Temple Univ Of The Commonwealth

NIH Research Projects · FY 2025 · 2025-07

1 This proposal aims at solving the unmet biomedical needs in the treatment of castrate-resistant 2 prostate cancer (CRPC) which essentially develops to be drug-resistant with ligand-independent 3 androgen receptor (AR) signaling. CRPC cells constantly upregulate c-Myc which was believed 4 to drive the CRPC malignancy and progression. The c-Myc mediated transcription in CRPC also 5 promoted the expression of AR splice variants (AR-V) that were truncated and lacked ligand 6 binding domains thereby endowing CRPC cells resistance to common AR antagonist such as 7 enzalutamide. Thus, c-Myc appears to be an important target for elucidating the AR signaling in 8 CRPC and for novel therapeutic development. Yet, there is a lack of intracellular tools or chemical 9 probes available to effectively target c-Myc, as it belongs to a typical basic helix-loop-helix (HLH) 10 transcription factor family which consist of flat, hydrophilic, and extended surface areas with 11 limited small molecule binding pockets and is considered “undruggable”. Although there have 12 already been c-Myc bound peptide fragments that can effectively inhibit c-Myc-Max dimerization 13 in vitro, most peptides discovered to date suffer from limited biostability and membrane 14 permeability, thereby largely limiting their applications towards in vitro only. We recently invented 15 fluorine-thiol displacement reaction (FTDR) and applied this to staple a diverse set of peptide 16 mimetics, which later displayed significantly enhanced cellular and nuclear uptake. Based on this 17 observation, we hypothesize that application of the FTDR stapling strategy to rationally designed 18 and c-Myc targeted peptide mimetics will result in a class of chemical probes with efficient cellular 19 and nuclear uptake and will efficaciously inhibit c-Myc mediated transcriptional activity in CRPC 20 cells and in vivo mouse xenografts. The first aim seeks to utilize FTDR to staple a H1 peptide 21 sequence designed to target the helix-loop-helix region of c-Myc and will thoroughly evaluate its 22 inhibition of c-Myc-Max heterodimerization and the suppression of c-Myc-mediated transcription 23 in CRPC cells, as well as the associated AR signaling particularly related to AR-FL and AR-V7. 24 The lead hit peptide mimetic that potently blocked the proliferation of CRPC cells, but not regular 25 prostate tissue cells will be eventually assessed in mouse CRPC xenograft models. The second 26 aim will exploit targeting at the C-tail leucine zipper (LZ) domain of c-Myc to directly block the 27 MYC-Max dimerization and will also explore the tandem stapling on the peptide mimetics for both 28 H1 and LZ domains. Completion of these research goals will result in a toolbox of c-Myc targeted 29 cellular probes, which will systematically dissect c-Myc mediated drug resistance and will facilitate 30 future therapeutic development for CRPC treatment or as adjuvants of AR antagonist.

NIH Research Projects · FY 2026 · 2025-07

Methamphetamine (METH) addiction lacks an approved medication, which is different from opioid, alcohol and nicotine addiction. The NIDA therapeutic pipeline for psychostimulant addiction is also limited compared to opioid use disorder. This indicates a need to identify new targets and strategies for METH use disorder (MUD). The neuroimmune system is one promising target. It sits at the crossroads of the immune and central nervous systems. Neuroimmune signaling in brain circuits related to addiction is dysregulated during METH exposure and abstinence, and this dysregulation facilitates METH dependence and relapse. Non-selective anti- inflammatory agents are effective in animal models of psychostimulant addiction but can disrupt normal immune function. We propose a more selective strategy, one which targets a specific element of the neuroimmune system. The element we selected is the chemokine system, namely a trio of chemokine receptors (CCR2, CCR5 and CXCR4) that are (1) expressed in brain circuits that underlie METH dependence and relapse; (2) dysregulated during METH exposure; (3) enhance multiple neurochemical correlates of METH addiction (e.g., dopamine and glutamate transmission); and (4) `druggable' receptors with FDA-approved antagonists and a mechanistically unique preclinical antagonist. The overall hypothesis to be tested is that chemokine receptor antagonists (CRAs) blocking three different chemokine receptors (CXCR4, CCR2, and CCR5) will reduce METH intake, reinforcing efficacy, and relapse in SA assays, counteract anxiety-like effects during METH withdrawal, and normalize dysregulation of chemokine-dopamine crosstalk after chronic METH intake. The CRAs proposed for characterization are not known to be abuse liable, mechanistically distinct, and positioned at different stages of the neuroimmune pipeline. Two of the CRAs (AMD3100, a CXCR4 antagonist and maraviroc, a CCR5 antagonist) are approved for other therapeutic indications and eligible for repurposing. R-103, a preclinical CRA, is mechanistically unique because it blocks multiple chemokine receptors (i.e., CCR2, CCR5, CXCR4), thus offering potential for enhanced efficacy. R-103 also has favorable safety and pharmacokinetic profiles, including rapid brain entry and persistent brain and plasma levels. Since the CRAs proposed for testing may have varying potencies, half-lives, and metabolic/PK parameters, we will also determine the causal role of the specific chemokine receptors in METH intake and relapse behaviors using shRNA knockdown of CCR2, CCR5 and CXCR4 in the mesolimbic pathway. In summary, our results will offer the first comprehensive information about the efficacy of CRAs and the influence of specific chemokine receptors on METH SA behaviors and neurochemical effects. By identifying the most effective CRA, and determining the chemokine receptor with the greatest influence, we expect to pave the way for developing effective chemokine-based medications for METH use disorder.

NIH Research Projects · FY 2026 · 2025-07

Project Summary/Abstract A post-genomic era is upon us, marked by the rapid availability of massive sequence data. Storing, retrieving, and analyzing such high volumes of raw sequence data, as well as alignment information, pose significant challenges to researchers in the field. Consequently, new methods are desirable to support efficient storage and retrieval of such data to meet the demand of the exponential growth in sequence data. Within this scope, we will develop a suite of novel computational methods for compressing sequence data. Our proposed research includes three specific aims to develop: (1) an error-bounded lossy compressor for quality scores in sequence files, (2) an adaptive compression strategy to achieve optimal performance, and (3) a high-throughput compression framework for efficient sequence compression. We will extensively evaluate the proposed approaches on real and synthetic datasets, and develop modularized software tools to facilitate the application of the proposed methods. With a well-defined research plan for developing innovative methodologies to compress sequence data, this project will establish a new paradigm of data compression to support efficient storage, retrieval, transfer, and analysis of the ever-growing DNA and RNA sequencing data.

