University Of Miami School Of Medicine

NIH Research Projects · FY 2025 · 2024-07

Aging and HIV are characterized by systemic chronic inflammation (inflammaging) and chronic immune activation (IA), both associated with cell senescence, functional impairment of immune cells and reduced immunity to infections and vaccines. Aging and HIV are also associated with metabolic changes such as decreased insulin sensitivity, dysregulated nutrient uptake and dysfunctional mitochondria. As a consequence of these changes, metabolites involved in glucose and fatty acid metabolism are released in blood. Some of these metabolites have been shown to be positively associated with inflammaging and therefore expected to be also associated with dysfunctional immunity. In people living with HIV (PWH) metabolic abnormalities have been related to virus-induced irreversible tissue damage in viremic individuals, as well as to anti-retroviral therapy (ART)-induced effects in virally suppressed individuals. Immune cells need to undergo metabolic reprogramming to meet the demands associated with immune responses, and rely on anaerobic glycolysis and oxidative phosphorylation to do so. The effects of aging in PWH on these pathways and on the metabolic mechanisms involved in the regulation of immune cell function are largely unknown and are the primary objectives of this proposal. Our hypothesis, supported by strong preliminary results, is that aging exacerbates immune defects through a dysregulated metabolic reprogramming and this effect is even more pronounced in PWH. Our studies have demonstrated that a higher metabolic status of immune cells is associated with higher intrinsic IA and lower capacity to generate protective immunity. This proposal will use data and samples from the MACS/WIHS Combined Cohort Study (MWCCS) of PWH and age- gender- and sociodemographically matched people without HIV (PWoH). We plan to evaluate how aging influences the serum metabolic profile of PWH and how this impacts the function, phenotype and transcriptional profiles of B cells, T cells and monocytes, the major cell types participating in immune responses to infections and vaccines. The role played by gut microbial metabolism on metabolic status will also be evaluated (Aim 1); we will investigate the metabolic mechanisms as well as metabolic requirements of the immune cells (Aim 2); and we will perform experiments using senolytics and metabolic modifiers to block senescent and metabolic pathways and improve, at least in vitro, immune cell function (Aim 3). Because obesity is a condition associated with altered metabolism, experiments will use samples from lean and obese PWH of different ages, as evaluated by body composition measures such as weight, waist/hip/thigh circumference, and bioelectrical impedance analysis to assess body fat amounts and depots. Younger (<50 years) and older (>60 years) lean and obese PWoH will also be included as controls. Results obtained will allow the identification of senescent and metabolic pathways to be targeted to improve the immune function of PWH up to the levels observed in PWoH controls.

NIH Research Projects · FY 2025 · 2024-07

The All of Us Research Program (AOURP) is a historic effort to enable 21st century biomedical research that will benefit all persons in the USA. This study will power genetic studies of health and well-being, which will eventually deliver more precise treatments and personalized medications. A specific strength of the AOURP is the role of Regional Medical Centers (RMC/HPO), which provide their experienced staff and faculty, their resources, and their trusted relationships with patients and communities, to engage, enroll, and retain participants for the AOURP study. We have formed a consortium between the University of Miami and the University of Puerto Rico Comprehensive Cancer Center. Both institutions combined have enrolled close to 20,000 participants into AURP in the prior finding cycles. In this new award we will focus on engagement with existing participants and inform them of the continued opportunities the AOURP program has to offer. The University of Miami site will also enroll 825 participants with prior or continued opioid use disorder – this will happen with the joint support of NIDA. This study is thus part of the NIH’s Helping to End Addiction Long-term (HEAL) initiative to bolsters research across NIH to address the national opioid public health crisis and improve treatment for opioid misuse and addiction. Our experienced trained staff will use multiple measures of communication to keep participants engaged. Enrollment of participants will happen in our specialized AOURP sites and the IDEA clinic at U of Miami. With over six years of experience and a well-trained and motivated team, SEEC-2 will continue our successful engagement and enrollment strategies in the coming years.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY/ABSTRACT The goal of this F31-Diversity application, submitted by a predoctoral investigator from a disadvantaged background, is to examine the configurations of the online gaming and offline friendship networks of Latinx men who have sex with men (LMSM) and the extent to which these network structures and characteristics can influence LMSM access to pre-exposure prophylaxis (PrEP) information and the distribution of HIV self-testing (HIVST) kits. Despite the field’s infancy, online game-based interventions have been found to be acceptable among sexual and gender minority adolescents and adults and have positive effects for increasing knowledge, and improving attitudes and behaviors related to HIV prevention. Few studies have investigated the combined use of online game-based and offline friendship network approaches to increase awareness and uptake of PrEP/HIVST in the LMSM community. This mixed-methods social network study will analyze and integrate two data sources collected specifically for this F31-Diversity project as part of Dr. Mariano Kanamori’s (Main Sponsor) R01 entitled, “PrEParados: A Social Network Study of Latino MSM for Facilitating Progress in the PrEP Cascade” from April 2022 to August 2023 (R01MH12572). First data source: egocentric and two-mode network data (N=73 egos and 153 alters). Second data source: qualitative data including 90-minute semi-structured individual interviews with LMSM online gamers (N=40). Both data sources include information from LMSM living in Miami- Dade County, Florida, (MDC), the metropolitan statistical area with the highest HIV incidence (42.4 per 100,000) and second highest HIV prevalence (979.9 per 100,000) rates in the nation. Among incident HIV cases in MDC, 64.4% are attributed to Latinx individuals, and 81.5% are attributed to MSM. Design. The Network Flow Model and Social Contagion Theory will guide the following research aims: Aim 1: Characterize social networks of LMSM online gamers. It will include the description of network size, type of games played, frequency of gameplay with others, format of gameplay, and the sociodemographic characteristics of their online gaming partners. Aim 2: Understand the potential reach of LMSM online gamers to promote PrEP messaging and disseminate HIVST kits in their offline friendship networks. Egocentric and two-mode network analyses will be used to: (A) determine the structures of offline friendship and affiliation networks of LMSM online gamers, and (B) identify specific characteristics of these structures (e.g., type of games played, homogeneity, homophily) associated with the dissemination of PrEP information and distribution of HIVST kits. Aim 3: Explore the acceptability of a combined online game and offline friendship networks-based approach to promote PrEP information and HIVST kit dissemination. Qualitative findings will inform the design of an intervention that bundles social network and online game-based approaches to increase PrEP/HIVST awareness and uptake. Implications. This F31 study will inform the development and implementation of culturally-tailored game- and social network-based interventions to enhance HIV prevention services for LMSM and the applicant’s future K99/R00 award development.

