Temple Univ Of The Commonwealth

NIH Research Projects · FY 2026 · 2023-08

Polydrug abuse (especially alcohol use disorder, AUD, and smoking) are known individually to compromise the lung alveolar-endothelial barrier (AEB) and the blood brain barrier (BBB). Very limited knowledge exists regarding damage in lung and brain due to electronic cigarettes (e-Cig) in combination with AUD. E-Cig have become popular, yet very limited data indicate that they cause endothelial dysfunction and result in a pro-inflammatory phenotype in macrophages and endothelium in lungs. While e-Cig are known to be addictive, their effects on the brain and cognition are essentially unknown. Our data show that chronic e-Cig exposure in mice enhanced permeability of the BBB and neuroinflammation, diminished expression of a key glucose transporter and tight junction protein on brain endothelium, and impaired cognition. Preliminary data indicate that the combination of alcohol/e-Cig exposure in vivo caused enhanced AEB permeability and amplified neuroinflammation/BBB compromise. We found that e-Cig and alcohol impair AEB and BBB via the same mechanism including mitochondrial dysfunction, Ca2+ accumulation, and ATP extracellular release, potentially mediated by purinergic receptor, P2X7, in cellular components of AEB/BBB. Using innovative in vitro 3D systems of AEB and BBB and relevant animal models, we test the hypothesis that BBB and AEB injury in e-Cig/alcohol exposure are mediated through the P2X7 receptor. In aim 1, we will screen the magnitude of injury (mitochondrial dysfunction, Ca2+ increase and ATP release) by various types of e-Cig in combination with alcohol on human brain and lung endothelial and lung epithelial cells. Then, we will define mechanisms of demise using innovative 3D in vitro constructs of lung and brain microvasculature, functional assays, assessment of mitochondrial functions and expression of key molecules supporting BBB and AEB. We will investigate the contribution of activation of the purinergic P2X7 receptor in e-Cig/alcohol induced BBB/AEB dysfunction. The 2nd aim will study in vivo lung injury after chronic alcohol feeding and e-Cig vaping evaluating AEB permeability, expression of barrier supporting molecules, inflammatory responses (immunohistochemistry, protein/mRNA, bronchoalveolar lavage). P2X7 knockout (KO) animals will allow dissection of the role of this receptor in pulmonary dysfunction. The 3rd aim will decipher combined in vivo effects of BBB function, expression of barrier-mediating molecules, and neuroinflammation. The same experiments performed in P2X7 KO mice will determine the importance of this pathway in CNS injury. Markers of lung injury and BBB damage will be measured in blood and correlated with signs of end-organ pathology.

NIH Research Projects · FY 2026 · 2023-07

PROJECT SUMMARY/ABSTRACT Latino adolescents experience higher rates of behavioral health risks compared to their non-Latino Black and White peers, particularly for substance use and suicidality. Poor mental health and substance use have been attributed to experiences of childhood adversity and a range of multilevel stressors. Advances in the field of youth mental health research has been largely focused on individual and interpersonal factors. Few studies examine the impact of social and environmental contexts, such as those related to educational access, on youth mental health. Population-level interventions, and the educational access and economic trends that shape those contexts, also affect youth stress and access to resources, increasing mental health risk for adolescents. Some of these factors shape the social environment, amplifying experiences of stress and mental health issues. State-level contexts that increase access to health, social and educational resources are key factors in shaping the mental health of Latino youth. Research examining pathways between population-level interventions and the mental health of youth is needed. The proposed study will fill gaps by applying a transdisciplinary approach to understand and intervene upon an important health disparity issue. The objectives of this research are to: (1) construct a state-level dataset focused on population-level educational access interventions for students; (2) use quasi-experimental methods to quantitatively estimate the effects of population-level interventions, economic indicators and educational trends on youth mental health and qualitatively explore how population-level interventions shape the social lives, educational trajectories and mental health of all youth in the US; and (3) develop community-informed recommendations for prevention and intervention solutions. With a focus on interdisciplinary collaboration, Temple University offers early career scholars the intellectual and practical resources to develop into successful independent investigators. The overarching training plan for this K01 application serves to consider the effects of population-level behavioral interventions on the mental health of youth in the US. The training plan is designed to develop the candidate’s skills in population-level interventions, quasi-experimental design methods and community-engaged research, to develop ground-level recommendations for prevention and intervention. By engaging in coursework, trainings, applied experiences, and national conferences, complemented by a transdisciplinary team of mentors, Dr. Villamil Grest’s training and career development plan provides an opportunity to advance her interdisciplinary skills to understand the upstream, population-level factors that affect the health and well-being of youth. To achieve success as an independent scholar and researcher, this K01 will provide skills for the PI to become a multimethod investigator, examining the social and population-level interventions that contribute to youth mental health disparities, and to inform population-level and community-engaged prevention.

NIH Research Projects · FY 2022 · 2023-07

PROJECT SUMMARY Alpha1-antitrypsin deficiency (AATD) is the most common genetic cause of liver disease in children and the most frequent inherited indication for liver failure and transplantation in the pediatric population. However, the clinical course of AATD-related liver disease is highly variable. The majority of infants with homozygous severe AATD (PiZZ) clinically recover in early childhood through the unknown adaptive mechanisms. Epidemiological studies give rise to three outstanding questions in the field. They are: (1) What are the mechanisms of adaptation to the misfolded AAT accumulation in the lumen of ER? (2) Do the mechanisms of adaptation only response to misfolded AAT or also target other unfolded proteins? and (3) Can we design therapeutic strategies to directly use these mechanisms of adaptation? Here, we identified a novel alpha1-antitrypsin deficiency adaptive mechanism, ER proteotoxic stress-m6A pathway or ERm6A : Unfolded Alpha1-antitrypsin protein accumulation induces N6-adenosine- methyltransferase 14 (METTL14) elevation to increase m6A mRNA methylation of C/EBP Homologous Protein (CHOP), which suppresses CHOP translation and reduces expression of its downstream pro-apoptotic target genes, apoptosis and liver injury. We propose that ERm6A regulates ER proteotoxic stress at the epitranscriptional level, through a mechanism that is distinct from any other previously described ER proteotoxic stress-associated signaling pathway, including the well-known canonical (transcriptional and translational level) UPR pathway. We will test the central hypothesis that METTL14, induced by unfolded protein accumulation, suppresses cell death and liver hepatotoxicity (Aim 1.1) by modulating CHOP 5′ UTR m6A modification (Aim 1.2). This proposal addresses the mission of the NICHD by rigorously investigating the molecular mechanisms of AATD-induced liver injury with the potential to improve health for large numbers of children.

