University Of Miami School Of Medicine

NIH Research Projects · FY 2025 · 2025-09

Background: Jump Start to Talk aims to understand culturally embedded perspectives of caregivers and early care and education (ECE) teachers from Spanish- and Haitian Creole-speaking communities regarding late talkers (LTs) who are dual language learners (DLLs). Using a dual-community comparative approach to examine both Hispanic and Haitian populations, this study addresses critical gaps in understanding LTs who face risks of persistent language delays and poor academic outcomes, heightened for DLLs due to misidentification and cultural-linguistic service access barriers. Approach: Using Israel et al.'s community-based participatory research (CBPR) approach, we will leverage partnerships between the University of Miami and three local organizations: Sant La Haitian Neighborhood Center (urban), Le Jardin Hispanic Community Center (rural), and Speech Pathology Educational Center (SPEC), serving families in both communities. We will co-design and co- implement the study with these community partners and their cultural brokers - trusted community members who share participants' linguistic and cultural backgrounds. Through purposive sampling of our network of 75 ECE centers serving Hispanic and Haitian families, we will identify 30 LT DLLs using culturally and linguistically appropriate screening approaches. We will interview their caregivers and 30 ECE teachers. Applying Sapiets et al.'s service access model and using the Framework Method for qualitative analysis, we will co-analyze interview data with our community partners to understand factors influencing service access from identification to provision, addressing two aims: Aim 1: Recognition and Identification of Late Talking: What are the perceptions of caregivers and teachers from two underserved communities regarding a) how they recognize and understand typical versus delayed language development in DLLs, and b) what factors influence their decisions about seeking services? Findings will directly inform development of culturally and linguistically appropriate assessment tools for identifying LTs in DLLs. Aim 2: Early Support Service Access: What barriers and facilitators do caregivers and teachers from two communities experience when attempting to access and engage with early support services (ranging from preventive monitoring and classroom-based supports to formal early intervention programs, community-based services, and parent-implemented language support strategies) for DLL LTs? Impact: These aims meet a high-priority research area for NICHD PAR-24-046 by examining identification and referral processes for LT DLLs from Spanish- and Haitian Creole-speaking backgrounds. The innovative dual- community approach and use of cultural brokers will inform the development of culturally responsive screening and referral processes, particularly addressing the needs of Haitian Creole-speaking communities where validated measures are currently lacking. .

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Glioblastoma (GBM) is the most lethal primary brain tumor with a five-year survival rate of less than 5%. The GBM tumor microenvironment (TME) is highly immunosuppressive, conferring resistance to immunotherapies that have proven groundbreaking in treating other malignancies. Thus, the need to identify and target drivers of immunosuppression in the GBM TME remains critically important. Notably, our lab recently uncovered robust overexpression of Human Endogenous Retrovirus K (HERV-K) in GBM that is associated with poor patient outcomes. HERVs are remnants of ancient germ-line retroviral infections that comprise ~8% of the human genome. They are generally transcriptionally silenced, but dysregulated states like cancer can lead to increased expression of canonical HERV genes and proteins, including env (envelope protein). This project proposes HERV-K Env as a viable candidate to target therapeutically to improve GBM patients’ response to immunotherapy due to robust evidence that retroviral Env proteins have immunosuppressive properties and contain a highly conserved immunosuppressive domain (ISD). Recent results, including our preliminary data, show that HERV-K Env inhibits proliferation of T cells, modulates cytokine secretion, and increases expression of immune checkpoint receptors. Further, immune deconvolution of bulk RNA sequencing and spatial transcriptomics show that high HERV-K Env expression is associated with low immune cell infiltration in GBM and decreased survival of GBM patients. However, the mechanism by which HERV-K Env suppresses T cell function remains elusive, and HERV-K Env’s exact role in the immunosuppressive GBM TME remains unclear. This proposal will test the central hypothesis that HERV-K Env’s ISD mediates binding to critical surface receptors, including CD98hc (roles in amino acid transport and integrin signaling) on T cells, altering their downstream functions and leading to T cell dysfunction and to greater immunosuppression in the GBM TME. Specific Aim 1 will identify critical mediators of HERV-K Env’s immunosuppressive effect on T cells via the use of co-immunoprecipitation and mass spectrometry to identify the binding partners of Herv-K Env on the T cell surface and to elucidate the ISD’s role; further, it will explore alterations in downstream pathways of these receptors via methods including Western blot, qPCR, flow cytometry, and ELISA. Specific Aim 2 will characterize HERV-K Env-mediated immunosuppression in the GBM TME by utilizing GBM-T cell co- cultures, HERV-K Env-overexpressing orthotopic mouse models paired with single cell RNA sequencing and multiplex flow cytometry, and spatial transcriptomics analysis of patient specimens. Altogether, this study’s results will explain critical immunosuppressive mechanisms of a retroviral protein, HERV-K Env, in GBM and elucidate its role in GBM’s highly immunosuppressive TME. It possesses robust translational potential by providing a candidate to target therapeutically and by identifying key pathways in Env’s immunosuppressive effect in GBM.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Chronic itch is a considerable medical challenge associated with various skin and systemic diseases, , exerting a profound impact on patients' overall quality of life. While the epidermis is conventionally considered the source of skin itch, critical knowledge gaps persist regarding the role of Langerhans cells (LCs), resident cells in the epidermis, in mediating itch. LCs play a vital immunologic role in protecting against environmental antigens and, due to their dendrite-like appearance and proximity to nerve endings, have been associated with the nervous system. This project aims to comprehensively understand the contribution of LCs to chronic itch, shedding light on a perplexing condition. LCs can be categorized into resident and monocyte-derived types. We hypothesize that resident LCs are implicated not only in acute itch but also in chronic itch. Aim 1 will examine the interaction of LCs with pruriceptors in mouse models of chronic itch. Utilizing transgenic mice expressing TdTomato in pruriceptors, we will visualize the localization of pruriceptors and LCs in chronic itch mouse models such as atopic dermatitis, psoriasis, and postburn. Optogenetic stimulation of LCs will be employed to determine if it activates pruriceptors. Aim 2 will evaluate the contribution of resident and/or monocyte-derived LCs to itch in mouse models of chronic itch. Utilizing hematopoietic chimeras with Gq- DREADD and Gi-DREADD expression, we will investigate the roles of resident and monocyte-derived LCs in spontaneous itch in chronic conditions. Chemogenetic activation and inhibition of these LC subsets will be explored to understand their influence on chronic itch. Aim 3 will identify and genetically define the heterogeneous population of resident and monocyte-derived LCs in mouse models of chronic itch. Single-cell RNA sequencing and cell sorting techniques will be employed to define the heterogeneous population of both LC types and examine how their dynamics change in mouse models of chronic itch. This comprehensive investigation seeks to unveil the overlooked role of LC-pruriceptor signaling in promoting spontaneous itch under diverse chronic conditions. The long-term goal of this study is to identify functional LC subsets, comprehend their impact on chronic itch, and pave the way for the development of more effective, mechanism- based treatments for this significant healthcare challenge.

