Wayne State University

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY/ABSTRACT The hippocampus is composed of cytoarchitecturally-distinct subfields that support specific memory and learning functions. Evidence suggests distinct developmental trajectories in hippocampal subfields throughout childhood and adolescence, periods in which environmental factors exact strong influence on brain development. However, evidence is largely from cross-sectional samples, is based on inconsistent practices for delineations of subfields, and is limited by the power and diversity within single studies. Thus, developmental trajectories of hippocampal subfields remain unclear, prohibiting the systematic study of the influence of environmental factors on hippocampal development. Variations in total hippocampal volume across development have been linked to socioeconomic status, a proxy for access to material resources, medical care, and quality education in one’s environment. Low household socioeconomic status in childhood is associated with worse cognitive abilities later in adulthood, and low neighborhood socioeconomic status (an additional index of social capital and connectedness) further predicts poor behavioral outcomes beyond the household, especially in adolescence. The proposed study examines differential vulnerability of hippocampal subfield structures to variations in household and neighborhood socioeconomic across development. Understanding of normative developmental changes in subfield structure and vulnerability to low socioeconomic environments can have significant implications for individual and community interventions. However, testing these questions requires a large sample size, adequate coverage of age, specialized ultra-high-resolution scans, and sufficient representation of the diverse United States population. The proposed study will achieve this using integrative data analysis, an advanced latent modeling approach that directly addresses differences in methodologies while allowing new hypotheses to be tested in existing data. Hypotheses will be tested using a novel integrated longitudinal dataset of 678 typically developing subjects covering the span of 4- to- 25-years from four geographically- and demographically-diverse sites across the United States by pursuing these specific aims: Aim 1) Characterize typical developmental trajectories of hippocampal subfields in a diverse multi-site study pediatric sample; and Aim 2) Elucidate the link between variations in socioeconomic status and the development of hippocampal subfields. The hippocampus is implicated in neurodevelopmental disorders such as depression, schizophrenia, and low SES also confers risk for neurodevelopmental disorders. Thus, obtaining accurate characterization of hippocampal subfield development trajectories and identifying the effects of household and neighborhood SES in a large sample will pave the way for early identification of neurodevelopmental disorders and targeted intervention for specific factors associated with socioeconomic disparity. Further, the successful application of integrative data analysis to neuroimaging data from multiple studies will demonstrate a means to leverage existing data to answer critical questions otherwise not feasible in individual studies.

NIH Research Projects · FY 2025 · 2022-07

ABSTRACT Retinopathy is one of the most-feared complications of diabetes. In the pathogenesis of this blinding disease, retinal mitochondria become dysfunctional, the electron transport chain (ETC) is compromised, superoxide levels are elevated, and while complex III activity is inhibited, complex I remains unchanged. Mitochondria have their own small DNA (mtDNA), which lacks protective histones, but is packaged into nucleoids that provide some protection and assist in its biogenesis. Diabetes damages mtDNA, impairs its biogenesis, and downregulates gene expression of mtDNA-encoded cytochrome B (CYTB of complex III). Gene expression is also regulated by long noncoding RNAs (LncRNAs), the RNAs with >200 nucleotides and no open reading frame for translation, but they can bind to DNA or RNA, or can act as scaffolds to promote the interaction of proteins. Although majority of the LncRNAs are encoded by nuclear DNA, mtDNA also encodes three LncRNAs, LncND5 and LncND6 for complex I and LncCytB for complex III. Preliminary data show that in hyperglycemic milieu, while LncCytB is downregulated, LncND5 and LncND6 remain unchanged, and nucleoids are decreased and mtDNA sensitivity to nuclease digestion is increased. Based on these, our central hypothesis is that `LncCytB downregulation in diabetes impairs mtDNA nucleoids and attenuates cytochrome B transcription, damaging the mtDNA and the electron transport chain system, and the damaged mitochondria lead to the development of retinopathy'. Aim 1 will investigate the role of LncCytB in nucleoid formation, and the hypothesis predicts that `decrease in LncCytB in diabetes impairs nucleoids, damaging mtDNA integrity and reducing its copy numbers'. Aim 2 will examine the role of LncCytB in the regulation of the ETC, and will test the hypothesis that `downregulation of LncCytB decreases transcription of CYTB, which inhibits the complex III activity and compromises the ETC system'. Aim 3 will investigate the mechanism by which hyperglycemia downregulates LnCytB, and will examine the role of mitochondrial-targeted RNAse P protein 1 in regulation of LncCytB in the mitochondria. The plan will employ in vitro (human retinal endothelial cells) and in vivo (retinal microvessels from rodents) models of diabetic retinopathy, and will utilize fully optimized molecular biological approaches. Our overall goal is to identify novel regulatory mechanisms involved in the pathogenesis of diabetic retinopathy, specifically at the level of mtDNA-encoded LncRNA in mitochondrial homeostasis. The testable central hypothesis is innovative, and has significant translational impact as successful completion of our studies will provide strong background for LncCytB as a potential therapeutic target to prevent the development/ progression of this sight- threatening disease.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY The success of vaccination requires the generation of a strong immune response to the inoculated antigens in order to provide long-term protective immunity against many infectious diseases. To achieve this goal often requires the addition of vaccine adjuvants, substances that a substance that boosts the body’s immune response to the vaccine. However, there are only a few human vaccine adjuvants with an extensive safety record and minimal toxicity approved for clinical use. At presence, more studies are needed to identify novel adjuvants that not only significantly enhance the immune response for a particular vaccine, but also must be minimally toxic and maximally safe for clinical use. In efforts to discover novel vaccine adjuvants, an in vivo screening of forty-seven saponins from medicinal plants for their immunostimulatory and hemolytic activities has led to the discovery of new exciting vaccine adjuvants. Among forty- seven saponins evaluated, soyasaponins have emerged as the most potent adjuvants. These newly-discovered carbohydrates exhibited a significantly enhanced adjuvant activity with almost negligible toxicity when directly compared to QS-21 which has emerged as a vaccine adjuvant in numerous clinical trials. However, obtaining them from natural sources is a complicated process of extraction and purification that result in the production of minute. As a result, isolation of soyasaponins is economically unfeasible and unsustainable if sufficient quantities are required for immunological studies and clinical applications. Since FDA has strict regulations regarding to the purity and quality of adjuvants for use in human, a synthetic source must be developed for soyasaponins to be utilized as clinically relevant adjuvants. The objective of this proposal will address these challenges through the chemical synthesis for procuring sufficient quantities of soyasaponins in pure form. This effort will deliver well-defined soyasaponins without batch-to- batch variation and provide tools for studies of their roles as vaccine adjuvants and exploration of structure-adjuvant potency profiles for the discovery of non-natural soyasaponin improved adjuvants.