NIH Research Projects · FY 2025 · 2025-06

Abstract The human neurotropic virus, JC virus (JCV), causes a devastating and fatal disease of the human brain known as progressive multifocal leukoencephalopathy (PML). JCV establishes an asymptomatic infection for more than 70% of the human population during childhood and resides primarily in the kidneys and B cells in a latent form. Every individual with immunocompromised conditions, including AIDS patients and those treated with immunosuppressive drugs, for example, Crohn's disease, multiple sclerosis (MS), psoriasis, and rheumatoid arthritis patients are at risk for reactivating JCV and developing PML. Reactivated virus carries more than 20% fatality rate, and even those who survive the disease are severely disabled due to brain damage caused by the viral infection and inflammation. Currently, there are no effective treatment options available for PML patients, nor are there animal models that manifest the infection and symptoms of PML. The only treatment option is restoring the immune system to clear the infection. Still, even this often leads to increased brain damage, resulting from a condition known as "immune reconstruction inflammation syndrome (IRIS)." Thus, to address the unmet medical need for the treatment of JCV infection and to improve the management of PML patients, there is an urgent need to understand the disease progression at the molecular level to develop effective treatment options. Such options will only be possible by understanding the molecular mechanisms governing the JCV biology, which is mediated by its critical regulatory proteins. JCV encodes a limited number of such proteins encoded by the alternative splicing of the viral early and late transcripts. We have recently discovered a novel regulatory protein of JCV, ORF4 protein, generated by the viral late coding region, and its expression was validated during the viral infection cycle by proteomics studies. ORF4 is a small protein (173-aa long) found to be the only JCV protein specifically targeting the promyelocytic leukemia nuclear bodies (PML-NBs) and induce their reorganization in the nucleus. PML-NBs are known to play critical roles in intrinsic and innate immune responses against various viral infections, suggesting that ORF4 protein, by altering their reorganization, inhibits the function of PML-NBs and thus significantly contributes to the progression of the JCV life cycle. Our novel discoveries further revealed that ORF4 protein differentially regulates the reorganization pattern of the four permanent members of the PML-NBs. Three members, including PML protein, Daxx, and ATRX, remain together in the same complex after reorganization, while the fourth member, Sp100, behaves differently. Collectively, all these novel findings provide us with a strong rationale to investigate further the functional consequences of this targeting in vitro and in vivo model systems and allow us to formulate our central hypothesis, which is that "JCV ORF4 protein inactivates the antiviral response function of the PML-NBs by altering their distribution pattern and thus significantly contributes to the progression of the viral infection cycle". To further examine our hypothesis, we propose two Aims: (i) Investigate the molecular mechanisms by which the ORF4 protein induces reorganization patterns of PML-NBs, employing in vitro tissue culture model systems, and (ii) further evaluate the functional roles of ORF4 protein by creating an innovative mouse model system.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY/ABSTRACT: More than 80,000 opioid overdose deaths were reported in the US in 2022. Medication for opioid use disorder (MOUD) significantly reduces the risk of hospitalization and mortality from opioid overdose, but MOUD is severely underutilized. In addition, there are significant geographic, racial, and ethnic disparities in MOUD use. Medicaid is the largest payer of treatment for opioid use disorder (OUD), with the potential to reach a large share of the population in need of treatment. In 2015, state Medicaid programs began to adopt the newly created Section 1115 “Institutions for Mental Diseases” (IMD) waivers. These waivers allow states to use federal funds for substance use treatment in residential facilities. To date, 33 states and the District of Columbia have adopted an IMD waiver. However, it is unclear whether waivers have an impact on MOUD use and retention. State Medicaid programs have broad discretion on services covered for substance use disorders, which may lead to significant variation in treatment across jurisdictions. A careful examination of waiver configurations to compare them across jurisdictions is needed, and this effort will be among the first. This Mentored Research Career Development Award (K01) proposes a community-engaged, mixed methods research to (1) characterize IMD waivers characteristics and provide a detailed description of waiver features in a standardized way that allows for comparison across jurisdictions, (2) quantitatively measure the impact of IMD waivers on MOUD-based treatment and health outcomes (e.g., MOUD initiation, retention, and overdoses) using Medicaid claims, and (3) qualitatively explore waiver implementation in the first 10 jurisdictions that adopted a waiver using in-depth interviews with stakeholders engaged in state Medicaid agencies, provider networks, and advocacy groups. Mixed methods will be used to triangulate findings from the aims and provide a comprehensive picture of the policy. The community-engaged component of this study will include an advisory board to provide ongoing input, including interpretation of results to translate study findings into practical knowledge for policymakers. Findings from this research will characterize persisting and potentially new gaps in access to MOUD that, if addressed, can substantially improve public health and well-being. The training and mentoring plans will build silks and expertise in (1) legal epidemiology methods to code and analyze waiver policies (Aim 1), (2) advanced causal inference methods using real-world Medicaid claims data and content expertise on opioid use disorders treatment to measure the impact of waivers on outcomes at the population level (Aim 2), (3) community-engaged and mixed methods approaches to investigate the implementation of waivers at the local level and to triangulate data from quantitative and qualitative aims guided by the perspective of stakeholders with lived experience. This award will provide a platform to launch an independent research program that will fill a critical gap in knowledge about policies to scale up MOUD use.

NIH Research Projects · FY 2026 · 2025-04

The `tranq dope' threat involving combination of xylazine and fentanyl has been highly publicized, but emerging evidence indicates that methamphetamine (METH) is increasingly being adulterated with xylazine. A recent toxicology study reported that greater than one-third of all human METH samples were co-positive for xylazine. These emerging trends of METH and xylazine misuse necessitate pharmacological studies to determine if, and how, xylazine worsens METH effects. Evaluating effects of xylazine on METH-induced behavioral alterations in preclinical studies is a first step in better defining METH-xylazine interactions and guiding public health responses to xylazine-METH co- exposure. To our knowledge, there is not a single preclinical study that has investigated effects of xylazine on METH-induced effects. For amphetamine, there is only a 1994 rat study, which showed that convulsions elicited by amphetamine are reduced by a high dose of xylazine. In our studies, we observed xylazine-mediated regulation of METH locomotor and self-administration (SA) behaviors in adult rats. Our data demonstrate that xylazine (1) dose-dependently reduces locomotor activation evoked by acute METH and (2) increases METH infusions under fixed-ratio ((FR-1) SA conditions. These results provide a foundation to begin defining the unique behavioral and molecular consequences of dual METH and xylazine exposure. We will now expand upon these exciting findings to test the central hypothesis that xylazine (1) enhances acquisition of METH SA under FR-1 conditions, (2) enhances METH reinforcing efficacy, and the motivation to work for METH, under progressive-ratio (PR) conditions, (3) exacerbates anxiety- and depression-like effects that are associated with abstinence from chronic METH exposure, and (4) enhances METH seeking behaviors reinstated by different factors (METH prime, cue, or xylazine itself). We will also determine impacts of XYL-METH co-exposure on biomarkers of METH addiction in the nucleus accumbens transcriptome using unbiased RNA sequencing. We hypothesize that targets such as VMAT, TAAR1, DAT, D1, D2, and tyrosine hydroxylase, which are known to contribute to behavioral effects of METH, may be further exacerbated by METH-XYL co-exposure. To best match human preparations and human intake of XYL and METH, the two drugs will be mixed (in the syringe), and rats will be allowed to self-administer the drug-drug combination. Follow-up studies will probe receptor mechanisms underlying xylazine-METH interactions, including α2-adrenoceptor activation or kappa opioid receptor activation by xylazine. This comprehensive analysis of polydrug exposure of METH in combination with XYL will provide the first preclinical information about how XYL affects METH intake, reinforcing efficacy, seeking behaviors, and the nucleus accumbens transcriptome.

