Methodist Hospital Research Institute

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract: This K08 proposal outlines a five-year research and training plan that will propel Dr. Fan’s career as an independent physician scientist and expert in microbiome manipulations in immunotherapy enterocolitis (IMEC). Immune checkpoint inhibitors have revolutionized cancer treatment by leveraging our own immune system against tumor cells. Unfortunately, off-target effects, including inflammation of the gastrointestinal tract, like IMEC, can occur. Antibiotic exposure leads to dysbiosis which disrupts the careful interplay between beneficial and harmful bacteria. Antibiotics, specifically those with anaerobic spectrum increase the incidence of moderate-severe IMEC. In cancer patients, antibiotic exposure often leads to gut microbiome domination with vancomycin-resistant Enterococcus faecium (Efm), and exposure to anaerobic spectrum antibiotics can also target beneficial members of the Bacteroides spp., which play a critical role in carbohydrate degradation. Indeed, Bacteroides spp. have a diverse repertoire of carbohydrate degrading enzymes with gene groups predicted to code for Carbohydrate-Active enZyme (CAZyme) families, designated polysaccharide utilization loci (PUL), which coordinate complex carbohydrate degradation. Our preliminary patient data show an expansion of B. xylanisolvens in IMEC patients who responded to fecal microbiota transplantation (FMT). Moreover, CAZyme genes (encoding GH16 and GH3) increase after FMT in responders, which can be attributed to B. xylanisolvens. Further, these enzymes degrade the prebiotic product, beta-glucan. On the other hand, clade A Efm has been associated with hospitalized and critically ill patients who have received massive amounts of antibiotics and isolates belonging to this clade have been shown to worsen colitis, in genetically susceptible il10-/- mice. Antibiotic exposure and the gene abundance of a “resistant allele” of pbp5, coding for penicillin-binding protein 5 (PBP-5) and mediating ampicillin resistance in Efm is typical of clade A. Our data suggests that enterococci are more abundant in patients with IMEC and enterococcal abundance decreases in responders after FMT. This balance of good and bad bacteria may be critical to IMEC development. Developing microbiome targeted opportunities to mitigate disease severity and activity is promising. Based on preliminary data from our upfront FMT cohort, we find that two specific bacteria that are associated with antibiotic exposure may play a role in mediating severity in IMEC. Specifically, we aim to investigate the contributions of the following in the development and severity of IMEC: 1) carbohydrate degradation of B. xylanisolvens and prebiotic, beta-glucan and 2) antibiotic exposure and clade A Efm predominance. This K08 award will facilitate the mentorship and career development of Dr. Fan with world experts to help elucidate the role of these two bacteria in IMEC development. With mentors from Houston Methodist and the Texas Medical Center, led by Dr. Cesar A Arias, MD, PhD., discoveries from this award will set the basis for an R01 on the mechanisms of both Efm gut colonization and the functional role of carbohydrate degradation by B. xylanisolvens in IMEC.

