University Of Illinois At Chicago

NIH Research Projects · FY 2026 · 2023-05

HIV-1 envelope (Env) glycoprotein (gp) 160 belongs to class I fusion proteins that are also expressed by other highly pathogenic human viruses including influenza A viruses (IAV), Ebola viruses (EBOV), and coronaviruses (CoV) such as SARS-CoV (SARS1), MERS, and SARS-CoV-2 (SARS2). They build spikes on the viral envelope that induce fusion of viral and cellular membranes to allow viruses to enter cells, which is essential to the viral infection. Class I fusion proteins are synthesized as a type I transmembrane (TM) polypeptide precursor in the endoplasmic reticulum (ER) and delivered to the Golgi apparatus for maturation. The Golgi contains glycosidases/glycosyltransferases for glycosylation and conserved oligomeric Golgi (COG) complex and other associated proteins such as soluble N-ethylmaleimide-sensitive factor attachment receptor (SNARE) proteins for trafficking. Inside the Golgi, high-mannose-type N-glycans are processed into complex- type and hybrid-type N-glycans after extensive mannose-trimming, and O-glycosylation also occurs. These precursors except for SARS1-spike (S) are further subjected to proteolytic cleavage by furin to complete the maturation process. When these steps are disrupted in the Golgi, no infectious particles are produced, leading to complete inhibition of viral infection. Recently, we and others reported that MARCH8, a member of the membrane-associated RING-CH-type E3 ubiquitin ligase family, broadly inhibits viral replication by targeting a wide range of fusion proteins. Importantly, we reported that MARC causes multiple defects in class I H8 fusion maturation in the Golgi via an unknown mechanism. These defects are found not only in furin-cleavage of HIV-1 gp160, IAV-hemagglutinin (HA), EBOV-glycoprotein (GP), MERS-S, and SARS2-S, but also in N- and O-glycosylation of SARS2-S, MERS-S, and EBOV-GP in the Golgi. Although MARCH8 does not trigger the degradation of these fusion proteins, its E3 ligase function is still required for causing these defects. The goal of this project is to elucidate the molecular mechanism of these multiple defects in HIV-1 gp160 maturation by understanding the MARCH8 antiviral mechanism. We hypothesize that MARCH8 targets glycosidases, glycosyltransferases, furin, COG complex, and/or SNARE to block HIV-1 gp160 maturation. We propose two distinct but inter-related Aims to test this hypothesis. In Aim 1, we will characterize how MARCH8 blocks gp160 maturation during HIV-1 infection. Experiments will be performed in primary cells and human T cell lines in combination with RNA silencing and CRISPR/Cas9 knockout to elucidate the MARCH8 anti- HIV activity. In Aim 2, we will identify the MARCH8 targets that play a critical role in HIV-1 gp160 maturation. We will focus on 18 Golgi proteins selected by high confidence bioinformatic analysis to identify the targets. The significance of this project is very high, which will not only fill in gaps in our understanding of class I fusion protein glycosylation and trafficking in the Golgi, but also elucidate a novel antiviral mechanism that can be broadly applied to several highly pathogenic human viruses including HIV-1, SARS2, EBOV, and IAV.

NIH Research Projects · FY 2025 · 2023-05

PROJECT SUMMARY This application proposes a customized research training plan designed to promote the development of the applicant into an independent investigator. The plan includes advanced training in both bioinformatics and laboratory experimentation, along with tailored professional and career development opportunities. The training plan is supported by the outstanding availability of local and institutional resources at UIC. The proposed research will examine the mechanisms that control scar formation, a common result of the healing response. In adults, the outcome of wound repair is almost always a fibrous scar composed of disorganized extracellular matrix (ECM). Although regulation of scar formation is complex, a key feature is fibroblast (FB) activation, which generates ECM and contractile forces. Scarring and fibrosis occurs in many tissues and can cause significant impairments of the organ system affected. Recent studies in our lab have identified a novel FB function in wounds that may be linked to scar formation. These studies show that wound FBs can act as non- professional phagocytes and ingest apoptotic cells. Following apoptotic cell engulfment, FBs develop a fibrotic phenotype with enhanced migration, increased contractility (α-SMA expression), and increased collagen synthesis. One factor found to be significantly upregulated in fibrotic phagocytic FBs is microfibril-associated protein 5 (MFAP5 or microfibril-associated glycoprotein 2/MAGP2). MFAP5 influences microfibril function and can regulate cell signaling pathways. Interestingly, MFAP5 has been linked to fibrosis in several human diseases, including some cancers and fibrotic diseases. Still, little is known regarding its role in wound healing and scar formation. Therefore, the goal of this study is to gain a better understanding of the function of MFAP5 in skin healing and scar formation. We hypothesize that upregulation of MFAP5 in the healing skin wound modifies the FB response and promotes scar formation. In this study, the role of MFAP5 in wound healing will be investigated in Mfap5-/- mice, a well-established Mfap5 knock out mouse line. The effect of MFAP5 on FB phenotype will be further investigated in vitro. This proposal will utilize in vivo and in vitro wound healing assays and advanced bioinformatics techniques. Aim 1 will assess how the loss of MFAP5 affects wound healing, including scar collagen content and architecture, wound breaking strength, myofibroblast activation, angiogenesis, and wound closure. Single-cell RNA sequencing will be used to identify the FB subpopulation that produces MFAP5 during wound healing. Aim 2 will examine how MFAP5 influences FB function and gene expression by utilizing in vitro wound healing assays as well as bulk RNA-sequencing and functional pathway analysis. Together, the Aims will lead to a better understanding of the importance of MFAP5 in healing skin wounds and scar formation. Information gained from the proposed research may lead to the development of novel therapeutics and/or discovery of a prognostic biomarker for treatment of fibrotic diseases.

NIH Research Projects · FY 2026 · 2023-05

ABSTRACT Hypoxia is one of the main features of solid tumors including breast cancer and has been shown to correlate with a poor prognosis. Although many chemotherapeutic agents such as paclitaxel and doxorubicin are significantly effective in a normoxia environment, they are less effective in hypoxic tumor regions due to poor infusion, hypoxia, and acidity. Given its significant impact on treatment and tumor progression, the development of a new approach to specifically target the hypoxic regions of tumors is clinically needed. To address the challenges (unmet medical needs), our objective for this proposal is to develop a new probiotic-assisted approach as bacteria preferentially migrate and accumulate in the hypoxic region of the tumor. We developed our original hypoxia-inducible expression system and have optimized its use in E. coli G3/10 cells, which have been used as a probiotic in humans. We demonstrated that the G3/10 cells preferentially localized the hypoxic regions of xenografted breast tumors in mice. With the new tools of the hypoxia-inducible expression system in G3/10 cells, we will deliver cancer-killing cupredoxin proteins, azurin or rusticyanin, in the hypoxic region of triple-negative breast cancer as they have limited treatment options and a worse prognosis. We hypothesize that the development of a new bacteria-assisted approach to express cancer-killing proteins under the hypoxia-inducible promoter will provide functional tools for specifically targeting hypoxic tumors. To test our hypothesis and improve current therapy, we designed experiments based on our “bi-directional” strategy; a combination of the G3/10-based therapy with standard chemotherapeutic agents (paclitaxel and doxorubicin), so that the chemotherapy suppresses tumors in the outer periphery regions, perfused tumor regions, and our bacterial approach kills tumor in the hypoxic inner regions. Our preliminary data strongly support this concept. Considering the NIH/NCI’s focusing areas in FOA PAR-22-085 (Microbial-based Cancer Imaging and Therapy - Bugs as Drugs), we specifically formed a multidisciplinary team that integrates expertise in basic, translational, and clinical breast cancer biology, microbiology, molecular biology, immunology, pathology, and biostatistics. With the five years of preclinical research proposed in this application, we will discern the value of engineered G3/10 cells as effective bacteria-assisted functional tools. Results from the proposed studies will potentially provide new and effective treatment strategies that specifically target the hypoxic tumor microenvironment.