NIH Research Projects · FY 2026 · 2023-07

PROJECT SUMMARY The worldwide epidemic of nonalcoholic fatty liver disease (NAFLD) has become a severe and costly threat to public health. The current therapeutic options for NAFLD are exceedingly limited, especially for its severe form - nonalcoholic steatohepatitis (NASH), which has no FDA-approved therapy. There is an urgent need to identify novel therapeutic targets for NASH treatment. Thioredoxin interacting protein (Txnip) is a stress-induced gene, and it has been implicated in NASH. However, the role of Txnip in NASH is controversial, which may be attributed to the following two issues. Firstly, there are two genes in the Txnip gene locus: Txnip and Txnip antisense long non-coding RNA Gm15441. Genetic deletion of Txnip also deletes part of Gm15441. Secondly, Txnip plays critical roles in other metabolic organs, which interferes with its function in the liver. Therefore, previous studies using Txnip global knockout mice were not able to reveal the liver-specific role of Txnip in NASH. Our preliminary data demonstrate that Txnip protein is abnormally accumulated in NASH mouse livers. Our preliminary studies also suggest that Txnip may promote hepatocyte apoptosis. Given that excessive hepatocyte apoptosis drives NASH development, therefore, we propose a central hypothesis that inhibition of hepatic Txnip alleviates NASH. In Aim 1, an antisense RNA based Txnip translational inhibitor will be developed for NASH treatment. In Aim 2, we will dissect the functional role and mechanism of Txnip in NASH mouse liver. In Aim 3, we will investigate a novel mechanism that causes Txnip protein accumulation in NASH mouse liver. Completion of this proposal will provide critical insights into the functions and mechanisms of Txnip in NASH development. These studies will also lead to the development of novel therapeutic strategies for NASH treatment.

NIH Research Projects · FY 2025 · 2023-07

PROJECT SUMMARY Cardiovascular disease (CVD) remains the leading cause of death, and there is a critical need to determine underlying mechanisms and develop novel therapies for treatment. Small Cajal body-specific RNAs (scaRNAs) are evolutionarily conserved non-coding RNAs that guide biochemical modification on specific nucleotides such as pseudouridylation (Ψ) and 2′-O-ribose methylation (2′-OMe). These modifications are essential in the post- transcriptional changes and functions of spliceosomal small nuclear RNAs (snRNA). Recent studies highlight a crucial role of scaRNA in preserving spliceosome fidelity and cardiac development; however, the role of scaRNAs in acute cardiac injury and its repair has not been established. This proposal aims to determine scaRNA’s role in cardiovascular processes in a mouse myocardial infarction (MI) model that could develop new tools for treating MI. Our central hypothesis is that overexpression of scaRNA18 in the post-MI heart promotes positive cardiac remodeling by inducing therapeutic angiogenesis via 2′-OMe of U4 snRNA and regulating Wilms tumor1 (WT1) expression, thereby conferring cardioprotection and improving cardiac function. Our preliminary data demonstrates 1) decreased scaRNA18 expression in post-MI mouse hearts using unbiased small RNA sequencing, 2) scaRNA18 expression exclusively downregulated in isolated cardiac endothelial cells (EC) in post-MI mouse hearts, 3) scaRNA18 knockdown (KD) impairs tube formation and induces apoptosis in mouse cardiac ECs without induced stress, 4) scaRNA18 overexpression enhances EC angiogenesis and inhibits stress-induced EC apoptosis in vitro, 5) scaRNA18-overexpressing cardiac EC extracellular vesicles protect iPSC-cardiomyocyte cell death via angiogenic proteins, 6) 2’-O-methylation levels were reduced in MI hearts and in cardiac ECs from scaRNA18 KD mice, 7) mutant scaRNA18 lacking binding sites to guide 2′-OMe on U4 snRNA induces EC cell apoptosis without induced stress, 8) unbiased mass spectrometry results revealed that scaRNA18 reduces WT1 expression levels in scaRNA18 KD cardiac ECs, 9) WT-1 KD in the presence of scaRNA18 overexpression nullified scaRNA18 mediated EC survival effects, and 10) overexpression of scaRNA18 by VE-cadherin-driven AAV9 reduces infarct size after ex vivo cardiac ischemia/reperfusion injury and improves cardiac function post-MI mice (in vivo). The hypothesis that scaRNA18 is a critical component in cardiac function and repair will be tested in the following three specific aims: in Aim 1, we will define the role of scaRNA18 in cardiac EC function and cardiomyocyte survival; in Aim 2, we will determine the molecular mechanisms by which scaRNA18 regulates cardiac EC function and cardiomyocyte survival, and in Aim 3 we will use post-MI mice to demonstrate the critical role of scaRNA18 in myocardial injury repair. The proposed studies could establish scaRNAs as novel therapeutic targets for MI.

NIH Research Projects · FY 2025 · 2023-07

PROJECT ABSTRACT 6.2M Americans over the age of 65 suffer from Alzheimer’s disease (AD) and this number expected to double in ~30 years. AD is characterized by aphasia, loss of fine and gross motor function, and rapid cognitive decline. The widely favored “amyloid hypothesis” of AD posits that accumulation of fibrillar amyloid beta (Aβ) plaques in the brain parenchyma drives AD pathogenesis. However, the amyloid pathway has proven to be an ineffective therapeutic target in numerous clinical trials and AD remains clinically intractable, highlighting the urgent need for deeper understanding of the underlying mechanisms of disease. Our lab previously reported that mitochondrial calcium (mCa2+) overload promotes AD pathogenesis. mCa2+ homeostasis is maintained through regulation of mCa2+ uptake through the mitochondrial calcium uniporter channel (mtCU) and mCa2+ efflux through the mitochondrial Na+/Ca2+ exchanger (NCLX). Human cortex from sporadic AD patients demonstrates >70% reduction in NCLX expression. Genetic rescue of mCa2+ efflux via hippocampal neuron-specific expression of NCLX protects against mCa2+ overload, ROS-stress, Aβ and tau deposition, and cognitive decline in AD mutant mice. We interpret remodeling of mCa2+ transport as a compensatory response to an early pathologic stress (e.g., energetic crisis, aging, genetic predisposition) to increase ATP bioavailability. Over time, this response turns maladaptive and promotes pathologic mCa2+ overload. mCa2+ overload causes excessive production of reactive oxygen species (ROS), metabolic derangement, and cell death, all hallmarks of AD. Although a robust connection between neuronal mCa2+ overload and AD pathogenesis has been established, how altered regulation of mCa2+ uptake promotes or protects against AD pathology remains completely unexplored. Our preliminary data demonstrates MICU3 expression is significantly reduced by ~50% in multiple cortical regions of samples isolated from sporadic AD patients. Further, MICU3 expression is reduced >90% in the cortex of 1 year-old. 3xTg-AD mutant mice. This proposal hypothesizes that loss of neuronal Micu3 contributes to aberrant mtCU-mediated mCa2+ uptake, resulting in mCa2+ overload, metabolic derangement, neuronal dysfunction, and cognitive decline in AD. To address this hypothesis we will utilize newly generated neuron-specific MICU3 knockout mouse lines to measure if knockout of MICU3 alone is sufficient to cause neurodegeneration. Subsequently, we will use our newly developed cre-inducible MICU3 overexpression mouse line to see if rescuing MICU3 levels shortly after onset of cognitive decline in the APPNL-G-F mouse model of AD is sufficient to mitigate or reverse AD pathology. These studies will be followed up with a series of mechanistic in vitro studies to determine the molecular mechanism of MICU3-mediated neuronal dysfunction in AD. The role of MICU3 in physiology and disease states, including AD, is unknown; coupled with our findings that altered mCa2+ handling is a pathologic feature of and promising therapeutic target for AD provides strong rationale for this proposal.