NIH Research Projects · FY 2026 · 2024-07

The National HIV Strategy encourages expansion of HIV testing to community health settings, emphasizing it is critical that all people, particularly groups like people who inject drugs (PWID), know their HIV status for rapid initiation of HIV treatment or potential initiation of PrEP. Despite recommendations from CDC that PWID be tested for HIV every year, only 55% report receiving an HIV test in the previous 12 months. Injection drug use (IDU) continues to be a leading cause of HIV infection with 11% of new HIV infections and 66% acute HCV infections attributable to IDU in 2020. In recent years, there has been a national increase in IDU-associated HIV and hepatitis C (HCV) outbreaks, driven primarily by the ongoing overdose crisis, creating an obstacle to meeting Ending the HIV Epidemic (EHE) goals. While the majority (87%) of syringe services programs (SSPs) offer HIV testing, it is estimated that participant-level uptake is only 17%, highlighting a significant practice gap regarding how and under what conditions SSPs are offering HIV testing. SSPs face a myriad of organizational-level barriers to implementing HIV testing, such as lack of financial resources, lack of training on best practices, staff turnover, and organizational capacity/policies. Our team developed, refined, and pilot tested the ACCESS implementation strategy—a multicomponent, multi-phase, organizational-level intervention focused on improving the SSP organizational capacity to implement HIV/HCV testing best practices through funding and practice facilitation. We propose a hybrid Type II effectiveness-implementation cluster randomized trial of 32 SSPs. We will create a cross-sector implementation team to facilitate the ACCESS implementation strategy in EHE jurisdictions and locales at risk for HIV outbreaks. Selected SSPs will then be randomized to receive the ACCESS strategy (i.e., funding and peer external facilitation), or an information control condition that contains a link to the CDC Compendium website and information regarding nationally available resources to support implementation. The primary outcome of the study will be HIV/HCV testing reach during the intervention period (12 months) compared to control sites. The aims of the study are: 1) to evaluate the effectiveness of the ACCESS strategy on participant-level uptake of HIV/HCV testing across 12 months; 2) to examine the effectiveness of our ACCESS implementation strategy on site-level adoption of opt-out testing and implementation fidelity; 3) to assess the cost-effectiveness of the ACCESS strategy in increasing uptake of HIV/HCV testing; and 4) to examine the sustainability of opt-out HIV/HCV testing at SSPs across 24 month follow-up. The cost-effectiveness analysis and mixed-methods implementation evaluation will provide compelling data on the impact of our ACCESS strategy on scaling-up routine opt-out HIV/HCV testing at SSPs. Leveraging the critical role of SSPs, the ACCESS strategy has the potential to make EHE an attainable reality for PWID.

NIH Research Projects · FY 2026 · 2024-06

Chronic infections and cancer cause CD8 T cell exhaustion, a differentiation state characterized by the inability of CD8 T cells to provide immunological protection. Improving the function of exhausted T cells (TEX) by antagonizing checkpoint inhibitors is a potent immunotherapeutic strategy. However, most patients do not exhibit long-term protection, creating a need for improved immunotherapeutic strategies. Resistance to immunotherapy is mainly attributed to the inability of checkpoint inhibitors to reverse the TEX epigenome. Therefore, epigenetically re-directing TEX differentiation to induce durable responses is key to overcoming resistance to immunotherapy. To date, modifying a single transcriptional pathway has had limited success in providing full TEX reinvigoration and inducing long-term persisting immune responses. Instead, microRNAs (miRs) can have a profound impact on the epigenetic regulation of immune cells; however, the role of miRs in TEX differentiation is understudied. We recently identified miR-29a as a unique memory-associated miR that is repressed in TEX. We demonstrated that ectopic expression of miR-29a enhances TEX persistence, attenuates their exhaustion, and promotes differentiation of a TEX subset with progenitor, stem-like characteristics that responds to immunotherapy. Importantly, our preliminary data suggest that miR-29a alters the epigenetic profile of TEX and promotes durable CD8 T cell responses, in striking contrast to anti-PD-L1 therapy. Mechanistically, we demonstrate that miR-29a directly targets the key DNA demethylating enzyme TET2 and that TET2 overexpression abrogates key effects of miR-29a in TEX differentiation. However, the molecular circuits involving miR-29a and TET2 and the implication of miR-29a-TET2 axis in checkpoint inhibitor therapy is unknown. The goal of this proposal is to uncover the role of miR-29a in targeting the key epigenetic modulator TET2 and, thus, define the role of miR-29a in synergizing with checkpoint inhibitors. Our central hypothesis is that miR-29a regulates DNA methylation by directly targeting TET2, therefore, synergizes with checkpoint inhibitors to re-direct TEX differentiation. Aim 1. Identify the role of the miR-29a-TET2 axis in regulating TEX differentiation. We hypothesize that miR- 29a regulates DNA demethylation, by directly targeting TET2, thereby fundamentally alters TEX differentiation. We will elucidate the TET2-dependent versus TET2-independent effects of miR-29a in TEX differentiation and the downstream effects in DNA methylation. Aim 2. Identify the pathways by which miR-29a synergizes with anti-PD-L1. We hypothesize that miR-29a epigenetically alters the differentiation of the progenitor TEX subset and, thereby, synergizes with anti-PD-L1 to promote durable TEX reinvigoration. We will elucidate the molecular mechanisms of synergy between miR-29a and anti-PD-L1 and we will define the TET2-dependent and TET2-independent roles of miR-29a in synergizing with anti-PD-L1 in mouse models of LCMV and B16 melanoma.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Obesity, physical inactivity, and poor diet are major modifiable risk factors for cancer — a leading cause of death among US Hispanics. US Hispanic youth have the highest rates of pediatric obesity and show faster increases in body mass index (BMI) from age 12 to <20 years than non-Hispanic Whites. Most Hispanic adults also carry excess weight, and Hispanic youth and adults do not often meet national guidelines for physical activity and dietary intake. Family-based interventions can improve health outcomes in both youth and parents and are highly relevant to Hispanics due to the cultural value of familismo (familism). However, very few existing family-based obesity prevention interventions for Hispanics target adolescents and their parents, and those that do are not designed to facilitate widespread reach. An underutilized approach for promoting a healthy weight status among Hispanics is through digital (i.e., eHealth/mHealth) interventions. US Hispanics are just as likely as non-Hispanic Whites to own a smartphone and more likely to be smartphone-only internet users. Our team employed a user- centered approach to develop and pilot a family-based digital lifestyle intervention known as Healthy Juntos (Healthy Together). This innovative six-month intervention (three-month intensive phase; three-month maintenance phase), grounded in family systems and behavioral theories, is one of the few obesity prevention interventions for Hispanic adolescents with digital delivery. It targets improvements in parent BMI and lifestyle behaviors, family functioning, and health-related positive parenting practices as the means through which adolescent BMI and lifestyle changes are hypothesized to occur. Unlike existing digital interventions for adolescents, the intervention makes parents the primary agents of change, with parent-adolescent participation a team effort through shared decision-making. Given promising findings for Healthy Juntos, the primary goal of this study is to evaluate, in a randomized controlled trial with 375 Hispanic adolescents and their parents, the efficacy of Healthy Juntos compared to a digital health referral to publicly available websites and apps (control) condition in preventing increases in BMI and improving moderate-to-vigorous physical activity (MVPA) and diet quality among adolescents. The aims of the study are: AIM 1: To determine whether participation in Healthy Juntos prevents increases in BMI (primary outcome) and improves MVPA, diet quality, and percentage body fat (%BF; secondary outcomes) in Hispanic adolescents. AIM 2: To examine whether and to what extent changes in parent BMI and lifestyle behaviors mediate the effects of Healthy Juntos on adolescent outcomes. AIM 3: To examine whether and to what extent family functioning and health-related positive parenting practices mediate the effects of Healthy Juntos on adolescent outcomes. We will also explore the role of sociodemographic (e.g., acculturation, adolescent sex, weight status, neighborhood deprivation) and user engagement as moderators as well as the mediating effects of individual-level constructs (e.g., self-efficacy, autonomous motivation).