NIH Research Projects · FY 2026 · 2023-05

Felder et al_R25_2022_Abstract_FINAL Modern engineers need to blend technical competence with global and competitive competence. For biomedical engineers, efforts to support these core competencies is to cultivate the ability to translate and commercialize design and innovation to impact the end-users. To meet this goal, we propose to establish a distributed and interdisciplinary pipeline for sustainable student-driven innovation. This proposal encompasses a comprehensive curriculum across disciplines to drive longitudinal project development and innovation. Our first specific aim is to enhance senior design project preparedness and student competency. This aim is met by thoroughly validating needs with a redesigned interdisciplinary clinical immersion program and by developing a new undergraduate biomedical engineering course to enhance student physical prototyping skills. Needs fully validated by biomedical engineering and Innovation Medicine program medical students with primary clinical experience ensures project development is aligned with the ability to commercialize a product, and enhanced student prototyping skills support realistic development with enhanced fidelity. Together these initiatives enable our second specific aim, which is to leverage interdisciplinary collaboration to enhance medtech device design. This aim is met by revising the undergraduate biomedical engineering senior design capstone to incorporate the same medical students from clinical immersion. Moreover, the senior design class will accept projects based on validated needs from clinical immersion, which enables accelerated pacing and inclusion of both verification and validation in class. Product development from senior design is then transitioned for further development to the same medical students with their own capstone experience. Continuing projects from clinical immersion to senior design and then to medical capstone has substantial benefit including the ability to retain technical development and pursuit of further development that was not otherwise possible (e.g., publication of development, execution of limited studies, pursuit of intellectual property) by one capstone experience alone. Effects of clinical immersion to validate needs and the prototyping class to enhance prototyping competency will be evaluated using surveys and deliverables. Effects of project origin (i.e., clinical immersion or other) and participation in the prototyping class will be evaluated on the ability of teams to effectively collaborate within and outside disciplines, design, verify performance requirements, and validate that design output meets original need. Long-term success of leveraging interdisciplinary collaboration for medtech device design will be evaluated by comparative assessment of medical capstone deliverables based on project origin and participation of the pipeline. This proposed pipeline has the potential to enable significant student-driven innovation. 1

NIH Research Projects · FY 2026 · 2023-04

PROJECT ABSTRACT Impairments in executive functioning (EF), cognitive processes that support self-regulation, disproportionately impact children living in poverty and increase vulnerability for childhood disruptive behavior disorders (DBDs; oppositional defiant disorder, conduct disorder), which trigger a cascade of mental health problems and psychosocial difficulties across the lifespan. Poverty-related stress and maladaptive parenting styles have been linked to alterations of neural and behavioral EF markers in children; despite this, no studies have studied if parenting prevention programs can directly target childhood EF, and through improving EF, reduce disruptive behaviors in at-risk children. This K23 Mentored Patient-Oriented Research Career Development Award application seeks to conduct a mechanistic randomized clinical trial to determine whether neural-behavioral indices of childhood EF is an experimental therapeutic target that can be modified via caregiver participation in the Chicago Parent Program (CPP; Aim 1). Consistent with the NIMH Research Domain Criteria framework, childhood EF will be assessed across multiple levels of analysis (electroencephalogram [EEG], behavior, survey). The project also seeks to evaluate whether increases in childhood neural-behavioral EF mediate the effects of CPP in reducing disruptive behavior problems over a short-term follow-up (Aim 2). The project will also explore whether increases in specific parenting practices (discipline, scaffolding), previously linked to individual differences in EF, mediate the effects of CPP in predicting change in childhood neural-behavioral EF (Aim 3). The sample will include 90 Medicaid eligible parent-child (ages 4-5 years old) dyads; and will employ a novel recruitment approach where the target child will have moderate-to-severe EF delays at baseline but does not meet diagnostic criteria for a DBD. Consistent with the candidate's translational clinical scientist career goals, the candidate seeks crucial training in: community-based research and dissemination and implementation science, advanced EEG methods to evaluate neural-behavioral EF targets of preventive interventions, and statistical skills to design and evaluate clinical trials. Training will occur in an outstanding interdisciplinary environment at the University of Illinois at Chicago, in the Department of Psychiatry and leverage the department's established strengths in community mental health services research, dissemination and implementation science, and affective neuroscience. Mentors Drs. Atkins, Fitzgerald and Bhaumik and consultants Drs. Klumpp, Wakschlag, Gross, and Handley have expertise in the training areas and extensive histories of early career mentorship, and will work together to ensure the candidate's successful transition to research independence. Study findings will support an R01 application to validate the results on a larger scale and launch the candidate's academic career as a preventive intervention scientist. Findings have significant potential to identify whether CPP, a cost-efficient parenting intervention, modifies neural-behavioral EF indices in urban poor children, a theorized etiological risk mechanism of DBDs, and therefore prevent DBDs.

NIH Research Projects · FY 2026 · 2023-04

Heart failure (HF) is one of the most frequent principal diagnoses for hospitalization and a leading cause of death in the United States. It has been estimated that up to 65% of HF hospitalizations are the results of insufficient HF self-care. Despite clear evidence that HF self-care reduces the risk of mortality and hospital readmissions, many HF patients struggle to take medications as prescribed, maintain a low sodium diet, monitor their weight and HF symptoms daily, and engage in physical activity. Existing HF self-care interventions delivered face-to-face or via telephone have had limited impact and reach because they require significant provider time and are not always accessible to patients due to socioeconomic constraints, geographic barriers, and other obstacles. There is a critical need for accessible and scalable interventions that are designed to assist patients with HF self-care while in the community. Consumer mobile health (mHealth) technologies (e.g., apps and sensor devices) hold promise for promoting HF self-care and expanding intervention delivery. However, their efficacy remains largely underexplored. To address this gap, our team developed a patient-centered HF self-care intervention (iCardia4HF) that combines the use of three commercially available mobile health apps and connected health devices (MyApps) with a program of individually tailored text messages (Text4HF) targeting modifiable behavioral factors to promote HF self-care adherence and improve clinical outcomes. We recently completed a NIH-funded pilot randomized controlled trial (RCT) to test the feasibility and preliminary efficacy of the iCardia4HF intervention over 3 months in patients with chronic HF. Results from this study provide important feasibility and preliminary data. The next step in our program of research and purpose of the proposed study is to conduct a fully powered, 2x2 factorial RCT to determine the independent and combined efficacy of the two iCardia4HF intervention components (MyApps and Text4HF) at 6 months, as well as their maintenance efficacy at 6 months post-intervention. A total of 360 HF patients with suboptimal adherence to HF self-care will be recruited and randomized to one of four conditions for 12 months: (1) Usual care, (2) Text4HF, (3) MyApps, or (4) MyApps&Text4HF. The primary outcome is percent of days lost due to cardiovascular hospitalization or death for any cause. Secondary outcomes are objective measures of HF self- care adherence (medication [MEMS], daily weighing and BP monitoring [Withings scale and BP monitor], low- sodium diet [urinary sodium], and physical activity [accelerometer]), self-reported HF self-care, HRQL, and major cardiac events (mortality, hospitalizations, ER visits). This study will provide important new knowledge that will critically shape our understanding about the potential of commercially available mHealth technologies and tailored TMs to improve HF self-care adherence and reduce hospital readmissions in patients with HF.