NIH Research Projects · FY 2026 · 2026-06

Contact PD/PI: MYLONAKIS, ELEFTHERIOS PROJECT SUMMARY: Disruptions in microbial homeostasis due to age, antibiotic use, chronic illness, or immunosuppression promote overgrowth of opportunistic pathogens such as Candida albicans and Streptococcus mutans. This contributes to oral conditions like candidiasis, caries, and mucosal inflammation, and is linked to cardiovascular disease, diabetes, autoimmune disorders, adverse pregnancy outcomes, and gut-brain axis disturbances. Conventional antifungal therapies often fail to resolve biofilm-associated infections and may drive resistance, underscoring the need for safe, locally-acting alternatives. We have developed Lactobacillus paracasei 28.4-based probiotic strategies to restore mucosal homeostasis and target fungal and bacterial pathogens. Originally isolated from a caries-free individual, L. paracasei 28.4 inhibits C. albicans and S. mutans by reducing adhesion, biofilm formation, and virulence gene expression. Delivered via gellan gum hydrogels, it significantly lowers fungal burden and inflammation in murine oropharyngeal candidiasis models. Its postbiotic components also suppress Candida auris and drug-tolerant persister cells. Preliminary studies further enhanced efficacy by integrating L. paracasei 28.4 with caffeic acid phenethyl ester (CAPE), a natural antifungal and immunomodulator, in biocompatible hydrogels enabling controlled release and targeted mucosal delivery. We propose to advance gellan-based hydrogels with L. paracasei 28.4, with or without CAPE, to inhibit virulence, disrupt C. albicans and S. mutans biofilms, enhance epithelial barrier function, and modulate innate immunity. Aim 1 will optimize formulation for stability, probiotic viability, controlled release, and epithelial compatibility. Aim 2 will test safety, microbiome effects, and efficacy in murine models of candidiasis and dysbiosis. Aim 3 focuses on translational development, including in vitro safety testing, evaluation in immunocompromised hosts, and GMP-aligned formulation. This project introduces a novel, multi-modal strategy to manage oral dysbiosis via localized delivery of probiotics and natural antifungals—limiting pathogen overgrowth, restoring microbial balance, and reducing reliance on conventional drugs. Our gellan-based hydrogel co-delivers L. paracasei 28.4 ± CAPE directly to oral tissues, disrupting C. albicans and S. mutans biofilms, preventing recolonization, and preserving mucosal integrity. By maintaining probiotic viability and enabling targeted CAPE release, the system achieves strong efficacy in murine models. This biocompatible platform integrates pathogen control, immune modulation, and microbiome restoration, offering a transformative, resistance-limiting approach for difficult oral infections. References Cited Page 1

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Age-related macular degeneration (AMD) is the leading cause of irreversible blindness involving gradual dysfunction and degeneration of photoreceptors and the retinal pigment epithelium (RPE). Mitochondrial dysfunction represents a hallmark of AMD. Excessive and persistent reactive oxygen species (ROS) damages mitochondrial DNA (mtDNA), resulting in fewer mitochondria. Impairments in respiratory chain activity and reduced oxidative phosphorylation (OXPHOS) impacts ATP synthesis and further drives ROS production. Our prior work has shown that exogenous mitochondrial transplantation to cells can induce a bioenergetic shift towards increased OXPHOS and ATP production, and reduced ROS. Mitochondrial cell-to-cell transfer in response to stress rescues aerobic respiration to reduce deleterious cell dynamics. We aim to create a sustainable source of mitochondria proximal to the RPE in the form of mesenchymal stem cells (MSCs) transfected with nuclear respiratory factor 1 (NRF1), a driver of mitochondrial biogenesis, for enhanced cell-to- cell transfer of mitochondria. Our objectives are to determine how NRF1 transfection changes MSC transcriptional activity, mitochondrial production and trafficking, and extracellular vesicle (EV) contents, and test the strategy in two rodent models of retinal degeneration. We hypothesize that our cell therapy using NRF1- overexpressing MSCs will not only provide the needed energetic boost via mitochondrial mass transfer to retinal cells, but also induce profound cellular reprogramming through the secretion of EV-containing factors, thereby counteracting and potentially reversing the onset of late-stage dry AMD. We will use a combination of molecular and biochemical methods, including transcriptomics, to characterize the potency of mitochondrial transfer to impact AMD. We will first investigate the impact of NRF1 on mitochondrial transfer mechanisms in MSCs. We will elucidate the role of NRF1 on mitochondrial transfer mechanisms, specifically tunneling nanotubes (TNTs) and EVs, between MSCs and human RPE cells from donor eyes afflicted with AMD, shedding light on NRF1- primed MSCs as mitochondria generators, responsive to signals from stressed cells. We will then examine the influence of NRF1 on MSC senescence and EV composition, providing insights into the multifaceted benefits of NRF1 in MSCs, including metabolic adaptation to AMD stress environments and modulation of EV constituents. Lastly, we will evaluate the impact of NRF1 primed MSC transplantation in vivo. Through longitudinal and long- term efficacy studies and tissue transcriptomic analysis, we will identify disease stages most responsive to NRF1 MSCs when transplanted subretinally in a sodium iodate (NaIO3)-induced model of retinal degeneration and senescence accelerated mouse (SAMP8) model, highlighting the potential to rescue RPE cell degeneration and abrogate AMD progression. Findings from this work will provide crucial insights into the potential of NRF1-based therapies for AMD, offering a promising avenue for the treatment and management of this debilitating disease.