NIH Research Projects · FY 2026 · 2022-04

Significant progress in diabetes device technology has been realized over the past two decades. These novel technologies improve glycemic control over daily injections thus reducing the probability of encountering diabetic complications. Insulin infusion pump sets provide dosing flexibility and enhanced clinical efficacy in terms of reducing HbA1c and severe hypoglycemic events. Despite these technological improvements in insulin delivery systems, current best-practice set wear is typically limited to three days. Current challenges to extending the lifespan of subcutaneous insulin administration sets and infusion pumps involve unreliable insulin efficacy through the development of skin pathologies. Currently, all commercially available insulin formulations contain insulin phenolic preservatives (IPP) known as excipients that are a double edge sword. While they provide insulin protein stability, sterility and prolong insulin shelf life, our laboratory has recently shown that these are cytotoxic, induce inflammation and secondary fibrosis. Subsequently, our data in murine and porcine models demonstrated that proximate pre-infusion IPP removal significantly reduces infusion site inflammation while maintaining protein functionality. Thus, the two major obstacles to increased infusion set wear time are the chemotoxicity of the IPP and the transdermal cannula induced tissue injury, both of which are inflammation driven. Mature mast cells (MC) reside in cutaneous tissue. Thus, MC are one of the first responder in skin injury and are key contributors in orchestrating the inflammatory response once the skin is breached. Therefore, our central hypothesis, supported by our published and preliminary data, is that accumulative IPP and the transdermal injection and infusion devices contribute to local skin irritation due to mast cell activation and subsequent leukocyte recruitment, thus initiating the inflammatory cascade. As MC interact with macrophages (MQ) we further hypothesize that increased MC degranulation promotes M1 phenotype leading to phagocytosis insulin uptake/degradation by neutrophils & MQ and thus altering blood glucose control. Therefore, our overall goals are, first, to determine how MC activation occurs, and, second, the contribution to the resulting tissue reactions (inflammation and fibrotic cascades) while correlating IPP concentration and composition for the duration of the infusion period. We will test our hypothesis in three specific aims: 1) determine IPP induced MC activation and insulin degradation, 2) employ novel transgenic mouse models (Cre/loxP) to determine the mechanisms and mediators of IPP and device MC induced inflammation, and 3) preserve long-term tissue integrity during insulin infusion pump therapy in a pre-clinical porcine model. Ultimately, the successful accomplishment of this proposal could result in transforming current diabetes management practices that would achieve the goals of increasing the lifespan of insulin infusion devices and most importantly, sustaining a tissue site available for future recurrent insulin administrations.

NIH Research Projects · FY 2026 · 2022-04

Summary Intraflagellar transport (IFT) is an evolutionarily conserved mechanism for cilia formation. Defects in IFT/cilia have been linked to cilia-related diseases. Although the roles of IFT in somatic tissues have been extensively studied, little is known about its role in sperm flagella formation, which are specialized motile cilia with accessory structures. Using conditional knockout (cKO) strategies, our laboratory analyzed male germ cell- specific IFT mutant mice and discovered that all the analyzed IFTs are required for normal sperm formation/function. Among the Ift genes, Ift25 and Ift27 hold particular interest. The two IFTs form a heterodimer through their unique characterized structures. Although these two genes are not required for cilia assembly in somatic cells, both are essential for sperm formation and function. Specific elimination of each of these genes in male germ cells resulted in almost identical sterile phenotypes. Sperm from these mice were immotile and had disorganized accessory structures, especially the fibrous sheath. Levels of testicular pro- AKAP4, the precursor protein of AKAP4, an A-kinase anchor protein (AKAP) and significant component of the sperm fibrous sheath, were increased; on the contrary, the mature AKAP4 was significantly reduced in both Ift25 and Ift27 cKO mice. IFT25 associates with dynactin 4 (DCTN4), a dynein-associated protein. In addition to IFT25, IFT27 also associates with signal peptide peptidases like 2a (SPPL2a), which functions as a protease and is present in developing sperm flagella. The formation of mature AKAP4 was also affected in the Sppl2a KO mice. Based on these observations, we propose the following central hypotheses: 1 ) IFT25 and IFT27 are dedicated to the movement and placement of accessory structure components critical for functional sperm, and 2) The IFT25/IFT27 complex use specific domains to form IFT complex particles for sperm flagella assembling. To test these hypotheses, we propose the following Specific Aims: 1. To characterize the IFT25/IFT27 complex components in the testis essential for normal sperm morphogenesis, particularly the formation of accessory structures; 2. To investigate sperm accessory structure defects of Ift25 cKO mice dynamically and develop an in vivo system to track the IFT25 complex trafficking in live germ cells for sperm flagella assembly; and 3. To explore functional consequences of IFT25/27 disruption in sperm signaling. We propose that the IFT25/IFT27 heterodimer forms a transporting complex containing SPPL2a and DCTN4 through specific domains in male germ cells for normal sperm accessory structure assembly; we expect defects in accessory structures in the Ift25 cKO mice will occur at specific developmental steps. The dynamic trafficking process of the IFT25 complex in live male germ cells can be tracked. We hypothesize that SPPL2a is involved in processing pro-AKAP4 to mature into AKAP4, resulting in normal PKA signaling in mature sperm. The proposed research will elucidate the mechanisms of the two IFT proteins in the formation of functional sperm and build a platform to study the roles of other IFT components in male and female reproduction. .

NIH Research Projects · FY 2025 · 2022-04

Project Summary Dysregulated energy metabolism is intrinsically linked to the development of metabolic disorders, such as non-alcoholic steatohepatitis (NASH) and type 2 diabetes mellitus (T2DM). As universal electron carriers, both nicotinamide adenine dinucleotide (NAD) and its phosphorylated form (NADP) play essential roles in energy metabolism. The NAD kinase (NADK), which phosphorylates NAD to generate NADP, is exquisitely sensitive to nutritional or stress signals. While the cytosolic NADK has been characterized, little is known about generation and maintenance of NADP from mitochondria, the central organelle responsible for metabolic process and energy production, until our recent discovery that the uncharacterized human gene C5ORF33 encodes the long- sought mitochondrial NADK, referred to as MNADK. We have shown that MNADK, as a nutritionally-regulated, liver-enriched mitochondrial NADK, functions as the only de novo mitochondrial NADP biosynthesis pathway. Recently, we accumulated strong preliminary evidence that MNADK is required to maintain energy homeostasis, redox state, and insulin sensitivity and that repression of MNADK activity by protein S-nitrosylation modification under the high-fat diet is largely responsible for hepatic steatosis and insulin resistance induced by overnutrition. MNADK functions as a key regulator of acetylation of major metabolic transcriptional regulators. These observations lead us to hypothesize that MNADK is required to maintain energy homeostasis and insulin sensitivity and that repression of MNADK activity by overnutrition critically contributes to the development of NASH and T2DM. Mechanically, MNADK not only plays major roles in maintaining mitochondrial function and anti-oxidative protection, but also functions as a key regulator of acetylation of major mitochondrial regulators or enzymes that shape metabolic adaptability following metabolic challenges. In this application, we will utilize molecular and cellular approaches, genetically-engineered mouse models, as well as bioinformatics to critically address the functions and mechanisms by which the sole mitochondrial NAD kinase, MNADK, maintains energy homeostasis and protects from metabolic disorders. Our goal will be achieved by two aims: Aim 1, to determine the functional significance of MNADK in preserving hepatic energy homeostasis and thus mitigating the risk of metabolic disorders; and Aim 2, to decipher the molecular mechanism by which MNADK maintains mitochondrial function and energy metabolism. Upon completion of this project, we will have defined the function and mechanism by which the mitochondria NAD kinase preserves energy homeostasis and thus mitigates the risk of metabolic disease. Our proposed research will not only open a new paradigm for the study on molecular basis underlying energy metabolism, but also have important implications in the prevention and treatment of metabolic disease.