NIH Research Projects · FY 2026 · 2023-04

Abstract Osteochondral defects of the knee are common worldwide, yet there are few viable options for patients with damaged osteochondral tissue as current treatments do not consistently regenerate functional tissue. The standard of care for osteochondral defect repair is arthroscopic microfracture surgery, but this procedure often results in formation of mechanically inferior fibrocartilage formation. To overcome limitations of this and other surgical procedures, tissue engineering strategies, such as cell-laden biomaterial scaffolds, are promising alternative approaches to treat these defects. However, scaffold-based strategies face several challenges, such as interference with critical cell-cell interactions, potential immune and/or inflammatory reaction to the scaffold and its degradation byproducts, and unsynchronized scaffold degradation rate with that of new tissue formation. New cellular condensation strategies without a scaffold address these issues, however, it is still difficult to precisely control the architecture of the engineered tissues to mimic the sophisticated three-dimensional (3D) structure and organization of natural osteochondral tissues and their structure-derived functions. Recently, 3D bioprinting has been applied in tissue engineering with the potential to create complicated, high-resolution 3D structures. In addition, we have engineered the first technology capable of 3D printing a cell-only bioink and maintaining the printed structure, which is necessary to form cell condensations. The hypothesis of this proposal is that cellular condensation-based prevascularized osteochondral tissue constructs of precisely defined geometries can be directly assembled with human stem cells and endothelial cells via 3D bioprinting into a photocurable liquid-like solid, shear-thinning and rapid self-healing microgel slurry with spatially controlled presentation of tissue specific growth factors. Microgel photocrosslinking after printing will provide temporary mechanical stability for the printed constructs during culture to permit cellular condensation formation. This cell- only bioprinting strategy will be implemented to print seamlessly continuous two-phase osteochondral tissue constructs with a prevascularized bone phase and a cartilage phase. Specifically, this proposal aims to (1) determine the role of microgel properties on the resolution and fidelity of the cell-only 3D printed constructs, (2) engineer prevascularized osteochondral constructs with individual cell-only bioinks by spatiotemporally controlled delivery of vasculogenic, osteogenic and chondrogenic growth factors, and (3) determine the clinical potential of the 3D printed prevascularized osteochondral constructs by evaluation of new osteochondral tissue formation and integration with the host vascular networks and bone and cartilage repair in a full-thickness osteochondral rabbit defect model. This platform strategy has the potential to greatly enhance the lives of those suffering from osteochondral defects and may enable the engineering of other complex functional tissues in the body.

NIH Research Projects · FY 2025 · 2023-04

Necrotizing enterocolitis (NEC) is a devastating intestinal inflammatory disease that primarily affects premature infants and extremely low birth weight babies. Commonly observed risk factors for NEC are prematurity, formula feeding, intestinal dysbiosis, and infection. Previous studies strongly suggest a critical role of the inappropriate microbiome colonization and activation neonatal immune system in NEC development. However, the pathogenesis of NEC is elusive. Particularly, it remains unclear how the NEC-associated risk factors contribute to the disorder. In preliminary studies, we characterized the effect of formula-feeding on intestinal flora, gene expression, and immunobiology in neonatal mice. We also examined intestinal pathology of mouse pups which were fed with formula followed by induction of a particular antimicrobial immune response. We found that formula-feeding alone resulted in a distinct type of gut dysbiosis and pre-NEC intestinal molecular changes that predispose the neonatal gut to inappropriate microbiome colonization/infection and render intestinal mucosa to be a target of cytotoxic inflammatory cells. We further revealed that formula-fed but not dam-fed mouse pups developed NEC upon activation of a cytotoxic inflammatory cell-associated antimicrobial immune response. Thus, it appears that NEC develops following inappropriate microbiome colonization in premature infants as a consequence of “multi-hit pathophysiological events”. In this project, we will study new mechanistic insights into these events and determine how the interaction of multiple-hit events contributes to the development of NEC in two complementary aims: (1) We will first characterize the series of pathophysiological events that leads to NEC development, using a novel and pathologically relevant mouse pup model of NEC and up-to-dated in vivo experimental pathological and immunological approaches. Then, we will use RNAseq and cutting-edge bioinformatic analysis to delineate the transcriptomic response of the small intestine of mouse pups to multiple- hit challenges and to unravel the relevance of this novel mouse model of NEC for human NEC. Furthermore, we will study how major NEC risk factor-induced “multi-hit events” contribute to NEC development by focusing on inflammatory cells, mucosal inflammation-associated inflammatory mediators, and a unique signal axis that protects intestinal epithelial cells against inflammatory cell attack. We will achieve this aim using in vivo experimental approaches that draw on molecular biology and mouse genetic engineering techniques. (2) We will elucidate how formula feeding causes pre-NEC molecular changes in the small intestine of premature neonates by taking a multidisciplinary in vivo and in vitro approaches that incorporate techniques of organoid culture, molecular and cellular biology, microbiology, mouse genetic engineering and gnotobiogy. Together, our work will provide a novel mouse model relevant for human NEC, advance knowledge of how the interaction between NEC risk factors and activation of the neonatal immature immune system triggers NEC development, and gain mechanistic insights that will inform the development of new strategies for the prevention and treatment of NEC.

NIH Research Projects · FY 2026 · 2023-03

ABSTRACT Endothelial injury occurring during bacterial and viral infections results in uncontrolled accumulation of protein- rich fluid and inflammatory cells in the underlying tissue, hallmarks of acute lung injury (ALI), and acute respiratory distress syndrome (ARDS). Despite remarkable advances in supportive care, patient survival in the setting of ALI and ARDS remains near 40%. We have demonstrated a crucial role of transient receptor potential channel 6 (TRPC6) mediated Ca2+ entry in initiating inflammatory signaling that causes ALI. However, we also showed that mutation of isoleucine (I)111 for its isomer leucine (L)111 in TRPC6 or block of TRPC6 at isoleucine111 using a novel peptide allows the channel to gain new functions independent of Ca2+ entry for programming EC from inflammatory into the regenerative lineage, thereby resolving lung injury. Thus, understanding the mechanisms of action of I111 in inducing channel activity and the therapeutic value of blocking I111 to promote EC regeneration hold the key to preventing these diseases. We show that: 1) substitution of I111 for its isomer L111 in the Ist ankyrin repeat domain (ARD) of TRPC6 blocks Ca2+ entry; 2) I111L mutation initiates allosteric transitions in TRPC6 leading to loss of channel function, based on nuclear magnetic resonance (NMR) studies; 3) I111L-TRPC6 induces EC regenerative signaling during injury as evidenced by expression of ERG, a transcription factor maintaining EC homeostasis, and EC proliferation, leading to rapid lung repair after injury; 4) rescue of WT-TRPC6 but not the I111L-TRPC6 mutant in EC of Trpc6-/- mice reinstates LPS-induced lung vascular hyperpermeability by suppressing the expression of ERG but augmenting NFB-expression and inflammatory signaling; 5) inducing conditional deletion of ERG in EC impaired EC proliferation and induced lung injury, and, 6) a TRPC6 blocking peptide spanning I111-TRPC6 suppresses Ca2+ entry in EC but promotes EC proliferation and resolution of lung inflammatory injury. Epigenetic changes in chromatin accessibility enable signal- dependent activation of transcription factors that bind gene promoters and enhancers to dictate cell functions. Intriguingly, ATAC-seq and Chip-seq of EC sorted from control versus injured lungs suggest that WT or mutated channel selectively activates the EC epigenome either in favor of NFB or ERG transcriptional activities to switch EC phenotype, thereby dictating the outcome of lung injury. Based on these exciting findings, in Aim#1, we will determine the novel mechanisms induced by isoleucine111 in regulating TRPC6 structural organization and functions. In Aim#2, we will test the hypothesis that in contrast to WT-TRPC6, the I111L TRPC6 mutant gains new functions independent of channel activity to program the EC epigenome to adopt a regenerative lineage and therapeutically blocking this residue function will therefore repair the vascular injury in the pre-clinical models of lung injury. Studies will use multipronged approaches, including molecular modeling, multi-omics, and 2- photon imaging of lung EC, along with an I111-TRPC6 blocking peptide to accomplish these aims. We believe these studies to be translational for developing specific TRPC6 antagonists to prevent ARDS.