NIH Research Projects · FY 2026 · 2025-01

Project Summary/Abstract: Small cell lung cancer (SCLC) is a recalcitrant cancer that will kill more than 30,000 Americans this year. Annual incidence is nearly equal to mortality rates, and even patients with earlier stage disease infrequently live for two years beyond their diagnosis. There is currently a desperate need for novel therapies for SCLC. Any condition leading to elevated levels of DNA damage will result in replication stress which in turn is a source of genomic instability and increased vulnerability to specific agents including ATR inhibitors and DNA damaging agents. SCLC is characterized by high levels of replication stress (RS) although the exact mechanism causing the increased RS in SCLC is poorly understood. Interestingly, I have demonstrated that low LMNA (a nuclear structural protein) is highly correlated with increased RS, and that SCLC cells have very low levels of LMNA. I have demonstrated that low LMNA leads to increased chromatin accessibility and increased R-loops (RNA DNA hybrids) due to promoter proximal pausing of RNA Pol-II. R-loops are highly associated with RS, and replication fork collisions with R-loops can even cause DNA damage. The first aim of this grant is to fundamentally understand whether low LMNA causes increased RS in SCLC, and if so to determine if the mechanism R-loop dependent. Secondarily, during the R00 portion of this work, I plan to target R-loops and LMNA utilizing R-loops stabilizing agents and LMNA inhibitors to augment ATR/DNA damage based therapeutic regiments we have demonstrated have greater efficacy in SCLC patients with increased RS. SCLC cancers are characterized as having strong neuroendocrine (NE) differentiation, and a high prevalence (~80%) of RB (tumor suppressor which suppresses G1/S transition) mutation in patient tumors. I have demonstrated that SCLC cells compared to other cancer types are highly sensitive to loss of G1/S transition facilitating targets (G1ST) including CDK2, E2F, Cyclin E and SKP2. This is unsurprising as many of these targets have been found to be synthetic lethal with RB mutation. However, I have demonstrated that RB proficient SCLC cells (no RB1/2 mutation or copy number loss) are still more sensitive to loss of G1ST, and this increased dependency is strongly correlated with NE differentiation. My second aim is to determine whether the increased dependence of SCLC on these G1ST is caused by RB mutation, NE differentiation, or a combination of both factors, and to determine the mechanistic underpinning of this interaction. During the R00 portion of this work I will target G1ST to augment standard of care regimens in SCLC. This work will provide deliverables as follows: 1) determine the mechanistic underpinning of increased RS in SCLC 2) a novel therapeutic approach targeting RS, LMNA and R-loops in SCLC 3) determine mechanisms behind increased SCLC on G1ST 4) a novel therapeutic approach targeting G1ST in SCLC.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY/ABSTRACT Binge drinking (BD) is defined as a rapid consumption of alcohol that brings blood alcohol content to .08% or higher, typically described as ≥4 drinks for women and ≥5 drinks for men per occasion. BD is associated with many adverse consequences, including accidents, physical and sexual assault, and alcohol poisoning, as well as increased risk for developing an alcohol use disorder (AUD). BD is highly prevalent among American college students, with past-month estimates of 30% or greater. College – in particular the first six weeks – represents a critical time for the onset and development of harmful alcohol use. Drinking patterns established early in college influence alcohol use in the years following, underscoring the need for research into the early detection and prevention of BD in first-year college students. Certain health behaviors – namely sleep and physical activity – have been shown to bidirectionally influence BD, though the direction of the relationship is equivocal in college students. Furthermore, both acute and recurrent BD have been associated with psychophysiological changes, including diminished heart rate variability (HRV), leading some researchers to conceptualize HRV as a biomarker of AUD. However, existing research is either correlational or describes the impact of drinking on HRV, rather than examining HRV as a risk factor for BD, despite conceptual support for this direction. Moreover, measuring health behaviors and psychophysiology with traditional self-report measures or laboratory paradigms poses methodological challenges such as biases in participant recall, high participant burden, low compliance, and limited ecological validity. In contrast, smartwatches – which can passively sense many health variables with minimal user input – are being increasingly employed in psychological and medical research as a way of mitigating these concerns. The proposed study will utilize passive sensing via smartwatches in a sample of 100 incoming college freshmen (two cohorts of 50) without a history of BD or AUD prior to the start of their first semester. Participants will complete a baseline assessment of their substance use history followed by a six-week smartwatch monitoring period. Throughout the semester, participants will complete weekly self-report measures of BD accompanied by periodic timeline follow-back interviews and urine ethyl glucuronide screening for validation purposes. Follow-up assessments will be completed at the end of the first semester (4 months) and second semester (9 months). Specific variables of interest are: physical activity (steps and logged workouts); sleep (duration, chronotype, and social jetlag); and HRV. Data collected during the smartwatch monitoring period will be examined as prospective predictors of BD onset and frequency, with an exploratory aim focusing on AUD onset. Results from this study will help identify novel biopsychosocial risk factors for BD during a critical period in the establishment of drinking trajectories, with implications for prevention of alcohol-related harm. In collaboration with an expert mentorship team, the applicant has developed a training plan to gain the skills needed for successful implementation of this study in addition to professional development.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY Drug addiction represents an enormous healthcare burden. To better understand its biological underpinnings, investigations of the transcriptional response to drugs of abuse have demonstrated lasting changes in gene expression throughout the brain’s reward circuitry. Historically focused on neurons, emerging evidence increasingly indicates that astrocytes are also involved in disorders of the nervous system, including addiction. Indeed, candidate genes in astrocytes have been identified and, furthermore, manipulation of astrocyte function has been demonstrated to influence rodent behavioral responses to cocaine administration. However, the astrocyte-specific transcriptome and its regulation following exposure to drugs of abuse has not yet been investigated. In my career, I will build an independent research program that investigates the astrocyte transcriptome and its regulation in the context of addiction. This proposal will elucidate the underlying molecular mechanisms governing astrocytic CREB’s regulation of cocaine addiction-related behavior. Under the mentorship of Drs. Eric Nestler and Paul Kenny, I will rescue neuronal hypoactivity with DREADDs stimulation and record astrocyte calcium transients during cocaine SA. With additional mentorship from Dr. Li Shen, I will determine sex-specific astrocytic histone H3K27me3 gene targets and downstream molecular and cellular consequences to increase the rewarding properties of cocaine through bioinformatic comparison of CUT&RUN- and RNA-Sequencing. Viral-mediated manipulation to key H3K27me3 demethylase (Kdm6a/UTX) will uncover the contribution of astrocytic epigenetic regulation to cocaine drug-seeking. My independent laboratory will investigate the transcriptional and epigenetic regulation of astrocytes in the context of drug abuse and addiction. These experiments will prepare me to examine and manipulate cocaine-induced transcriptional and epigenetic modifications in astrocytes in my independent research laboratory. In sum, the research proposed in this Career Mentored Award will increase our understating of how regulation of the astrocyte transcriptome modulates behavioral responses to drugs of abuse. More broadly, the added training afforded by this award will prepare me to launch an independent research program that investigates the regulation of astrocyte gene expression, at both the transcriptional and epigenetic level, and distinguish its contributions to addiction phenotypes.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY - ABSTRACT Central to the development of HF are derangements in cardiac metabolism and a shift in fuel utilization. A recent clinical report of arterial/coronary sinus metabolomics identified glutamine as the most abundant metabolite released from the failing heart (i.e., production of glutamine rather than catabolism). Glutamine metabolism is directly linked to bioenergetics, mitochondrial function, redox state, nitrogen balance, and the biosynthesis of nucleic acids and proteins. Furthermore, we’ve recently shown that glutaminolysis is necessary for myofibroblast differentiation and persistence, that underlies cardiac fibrosis. Coupled together, these findings directed us to investigate if cardiomyocyte (CM) glutamine production plays a pivotal role in fibroblast activation and cardiac fibrosis. In preliminary data generated for the current proposal, we found an appreciable level of CM glutamine production during stress. Further, we discovered a significant increase in the expression of glutamine synthetase (GS; glutamate➞glutamine; GLUL gene) and numerous glutamine transporters in models of HF and in HF patients. These findings support the concept of enhanced CM glutamine production, and excretion in the stressed heart. To test the role of GS in cardiac pathophysiology, we generated a CM- specific, tamoxifen-inducible knockout model (GSfl/fl x αMHC-iCre; GS-cKO) and subjected mice to pressure- overload-induced HF (TAC). Loss of CM glutamine synthesis preserved LV structure and function and strikingly decreased fibrotic remodeling. In summation, our results suggest paracrine metabolite signaling from CMs may be a primary mechanism for fibroblast activation in the diseased heart. In addition to changes in fibroblast activation and ECM deposition, we also noted changes in CM metabolism, including increased mitochondrial respiratory capacity. In this project we will test the central hypothesis that targeting cardiomyocyte glutamine synthesis is a multipronged therapeutic strategy to increase bioenergetics, restore redox balance, and reduce fibroblast activation and fibrosis in HF. 1) We will examine if targeting CM glutamine synthesis reduces HF development, HFrEF and HFpEF models. 2) Determine if CM-derived glutamine is a signal/fuel for fibroblast activation, proliferation and myofibroblast differentiation/identity in the stressed heart. 3) Define the impact of targeting glutamine synthetase on cardiomyocyte metabolism, energetics, redox, and nitrogen balance. All models and methodologies for the proposal have been generated and validated. The proposed studies will allow for the first-time a causal examination of glutamine synthesis in cardiac physiology and disease.