NIH Research Projects · FY 2025 · 2023-06

Live donor kidney transplant (LDKT) is the preferred treatment modality for patients with chronic and end-stage kidney disease. LDKT is less expensive than prolonged dialysis and offers improved mortality and morbidity over either dialysis or deceased donor kidney transplants (DDKT). However, LDKT continues to be underutilized, despite efforts to increase access and interest. Decades of research point to patient-provider communication as a contributing factor to observed disparities in health and healthcare outcomes. To date, however, no attempt has been made to gauge the impact of the communication occurring during transplant evaluation consultations on LDKT outcomes. The long-term goal of the proposed study entitled, Improving Utilization of Live Donor Kidney Transplant through Effective Patient-Provider Communication, is to increase knowledge of and interest in LDKT for patients in need of kidney transplants. We propose a community-engaged, mixed-methods study employing a concurrent triangulation design to identify the specific communicative behaviors that result in live donor inquiries and evaluations, and actual LDKTs, providing critical information to the design of an intervention to improve patient-provider communication about LDKT. Specifically, we will simultaneously quantitatively assess patient and provider factors with established and hypothesized associations with receipt of LDKTs, and qualitatively assess discrete elements of patient-provider communication occurring during transplant evaluation consultations (Aim 1). Brief quantitative surveys administered before and after medical consultations held as part of the evaluation for transplant candidacy will capture providers’ (N=52) confidence and comfort discussing LDKT and patients’ satisfaction with the consultation, medical mistrust, health literacy, and LDKT knowledge, attitudes and readiness. We will also audiorecord transplant evaluation consultations for 120 patients across the two study sites (60 per site) – Saint Barnabas Medical Center (NJ) and Temple University Hospital (PA), and qualitatively assess the communication occurring during the consultations. We will use the findings to inform development of the content and format of a communication skills training for transplant providers and evaluate the direct and indirect effects of the training on patient-reported and LDKT process outcomes (Aims 2 & 3). Intervening at the provider level is both practical, given that all transplant candidates already must undergo this consultation, and efficient, given that a single transplant physician can evaluate >100 transplant candidates per year. Thus, the results of this innovative study have the potential to increase access to LDKT for Black patients currently awaiting kidney transplant. Improving communication during the transplant consultation may prove to be an effective and efficient means of increasing utilization of LDKT.

NIH Research Projects · FY 2026 · 2023-06

SUMMARY Atopic dermatitis (AD) is an inflammatory skin condition that paradoxically increases the risk of skin infections. The most common bacterial infection of clinical significance is Staphylococcus aureus, while the herpes viruses herpes simplex virus (HSV) and varicella zoster virus (VZV) greatly affect quality of life and can lead to severe infections requiring hospitalization. While it is known that AD dysregulates the barrier function of the skin, how this impacts immunity against microbial pathogens is poorly understood. Improved insight into how immune mechanisms in the skin are initiated and resolved may underpin future development of preventive and therapeutic strategies. Interleukin-36 (IL-36) represents IL-36a, IL-36b and IL-36g, which are related to IL-1a and IL-1b. Elevated IL- 36 levels are associated with S. aureus colonization and severe AD. Using HSV1 as a model herpes virus, we published studies on the roles IL-1 and IL-36 play in initiating protective immune responses. Additional data from our lab identified critical activities that contribute to inflammation and the elimination of S. aureus. These findings improved our understanding of functions that are common for both IL-36 and IL-1. New unpublished data indicate mechanisms whereby the two cytokine groups also have distinct functions and how they may contribute to AD when dysregulated. To elucidate these mechanisms, we will here use mouse models of in vivo skin infections with HSV1 and S. aureus to identify the IL-1 and IL-36 sensing cell types essential for immune responses that promote inflammation and restrict the pathogens. These studies will involve a new floxed IL-36 receptor mouse strain we developed, in conjunction with an existing floxed IL-1 receptor strain. Complementary and mechanistic studies will also be conducted in human and mouse primary cells. A comprehensive in-depth understanding of these systems is needed to successfully suppress immune responses therapeutically, while at the same time maintaining functional protective immunity against pathogens such as HSV and S. aureus.

NIH Research Projects · FY 2024 · 2023-06

PROJECT SUMMARY Making healthy food choices and eating in moderation during early childhood are central to obesity prevention and are thought to require effortful and goal-directed self-regulation. Appetite self-regulation (ASR) has been described as involving children’s use of eating-specific, “top-down” cognitive processes to moderate “bottom- up” biological drives to eat. Much of the research to date on ASR has focused on the role of bottom-up drives in shaping children’s behavioral susceptibility to obesity. Alternatively, little is known about the cognitive- developmental processes that shape children’s ability to make healthy food choices and eat in moderation during early childhood. Current perspectives hold that ASR is distinct from general self-regulation (e.g., executive functioning [EF]) among children, highlighting the large gap in scientific understanding of cognitive developmental influences on healthy eating and obesity prevention during the preschool years. The goal of this R21 exploratory investigation is to produce rigorous evidence of cognitive developmental influences on healthy eating behaviors (i.e., healthy food choices, eating in moderation) and weight status during preschool through the development of new measures of top-down ASR. Preschool is an important period for studying top-down ASR given the significant socialization of eating behaviors and rapid maturation of top-down regulatory processes that occur during this period. Participants will be 150 preschoolers (75 with normal weight, 75 with overweight or obesity) and their primary caregiver. Given well- documented socioeconomic disparities in self-regulation, diet quality, obesity among children, we will oversample families with low-income backgrounds. We will adapt existing measures of inhibitory, working memory, and attention shifting–core aspects of EF – to develop new measures of eating-specific, top-down ASR. ASR/EF associations with laboratory-based observations of children’s eating behaviors, body mass index z-scores, and questionnaire-based measures of food parenting will be assessed. Aim 1 will adapt well- established objective observational measures of top-down EF to assess top-down ASR regulation among preschoolers. Aim 2 will examine the protective role of top-down ASR in making healthful food choices, eating in moderation, and weight status among preschoolers. Given that parenting represents a socialization pathway that can hinder or facilitate self-regulation in children, Aim 3 will evaluate associations of food parenting structure and autonomy support with top-down ASR among children. Raising children to make healthy food choices and eat in moderation in the current obesogenic environment may require more explicit involvement of cognitive-developmental processes around eating than has been previously appreciated. The findings of this investigation will yield novel scientific directions for obesity prevention by elucidating cognitive developmental influences on healthy food choices, eating in moderation, and weight status among preschoolers.