NIH Research Projects · FY 2026 · 2026-04

Project Summary The Gasdermin (Gsdm) family consists of six paralogous genes encoding GSDMA, GSDMB, GSDMC, GSDMD, GSDME and F (GSDMF also known as PJVK or DFNB59). Gsdm proteins play a crucial role in innate immunity, particularly in inflammation and the initiation of pyroptosis, a form of programmed necrotic cell death. Unlike many other immune modulators, transcriptional regulation of Gsdm family proteins is not sufficient to execute their biological functions. Gsdms A-E share highly conserved N-terminal (N-ter) and C-terminal (C-ter) domains separated by a variable linker. The C-term exhibits self-inhibition by completely masking the N-ter hydrophobic pocket that binds lipids. Thus, even if upregulated transcriptionally, Gsdm cannot be functional until it is cleaved by a protease to release the N-ter from the self-inhibited C-term. Thus, the key for understanding Gsdm biology is the identification of the protease. This work is based on our robust unpublished data where we identified a new protease that can directly cleave both human and murine GSDMC. Rooting from this discovery, we further identified three unique features of cleaved and activated GSDMCN-ter not observed in other Gsdm family members in terms of its molecular properties, cellular functions and in vivo phenotypes. (1) In contrast to other protease-processed Gsdm family members, such as GSDM-A, -B, -D and -E, the N-terminus of GSDMC (GSDMCN-ter), processed by the newly identified protease, did NOT effectively promote pyroptosis. (2) This is because GSDMCN-ter does not localize to the plasma membrane but to other subcellular organelles. (3) GSDMCN-ter has an immune regulatory role in animal models via amplifying type-2 immune responses, unrelated to pore formation at the plasma membrane (e.g. IL-1 family member cytokine release) and pyroptosis. Accordingly, we have planned three aims to understand the mechanisms controlling these features. In Aim 1, we will analyze the non-conserved amino acids between GSDMCN-ter and other Gsdms proteins that explain the pyroptosis deficiency. We will further determine the lipid binding profile of GSDMCN-ter and try to understand how it differs from that of other Gsdms proteins. In Aim 2, we will systematically investigate the intracellular membrane structures that can be targeted by GSDMCN- ter and assess the biological consequences after GSDMCN-ter targeting. Last, in Aim 3 we will use two animal models to determine how GSDMCN-ter amplifies type 2 immune responses. In summary, Aim 1 and 2 will unveil novel mechanisms of how GSDMC functions different from other Gsdms, as well as its new roles in cell biology (not effectively promoting pyroptosis). For Aim 3, with our unique animal models, we will reveal new mechanisms of actions for GSDMC as well as therapeutic strategies to boost type 2 immunity.