NIH Research Projects · FY 2025 · 2025-08

Gliomas arise from neural progenitor cells (NPCs) or their descendants that derive transcriptional identity from aberrant chromatin landscapes. Polycomb repressive complex 2 (PRC2) is a crucial chromatin modifier that orchestrates gene silencing and organizes genome architecture during lineage specification and commitment by depositing repressive histone H3K27 trimethylation marks. While mechanisms that alter PRC2 chromatin occu- pancy are understudied, they hold significant conceptual appeal due to the determinative role of PRC2 in gov- erning the fate of NPCs. We have shown that IκBα deletions frequently occur in diffuse gliomas, portend poor prognosis, reshape the DNA and histone methylome antipodal to the IDH mutation, and induce a transcriptome landscape partly resembling that caused by PRC2 loss-of-function in H3K27M mutant pediatric gliomas. IκBα is the main regulator of nuclear factor-κB (NF-κB), but we found phosphorylated-SUMOylated (ps-)IκBα exerts an alternative chromatin-regulatory function by dynamically binding histones H2A and H4, thereby recruiting PRC2 and priming lineage choice. The scientific premise of this proposal is grounded in our discovery that IκBα exerts a novel PRC2-regulatory role in glioma and progenitor cells, which is independent of NF-κB. Defining the PRC2 vs. NF-κB-dependent roles of IκBα is pivotal for understanding their malfunction during lineage specification and gliomagenesis. Our central hypothesis is that IκBα recruits PRC2 to active chromatin and thus controls critical lineage programs that are aberrant in gliomas. Our specific aims are: 1. Define the Contribution of Chromatin vs NF-κB Specific IκBα Functions in Neural Lineage Specification. We will use human induced pluripotent stem cell (iPSC)-derived IκBα-deficient (CRISPR/Cas9 RNP DIκBα)/Tet-On:IκBα-SOF (SOFDNF-κB vs. SOFDH2A/H4) tripotent wildtype iNPCs and their lineage-committed progeny to determine whether, independent of NF-κB, chromatin-bound ps-IκBα regulates chromatin architecture and modulates the transcription of developmental genes during neural lineage specification and commitment. 2. Define the Mechanism of Dynamic PRC2 Reg- ulation by IκBα. We will utilize wildtype iNPCs with DIκBα/Tet-On:SOFDNF-κB vs. DIκBα/Tet-On:SOFDH2A/H4 geno- types to define the dynamics of IκBα binding to PRC2-regulated chromatin regions and determine its impact on recruitment and assembly of PRC2 subcomplexes (PRC2.1/PRC2.2), characterized by distinct accessory pro- teins, which differ in their extent of poised lineage-specific gene silencing. 3. Define the Consequences of IκBα Editing on Fate-Switches and Gliomagenic Trajectories from NPC to GBM. We will use a gliomagenic (DCDKN2A/B-PTEN-TERTmut) derivative of the iNPC model genome-edited to carry mutations governing vari- ous iGBM subtypes (EGFRvIII/PDGFRAdel8/9/DNF1) and GBM PDXs, to define the impact of DIκBα/Tet-On: SOFDNF-κB vs. DIκBα/Tet-On:SOFDH2A/H4 on chromatin and PRC2 regulation, multi-step reprogramming, and fate- transitions driving gliomagenic trajectories from NPCs to GBM. Our studies will comprehensively elucidate the molecular dynamics of lineage-specific chromatin regulation by IκBα and its subversion during gliomagenesis.

NIH Research Projects · FY 2025 · 2025-08

Modified Project Summary/Abstract Section This Type 3 Hybrid Effectiveness-Implementation study will evaluate PrEP & Soccer, a multicomponent and multilevel implementation strategy delivered by a pharmacy chain to increase PrEP knowledge and initial PrEP appointment attendance among PrEP-eligible men. Study Context: The study will be conducted in Miami-Dade, FL, a priority jurisdiction with high HIV rates, particularly among men. Pharmacy chains provide a neutral setting conducive to expanding PrEP knowledge and access. Implementation Strategy: Monitoring & Evaluation: Quality monitoring systems will ensure effective study progress. Colleague Engagement: CVS executives and store managers will review study progress and collaborate on adjustments. Pharmacy Staff Sensitization: Educational materials will train pharmacy staff on HIV prevention and the importance of PrEP for the Miami-Dade community. Sustaining the Strategy: An implementation playbook will be developed to guide the scale-up and sustainability at CVS Health and other pharmacy chains. Streamline Recruitment: PrEP navigators will streamline client enrollment in pharmacy settings. SAFE Training: Our community partner will train PrEP navigators on HIV prevention and client-tailored PrEP navigation. Personalized PrEP Navigation: Client-tailored navigation will promote PrEP knowledge and initial appointment. Insurance & Patient Assistance: Navigators will assist with PrEP cost coverage for all participants (insured, underinsured and uninsured). Marketing Plan: Marketing flyers and an audio series will engage men in PrEP & Soccer. Study Design: Using a parallel cluster randomized controlled design, eight CVS Health pharmacy locations will be assigned to PrEP & Soccer or a general CVS Health service information-only control (four stores in each arm). A total of 800 participants will complete quantitative assessments at baseline, 3- and 6- month follow-ups, while 144 participants will also engage in qualitative interviews. Implementation Aims: Aim 1a. Evaluate PrEP & Soccer’s Reach (extent to which persons at elevated HIV vulnerability initiate participation), and Implementation (extent to which implementation strategies are delivered as designed). Aim 1b. Examine contextual factors associated with Reach and Implementation (e.g., facilitators and barriers; participants’ reactions; what works for whom under what circumstances). Effectiveness Aim: Aim 2. Evaluate effectiveness of PrEP & Soccer on initial PrEP appointment attendance (primary effectiveness outcome), positive movement along the PrEP cascade, and PrEP knowledge (secondary effectiveness outcomes) compared to control. Sustainment & Scale-up Aim: Aim 3. Assess the maintenance, sustainment, and scale-up potential for the strategies by determining (1) cost and needed resources, (2) intent and strategies to sustain and scale using Evidence Based Quality Improvement (EQBI) process, and (3) development of an implementation playbook.

NIH Research Projects · FY 2025 · 2025-08

Recent studies have demonstrated the critical importance of including individuals of varied backgrounds in complex trait research to ensure broad research benefits and comprehensive understanding of diseases like Alzheimer Disease (AD) and AD-related dementias. The GAP-ADN Consortium aims to address this need through several key initiatives: coordinating consortium activities, promoting collaboration across the AD/ADRD research community, facilitating training opportunities, and coordinating data sharing. Through these efforts, the consortium will provide critical training opportunities for early-stage investigators while enabling senior investigators to expand their research globally, ultimately working to ensure that research progress in low- and middle-income countries (LMICs) is fully integrated with ongoing AD/ADRD research initiatives. The goal of this proposal is to establish a Coordinating Center to support and enhance collaboration among AD/ADRD researchers in low- and middle-income countries (LMICs) through administrative coordination, partnership building, training opportunities, and data sharing mechanisms. The necessity of including individuals of varied ancestry in studies of complex traits has been clearly demonstrated, particularly in Alzheimer's disease where risk factors like the ApoE4 allele show differential effects based on ancestral background. Many US populations of various backgrounds trace their lineage to LMICs, making it critical that researchers in these countries have the tools and infrastructure to conduct high-quality research. However, LMIC investigators face significant obstacles in carrying out their studies. Drawing on our extensive experience coordinating large international consortia, several with LIMC collaborators, we will establish a Coordinating Center, Global Alzheimer’s and Dementia Partnership to Advance Neuroscience in LMICs (GAP- ADN), that will: 1) Provide comprehensive administrative and regulatory oversight while facilitating consortium- wide activities and reporting, 2) Promote collaboration and communication through website development and partnership building across the global AD/ADRD research community, 3) Create training opportunities for both early-stage and senior investigators to promote global participation and independence in AD/ADRD research, and 4) Coordinate data sharing and resource dissemination through NIA-approved repositories and consortium resource hubs following FAIR principles. Our team brings deep expertise in global research administration, AD/ADRD science, infrastructure building, and consortia management. The Coordinating Center will leverage this experience to break down obstacles between established and emerging investigators while ensuring that research benefits and infrastructure development reach all populations affected by AD/ADRD. By enabling broad participation in AD research while creating sustainable research networks between investigators globally, this initiative will help build crucial capacity for understanding the complete architecture of AD/ADRD across all populations.