NIH Research Projects · FY 2026 · 2023-06

This proposal is to delineate the novel function of apical polarity complex proteins in cortical progenitor mitosis and uncover a new pathogenic mechanism of microcephaly caused by genetic mutation of Pals1 (protein associated with Lin7 1, also known as MPP5). PALS1, which has been incriminated only recently as a gene responsible for microcephaly in humans, encodes a component of the evolutionarily conserved apical polarity complex. Our multiple Pals1 genetic models using different Cre drivers consistently demonstrate overwhelming cortical cell loss because of compromised cell viability. However, the cellular and molecular defects behind the massive cell death found in Pals1 mutants remain unknown. Through extensive time-lapse imaging, we found that Pals1 loss causes abnormally lengthened mitotic progression, consistent with accumulating evidence that anomalies of mitosis are a significant cause of microcephaly. Remarkably, analyses of mitotic cells in static images and time-lapse imaging of Pals1-deficient progenitors revealed the emergence of internalized cells with nuclei inside of mitotic cells. This unusual cellular behavior mimics entosis, which is cell cannibalism utilized by tumor cells to engulf live neighboring cells for pro- or anti-tumorigenic purposes. It is unknown whether this extraordinary cellular event can be pathogenic in other diseases such as microcephaly. Therefore, the mouse model with an entosis-like process in its cortical progenitors will provide important new insights into the pathogenic mechanisms of microcephaly. Our preliminary study demonstrated that cell-in-cell (CIC) structures represent a dynamic and mobile cellular entity that is highly associated with lengthened mitosis and abnormalities in cytokinesis. As in tumor cells, ROCK inhibition completely abrogates CIC structures and restores the normal length of mitosis. Furthermore, we detected a striking increase of the P53 target, P21, in the Pals1 mutants and found that genetic elimination of P53 produces a remarkable rescue of cortical size along with substantial reductions of CIC structures and cell death. These observations lead us to hypothesize that Pals1 loss induces CIC pathology responsible for mitotic defects that compromise genomic content and cortical cell viability through the abnormal activation of Rho-ROCK and p53. To test this, we will determine the biogenesis, maintenance, and elimination of CIC structures and how they impact mitosis and subsequent genomic integrity and fate of cortical cells (Aim1). Next, we will study how Pals1 deletion/reduction causes entosis through abnormal Rho-ROCK activation (Aim2). Finally, we will delineate the effect of P53 activation on CIC formation and Rho-ROCK regulation in Pals1 mutant progenitors (Aim3). The current study provides an important molecular and cellular clue as to how Pals1 mutation causes such a dramatic cortical phenotype as the complete absence of the cerebral cortex and hippocampus. Furthermore, for the first time, our study will establish that an entosis like process can occur in cortical progenitors, providing a novel pathogenic mechanism by which entotic cell cannibalism produces microcephaly.

NIH Research Projects · FY 2025 · 2023-04

Project Summary Women involved with the carceral system are more at risk of HIV infection, substance use disorders, and overdose than their community-based counterparts. Pre-exposure prophylaxis (PrEP) has emerged as a powerful HIV-prevention tool and in combination with Medication for Opioid Use Disorder (MOUD) could significantly impact HIV and overdose risk. Patient navigation holds strong potential to address multifactorial and complex barriers to PrEP and substance use treatment linkage and uptake for women involved in the carceral system. In tandem with patient navigation, eHealth has the potential to improve healthcare engagement for this group of women. As a multidisciplinary team with expertise in HIV-prevention implementation science research, technology enhanced, gender-responsive, and behavioral health focused (substance use, mental health and HIV prevention) interventions, and intervention development for individuals involved in the carceral system, we propose to examine the potential impact of using a technology-based adjunct to an existing navigation program to improve linkage to PrEP and MOUD for women leaving carceral settings in Philadelphia (PA). Intervention development and study design will be guided by our team’s pilot and on-going research, and the EPIS Implementation Framework. Study aims are to: 1) Assess women involved in the carceral systems’ perceptions of PrEP and MOUD and potential barriers to initiation and adherence; 2) Develop the PA-Links web-based app and assess usability and acceptability; and 3) Pilot the intervention to assess for promise of efficacy in engaging women involved in the carceral systems in PrEP and MOUD by integrating PA-Links into existing patient navigation. A survey of women involved in the carceral system and perceptual mapping analyses will be used in Aim 1 to guide proto-type app development and user testing in Aim 2. In Aim 3, we will examine the primary outcome of PrEP and MOUD service linkage and secondary outcomes such as PrEP and MOUD prescription/initiation, PrEP and MOUD adherence/persistence, perceptions and knowledge, and acceptability of the intervention. A Community Advisory Board will inform all aspects of the proposed study. This pilot study will significantly contribute to the field of HIV and substance abuse treatment by examining the potential impact of using technology based, tailored, gender-responsive interventions in adjunct with existing navigation in an urban, community-based health center. Study results will provide crucial information for a subsequent R01 proposal to rigorously evaluate an urgently needed HIV prevention intervention developed specifically to meet the unique cultural and social needs of high-risk women.

NIH Research Projects · FY 2026 · 2023-04

(PLEASE KEEP IN WORD, DO NOT PDF) This project responds directly to PAR-21-038; the PI changes direction from valvular heart disease to study acute kidney injury (AKI). Indeed, the PI is a new faculty member in the Division of Nephrology, Department of Medicine. The PI will apply her previous expertise in platelet structure to examine a new therapeutic approach to deliver mitochondria to injured kidneys and therefore stop the progression of AKI to end-stage renal disease. The PI is working closely with Dr. Daehn, an expert in kidney disease and mitochondrial function. In addition, she will be collaborating with Dr. Brestoff, an expert in mitochondria transplantation, to validate the proposed experimental approach. AKI is a critical health condition characterized by a sudden decline in kidney function. It occurs in approximately 20%-30% of hospitalized patients. In the US, AKI is leading to high morbidity and mortality. Although AKI encompasses various etiologies, tubular injury is an early and decisive step in AKI. During hypoxia, proximal tubular epithelial cells (PTECs) undergo oxidant stress, mitochondrial damage, protein synthesis inhibition, and growth arrest. Non-treated AKI can progress to chronic kidney disease (CKD) and end-stage renal disease. Renal replacement therapy is necessary for patients with a survival of only 10%. Currently, there are no pharmacological or preventive strategies available to reverse or reduce the occurrence of severe AKI or to stop its progression to chronic kidney disease and end-stage kidney, emphasizing the need for new therapeutic strategies in this area. The central hypothesis in this application will test an innovative approach to treating kidneys using mitochondria transplants to prevent AKI. This hypothesis will be tested in one Specific Aim; To examine a new delivery approach of competent mitochondria to isolated proximal tubular epithelial cells using naked mitochondria or encapsulated mitochondria.