NIH Research Projects · FY 2026 · 2026-02

Summary T cells are central to transplant rejection, driving allograft destruction through differentiation into effector cells. However, the mechanisms by which effector T cells sustain persistent alloimmune responses remain unclear. Our recent studies have identified a subset of “stem-like” T cells within the alloreactive pool. These stem-like T cells possess two fundamental features: self-renewal and the capacity for continuous differentiation into effector T cells. Importantly, terminal effector T cells, despite having all the cardinal features of effector activity, rapidly undergo apoptosis and fail to sustain graft rejection in vivo. This underscores the critical role of stem-like T cells, which continuously generate effector T cells to drive allograft rejection. Understanding the fundamental mechanisms regulating T cell stemness is a key question with significant therapeutic implications. Our preliminary data reveal that T cell stemness is epigenetically regulated by enhancer of zeste homolog 2 (EZH2), the catalytic subunit of the Polycomb Repressive Complex 2 (PRC2). EZH2 functions as a histone methyltransferase, catalyzing the trimethylation of histone H3 at lysine 27 (H3K27me3), a key marker of gene repression. Deletion of Ezh2 in T cells completely abrogates their differentiation into effector cells. Furthermore, mice with T cell-specific EZH2 deletion (Ezh2fl/flCd4-Cre) or WT mice transiently treated with an EZH2 inhibitor (DZNep) accepted heart allografts long term (>100 days). These results suggest that EZH2 deletion/inhibition disrupts the stemness of alloreactive T cells, rendering them unable to sustain graft rejection. The central goal of this proposal is to elucidate how EZH2, an epigenetic repressor, regulates T cell stemness. We hypothesize that EZH2 preserves two key features of stem-like T cells: maintaining their long-term functional persistence and enabling their differentiation into effector cells. This hypothesis will be tested through two aims: Aim 1: Investigate whether EZH2-mediated repression of key transcription regulators is required for the differentiation of stem-like T cells into effector cells. Aim 2: Investigate whether EZH2 preserves T cell stemness by epigenetically repressing genes involved in apoptosis, cell cycle arrest, and functional exhaustion. Successful completion of these studies will uncover the epigenetic mechanisms governing stem-like T cell persistence and effector differentiation, providing a foundation for novel therapeutic strategies to improve transplant outcomes.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY This K76 proposal comprises a five-year research and career development plan to advance my, Dr. Hina Faisal’s, training in clinical research focused on aging and perioperative medicine, establish my independence as an investigator and prepare me for academic research leadership at the national and international level. I am an assistant professor at Houston Methodist Hospital with clinical expertise in anesthesia and critical care medicine and a research interest in delirium. Delirium is a sudden, fluctuating disturbance in consciousness and cognition and is highly prevalent in older surgical patients. With this award, I aim to develop the mastery needed in clinical aging research to develop, evaluate, implement, and distribute scalable nonpharmacological interventions capable of reducing the burden related to delirium. My long-term goal is to become a physician-scientist leader in geriatric-focused perioperative medicine. Delirium is preventable; nonpharmacological interventions, such as cognitive stimulation (CS), are the best preventive strategies . Implementing CS in acute hospital settings is a major challenge due to limited resources and low patient engagement. Virtual reality (VR) and gamification to deliver CS might be a scalable and engaging solution. Based on my preliminary work, I hypothesize that VR- based CS games will be safe, feasible, and acceptable in high-risk older patients (Aim 1). The proposed study will estimate the size of the effect of the intervention on cognitive functions, particularly sustained attention (Aim 2), and the incidence of postoperative delirium (Aim 3). To accomplish the above goals, I will conduct a randomized controlled pilot trial involving 90 patients ≥65 years old with pre-existing cognitive impairment admitted to the hospital following a major surgery. Patients will be randomized to 1:1:1 to receive VR-based CS games (intervention), watch a movie in VR (VR control), or receive standard-of-care nurse-led reality orientation (Nurse control). The trial will provide me with preliminary data to apply for an R01. To accomplish my research and career goals, my mentors and I have strategically designed a career development plan that integrates coursework, seminars, and mentorship in the following areas: (1) Incorporate geriatric medicine principles into perioperative medicine; (2) Gaining expertise in the design and conduct of clinical trials in older adults; (3) Training in neurocognitive testing and VR implementation; and (4) Enhancing leadership skills. I have assembled an outstanding group of interdisciplinary mentors and collaborators, including a leader in the assessment of health outcomes in aging (Dr. George Taffet), an international leader in technology implementation in older adults (Dr. Malaz Boustani), a clinical neuropsychologist (Dr. Kenneth Podell), and an expert in the application of VR technology (Dr. Junhyoung Kim). If successful, my research will open a new avenue of investigation in aging and delirium research, which aligns with the NIA’s strategic goal to develop effective interventions to reduce the burden of age-related disorders.