NIH Research Projects · FY 2025 · 2024-12

Project Summary: The Society for Pelvic Research (SPR), founded in 2015, by and for career basic and translational scientists interested in normal function and benign disease states of the pelvic viscera and pelvic floor, is dedicated to cross-disciplinary basic and translational scientific interaction and trainee development. The society's primary mission is to promote the highest standards of basic and translational scientific research aimed at understanding the function and dysfunction of benign pelvic visceral and musculoskeletal systems through education, scientific discourse, interactions, and advocacy. This includes various conditions such as urinary and fecal incontinence, male and female sexual dysfunction, overactive and underactive bladder syndromes, colonic motility disorders, urologic chronic pelvic pain syndrome, and pelvic organ prolapse. These disorders are highly prevalent, particularly among our aging population, and exert significant negative impacts on the health, quality of life, and financial well-being of a substantial portion of the population. The SPR aspires to be the foremost professional organization for career basic and translational scientists and bioengineers interested in benign urogenital, distal gut, and pelvic floor research. It actively promotes multidisciplinary interaction, intellectual cross-fertilization, networking for collaboration, and career development through the regular dissemination of information via online resources, and, most importantly, through the annual SPR meetings. The objective of this R13 application is to request partial support for the 9th annual meeting of the SPR, scheduled to take place at Duke University in Durham, North Carolina, from December 5 to 7, 2024, with the Trainee Workshop Session occurring on December 5. An innovative immersive hybrid option will be provided. While there are other forums to present research findings, the SPR is the only society whose stated primary focus is to support the careers of basic and translational scientists that form the backbone for the discoveries that will eventually be implemented into clinical practice. At the 2024 meeting, each session, covering a diverse array of topics of interest to current and prospective future members of the SPR, will be unified by the overarching theme of aging (thus, our request for support from NIA), further emphasizing the distinctiveness of this conference. At the 2024 SPR Meeting, invited keynote speakers that are recognized leaders in our field will cover the influence of aging and age-associated conditions on the bladder and lower urinary tract function, in presentations entitled: “Age-relatedloss of bladdercontractility”, “Purine nucleoside phosphorylase as a target to treat age-associated lower urinary tract dysfunction”, “How aging affects the conversations between our gut and our brain”, and “Newtherapies for age-related erectile dysfunction ”. The SPR is committed to diversity and inclusion, as evidenced by more than 50% of positions on its Board of Directors (BOD) that are held by women. Our intention for this meeting, as with previous meetings, is to ensure the participation by underrepresented basic and translational scientists as invited speakers and attendees using targeted advertising, targeted invitations, and travel grants.

NIH Research Projects · FY 2026 · 2024-11

SUMMARY Most adults are latently infected with at least one of the two herpes simplex viruses HSV-1 or HSV-2. Upon reactivation these viruses can cause painful skin and mucosal lesions. Numerous patient populations with compromised immune systems or skin barrier function are at risk of frequent or severe disease and thus greatly diminished quality of life. Atopic dermatitis is the most common chronic inflammatory skin condition and is associated with diminish antiviral immunity in the skin through mechanisms that are poorly understood. Drug resistant HSV strains can develop especially in transplant patients. Thus, there continue to be a need for improved treatment strategies that encompass the host immune status. IL-1 and IL-36 are pleotropic cytokines that can promote innate and adaptive immune responses. We recently demonstrated that both IL-1 and IL-36 play important roles in limiting disease severity in a mouse model of reactivation-like disease caused by HSV-1. Pellino1 (Peli1) is a ubiquitin ligase involved in IL-1/IL-36 and TLR signaling. Studies of RNA viruses in vivo have generated diverging conclusions regarding beneficial and detrimental functions of Pellino1. Using our previously established infection model, we find that HSV-1, a DNA virus, causes greater mortality and larger lesions in Pellino1 knockout mice than wild type. In Peli1-/- mice, the virus disseminates rapidly through the skin epidermis and hair follicles; thus, Pellino1 clearly has one or more protective immune functions in keratinocytes and the epidermis. RNA-seq analyses revealed surprising mechanistic insight. Our newly developed hypotheses will be tested in keratinocyte specific conditional knockout mice using the established mouse infection model. Further validation of concepts will be obtained through use of human and mouse conventional and organoid-like cell cultures. We will define how the studied mechanisms may limit viral replication in keratinocytes by preventing, for example, cellular entry and exit of mature viral particles. Improved insight into these antiviral mechanisms may underpin future research into immune dysfunction and drug development aimed at alleviating severe and/or persistent disease in at risk patient populations.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY / ABSTRACT Many major life transitions, such as starting a new job or switching schools, involve learning new social structures. Successfully learning these structures is critically important for feeling a sense of belonging, improving academic or job performance, increasing well-being, and establishing relationships with others, but failure can increase feelings of social isolation and loneliness. These feelings are strongly associated with a greater risk for heart disease and other cardiovascular risk factors, such as hypertension and obesity. This striking relationship between loneliness and negative cardiovascular health outcomes is an urgent public health priority in the United States. While factors like social integration and adapting to novel social environments influence both mental and physical health, relatively little is known about exactly how people learn new social structures, and what factors make some individuals better social learners than others. Moreover, no empirical research to date has examined the relationship between social-relational learning and loneliness using a combination of behavioral, physiological, and neurocognitive methods. The proposed research aims to address these gaps by conducting a longitudinal study during the real-time formation of a social network and measuring biological and psychological signals that may explain individual differences in social learning success. In this study, 300 individuals from the first-year Honors program cohort of undergraduate students – who live and take smaller classes together – will complete a social-relational accuracy survey at three timepoints during the academic year. Each participant will receive a roster of names for all cohort members and will be asked to identify the strength of friendship with each name. Participants will additionally complete surveys assessing social integration, loneliness, and social support. We will test the hypotheses that more socially integrated and supported people will feel less lonely, and that social-relational accuracy is associated with individual differences in loneliness. A random subset of 60 individuals will additionally participate in a neuroimaging experiment using a previously-validated social-relational learning paradigm. Participants will view videos of interactions between contestants on a reality television show and rate the degree to which contestants are friends or rivals with each other. Resting heart rate variability (rHRV) will be measured continuously throughout the task. Using multivariate analysis techniques, we will identify patterns of activity for individuals who are better or worse at identifying the strength and valence of these viewed relationships. We will test the hypothesis that participants who are better at identifying the quality of these relationships will show increased neural synchrony in the dorsomedial and ventromedial prefrontal cortex, temporoparietal junction, and ventral striatum, will report feeling less lonely, show higher social-relational accuracy, and have a higher rHRV. By illuminating a variety of mechanisms that support social-relational learning and directly relating these mechanisms to individual outcomes, these findings may support the development of strategies to reduce loneliness and the risk of cardiovascular diseases.