NIH Research Projects · FY 2024 · 2024-06

Abstract Interstitial cystitis/ bladder pain syndrome (IC/BPS) is one of the most debilitating chronic pelvic pain (CPP) conditions that negatively impacts the quality of life and sexual activities in 2.7% to 6.5% of women in the US. Pelvic floor hypertonicity (PFH) is prevalent among patients with CPP, and is present in up to 85% of women with IC/BPS. PFH is a multifactorial problem that involves impaired relaxation of the pelvic musculature due to the activation of nociceptors and release of inflammatory chemicals. Current clinical strategies employ physical therapy interventions to relax the muscle to improve muscle coordination. Alternative therapies that use oral or intra-vaginal muscle relaxants may be suggested when conservative methods fail. Specifically, botulinum neurotoxin (BoNT), has received growing interest in managing pelvic pain secondary to PFH. BoNT inhibits the neuromuscular transmission at neuromuscular junctions, indicated by innervation zones (IZs), by blocking the acetylcholine release, leading to transient muscle relaxation, and the relief of pain symptoms. Despite its potency and safety, BoNT can cause dose-dependent adverse effects and is expensive. Studies have demonstrated that increasing the injection distance by 1 cm from the IZs reduced the effect of BoNT by 46%. It is clinically important to localize in vivo the IZ distributions in the pelvic floor muscles to guide precision BoNT injections for optimal treatment efficacy with reduced treatment cost. Our team has successfully developed a novel high-density surface electromyography (HD-sEMG) technique to 1) reliably and quantitatively assess PFH and 2) accurately image the IZ distributions of hypertonic PFMs, and has 3) successfully demonstrated the significant improvement in treatment outcome of BoNT injection with HD-sEMG vs. without HD-sEMG guidance, in managing muscle spasticity. The goal of this proposal is to develop a personalized, precision BoNT injection approach guided by intra-vaginal HD-sEMG to optimize the treatment outcome in IC/BPS management. This research represents the first effort to quantitatively and objectively study the feasibility of using intra-vaginal HD-sEMG for personalized, precision guided BoNT injections to optimize the diagnosis and treatment of IC/BPS. The proposed technique will serve as an in vivo pelvic floor phenotyping tool that can be used to deliver personalized injection of BoNT to personalized injection sites with personalized doses, which will lead to more effective BoNT treatment with reduced treatment cost (by reducing injection dose). The study will also advance our understanding of the pathophysiology of PFH, and mechanism of BoNT management, and help establishing an advanced standard of objective assessment of PFH and BoNT injections.

NIH Research Projects · FY 2025 · 2024-06

ABSTRACT Lung cancer is the leading cause of cancer-related deaths in the U.S., with ~236,700 new cases and ~130,200 deaths estimated for 2022. A joint effort by both PIs in this grant led to the identification of the oncogenic kinase protein kinase C epsilon (PKCe) as a key player in lung carcinogenesis. PKCe is aberrantly up-regulated in lung adenocarcinoma and is associated with poor outcome in patients specifically harboring KRAS mutations. Using genetically engineered mouse (GEM) models, we demonstrated that PKCe is required for both carcinogen- and Kras-driven lung tumorigenesis. In addition to this role in cancer initiation, our work established novel roles for PKCe in cellular events associated with late cancer stages. Indeed PKCe is a major player in lung cancer cell motility and invasion via activation of the small G-protein Rac1, linking this kinase to metastatic dissemination. That PKCe plays critical permissive roles first during adenoma initiation and then again later during the acquisition of metastatic competency highlights the functional complexities of the PKCe signaling events in lung cancer. Interestingly, CRISPR-mediated inactivation of PKCe in the initiating cell-of-origin does not significantly affect Kras- G12D-induced tumor development, leading us to hypothesize that PKCe does not strictly act in a tumor cell autonomous manner to permit oncogenic KRAS-mediated tumorigenesis. To test this hypothesis, in Aim 1 we will generate and characterize a series of GEM models to restrict genetic deletion of PKCe to either oncogenic Kras- expressing cancer cells or to diverse stromal cell types present in the tumor microenvironment, including non- cancerous epithelial, mesenchyme and hematopoietic cells. Gene expression studies on isolated cells using fluorescence-based lineage tracing techniques will provide significant mechanistic insights. In Aim 2, we will thoroughly dissect the mechanistic basis of motility/invasive signaling activation by PKCe. Our hypothesis is that PKCe activates Rac1-mediated formation of cell ruffles and motility in KRAS mutant lung cancer cells via Rac Guanine nucleotide Exchange Factors (Rac-GEFs). We will identify and characterize candidate Rac-GEFs as PKCe effectors responsible for this phenotype. In Aim 3, we will establish the involvement of PKCe and its effector Rac-GEFs for the development of metastatic lung cancer. We will use combined in vitro and in vivo approaches to pin down mechanistic defects in the metastatic cascade upon CRISPR-mediated deletion of PKCe and Rac-GEFs in lung adenocarcinoma cells. We will establish GEM models and use lentiviral CRISPR-based approaches to determine the permissive contribution of PKCe and its GEF effectors to lung adenocarcinoma metastasis, and identify relevant gene expression and signaling signatures contributing to this phenotype. Thus, by using innovative, state-of-the-art genetic and molecular approaches, our studies should reveal the vast multidimensional complexity of PKCe signaling in lung cancer development as well as underscore unappreciated mechanisms of carcinogen- and KRAS-mediated lung oncogenesis.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Infectious keratitis, also known as corneal infection, is frequently caused by bacteria, fungi, viruses, or parasites. Each pathogen needs a unique treatment approach. Hence, rapid determination of the underlying pathogen causing the disease is of utmost importance. However, current gold standard still includes culture of corneal samples, which not only can take days to identify the pathogen, but also still leads to false negative results in many cases. The delay in identifying the appropriate treatment strategy causes significant complications and unfavorable clinical outcomes, including blindness, which is why infectious keratitis is the 5th leading cause of vision impairment and blindness and the main cause of corneal blindness worldwide. Together, more rapid methods to determine pathogens are urgently needed, and would present a significant improvement in patient care. Here, we describe the development of a novel TaqMan-based assay, termed TLLPS (Thermal Lysis Low Input Pathogen Screen) that can rapidly distinguish between the most common pathogens causing infectious keratitis, with several advantages over other approaches: First, we designed our system to work with genomic DNA, which allows avoiding RNA-extraction, reverse transcription steps, and the significant challenge of heterogeneity emerging from isolating small amounts of heavily degraded RNA from the eye surface. Second, our assay allows using 1 µl of raw and un-processed biological fluids, which are submitted directly into a thermal lysis and genomic pre-amplification step, avoiding any purification steps, which would introduce variability and cause sample loss of the small amounts extracted from the ocular surface. Here we propose to expand and prospectively validate the assay. Further, we will test the use of ocular swabs for more rapid detection. With the completion of this proposal, we envision to have developed a clinically feasible, quick, and reliable way to identify the most common pathogens causing keratitis, and significantly improve patient outcomes through rapid identification of the correct treatment strategies. 1