NIH Research Projects · FY 2025 · 2025-09

Abstract Breast cancer is the second leading cause of cancer-related death of women in the United States, and triple negative breast cancer (TNBC) is the most aggressive subtype. It comes back early, spreads fast, has a poor prognosis, and there are limited treatment options. Almost 90% of patients with TNBC that have spread to other parts of the body will pass away within five years, which is a lot worse compared to other subtypes, and the lungs are the most common site for TNBC to spread. Current standard therapies including platinum (Pt) drugs, taxanes, and anthracyclines are widely used but often cause severe toxicity and rapid drug resistance. TNBC responses moderately to current immunotherapies, and the overall treatment effects are heavily affected by a group of immune cells in tumor tissues, the immune-suppressing myeloid cells. This population contributes to forming immune environment which promotes tumor growth by inhibiting immune surveillance. Therefore, novel approaches are urgently needed to help these patients live longer, particularly for TNBC that spreads to distant organs such as the lungs, since they are more frequent and deadly. Reactive oxygen species (ROS) are unstable molecules within cells. While high levels of ROS can be harmful to cancer cells, they also significantly affect the behavior and interaction of cancer cells and myeloid cells in tumors. This makes them a promising target for TNBC treatment. However, most of current drugs do not produce sufficient ROS. Mild ROS not only fail to kill tumor cells but can promote tumor growth. Moreover, the immunosuppression effect remains a critical concern. It is important to design new drugs with high potency of ROS production. Accordingly, we have developed novel Pt-based therapeutics that produce robust ROS production, effectively eradicates TNBC cells rapidly, and successfully suppressed tumor growth in animal models. The goal of this project is to investigate the activities and biological mechanisms of these new drugs on both cancer and immune cells, with a special focus on TNBCs that have spread to lung. We will study the mechanisms how the drugs kill TNBC cells and assess the effectiveness when combined with current first-line TNBC treatment strategies. Moreover, we will explore their overall impact on the immune environment in lung metastatic TNBC nodules. These findings will provide a novel framework for development and assessment of drug efficacy against TNBC and other cancer types. Specific Aims: Aim 1: Elucidate how the ROS-generating Pt drug affects TNBC cells and immune-inhibiting myeloid cells. Aim 2: Assessment the performance of ROS-generating Pt drug in treating lung-spreading TNBC, along with its impacts on immune response.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Human immunodeficiency virus (HIV) is one of the top areas of the National Institutes of Health- funded nonhuman primates (NHP) research. Within this context, antiretroviral (ARVs) used in research for HIV pre-exposure prophylaxis and treatment require frequent dosing regimens. ARVs are often administered through daily oral dosing or repeated injections, which are costly as well as burdensome on animals, veterinary staff and operational resources. Moreover, inconsistency in dosing regimen or dosages could be detrimental to research outcomes. Given this, we propose the dissemination of a drug-agnostic ultra-long-acting subcutaneous drug delivery implant, the NanoDDI, as a research tool for NHP studies, through the Center for the Dissemination of Ultra-Long-Acting Antiretroviral Release Technology: DART Resource Program. The DART Program will provide the NanoDDI for use in NHP studies on HIV for long- term ARV release to replace conventional modes of administration. Sustained and constant ARV delivery from the NanoDDI is achieved through controlled drug diffusion across a nanofluidic membrane without pumps or ports. The NanoDDI technology is immediately ready for use and rapid and reproducible manufacturing permits widespread facile dissemination. The drug-agnostic technology is supported by 10 years of development and deployment in various animal models, with demonstrations of long-term sustained release of various ARVs in NHP for up to 29 months without interruption. The central tenet of the