 2022-07

Abstract Non-technical (NT) skills encompass teamwork, communication, decision-making, leadership, and situational awareness and are critical aspects for safe care delivery in the operating room (OR). Non-technical skills of surgery have been identified as the most frequent causes for sentinel events that impact patient mortality, post- operative pain, and quality of life. Lapses in communication, teamwork, and leadership have been shown to account for 60% of major perioperative complications, most of which are related to communication failures. Given this strong link between NT and team skills and patient safety, several NT skills assessment tools have been validated specifically for surgical teams. However, they require the presence of trained observers or retrospective review of videos. Such methods are time-consuming and resource intensive to obtain. Importantly, these assessments are subject to multiple assessment biases of the observer which threatens their objectivity and value. Automated tools that do not require an expert observer to conduct real-time observations, do not need time-intensive audio-video analysis, and could provide objective, quantifiable, and continuous measurements of NT skill are, thus, highly needed and could allow for the more widespread implementation of NT skills training. The objectives of the current project are to 1) investigate and develop predictive models linking objective sensor- based data streams with individual and team non-technical skills, 2) test non-intrusive sensing devices in simulated surgical team simulation training for practicing professionals, and 3) test sensor's accuracy in distinguishing teams with and without TeamSTEPPS training. To achieve these objectives, three specific aims are proposed. Aim 1 will validate our multi-modal sensing system for measuring NT skills in surgical simulation. A collaborative team of human factors engineers and surgeon educators will perform summative usability evaluation and determine usability with the System Usability Scale and Technology Acceptance Model. For Aim 2, our team will implement sensors in well-validated surgical simulations with experienced and inexperienced teams. As sensors continuously collect data, observer-based non-technical skill assessment tools will be performed real-time and video recorded. Analytics, data fusion, and regression analysis will be performed to model sensor metrics with gold-standard ratings. Finally, we will evaluate whether sensors can distinguish differences in team skills between teams with/without TeamSTEPPS training. The expected deliverables will include usability validation of a non-intrusive sensing system and analytics framework for automated skill assessment that can be implemented in surgical simulation. If proven valid and acceptable through the proposed work, our objective NT skill assessments could be used in the future to assess effectiveness of team training and provide structured, individually-tailored feedback to enhance healthcare providers' NT skills. Thus, this work could ultimately reduce errors due to poor NT skills and enhance patient safety.

NIH Research Projects · FY 2026 · 2022-06

High throughput infrastructure for reaction screening and bioassays Mass spectrometry (MS) is a powerful and widely applicable analytical method for qualitative and quantitative analysis of compounds of all types and sizes. Desorption electrospray ionization (DESI) is an ambient ionization method in which samples are analyzed in the open air by impact of primary droplets. Given the ability to position an array of samples relative to the mass spectrometer, DESI-MS becomes a high throughput (HT) chemical analysis method. The power of MS as an analytical technique is well known but it is less commonly realized that MS can also serve as a preparative method, e.g. it can be used to deposit mass-selected ions on surfaces to create new materials in vacuo. Of significant interest to organic synthesis, a unique feature of DESI is that, upon impact, the spray of solvent used to analyze a reaction mixture generates secondary microdroplets in which reactions may be accelerated en route to the mass spectrometer. It is this remarkable feature that makes DESI- MS a powerful synthetic method combined with a built-in analytical capability. With support of DARPA, we built a high throughput system at Purdue capable of automated reaction screening at a rate greater than 1 reaction mixture per second. We now propose an intramural - extramural collaboration between Purdue and the NCATS ASPIRE laboratory. The UG3 component of the collaboration will focus on designing, fabricating, and testing an improved high throughput system for reaction screening based on DESI-MS. The system will replicate the capabilities of the existing Purdue system and also include new capabilities for small-scale synthesis combined with high throughput bioassays. In the UH3 phase of the proposed study, the system will be transferred to NCATS and used in collaboration with the intramural group. As an initial demonstration of the new high throughput platform capabilities, we will pursue the discovery of novel therapeutics for advanced-stage prostate cancer, for which current chemotherapeutic agents show limited effectiveness. Specifically, this effort will entail large-scale screening and synthesis of potential cholesterol sulfotransferase (SULT2B1b, a currently undrugged biological target) inhibitory compounds, together with late-stage functionalization of bioactive scaffolds to generate a diverse range of analogs. Through this effort, the system will be established as an all-in-one next-generation drug discovery platform, with integrated screening, synthesis, and biological assay capabilities. During the latter stages of the UH3 phase, the versatility of the system will be tested in several other biological applications, including directed evolution and functionalization of acetylcholinesterase reactivators. Successful completion of these tasks will demonstrate the newly constructed high throughput DESI-MS platform to be an efficient method for the discovery and expansion of chemical space towards currently undrugged biological targets.