NIH Research Projects · FY 2025 · 2025-08

Pneumonia (PNA) is one of the leading causes of death worldwide. PNA can result in devastating acute inflammatory injury in the lung manifesting in acute respiratory distress syndrome (ARDS). Current treatments for PNA have focused on the pathogens, but do not target excessive lung inflammation elicited by the host immune response. Both the emergence of new infections, typified by COVID-19, and the expanding impact of antimicrobial resistant pathogens, highlight the limitations of our current armamentarium and underscore the need to identify additional therapeutic targets in PNA-induced ARDS. With the understanding that resolution of PNA is an actively regulated program to promote return to homeostasis, our work has focused on identifying cellular and molecular mediators of this resolution phase. Others and we have demonstrated that regulatory T cells (Tregs) promote resolution of infectious-ARDS. Our strong preliminary data has identified lung-derived Treg DHX58, which encodes an RNA helicase protein essential for antiviral responses, as a candidate gene upregulated during the resolution phase of ARDS. DHX58- deficient animals fail to resolve lung inflammation after Streptococcus pneumoniae-ARDS with significantly diminished lung Treg numbers during injury resolution, implicating DHX58 in optimal Treg function in vivo. Further, we observed significantly increased 30-day mortality among carriers of a putative loss-of-function variant of DHX58 with infectious ARDS (71% vs. 47%, p=0.01), underscoring the potential clinical impact of DHX58 in ARDS outcomes. Our in-silico analysis of the DHX58 promoter identified numerous estrogen responsive elements (ERE). Indeed, DHX58 expression was induced in Tregs by estradiol (E2). Importantly, our published work showed that therapeutic E2 promotes resolution of preclinical PNA-ARDS in a Treg-dependent manner. Estrogen receptor beta (ER) was necessary for both Treg-dependent rescue of lymphopenic hosts and Treg-mediated suppression of pro-inflammatory cytokine production in macrophages in vitro. Preliminary gene expression analysis and high- dimensional flow cytometry implicate E2 and its downstream-target, DHX58, in the regulation of critical Treg transcription factors (TFs), notably Foxp3 and GATA3. Thus, we hypothesize that E2, in part via ER-dependent upregulation of DHX58, orchestrates critical Treg pro-resolution functions, through regulating expression of key TFs in Tregs. The goals of this proposal are to determine the cellular, molecular and transcriptional determinants of E2-ER-DHX58 in Treg-mediated resolution of PNA-ARDS to provide the mechanistic underpinnings of the regulation and functional role of ER in Tregs.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract. Diffuse large B-cell lymphoma (DLBCL) is the most frequently diagnosed lym- phoma and carries poor prognosis for the ~40% of patients not cured by frontline therapy. While emerging im- munotherapies provide new options for relapsed or refractory (rel/ref) patients, kinase inhibitors have found little role in DLBCL management despite transformational impact on other lymphomas. In this proposal, we de- fine the cyclin-G associated kinase (GAK) as a novel therapeutic target, discovered through phenotypic screening and machine-learning target deconvolution. We find GAK is a cell-cycle kinase critical for mitotic spindles. DLBCL tumors with reduced function of the tumor suppressor retinoblastoma (RB) in particular show critical dependence on GAK. Preliminary data also demonstrate in vivo efficacy of GAK inhibition against DLBCL xenografts, employing a pharmacologically unfavorable tool compound. To leverage this discovery to rapid evaluation in rel/ref patients, we found clinically available kinase inhibitors with strong anti-GAK activities suitable for repurposing. Several of these have stronger activity against GAK than their primary clinical targets and demonstrate potent killing of DLBCL tumors. We also reveal phosphoproteomics that implicate for the first time direct substrates by which GAK regulates mitosis. Our central hypothesis is that rapidly dividing DLBCL cells require GAK’s phosphorylation of specific mitotic regulators for successful nucleation and positioning of mitotic spindles. To test this, we pursue three specific aims. Aim 1 assesses microscopically the context and mechanisms GAK kinase regulation of mitosis. Confocal microscopy optimized on a high-throughput live-imag- ing platform seek to define specifically the role of GAK kinase activity in mitosis and the mechanistic basis for increased GAK dependence in RB-deficient tumors. Aim 2 defines biochemically and functionally GAK’s phos- phorylation substrates that regulate mitosis. Substrates to be evaluated biochemically and functionally include MTUS2, CENPF, EZRIN, and other candidates from our preliminary data. Aim 3 evaluates anti-lymphoma effi- cacy and hematologic toxicity of GAK inhibition in vivo. Inhibitors of other mitotic kinases like Aurora kinase A have been plagued clinically by hematologic toxicities. Here, using complementary murine lymphoma models, we pursue a hypothesis that GAK inhibition will be efficacious at reduced toxicity. Moreover, results are de- signed to directly inform development of investigator initiated trials (IITs) of repurposed compounds from this project co-led by a lymphoma physician-scientist. We expect to define GAK – never before studied as a can- cer therapeutic target – as a critical dependency for mitosis by DLBCL. We both investigate mechanisms and rigorously determine GAK’s suitability as a novel clinical target. Our project is innovative because we show for the first time GAK is a mitotic kinase and the experimentally generated phospho-substrates by which it carries out these activities. Our project is significant because unmet clinical need remains persistently high for rel/ref DLBCL patients, and RB deficiency is a common, high-risk, and currently untargetable biomarker in patients.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract The purpose of this shared S10 instrumentation grant application is the purchase of a Leica upright Stellaris 5 confocal microscope to replace an obsolete, end-of-life Leica SP-5 confocal microscope dedicated for live animal imaging. The main aim is to continue to provide high-resolution intravital confocal imaging capabilities to the University of Miami UM research community by upgrading and replacing the existing instrument (now 18 years old) that has reached the end of its life and can no longer be repaired or serviced. The requested microscope fills a critical need and will support the ongoing efforts of the NIH-funded research of 9 Major User groups representing 5 Departments at the University of Miami (UM). Other than the microscope we are replacing, there are no other available, open access microscopes at UM configured for live animal imaging studies that meet the needs of the existing and future User Groups. The replacement Leica Stellaris 5 system is configured with a white light laser (WLL) having an excitation range from 495 to 790 and a detection range up to 850 nm. It also has a 405 nm laser for near-UV excitation. The extended excitation wavelengths provided by the WLL allows for additional fluorescent reporters to be imaged simultaneously in a single experiment, thereby providing additional information compared with the older technology that was limited to the detection of 4 or 5 fluorescent reporters at best. The Stellaris 5 system enabling the multiplexing of fluorescent reporters in a single experiment also reduces the utilization of research animals consistently with the 3Rs of animal research (Replacement, Reduction, Refinement). The microscope is equipped with a 5x and a long- distance 10X dry objects and with apochromatic water-immersion objectives (16x, 25x, and 40x) with high numerical aperture and large free working distances that are essential for intravital imaging. The upright configuration of the objective lenses is required for proper access to the live animal tissue to be imaged and positioning of the objectives to regions of interest. The upright configuration and long working distance objectives are necessary to accommodate the anesthesia apparatus and stereotaxic head-holder, which require marked clearance from the microscope body and objectives mounting turret to allow positioning of the animal on the stage and under the objective. The instrument will be housed and maintained at the open access Analytical Imaging Core Facility (AICF), where the entire UM community has access to instrumentation on a fee-for-service basis. The existing confocal microscopes in the AICF and other imaging facilities at the University, other than the obsolete Leica SP-5 system, allow for high precision state-of-art imaging of ex vivo tissues and live cells in culture vessels; however, the inverted configuration of those systems and the objective lenses they are equipped with are not amenable for live animal imaging, for the reasons elaborated above. Therefore, the replacement of the existing upright SP-5 system that reached its end-of-life is critically needed.