NIH Research Projects · FY 2026 · 2023-03

Project Summary In aging neurons, the accumulation of key misfolded proteins into aggregates is a hallmark of many neurodegenerative diseases. For example, pathological forms of TDP43 become mislocalized to the cytoplasm and accumulate in aggregates in frontotemporal dementia (FTD) and other Alzheimer Disease-related Dementias (ADRD). Highly intricate networks of enzymes called molecular chaperones combat these processes. In ADRD, it is unknown how the chaperone networks fail against pathological forms of ADRD proteins such as TDP43, FUS, and TAU. A major challenge to studying chaperone networks is the combinatorial complexity. The canonical Hsp70 network consists of 54 Hsp40, 12 Hsp70, and 16 Hsp110 gene variants, creating a landscape of 12,155 possible protein expression combinations. Unique combinations of the Hsp40-Hsp70-Hsp110 proteins are hypothesized to confer specificity for different misfolded proteins in the complex human proteome. This hypothesis is widely accepted but it has never been directly tested due to technical limitations. This NIH K99/R00 proposal outlines a plan to directly test this hypothesis by building the first exhaustive map of a chaperone network against the ADRD-associated proteins TDP43, FUS, and TAU. To achieve this goal, aim 1 will leverage a new genetic technique developed by Dr. Edward Barbieri to express and study all 12,155 possible combinations of the human Hsp40-Hsp70-Hsp110 network in yeast models of ADRD. The chaperones identified as having activity against TDP43 in yeast will be further studied in aim 2 using human cells and in vitro assays. With human cell models, Dr. Barbieri will study the effect of the TDP43-active chaperones on cytoplasmic TDP43 aggregation and assess if the chaperones restore native TDP43 function in mRNA splicing in both HEK-293T cells and neurons. Using in vitro biochemistry, he will measure chaperone activity for prevention and reversal of TDP43 aggregation. During the R00 phase in aim 3 Dr. Barbieri will apply the chaperone network screen to study the TAU aggregation and he will expand the chaperone networks studied in yeast by including Hsp40 pairs and small HSPs. Lastly, Dr. Barbieri will combine the skills he learns during the K99 phase to develop a screen for combinatorially overexpressing all 194 human chaperones directly in human cell models of ADRD to study proteostasis networks in the native context. Together, the experiments outlined in this proposal will identify key chaperones as therapeutic targets for ADRD. Dr. Barbieri will perform the K99 phase mentored in the Shorter lab at the University of Pennsylvania, a world class biochemistry lab with expertise in the study of chaperones and ADRD. This is an ideal training setting for Dr. Barbieri to acquire new skills in biochemistry. Furthermore, Dr. Barbieri assembled an advisory committee to provide expertise in ADRD and formal training in the human cell assays. The new skills will complement his current expertise in molecular genetics, and his outlined training plan will provide the necessary preparation for Dr. Barbieri to progress to his goal of an independent position.

NIH Research Projects · FY 2026 · 2023-03

High levels of food motivation among young children are heritable, track over time, and associated with elevated risks of unhealthy eating and obesity. Despite significant growth of family-based obesity prevention efforts, the evidence base is remarkably scant on parenting highly food motivated children to prevent obesity and poor dietary outcomes. The need to move away from a “one-size-fits-all” approach to food parenting is particularly important to ensure that the evidence base and the recommendations for obesity prevention efforts that stem from the research generalize to all U.S. families. Our goal is to generate robust basic science evidence on parenting highly food motivated children to prevent excessive dietary intakes and body mass index (BMI) gains during the preschool years. Using a prospective cohort design, we propose to follow 205 caregiver/child dyads at two points over 18 mo as children transition from preschool to elementary school, when significant numbers of children begin to experience problems of poor diet quality and obesity. We will recruit children with varying food motivation to understand whether highly food motivated children have different needs than other children. We propose a multi-method approach using state-of-the-art measures, including ecological momentary assessment, to comprehensively investigate the amount, types, and consistency of food parenting practices (i.e., specific, goal-oriented behaviors) needed to prevent food motivated behaviors, excessive dietary intake, and BMI gains in children. Specifically, we propose to comprehensively evaluate the role of structure (i.e., theoretically supportive) and its differentiation from more coercive types of food parenting control. Aim 1 will characterize multi-dimensional eating behavior profiles and obtain evidence of validity via associations with observed food motivated behaviors, excessive dietary intakes, and BMI gains over 18 mo. Aim 2 will evaluate the amount, type(s), and consistency of food parenting structure needed to prevent food motivated behaviors, excessive dietary intakes, and BMI gains among highly food motivated vs. other children over 18 mo. Aim 3 will evaluate whether structured food parenting practices are more beneficial in the context of global approaches to feeding or feeding styles that involve high levels of control (i.e., authoritarian) vs. more balanced styles (i.e., authoritative). Findings will 1) generate a basic science evidence base on food parenting approaches for highly food motivated children that currently does not exist, 2) provide greater specificity for family-based prevention efforts, and 3) increase the generalizability of research on food parenting in obesity prevention.

NIH Research Projects · FY 2025 · 2023-02

PROJECT SUMMARY The overall goal of this K01 award is to provide Gabriella McLoughlin, PhD, MS, with the training and mentorship to establish an independent program of research focused on improving implementation of Policy, Systems, and Environmental (PSE) interventions for childhood obesity and cardiovascular disease prevention. Childhood obesity disparities have been exacerbated in recent years, such that children from racial/ethnic minority and low-income backgrounds are most at risk. Multiple factors, such as increased food insecurity resulting in a low-quality diet, contribute to this increased threat. Childhood obesity tracks into adulthood, increasing the risk of cardiovascular disease, and warranting sustainable interventions for youth to address social determinants of health. Since children and adolescents consume over half of their energy intake at school, providing free nutrient-rich breakfast and lunch to low-income youth through Universal School Meals (USM) is a key PSE approach to addressing obesity disparities. Adoption of USM is linked with reductions in obesity risk for low-income students, improved diet quality, reduced food insecurity, and enhanced academic achievement outcomes. Unfortunately, schools adopting USM report consistent challenges to its implementation such as low student participation and financial difficulty in adhering to Federal nutrition requirements. Programs and policies designed to mitigate health disparities for obesity cannot make the most impact if they are not reaching their target population. Because the federal reimbursement rate for schools is tied to the number of students receiving school meals, maintaining participation is critical to making USM financially feasible. The purpose of this K01 award is to conduct an implementation mapping procedure with the School District of Philadelphia (SDP), resulting in the development and testing of an equity-focused implementation strategy for USM. This will be achieved through three aims: 1) Investigate determinants of USM implementation through a health equity lens; 2) Develop and tailor a USM implementation strategy through a community-engaged approach; and 3) Evaluate the implementation and student health outcomes of the implementation strategy. To accomplish these aims and prepare for a larger implementation effectiveness trial, the applicant will receive rigorous formal training in 1) assessing implementation determinants grounded in health equity 2) community-engaged implementation mapping, 3) randomized controlled trials, and 4) cost analysis of implementation under the mentorship of Drs. Jennifer Fisher, Omar Martinez, Resa Jones, Ross Brownson, Alex Dopp, and Shiriki Kumanyika. Temple University is the ideal setting for this work given a longstanding relationship with the local community, facilitating Dr. McLoughlin's collaboration with SDP and completion of the study aims through a community-engaged approach. This K01 award holds significant potential for public health impact and will enable Dr. McLoughlin to establish a rigorous program of research to improve PSE implementation and reduce obesity and cardiovascular disease risk in vulnerable populations.

NIH Research Projects · FY 2026 · 2023-02

When entering college, many students plan to major in STEM fields with the goal of attending medical school. However, many of these students realize that while they are passionate about science, clinical careers are not appealing. Undergraduate students may not be aware of the many research careers available to them. One way to provide exposure to these careers is via undergraduate research experience, which improves retention of students in STEM disciplines and the pursuit of advanced degrees. Given the benefits of undergraduate research experience, Temple University has programs that encourage undergraduate participation in neuroscience and psychology research, such as the MiNDS program and Independent Study credit options. However, these programs either require or prioritize students who can give a 2-year commitment. Thus, transfer students, students who are late to find their passion for research, and students who do not know about or feel comfortable asking about research opportunities until later in training, often miss out on the participating in undergraduate research. Additionally, Independent Study, which provides most research opportunities to undergraduates, is compensated via course credit, which can be an impediment for students who need to earn money to support their education (a large portion of Temple undergraduates). We started the Building Research Independence by Developing Goals and Hands-on Experiences (BRIDGE) program in 2021 to address these barriers. BRIDGE targets neuroscience and psychology majors who have not had the opportunity for research experience prior to their last (typically senior) year of college. BRIDGE provides scholars with paid summer research experience, individualized mentoring, and networking and professional development opportunities. This R25 grant would increase the support for the existing BRIDGE program, allowing students to work full time over the summer and providing travel funds, options that are not currently available. This funding would also support a near-peer mentoring program where doctoral students would mentor BRIDGE scholars. Near-peer mentoring positively affects both mentees and mentors by building community. Overall, the BRIDGE program will demystify the research process and provide more students with tools to pursue biomedical careers.