NIH Research Projects · FY 2026 · 2022-04

ABSTRACT Benzene is a prominent volatile organic compound (VOCs) that is present in water, food, paint, detergents, vehicle exhaust, tobacco smoke, and e-cigarette vapors. Exposure to low doses of environmental benzene in urban areas has been implicated in increasing the risk for metabolic dysfunction across all ages. However, a direct link between exposure to low-dose benzene and metabolic homeostasis is not yet established. Using the limited available literature on environmental exposure to benzene and its metabolic outcomes, we performed a preliminary meta-analysis and found a positive association between exposure to benzene and metabolic impairments. Our preliminary studies provide strong evidence that chronic exposure to benzene at varying low doses, modeling human exposure routes, induces significant insulin resistance and hyperglycemia in rodents. Neuroinflammation is increasingly recognized as one of the causal factors in the pathology of metabolic diseases. Glial cells (microglia and astrocytes) have recently garnered specific attention for their role in neuroinflammatory responses in metabolic disorders. Microglia, produce various pro-inflammatory molecules that are critical for the development of peripheral metabolic imbalance and insulin resistance via hypothalamic inflammation. We show that benzene exposure at several low doses relevant to occupational and environmental exposure promotes robust hypothalamic glial activation and elevation in the hypothalamic inflammatory IKKβ/NF-κB signaling pathway followed by the induction of endoplasmic reticulum (ER) stress response. Our central hypothesis is that benzene-induced changes in microglial function and IKKβ/NF-κB signaling underlie changes in whole-body glucose homeostasis and metabolic responses. This hypothesis will be assessed with a novel murine model of air-pollution combining molecular, genetic, and physiological approaches designed to manipulate both the number and the inflammatory activation state of resident microglia in the following Specific Aims: 1) To determine the contribution of exposure to low benzene concentrations to neuroinflammation and metabolic regulation; 2) To determine the role of central IKKβ/NF-κB inflammatory mechanism in a benzene-induced metabolic imbalance: 3) To determine the cellular and molecular interplay between microglia neuroinflammation and ER stress response triggered by benzene exposure. The proposed research will, for the first time, directly assess the role of benzene-induced changes in glial function and inflammatory signaling in regulating whole-body metabolism. Such a study will be of importance for shaping public health policy regarding benzene exposure and its role in predisposition to develop metabolic diseases.

NIH Research Projects · FY 2025 · 2022-04

Program Director/Principal Investigator (Last, First, Middle): Rossi, Noreen F. Nearly half of US adults have hypertension. Fructose intake predisposes to salt-sensitive hypertension, an independent risk factor for major adverse cardiovascular events (MACE) and chronic kidney disease (CKD). Increased salt intake impairs vascular compliance even before frank hypertension develops. Aortic stiffness is now recognized as a robust predictor of MACE and CKD. Sympathoexcitation increases cardiovascular risk and strongly impacts aortic stiffness. Our preliminary data show that combined fructose and salt intake contributes to insulin resistance and hypertension that displays increased renal sympathetic activity and aortic as well as renal artery stiffness. The goal of this application is to achieve early identification and timely intervention of vascular stiffness to mitigate MACE and CKD. Robust, rigorous preclinical data are needed to justify testing and treatment. Our central hypothesis is that a diet moderately high in fructose and salt (FHS) results in hypertension, vascular stiffness and renal dysfunction driven by sympathetic nerve activity (SNA) and/or the renin-angiotensin- system (RAS). We propose to interrogate the mechanism that this neuroexcitation and increased RAS are driven by afferent inputs from the kidney to the brain at the subfornical organ and, thence, to SNA and/or angiotensinergic inputs to heart, vasculature and kidney. Direct interruption of the afferent renal nerves, SNA or pharmacological inhibition of SNA or RAS, alone or in combination, will decrease blood pressure, conduit vascular compliance, and ameliorate cardiac and renal function. We propose three aims: 1) ascertain the respective contribution of afferent vs efferent renal nerves on blood pressure, LV function, vascular compliance, and renal function in FHS rat model, 2) assess the impact of acute or chronic pharmacologic blockade of SNA and/or RAS on LV function, vascular compliance and renal function in FHS rats, and 3) evaluate the contribution of the arterial baroreflex, AT1R and TNFR1 in the subfornical organ that lies outside the blood brain barrier on blood pressure, LV-GLS, PP, PWV, RRI and renal function in rats on FHS diet. RAS and aortic compliance display sexual dimorphism, so we will evaluate Sprague Dawley rats of both sexes. We will use state-of-the-art ultrasonography, real-time renal blood flow and FITC-sinistrin measurements of glomerular filtration rate to assess aortic and renal artery compliance, LV and renal function. We will directly assess the impact of afferent and efferent renal nerves with selective deafferentation vs total denervation in conscious rats. We will validate the in vivo findings by ex vivo myography of aortic rings and assess markers of oxidative stress in vasculature. We will test whether chronic pharmacologic therapies to inhibit sympathetic inputs and/or RAS will also achieve improvements in conduit vascular compliance, cardiac and renal function. Our studies will identify therapeutic interventions that can be translated to screening and treatment of humans with pre-diabetes and stage 1 hypertension to improve vascular and renal function to mitigate MACE and CKD. OMB No. 0925-0001/0002 (Rev. 03/2020 Approved Through 02/28/2023) Page Continuation Format Page

NIH Research Projects · FY 2026 · 2022-03

Abstract In the past 8 years, we have studied a cohort of youth from metropolitan Detroit to investigate the effects of psychosocial stressors and resources on health, with a specific focus on asthma symptoms (Asthma in the Lives Of Families Today, ALOFT study). Our preliminary results from bulk RNA-seq analysis in peripheral leukocytes demonstrate that psychosocial factors are associated with transcriptional changes for a large number of genes, many of them involved in immunological functions. Importantly, we and others have uncovered an important role for blood cell type composition in inter-individual variation in response to psychosocial environments and their effects on immunological health and asthma symptoms. Here, we propose 1) to disentangle the contribution of psychosocial factors and asthmatic state on patterns of transcriptional dysregulation; 2) to investigate the effects of psychosocial factors on transcriptional regulation in blood cell type subpopulations; and 3) to determine the role of genetic variation in modulating these effects and their consequences for asthmatic children's health. To this end, we will use a combination of bulk and single cell RNA-sequencing on immune cells collected from children with asthma and their asymptomatic siblings. The complementary expertise of our team will uncover specific genetic and psychosocial factors associated with increased risk for poor physical health and wellbeing. These results will be important to design personalized medical and behavioral interventions to alleviate disease severity in children with asthma.