NIH Research Projects · FY 2026 · 2025-08

Summary Intracerebral hemorrhage (ICH) poses a severe threat as the deadliest form of stroke, and effective treatments to repair the brain damage safely and permanently are yet to be established. Mesenchymal stem cell (MSC)- based therapies show promise by regulating the immune system and facilitating tissue regeneration in ICH affected brain. Recent studies underscore that the primary therapeutic benefits of MSCs come from their paracrine activity, particularly via extracellular vesicles (MSC-EVs). Unlike MSCs, MSC-EVs are much smaller (~40-200 nM) and possess the ability to breach the blood-brain barrier, delivering beneficial factors for nerve growth and immune modulation. Despite recognizing the therapeutic potential of both MSCs and MSC-EVs, understanding their unique behaviors and distribution within affected ICH regions remains incomplete. Moreover, once delivered safe and sensitive methods for precise tracking cells/EVs in vivo, are lacking. So far, the clinical trials for MSC therapy for ICH have been unable to draw clear conclusions about the efficacy of MSCs due to protocol inconsistencies (routes of administration, doses, frequencies, etc.). In addition, the distinct benefits of cell free (EV-based) interventions over whole cells in treating severe conditions such as ICH are not fully explored, which has further hindered advancement of cell/cell-free ICH therapies. To ensure optimal therapy, accurate mapping of the dosage based on actual biodistribution of cells/EVs in the brain and the therapeutic effectiveness of chosen administration routes is crucial. Non-invasive and quantitative monitoring of cells and EVs administered by different routes would help neurologists adjust delivery parameters and monitor immediate cell and EV dispersion to achieve optimal biodistribution profiles. Our proposed study aims to investigate Magnetic Particle Imaging (MPI) which is gaining ground as a promising 'cold' imaging technique to precisely map the distribution of Ultrasmall superparamagnetic iron oxide (USPIO) labeled MSCs and MSC-EVs in an ICH mouse model. Leveraging the high sensitivity of MPI, we seek to quantitatively assess the biodistribution of MSCs and MSC-EVs post-delivery via intravenous, intranasal, and intra-arterial routes while achieving anatomical co-registration through computed tomography (CT). Dynamic in vivo MPI of MSCs and MSC-EVs post-delivery via various routes, can provide insights into their in vivo migration (homing), report on delivery accuracy (on or off target), persistence in tissues, blood and tissue lifespan, as well as elimination pathways, all of which are critical for advancing clinically relevant cell and cell free ICH therapies in clinic.

NIH Research Projects · FY 2025 · 2025-08

T cell exhaustion is an important mechanism that controls the clinical manifestation of Type 1 (T1) diabetes in mice and humans. In humans, T cell exhaustion correlates with the length of the honeymoon phase; islets persistent in patients with a long T1D history express PDL1; and autoantibodies positive, non-diabetic patients undergoing anti-cancer treatment with PD1 inhibitors develop fulminant diabetes. In preclinical settings, PDL1 is upregulated on the β cells of NOD mice that do not develop T1D, anti-PD1 treatment induces T1D in male NOD, and transgenic expression of PDL1 on β cells prevents T1D. These data suggest that modulation of PDL1 on β cells is an important yet unexplored therapeutic opportunity to prevent diabetes clinical manifestations. We propose using a new bifunctional RNA therapeutic called PS03 to modulate PDL1 specifically on β cells. PS03 comprises the β cell-specific aptamers we have recently isolated (as targeting agents) covalently linked to a small activating RNA (saRNA) to upregulate PDL1. We hypothesize that PS03 will block diabetogenic T cell function, induce their exhaustion, and halt the autoimmune attack in T1D. We also hypothesize that PS03 will leave T1D-unrelated T cells untouched while other PD1 agonists may affect them. Our preliminary data shows that PS03, given systemically to NOD mice, PS03 upregulated PDL1 in β cells without modulating this gene in α and acinar cells nor on all other tissues evaluated. A treatment course with PS03 increased the number of pancreatic CD4+ and CD8+ T cells with an exhausted phenotype in NOD mice. A short treatment course, started one week before onset with a former unoptimized PS03 formulation, prevented T1D in 40% of the mice for over a year. Building on these promising results, we propose to compare PS03 with the soluble PD1 agonist fc-PDL1 by: 1) determining the effects of PS03-mediated PDL1 upregulation and fc-PDL1 on TCR signaling of diabetogenic and islet infiltrating T cells; 2) characterizing the effect of PS03 and fc-PDL1 treatments on diabetogenic and T1D unrelated T cells, on the immunological landscape, and β cell function; and 3) determining the effect of the two drugs on T1D progression. We will use state-of-the-art techniques such as pancreatic living slices, multiplex quantitative immunofluorescence microscopy, scRNA seq, and spectral flow cytometry to unveil the effect of PS03-induced, β cell-specific PDL1 upregulation on the immune system and insulin-producing cells. PS03 exists in both mouse and human versions; it is produced at GMP grade with an oligo synthesizer without contaminations and with a fluorinated backbone that makes it RNAse resistant and invisible to the immune system. To our knowledge, this is the only non-viral reagent that can modulate PDL1 expression selectively on murine and human β cells in vivo, allowing us to understand the mechanisms that regulate T cell exhaustion in the islets and, hopefully in the future, provide new therapeutic options to patients recently diagnosed with T1D.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY This is a K08 Mentored Clinician Scientist Development Award application for Anh Pham, MD, PhD, who is an Assistant Professor of Ophthalmology at the Bascom Palmer Eye Institute (BPEI) with a career goal of becoming an independent clinician-scientist in the field of glaucoma. Her research focus examines the metrics of mitochondrial health, including morphology, biogenesis, selective removal, and genomic stability, in the outflow facility between healthy and glaucoma patients. The research proposal seeks to identify mitochondrial dysfunction as an important mechanism for the development of ocular hypertension. The long-term goal of this project is to acquire the scientific skills needed to enhance our understanding of the regulatory mechanisms controlling mitochondrial health in the outflow facility and to develop mitochondria targeted therapies to improve intraocular pressure homeostasis. The scientific objective is to test the hypothesis that impaired mitophagy, the selective clearance of damaged mitochondria, contributes to the accumulation of mitochondrial dysfunction in glaucoma outflow facility. The project will utilize in vitro, ex vivo, and in vivo models of the outflow facility to examine the intrinsic relationship between mitochondrial health and TM physiology. There are three focused aims to test this hypothesis: 1) Compare mitochondrial function between healthy and glaucoma TM in cell culture; 2) Simulate ocular hypertension in an ex vivo model to develop a tissue platform for examining the protective effects of mitophagy induction in TM physiology; 3) Develop a mouse model with mitochondrial dysfunction in the outflow facility as a potential model for aging in the eye and ocular hypertension. Results of the proposed research will establish the foundation for an R01 application on mitochondrial quality control in the anterior segment. The career development objective is to develop the mentorship and expertise to become a productive and independent clinician-scientist. Dr. Pham has assembled a mentorship team consisting of Dr. Sanjoy K. Bhattacharya, an expert in TM biology and Dr. Carlos Moraes, an expert in in vivo models of mitochondrial dysfunction. Additionally, she has identified key collaborators and consultants including Drs. Felipe Medeiros, Richard K. Lee, Padmanabhan Pattabiraman, and Weiming Mao. She will meet regularly with her mentors and advisors to discuss career development, attend pertinent university seminars and workshops, present research annually at national conferences, consistently submit work for publication, and apply for extramural funding. The extensive clinical and scientific resources at BPEI, the world-class faculty on the mentorship and advisory committee, and the dedicated institutional commitment will provide Dr. Pham with the support needed to promote scientific independence in the study of mitochondrial health in the anterior segment.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Stroke is the fifth leading cause of death in the US but disproportionately impacts ethnic minority populations. Among the over 62 million Hispanic people in the US, stroke represents the third leading cause of death among Hispanic men and the fourth among Hispanic women. In 2023, the American Heart Association developed the Predicting Risk of CVD EVENTs (PREVENT) equations to predict risk of cardiovascular disease, including stroke as an independent outcome. The PREVENT equations are sex-specific but agnostic to race and can be used to estimate ten- and 30-year risk of stroke using both a base model and an enhanced model, which includes information on social determinants of health. Despite this flexibility, the PREVENT equations were developed using a population of primarily non-Hispanic Black and White adults. Though Hispanics constitute the largest ethnic minority population in the US, only 5% of the sample used for the development of the PREVENT equations were of Hispanic ethnicity. Therefore, performance of the PREVENT equation to estimate risk of stroke among US Hispanic adults remains unknown. Within this context, the overall objective of this NIH/NINDS R03 application is to validate the PREVENT risk equation for stroke among US Hispanic adults. To do so, we will leverage data from the Hipanic Community Health Study/Study of Latinos (HCHS/SOL). HCHS/SOL is an ongoing cohort of US Hispanics of diverse backgrounds residing in 4 cities: Bronx, NY; Chicago, IL; Miami, Fl; and San Diego, CA. We will include 4,827 male and 7,919 female participants who were ages 30 to 74 at baseline in 2008-2011 and without prevalent cardiovascular disease. The baseline HCHS/SOL visit included in person collection of clinical measures required for the calculation of stroke risk using the PREVENT equation for stroke. Participants were followed annually for ten years, and stroke events were adjudicated by a panel of expert clinicians. This framework will allow for the execution of the following aims: 1) Estimate the ten- and 30-year risk of stroke among Hispanic adults overall and by sex, Hispanic origin, and nativity, using the PREVENT equation for stroke; and 2) validate the ten-year PREVENT equation for stroke among US Hispanic adults through the assessment of calibration and discrimination. Accurate prediction of stroke risk is critical to its primary prevention. This proposal will evaluate whether the novel PREVENT risk equations for stroke accurately predict stroke among Hispanic adults.