NIH Research Projects · FY 2026 · 2022-05

PROJECT SUMMARY Lower-extremity peripheral artery disease (PAD) is a manifestation of systemic atherosclerosis that affects more than 236 million individuals worldwide. Patients with PAD have a worse quality of life (QOL) than their healthy counterparts, due in part to the marked decline in physical functioning. Few non-invasive therapies currently exist to improve functional performance and restore QOL in people with PAD. The goal of the proposed experiments is to determine the benefits of home-based leg heat therapy (HT) on lower-extremity functioning and QOL in patients with PAD. This novel approach consists of custom engineered trousers and a portable water pump. Hot water is circulated through the trousers, evenly heating the buttocks, thighs and calf. This system is safe and convenient for application in the home setting without supervision. We demonstrated that repeated exposure to leg HT using these customized water-circulating trousers enhanced muscle strength and capillarization and accelerated recovery following muscle damage in young, sedentary individuals. In a preclinical model of PAD, we reported that HT via repeated lower-body immersion in a water bath enhanced skeletal muscle mass and strength. Our first randomized sham-controlled trial in 32 patients with symptomatic PAD revealed that supervised leg HT elicited a clinically meaningful improvement in perceived physical functioning. Our recent R21-sponsored pilot study revealed that home-based, unsupervised leg HT is safe and well-tolerated by symptomatic PAD patients. Building upon these extensive preliminary data, we propose to conduct a randomized, double-blind, sham-controlled clinical trial in 110 patients with PAD to establish the effect of daily home-based leg HT for 12 weeks on functional capacity and QOL. Patients randomized to the leg HT group (n=55) will be asked to apply the treatment daily for 90 min using water-circulating trousers perfused with water heated to 42ºC. In the sham group (n=55), water at 33ºC will be circulated through the trousers. The primary study outcome is the change in 6-minute walk distance between baseline and the 12- week follow up. Secondary outcomes include changes in leg strength and fatigability, changes in the short physical performance battery score and changes in perceived QOL. In Aim #2, we will determine the tissue- level mechanisms by which HT affects muscle strength and walking performance. We will assess skeletal muscle morphology and blood flow using magnetic resonance imaging (MRI) and biopsies from the calf muscle. If the conclusions based on our preliminary data are substantiated, the proposed experiments will provide an evidence-based framework of feasibility and efficacy of a novel, straightforward approach to improve functional performance and QOL in patients with PAD. Given its accessibility, tolerability and ease of use, HT has the potential for rapid translation and application in the clinical setting, thereby opening new horizons for the non-invasive management of PAD. Bruno Roseguini, Ph.D.

NIH Research Projects · FY 2026 · 2022-05

Project Summary/Abstract The candidate is a medicinal chemist and veterinary pathologist in the Department of Pathobiology and Diagnostic Investigation at Michigan State University (MSU). Her research interests focus on the preparation of small molecules to abrogate the aggregation of hyperphosphorylated tau (p-tau) in Alzheimer’s disease (AD) and related tauopathies, while other groups have sought aggregation inhibitors using recombinant tau that was not post-translationally modified. This K08 application will provide Dr. Jessica Fortin with the support necessary to accomplish 5 goals: 1) to gain knowledge on the pharmaceutical properties of potent abrogating molecules of p-tau fibrillization (ADME: absorption, distribution, metabolism, and excretion); 2) to apply knowledge for optimization of chemical structures; 3) to advance skills in conducting PK/PD studies in mice; 4) to integrate a mouse model for proof of concept studies; 5) to develop an independent research program in the neuroscience area of drug discovery. To achieve these goals and foster expertise in drug discovery, neuroscience, and translational research, Dr. Fortin has assembled a multi-disciplinary mentoring team comprised of Dr. Richard Neubig (primary mentor), a leader in drug discovery and development, and 5 co-mentors and one collaborator: Dr. Edmund Ellsworth and Dr. Babak Borhan, pioneers in medicinal chemistry and organic chemistry; Dr. Min- Hao Kuo, a pioneer in molecular biology of p-tau aggregation inhibitors; and Dr. Scott Counts, Dr. David Morgan, and Dr. Nicholas Kanaan (collaborator), pioneers in the design of translational studies using animal model of tau deposition. MSU has strong programs in drug discovery in multiple Departments. This project will enable Dr. Fortin to build a drug discovery program to inhibit aggregation of p-tau and its associated neurotoxicity, and propel small molecules to preclinical studies in drug development and proof of concept studies using a mouse model of tauopathies. Notably, Aim 1 organizes lead optimization steps prior to the preclinical stage of our drug discovery program. Aim 1 will allow the best novel p-tau aggregation inhibitors to be propelled to a higher level of testing to obtain preclinical data using the following: solubility test, protein binding assay, Caco-2 cell culture, P-glycoprotein substrate and inhibition assay, microsomal stability assay, and a blood-brain barrier in vitro model. Aim 2 will evaluate PK/PD, safety, and brain permeability of 4 best compounds in the same mouse strain to be used in Aim 3. As a proof of concept in Aim 3, the two best novel molecules will be tested for long-term reduction of neurodegeneration in a transgenic mouse model (PS19 P301S) of tauopathies. The model harbors the same human tau isoform used in the screening program. This grant will provide the training and support for Dr. Fortin to be integrated into the MSU College of Veterinary Medicine, allowing for an additional R01 proposal for the discovery and development of small molecule therapeutics in AD and related tauopathies. This will facilitate Dr. Fortin’s development into an independent researcher and contributor in the neuroscience drug discovery community.