NIH Research Projects · FY 2025 · 2021-12

Physical inactivity is a major independent risk factor for cardiovascular disease (CVD) and is now considered the leading cause of premature death (Blair, 2009). Rates of physical inactivity continue to increase along with health care costs to treat CVD. Despite these disturbing trends, the mechanisms by which a sedentary lifestyle leads to CVD are not fully known. CVD is associated with increased sympathetic nervous system activity and overactivity of a brainstem region known as the rostral ventrolateral medulla (RVLM) (Sved et al., 2003;Guyenet, 2006). Sympathoexcitatory responses to direct activation of the RVLM are enhanced in sedentary versus physically active animals (Mischel and Mueller, 2011) and are associated with changes in dendritic branching (Mischel et al., 2014). These data suggest that a sedentary lifestyle may contribute to the development of CVD by increased sensitivity of RVLM neurons. Our long term goal is to understand the central sympathetic mechanisms by which physical inactivity contributes to the development of CVD. This is an important clinical, economic and public health care problem. The overall objective of this application is to define the mechanisms by which physical inactivity increases, and physical activity prevents over-activation of presympathetic neurons in the RVLM. Our central hypothesis is that sedentary and hypertensive conditions each enhance glutamatergic signaling, initiate BDNF-dependent mechanisms and further propagate enhanced glutamatergic signaling; such that in combination, produce clinically relevant increases in sympathetic outflow and blood pressure. This project is expected to shift current paradigms regarding the mechanisms by which physical inactivity and pro-hypertensive stimuli combine to increase sympathetic activity and exaggerate the hypertensive phenotype. We will test our central hypothesis in distinct but interrelated aims using our well- established models of sedentary or active conditions and 2K-1C hypertension with sham-operated rats as controls. Aim 1: Utilize in vivo gene targeting to determine the contribution of BDNF-TrkB signaling in sedentary and 2K1C mediated neuroplasticity in the RVLM. Aim 2: Establish relationships between BDNF and synaptic plasticity-associated mRNA and protein expression in the RVLM of sedentary versus active, normotensive and 2K1C rats using laser capture microdissection of presympathetic RVLM neurons and tract-tracing, triple- immunofluorescent labeling. Aim 3: Quantify glutamatergic tone and neuronal activity in the RVLM of sedentary versus active, normotensive or 2K1C rats using magnetic resonance spectroscopy (MRS) and magnetic resonance imaging (MRI) of the RVLM. Our studies combine state-of-the art techniques with conceptually innovative hypotheses to fill significant knowledge gaps towards understanding two fundamentally important and intertwined, yet unresolved health problems, i.e. physical inactivity and hypertension.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Building upon the strength of existing collaborations and leveraging the intellectual resources and infrastructure across three major research institutions, two in Detroit (Wayne State University and the Henry Ford Health System) and one in Cleveland (Case Western Reserve University/University Hospitals), the ACHIEVE GREATER Center will increase reach into two largely Black communities with an outsized burden of cardiometabolic risk by deploying resources to targeted census tracts; leverage community health workers and pharmacists to help control multiple chronic cardiometabolic diseases; and support the development of early-career stage investigators who are focused on improving chronic cardiovascular disease disparities that are known to exist for Black populations. In addition to conducting a pilot grants program across the three partnering institutions, ACHIEVE GREATER will perform four distinct but closely related special projects that focus on interrupting early stages of pathogenesis in different contexts (e.g., mobile health units versus fixed community locations). Importantly, this work will be nested in a larger epidemiologic study of multi-level cardiometabolic risk factors. Our team will develop a distributed Cloud-based database complete with a customized set of informatics tools that will enable investigators in the heart of each city to robustly profile multi-level risk factors across different domains using both publicly available information and investigator-generated data. Our evidence-based intervention pathways are designed to control risk factors, especially elevated blood pressure, which is the most important modifiable contributor to heart disease - far and away the leading cause of death in our region. By increasing reach in census tracts where uncontrolled cardiometabolic risk factors are most prevalent, our study design optimizes both recruitment opportunities and potential intervention impact. Moreover, the alignment of resources across three institutions will efficiently enhance regional coordination, while increasing the number of research participants and highly trained early-career investigators. If successful, we will have demonstrated a cost-effective and scalable means of reducing health disparities in high risk Black populations through better control multiple chronic diseases.