NIH Research Projects · FY 2025 · 2025-08

Demyelinating eye diseases are a broad spectrum of pathologies affecting the optic nerve that can result in irreversible loss of visual function. The majority has an autoimmune-inflammatory etiology, but some may result from isolated inflammatory episodes or traumatic insult to the eye. Demyelination is also observed in eye diseases not classically defined as demyelinating, like glaucoma, where it contributes to visual dysfunction. The pathology typically initiates with immune-mediated oligodendrocyte (OL) cell death in the optic nerve. This leads to myelin destruction followed by Wallerian degeneration of optic nerve axons and, eventually, retinal ganglion cell (RGC) death. Available therapies are scarce and limited to immunosuppressants and anti- inflammatory drugs that only reduce symptoms and delay disease progression. Neuroprotective and neuroreparative therapies that prevent RGC degeneration and/or promote remyelination, and in turn preserve visual function, are lacking. This is largely due to poor understanding of the underlying pathological processes as well as repair mechanisms at play in these conditions. Cholesterol is vital for proper development and functioning of the nervous system. It is essential for neurite outgrowth, synaptogenesis, molecule trafficking across organelles and cells, intracellular signaling, myelination and remyelination. Given the high demand of cholesterol for nervous system health, maintenance of optimal cholesterol levels and homeostasis within the CNS is key and its disruption has been linked to neurological disease such as Alzheimer’s, multiple sclerosis (MS) and traumatic brain injury (TBI). Importantly, alterations of cholesterol homeostasis and metabolism have been described in demyelinating eye diseases, where RGCs and OLs, which heavily rely on cholesterol for survival, axonal transmission, myelin formation and repair, are disproportionally affected by damage. Remarkably, little has been done to address how maintenance of cholesterol homeostasis within the visual system may favor neuroprotection and repair in demyelinating eye disease, and filling this gap in knowledge may direct towards much needed new therapies. Our overarching hypothesis is that restoring cholesterol homeostasis is therapeutic in demyelinating eye disease. This will sustain neuroprotective and reparative remyelination mechanisms, whose failure is directly responsible for irreversible loss of visual function in these pathologies. To investigate our hypothesis, we propose two aims that will explore, with the experimental optic neuritis model in mice, two approaches to modulate cholesterol homeostasis: 1) administration of the cholesterol carriers methyl-β-cyclodextrins (mβCDs) to redistribute cholesterol within the demyelinated eye and provide it where needed for cell survival ad repair; 2) inhibition of the sterol sensor TMEM97, both pharmacologically and with gene targeting strategies, in order to favor neuroprotection and neurorepair.

NIH Research Projects · FY 2026 · 2025-04

Project Summary. Despite advances in immunotherapies, understanding the metabolic needs of our adaptive immune system is crucial for effective tumor control. When T lymphocytes encounter tumor antigens, they transition from a naive to an activated state. This activation necessitates a coordinated initiation of the mitochondrial biogenesis program, essential for anabolic growth and anti-tumor activity. However, the exact mechanism by which the T-cell receptor regulates this mitochondrial program remains unclear. To identify genetic regulators of mitochondrial biogenesis, I conducted a hierarchical clustering analysis to uncover patterns of gene expression during the early stages of T-cell activation. Intriguingly, this analysis revealed that the canonical pathway known for orchestrating mitochondrial biogenesis seen in oxidative tissues is absent in T cells. Instead, there was a marked upregulation of a gene called PPARG Related Coactivator 1 (PPRC1). The biological function of PPRC1 and its tissue-specific roles in mitochondrial biology are poorly understood, largely because mice with a whole-body knockout of the Pprc1 allele die post-implantation. Human PPRC1 has low sequence homology (only 12.4%) with PGC-1α and lacks the transcriptional repression motif, suggesting a distinct mode of action. Given these findings and their implications for T-cell immunity, I hypothesize that PPRC1 plays an essential role in driving a unique transcriptional program necessary for mitochondrial biogenesis to regulate T cell differentiation and anti-tumor function. To test this hypothesis, we have generated T cell-specific Pprc1 KO mouse models. In Aim 1 of the proposal, we will examine the fundamental yet largely unknown role of PPRC1 in regulating mitochondrial mass and metabolism in T cells. This aim will identify the transcriptional targets of PPRC1 and elucidate how early TCR ligation induces PPRC1 expression. Aim 2 will define the role of PPRC1 in T cell development, maintenance, and anti-tumor immunity in vivo. Overall, these studies have profound implications in cancer immunology and will challenge the existing paradigm of the mitochondrial biogenesis program during T cell activation. They promise to yield novel approaches to modulate mitochondrial function to enhance anti-tumor immunity. Candidate. Dr. Kiran Kurmi, Ph.D., is the Principal Investigator for this research proposal. As a postdoctoral fellow at Harvard Medical School, he developed tools to study mitochondrial biology and the metabolic needs of T cells. He has secured a tenure-track position for the fall of 2024 and has mapped out a detailed professional development plan to facilitate his transition to an independent researcher. His rigorous training in cancer biology and immunology, combined with his expertise in mass spectrometry and bioinformatics, uniquely positions him to establish an innovative research program examining how mitochondrial mechanisms influence T cell functions and tumor growth. In the long term, he aims to build a solid framework to explore the prevalence of novel metabolite-protein interactions in T cell biology, marking a research path distinct from that of his past mentors.

NIH Research Projects · FY 2026 · 2025-04

ABSTRACT For intermediate and high risk prostate cancer patients who are candidates for radiotherapy (RT), the only recognized early clinical response indicator is serum PSA level. There are some risk features used for treatment intensification, but optimized criteria are lacking for recommendations concerning the use and length of androgen deprivation therapy (ADT), and the use of additional systemic agents (e.g., abiraterone, chemotherapy). Approximately 30-40% of patients diagnosed with unfavorable intermediate to high risk nonmetastatic prostate cancer will develop biochemical or clinical progression within 5 years and about half of men with distant metastasis from prostate cancer originated from these groups. There is a critical unmet need to better define risk of progression such that treatment intensity is optimized and progression prevented. We propose to define the role of quantitative multiparametric MRI (mpMRI) features in combination with Circulating Tumor Cells (CTCs) as early markers of patient outcome after RT ± ADT in unique longitudinal clinical studies strategically carried out at the University of Miami and in an external radiotherapy validation cohort from the NCI. Our proposal addresses the problem of prostate tumor heterogeneity by applying an mpMRI 10 point pixel-by-pixel automated habitat risk score (HRS) classification system based on spatially distinct tumor/microenvironment characteristics. The habitat concept was introduced to map and adjust for tumor heterogeneity. The established “absolute” scale of HRS is ideal for monitoring changes longitudinally while reducing MRI vendor effects. In addition, we will apply more detailed quantitative features in models that link the changes in habitat composition and radiomics to liquid biopsy information and clinical outcome measures. The primary early endpoint is freedom from biochemical nadir thresholds at 9 months (FFBN9mo). The secondary longer term endpoint is time to Biochemical and/or Clinical Disease Failure (BCDF). Our group has recognized that liquid biopsy measures have the potential to improve the appraisal of pretreatment risk and longitudinal response to treatment. As such, CTCs, quantified using a microfiltration system, have been measured in our clinical trials at intervals before and after RT±ADT. In this application, we will leverage the unique clinical power of several mature and ongoing prospective RT trials in which mpMRI features (Aim 1), and CTCs (Aim 2) are collected pre-RT and at four time points post-RT. To our knowledge, these trials constitute the largest and most exhaustive resource for investigation of the relationship between quantitative imaging features, pre-Tx tissue genomics, liquid biopsies and RT treatment outcomes. The final goal of the proposal is to develop a multivariable model (Aim 3) that incorporates pretreatment and longitudinal mpMRI features and CTCs to guide physicians’ decisions about intensification and de-intensification of radiation therapy. These models will be validated in a prospective clinical trial (the Miami UAdapt trial).