NIH Research Projects · FY 2026 · 2023-02

Project Abstract/Summary As a health services researcher and Assistant Professor at Temple University, my long-term career goal is to become a leader in preference-based, patient-centered outcomes research to inform decision making for patients with end-stage lung disease, specifically about transplantation decisions. Clinical trials have shown that decision aids improve the quality and efficiency of decision making, improve comprehension and decrease decisional conflict. Despite these benefits, few decision aids have addressed the complexity of lung transplant. A critical barrier to the development of decision aids is the inability to inform and communicate to patients and caregivers the impact of transplant on quality of life (QoL). Thus, to address these knowledge gaps and pursue my long-term career goals, I will investigate patient priorities for QoL outcomes in the context of lung transplant decision making to inform the development of a decision aid. Three Specific Aims will accomplish this objective. AIM 1: Describe priorities for QoL outcomes and their impact on decision making among patients (n=30-40) and caregivers (n=30-40) at each phase of lung transplant, and clinicians (n=10-15). AIM 2: Measure QoL, priorities for outcomes, and willingness to trade between quantity and quality of life among a prospective cohort of patients (n=200) and caregivers (n=200) considering lung transplant using health-related quality of life measures, best- worst scaling tasks, and treatment tradeoff exercises. AIM 3: Develop and assess the feasibility and acceptability of a decision aid among patient and caregiver dyads (n=30) considering lung transplant. To accomplish these aims, I propose a mixed method approach. Qualitative methods include semi-structured interviews conducted in cross-sectional samples across transplant phases (Aim 1). Quantitative surveys and the feasibility/acceptability study will be collected among patients with end-stage lung disease just starting to consider transplant, as well as their caregivers (Aims 2 and 3). A prospective cohort will be followed over time to identify differences across phases of decision making (Aim 2). Although I have had a productive start to my career, I require further training to execute these Aims, accomplish these goals and achieve independence as an investigator. Specifically, I have identified four short-term career objectives. I will: 1) lead a community-engaged research effort inclusive of patients, caregivers and clinicians; 2) design and implement qualitative and mixed methods approaches to collect and analyze patient experience data; 3) obtain formal training in theory of decision making, interpersonal health communication and risk communication; and 4) acquire domain expertise to apply these skills in the context of lung transplant. I have designed a career development plan to achieve these objectives that includes mentorship from a highly accomplished group of researchers who are dedicated to my career development and experts in decision making theory, decision aid design, health and risk communication, qualitative and mixed methods, and lung transplant. These mentors have long histories of extramural funding and mentoring. In addition to mentored instruction, I will complete didactic coursework and intensive professional trainings.

NIH Research Projects · FY 2026 · 2023-01

The control of platelet function and coagulation is a fine balance between activation and inhibitory mechanisms. Platelets become rapidly activated by multiple receptors agonists and have a central role in thrombosis in acute coronary syndromes, and other disease states. Similarly, naturally occurring anticoagulants are critical in preventing fibrin generation and thrombosis. Fivemembers of theprotein disulfide isomerase (PDI) family of enzymes, PDI, ERp57, ERp5, ERp72 and ERp46 potentiate activation of IIb3 and thrombosis.We discovered atransmembrane member of the PDI family found to inhibit activation of IIb3 and member of the PDI family in platelets, TMX1, which thrombosis. TMX1 is the first acts by a novel mechanism of oxidizing thiols to disulfide bonds and is the last checkpoint inhibitor of the platelet activation pathways that lead to conformational changes in IIb3 and fibrinogen binding. The prothrombotic PDIs are secreted from platelets and endothelial cells and support fibrin generation at the site of vascular injury. We found that TMX1 is expressed on platelets and endothelial cells but, negatively procoagulant in contras to the other PDIs, TMX1 regulates fibrin generation. One mechanism by which TMX1 inhibits coagulation is by limiting the effect of endothelial cells and platelets. We propose to study t vascular TMX1 as a dual negative regulator of platelets and coagulation by addressing the following Specific Aims. We will characterize 1. the role of TMX1 in thrombus formation; 2. the mechanism of inhibition of IIb3 activation by TMX1; 3. the effect of TMX1 on the other platelet PDIs, and on other platelet surface substrates. A principal technique used will be the laser-induced injury model of thrombosis. We will study the mechanism by which TMX1 negatively regulates coagulation. To determine the underlying mechanisms by which TMX1 inhibits platelet function we will integrate a platelet knockout mouse model with mass spectrometry-based identification of functional cysteines. This proposal will determine the mechanisms by which TMX1 works, and how TMX1 counterbalances the PDI enzymes that support activation of IIb3. Characterization of the negative regulatory role of TMX1 will provide novel insight into how the network of PDI enzymes regulate thrombosis. Studies on how TMX1 maintains the balance between thrombosis and hemostasis will elucidate optimal ways to promote hemostasis and inhibit thrombosis and provide a basis for studying TMX1 in disease states.

NIH Research Projects · FY 2026 · 2023-01

SUMMARY Myocardial infarction (MI) is a major cause of morbidity and mortality worldwide. Therefore, seeking new therapeutic approaches is still the priority in the field. Recent studies have shown that stimulation of lymphatic growth (lymphangiogenesis) after ischemic heart disease improves cardiac function and attenuates adverse cardiac remodeling, and cardiac lymphangiogenesis likely improves heart repair after injury by improving the clearance of inflammatory response. Furthermore, my published work has unraveled a novel additional mechanism that an “active”, pro-survival paracrine signaling (lymphoangiocrine, Reelin) from lymphatic endothelial cells promotes cardiomyocyte (CM) survival after injury, a result challenging the traditional view that lymphatics function as a “passive” route for fluid transport. However, the detailed mechanism/s by which increased lymphangiogenesis improves heart repair is not yet fully understood. Our exciting preliminary data suggest that lymphangiogenesis improves cardiac repair requires both, cardioprotective lymphoangiocrine signals and efficient lymphatic immune clearance function. This proposal is set out to use a various valuable animal models to determine these functional roles of cardiac lymphangiogenesis during cardiac repair. In the Aim 1 of the proposal, we will determine the molecular pathways and targets how cardiac lymphangiogenesis improve CM survival after MI. In the Aim 2 of this proposal, we aim to use a variety of lymphatic loss- and gain- of function mouse strains to characterize in details the lymphatic functions during cardiac repair, and determine the therapeutic potential of these optimal effects in treatment of ischemic disease. These fundamental findings will impact diverse fields including lymphatic biology and cardiovascular research, with an ultimate goal to identify novel targets for the treatment of cardiovascular diseases.