NIH Research Projects · FY 2025 · 2021-09

Abstract Approximately half of all sexual assaults involve alcohol consumption by the perpetrator, victim, or both. Self- report surveys conducted with victims and perpetrators have provided valuable information about alcohol's role in sexual assault; however, causality cannot be established from correlational designs. When participants are randomly assigned to drink conditions in laboratory studies, causal conclusions can be made regarding the effects of acute alcohol consumption on behavior. The major challenge for experimentalists is to develop proxies for sexual assault that have strong construct validity and experimental realism. Virtual reality environments (VRE) provide the opportunity for participants to become immersed in the simulated environment; thus, participants are expected to behave in ways and to make choices that closely relate to their behavior outside the laboratory. The goal of the proposed research is to build on the promising findings from the PIs recent R21 grant (AA020876) that developed a dating simulation as a new experimental paradigm for examining alcohol's role in acquaintance sexual assault perpetration committed by men against women. The first specific aim of the proposed research involves enhancing the VR dating simulation based on insights from our empirical findings and new technological developments. The updated simulation will be 3-dimensional, with participants wearing head mounted displays that immerse them in the virtual world with their female companion. Changes will be developed and evaluated in focus groups and cognitive interviews with male and female participants to maximize ecological validity. The second specific aim involves systematically evaluating the impact of situational cues manipulated within the virtual reality environment which are expected to evoke the "in the moment" cognitions and feelings that are hypothesized to increase the likelihood of sexual aggression among men predisposed to be sexually aggressive. The third specific aim involves examining the effects of acute alcohol consumption on men's sexually aggressive responses in the virtual reality simulation. Based on the findings from the studies associated with Specific Aim 2, situational factors will be manipulated resulting in a 2 (alcohol condition: sober vs. intoxicated; target BrAC = .08) X 2 (high or low level of cue that affects participants' perceptions of the woman's sexual interest) X 2 (high or low level of cue that affects participants' sense of entitlement and anger after a refusal) design. Risk factors associated with sexual assault perpetration will be assessed in a separate session and are expected to interact with alcohol and cue conditions, such that intoxicated men who are predisposed to sexual aggression (e.g., high pre-existing levels of hostile masculinity) and exposed to sexual interest and entitlement/anger cues are hypothesized to be most likely to be sexually aggressive. Future studies can alter aspects of the simulation to increase generalizability to different populations. The applicants’ long-term goal is to identify modifiable risk and protective factors that can be used to develop evidence-based prevention and treatment interventions to reduce sexual violence.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY A fundamental goal of neuroscience is to determine how different brain regions contribute to behavior (i.e. structure-function relationships). Although significant progress has been made in discovering regional functions that are common across individuals, we know little about how structure-function relationships differ between individuals. Recent evidence suggests that there are many such individual differences in brain activity, with factors like genetics and sex have outsize influence on patterns of brain activity. But the molecular basis of these differences, and their consequences for behavior, are unknown. To begin exploring this new territory, we will (1) identify brain regions whose relationship to behavior is influenced by genetics and sex, and (2) identify neurotransmitters and receptors that underlie the influence of genes and sex on brain function and behavior. We will do this by measuring patterns of whole-brain activity that underlie exploratory behavior and long-term memory recall in adult zebrafish. Zebrafish are a genetically tractable animal model with several characteristics that make them ideal for the present research: a small and simple morphology, a sophisticated behavioral repertoire, and ease of genetic manipulation. As vertebrates, zebrafish retain a high similarity to mammals in genetics and nervous system organization. Given that many mental health disorders manifest as alterations in patterns of neural activity, understanding how individual differences in biology influence brain function and behavior will inform the development of precision approaches for the treatment of disorders localized to the brain.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Neural circuit formation requires a series of highly diverse and specific cell-cell recognition steps, many mediated by cell adhesion molecules (CAMs). Indeed, mutations that disrupt CAMs or their regulation are associated with circuit level neurodevelopmental disorders from dyslexia to schizophrenia. Our model is the mouse retina, an extension of the central nervous system where ~100 types of neurons organize into dedicated circuits that encode the features of the visual world. We focus here on the gamma-protocadherins (γ-Pcdhs), 22 CAMs expressed from a single gene cluster that generate many thousands of distinct homophilic recognition complexes. The γ-Pcdhs are critical regulators of neuronal self-avoidance in starburst amacrine cells (SACs), and cell survival and in many other types of neurons in the retina. The mechanisms through which the γ-Pcdhs serve these functions are unknown, as is the importance of γ-Pcdh isoform diversity. We used a CRISPR/Cas9 approach to generate an unbiased allelic series of mouse mutants with between 1 and 21 intact γ-Pcdh isoforms. From these, we learned that one isoform, γC4, is essential for neuronal survival, suggesting that this isoform functions differently from the other 21. We propose to define the mechanisms of self-avoidance and neuronal survival, and to use our allelic series to determine the level of isoform diversity required for normal neural circuit formation. Our central hypotheses are that: 1) a high level of γ-Pcdh isoform diversity enables neurons to distinguish between “self” and “non-self” to mediate self-avoidance while permitting interaction with neighboring neurons through mechanisms common to all isoforms; and 2) neuronal survival, in contrast, requires interactions specific to the γC4 isoform. In Specific Aim 1, we will use a strategic subset of our reduced-diversity mutants to determine the extent of isoform diversity required for self/non-self discrimination in SACs, neurons essential for the motion detection circuit in the retina. We will analyze this circuit at two levels: A) morphology of contacts between SACs, and B) the electrophysiological function of direction-selective retinal ganglion cells, the downstream neurons in the circuit. In Specific Aim 2, we will define the molecular mechanisms of self-avoidance using in vivo gene delivery to manipulate candidate pathways and map essential domains. In Specific Aim 3 we will uncover the mechanisms through which γC4 promotes neuronal survival. We will use retinal electroporation to map critical protein domains, complemented by a discovery-based proteomics approach to find isoform-specific protein interactions for γC4. These studies will allow us to better understand how the γ-Pcdhs contribute to cell-cell recognition and neural circuit formation in the retina and provide insight into processes disrupted by neurodevelopmental disorders.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT Drug delivery to the brain is restrained by the blood-brain barrier (BBB), a physical and biochemical barrier separating the brain from the circulatory system. Small molecule drugs move across the BBB mainly via transcellular passive diffusion and transporter-mediated active transport. The BBB in brain tumors is disrupted to varying extent, leading to large intra- and inter-individual variability in drug tumor exposure. Mechanistic understanding and prediction of heterogeneous drug penetration across the intact BBB and disrupted blood- brain tumor barrier (BBTB) is of paramount importance to rational drug development and treatment for brain cancer. Given the fact that the rate and extent of drug penetration across the BBB is determined by both biological system characteristics and drug properties, prediction of human BBB/BBTB permeability from preclinical in vitro or animal models is complicated by biological system differences. Hence, the development of innovative approaches is imperative. The in vitro-in vivo extrapolation-physiologically based pharmacokinetic (IVIVE-PBPK) model offers a unique platform that allows simultaneous incorporation of drug- and system- specific parameters into a PK model and enables a priori prediction of individual in vivo kinetic processes based on mechanistic scaling of in vitro data (e.g., in vitro enzyme and transporter kinetics). The overall goal of this project is to develop a mechanism-based PBPK model platform for predicting heterogeneous drug penetration into the human brain and brain tumors. We will employ an integrated translational research approach to achieve this goal, which leverages in vitro pharmacology studies, PK modeling, and clinical trials. Three drugs (AZD1775, ceritinib, and ribociclib) will be used for initial model development and verification, and additional 3 drugs (everolimus, abemaciclib, and LY3214996) will be used for further model validation. These drugs have been or is being evaluated in glioblastoma patients in our clinical trial program. Observed clinical plasma and brain tumor PK data are available for model development and validation. As the first step towards resolving the gap of our knowledge on BBB transporter abundances, which is essential to establishing IVIVE scaling factors for predicting transporter-mediated active clearance at the human BBB and BBTB, Aim 1 is to determine transporter protein abundances in isolated microvessels of non-cancerous cortex as well as contrast-enhancing and non-enhancing glioblastoma specimens. Aim 2 is to determine drug-specific parameters for metabolism, passive transcellular permeability, and interaction with efflux and uptake drug transporters. Aim 3 is to develop and validate a novel 7-compartment permeability-limited brain (7Brain) PBPK model, which accounts for brain and tumor regional physiological differences in blood perfusion, pH, BBB/BBTB integrity, and transporter expression. The 7Brain PBPK model is the first-of-its kind, mechanism-based model platform for the prediction of heterogeneous drug penetration across the human BBB and BBTB. It promises to be a valuable tool to assist the development and design of improved drugs and dosing regimens for more effective treatment of brain cancer.

NIH Research Projects · FY 2025 · 2021-08

Abstract Retinal neurons utilize multiple antagonistic mechanisms to shape visual signal processing, such as ON and OFF signaling, antagonistic center-surround, rod- and cone-signaling, and transient and sustained signaling. Among those signaling mechanisms, ON vs. OFF signaling is coded to morphologically distinct groups of retinal neurons, bipolar, amacrine, and ganglion cells. Retinal neurons that ramify in the inner region of the inner plexiform layer (IPL) respond to the onset of a light stimulus, called ON cells. In contrast, OFF cells ramify in the outer region of the IPL and respond to the offset of a light stimulus. Since being discovered several decades ago, there appeared to be no exceptions to this morpho-physiological rule for ON and OFF signaling. However, recently, some exceptions have been revealed. We found that ON and OFF signaling switches in starburst amacrine cells and bipolar cells. We also found that light adaptation plays a crucial role in the ON and OFF sign switch. In the mesopic light condition, both rods and cones are active, generating a complicated interaction. However, light-evoked responses in retinal interneurons, including bipolar and amacrine cells, have not been systematically investigated over a wide range of ambient light levels. In our proposed studies, we will test the hypothesis that the rod-cone interaction in the mesopic condition converts the ON and OFF signs of light responses in retinal bipolar cells and amacrine cells. The long-term goal of the proposed study is to examine how the ON and OFF sign switch occurs in the interneuron and how this plays a role in shaping spikes in ganglion cells. We will use wholemount retinal preparations to conduct a patch clamp study of bipolar and amacrine cells to determine the visual signal signs in response to various light stimuli. Especially, we will assess the light-evoked excitatory postsynaptic potentials (L-EPSPs) over a wide range of background light conditions. We will also examine the L-EPSPs at the mesopic condition after the tissue is adapted to scotopic or photopic conditions (Aim 1). Also, we will determine how retinal antagonistic signaling systems shape the ON and OFF signs. We will test potential underlying mechanisms, including the rod-cone signaling interaction, the dopaminergic system, the antagonistic surround by inhibitory amacrine cells, and the ON-OFF signaling pathway interaction (Aim 2). Then, we will examine the outcome of the switch by conducting the Calcium imaging from the starburst amacrine cells (Aim 3). Understanding the mesopic vision is essential because both rod and cone dysfunction cause reduced vision in the mesopic vision.