NIH Research Projects · FY 2025 · 2022-05

The proposed research addresses a key patient safety gap among children with medical complexity (CMC) transitioning between hospital and home—medication related harm. CMC are often on multiple and complex medication regimens and have intensive healthcare needs. As a result, they experience frequent transitions of care, across professional roles and care settings and during these transitions, CMC are at great risk for medication-related harm. Most transitional care interventions have been adopted from adult care settings and may not be adequate to address the unique needs of CMC and their family caregivers. Interventions that are grounded in the unique situations and complexities of such patients are needed to improve medication use safety and family experience of care. This proposal describes the PI’s plan to transition into an independent investigator by developing expertise in 1) participatory, human centered design to co-develop useful interventions, and 2) implementing and evaluating patient and family centered interventions. The proposal also describes a 4-year project for developing a prototype care transition medication safety intervention and generating early evidence on its usability and factors influencing its implementation. Aim 1: Patient journey mapping—will be used to elucidate medication safety risks and contexts during the hospital-to-home transition period. Journey maps will be used to capture and visually represent the medication use experience of families across professional settings and boundaries. The output of this aim will inform prototype design under Aim 2. Aim 2: Participatory, human centered design to co-design a prototype medication safety intervention. Through multiple, iterative co-design sessions involving healthcare workers and family caregivers, a team will design a prototype medication safety intervention informed by journey maps from Aim 1, current literature, and participant elicitations. The result will be a composite prototype ready for usability testing. Aim 3: Will test usability of prototype intervention with healthcare workers and family caregivers. Additionally, barriers and facilitators to prototype implementation will be explored. Taken together, all 3 aims will yield a final, refined prototype intervention designed to improve medication use safety during care transitions. Future work will involve a randomized controlled trial of the prototype medication safety intervention to formally evaluate its effectiveness in preventing or reducing medication related harm as well as improving the medication use experience of CMC and their family caregivers.