NIH Research Projects · FY 2025 · 2025-03

Abstract Critical limb ischemia (CLI) is the end stage of peripheral artery disease (PAD) and can be an underlying cause of ischemic rest pain, gangrene, and amputation. Primary amputation is often required for the 30% of CLI patients who are not eligible for limb revascularization; thus, an effective therapy to improve neovascularization is urgently needed. During hindlimb ischemia, monocytes are among the first cells to home on the ischemia site and contribute to neovascularization. Our recent publication revealed that ischemia training, performed by 24 hours of unilateral femoral artery ligation, led to functional reprogramming of bone marrow-derived monocytes (BM-Mono), enabling them to protect against the outcomes of limb ischemia by increasing perfusion and neovascularization. Mechanistically, this reprogramming resulted in the downregulation of 24- Dehydrocholesterol Reductase (Dhcr24, an important enzyme that converts desmosterol into cholesterol), and a consequent accumulation of desmosterol in those cells. Interestingly, the ischemia training process led to a systemic effect that also reprogrammed BM-Mono in the contralateral limb. Here, our primary scientific goal is to expand our data to target Dhcr24 in monocytes as a novel and unique strategy to improve neovascularization in the setting of PAD/CLI. We will first identify whether extracellular vesicles (EVs) are the mechanism underlying the systemic effect of BM-Mono reprogramming; we will then investigate whether loss of Dhcr24 in myeloid cells regulates their crosstalk with endothelial cells (ECs) and consequent endothelium adhesion and transendothelial migration (TEM). We hypothesize that the reprogramming of BM-Mono occurs through systemic EVs that transfer epigenetic cargo to those cells, downregulating Dhcr24 and accumulating desmosterol. These reprogrammed BM-Mono have a lower adhesion rate to ECs and reduced TEM, reducing inflammation and leading to proper arteriogenesis. Our study has three specific aims (SA): SA1: Investigate the systemic effect of ischemia training on monocyte reprogramming We will investigate whether the cargo transfer from EVs released in the circulation during ischemia training is responsible for monocyte reprogramming. SA2: Determine whether myeloid Dhcr24 regulation modifies hindlimb ischemia outcomes We will determine the dependence of loss- or gain-of-function of monocyte Dhcr24 on limb perfusion, limb function, and arteriogenesis, using unique transgenic mouse models that overexpress or delete Dhcr24 specifically in myeloid cells; and then subject these mice to hindlimb ischemia. SA3: Demonstrate whether loss of Dhcr24 in myeloid cells regulates their crosstalk with ECs We will assess whether loss of Dhcr24 in myeloid cells regulates gene expression/phenotype of ECs and myeloid cells in ischemic-limb muscle using single-cell RNA-seq analysis. Further, we will assess if loss of Dhcr24 in monocytes reduces adhesion to the endothelium and TEM.

NIH Research Projects · FY 2024 · 2025-03

Project Summary G protein coupled receptors (GPCRs) are family of receptors that translate extracellular cues into intracellular responses by activating heterotrimeric G proteins. GPCRs are involved in many aspects of biology and play critical roles as receptors for neuromodulators, neurotransmitters, and hormones. GPCRs are characterized by their seven transmembrane domains. Despite having so much of their structure embedded in the membrane, most of the known GPCR regulators are cytosolic proteins. Thus, an unanswered question remains does the membrane shape GPCR signaling and is the cell able to control the local membrane landscape to modulate GPCR function. The mu-opioid receptor (µOR) is a prototypical class A GPCR with clinical relevance as it is the molecular target of opioids and drives the analgesic effect of these drugs, as well as the undesirable side effects that make the ongoing opioid abuse epidemic so devastating. In a screen for µOR regulators, we identified the cholesterol binding protein patched domain containing-1 (PTCHD1) as a negative regulator of opioid action. PTCHD1 is a critical regulator of opioid action and its knockout in mice results in elevated opioid efficacy without the development of tolerance. Our data suggests that PTCHD1 acts to shape the local membrane environment reducing the accessibility of cholesterol. Here we explore how PTCHD1 and membrane cholesterol regulate the µOR which will lead to a better understanding of how GPCRs are regulated by the membrane. We hypothesize that PTCHD1 acts in a localized manner to reduce cholesterol accessibility preventing membrane cholesterol from acting as a positive allosteric modulator of the µOR and altering the postendocytic trafficking of the receptor. We will test this hypothesis using a combination of cellular signaling and protein trafficking assays in cell lines as well as in primary neurons. This line of research is predicted to provide novel insights into the mechanisms that shape the action of opioids and underlie the development of tolerance, a major obstacle in the use of opioids in pain management that contributes to the opioid abuse crisis. Further, this research is poised to reveal a novel avenue of GPCR regulation through shaping the membrane lipid composition.

NIH Research Projects · FY 2026 · 2025-03

In this proposal we aim to identify epigenetic dependencies in breast cancer to enhance the survival rate of patients. We aim to investigate epigenetic vulnerabilities in endocrine-resistant breast cancer, specifically targeting the newly discovered CoREST-SWI/SNF axis. Around 80% of breast cancer cases are estrogen receptor positive (ER+). Existing treatments target the estrogen pathway, but approximately 40% of patients develop intrinsic resistance, leading to recurrence within 5 to 10 years. Furthermore, acquired antiestrogen resistance is common in the metastatic setting, contributing to patient mortality. Therefore, there is an urgent need to identify new therapeutic targets and mechanisms to combat metastatic endocrine-resistant ER+ breast cancer. Our study focuses on identifying epigenetic mechanisms mediated by a CoREST-SWI/SNF axis in two key resistant settings of endocrine resistance. We recently discovered that the CoREST complex, which includes the histone deacetylases HDAC1/2 and histone H4 lysine 4 demethylase LSD1, regulates breast cancer cell plasticity and endocrine therapy resistance, mediated by the loss of ESR1 expression (which encodes for ER). Additionally, we showed that CoREST interacts with the ATPase chromatin remodeling SWI/SNF complexes, and genetic and pharmacological inhibition of CoREST block tumorigenesis of endocrine resistant cells. Mechanistically, CoREST-SWI/SNF regulates the ER pathway in endocrine sensitive cells while regulating the AP-1 pathway in endocrine resistant cells upon loss of ER. Given our discovery of the functional crosstalk between ER pathway and CoREST and physical interaction between CoREST and SWI/SNF, here we propose two specific aims to dissect the CoREST-SWI/SNF axis endocrine resistant breast cancer mediated by acquired mutations in the estrogen pathway and SWI/SNF. In aim 1, we will determine the role of the CoREST-SWI/SNF axis in endocrine-resistant breast cancer with ESR1 mutations. We will study the role of CoREST in cell proliferation, its binding to chromatin in the presence of ESR1 mutations, and its physical and functional association with SWI/SNF both in cell lines and in xenografts and PDX models. We will also evaluate the effects of CoREST inhibition, alone or in combination with SWI/SNF inhibition on tumor burden. In aim 2, we will determine the role of CoREST in endocrine-resistant breast cancer with mutations in the SWI/SNF gene ARID1A. To this end, we will test whether CoREST inhibition renders ARID1A-deficient cells sensitive to fulvestrant, both in vitro and in vivo. Additionally, we will mechanisms of chromatin accessibility, gene regulation and tumor burden mediated by CoREST in cells lacking ARID1A. This proposal aims to improve the overall survival of patients with metastatic breast cancer by identifying novel therapeutic targets and mechanisms.