NIH Research Projects · FY 2025 · 2021-07

Project Summary/Abstract The balance between triacylglycerol (TAG) storage and mobilization in adipose tissue and liver is critical to metabolic health as the dysregulation during obesity can produce ectopic accumulation of lipids in muscle and liver, resulting in the progression to diabetes and fatty liver disease (FLD). Thus, an important long-term scientific goal is to understand the tissue-specific molecular mechanisms that control TAG metabolism in order to discover novel therapies. Patatin-Like Phospholipase Domain Containing 3 (PNPLA3) is a protein that is highly expressed in lipogenic tissues, like fat and liver, and is highly upregulated under conditions that promote fat storage. Importantly, a common genetic variant of PNPLA3, I148M, is the greatest known risk factor for developing FLD and its pathological sequelae. In recently published and preliminary data, we demonstrate that α/β hydrolase domain containing protein 5 (ABHD5), an enzyme co-activator, strongly interacts with PNPLA3 and the disease-causing I148M variant is a gain-of-function. Furthermore, the interaction between PNPLA3 I148M and ABHD5 is particularly effective in promoting cellular TAG retention, which likely plays a central role in disease progression. Importantly, the ABHD5/PNPLA3 interaction can be dynamically regulated by endogenous fatty acids and synthetic ABHD5 ligands. This work will examine the basic mechanism of how ABHD5 regulates the function of PNPLA3 and the I148M variant in adipocytes and hepatocytes using high resolution imaging techniques, proximity proteomics and metabolic tracers/lipidomics in conjunction with robust genetic models and an integrative panel of endogenous fatty acid ligands and chemical probes. Our Specific Aims are: 1) To determine the molecular basis for the interaction of ABHD5 with WT PNPLA3 and I148M and the subcellular location and dynamic trafficking of ABHD5 complexes in adipocytes and hepatocytes using high resolution fluorescence and transmission electron microscopy. 2) To determine the location and ligand-dependent protein composition of the ABHD5/PNPLA3 and ABHD5/ PNPLA3 I148M metabolons using nonbiased proximity proteomics and directed immunoprecipitation. 3) To dissect the metabolic function of the ABHD5/PNPLA3 and ABHD5/PNPLA3 I148M metabolons in adipocytes and hepatocytes using gain- and loss- of-function genetics and selective endogenous and synthetic ABHD5 ligands in conjunction with isotope tracers and lipidomics. These goals, which are well aligned with the mission of the NIH will be implemented within a discovery platform that maximizes integration across level of analysis (molecular, organelle, cells and tissues) to provide a robust analysis of ABHD5/PNPLA3 function, thereby improving our understanding of how lipids levels are regulated and identify novel points for therapeutic intervention in obesity related disorders.

NIH Research Projects · FY 2025 · 2021-07

Project Summary Allergies are becoming a major cause of neonatal morbidity, with food allergies showing increased incidences and significantly affecting young infants, some of which can be serious or fatal, and are associated with long- term morbidity, imposing heavy social and economic burdens. For neonatal food allergy, no effective treatment is currently available except avoiding or replacing the offending food, which is often impossible due to the ubiquitous nature of some food components. Hence, there is a critical need to identify effective means of strengthening the immune function of neonates to improve their immediate and long-term health. Human respiratory mucosa and blood harbor secreted immunoglobulin D (IgD). We found that IgD is important in respiratory immune defense by inhibiting mucosal adhesion of pathogens and activating antimicrobial and immune-amplifying functions of basophils. IgD activation of basophils also suppresses IgE-induced allergic functions, and increased food allergen-specific IgD production correlates with protection against food allergy after oral immunotherapy in children. Maternal tetanus, diphtheria, and acellular pertussis (TDaP) vaccine and food exposure in pregnancy induces the production of vaccine- and food-specific IgD that is transferred across the placenta to the fetus in humans and mice. The objectives here are to understand the mechanisms of the placental transfer of IgD and to determine if maternal IgD promotes neonatal immune protection against food allergy. We hypothesize that maternal IgD specific to vaccines or food acts as a specific and prophylactic fetal immune education cue to protect neonates against food allergy. Of note, the basophil-activating and anti- allergic functions are unique to IgD and not possessed by IgG. Employing biochemical and imaging techniques in cell culture, human placenta specimens and mouse models, studies in Aim 1 will mechanistically elucidate the placental transfer of maternal IgD. Aim 2 will determine the function of maternal food-specific IgD in the protection against IgE-mediated neonatal food allergy by integrating neonatal mouse models of IgE-mediated food-induced anaphylaxis with human cord or peripheral blood specimens of newborn babies with or without food allergy in the first year of life. Our study is expected to reveal the unique functions of maternal IgD, an ancient yet still mysterious antibody, in neonatal immune function that maternal IgG does not have, but also have a profound impact on improving neonatal health by directing the design of IgD-targeting maternal vaccines or adoptive immunotherapies.

NIH Research Projects · FY 2025 · 2021-07

Abstract The overall goal of the Fehl laboratory is to develop chemical biology strategies to determine the functional impact of protein modifications during signaling processes. Specifically, cellular metabolism and stress each lead to diverse protein modifications with O-linked N-acetylglucosamine sugar (O-GlcNAc) but no tools are currently able to capture highly dynamic and transient O-GlcNAc events with defined time and spatial resolution. Lack of “time and space” rigor hinders the scientific community from connecting metabolism with disease physiology, including significantly elevated cancer risk in diabetic patients observed for malignancies like breast cancer. In this MIRA application, we pose our strategies to address this critical gap through the development of real-time and space molecular tools that bridge cell metabolism and cancer processes using O-GlcNAc as the keystone. Excellent NIH-funded research has discovered over 2000 O-GlcNAc on proteins in human cells, but current tools rely on disrupted physiology, leading to artifacts, or miss key GlcNAc-driven signaling events that occur before global metabolic rebalancing occurs in less than an hour. We hypothesize that the key drivers of hyperglycemic metabolism and pathology lie within the first few minutes of nutrient and signaling stimulation, which to date is not possible to observe in living cells. Our published work in photochemistry and systems glycobiology support our unique strategies to trigger O-GlcNAc processes in minutes, before O-GlcNAc rebalancing occurs. Our photocaged sugar tool is able to trigger the oncogenic transcription factor NFkB movement between cytosol or endoplasmic reticulum into the nucleus, simulating physiological events that potentially link aberrant insulin and glucose release in diabetes with breast cancer risk. Our real time system can be used to track O-GlcNAc events during insulin signaling for the first time during the rapid, 15-minute pulses of diseased insulin physiology. Another tool for targeted intracellular O-GlcNAc-targeted proximity labeling is able to track O-GlcNAcylated proteins in subcellular space, which no reported tool has the capability to specifically label in live cells. We propose in the next 5 years to develop our “time and space” molecular tools and apply them for unique mechanistic studies in disease biology through NFkB targeting. We actively collaborate with metabolic disease and cancer specialists to ensure disease relevance, as well as with industrial scientists for technology development to expand industrial awareness of O-GlcNAc biology in metabolism-driven disease pathways. The research outputs of this proposal include molecular probes, spatiotemporal strategies, and targets to connect cellular metabolism with signaling. Our enabling chemical strategies have the potential for broad impact in the scientific community by establishing temporal and spatial methods to study protein modifications. Our platforms can be extended to other PTMs and drug targets, such as sialic acid modifications that regulate the interface of cancer metabolism and inflammation. Success will establish a lasting independent research niche.