NIH Research Projects · FY 2025 · 2024-06

There are more than 6 million cases of Alzheimer’s disease (AD) reported in the US, of which 10% live in Florida, making Florida the second leading state with the highest prevalence of AD. This number will rapidly grow, creating an urgent need to address this AD/ADRD public health crisis. This crisis is further exacerbated by a massive shortage of trained physicians and scientists caring for this vulnerable population. The proposed T32 program: Cross-disciplinary Research Opportunity for Training in AD/ADRD Science (CrossROADS) is a 2-year program at the University of Miami (UM), leveraging our experience implementing academic training programs over the past 20 years. The overall goal is to increase the number of AD/ADRD investigators-leaders in cross-disciplinary clinical and translational research, who can rapidly translate, implement and disseminate discoveries to practice and community. CrossROADS graduates will be capable to address the health challenges of AD/ADRD patients and caregivers we serve through team science and collaborations in partnerships with AD/ADRD community partners and stakeholders. CrossROADS is designed to select outstanding clinical, biomedical, and psychosocial science postdoctoral trainees, and provide a high-quality, research and leadership training program for the next generation of AD/ADRD scientists. The Objectives are to: (1) Develop and administer a 2-year cross-disciplinary research training program in AD/ADRD for 12 postdoctoral candidates; (2) Provide an evidence-informed mentorship through collaborative and individually tailored cross-disciplinary mentorship. Each trainee will be assigned to a mentorship team comprised of an experienced Primary Mentor, Peer Mentor, and as required, additional clinical or career development mentors to facilitate trainees’ socialization and integration in the academic cross-disciplinary research environment; and (3) Create opportunities to transition to career independence through career advising, coaching and sponsorship, strategies to enhance career visibility and competitiveness, and professional development with leadership training. We will integrate our trainees in our successful institutional training programs and collaborative environment fostering team science. The Leadership Team will rigorously evaluate the impact of the CrossROADS program on training of independent cross-disciplinary scientists-leaders, their mastery of core competencies, and their impact on health in AD/ADRD communities.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY Lipid-induced podocyte injury is an emerging molecular pathway contributing to the progression of glomerular diseases (GDs) of metabolic and non-metabolic origin. Research by others and us has highlighted a role for impaired reverse cholesterol transport (RCT) and altered sphingolipid metabolism in lipid-induced podocyte injury in GDs, yet, a common druggable pathway regulating both RCT and sphingolipid metabolism in podocytes remains to be identified. Among several lipoproteins, Apolipoprotein M (APOM) is mainly located in high density lipoprotein (HDL) particles where it facilitates RCT to HDL but also acts as a chaperone to transport sphingosine-1-phosphate (S1P) through the circulation. S1P signaling occurs through binding of APOM/S1P complexes to S1P receptors (S1PR1-5), which regulate many physiological processes, including migration, proliferation, and cell survival. We recently demonstrated significantly decreased glomerular APOM (gAPOM) mRNA expression in patients with GD enrolled in the NEPTUNE cohort. Our new preliminary data show that decreased gAPOM correlates with decreased plasma APOM (pAPOM), with increased glomerular sphingosine kinase 1 (SPHK1), the enzyme that converts sphingosine to S1P, and S1PR4 expression and with eGFR decline. We observed a similar gene expression pattern in Col4a3 KO mice, a mouse model of GD, and in Col4a3 KO podocytes, which was associated with glomerular/podocyte cholesterol and S1P accumulation due to impaired RCT and activation of S1P/S1PR4 signaling leading to increased apoptosis which was prevented by recombinant human APOM (rhAPOM) treatment. Importantly, the therapeutic effect of rhAPOM in preventing podocyte apoptosis in Col4a3 KO podocytes was superior to SPHK1 or S1PR4 antagonism. Finally, we demonstrate that treatment of human podocytes with exogenous S1P increases podocyte apoptosis and causes albumin leakage in a microfluid device as well as in ApoM deficient Col4a3 KO mice. Based on these observations, we hypothesize that GDs represent a state of gAPOM deficiency causing impaired RCT and activation S1P/S1PR signaling in podocytes, thereby causing lipotoxic podocyte injury. We propose a highly translational approach with three specific aims to 1) investigate if gAPOM deficiency correlates with the activation of glomerular S1P/S1PR signaling, is associated with decreased pAPOM levels and predicts outcomes in patients with GD, 2) to investigate the role of podocyte APOM deficiency on RCT and S1P/S1PR4 signaling, and 3) to investigate the therapeutic potential of recombinant human APOM in an experimental model of GD. If successful, this translational study may lead to the clinical development of APOM as a biomarker in GDs and to the use of recombinant APOM as a novel therapy for GDs.

NIH Research Projects · FY 2025 · 2024-05

Abstract/Project Summary Emerging preclinical data and clinical evidence support the existence of a bidirectional communication between the gut microbiome and the central nervous system. Recently, the gut microbiome has been shown to play a critical role in the development and function of the central nervous system (CNS). While advanced age as well as immune and metabolic alterations can lead to increased permeability of tissue barriers, evidence shows gut microbes signal to the brain even in young, healthy humans and in animal models. Signaling through the vagal nerve may play a role in these situations, but the exact mechanism of this communication remains to be confirmed. Bacterial extracellular vesicles (BEVs) have recently been shown to carry information over long- distances modulating interkingdom communication within the gut microbiota brain axis. Both gram negative and more recently gram-positive bacteria have been shown to release extracellular vesicles. These vesicles have been shown to contain a variety of biomolecules, such as proteins, lipids, nucleic acids, and small-molecule metabolites and play a key role in the biology of bacteria. BEVs has been postulated to be the key underlying mechanisms behind harmful or beneficial effects of many pathogenic, symbiont, and probiotic bacteria. Opioid use disorders have been associated with dramatic alteration in the gut microbiome with the expansion of gram- positive bacteria. Similarly, in humans and in animal models HIV infection is associated with microbial changes resulting in elevated systemic endotoxin levels-a signature marker for HIV co-morbidity. Opioid abuse exacerbates these effects both in PLWH and in animal models of HIV and substance use. The role of bacterial extracellular vesicles in modulating the gut brain axis under these conditions has not been investigated. This high risk, high reward R21 proposal is a pilot grant to establish proof of concept. Based on preliminary data and published studies, we hypothesize that microbial dysbiosis associated with opioid use and in the context of HIV, results in the expansion of distinct pathogenic bacterial communities leading to the secretion of BEVs which acts as mediators that communicates with both immune cells and microglial cells to disrupt the gut-brain-axis. Two specific aims are proposed to test this novel hypothesis. Specific Aim 1: Determine if bacterial EV’s derived from fecal samples from Control, Morphine dependent, HIV/ART and Morphine dependent/HIV/ART treated mice a) disrupt gut epithelial and brain endothelial barriers and b) cross gut epithelial and blood brain barriers using in vitro gut epithelial and blood brain barrier models. We will further investigate if BEVs modulate macrophage and microglial gene expression in co-cultures. Specific Aim 2: Determine if Extracellular vesicles isolated from fecal samples from Control, Morphine dependent, HIV/ART and Morphine dependent/HIV/ART treated mice a) disrupt epithelial barrier b) increase gut epithelial permeability and c) cross gut epithelial barrier and Blood brain barrier in vivo. The clinical relevance of these studies is that BEVs can be exploited for therapeutic purposes to deliver nano pharmaceuticals systemically and to the CNS.