NIH Research Projects · FY 2025 · 2024-09

Abstract Research has documented that interpersonal violence (IV; e.g. intimate partner violence, sexual assault, child abuse) disproportionately impacts sexual minority (SM) populations, with unaddressed victimization experiences representing a key driver of SM health disparities (e.g., suicidality, substance use, depression, HIV transmission, morbidity, mortality). To advance from identifying these disparities to intervening, research that evaluates SM barriers to seeking help and IV recovery needs, as well as the social, cultural, and structural influences of these processes at multiple levels is required. There is a scarcity of tools to measure IV-specific concepts validated in SM populations, with no current instruments to measure the help-seeking barriers SM survivors face and their recovery progress. Thus, there is a critical need to develop these instruments to inform acceptable and effective interventions to increase the access, engagement, care linkages, and effectiveness of survivor support infrastructure for SM populations. Through an exploratory sequential mixed-methods approach, we aim to: 1) develop an SM Barriers to Help-seeking and an SM IV Healing Scale and 2) psychometrically evaluate these instruments. Study aims will draw on four sequentially collected sources of data: a) an existing dataset of SM ethnographic narrative interviews (N=40), b) data gathered from our community advisory board (N=10), c) cognitive interviews with SM survivors (N=20), and d) a national online survey (N=1000), with adequate representation of sexual minority men and women. A community advisory board with SM IV survivors, clinicians, scientific experts, and community stakeholders will be engaged throughout the research process ensuring potential impact and sustainability. This study is innovative because it uses rich community-engaged approaches to articulate help-seeking and recovery experiences at multiple levels, incorporating SM survivors at all stages of the process, with an emphasis on recovery rather than deficit. In turn, the instruments created are expected to open new horizons in SM survivorship research and practice, specifically the ability to illuminate SM recovery needs and prioritize interventions to mitigate drivers of SM health disparities. Successful completion of this project is expected to have a positive impact by enhancing our ability to: 1) evaluate future help-seeking and recovery intervention effectiveness, 2) enhance help-seeking and recovery exploration in SM populations, and 3) determine how changes influence help-seeking and recovery for SM populations. The study's key deliverables will include two instruments measuring help-seeking barriers and IV recovery needs, validated in SM populations, using rigorous community-engaged qualitative, quantitative, and mixed methods approaches. These expected outcomes will support NIH priorities for Health Disparities Science by strengthening measurement of help-seeking barriers and recovery in SM survivors, thereby improving understanding on multi-level areas for additional exploration to improve health disparities in these populations.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Child sexual abuse (CSA) is an adverse childhood experience with wide ranging effects on health, wellbeing, and development throughout the lifespan. The long-term goal of this Stephen I. Katz, Early-Stage Investigator R01 is to develop novel, targeted prevention approaches for recurrent CSA, a new research direction for the PI (Kobulsky). The objectives are to illuminate the etiology of recurrent CSA and to conceptualize prevention models within child welfare systems. To accomplish this, we propose a mixed methods design to reduce recurrent CSA via existing partnerships among Temple University researchers, practitioners at the Philadelphia Children’s Alliance (PCA, Child Advocacy Center), and administrators in the Philadelphia Department of Human Services (DHS; jurisdictional CPS agency). Our team of experts on CSA prevention, data science, geographic spatial analysis, and qualitative and mixed methods positions us to create and analyze a unique linked longitudinal dataset, and to conceptualize prevention that will integrate research findings and the perspectives of multiple system constituents. Aim 1 is to curate a comprehensive longitudinal dataset from 2013-2021 including both CPS and law enforcement-investigated cases of CSA. We will leverage state-of-the-art data science techniques to create the dataset through the linkage of the PCA and DHS administrative data (Aim 1a). Natural language processing will be applied to extract data elements from free text fields, (Aim 1b) and geocoding to link neighborhood-level information. This unique dataset allows us to conduct a rigorous study of CSA and gain new insights into the prevalence and contributing risk and protective factors to CSA over time. Aim 2 is to illuminate the multilevel etiology of recurrent CSA. The aim will apply shared frailty survival analyses and spatial survival analysis to the dataset created in Aim 1 in order to identify the most potent predictors of recurrent CSA over time. It will examine the influences of CSA characteristics (Aim 2a), and social determinants of health (SDH) at the family (Aim 2b) and neighborhood (Aim 2c) levels on CSA recurrence. Aim 3 is to conceptualize prevention models for recurrent CSA. Qualitative interviews with multidisciplinary workers from the Philadelphia child welfare system, non-offending caregivers, and survivors will elicit interpretation of Aim 1 and 2 findings to enrich our theoretical model and conceptualize novel recurrent CSA prevention. Using a cutting-edge, mixed-method design, this project will illuminate the etiology of recurrent CSA and present significant new avenues for CSA prevention embedded within child welfare systems. This grant is an essential first step in a R34 NIH planning grant to develop novel, targeted CSA prevention, a new research agenda for the PI that can advance life course health and development for survivors of CSA.