NIH Research Projects · FY 2024 · 2023-03

Discovery of a pigment produced by Streptococcus pyogenes Abstract This proposal seeks to advance a new observation that the human pathogen Streptococcus pyogenes (Group A Streptococcus, GAS) produces a pigment that has gone unreported. We hypothesize the pigment protects GAS from host-derived and/or environmental stressors (such as reactive oxygen species) and likely enhances its fitness and potentially augments its virulence. For 80 years, no reference in the literature exists of a pigment being produced by this important organism, and prior to that, only two citations allude to such a factor. We suspect this property has been overlooked because its presence only becomes apparent in the laboratory after culturing in a chemically defined medium, grown to high cell densities, and after exposure to oxygen. Understanding if and how the production of a pigment could boost the fitness of GAS could provide a new target that therapeutics or vaccines could take aim upon. The objectives of this proposal are to elucidate the composition and structure of the GAS pigment, to identify genes associated with its production and regulation, and to characterize its ability to protect GAS from stressors and immune cell killing while exploring any cytotoxic properties.

NIH Research Projects · FY 2024 · 2023-01

Project Summary/Abstract Mental health disorders are the most common disease of childhood. Yet, millions of teens do not receive mental health care. Most at risk are teens from underserved populations (e.g., low socioeconomic status; racial/ethnic and/or gender/sexual minority), who face a myriad of barriers to mental health screening and care. As such, traditional methods for reaching underserved teens with mental health disorders are not working, resulting in life-long health disparities and a significant public health impact. Consistent with the recommendations made in the NIMH National Advisory Mental Health Council Workgroup report, the goal of this K08 application is to use and adapt existing digital mental health technologies to advance the engagement, assessment, detection, treatment, and delivery of services for pediatric mental health. Specifically, the Accelerated Creation-to-Sustainment Model will guide the development and implementation of the Teen Assess, Check, and Heal (TeACH) System into a pediatric primary care clinic serving teens and families from the West Side of Chicago. In Aim 1, the PI and her mentorship team will collaborate with underserved teens (n=20) and their parents (n=20) to identify strategies to target top barriers to engagement as well as top ethical concerns and requirements for cultural relevance, usability, and usefulness of the TeACH System. In Aim 2, the plan for implementing the TeACH System will be refined through observations, interviews, and co-design workshops with pediatric primary care pediatricians and staff. In Aim 3, the TeACH System will be implemented into a primary care clinic and evaluated in a randomized trial for: 1) engagement and implementation outcomes; and 2) assessment of remediation of health disparities by analyzing differential outcomes (e.g., race, insurance status, individual perceptions of mental health) in a randomized trial. This innovative research will inform general digital mental health technology engagement adaptations needed for underserved teens and identify implementation practices to support the TeACH System in pediatric primary care settings. The PI and her mentorship team will also determine the feasibility and satisfaction of the TeACH System in preparation for the PI’s planned expansion of the System across multiple primary care clinics in a future R01 proposal. The proposed research and career development plans logically build from the PI’s foundational training in pediatrics/behavioral health, user-centered design, and mobile health (mHealth) evaluation to provide opportunities to gain knowledge and skills in: 1) pediatric health disparities; 2) dissemination and implementation science; and 3) ethics specific to deploying digital mental health technologies for underserved populations. Supported by an interdisciplinary team of experts and in institutional environment invested in supporting innovative initiatives to improve the mental and behavioral health of underserved populations, this K08 will launch the PI into a successful career as an independent clinical scientist.

NIH Research Projects · FY 2026 · 2023-01

Project Summary/Abstract Corneal neovascularization (NV) can be caused by severe corneal injury or infection and is a leading cause of blindness. Balance of pro-angiogenic factors and anti-angiogenic factors are important to maintain avascular corneal tissue. Pro-angiogenic factors such as VEGFA and FGF2 are highly induced in inflamed corneas and lead to activation of its receptor proteins such as VEGFR2 and FGFR2. Membrane-type 1 matrix metalloproteinase (MMP-14) is involved in remodeling of extracellular matrix (ECM) through its proteolytic activity. Recent studies have revealed that MMP-14 is a regulator of VEGFA/VEGFR2-mediated corneal NV via unique and selective cleavage of VEGFR1 which is a decoy receptor for VEGFR2. FGF2-induced corneal NV is delayed in MMP-14 knockout (KO) mice, indicating there is some correlation between FGF2 and MMP-14. However, how MMP-14 interacts with FGF2 is still largely unknown. The goal of this application is to characterize mechanism of MMP-14 on regulation of FGFR2 levels via ADAM-9 enzyme. We demonstrated that FGFR2 level was low in MMP-14 KO corneal fibroblast cells. On the other hand, ADAM-9, which is substrate of MMP-14, was higher in MMP-14 KO fibroblast than WT cells. We have also discovered that expression of MMP-8, MMP-9, and ADAM-17, all of which are underlying FGF2/FGFR2-system, were highly induced in WT than MMP-14 KO cells upon stimulation of FGF2. Thus, inhibition of MMP-14 can reduce FGF2/FGFR2-mediated corneal NV and inflammation. Furthermore, our results show that FDA-approved small molecule drugs, clioquinol, chloroxine, and folic acid, all of which contain a quinoline scaffold, inhibit MMP-14 enzyme activity. We propose three specific aims: (1) Mechanism of two enzyme cascades, MMP-14 and ADAM-9, to regulate FGFR2 level and expression of FGF2/FGFR2-mediated MMPs; (2) investigate the effect of MMP-14 in FGF2/FGFR2-mediated corneal inflammation; (3) Characterize quinoline analogs as selective MMP-14 inhibitors. We will complete these aims using innovative techniques from molecular biology and biophysics in vitro and in vivo.