NIH Research Projects · FY 2026 · 2024-05

Project Summary Per- and polyfluoroalkyl substances (PFAS) represent a class of persistent organic pollutants found in furniture, cookware, home décor, clothing, firefighting foam, food packaging, and contaminants in food and water. PFAS are recognized as an environmental health priority by NIH due to their ubiquitous exposure, resistance to environmental degradation, and bioaccumulation. Growing evidence of their effects on vascular risk factors (e.g. hyperlipidemia, obesity, diabetes, hypertension) and neurotoxicity suggest that PFAS exposure increases Alzheimer’s Disease and Alzheimer’s Disease Related Dementias (AD/ADRD) risk, but empirical data are limited, weak, and inconsistent. Prospective cohort studies linking PFAS and AD/ADRD, with adjudicated clinical outcomes and control for confounding by diet and kidney function, are lacking. We propose to quantify the concentrations of 13 ubiquitous PFAS in archived serum samples from two time points, and the total PFAS exposure burden, in the Northern Manhattan Study (NOMAS, N=1290), an established (25+ year-long) multi- ethnic longitudinal cohort. We hypothesize that PFAS exposure increases the risk of cognitive impairment and AD/ADRD through a mechanistic pathway involving hyperlipidemia and atherosclerosis. PFAS are shown to alter lipid metabolism. NOMAS data have demonstrated a strong relationship between lipids and atherosclerosis. NOMAS participants had blood collected at baseline (1993-2001) and during follow-up (2003-2008), and annual follow-up, with comprehensive data on sociodemographics, vascular events, medical history, medications, health behaviors, diet, lipids, neural imaging, and a range of vascular risk factors. Participants had multiple comprehensive neuropsychological assessments, with extensive adjudication to identify those who developed AD/ADRD and mild cognitive impairment (MCI). NOMAS is ideal due to the diverse population (60% Hispanic, 20% Black, 20% White) at high risk for AD/ADRD and prospectively followed for vascular and cognitive outcomes. We will determine the associations between serum PFAS concentrations with comprehensive lipid profiles, carotid atherosclerosis phenotypes (plaque, intima-media thickness, stiffness), and risk of incident AD/ADRD and MCI, with the goal of identifying the impacts of PFAS on AD/ADRD, mediated through lipid metabolism and atherosclerosis. We will identify key confounders (diet), mediators (vascular risk factors), and effect modifiers (APOE4 genotype). The innovative strengths are the ability to fill important gaps related to the effects of PFAS on adjudicated AD/ADRD, ability to examine race/ethnic disparities in the effects of PFAS, and effect modification by APOE4, inclusion of a broad range of PFAS, and the interdisciplinary team with experience in PFAS exposure analysis and neuroepidemiology, which will lay the foundation for targeted community-wide preventive interventions. The data will provide crucial information on biological mechanisms through which PFAS impact cognitive impairment and AD/ADRD, which will inform public health recommendations on the avoidance of PFAS-containing products, and support regulatory efforts to reduce community PFAS exposure.

NIH Research Projects · FY 2025 · 2024-05

Project Summary Stroke is one of the leading causes of death and remains a major cause of disability with over 25 million individuals worldwide living with its consequences. Manifest atrial fibrillation (AF) accounts for 10% of all strokes, predominantly due to thromboembolism from the left atrium (LA). Despite thorough diagnostic evaluation, 20-30% of ischemic strokes are classified as cryptogenic or of unknown cause. Most cryptogenic strokes have an embolic appearance, known as embolic stroke of undetermined source (ESUS). A substantial portion of patients with ESUS will eventually be diagnosed with AF. Recent findings support the notion that an underlying disease of the LA (atrial cardiopathy), which may lead to AF, may also independently result in thrombus formation and increase stroke risk in the absence of AF. LA appendage (LAA) stasis is linked to atrial thrombus formation as part of Virchow’s triad. Our preliminary work on LA 4D flow MRI (4DMRI) provides a novel noninvasive tool that assesses LAA stasis and could provide a new paradigm for stroke prevention. Other biomarkers of atrial cardiopathy have already been associated with greater risk of development of AF and stroke, including atrial size/morphology, ECG P wave morphology, atrial strain on echocardiography, epicardial adipose tissue (EAT), troponin and N terminal pro-brain natriuretic peptide levels. Yet, none are currently used for clinical decision making. The overall goal of this research is to identify clinical and mechanistic characteristics of 4DMRI-derived LAA stasis in order to identify optimal strategies to prevent recurrent stroke in patients with ESUS. It is anticipated that this mechanistic assessment for risk of atrial thrombus formation will also provide improved utility for stroke prevention in AF. We will prospectively study patients with ESUS at 4 experienced centers with heart-brain MRI, assessing LAA stasis by 4DMRI and other atrial cardiopathy markers in a comprehensive manner. Specific aims are to : 1) Evaluate 4DMRI LA/LAA stasis and other atrial cardiopathy biomarkers in patients with ESUS (Group 1, n=229), cardioembolic stroke due to AF (Group 2, n=80), no stroke or AF (Group 3, n=80), and no stroke but with AF (Group 4, n=80). We will assess the diagnostic utility of LA/LAA stasis and other atrial cardiopathy biomarkers for cardioembolic stroke due to AF; 2) Assess the clinical factors predicting progression of LA/LAA stasis in 229 ESUS patients by repeating MRI at 2 years; 3) Determine whether LAA stasis and/or other atrial cardiopathy biomarkers are predictors of recurrent ischemic stroke (primary endpoint) and new AF (secondary) in 229 patients with ESUS. We hypothesize that LAA stasis will be the strongest independent predictor of recurrent ischemic stroke. This proposal, Cardiac ImagiNg of Embolic Mechanisms in Atriopathies causing Stroke (CINEMAS), focuses on optimizing the diagnosis of atrial cardiopathy in patients with ESUS. Improved diagnosis with a strong link to clinical endpoints can broaden the options for primary and secondary prevention of stroke and AF.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY Opioid Use Disorder (OUD) is a chronic, relapsing disease that, despite our best efforts to understand and mitigate the disease, has become a worsening public health crisis. 1,2 Currently, the medications to treat OUD interact directly with opioid receptors. 3 While this helps patients decrease the use of more dangerous opioids, the mechanism of action of the currently available drugs continues to perpetuate the substance abuse cycle. Exploring novel non-opioid targets is necessary to overcome this current standstill. One specific alternative population we are interested in investigating is calcitonin gene-related peptide (CGRP) expressing neurons within the parabrachial nucleus (CGRPPBN). These neurons have been well studied in itch and pain fields and are known to project to the central amygdala (CeA) 4,5, a brain region directly implicated in processing the negative emotional valence associated with alcohol and opioid withdrawal. The objective of this proposal is to assess the functional role of CGRPPBN in opioid reinforcement and to determine the gene expression and epigenetic changes associated with opioid taking and opioid withdrawal. Our preliminary results show that CGRPPBN activity decreases during opioid taking and increases during opioid withdrawal. We have also found that this population of neurons robustly expresses the µ-opioid receptor along with glutamatergic markers, positioning this population as a potential direct activator of the CeA during opioid withdrawal. To further probe these results, I will inhibit CGRPPBN neurons using molecular and pharmacological approaches and determine the behavioral consequences of these interventions. One of the pharmacological approaches we will be testing is an FDA-approved CGRP inhibitor currently used for migraine treatment. We will then establish the molecular profile of these neurons using nuclear RNAseq and Cleavage Under Tagmentation (CUT&Tag) at baseline, as well as during morphine taking and protracted abstinence. By integrating transcriptomic data with epigenetic information on post-translational histone modifications, I will be able to build a comprehensive map of the active and inactive chromatin regions with their corresponding differentially expressed genes, which can be used to detect epigenetic targets critical to the identity of CGRPPBN neurons. The results of this study will allow us to establish the feasibility of targeting these neurons to treat OUD via currently FDA-approved medications or through novel epigenetic targets identified in this study. These results will broaden our understanding of withdrawal and help expand our therapeutic arsenal for the long-term maintenance of opioid abstinence and prevention of relapse.