NIH Research Projects · FY 2026 · 2023-01

Title: LysoPI/GPR55 pathway promotes endothelial activation, vascular inflammation and atherosclerosis Atherosclerosis and its complications are the leading cause of morbidity and mortality in the US. Novel antiinflammatory therapies are urgently needed to inhibit atherosclerosis. The goal of this project is to determine the roles and mechanisms underlying lysophosphatidylinositol (lysoPI) promoted endothelial cell (EC) activation and atherosclerosis. We published numerous papers on EC activation, atherosclerosis and lysophospholipids (lysolipids). Our strong preliminary data and publications show that: 1) we reported a new concept that most lysolipids are new conditional classical damage- associated molecular pattern (DAMPs) that do not bind to classical DAMP receptors but bind to their own receptors to stimulate inflammation; 2) our metabolomics data showed that lysoPI is significantly increased in the aortas and plasma of ApoE-/- mice fed with high fat diet (HF) for three weeks; 3) lysoPI specifically bind to its receptor, G protein coupled receptor 55 (GPR55), and activates primary human aortic endothelial cells (HAECs) by upregulating EC adhesion molecule ICAM-1; 4) Mechanistically, lysoPI induces generation of mitochondrial reactive oxygen species (mtROS) in HAECs and induces proinflammatory proteinarginine methyltransferase 1(PRMT1) activity; 5) our RNA-Seq data showed that significantly different from that induced by lysoPI's relative lysoPC, lysoPI induces sustained EC activation by upregulating DAMP receptors inflammasomes/caspase-1 and proinflammatory secretomes; 6) we established ApoE-/-/GPR55-/- DKO mice; and DKO mice have significantly decreased atherosclerosis in comparison to that in ApoE KO mice fed with HF for 12 weeks; and finally, 7) we generated EC-specific GPR55 KO mice and PRMT1 KO mice. Based on our strong preliminary data and publications, the central hypothesis to be tested is that early hyperlipidemia-induced lysoPI stimulates and sustains aortic EC activation via a GPR55-PRMT1 pathway, proinflammatory monocyte (MC) recruitment, thereby contributing to atherosclerosis. We will test this hypothesis using three aims. Aim 1 will determine expression and function of lysoPI/GPR55-PRMT1 pathway in HAECs activated by hyperlipidemic stimuli and in aortas of ApoE-/- mice (relevant studies). Aim 2 will examine the mechanisms by which lysoPI/GPR55 induces sustained aortic EC activation via inducing PRMT1 upregulation-mtROS generation, which further prolongs EC activation (mechanistic studies). Aim 3 will determine whether inhibition of lysoPI/GPR55 and PRMT1 in EC (EC-specific KO) and other vascular cells (global KO) would decrease atherogenesis in ApoE-/- mice (therapeutic studies).

NIH Research Projects · FY 2026 · 2022-12

Down syndrome (DS) is a congenital condition resulting from partial or complete triplication of human chromosome 21. Virtually all the subjects with DS develop widespread neuropathology including amyloid- neuritic plaques, synaptic dysfunction and neurodegeneration, reminiscent of Alzheimer's disease. While the extra copy of amyloid- precursor protein (APP) on chromosome 21 is thought to play a major role in the development of this type of pathologic phenotype in DS, the underlying mechanisms responsible for these changes and their contribution to neurodegeneration are still elusive. Compelling evidence supports the hypothesis that physiological cellular proteostasis is of critical importance for neuronal health and that the mammalian target of rapamycin (mTOR) is a master regulator of this vital cellular function. However, while we know that the system is altered in DS we do not know whether it plays a functional role in the pathogenesis of DS and the onset of neuropathology. In our preliminary data we found that compared with healthy matched controls mTOR is hyperactive in selected brain regions of DS patients. Importantly, we observed that in the same subjects the dysfunction directly correlates with the pathology. Moreover, we show that mTOR is altered in the brain of a well- established mouse model of DS, the Ts65Dn mice, at an early stage of the phenotype, and associates with biochemical evidence of cell loss, suggesting a link between mTOR, neuropathology and neurodegeneration in DS subjects. Taking into consideration the scientific rigor of the previously published literature together with our recent findings we now propose a novel working hypothesis: alteration of mTOR signaling pathway is responsible for the onset of the neuropathologic DS phenotype and represents a novel and viable therapeutic target against it in DS subjects. In this proposal, we will assess the temporal relationship between dysregulation of mTOR in the brain of DS patients and the development of the neuropathology. We will then focus on investigating early events responsible for this dysregulation in the same DS subjects. Next, to prove its direct role in the pathogenesis of the syndrome, we will study the effect that modulation of mTOR activity and expression levels has on behavior impairments and neuropathology using in vivo models of DS. The results of our proposed studies will elucidate early changes and the functional consequences of altered proteostasis secondary to dysregulated mTOR in the development of the neuropathologic phenotype in individuals with DS. Importantly, our findings have the potential to identify new therapeutic opportunities for delaying its onset and /or halting its progression.

NIH Research Projects · FY 2025 · 2022-09

Project Summary For the past decade, the use of opioids has risen dramatically in the United States and the disproportional increase in opioid dependence and overdose death has led to the current opioid crisis. Although different measures have been taken to reduce opioid overutilization for pain management, opioid use in clinics continues leading to dependence and overdose. In there is a compelling need for non-opioid use of pharmacological addition, for the significant number of people with opioid use disorder, pharmacological therapies to complement current treatments for opioid disorder. A major challenge is to develop new treatment strategies that can attenuate the rewarding aspects opioids while preserving their powerful analgesic properties. endocannabinoid (eCB) system serves as a potential target for the development of new treatments as a complement to opioid based treatments. Several lines of evidence suggest The functional interaction between the opioid and the eCB system at the level of neurochemical, neuroanatomical and molecular pathways. Our preliminary results find that indirectly enhancing levels of the endocannabinoid 2- AG levels through pharmacological inhibition of its catabolic enzyme, monoacylglycerol lipase (MAGL), attenuates the rewarding effects of morphine, while maintaining its analgesic effects. In this proposal we will dissect at a circuit, synaptic and molecular level how elevated 2-AG attenuates opioid reward. Recent studies have underscored the role of local GABAergic neuronal inputs from the rostromedial tegmental nucleus (RMTg) in regulating the ventral tegmental area (VTA), a key dopaminergic brain region involved in opioid reward. Opioids are thought to act by disinhibiting RMTg inhibition onto VTA dopamine neurons by activating presynaptic mu opioid receptors (MOR), subsequently increasing dopamine cell firing and nucleus accumbens (NAc) activity that drives reward. However, little is known about how cannabinoid receptors (CB1R) and MORs signal and crosstalk at these key synapses. Aim 1 will examine 2-AG mechanisms in the VTA on opioid reward behavior and its effect on NAc dynamics. Aim 2 will examine the role of CB1R and MOR in the RMTg→VTA projection on opioid reward behavior and NAc dynamics. Aim 3 will examine synaptic and molecular mechanisms of CB1R and MOR crosstalk to determine how enhancing 2-AG levels leads to blunted opioid reward.