NIH Research Projects · FY 2025 · 2023-01

ABSTRACT Hematopoietic stem cells (HSCs) are rare cells that reside in the bone marrow (BM) where they are maintained by specialized microenvironments (termed niches) in which endothelial, stromal, and other hematopoietic cells synthesize important niche factors that regulate HSC function. Mesenchymal stem cells (MSCs) are an essential component of the BM niche. These rare non-hematopoietic perivascular stromal cells are characterized by their unique ability to self-renew and differentiate into bone, cartilage, and fat, ensuring proper skeletal development and maintenance. BM MSCs form specialized niches that regulate HSC function by secreting high levels of niche factors such as CXC-chemokine ligand 12 (CXCL12), stem cell factor (SCF), and Vascular Cell Adhesion Molecule-1 (VCAM1). VCAM1 is classically expressed on endothelial and stromal cells where it acts as an adhesion molecule that preferentially binds to α4β1 integrin on HSCs and progenitors. Deletion of Vcam1 in endothelial and hematopoietic cells induces HSC and progenitor cell (HSPC) mobilization into the peripheral blood without affecting endothelial cell (EC) homeostasis. However, while the contribution of endothelial-derived VCAM1 to BM homeostasis has been extensively studied, the specific role of MSC-derived VCAM1 on HSC and on MSC maintenance and multilineage potency remains unknown. Our supporting data indicates that MSCs are the BM’s main source of Vcam1 and suggests that VCAM1 is critical for the maintenance, survival, and function of MSCs. Since MSCs are important regulators of HSC function and essential for skeleton and BM stroma formation and maintenance, it is critical to understand the extent at which MSC-derived Vcam1 deletion impacts MSCs and hematopoietic homeostasis. Based on our supporting data, I hypothesize that MSC-derived VCAM1 expression is critical for MSC maintenance and niche functions. The overall aims of this project are to elucidate the mechanisms by which VCAM1 promotes MSC survival and regulates HSPC function. Altogether, our proposal will provide mechanistic evidence for VCAM1 as a novel regulator of MSCs. While the critical regulators of HSC maintenance and differentiation have been intensively studied, that of BM niche MSCs still remain largely unknown. Our studies will not only aid in our understanding of MSCs but also the mechanisms encompassing HSC maintenance to ultimately help improve treatments for hematopoietic diseases.

NIH Research Projects · FY 2026 · 2022-12

SUMMARY: Despite decades of targeted research, no effective pharmacologic interventions have been identified for the Acute Respiratory Distress Syndrome (ARDS), which is a life-threatening disease process characterized by dysregulated immune responses. Sepsis is a major cause of ARDS, and the pathophysiology of both processes is characterized by alterations in microcirculatory blood flow, with vascular endothelial cell (EC) dysfunction playing a major role in organ injury. Novel mechanistic insights are needed to assist the development of therapies to address the EC barrier dysfunction that underlies ARDS and sepsis. Staphylococcus (S.) aureus is a frequently identified organism in gram-positive sepsis, and the highly virulent, antibiotic-resistant methicillin-resistant S. aureus (MRSA) strain is particularly challenging to treat and a major cause of ARDS. Important knowledge gaps exist both in MRSA-induced pathophysiology relevant to ARDS and in the mechanistic understanding of EC-specific processes that can be targeted therapeutically. Endogenous sphingosine-1-phosphate (S1P) and the structurally similar pharmaceutical compound, FTY720 (FTY), have EC barrier-enhancing effects in preclinical models of ARDS. However, both S1P and FTY also induce a myriad of other effects that are potentially harmful in critically ill patients and make them poor therapeutic candidates. We therefore have explored the barrier-regulatory properties of novel FTY720 analogs to better understand how these compounds regulate permeability. Our work has revealed that FTY720 (S)- phosphonate (Tys) has superior efficacy in several preclinical models and maintains expression levels of the essential S1PR1 receptor, unlike other agonists which induce its degradation. In addition, epigenetic processes are increasingly being recognized as important pathogenic steps during inflammatory acute lung injury (ALI)/ARDS and sepsis, and epigenetic responses (such as histone acetylation) can be altered by S1P-related signaling. Our central hypothesis is that MRSA causes EC dysfunction relevant to ARDS by epigenetic and other pathophysiologic mechanisms that can be targeted by Tys/S1PR1-related signaling. Using ChIP-seq analysis and other epigenetic tools, we have generated new insights that MRSA triggers histone acetylation in lung EC to regulate genes involved in lung EC dysfunction. Exciting new data suggest Tys-S1PR1 signaling may ameliorate key aspects of these epigenetic effects. Aim #1 will determine the mechanisms by which Tys/S1PR1 signaling protects against MRSA-induced lung EC barrier disruption in vitro. Aim #2 will use state- of-the-art ChIP-seq and other approaches to characterize novel MRSA- and Tys/S1PR1-induced epigenetic changes that have functional consequences in lung EC, including the novel MRSA target identified by our epigenetic screening, CYP1A1. Aim #3 will extend these studies in vivo by characterizing epigenetic and other mechanisms of MRSA-induced lung injury in mice and determine the efficacy of Tys. Overall, these studies will advance our mechanistic understanding of MRSA-induced ARDS to identify novel therapeutic targets.

NIH Research Projects · FY 2026 · 2022-12

Significant resources on age-related neurodegeneration are directed toward animal research in the assumption that results will inform our understanding of parallel processes in human. Yet, no reliable method exists to accurately translate cerebral blood flow or metabolic data from animal to human. For lack of rigorous mathematical methods for cerebral metabolic parameters between species, translation of valuable research data from mouse to human remains guesswork. There is a need for a predictive computational framework that quantifies cerebral blood flow and metabolism in normal and diseased human brain states. Our long term goal is to quantify fundamental physiological processes of aging and Alzheimer’s disease (AD), so that their effects can be slowed or even partially reversed. The objective is to expand the utility of MRI analysis by magnifying the detectability of age-related microcirculatory changes in humans with a mechanistic mathematical framework. It is our hypothesis that age and AD related changes in the microcirculation also generate macroscopic perturbations of hemodynamic and/or oxygen perfusion states that will be detectable with advanced MRI techniques, when guided by rigorous brain simulations over all relevant length scales. The rationale is that critical physiological metrics for dysfunction in aged brains (=aging biomarkers) will be exposed by systematic exploration and simulation of fundamental hemodynamic and metabolic processes. The central hypothesis will be tested by pursuing three specific aims: Aim 1) Assess the predictive value of mechanistic modeling by simulating the link between capillary stalling, vascular tracer transit properties, and tissue oxygen delivery, and validate predictions using advanced microscopic imaging in mouse. We determine the effects of aging and AD in aged rodent brains. Aim 2) Develop mechanistic multiscale model for predicting the impact of cerebral perfusion on oxygen metabolism in the human cortex under normal and pathological conditions. Anatomically detailed mechanistic models of cerebral circulation in human will predict the effect of structural and functional changes in AD on oxygen extraction in the human brain with MRI. Aim 3) Assess the predictive value of mechanistic multiscale modeling to quantify microvascular properties across the human brain cortex in health and disease using advanced MRI. To validate the mechanistic translation from mouse to human, we will measure age-related metabolic functions in cohorts of aged and Alzheimer patients. We identify hemodynamic and metabolic metrics (=biomarkers) that correlate with cognitive decline This contribution is significant because it will predict how changes in vascular morphometry and metabolism lead to neurological decline. It will identify biomarkers visible in noninvasive diagnostic imaging in humans that signal age-related deterioration before symptoms develop. The mechanistic framework relating data acquired in mouse to human will dramatically boost the relevance of animal data for human medicine.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY/ABSTRACT: Carbapenem-resistant Klebsiella pneumoniae (CR-Kp) cause life- threatening infections that are associated with unacceptably high mortality rates. CR-Kp is particularly challenging to treat since it often possesses a myriad of molecular resistance determinants that enable it to grow in the presence of most antibiotics. New β-lactam/β-lactamase inhibitors (BL/BLI), such as ceftazidime- avibactam, are not a sustainable therapeutic solution for CR-Kp as monotherapy since there is potential for development of resistance and clinical failure rates remain remarkably high when used alone. Up to 95% of CR- Kp still remain susceptible to at least one of the aminoglycosides (AMG). However, nearly all CR-Kp isolates harbor at least one of the aminoglycoside-modifying enzymes (AME), which inactivate a subset of the AMGs. Thus, selecting an AMG that is tailored to the AMEs and other AMG-resistance determinants (strain-specific AMG) is innovative and provides the foundation for this application, which is focused on development of molecularly precise AMG-based combinations for CR-Kp. Our central hypothesis is that novel combination regimens including short-courses of an optimally dosed, strain-specific AMG and a BL/BLI can maximize killing and resistance suppression of CR-Kp. Our promising preliminary data are highly supportive of our innovative and mechanistic approach. We developed a preliminary model to predict the strain-specific AMG based on an isolate’s AMG-resistance genes. We generated the first data for the combination of the strain-specific AMG with a BL/BLI (ceftazidime-avibactam, meropenem-vaborbactam, and imipenem-relebactam) against CR-Kp in both the hollow fiber infection model (HFIM) and the mouse pneumonia model, where the combination was highly synergistic. We have also developed a novel assay to quantify intracellular AMG concentrations. Leveraging our latest assays, we will elucidate the mechanisms responsible for synergy of the combination between a strain- specific AMG and BL/BLI in CR-Kp to rationally optimize them for future clinical trials. In Aim 1, the precise influence of each AMG-resistance determinant on AMG activity will be defined and novel predictors of AMG- resistance will be elucidated. In Aim 2, we will design novel short-course AMG treatment regimens that are efficacious. We will define the time-course of AMG-tolerance and resistance emergence using systems pharmacology and protein synthesis assays. In Aim 3, we will rationally optimize AMG and BL/BLI combinations by identifying mechanisms of synergy and assessing antibiotic target site concentrations. The HFIM will be used to define the pharmacodynamics of combinations and determine optimal timing of antibiotic administration. In Aim 4, we will develop quantitative and systems pharmacology (QSP) models that integrate our mechanistic data and rationally optimize AMG-based combination dosing strategies based on AMG-resistance determinants in CR-Kp. QSP-optimized combinations will be prospectively validated in mouse pneumonia models. This project will develop novel, molecularly precise AMG-based combinations to combat the urgent threat of CR-Kp.