NIH Research Projects · FY 2025 · 2021-07

Research on human health has fundamentally changed in the last 20 years. Modern genomics and proteomics technologies have carefully characterized health-related events, which has opened the door to asking more complex questions about normal and disease states. Correspondingly, to confront complex research challenges, the career opportunities to Ph.D. scientists include multiple non-academic positions (e.g., industry, biotechnology, government, publishing). Along with this research progress and changing career landscape, the biomedical culture has placed greater emphasis on quantitative data analysis, rigor and reproducibility, and transparency to ensure the quality and accuracy of research findings and safety to minimize risk to scientists and the environment. To take advantage of this unprecedented opportunity to thoroughly dissect biological systems with high standards of rigor and many career perspectives, a biomedical work force capable of multi-disciplinary, team-driven, and problem-focused thinking is needed. In total, this modern world of biomedical research requires a rethinking of the Ph.D. training experience. To address the changing needs in biomedical work force training, we have created a Chemistry Biology Interface (CBI) training program at Wayne State University (WSU). WSU has the ideal campus environment for a scientifically rigorous training experience. CBI faculty among six departments and three colleges offer a breadth of research expertise for graduate training. WSU has in place a series of professional development activities to augment career preparation. Building upon these strengths, the CBI program will augment the graduate training within core disciplines by promoting multi-disciplinary research knowledge and collaboration facilitated by cross-campus mentoring and peer-to-peer advising. These outcomes will be achieved through a series of CBI activities that center around an individualized “My Ph.D.” theme. The foundational activity is an annual "My Ph.D." workshop focused on career skills surveying, assessment, and planning, which will be led by CBI student peers and faculty for multi-level mentoring. Complementing the individualized skills evaluation, multi-disciplinary research, scientific rigor, communication skills, and peer cohort building/networking will be accomplished through annual CBI symposia, biweekly seminar activities, a team- taught and literature-based CBI course, a co-mentoring Ph.D. committee structure, early and sustained academic advising, internal and external research experiences (ATTRACT), and community outreach events (ACT OUT). Through these meaningful activities, WSU Ph.D. graduates will form a strong scientific identity with discipline-specific skills and informed multi-disciplinary perspectives. The combination of research breadth, skills depth, and enhanced mentoring facilitates the confident, solution-oriented, and collaborative thinking needed to address complex biological questions and migrate through a multifaceted career backdrop.

NIH Research Projects · FY 2025 · 2021-07

Abstract Although many studies have demonstrated correlations between cytoskeletal dynamics, genome organization, and gene expression, the underlying mechanisms linking them remain unclear. Recently, we discovered crosstalk between the Wiskott-Aldrich syndrome protein family verprolin homolog (WAVE) regulatory complex (WRC), which promotes actin polymerization, and the Spt-Ada-Gcn5-acetyltransferase (SAGA) complex, a transcriptional coactivator. Their relationship is established through the sharing of subunits comprising the SAGA deubiquitinase module, including the deubiquitinase Non-stop. Deubiquitinase module mutations—for example, polyglutamine expansion in ataxin 7 (ATXN7)—lead to a spectrum of phenotypes not explained by SAGA’s transcriptional coactivator function, including nervous system degeneration and blindness. In both the nervous system and the eye, the WRC is an essential promoter of actin polymerization, which is facilitated by the constitutively active enzymatic subunit WAVE. WAVE activity and localization are regulated by the remaining WRC subunits, ensuring spatial and temporal control of actin polymerization. Misregulation of actin polymerizing complexes results in a similar spectrum of phenotypes as seen in SAGA mutants, including nervous system degeneration and blindness. This suggests that SAGA is important in the nervous system and eye because it is required to control the WRC in these tissues. We found that the SAGA deubiquitinase module leaves SAGA to bind the WRC. There, Non-stop deubiquitinates WAVE, increasing its level in both the cytoplasm and the nucleus. Therefore, we hypothesize that SAGA controls WRC complex composition, amount, and location; and it is through these activities that SAGA accomplishes functions we had previously attributed to SAGA alone. This hypotheses will be investigated in three aims. First, we will identify and characterize nuclear WAVE-containing complexes through affinity purification and column chromatography coupled to mass spectrometry. Because WAVE activity is regulated in part by its localization, the locations of these complexes will be determined, and the effects of Atxn7 polyglutamine expansion on complex composition and localization will be tested. Second, interactions between the SAGA deubiquitinase module and WRC will be disrupted in cells and flies to determine which SAGA/WRC functions require them. Lastly, the effects of SAGA deubiquitinase-WRC interactions on blindness and neurodegeneration will be investigated in flies, by disrupting them and measuring phenotypes characteristic of Atxn7 polyglutamine expansion. These studies will provide novel insight on the causes of neurodegeneration and blindness, in addition to the links between transcriptional and cytoskeletal regulatory complexes.

NIH Research Projects · FY 2025 · 2021-05

It has been widely accepted that distinct epithelial to mesenchymal transition (EMT) phenotype and cancer stem cell (CSC) properties as well as the immunosuppressive tumor microenvironment (TME) in triple negative breast cancer (TNBC) subtype account for the aggressive behavior of this disease. Although increased levels of tumor- infiltrating lymphocytes (TILs) in TNBC predicted better clinical outcome, the majority of these patients display progressive disease due to the immunosuppressive TME. Although the clinical relevance of TME/pre-metastatic niche in disease progression has been well recognized, the molecular mechanisms that regulate these processes remain elusive. Preclinical and clinical data provide compelling evidence that immune cells of myeloid origin (macrophages, neutrophils, MDSCs) are major components of the TME and predictive of poor prognosis as well as therapeutic resistance. Therefore, further research is required to understand the underlying molecular mechanism of formation of immunosuppressive TME/pre-metastatic niche and its role in disease progression and therapeutic resistance. Our primary objectives in this application are; to determine how stress-induced HSP70 regulates two fundamental processes; i) protecting tumor cells from cytotoxic cell death by inducing an epithelial mesenchymal transition (EMT) and cancer stem cell (CSC) phenotype and ii) generating a permissive microenvironment via the modulation of immunosuppressive myeloid cells. Our central hypothesis is that A20 induced HSP70 in TNBCs protects tumor cells from cytotoxic cell death while inducing an EMT phenotype and inflammatory cytokines which in turn promote the accumulation of immunosuppressive MDSCs. Therefore, targeting HSP70 will have a dual activity on tumors and MDSCs. Our rationale is that the identification of molecular mechanism(s) that sensitize tumor cells to cytotoxic agents while reversing immunosuppression will improve the effectiveness of currently available therapeutics. We previously demonstrated that growth of tumors at metastatic sites is dependent of granulocytic MDSCs and suppression of anti-tumor responses and thus blocking HSP70 in combination with standard of care and/or checkpoint inhibitors could have significant clinical benefit. Based on these concepts we propose to test our hypothesis by investigating the following specific aims: Aim 1 will test the hypothesis that a reciprocal A20/HSP70 signaling axis provides cytoprotection to tumor cells by inducing EMT/CSC phenotype in TNBC subtype. Aim 2 will test the hypothesis that HSP70 regulates immunosuppressive MDSC induction and acitivity. Aim 3 will determine the molecular mechanism and functional importance of HSP70 in generation of TME and pre-metastatic niche. At the completion of our proposal, we expect to elucidate the molecular mechanism by which HSP70 cytoprotects tumor cells from cytotoxic agents by inducing EMT/CSC phenotype while regulating the immunosuppressive MDSCs in response tumor secreted cytokines facilitating the tumor progression. It will also determine whether blocking HSP70 potentiates the efficacy of the chemotherapies and/or immunotherapy in syngeneic mouse models representing TNBC subtype.