NIH Research Projects · FY 2026 · 2024-05

Dysregulated inflammation contributes to retinal degeneration. Although inflammatory cells may initially limit retinal damage by phagocytosing dying mutant photoreceptors and secreting reparative molecules, excessive inflammation causes recruitment of additional reactive microglia, phagocytosis of viable photoreceptors and toxic cytokine release, which accelerates retinal degeneration irrespective of the primary mutation. Therefore, a central challenge for developing anti-inflammatory therapies for retinal degenerations is to selectively suppress detrimental inflammation while maintaining beneficial inflammatory responses. We recently demonstrated that the cytokine IL-27 regulates beneficial and detrimental inflammatory signals in the retina and protects photoreceptors. IL-27 is a pleiotropic cytokine that regulates tissue reactions to infection, neuronal disease and tumors. We found that higher endogenous IL-27 levels were associated with photoreceptor protection induced by an innate immunity inhibitor in the rd10 mouse model of inherited retinal degeneration. In studies directly testing IL-27 function, we demonstrated that intravitreal injections of IL-27 into rd10 led to reduced pro-inflammatory cytokines, increased rod and cone photoreceptor survival and prolonged improvement in visual acuity. Additionally, IL-27 increased markers of protective microglia and reduced neurotoxic microglia. Because most research to date has focused on pro-inflammatory signals in the retina, a major unanswered question is how anti-inflammatory regulators such as IL-27 reduce neurotoxic inflammation during retinal degeneration. This proposed study will build upon our exciting data by defining mechanisms by which IL-27 signaling regulates inflammation and promotes photoreceptor survival (Aim 1). Additionally, IL-27 is expressed in Muller glia and IL-27 receptors are in microglia but whether these cells coordinate to regulate inflammation through IL-27 is not known. We will use conditional knockout mice and gene overexpression to identify whether a novel Muller glia and microglia cytokine-cytokine receptor signaling axis mediates IL-27-induced photoreceptor protection (Aim 2). The research team combines the expertise of the PI, Abigail Hackam – retinal degeneration and neuroinflammation, and collaborator Stefan Kurtenbach – scRNA seq and transcriptomics. The findings from this study will be significant to identifying a novel regulatory signal that controls inflammation, which will be relevant for precisely controlling inflammation and restoring tissue homeostasis during retinal degeneration. We will also generate novel insights into the roles of Muller glia-microglia interactions in controlling retinal inflammation. Finally, by characterizing a potential new therapeutic approach that is not dependent on the primary mutation, this study will set the foundation for developing IL-27 as a therapeutic strategy for protecting photoreceptors and rescuing vision during retinal disease.

NIH Research Projects · FY 2025 · 2024-04

Project Summary/Abstract Delivery of potent and broadly neutralizing antibodies with recombinant adeno-associated virus vectors (AAV) is a promising approach for both, the prevention and treatment of HIV. Recombinant AAV has an outstanding safety record in over 200 clinical trials and gene delivery mediated by AAV can result in continuous durable expression of the transgene product. However, because of prior exposure to naturally circulating wild-type AAVs, a significant portion of the human population has antibodies to the AAV capsid proteins in circulation. These preexisting capsid antibodies can prevent cell transduction after recombinant AAV vector administration, jeopardizing the applicability of this approach. To overcome this crucial issue, we propose a set of approaches aimed at exerting a profound but transient depletion in vivo of circulating IgGs in AAV-positive macaques (Aim 1). AAV-mediated delivery of an anti-HIV antibody will then be assessed during such transient depletion. Our goal is to obtain consistent and robust AAV-mediated delivery of anti-HIV antibodies with independence of the capsid serostatus. If successful, the proposed strategies to overcome preexisting AAV antibodies could be a key factor in the development of future AAV-gene delivery applications and more specifically in the fight against HIV. By overcoming preexisting AAV capsid antibodies, we aim at making the AAV-delivery of antibodies a safe and reliable approach against HIV. If satisfactory delivery methods are found, it becomes possible to envision a) long-term control of the viral loads in the absence of antiretroviral treatment by delivering a combination of potent and broadly neutralizing antibodies in people and b) long-lasting protection when this approach is used in a prophylactic setting.

NIH Research Projects · FY 2026 · 2024-04

Paclitaxel is a key drug used in several solid tumors including ovarian cancer, used alone or in combination with carboplatin. While often highly effective, paclitaxel has major adverse effects, namely sensory neuropathy and severe hair loss (alopecia). Since alopecia is psychologically devastating for many patients, preventing paclitaxel-induced alopecia is a major, as yet unmet need in clinical oncology with a massive impact on quality of life for cancer patients. Paclitaxel causes hair loss by stabilizing microtubules in the highly proliferative hair matrix keratinocytes, resulting in apoptosis, hair follicle dystrophy, and obliteration of the hair follicle’s capacity to regenerate. While hair loss may be reduced by scalp cooling to reduce drug exposure in some patients, the benefit is limited and unpredictable. Therefore, the development of satisfactory methods to reliably prevent paclitaxel-induced alopecia is an urgent, unmet medical need. We have made the unexpected discovery that a brief exposure to low intensity ultrasound can effectively neutralize the cytotoxic effects of paclitaxel on cultured cells by disrupting paclitaxel-induced rigid microtubule bundles, thus preventing cell death. Therefore, we propose here to rigorously test whether low intensity ultrasound can prevent the hair follicle cytotoxicity of paclitaxel in both ex vivo human scalp hair follicle organ culture and in vivo mouse models. Specifically, we shall study how ultrasound impacts on the microtubule network of hair matrix keratinocytes and hair follicle epithelial stem cells, and whether it can prevent paclitaxel cytotoxicity ex vivo and in vivo. Consequently, we plan to develop an approach using low intensity ultrasound to prevent paclitaxel-induced hair loss in mice. Preliminary results validated the experimental systems and support the hypothesis that a brief exposure of low intensity ultrasound is able to remove paclitaxel cytotoxicity in both human scalp hair follicles and mouse epidermal hair follicles. The ultimate goal is to translate the results into a clinical protocol designed to utilize ultrasound to prevent paclitaxel-induced hair loss in cancer therapy using paclitaxel and other microtubule stabilizing agents. Since low intensity ultrasound has been widely used in medicine with an excellent safety profile, the development of ultrasound treatment to prevent paclitaxel-induced alopecia is highly practical and feasible. Our study will provide proof-of-principle and critical guidance for the optimized utilization of ultrasound therapy to effectively prevent alopecia, thus greatly improving the quality of life of cancer patients.