NIH Research Projects · FY 2025 · 2022-09

Project Abstract Retinitis pigmentosa (RP) is the most common group of inherited retinal diseases (IRDs), leading to progressive photoreceptor degeneration and blindness. In addition to well-studied outer retinal dysfunction in RP, there is a growing body of rigorous work highlighting cellular adaptations in the inner retina, likely due to neural remodeling following deafferentation. The impact of this structural remodeling on the function of inner retina, and its consequences for visual function, are at present poorly understood. Electroretinography (ERG) provides a non-invasive method to assess inner retinal dysfunction both in the clinic and in animal models of retinitis pigmentosa, providing a means not only to provide insights into these questions, but also to translate mechanistic insights into potential therapeutic strategies for patients. The long term goal of the candidate is to become an independent clinician-scientist with the skills necessary to pursue strategies to prevent or restore vision loss in patients with IRDs. The scientific objective of the current proposal is to test the novel hypothesis, supported by preliminary data using a novel ERG protocol in both patients and animal models of retinal degeneration, that retinal remodeling leads to aberrant responses (noise) in inner retinal neurons that mask responses to a visual stimulus. The overarching goal of the proposal is to provide the candidate with the mentorship, skills, and support to realize a career dedicated to improve signal transmission in the degenerating retina. This is an essential area of research, as aberrant inner retinal responses limit the potential for functional improvement in all therapies that improve photoreceptor function. In Aim 1, the extent of inner-retina dysfunction will be assessed using novel ERG techniques in human subjects with RP. Aim 2 will develop ERG approaches to assess inner retinal remodeling in a mouse model of RP (the rd10 mouse model); Aim 3 will define the functional impact of photoreceptor degeneration in intracellular recordings of retinal ganglion cells in rd10 mice. This proposal describes a 4-year training program for developing an academic career focused on understanding retinal dysfunction in IRDs under an outstanding team of multidisciplinary mentors dedicated to the candidate’s objective of scientific independence. The proposal will leverage the candidate’s research training in neuroscience and clinical fellowship training in IRDs and vitreoretinal surgery under a mentorship team that has extensive experience in clinical retinal electrophysiology and psychophysics (Dr. Jason McAnany), animal electrophysiology and genetics (Dr. Neal Peachey), and single cell electrophysiology and imaging (Dr. Steve DeVries). Coursework and seminars will complement the mentorship team in the realization of ethical basic and translational research. The institutional environment in the Department of Ophthalmology & Visual Sciences at the University of Illinois-Chicago, with its corps of outstanding vision scientists and outstanding facilities, will maximize the candidate’s success in research and career development.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Out-of-hospital cardiac arrest is a significant public health burden, affecting over 350,000 people annually in the United States. Survival remains less than 12% and long-term neurological deficit is common in survivors. Currently no drugs exist to reverse myocardial stunning or improve long-term survival with good neurological outcome, which highlights an urgent and unmet need in resuscitation. Pharmacological stimulation of cardiac glucose oxidation represents a potential key strategy for enhancing recovery and reversing a pathological increase in fatty acid oxidation which occurs after ischemia, although agents such as dichloroacetate are limited by toxicity. A novel cell-penetrating peptide, TAT-PHLPP9c, was developed which rapidly gains access to tissues and selectively inhibits PHLPP1, thereby enhancing activation of Akt. In a mouse model of cardiac arrest, intravenous administration of TAT-PHLPP9c during CPR improves recovery of cerebral blood flow and enhances activation of both Akt and pyruvate dehydrogenase in the heart and brain. In swine, administration of TAT- PHLPP9c during mechanical CPR significantly improves 24-hour survival. Agents such as TAT-PHLPP9c that quickly gain access to tissues to improve survival would represent a significant advancement in resuscitation. Targeting metabolic dysfunction with TAT-PHLPP9c may decrease the circuit flow rate needed for successful eCPR, which will allow for narrow cannulas to be used and may lead to wider availability of eCPR. Using high resolution magnetic resonance methods, this project will investigate the mechanisms underlying TAT-PHLPP9c cardioprotection and neuroprotection. The following hypotheses will be tested: 1) TAT-PHLPP9c directly targets the heart and enhances functional recovery from ischemia; 2) TAT-PHLPP9c decreases fatty acid oxidation in the post-ischemic heart; 3) TAT-PHLPP9c treatment during low flow eCPR improves cardiac arrest survival, neurological recovery, cerebral blood flow, and cerebral metabolism; 4) TAT-PHLPP9c reduces the circuit flow rate required for successful eCPR; and 5) TAT-PHLPP9c-mediated Akt activation in peripheral blood leukocytes reflects Akt activation in tissues from vital organs. These hypotheses will be tested using a Langendorff model of rat heart ischemia/reperfusion injury, a swine model of eCPR, and a mouse model of cardiac arrest. It is our ultimate goal to translate these findings to emergency resuscitative care in order to save lives and minimize long- term neurological disability after cardiac arrest. A comprehensive training plan will develop the principal investigator’s skills as a physician-scientist under the guidance of the sponsor (Terry L. Vanden Hoek, MD, University of Illinois at Chicago) and co-sponsor (Henry R. Halperin, MD, MA, Johns Hopkins University).