NIH Research Projects · FY 2026 · 2022-09

Asian Americans (AAs) represent approximately 7% of the U.S. population but account for over half of all Hepatitis B virus (HBV) infections nationally. Among the estimated 2.4 million Americans living with HBV, 58% are AAs, who exhibit the highest HBV prevalence across U.S. population groups. Despite recommendations from the CDC and U.S. Preventive Services Task Force to screen individuals at elevated risk, an estimated 68–75% of AAs remain unscreened and undiagnosed. Underdiagnosis has been linked to limited engagement with healthcare services, unfamiliarity with the healthcare system, and difficulties navigating available health information. Most existing studies have not fully examined how multiple layers of influence affect HBV screening and liver disease treatment. Preliminary findings among Chinese, Korean, and Vietnamese populations suggest that screening and care gaps stem from a range of individual and clinical factors. These factors contribute to delays in diagnosis, reduced services to treatment, and worsening liver disease. This study aims to identify and analyze individual and clinical factors that influence HBV-related liver disease management and outcomes. Using an adapted Socio-Ecological Model, the project will build upon a 20-year Regional Cancer Network and established collaborations with healthcare providers and local organizations in the Philadelphia, New Jersey and New York City metropolitan areas. Employing a mixed-methods approach, the research team will: Aim1. Analyze the longitudinal relationship between individual-level factors and HBV screening and liver disease management among 2,000 Asian American participants; Aim2. Evaluate the influence of organizational-level factors within clinical settings on screening rates and care continuity; and Aim3. Examine how patient-level behaviors and clinical care practices influence HBV screening and treatment outcomes. An integrative analysis will determine how factors at individual and clinical levels jointly impact engagement with HBV care. This will be the first longitudinal study to examine HBV-related liver disease management in high-risk Asian American subgroups, including those with early-stage liver cancer. The study findings are expected to support the development of targeted strategies to improve the quality of HBV and liver disease care for all populations at risk, contributing to national efforts to reduce the burden of chronic liver diseases.

NIH Research Projects · FY 2024 · 2022-09

Chronic obstructive pulmonary disease (COPD) is a major cause of mortality and morbidity in the United States and growing cause of chronic disease globally. At present the treatment options for COPD are limited and new therapies are needed to treat/prevent progression of COPD. Environmental factors and host response leading to imbalance in the ratio of oxidants to antioxidants, and proteases to antiproteases leading to chronic inflammation and tissue destruction is thought to play crucial role in development and progression of COPD. Quercetin is a plant flavonoid and has potent antioxidant and anti-inflammatory properties. Quercetin supplementation decreased markers of oxidative stress and inflammation in the plasma of patients with another chronic lung disease, pulmonary sarcoidosis. Quercetin also decreased both oxidative stress and lung inflammation in a preclinical model of chronic obstructive pulmonary disease (COPD). However the impact of quercetin in altering biological signatures in COPD subjects is yet to be determined. The proposed study is conducted in two phases, R61/R33 to define and confirm the biological effects of quercetin in COPD, bioavailability, safety, identification of the appropriate biological signatures which reflect clinical outcomes in COPD. This study will also define the optimal dose of quercetin required for altering biological signatures favorably. This information is necessary to conduct the future clinical trials with quercetin in COPD patients. In R61 Phase, there will be 15 study participants with moderate COPD with FEV1 ranging between 45 and 70 % of predicted and they will be randomized to receive either placebo or quercetin 2000 mg/day for 6 months. In R33 phase there will be 35 COPD subjects, who will be randomized to receive placebo, or one of the 3 quercetin doses, 500, 1000 or 2000 mg/day. Outcome of R61 phase will determine the feasibility of R33 Phase. In both R61 and R33 phases, all subjects will be characterized using a validated symptom questionnaire, medication and smoking histories, analysis of co-morbidities, and physical examination. All subjects will be asked to go on a low quercetin diet 7 days prior to start of quercetin or placebo supplementation to until the end of trial. Blood and bronchoalveolar lavage (BAL) will be collected after 7 days washout period (run-in) and after study drug treatment. Blood will also be collected at 3 months of study drug treatment. Plasma and BAL quercetin levels, markers of inflammation and oxidative stress in blood and BAL will be measured. Pulmonary function, and blood profiles (CBC and CMP) will be performed at run-in and at 3 and 6 months of study drug treatment to assess the safety. Results from these studies will provide information on biological endpoints, safety, bioavailability, and quercetin dosage required to carry out large clinical trials examining the efficacy of quercetin in COPD patients.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY/ABSTRACT While clinical use of therapeutic proteins has grown exponentially in recent decades, utility is often limited by unfavorable pharmacokinetics (PK), mediated by rapid elimination. One approach to overcome this limitation is half-life extension (HLE), achieved by attachment of biotherapeutics to polymers (PEG), plasma proteins (albumin, IgG), and blood cells. There is a paucity of data describing the impact of properties of cargo proteins and HLE-conferring ligands on PK of proteins tested using these strategies. One area where HLE has the potential to advance clinical therapy is acute, life-threatening, thrombosis (pulmonary embolism, ischemic stroke, etc.). The only pharmacologic treatment available for treatment of this condition is infusion of plasminogen activators, which have an unfavorable pharmacologic profile, mediated by a half-life of minutes, rapid inactivation in plasma, and severe adverse effects (e.g. hemorrhagic transformation). Pilot data demonstrates that conjugation of a derivative of urokinase selectively activated in thrombin-rich pathological thrombi (scuPA-T) to an albumin-binding nanobody (Nb) leads to PK that is identical to RBC-binding scuPA-T, an approach pioneered by Drs. Muzykantov and Cines, albeit with distribution in the plasma rather than the cellular fraction of blood. The central hypothesis of this proposal is that coupling of scuPA-T to albumin-binding ligands will provide prolonged circulation, mediated by the neonatal Fc receptor (FcRn), and selective thrombolysis of pathological thrombi, sparing hemostatic plugs from lysis. In the mentored stage, the impact of properties of the cargo drug on FcRn-enabled HLE will be identified, using FcRn knockout mice to directly elucidate the FcRn-mediated component of HLE conferred by albumin binding (Aim 1; K99). Based on pilot data, safety and efficacy of albumin-binding scuPA-T will be studied in a mouse model of pulmonary embolism. These results will be used to develop a predictive modeling platform that will be used to for further engineering and PK optimization of HLE-scuPA-T constructs (Aim 2; K99). With an eye on mechanistic and translational advancement of this strategy, the role of albumin-binding affinity on the PK of scuPA-T will be defined, using a newly developed library of nanobodies (Aim 3; R00). Additionally, thrombin-cleavable HLE ligands will be devised, permitting selective release of scuPA-T in thrombi, improving diffusion into clots and lysis (Aim 3; R00). Overarching themes of this proposal include identification of critical features of albumin-mediated HLE and mechanism-based modeling to guide optimization and reengineering of protein therapeutics. A mentoring team has been identified with expertise spanning the areas of research in this grant, namely, thrombosis and hemostasis, antibody engineering, and mathematical modeling of biological systems. Mentored research will be conducted at the University of Pennsylvania, which has a highly collegial and collaborative faculty and extensive resources available to conduct the proposed research. This proposal is geared towards gaining the expertise necessary to be successful as a tenured faculty member conducting high quality biomedical research.