NIH Research Projects · FY 2025 · 2021-04

Despite the steady decline in cardiovascular diseases (CVD) morbidity and mortality in the US in the last few decades, African American (AA) adults bear a disproportionate share of cardiovascular disease (CVD) burden. Psychosocial factors and emotional reactivity to them are believed to contribute to the etiology and progression of CVD through their effects on health behaviors, the stress-responsive neuroendocrine axes, and immune processes. These factors are particularly salient for urban-dwelling middle-aged and older AAs, who experience unique stressors and are more likely to live in situations of socioeconomic disadvantage than Whites. However, psychosocial factors and their link to CVD risk, and inflammation more broadly, have been remarkably understudied among AA adults. A fine-grained characterization of the daily stressors, health behaviors, and emotional responses related to CVD—and understanding of the situational contexts in which those occur—will significantly advance the science of CVD risk. Accordingly, the purpose of the proposed project is to identify and conceptualize—through a mixed-method approach—the psychosocial stressors most salient for this population and to model the daily psychological, behavioral, and biological pathways through which these factors may exacerbate CVD risk among middle-aged and older AAs. By adopting a prospective (two waves over two years) and multiple-time-scale design (daily assessments nested within waves), we will test this idea in a sample of 500 asymptomatic AAs aged 55-75 years living in Detroit. We will also use semi-structured interviews to collect qualitative data from 60 participants to contextualize the quantitative results. Our central hypothesis is that interpersonal stressors will predict decreases in resting heart rate variability and increases in resting blood pressure, poor sleep, chronic physiological stress (hair cortisol), and inflammation (basal and stimulated cytokines and basal CRP) by altering daily affect, daily health behaviors, and daily physiological stress (salivary cortisol). We propose to increase the innovation of our work by (1) using a smartphone-based ecological momentary assessment protocol to measure psychosocial stress, (2) including a sequential explanatory mixed-method design, (3) adopting a multiple-time-scale research design and a standardized measure of neighborhood stress created ad hoc for Detroit, and (4) simultaneously considering multiple measures of physiological stress, inflammation, and surrogate endpoints of CVD. The rationale for the proposed research is that once a clear picture of the daily psychosocial risk factors for CVD is formulated, and their biological intermediaries are identified, more culturally and individually tailored treatments can be developed to reduce CVD in this population.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY/ABSTRACT Research Project: Excess body weight is a contributing factor in up to 20% of all cancer deaths in women. In particular, endometrial cancer (EC) incidence has risen steadily over the past two decades concomitant with the rise in global obesity rates. EC is a multifactorial disease, as both obesity and somatic driver mutations play causal roles. Mutations in the phosphoinositide 3-kinase (PI3K) pathway are present in over 80% of all EC. However, obesity or PI3K pathway mutation alone are insufficient for EC pathogenesis. A major gap in knowledge is how genetic mutations alter EC risk in obese women, and evidence points towards impaired protein quality control as an important causal mechanism. Obesity causes systemic endoplasmic reticulum stress (ERS), which is triggered by the accumulation of unfolded proteins. If proteostasis cannot be achieved, the unfolded protein response induces cell death. Due to increased aromatase activity, obesity creates an “unopposed estrogen” phenotype, which drives EC and results in ERS in the uterus. The PI3K pathway antagonizes ERS and may promote cell survival under these conditions. Here, I propose to study the casual mechanistic relationship between obesity and PI3K pathway mutations in EC pathogenesis. My central hypothesis is that obesity alone causes ERS in the normal uterus, resulting in cell death, but when a PI3K pathway mutation occurs in the endometrium of obese women this stress response does not happen, resulting in uncontrolled cell growth and cancer. In this application, I propose to 1) Determine the relationship between PI3K mutation and ERS in the endometrium 2) Characterize the role of estrogen-induced ERS in EC 3) Determine the role for metabolism-induced ERS in EC prevention. Career Goals: My career will be focused on cancer research. In graduate school, I developed novel cancer therapeutics targeting nucleotide biosynthesis under the guidance of Dr. Larry Matherly. As an American Cancer Society Postdoctoral Fellow in Dr. Ronald Chandler’s lab, I uncovered the mechanism by which common somatic mutations in the endometrial epithelium result in myometrial invasion in EC. My goal is to secure a tenure-track faculty position at a leading cancer research institution, and to develop a research program exploring the relationship between obesity and mutation in EC pathogenesis to identify targets for active preventative intervention. Career Development and Environment: The K99/R00 award will secure the necessary time and training to develop my career as a successful independent investigator. My mentors will be Drs. Ronald Chandler, Jose Teixeira and Victoria Bae- Jump, experts in the field of gynecologic oncology. Activities planned will focus on gaining new expertise in obesity research. Michigan State University and the Van Andel Research Institute are the ideal location for this training, being home to many experts in gynecologic oncology, cancer metabolism, mouse modeling and bioinformatics. The Michigan Nutrition Obesity Research Center will provide additional education and training in the fields of metabolism and obesity research.

NIH Research Projects · FY 2025 · 2021-02

The mission of Wayne State University (WSU), to create and advance knowledge, prepare a diverse student body to thrive, and positively impact local and global communities, is very much in line with the goals and vision for the Maximizing Student Development (IMSD) program. Located in Detroit, Michigan, WSU has historically been a stabilizing institution in the City, and is now at the forefront of the revitalization taking place in the region. WSU is committed to driving this growth, and over the last 5 years has recruited over 50 new faculty members in the sciences alone. New faculty are drawn to WSU for various reasons, including the diversity of the region, the exceptional facilities, and the success of our faculty, as evidenced in part by a 30% increase in federal funding since 2014. WSU is well-positioned to request continued support for its successful IMSD program, now transitioning from a R25 to a T32, at the level of 10 graduate students. The goal of the WSU program is to provide targeted training and mentoring that is individualized to meet the needs of a diverse body of graduate students to facilitate successful completion of their Ph.D. degrees. Ultimately, these IMSD students will develop careers in the biomedical sciences, and will go on to serve as mentors to future generations. Trainees will be selected in their first or second year of graduate study and supported for up to two years. The IMSD T32 program will continue to focus on developing and enhancing academic skills and professional career development. IMSD graduate students are integrated into our learning community, and remain active throughout their entire training at WSU, with enhanced opportunities for teaching and mentoring. Specifically, we aim: 1) To provide our graduate students with a structured community to facilitate and enhance professional bonds that will enable each student to learn, develop, and incorporate the skills necessary for a successful future in biomedical careers; 2): To provide a program to enhance individual academic program success coupled with mentored research; and 3): To provide opportunities for peer-to-peer mentoring, near- peer mentoring, and undergraduate teaching experiences in our learning community. These activities are intended to provide opportunities for IMSD student trainees to develop skills in presentation, teaching, and mentoring that will not only maintain their focus on career goals, but also develop professional skills that they will utilize in their future research/academic careers. We propose to build on the successes of our long- established IMSD program with modifications based on an evolving demographic and social culture of our students. This IMSD program will provide academic development and research experience at a large, urban, research institution with a very diverse student population to a more personal level for our trainees.