NIH Research Projects · FY 2025 · 2022-09

Central retinal artery occlusion (CRAO) is an ophthalmological emergency with few proven therapies. Stem cell-based retinal cell replacement is a highly encouraging approach to achieve retinal neuroprotection and to save vision in retinal diseases. However, with limitations including few cells integrated, adverse immune re- sponses, and aberrant growth, an alternative cell-free approach is required. EVs are nano-vesicular bodies that, when endocytosed by target cells, trigger specific responses. Here, the microRNA (miRNA) cargo of the EVs plays a key role. This proposal targets restoration of retinal function using engineered MSC-EVs containing function-specific miRNA. Compared to MSCs, their EVs are non-immunogenic, non-tumorigenic, and modifiable for specific delivery modes. These characteristics render them ideal biomimetic agents fitting precision-based medicine. Our studies indicate that EVs can rescue retinal cells that have been acutely subjected to hypoxia or ischemia, the key mechanism that starts cells dying in CRAO. We also found that hypoxic preconditioning of MSCs resulted in EVs (H-EVs) with enhanced cytoprotective properties including anti-apoptosis and anti-inflam- mation. A number of miRNAs overexpressed in the H-EVs have cytoprotective properties in retinal cells. Our central hypothesis is that targeted EV-specific expression of miR-424/other key miRNAs in MSC-EVs will re-capitulate the anti-apoptosis and anti-inflammatory actions of H-EVs. We designate such EVs as Func- tionally Engineered EVs (FEEs). To facilitate clinical translation of MSC-EV therapy, we have identified key fun- damental knowledge gaps: (1) The relationship between EV miRNA and its anti-apoptotic properties; (2) EV miRNA and its role in anti-inflammatory actions of MSC-EVs in retina; and (3) Can MSC-EVs be enhanced for targeted functionality by engineering their miRNA cargo? In Aim 1 we will produce FEEs overexpressing miR- 424 (FEE-424) and 146b (FEE-146b). We will evaluate the mechanisms of action of the FEEs, and their candi- dacy for generation of FEEs in retinal ganglion cells, microglia, Muller cells, and retinal vascular endothelial cells using loss and gain of function studies in models of simulated ischemia in vitro. These results will serve as a proof-of-principle model for development of FEEs for amelioration of cell damage in the retina. In Aim 2, FEEs containing miR-424 and -146b will be used to test specific targeting of anti-apoptotic and inflammatory mecha- nisms in a rodent model of CRAO. Overall, the proposed studies are expected to provide transformative results whereby MSC-EVs are modified and delivered for retinal protective action after the ischemic event to treat CRAO. Innovations are cell-free therapy of retinal diseases, EV miR-mediated application-specificity, and direct determination of the impact of EVs on specific cells involved in retinal ischemic injury. Translational significance is the high likelihood of impacting novel molecular therapy. Underlying basic research significance is that the studies will enable vertical advancement of the field by determining mechanisms of actions of EVs in the retina.

NIH Research Projects · FY 2025 · 2022-09

SUMMARY Krabbe disease (KD) is caused by the deficiency of the ubiquitously expressed lysosomal enzyme galactosylceramidase (GALC) which is responsible for the degradation of galactosylceramides and galactosylsphingosine (psychosine). Because the synthetic pathway conducing to psychosine is not affected in KD, psychosine is continuously produced and accumulated in the Krabbe nervous system. Toxic levels of psychosine are considered the main pathogenic trigger of disease. Currently, the standard of care for KD is hematopoietic stem cell transplantation (HSCT), which is only applicable to asymptomatic or early symptomatic infantile KD cases and only protracts disease. Pre-clinical gene therapy studies using adeno-associated viral (AAV) vectors have shown great promise and in fact, AAV gene therapy applied early in life increases survival, improves quality of life, and decreases neuropathology in twitcher (twi) mice, the natural model for KD. Based on these important successes, AAV-based gene therapy clinical trials are being started only for infantile KD. However, despite the prevention of significant disease-related deficits, HSCT and pre-clinical AAV-gene therapy trials show varied long-term efficacy and resurgence of neurological disease. Thus, the status of gene therapy for KD, the limitations of HSCT to treat primarily presymptomatic infantile KD and the fact that juvenile and adult onset KD patients, which encompass a significant fraction of Krabbe patients, largely remain without any treatment, highlight the need to develop additional strategies to sustain long-term protection for KD patients. The use of substrate reduction therapies (SRT) strategies, singly or combined with current and new therapies for KD, is one potential way to achieve this. In this application we will use two small new compounds which selectively inhibit ceramide galactosyltransferase (CGT) and acid ceramidase (ACD), enzymes that mediate the production of psychosine via galactosylation of ceramides and sphingosine (CGT) and deacylation of galactosylceramide (ACD). Based on the premise that reducing psychosine synthesis will prevent/reduce psychosine-related pathology at early postnatal development of the mammalian brain, we will test the efficacy of SRT of CGT and ACD to enhance HSCT and AAV-GALC gene therapy in the mouse model of infantile KD (twitcher mouse) and the efficacy of single treatment with CGT or ACD inhibitors to ameliorate/prevent disease in a new model of adult-onset KD.

NIH Research Projects · FY 2025 · 2022-09

For dentin repair or regeneration, it is important to have the timely appearance of blood vessels. Therefore, tissue-engineering strategies to regenerate the dentin-pulp complex require establishment of vasculature to deliver oxygen, nutrients, hormones, immune cells, minerals and also help in clearing cellular debris and metabolic waste products during the inflammatory and regenerative phases of healing. DMP1 (dentin matrix protein1) is a key regulatory protein in bone and dentin mineralization. We first demonstrated that it has a regulatory role in the regulation of hydroxyapatite nucleation and growth in the extracellular matrices of bone and dentin. Subsequently, we demonstrated that DMP1 was localized in the nucleus of preosteoblasts and preodontoblasts and thus served as a signaling molecule and promoted the differentiation of these precursor cells. Recently we discovered that DMP1 can stimulate the release of intracellular calcium in preosteoblasts and preodontoblasts. Depletion of intracellular calcium from the endoplasmic reticulum leads to ER stress. Cells cope with ER stress by activating the “Unfolded protein response” (UPR). One of our recent observations is that DMP1 stimulation can promote the secretion of VEGF and other pro-angiogenic factors. Therefore, we hypothesize that ER stress activated by DMP1 functions to promote the transformation of adult stem cells such as dental pulp stem cells to endothelial cells and thereby promote vasculogenesis. In order to determine the mechanism by which DMP1 promotes vasculogenesis, we will examine the UPR signaling pathway. The UPR is initiated by three ER transmembrane proteins, of which our preliminary data show that DMP1 stimulation activated the ATF6 (Activating Transcription Factor 6) arm of the UPR. Accordingly, here we propose to study the mechanism by which ATF6 mediate transcriptional regulation of VEGF under ER stress. During dentin repair and regeneration, a major challenge is the maintenance of cell viability which depends on the availability of a functional vascular system. Accordingly, we will test the in-vivo vasculogenic competence and therapeutic potential of DMP1 in an in vivo pulp regeneration model. Understanding the complex functions of DMP1 could be valuable to develop therapies for fracture repair in bone or in the tooth to restore lost, damaged or diseased dentin-pulp complex.