NIH Research Projects · FY 2026 · 2024-04

Title: Specificity, Phenotype and Function of Pancreatic CD8 T Cells in Human Type 1 Diabetes Project Summary Human type 1 diabetes (T1D) is characterized by the immune-mediated destruction of insulin-producing pan- creatic beta cells. CD8 T cells are the most common cell found in insulitis lesions and are the principal T cell type implicated in beta cell destruction. Insulin and its precursors have been identified as key autoantigens in humans and mice due to their local abundance. The previous funding cycles for this grant have enabled us to lead the field of human islet cell investigation in T1D. Using samples from the national pancreatic organ donor consortium (nPOD) we have detected, precisely quantified, and identified the exact location of autoreactive pre-proinsulin (PPI) specific CD8 lymphocytes within the pancreata of human donors. Studies performed within this project have already revealed significant and novel findings. We first identified auto-reactive CD8 T cells in the islets of patients with T1D using a specially designed method of tetramer staining. We found that their frequency is high and likely the most predominant autoreactive cell type (1), a finding which has been inde- pendently confirmed with other methods by Nakayama et al. (2). More recently (3) we defined the precise location of these cells within a pseudo-timeline of disease development and concluded that PPI cytotoxic T lymphocytes (CTL) are already present in healthy pancreata and appear to become 'attracted' to islets during the development of T1D. This seminal observation indicates that the islets appear to expose themselves to immune recognition, and thus become a key culprit in driving T1D development. A new key development of this grant is the Orion (Rarecyte) technology that allows for highly multiplexed studies in whole tissue sec- tions, combined with state-of-the-art analysis software. The study of the whole section and multiple targets simultaneously allows to define the islet/immune interface and the communication between PPI CTL and other immune cells more precisely and with a much reduced number of samples. The present proposal will define the precise location and function of these interactions, and corroborate these findings in highly innova- tive human islet microtissues and living donor pancreatic tissue slices using dynamic in vitro models. Finally, this proposal will decipher mechanistically how some aspects of the islet immune interface can be reset/restored by the action of GLP1 agonists, building on our recent work which has shown that this drug class can maintain beta cell function (glucose-induced insulin secretion) in vitro and in vivo during the devel- opment of T1D (4). Overall, our findings should give us an unprecedented and unique understanding of how and why T1D develops, and provide mechanistic information using our novel in vitro systems, thus ultimately aiding the development of new therapeutic options.

NIH Research Projects · FY 2025 · 2024-04

ABSTRACT In the United States, it is estimated that more than 9% of people living with HIV acquired the virus through injection of substances or substance abuse-related sexual contact. Although substance use is well known to exacerbate HIV-associated neurotoxicity, there is no prompt treatment for alleviating neurocognitive dysfunctions in HIV-positive individuals. Our proposal is based on significant discoveries regarding the impact of chronic METH and HIV on the neurogenic niche, such as (i) HIV can infect the neural progenitor cells (NPCs) in subventricular zone (SVZ) of the lateral ventricle in mouse brain; (ii) chronic METH and HIV changes microRNA (miRNA) profiles of ex vivo NPCs; and (iii) METH and HIV-disrupted blood-brain barrier (BBB) integrity increases brain inflammation, but it does not stimulate the migration of NPCs in that lesion. The overall goal of this proposal is to investigate a novel mechanism linking the BBB pericytes and NPCs in chronic METH and HIV-induced alterations of NPC migration mediated by miRNAs. In order to evaluate this hypothesis, our transdisciplinary study will focus on the role of miR-21-5p, whose expression is significantly reduced in METH and HIV-exposed mouse SVZ NPCs and plasma samples, in the alteration of SVZ NPCs migration (Aim 1), and the crosstalk between the BBB pericytes and NPCs in response to METH exposure and HIV infection in the brain (Aim 2). We anticipate that this proposed research will demonstrate the potential of restoring the NPC migration to combat the neurotoxic effects of METH and HIV, ultimately improving neurocognitive function by enhancing adult neurogenesis. The focus on the interaction between the brain microvasculature and NPC migration, which influences the development of neurocognitive dysfunctions following chronic METH exposure and HIV infection, is an innovative and cutting-edge conceptual approach. Moreover, given the growing evidence linking miRNA expression alterations to substance abuse and HIV infection, studying the role of miRNAs in METH and HIV-induced altered neurogenesis is crucial and provide new insights into the underlying mechanisms of neurotoxicity and potential therapeutic targets.

NIH Research Projects · FY 2025 · 2024-04

SUMMARY Cells use a broad arsenal of proteins to change the curvature of cell membranes. This is important for cell motility, endocytosis, and establishment of cell morphology. As such, it comes as no surprise that components of this machinery are central in the etiology of many diseases, including developmental and immunodeficiency disorders, neurodegenerative conditions, and cancer. One protein family that appears repeatedly at the intersection of membrane biology and disease is the Bin1/Amphiphysin/Rvs family of proteins (BAR proteins). Homologues of this family are found from yeast to humans and are essential for the induction of morphological changes of cells and for the coupling of membrane curvature to actin polymerization. While BAR proteins are proposed to function in these processes, little is known about the structure/function of these proteins in an in vivo context. This gap of knowledge hinders leveraging BAR proteins and their targets for the treatment of cancer and other diseases. A comprehensive model of full-length BAR proteins is needed to perform structure/function analysis in vivo, so physiologically relevant structures can be identified. The goal of this exploratory proposal is twofold: to establish an in vivo model of BAR protein structure/function using the pioneering organism C. elegans (Aim 1), and to use cryoEM/cryoET to identify salient structural motifs in a full-length human BAR protein bound to the membrane in the presence and absence of one of its main targets, the small Rho GTPase CDC42 (Aim 2). The accomplishment of these two aims will provide information on essential structural interfaces and amino acid residues that can be then tested in vivo. Data obtained here will set the groundwork for a comprehensive in vivo determination of BAR protein structure/function that we will pursue in the future. Ultimately, this work will advance our understanding of how cells change morphology in health and disease.

NIH Research Projects · FY 2025 · 2024-04

Project Summary Alzheimer's disease (AD) is a major public health concern in the US, and there is an urgent need for reliable, inexpensive, and non-invasive biomarkers to identify individuals at risk for AD. This project aims to develop innovative computational strategies for precision medicine in AD. Specifically, we will harmonize several large longitudinal clinical datasets, identify DNA methylation (DNAm) biomarkers for cognitive reserve (CR), and build DNAm-based prediction models for AD. In this project, we will improve the accuracy of DNAm-based prediction models by leveraging knowledge of cognitive reserve, harmonizing multiple datasets, and training and testing prediction models using samples from longitudinal studies. The DNAm-based prediction models will provide an inexpensive and convenient approach for identifying subjects most likely to progress to clinical AD, reducing heterogeneity in patients selected for clinical trials, and facilitating personalized treatment strategies in AD. Additionally, identifying DNAm markers for CR will help develop novel therapeutic targets and lifestyle interventions for preventing dementia. The successful completion of the project will also provide the community with harmonized datasets for AD research, as well as computational methods and tools that can be easily adapted and applied to analyze datasets in other types of dementia.