NIH Research Projects · FY 2026 · 2025-02

Posterior uveitis accounts for up to 15% of severe visual impairments in the United States. Mostly affecting individuals in childhood or young adulthood, uveitis is comparable to diabetes and macular degeneration in terms of years of visual morbidity. The disease etiology is largely unknown. Experimental autoimmune uveitis (EAU) is an animal disease that shares essential pathological features with human uveitis and is a valuable model for studying mechanisms of human uveitis and for evaluating efficacy of new therapies and diagnostic methodologies. However, a significant limitation with the experimental models including EAU is that assessment and evaluation of the disease and its severity is subject to substantial inter- and intra-observer variability and results from lab to lab are rarely reproducible. This shortcoming directly impacts quantification and accurate assessment of disease, which in turn is crucial for assessing new medical therapies. Artificial Intelligence (AI) has shown promising applications in ophthalmology and many deep learning models have been successfully applied to the screening and diagnosis of a growing list of clinical ocular conditions. Yet, AI models, such as deep learning algorithms, have not been extensively studied in experimental research such as EAU. We hypothesize that emerging foundation AI models based on self-supervised learning and vision transformer units will be able to provide a robust generalizable platform for experimental research thus improve the objectivity and reproducibility as well as accuracy of quantification and disease assessment. The overarching aim of this study is to provide AI platforms to unify experimental disease assessment with an application on EAU. We propose to 1) generate high-quality and well-annotated multi-modal datasets from three independent centers with fundus, optical coherence tomography (OCT), histological, and immunohistochemical images from EAU models and 2) develop a foundation model based on emerging AI models to quantify and assess uveitis images with potential generalization to other experimental diseases. We will validate the AI models using independent subsets of images to assure reproducibility and generalizability. To achieve these objectives, we have assembled a team of experts with integrated and complementary skills in uveitis, imaging, and AI with access to large multi-modal uveitis datasets generated previously at NIH and two other institutes in New Zealand and UK. We will publish datasets at TVST or Ophthalmology Science journals and make datasets and annotations publicly accessible to the community. We will also publish AI tools and make them publicly accessible to the vision community based on the NEI guidelines.

NIH Research Projects · FY 2026 · 2025-02

ABSTRACT In the era of successful combination antiretroviral therapy, human immunodeficiency virus (HIV) infection has shifted from a rapidly deteriorating disease to a manageable chronic condition with life expectancy similar to that of individuals without HIV. However, the prolonged lifespan has brought about an increase in comorbidities such as chronic obstructive pulmonary disease, type 2 diabetes, cardiovascular disease, and sudden cardiac death (SCD). Sleep disorders, particularly obstructive sleep apnea (OSA), are prevalent among those living with HIV, with recent data indicating a 47% higher prevalence in men with HIV compared to those without. OSA, characterized by upper airway collapse during sleep, is known to independently increase the risk of hypertension, cardiovascular disease, and cardiac arrhythmias, including SCD. Ventricular repolarization, a crucial aspect of cardiac electrophysiology, is destabilized by OSA-induced sympathetic nervous system activation, potentially leading to life-threatening arrhythmias. The QT variability index (QTVI), reflecting beat-to-beat QT interval fluctuations derived from the surface electrocardiogram (ECG), is a predictor of ventricular arrhythmias and SCD. Our preliminary data suggest that prevalent OSA is associated with increased ventricular repolarization lability (increased QTVI), and incident OSA exacerbates this lability. Given our previous findings from the Multicenter AIDS Cohort Study (MACS) of an increased QTVI in men with than without HIV, we aim to investigate the interaction between OSA and HIV on ventricular repolarization abnormalities. Our overarching goal is to delineate the independent and interactive effects of OSA and HIV on QTVI during sleep in a cohort of 851 men living with and without HIV. We hypothesize that OSA severity, measured by the apnea-hypopnea index (AHI), degree of nocturnal hypoxemia, and frequency of arousals, will be independently associated with higher QTVI. Furthermore, men with HIV are expected to exhibit higher QTVI than those without HIV, with OSA exacerbating the HIV-related increase in QTVI. Utilizing polysomnographic and ECG data from the MACS, we will employ multivariable regression models adjusted for relevant covariates to explore these associations. Anticipated results include independent associations between OSA severity, measures of hypoxemia and arousal frequency, and ventricular repolarization lability, along with a heightened repolarization lability in men with HIV. We expect OSA to augment the effects of HIV on repolarization lability, potentially increasing SCD risk. Given that OSA is pervasive in the HIV population, findings from the proposed analyses will have a fundamental impact on clinical practice by: (a) providing much-needed information on the pathogenesis of SCD risk in HIV, and (b) promoting early identification and treatment of OSA in those with HIV.

NIH Research Projects · FY 2025 · 2025-02

Project Summary - Abstract Long-term delivery of potent broadly neutralizing antibodies (bnAbs) using adeno-associated virus (AAV)- mediated gene transfer represents a promising approach for the prevention and treatment of HIV infection. However, the effectiveness of AAV-antibody gene transfer has been hindered by the emergence of unwanted immune responses including anti-drug antibodies (ADAs), which have been observed in both monkeys and humans. Here we present strategies that aim at preventing or eliminating host immune responses toward AAV- delivered bnAbs. A wide array of immunomodulatory drugs has been successfully used in the fields of autoimmunity, transplantation, and cancer medicine. Therefore, a well-defined drug combination regimen could be employed to increase the efficacy of AAV gene therapy. The experimental project will enroll ten naive AAV- negative rhesus monkeys that will be segregated into two groups. In our first aim, we will attempt to induce tolerance toward AAV-delivered antibodies by a synergistically acting drug treatment prior to, during, and following AAV administration. In group 1, five rhesus monkeys will receive intramuscular injections of rapamycin and ibrutinib three times a week for the duration of ten weeks. At week 2 following the initiation of the drug treatment, the monkeys will be given AAV9 encoding the C-rh 3BNC117 anti-HIV bnAb. The five rhesus monkeys in group 2 will serve as controls and will only be given AAV9 encoding C-rh 3BNC117 in the absence of any drug treatment. Levels of AAV-delivered antibody and ADAs will be monitored in serum and compared among the two groups through week 26. In our second aim, we will explore the use of nucleoside-modified mRNA delivery for its potential to induce antigen-specific tolerance. In mouse experiments, this approach has been demonstrated to prevent de novo immune responses against a foreign protein and to have immunomodulatory properties when used as treatment of autoimmune disease (Krienke et al., Science 371, 145–153, 2021). At week 26, nucleoside- modified mRNA lipid nanoparticles encoding the C-rh PGT145 bnAb will be given to the five control animals of group 2 with the goal to preventing de novo immune responses toward AAV8-delivered C-rh PGT145 that will be administered at week 32. At week 26, the animals of group 2 will also receive nucleoside-modified mRNA lipid nanoparticles encoding the C-rh 3BNC117 bnAb with the goal to reverting expected ADAs toward the previously AAV9-delivered C-rh 3BNC117. Serum levels of C-rh PGT145 and C-rh 3BNC117 and ADAs will be monitored through week 48 and the data will be compared among the two groups and against historical control animals that consistently elicited ADAs toward the two bnAbs following AAV administration. These experiments will define which of several conditions will, or will not, result in consistent tolerance to AAV-delivered anti-HIV antibodies. The use of synergistically acting drugs and immunomodulatory mRNA intervention hold promise to solve the ADA problem with the ultimate goal of preventing and eliminating HIV infection in the human population.