NIH Research Projects · FY 2024 · 2024-09

SUMMARY Adverse childhood experiences (ACEs) can increase the risk for obesity both during childhood and later in adulthood. Emerging evidence suggests a parent's history of ACEs can increase their child's obesity risk, regardless of whether the child experiences their own ACEs. The association between a parent's ACEs and their child's obesity is believed to be moderated by via enduring stress-related effects of ACEs on parent's biology and behavior and family functioning. Research on ACEs-obesity associations rarely focuses on neighborhood environment, yet neighborhood characteristics are relevant to both ACEs and obesity risk. Neighborhood characteristics such as greenspace and food access can impact engagement in obesity related behaviors. Neighborhood factors such as neighborhood poverty and crime can be a source of adversity. Our previous research documented that adults with a history of ACEs live in less health-promoting neighborhoods with more obesogenic qualities (e.g., worse supermarket access, less greenspace). However neighborhood's impact on associations between a parent's ACEs and their child's obesity has not been widely investigated. Illuminating the role of neighborhood requires advanced spatial analytic methods, because neighborhoods are complex environments comprised of numerous synergistic characteristics that co-occur within a broader context of racial and economic segregation. To begin to illuminate neighborhood influences on ACEs-obesity associations, we validated spatial analytic methods for creating a Neighborhood ACEs Index (NAI). A NAI is a weighted composite that answers the question “what is the neighborhood environment of individuals who have experienced ACEs?” It serves as a single-item individual-level proxy for collective neighborhood environment associated with ACEs. Individuals who live in neighborhoods that have more characteristics associated with ACEs have a higher NAI score. Using geocoding and spatial methods, a NAI connects individual-level ACEs data with rich spatial data that includes numerous neighborhood characteristics such as crime, poverty, and healthcare access. A NAI can be used as an independent variable in analyses to examine how neighborhood characteristics associated with ACEs influence the relationship of ACEs and obesity. In addition, a NAI identifies population-specific neighborhood-level targets for interventions and polices. Neighborhoods whose residents have the highest NAI score can be prioritized for trauma-informed obesity reduction efforts. Building upon our prior work, this study will create a NAI based upon parent ACEs and test how that NAI influences associations between a parent's history of ACEs and their child's obesity. We will harness robust existing data from the H2O study (N=1320) that captured parent ACEs and child obesity, rich spatial data that captured neighborhood environment, and robust spatial methods. Collectively, this study is a foundational step towards our long-term goal: informing development of place-based interventions and policies to reduce obesity for millions of Americans affected by ACEs.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Oral squamous cell carcinoma (OSCC) exerts a significant clinical and financial burden worldwide. OSCC continues to have poor prognosis despite advances in treatment modalities including surgery, radiotherapy and immunotherapy, highlighting the urgent need for development of novel therapeutic strategies for this deadly disease. Recently, there has been increasing interest in the role of the microbiome in OSCC. Extensive research—including studies by Dr. Al-Hebshi—has shown OSCC to be associated with a distinct microbiome. However, mechanistic in vitro and animal studies have almost exclusively focused on exploring of the role of tumor-associated species (i.e. the pathobionts) while overlooking the potentially protective role health-associated species could play in oral carcinogenesis. Indeed, there is growing evidence to support the use of natural and engineered bacteria as anti-tumor agents. Additionally, studies have found the microbiome to modulate response to cancer immunotherapy. However, such studies in the context of oral microbiome and oral cancer are lacking. Our overarching goal is to identify oral bacterial species with anti-cancer properties and exploit them for prevention or/and treatment of OSCC. Our central hypothesis, supported by preliminary data, is that health- associated oral bacteria can interfere with OSCC tumor development and progression by inducing cytotoxicity in cancer cells, inhibiting proliferation, downregulating CD36, and/or modulating the microbiome. We also hypothesize that health-associated oral bacteria activate antitumor immunity, which in turn can enhance efficacy of immunotherapy. To address these hypotheses, we propose the following Specific Aims: Aim 1 To assess effect of S. mitis and H. parainfluenzae on development of chemically-induced OSCC in vivo (Prevention setting); Aim 2 To study the ability of these species to modify the response of chemically-induced OSCC to checkpoint inhibitor immunotherapy (Adjunctive therapy setting). We will employ a range of technologies including digital imaging, histopathology, immunohistochemistry, flow cytometry, q-PCR, fluorescent in-situ hybridization and 16S and RNA sequencing to investigate the effect of treating 4-NQO mice with the test species. The proposed studies explore for the first time the novel concept of using health-associated bacteria as anticancer and immunomodulatory agents in OSCC. These studies have high potential for translational impact in OSCC. Given the accessibility of the oral cavity, local delivery of bacterial species with anti-cancer properties via gel, mouthwash, or lozenge may be used in human subjects as a new paradigm for OSCC prevention and/or as adjunctive therapy. An additional strength of our approach is that it employs health-associated bacteria, that are unlikely to be cleared by the immune system or to induce adverse effects.

NIH Research Projects · FY 2025 · 2024-09

Cardiovascular diseases remain the leading cause of deaths globally, representing approximately 31% of deaths per year, 85% of which are due to myocardial infarction (MI) or stroke. This does not account for the millions of patients suffering daily with current therapies that are not efficacious in treating a major cause of cardiovascular disease: atherosclerosis. Current therapeutics for atherosclerosis fail to consider the role of dampening inflammation and improving lymphatic function, both of which have the potential to promote plaque regression. Lymphangiogenesis is the process where the lymphatic network is extended, but its role in atherogenesis is currently under-characterized and controversial. Inflammatory diseases are found to have lymphangiogenesis with associated malfunction, but controversy exists as to whether this is a cause of disease pathogenesis or a compensatory attempt at disease resolution. Recent data suggests a role for lymphatic vessels in reverse cholesterol transport (RCT), which relies on the junctional morphology between lymphatic endothelial cells (LECs) to regulate the egression of HDL and/or inflammatory cells. During inflammation, junctions between LECs transform from discontinuous buttons to impermeable zippers, which may impede RCT and prevent resolution of inflammation. Our lab has previously reported that Interleukin-19 (IL-19), an immuno-modulatory cytokine, uniquely attenuates atherosclerotic plaque progression while also being pro-angiogenic. This drives our central hypothesis that one mechanism whereby IL-19 prevents atherosclerotic disease progression is by driving functional lymphangiogenesis, especially through the maintenance of permeable lymphatic vessels to promote RCT. Our in vitro data indicates IL-19 can significantly induce primary LEC proliferation, migration, and tube formation, all lymphangiogenic assays. RNA sequencing also identified upregulation of Prox1, the master transcription factor of lymphangiogenesis, 6.2-fold. IL-19 also increases expression of several genes implicated in endothelial cell permeability, including Angpt2. Functional in vitro permeability assays showed that oxLDL decreases LEC permeability which IL-19 treatment prevents. This suggested that IL-19 can promote transport of lipid through lymphatic vessels and was confirmed with HDL transit. Junctional morphology was also validated via immunocytochemistry for VE-cadherin on hdLECs. Western blot confirmed phosphorylation of STAT3, a factor in canonical IL-19 signaling, and VE-cadherin, an indication of LEC permeability. Subsequent Prox1 siRNA knockdown mitigated both IL-19 stimulated lymphangiogenesis and Angpt2 upregulation. Preliminary in vivo data with VEcadCreERT2 Prox1 F/+ mice, which experience compromised lymphatic function, indicates that IL-19 improves lymphatic function through removal of Evans blue dye. These data suggest that IL-19 increases lymphatic vessel formation and function, with a potential mechanism of enhancing RCT to decrease plaque burden in atherosclerosis. This grant will continue to characterize IL-19’s mechanistic pathway in LECs and determine lymphatic functionality in vivo through HDL and foam cell trafficking by aortic microlymphangiography.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Chronic Respiratory Diseases (CRDs) are the third leading cause of death and disability in the United States, with a significant impact on the sociobiological and economic health of the country. Many patients with CRDs present with dysregulation of lung healing after injury. Therefore, there is an unmet need to better understand the molecular mechanisms underlying lung regeneration in order to develop novel treatment strategies for these patients. Most of the gas exchange region of the lung is lined by Alveolar Epithelial Type 1 (AT1) cells. When these cells are damaged, Alveolar Epithelial Type 2 (AT2) cells activate, proliferate, and differentiate into new AT1 cells to replace them. Failure of AT2-to-AT1 differentiation contributes to the pathogenesis of CRDs. However, the mechanisms underlying this transition are not completely understood. Data suggest that expression of Mitochondrial Calcium Uptake 1 (MICU1) protein, as well as a reduction of trimethylated histone H3 residues lysine 9 (H3K9me3) and lysine 36 (H3K36me3), are both vital for AT2-to-AT1 differentiation. We hypothesize that alterations in MICU1-dependent mitochondrial calcium uptake drive AT2 differentiation by rewiring cellular metabolism to favor the accumulation of α-ketoglutarate at the expense of succinate, thereby increasing the activity of α-ketoglutarate-dependent histone demethylases and promoting an AT1 pattern of gene expression. Therefore, our specific aims are to: (1) Determine the distribution of H3K9me3 and H3K36me3 marks across the genome, and their functional consequences, during AT2-to-AT1 cell differentiation in AT2 wildtype and MICU1 knockout cells, and (2) Determine whether increased H3K9me3 and H3K36me3 are responsible for the decreased AT2-to-AT1 differentiation capacity of AT2 MICU1 knockout cells. Aim 1 will utilize CUT&RUN, ATAC-Seq, and RNA-Seq, while Aim 2 use CRISPR/Cas9 gene editing both in vitro and in vivo. The proposed experiments will take place in the Center for Translational Medicine in the Lewis Katz School of Medicine at Temple University, an institution with a track record of excellence in both clinical medicine and scientific research. The training plan, developed by Co-Sponsors Drs. Tian and Elrod, will be tailored to ensure success of the applicant, Morgan Pantuck, as she progresses through Temple's MD/PhD program. Overall, this grant will provide an excellent training vehicle for the applicant while also increasing our understanding of the molecular mechanisms underlying lung regeneration, supporting development of novel treatments for CRDs.