NIH Research Projects · FY 2026 · 2024-03

ABSTRACT Depression is the most prevalent neuropsychiatric condition in IV infection and frequently associated with substance use disorders (SUD), including methamphetamine (METH) use disorder. Nevertheless, there is an alarming lack of information on the interactions between HIV, SUD, and depression, emphasizing a critical gap of knowledge on the mechanisms of HIV and/or METH impact on the development of depression. The present application is focused on the role of the inflammasome in this disease. Based on our preliminary results, we formulated the central hypothesis that HIV and/or METH-induced inflammasome activation primes for the development of depression. Mechanistically, we will focus on the dysregulation of the gut-brain axis in this process because the gut microflora is a potent activator of the inflammasome and a producer of a variety of mood-influencing metabolites and neurotransmitters. Moreover, there is strong recent evidence on the involvement of chronic neuroinflammatory responses and the disruption of the blood-brain barrier (BBB) influencing the development of depression. We propose to evaluate the following sequence of events in a novel humanized mouse model infected with HIV and exposed to METH: HIV infection/METH exposure → inflammasome activation → dysfunction of tissue barriers (the gut and the BBB) → neuroinflammation → depression. Successful completion of this proposal will meet the translation objective, which is to demonstrate that HIV infection and/or METH exposure contributes to, and accelerates, the pathomechanisms of depression via the alterations of the gut-brain axis, which involves inflammasome activation and chronic brain neuroinflammatory responses. The significance of our proposal is within its focus on the leading public health problem in people living with HIV (PLWH), namely depression. The mechanisms of HIV and/or METH-induced development of depression are largely unknown, making the proposed studies innovative and likely to generate unique data sets. The resulting discoveries may have significant epidemiological, economic, and social implications. Knowledge of the underlying mechanism(s) whereby HIV and METH prime individuals to depression may provide critically important and therapeutically relevant information to identify novel targets for pharmacological intervention. Our long-term goal is to characterize how HIV and METH contribute to the pathomechanisms of depression and, ultimately, prevent its development through a precision therapeutic intervention.

NIH Research Projects · FY 2026 · 2024-03

Project Summary/Abstract Background: This fellowship application is from a second-year doctoral student seeking to establish expertise in HIV among immigrant Latino men who have sex with men (LMSM), adaptation science, and implementation science (IS). Miami-Dade County (MDC), Florida, is the project's research site, a priority jurisdiction under the Ending HIV Epidemic (EHE) plan, where the HIV burden is among the highest in the United States, particularly among LMSM.1 In MDC, unemployment, financial stress, and poverty contribute to HIV vulnerability among LMSM, especially recently arrived immigrant LMSM.2–4 Interventions are needed to address these structural barriers that impede the success of evidence-based biomedical HIV prevention and treatment tools such as pre-exposure prophylaxis (PrEP) and treatment among immigrant LMSM. Work2Prevent (W2P) is one evidence-based structural intervention which has shown to impact HIV prevention outcomes among English- speaking Black and Latino MSM and transgender women, respectively.5–8 However, W2P has yet to be adapted for immigrant LMSM populations to address their unique needs, nor for people living with HIV. Research Strategy: This fellowship would be the first to adapt an HIV-status neutral structural employment as prevention intervention (W2P) to address the structural barriers for LMSM with recent immigration histories who may be monolingual Spanish-speaking, or undocumented. This will be accomplished by obtaining feedback from immigrant LMSM (n=10) and topical experts (n=10). The research project, linked to a carefully considered training plan, will advance the candidate's trajectory of becoming an independent investigator in the science of adaptation, immigrant LMSM HIV disparities, and implementation science. The proposed project aligns with the co-primary sponsor's NIH-funded research (PI: Harkness K23MD015690, P30MH116867-03S1) to scale up and disseminate PrEP, HIV testing and behavioral health services to LMSM in South Florida. The following aims are proposed: Aim 1 will qualitatively determine key adaptations to make W2P culturally grounded for recent immigrant LMSM and status neutral via theater tests. Aim 2 will elucidate further adaptations to W2P and plan for implementation based on topical expert input through focus groups. ADAPT-ITT,9 an established framework for adapting interventions, will guide the systematic adaptation process. Training Plan: The training plan, composed of coursework, seminars, and individual meetings, will allow for developing the necessary skills to achieve the research aims and expertise in culturally adapting and enhancing the feasibility of implementing evidence-based interventions that address structural factors driving HIV disparities among immigrant LMSM. Next Steps: Study findings will inform a subsequent K01 proposal to pilot test the culturally adapted W2P program for immigrant LMSM. The subsequent K01 will fully launch PI Jaramillo to research independence. Innovative methods in adaptation and IS and focus on the unique needs of immigrant LMSM are urgently needed in geographic hotspots to achieve health equity and EHE goals.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY/ABSTRACT In response to RFA-DA-21-250, we propose to investigate inflammation, blood-brain-barrier (BBB) permeability and reward functions in people with HIV (PWH) and comorbid depression. Depression is the most common neuropsychiatric illness among PWH, with an average prevalence of up to 78% in some cohorts. Alarmingly, it is estimated that by 2030, the top two leading causes of disease burden globally will be HIV and depressive disorders. These data highlight the urgent need for research focusing on neurobiological mechanisms underlying HIV/depression comorbidity. Our proposal addresses this need. Our proposed model: (1) HIV infection induces systemic inflammation [peripheral blood mononuclear cells (PBMC), cytokines]; (2) systemic inflammation extends to the CNS through transmigration of PBMC subtypes through the BBB; (3) disruption of BBB integrity and neuroinflammation lead to alterations in the reward circuitry, contributing to depression in PWH. In support of this model, our group has pioneered the study of BBB in PWH, establishing a highly reproducible and reliable in vitro model of the human BBB, comprised of a co-culture of human brain microvascular endothelial cells and human astrocytes. We showed that compared to healthy controls (HC), specific PBMC subtypes from PWH preferentially transmigrate across the BBB model, despite suppressed viral load. In our depression research, we found that anhedonia–a core symptom of depression reflecting reward deficits–was associated with worse depression outcomes, including chronicity and suicidality. To better delineate reward circuitry, we identified distinct resting-state network features associated with depression and anhedonia using striatal-based intrinsic functional connectivity and whole-brain parcellation data-driven graph theory analysis. We additionally utilized the reward flanker (RFT) and reward prediction error (RPET) fMRI tasks to examine distinct brain activity during reward anticipation, attainment, and prediction errors. Furthermore, we reported associations between circulatory cytokines with both anhedonia and reward neurocircuitry in youth. In addition, our team has implemented dynamic contrast-enhanced (DCE) MRI and a water-extraction-with-phase-contrast-arterial-spin- tagging (WEPCAST) MRI, enabling in vivo regional and global BBB permeability, respectively. Extending our compelling findings expertise, we will test the overall hypothesis that PWH exhibit increased systemic inflammation and BBB disruption (assessed in vivo and in vitro), leading to reward dysfunction and depression. We will utilize a 2×2 factorial design: 1) 100 depressed PWH; 2) 100 non-depressed PWH; 3) 50 depressed HIV negative people; and 4) 50 HC. We will include subthreshold depression to capture a wide range of depression severity. Study procedures will assess psychopathology, reward, anxiety, trauma, cognition, HIV treatment, CD4+ count, viral load (VL), and immune assays. Neuroimaging will include DCE-MRI, WEPCAST and fMRI.