HIV-related NHP studies and alleviate animal, operational and financial burden associated with conventional administ ation methods. The aims of the proposed DART Program, which will be pursued concurrently over five DART Program is that the NanoDDI can standardize ARV delivery across wide-ranging years, are, Aim 1: Outreach and Dissemination of NanoDDI for HIV Research in NHP, Aim 2: Integration and Deployment of NanoDDI for ARV Delivery in Independent Users’ Laboratories, and Aim 3: Broaden the Spectrum of Applicability of NanoDDI within the HIV Research Community. Outreach initiatives will create awareness and promote adoption of the NanoDDI into current and future NHP HIV research. The DART Program will ensure standardized NanoDDI deployment in Users’ NHP studies for reproducible research outcomes through comprehensive training, supported by our research and veterinary staff, and Veterinary Scientific Advisory board. Concurrently, User-generated data and User-introduced novel ARVs could enhance outreach as well as broaden NanoDDI drug portfolio. Altogether, research outcomes and dataset generated inclusive of ARV pharmacokinetic and User-specific data, could be used to fill knowledge gaps, improve drug delivery and accelerate drug development in HIV research. Expected outcomes of the DART Program include: successful integration of the NanoDDI into NHP HIV studies with significant cost and burden reduction, increase research output due to cost- and time-efficient changes in ARV delivery protocols, higher quality research data due to less variability in ARV delivery, and minimize scientific and financial risks associated with NHP research.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Infectious disease remains one of the leading causes of illness and mortality worldwide. The recent coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV- 2) led to widespread illness and death, disrupting public health and economy. It has been successfully mitigated thanks to the development and global application of the novel messenger RNA (mRNA) vaccine technology. While mRNA vaccines have offered significant protection, the high mutation rate of SARS-CoV-2 necessitates ongoing development of updated vaccines to combat new variants. While mRNA vaccines have shown remarkable effectiveness against COVID-19 by stimulating both humoral and cellular immune responses, the main limitation lies in the inherent instability of mRNAs at normal conditions without protection. Therefore, current mRNA and mRNA formulations require ultralow temperatures for storage and transportation. Recognizing the instability challenge, we proposed to develop a novel “RNA-plex” technology, which uses an “carrier-base” polymer to bind to mRNA molecules, preventing them from degradation in fridge or room temperature during storage. It significantly reduces the transportation and distribution costs, and makes mRNA vaccines more accessible globally, especially in areas with limited cold chain facilities. Notably, this mRNA protection technology can universally shield various mRNAs, is compatible with multiple delivery systems, and significantly enhances mRNA translation in cells, suggesting its promising potential for diverse therapeutic and research applications. In this proposal, we will apply this mRNA stabilization technology named “RNA-plex” in the development of stable and efficient vaccines for SARS-CoV-2 prevention. Specifically, we will optimize the composition and formulation of RNA-plex to maximize its protective efficacy for SARS-CoV-2 omicron variant spike protein mRNA. Next, we will apply it in preparing prophylactic lipid nanoparticle (LNP) mRNA vaccines with high stability, enhanced antigen translation efficiency, and superior vaccination effectiveness compared to conventional approaches against emerging viral threats, using the recent SARS-CoV-2 omicron variant XBB.1.5 as a proof-of-principle model. Specific Aims: Aim 1. Stabilization of SARS-CoV-2 omicron variant mRNA using RNA-plex technology and develop an LNP mRNA vaccine. Aim 2. Evaluate the efficacy of LNP-RNA-plex vaccine in eliciting immune responses and protection against SARS-CoV-2 Omicron variant.