NIH Research Projects · FY 2024 · 2024-09

Project Summary Craving, the intense desire for drugs, is a pivotal cause of relapse in substance use disorders and is known to grow during abstinence known as the incubation of craving. This phenomenon is prominent in both humans and animals yet is measured divergently—through instrumental behavior tasks in animals and subjective assessments in humans. This discrepancy underscores the differing cognitive factors driving craving where animal models focus on habitual behaviors driven by striatal learning systems whereas human models assess self-reported feelings associated with craving driven putatively by the hippocampus, a neural system associated with reward memory. The traditional understanding of reward learning, driven by the striatum, posits that memory for rewards should fade during abstinence, but the plethora of incubation data shows an escalation. Hippocampal memory systems provide a mechanism by which reward memories can intensify over time via systems consolidation processes which replay memories to transform their traces into cortical regions such as the prefrontal cortex (PFC), another region implicated in cue-induced craving. Novel methods are needed to relate trans-species methods of craving in a single paradigm capable of teasing apart cognitive-emotional mechanisms that may comprise different elements of craving. F99 Phase: The proposed pre-doctoral research will utilize a novel fMRI paradigm exploring craving in a normative human population characterizing the incubation of craving for non-drug rewards, i.e. foods, using subjective and instrumental assessments of craving. In this way, we can leverage idiosyncratic items that individuals have a history of naturalistic encounters and display a range of liking and times of abstinence since last consumption. Utilizing fMRI, we will examine the mesolimbic circuitry related to habitual elements of craving, hippocampal activation related to cue-induced craving, and prefrontal involvement in storing memories over varying lengths of time. Additionally, through natural language processing of reward memory narratives, we aim to construct linguistic profiles correlating subjective craving with behavioral expressions, broadening our tools to understand craving. This will enable a multifaceted understanding of craving, integrating instrumental and subjective dimensions. K00 Phase: This phase extends this integrated model to study craving I will have developed in graduate school to individuals with nicotine use disorders, investigating how craving memories differ across natural and drug rewards and their impact on craving intensity and duration. This approach aims to delineate the complex dynamics of craving in SUDs, contributing to the development of targeted interventions. This comprehensive exploration will advance our understanding of craving's neurobiological underpinnings, bridging the gap between subjective experiences and observable behaviors. The findings will have significant implications for developing effective treatments for addiction, aligning with the applicant's long-term objective of pioneering a research lab dedicated to the cognitive neuroscience of addiction and substance use disorders.

NIH Research Projects · FY 2025 · 2024-09

The intracellular pathogen Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis (TB), which has been one of the most deadly infectious diseases globally for decades. TB is not on target to be eradicated, and multi drug-resistant (MDR) strains of Mtb are a growing world health problem, with 500,000 new cases of MDR-TB reported annually. An additional challenge is the resistance of dormant mycobacteria to current antibiotic treatments, leading to long treatment regimens (ranging up to 2 years). It will thus be critical to build on our understanding of the molecular physiology of mycobacteria to gain insight toward improved anti TB treatments. A strategy for new drugs to kill dormant and MDR Mtb is to target Mtb ATP synthesis pathways, and a potential class of molecular targets includes K+ channels, which can modulate membrane potential to affect ATP synthase activity and ATP production. Insertion mutagenesis screens of Mtb have implicated the putative K+ channel-encoding gene Rv3200c as a potential virulence gene (i.e. critical for Mtb survival following infection), although functional, mechanistic studies of this gene have not been pursued in detail. In preliminary experiments using the non-pathogenic model organism Mycobacterium smegmatis, we have found that deletion of the Rv3200c ortholog (Msmeg_Δ1945) leads to decreased cellular ATP levels, consistent with a role for this channel in modulating membrane potential to support ATP production. In addition, our preliminary studies using K+ channel-deleted Mtb (Mtb_ΔRv3200c) further show that MycK deletion enhances sensitivity to killing by the H+/K+ ionophore nigericin (which decreases the transmembrane H+ gradient). These preliminary results support the working hypothesis that the Rv3200c-encoded K+ channel, termed MycK, may be a key modulator of the protonmotive force to regulate mycobacterial ATP levels. To better understand fundamental mechanisms underlying modulation of the MycK channel, as well as its role in mycobacterial physiology, we propose to 1) determine the ligand activation mechanism of the MycK channel by solving its structure by single particle cryo-electron microscopy, and by identifying the gating mechanism through analysis of single-channel recordings of purified MycK channels; and 2) determine the physiological role of MycK in control of energy metabolism, by measuring ATP levels and metabolic activity in gene-targeted mycobacteria. The new structural and functional data generated in this multi-disciplinary project will contribute to the ultimate goal of developing improved therapies for TB and other mycobacterial infection.