NIH Research Projects · FY 2026 · 2025-02

Immunotherapy has revolutionized cancer treatment and has become the fourth pillar of cancer care alongside surgery, radiation, and chemotherapy. Immune checkpoint inhibitors work by binding to proteins in order to enable the immune system to recognize and attack cancer cells. Although checkpoint inhibitors offer patients with metastatic solid tumors new options for effective, life-prolonging care, treatment can be accompanied by a host of mild to moderate toxicities and, in some cases, severe or life- threatening immunotherapy-related adverse events (irAES) that cause significant decrements to health-related quality of life (HRQoL). Toxicities vary by ICI class, up to 95% of patients experience at least one toxicity of any grade and between 10% to 47% experience at least one severe (grade 3 or 4) toxicity. Serious irAES like hepatitis, pneumonitis, colitis, and other autoimmune reactions may require immediate management or hospitalization. Toxicities may occur shortly after treatment or emerge as late effects months after treatment. Despite the serious and debilitating consequences of clinician-reported toxicities, limited work has captured patients’ experience on checkpoint inhibitors using patient-reported outcomes (PROs) or identified multilevel predictors and clinical endpoints of symptomatic toxicities and HRQoL. Therefore, the aims of this study are to: Aim 1a) characterize toxicities and HRQoL over time among individuals receiving checkpoint inhibitors for metastatic cancer (N=416) and Aim1b) examine associations with sociodemographic and medical characteristics to identify patient profiles who are at elevated risk for toxicities and decrements in HRQoL; Aim 2) examine potentially modifiable behavioral, psychosocial, and care delivery factors that account for associations between sociodemographic/medical characteristics and toxicities and HRQoL to inform targets of future intervention; Aim 3) examine whether toxicities and HRQoL predict clinical endpoints, including ICI modification, discontinuation, or reinitiation, clinician-reported irAES, cancer progression, death, and healthcare utilization (hospitalization, ED visits, use of oncology nurse triage line). Furthermore, we will use cutting-edge machine learning methods as the basis to create a nomogram prediction tool– the first of its kind that (with future validation) will allow clinicians and patients to estimate a patient’s risk of toxicities and related endpoints using PROs. With our history of successful collaboration and robust immunotherapy programs, the University of Miami Sylvester Comprehensive Cancer Center and UT Health San Antonio Mays Cancer Center are uniquely positioned to conduct this research. In alignment with NCI Moonshot Initiative priorities, this research will generate crucial knowledge to inform intervention efforts to improve the safety and tolerability of checkpoint inhibitors and prevent and mitigate toxicities among patients.

NIH Research Projects · FY 2025 · 2025-01

Abstract Hearing loss (HL), congenital or acquired, is a major health issue which affects approximately 30 million people in the United States alone. Genetic causes are one of the most common etiologies. So far, >200 genes have been identified as causative for HL in humans. This work is significant because we propose to merge two approaches to develop a platform by generation of human inner ear organoids (hIEOs) from human pluripotent stem cells (hiPSCs) as a model system to test inner ear gene therapies and introduction of advanced genome editing strategies to silence dominant mutations or repair recessive mutations that cause dysfunction in human sensory hair cells. To support the role of identified variants in pathophysiology, we will undertake novel in vitro functional studies of these IEOs. The proposed human studies are complementary with development of a human iPSC-derived model for human audio vestibular organoids and analysis of the effect of HL mutations on IEOs. No single system or experiment is typically conclusive in itself. Therefore, to support the role of identified variants in pathophysiology, we will undertake novel in vitro functional studies of these IEOs including using novel muti-`Omic approaches including scATAC-seq & scRNA-seq, epigenetic, and organoid-on-a-chip' platforms. The short-term impact of this proposal will be to generate a novel in vitro human iPSC-derived model corresponding to the vestibular region affected in hearing and balance and to evaluate the cellular consequences of the CRISPR/Cas9-based exon- skipping or disruption strategies in human cells. The long-term impact will be to establish a new paradigm for personalized genetic models of hIEOs affected in HL for in vitro screening of available therapeutics and multi-omic identification of disease and biomarkers for treatment responses. We hypothesize that human induced pluripotent stem cells (iPSCs) and IEOs provide valuable experimental platforms for testing potential therapeutic strategies and gene targeting for the correction of audiovestibular disorders. We further hypothesize that in vitro CRISPR/Cas9-based gene disruption strategies can rescue HL-associated mutations in human hair cells (HCs) using human IEOs laying the foundation for a human clinical trial in the treatment of HL. Specific aims of the project are: 1 Establishment and optimization of hiPSC-derived 3D inner ear system from patient-with genetic HL. 2.Test in vitro CRISPR/Cas9-based gene disruption and AAV-based gene expression strategies to model HL-associated IEO cell defects and to rescue phenotypes-associated mutations in the human IEOs. These innovative approaches for treatment of genetic HL will pave the way for translation to clinical trials. We are optimistic that by merging these technologies we can successfully translate these innovations for other inner ear disorders and, importantly, broader use in other genetic disorders.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY The parasympathetic nervous system uses the vagus nerve to modulate the secretion of both insulin and digestive enzymes from the pancreas. Because of its diffuse innervation pattern targeting multiple visceral organs as well as the indirect approaches used to assess the role of the vagus nerve in the pancreas, interpretation of the existing evidence is difficult. Furthermore, intrapancreatic neurons, which are the exclusive targets and mandatory relays of the efferent vagus nerve in the pancreas, have not been sufficiently studied. It is widely believed that they act as mere conduits of vagal input and do not substantively contribute to autonomic control of pancreas function. In preliminary studies, I found that intrapancreatic neurons respond to cholinergic input through stimulation of both nicotinic (nAChRs) and muscarinic acetylcholine receptors (mAChRs). Neuronal responses elicited by mAChR activation, which do not occur in canonical parasympathetic ganglia, resulted in long lasting, rhythmic bursts of activity. Additionally, I found that intrapancreatic neurons possess the molecular and functional signatures of the neuronal M-current, a voltage gated K+ conductance mediated by Kv7.2/7.3 ion channels indirectly inhibited my mAChR activation. These results indicate that intrapancreatic neurons indeed possess functional characteristics supporting the notion that they are centers integrating and processing cholinergic input from the vagus nerve. I therefore hypothesize that intrapancreatic neurons transform cholinergic input through an mAChR associated mechanism and rely on the function of Kv7.2/7.3 ion channels to achieve efficient control of pancreatic targets. In Aim1, I propose to use living pancreas slices generated from a transgenic mouse model that expresses the genetically encoded Ca2+ indicator GCaMP3 in intrapancreatic neurons to understand the contribution of mAChR activation and associated M-Current inhibition on intrapancreatic neuron activity. I will also use electrophysiologic techniques to interrogate the M-Current. In Aim2, I will explore how Kv7.2/7.3 channels influence the transmission of neuronal input from intrapancreatic neurons to exocrine and endocrine targets within the pancreas following targeted neuronal chemogenetic stimulation. The rationale for the proposed research is that, if we want to develop therapies leveraging electrical stimulation of the vagus nerve to treat or prevent diseases like diabetes and pancreatitis, it is imperative to elucidate the mechanisms by which intrapancreatic neurons, not only relay, but integrate and transform vagal cholinergic input to the pancreas. Additionally, Kv7.2/7.3 channelopathies due to mutations in KCNQ2 and KCNQ3 can result in severe epilepsy and developmental delay as well as gastrointestinal symptoms and autonomic dysfunction. The proposed research is significant because it will produce mechanistic insight into how local intrapancreatic neurons functionally impact endocrine and exocrine compartments of the pancreas and will lay the groundwork for studying the impact of Kv7.2/7.3 channelopathies on pancreas function.