NIH Research Projects · FY 2026 · 2022-04

Project Summary / Abstract Individuals with metabolic syndrome (MetS) compose a significant and growing proportion of our U.S. and global population (> 25%). It has been established that the presence of chronic diseases, such as MetS, enhances and prolongs environmental exposure-induced inflammation. Individuals with MetS have demonstrated enhanced inflammation due to ambient particulate matter exposures of which a significant proportion is nano-sized. The mechanisms associated with this enhanced susceptibility represent a significant gap in our knowledge. Mounting data from the Shannahan laboratory suggests that dysregulation of inflammatory resolution contributes to the exacerbated toxicity and disease progression observed in MetS. Specifically, nanoparticle inhalation exposures induce a pulmonary inflammatory response that is exacerbated and extended due to MetS. This inflammatory response corresponds with suppression of specialized pro-resolving mediators that facilitate inflammatory resolution. Our data suggests following inhalation, nanoparticles gain unique biocoronas on their surface that enhance the pro-inflammatory response while inhibiting resolution signaling. Further, our preliminary data demonstrates MetS disrupts ω-3 fatty acid metabolism impairing resolution. This proposal examines the hypothesis that dysregulation of inflammatory resolution following nanoparticle exposure mediates the susceptibility observed in MetS by exacerbating inflammatory responses and facilitating development and progression of chronic disease. The hypothesis will be tested through the completion of three main goals: 1) Delineation of pulmonary nanoparticle-biocorona alterations throughout metabolic syndrome development and the inflammation signaling consequences; 2) Determination of inflammatory resolution and specialized pro- resolving mediator kinetics following nanoparticle exposure in MetS and healthy mouse models; 3) Elucidation of differential ω-3 fatty acid metabolism in MetS following nanoparticle exposure. These mechanisms represent potential key regulators that are dysregulated in MetS, facilitating exacerbated responses and also are potential targets of therapeutic interventions. Typically, research and treatment strategies addressing exposure-induced inflammation focus on suppression of pro-inflammatory pathways rather than elucidation and effective stimulation of resolution processes. Completion of the project will generate new knowledge required to understand distinct mechanisms of toxicity in prevalent and sensitive subpopulations such as MetS. Elucidation of these mechanisms will allow for new disease prevention and treatment strategies while also broadening public health protections to environmental exposures.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY A central role of estrogen is regulating hydration balance in the body. Decreases in body (systemic) hydration, cause systemic vocal fold dehydration. Systemic vocal fold dehydration alters the biology of the tissue. Nevertheless, the importance of estrogens in regulating these biological changes of systemic vocal fold dehydration remain unknown. We address this important clinical and theoretical issue by investigating the transcriptome, proteome, tissue morphology, and ultrasonic vocalization outcomes in a study design that carefully manipulates hormonal state and hydration state. We will also examine the effects of estrogen on the vascular geometry and blood flow to the larynx following the interaction of systemic dehydration with altered hormonal states. We use the rat animal model for these studies since this model allows for robust and rigorous control of hydration and hormonal levels. Because estrogen is important irrespective of biological sex, both male and female rats will be included. To examine the effects of hormonal loss and replacement, we will measure outcomes when reproductive organs have been surgically removed (ovariectomized and orchiectomized rats) and when estrogen replacement with estradiol is delivered. Systemic dehydration will be induced by a translational paradigm of water restriction for 5 days. Animals with free access to water will serve as controls for dehydration. Across 16 animal groups, our first goal is to quantify the effects of hormonal loss on the molecular and histologic signatures of vocal fold dehydration and the influence of estradiol replacement on these signatures. Because hormone receptors are differentially located across vocal fold mucosa and muscle, and these tissue layers have distinct cellular organization and biomechanical properties, the vocal fold mucosa and muscle are examined separately. Our second goal is to investigate the interaction of hormonal status and dehydration on ultrasonic vocalizations. Our third goal is to examine the influence of hormonal status and dehydration on vascular geometry, blood flow, and the physiologic consequences of these changes. By combining molecular, histological, and functional studies, and mechanistic investigations of hemodynamic modifications, this comprehensive proposal seeks to shed fundamental insight into the effects of estrogens on vocal fold dehydration biology. These data are needed to drive clinical recommendations pertaining to vocal fold physiology under conditions of altered hydration and hormonal states.

NIH Research Projects · FY 2026 · 2022-04

Dose analysis for translating animal based vibrational force study for accelerating orthodontic tooth movement to clinic ABSTRACT Controlled Differential Tooth Movement (CDTM) refers to the ability to move teeth to be displaced faster, or to minimize the movement of teeth to be stationary (i.e., anchorage teeth or teeth during the retention phase). CDTM is highly desired in common orthodontic treatments such as canine retraction, canine impaction, molar protraction, and space closure. Successful CDTM drastically shortens treatment time and reduces common side- effects such as root resorption and anchorage loss. Studies show that an intermittent vibration force (IVF) superimposed on orthodontic force accelerates tooth movement. Further, in the absence of orthodontic force, IVF strengthens bone mineral density of the alveolar bone. However, currently there is little evidence to facilitate optimal selection of stimulation level. Furthermore, lack of control on stimulation level on the target tooth inevitably results in inconsistent reporting of outcomes. The overarching goal of the proposed work is to enable CDTM in the clinic by transitioning from successful animal studies to clinical applications. Objectives of the proposed project include: 1) identifying optimal IVF stimulation level for accelerating orthodontic tooth movement in rats as well as associated side-effects; 2) verifying effects of IVF on bone strengthening resulting in tooth stabilization; and 3) determining the threshold that can be used to scale stimulation level up for larger species like dogs and humans. We hypothesize that: (H1) there is an optimal level of IVF that accelerates movement of targeted teeth without side-effects; (H2) the same IVF can strengthen the bone surrounding the tooth without orthodontic force and reduce relapse during retention; and (H3) stress in the periodontal ligament (PDL) can be used as the threshold to effectively scale up the stimulation level from rats to larger species for achieving accelerated tooth movement. These hypotheses will be tested through three specific aims. Aim 1: Determine the optimal level of IVF (OLIVF) stimulation superimposed on an orthodontic load system that accelerates tooth movement in a rat model (H1) and the associated biological responses. Aim 2: Determine the effects of OLIVF on the tooth without orthodontic force (H2). Aim 3: Scale up stimulation level for larger species including dogs and humans, by normalizing to stress in the PDL, and validate the theory on dogs (H3). A PDL stress threshold will be used as the criterion for scaling up IVF from rats to dogs in this proposed study, with an eye toward scaling up to humans in future studies. Thus, a novel method to ensure delivery of the specified IVF on each individual tooth in the clinic will also be tested. This comprehensive study will pave the way for clinical trials using this technology. Further, associated biomechanics and biological studies will elucidate the mechanism behind IVF based CDTM, which will further advance the field as well as methods for applying this technology.