NIH Research Projects · FY 2024 · 2024-09

Supplement Summary and Aims The funded grant aims to determine the role of microRNAs (miRNAs) in post- transcriptional regulation in the context of cerebrovascular dysfunction and blood-brain- barrier (BBB) disruption, particularly within Alzheimer's disease (AD) and cerebral amyloid angiopathy (CAA). AD is characterized by an accumulation of amyloid-β (Aβ) plaques and neurofibrillary tangles, with cerebrovascular dysfunction (CVD) and neuroinflammation playing key roles in the progression of the disease. In fact, most cases of AD exhibit Aβ deposition in and around cerebral blood vessels, thus presenting an additive effect to the pathology of the disease. The BBB, composed mainly of endothelial cells (ECs), regulates the entry of circulating substances from the blood to the brain and assists in clearing harmful molecules from the brain, including oligomeric Aβ. Disruption of the BBB and the accumulation of Aβ in and around cerebral blood vessels is associated with changes in ECs, yielding a proinflammatory phenotype and leading to apoptosis. In our preliminary data of the parent grant, we have found that miRNA-212 is decreased in ECs treated with Aβ. Previously, we have shown that Aβ treatment leads to EC apoptosis and increase in EC barrier permeability. We have shown in our preliminary data that several genes involved in EC apoptosis contain binding sequences for miRNA-212. Our overarching goal is to test the hypothesis that Aβ promotes cerebrovascular dysfunction and BBB failure via endothelial cell miRNA-212. To achieve our goal, during this Supplement I specifically aim to accomplish the following Aims. Aim 1: measure miR-212 levels in exosomes released from ECs under the same Aβ conditions as in Aim 1 of the Grant. The release of miRNAs via exosomes may be essential for regulation in nearby cells of the neurovascular unit and may serve as a biomarker for AD and CAA diagnosis and progression. Aim 2: assess the impact of miR- 212 downregulation and rescue on BBB permeability in neurovascular unit complex systems. To ascertain the role of miRNA in CVD and BBB loss of function.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Currently, more than 6 million Americans are suffering from clinical Alzheimer's disease and Alzheimer's related dementias (AD/ADRD). By 2050, this number is projected to rise to 13 million. Due to this expected increase, there is a great need for sensitive, easily implemented, objective, and scalable methods to identify cognitive change and differentiate individuals along the AD continuum. Neuropsychological assessments are useful for characterizing dementia symptoms, clinical diagnosis, and monitoring, but they involve burdensome procedures and lack ecological validity. Neuroimaging and blood markers of proteins associated with risk of clinical AD/ADRD are often inaccessible, expensive and/or invasive, and their link to patient behavior is inconsistent. To address these drawbacks, digital phenotyping is a novel approach that allows for passive and continuous monitoring of behaviors and physiological metrics in everyday life using technology such as a smartphone or smartwatch. In this study, the construct validity of smartwatch-derived metrics will be investigated against conventional neuropsychological measures and questionnaires across preclinical AD/ADRD stages. A diverse sample of 90 participants aged 55 and older with either healthy cognition or mild cognitive impairment will be recruited from the Philadelphia region. Participants will be given a Garmin Vivosmart 4 smartwatch and will wear it 23 hours/day to passively monitor sleep, heart rate variability, and physical activity data, which then will be deidentified and transferred to a secure server daily for 30 days. A daily survey will be completed through study software that is downloaded on participants' smartphones to contextualize passive data collection metrics. At the first study visit, participants will complete conventional neuropsychological tests and self-report measures of everyday function. The primary aim will examine the validity of a smartwatch-derived digital phenotyping protocol against conventional clinical measures. Exploratory aims will investigate 1) associations between smartwatch metrics and demographic and contextual factors and 2) meaningful groupings of participants with unique patterns of smartwatch metrics (i.e., phenotypes) and their relations to clinical status and conventional cognitive measures. The proposed training plan was developed with input from a team of interdisciplinary experts in everyday cognition in aging, digital ethics, longitudinal statistical methods, and engineers studying physiology with the goal of developing a high level of competence in statistical methods for wearable devices, understanding physiological measurement, knowledge on ethics and privacy in the wearable space, and the application of digital phenotyping in a diverse aging population.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Childhood apraxia of speech (CAS) is a pediatric speech sound disorder (SSD) that impacts between 60,000 and 780,000 children under 14 in the US. The underlying impairment in speech motor planning primarily affects articulation and prosody and reduces intelligibility. Beyond impaired communication, children diagnosed with CAS are also at higher risk for difficulties with social/emotional development and reading and writing. Thus, effective targeted intervention is warranted for these children. However, CAS diagnosis is difficult and at present relies primarily on the `gold standard' clinical judgment by an `expert speech-language pathologist (SLP)' based on a number of perceptual features: (a) Inconsistent errors, (b) impaired transitions between sounds and syllables, and (c) impaired prosody, in particular lexical stress. This diagnostic standard is problematic for several reasons, including the fact that these features are not operationalized (in terms of elicitation, measures, number of instances required for diagnosis), lack precision, and vary with the SLP's language and clinical experience. These challenges are exacerbated by the well-known heterogeneity of CAS presentation and the overlap with other SSDs. Finally, there are no standards or criteria by which an SLP is considered an `expert'. Together, these major limitations lead to disagreements between SLPs and reluctance by some SLPs, especially less experienced ones, to diagnose CAS. This undermines the ability to make well-informed evidence-based clinical decisions for children with CAS and other SSDs. In addition, the research literature on CAS has been hampered for decades by this diagnostic problem, and is compounded by small sample sizes. More objective, validated, precise, replicable, and clinically feasible diagnostic measures are sorely needed. Acoustic measures have strong potential to meet these needs, based on prior small-scale studies in CAS and studies with other populations with speech disorders. However, to date no studies have systematically examined acoustic measures of core perceptual CAS features in a single study. This F31 proposal leverages a unique, large and well-characterized retrospective dataset of 125 children (76 with CAS, 28 with another SSD, and 21 without any SSD) to systematically examine the validity of a well- motivated set of acoustic measures of each core perceptual feature (3 acoustic measures each) (Aim 1), the differential diagnostic accuracy of the three acoustic measures with highest validity (1 acoustic measure per feature) (Aim 2), and the potential identification of CAS subtypes based on these acoustic measures (Aim 3). As a first step, the diagnostic accuracy of these acoustic measures will be evaluated against a gold standard of an operationalized consensus criterion based on independent judgments from 3 expert SLPs. This F31 project will provide novel, unique, and significant information that will help improve clinical diagnosis of CAS and other SSDs, provide the Fellow with advanced training to transition toward an independent research career, and set the stage for a future prospective validation study of the acoustic measures found most promising here.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY Oral pathogenic biofilms are major drivers of major oral infectious diseases such as dental caries and denture stomatitis. Currently, the annual cost of treatment of oral biofilm-related infectious diseases exceeds $81 billion only in the US. Strategies to control biofilm formation on dental biomaterials are needed. Biofilm formation and adhesion are complex processes controlled by the interplay between physicochemical and biological processes including microbial strain, environmental conditions, and biomaterial surface. Specifically, biomaterial-biofilm interactions are controlled by different biomaterial surface properties including wettability, charge density, stiffness, roughness, topography, and chemistry. Biomaterials in the oral cavity are subjected to cyclic loading from masticatory and biomechanical movements. These repetitive forces can have an effect in the biomaterial-biofilm interactions. In fact, our recent publications and preliminary results, we showed for the first time, that the cyclic strain of the biomaterial surfaces is a parameter influencing the virulence and dysbiosis of oral microbes (Candida albicans and multispecies biofilms), which can lead to the progression of infection and disease such as denture stomatitis. The overarching goal of this proposal is to mechanistically study the effect of biomaterials’ cyclic strain on the pathogenesis of oral microbes. In this study, we will focus on the pathogenesis a fungal strain (Candida albicans) and denture-microbiota. Aim 1 will study this effect using common materials used in dentures. Aim 2 will explore the phenomena with hard oral mucosa and Aim 3 with an animal model. This project will help understand how the action of chewing is fueling the progression of major oral infections such as denture stomatitis, which in turn could lead to the development of methods for the prevention of pathogenic biofilm formation over implanted medical devices.