NIH Research Projects · FY 2025 · 2022-09

Superwarfarins, also called long acting anticoagulant rodenticides (LAARs) are modified forms of the anti-coagulant warfarin, developed as potent rodenticides in the 1970's when rodents developed resistance to warfarin. LAARs are up to 100-fold more potent than warfarin and have extremely long half-lives (20 days or longer); one of the most commonly used is Brodifacoum (BDF). Increased use of LAARs led to an increase in accidental poisonings, mainly in young children. Those poisonings, which contain low amounts of BDF, are typically treated by providing plasma that contains clotting factors, and giving Vitamin K1 supplements for a few days. However, larger exposures occur due to unintentional (e.g. accidental spills) and intentional (e.g. suicide and homicide attempts) reasons; and LAARs have been used in terroristic and military attempts to cause injury and death on civilians and military, most recently by contamination of synthetic cannabinoids causing up to 400 hospitalizations and several deaths. While VK1 is used to prevent mortality from bleeding, it does not clear BDF from the body, so treatment can require up to a year at extremely high cost, nor does it reduce VK1- independent LAAR toxic actions which can lead to neuropathology and kidney damage. In previous studies we showed that treatment of BDF poisoned rabbits with cholestyramine (CSA), an FDA approved bile sequestrant which prevents enterohepatic recirculation, increased survival from 33% to 90%. This proposal expands upon those studies, with the overall goal to develop CSA as a countermeasure for LAAR poisoning to rapidly eliminate LAARs from the body. Studies will be done to optimize the amount and timing of CSA needed to increase elimination in adult rabbits, using different amounts of BDF as well as other LAARs; and the amounts needed to prevent the induction of kidney damage and neuropathology. Studies will be done to confirm CSA causes LAAR clearance in both male and female rabbits, and is able to prevent any consequences to neonates due to prenatal exposure of pregnant rabbits. Positive findings will provide the basis for eventual clinical testing of CSA in poisoned patients.

NIH Research Projects · FY 2025 · 2022-09

Project Summary. Vulnerable (Medicaid enrolled, low-income, minoritized, urban/rural resident) children are at high risk for severe early childhood caries (S-ECC). Treating young children for S-ECC often requires dental surgery under general anesthesia (DGA). Unfortunately, surgery is not an effective cure. A DGA event does not address underlying oral health behaviors such as tooth brushing and diet, so caries commonly recurs. Changing oral health behaviors is challenging, as parents struggle with concurrent life stressors. However, parents want to change their child’s oral health behaviors and identify that they need help to enact change at the time of a DGA event. We propose a behavioral parenting intervention to support oral health behavior change within families whose children experience the most severe disease burden at a critical point in their child’s oral health. We will develop and test the efficacy of PROTECT (Preventing Recurrent Operations Targeting Early Childhood Caries Treatment), a community health worker (CHW)-delivered, behavioral parenting intervention for preschool-aged children scheduled for DGA. The primary outcomes are frequency of tooth brushing and percent of total calorie intake from added sugar. The first session will occur in person at the time of surgery (60 minutes) and the remaining 10 sessions (30 minutes) will be delivered by phone to address barriers to engagement. The intervention will take place over a 6-month period, starting with the surgical event, with assessments measuring primary and secondary outcomes at baseline, 2 weeks, 6 months and 12-month follow up. Our investigators are experts in clinical (anesthesiology, dentistry, pediatrics, psychology, nutrition) and scientific realms (randomized controlled trials, management and analysis of longitudinal behavioral, psychosocial, and clinical outcomes, nutrition science, development of clinical interventions, dissemination and implementation). During the UG3 phase, we aim to develop PROTECT and test for feasibility and acceptability. The development of PROTECT will be informed by behavior change mechanisms of Social Cognitive Theory (SCT; e.g., positive parenting, self-efficacy, knowledge) and evidence-based behavioral parenting and dietary interventions, along with stakeholder (caregivers, providers, CHWs) input. We will identify barriers to recruitment, retention, intervention delivery, and outcome measurements. During the UH3 phase, we will test the efficacy of PROTECT compared to Usual Care (UC) to improve oral health behaviors. Participants will be randomized to receive PROTECT (n = 210) or UC (n = 210). We hypothesize that participants in the PROTECT group will increase tooth brushing and decrease added sugar intake to a greater degree than those in the UC group. Secondary outcomes and hypothesized SCT mechanisms of intervention effectiveness will also be tested. As an exploratory aim, saliva samples will be collected to assess changes to the oral microbiome from baseline to 6 months. This work fits within our broad research goal to improve children’s oral health, starting with the surgical population and potentially extending to children and adults within households.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT OF PROGRAM The loss of lung vascular barrier integrity in settings as diverse as trauma and bacterial or viral infections is a hallmark of acute lung injury (ALI) and its serious variant ARDS. ALI is characterized by protein-rich edema and ultimately respiratory failure. Targeted therapies remain an urgent unmet need. It is now becoming increasingly clear that the lung endothelium is a complex monolayer, almost an organ itself, consisting of not only alveolar endothelial cells (EC) but also specific EC populations found in pulmonary microvessels, arteries and veins. Recently, we have shown using RNA-sequencing that the lung EC demonstrate significant upregulation of genes involved in processes related to immune function such as leukocyte cell adhesion, leukocyte migration, and regulation of immune system. This finding was consistent with lung EC being continuously exposed to the external environment, unlike EC in other organs such as the brain or heart. Studying this immune regulatory function of the lung endothelium is crucial for understanding how the EC controls immunity and the host defense function of lungs, and also how its dysregulation or impairment of the immune response leads to pathogenesis of ALI. This Program builds on the extraordinary success of a previous 20-year entity, evident by our accomplishments. We have helped establish the lung endothelium as a node for understanding the lung’s response to infection and injury and our work has led to better understanding of ways of treating endothelial barrier breakdown in lungs. This revised application, focusing on the enigmatic innate immune function of the lung endothelium, is built on foundations of synergy and collaborations. Our Supporting data show the central role of the lung endothelium in driving inflammatory lung injury, and at the same time provides clues that will lead to new lung injury targeting therapies. Project 1 will test the hypothesis that the post-translationally modified endoplasmic reticulum-localized spinghosine-1-phosphate receptor S1PR1 in an unexpected manner reprograms lung endothelium to activate a signaling cascade that induces inflammatory lung injury. Project 2 will test the hypothesis that a novel lung endothelial cell expressed ubiquitin E3 ligase CHFR (checkpoint with fork-head and ring finger domain) identified by us regulates VE-cadherin-mediated endothelial barrier integrity and lung’s innate immune function. Targeting CHFR thus holds promise for preventing inflammatory lung injury. Project 3 will test the hypothesis that lung endothelial mitochondrial dysfunction and induction of mitophagy regulate endothelial regeneration and serve as a key check point for restoring homeostasis and preventing inflammatory injury. These Projects are supported by innovative scientific Cores (Epigenetics and Transcriptomics (Core B), Cellular Imaging (Core C), and Intravital Imaging and Physiology (Core D) that will make it possible to rigorously address the innate immune function of the lung endothelium and its role in orchestrating restoration of homeostasis. We hope to unravel the innate immune function of the lung endothelium, thus providing strategies to develop new targeted therapies against ALI and ARDS.

NIH Research Projects · FY 2025 · 2022-09

Summary Ebola virus disease (EVD) is caused by an infection with a group of viruses within the genus Ebolavirus. There are at least five species of Ebolavirus, Ebola virus (Zaire, EBOV), Ebola Sudan (SUDV), Bundibugyo virus (BDBV), Reston virus and Tai Forest virus (TAFV). Infections with these viruses can cause severe hemorrhagic fevers in humans and nonhuman primates, and are associated with up to 90% mortality rates with EBOV. Because of the safety concerns, these viruses are designated as the biosafety level 4 agents. Currently there is no effective therapeutic treatments against Ebola virus infection and pathogenesis in humans. Thus the goal of this application is to develop GP-specific small molecule inhibitors as drugs which can be used prophylactically and therapeutically against EBOV and other Ebola virus infections. To achieve this, we screened an in-house library of small molecules, and identified numerous potent entry inhibitors against EBOV. We have identified a series of potent compounds and will use them as leads which will be chemically optimized and developed as an anti-Ebola virus therapy candidate. In this application, two specific aims are proposed: (1) structure- based optimization of the lead compounds, and (2) In vivo evaluation of the lead compounds against EBOV infection using animal models.