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Mycobacterium tuberculosis (MTB), the causative agent of pulmonary and non-pulmonary tuberculosis (TB), kills nearly 1.2 million people every year, whereas ~ one-third of the global population is latently infected with Mtb. Bacillus Calmette-Guérin (BCG) is widely used as a vaccine against TB, and over one billion doses have been used so far although it only partially protects children from TB but not adults. We discovered that a novel-self adjuvanted mRNA (SamR) vaccine delivering MTB-derived antigen can protect mice against TB. In this proposal, we propose to develop new generation mRNA-based vaccines route to protect against TB using a multi-antigen-based platform which also integrates an adjuvant. A ‘systems immunology’ approach will be used to evaluate innate immunity mechanisms that control adaptive immune responses in mice vaccinated using our improved mRNA vaccines. Our goals are: Specific Aim-1: To develop a new generation mRNA-based vaccines expressing protective antigenic epitopes of Mycobacterium tuberculosis (MTB) in combination with in-built adjuvant peptides. We will develop multiple MTB protein antigen expressing mRNA vaccines that contain a unique self-adjuvant to boost immune responses. We will synthesize and evaluate more efficient linear mRNA and/or circular RNA vaccines for tuberculosis and formulate them in nanoparticles. Specific Aim-2: To characterize the molecular mechanisms of new generation mRNA vaccines. We found that our new generation SamR-vaccines induced a robust activation of innate immunity pathways (RIG-I NOD2 and TLR) in macrophages and dendritic cells. We will investigate the molecular mechanisms of protein antigen-induced Immunogenicity and C5- induced adjuvanticity using APCs from wild type C57Bl/6 mice, and transgenic mice that lack innate nodal genes. Specific Aim-3: To determine the efficacy of SamR-vaccines against tuberculosis in wild type and immunodeficient mice. We found that our novel vaccine activated trained immunity genes in macrophages. Because BCG-induced trained and adaptive immunity declines in human children by age 5 predisposing them to TB, we will investigate the boosting effect of our trained immunity inducing SamR-vaccines for protection in BCG vaccinated mice. We will use both C57BL/6 and our optimized immune competent and immunodeficient humanized model to identify parameters of protection relevant to humans.

NIH Research Projects · FY 2025 · 2025-08

Group B streptococcus (GBS) is a major human pathogen that causes significant morbidity and mortality. GBS infections are the leading cause of stillbirths, preterm births, and neonatal mortality worldwide. Owing to the lack of a human vaccine and rise in antibiotic resistance among GBS strains, the clinical utility of current GBS infection control measures is significantly compromised. Thus, new approaches and/or targets are urgently required to treat or prevent GBS infections. In the proposed study, we seek to characterize a previously unknown intercellular signaling pathway and determine its contribution to GBS pathogenesis. Recently, we discovered a broadly distributed, novel intercellular signaling pathway in firmicutes that is comprised of a new class of bacterial intercellular signals, leaderless communication peptides (LCPs), and a cognate intracellular receptor that controls virulence gene expression in concert with LCP. Without exception, the cognate intracellular LCP receptors for all the identified LCPs belong to a subfamily of the Rgg/Rap/NprR/PlcR/PrgX/AimR (RRNPPA)-super family of peptide-sensing transcription regulators, the Rgg subfamily. As a result, the possibility that LCPs act as signals for the members of other subfamilies of RRNPPA regulators is unexplored, and the potential roles of an entirely new class of LCPs in bacterial group behaviors remain under appreciated. Our preliminary data uncover a previously unknown AimR subfamily regulator and a corresponding LCP in the GBS genome that likely controls the expression of genes encoding a putative toxin and toxin-associated transporters. However, the critical knowledge regarding the components of GBS LCP signaling pathway, molecular mechanism of GBS LCP signaling, and the contribution of LCP signaling pathway and LCP-controlled putative toxin to GBS pathogenesis are yet to be elucidated. Thus, the primary objective of the proposal is to dissect the signaling mechanism of AimR-LCP pair of GBS, elucidate its regulatory activity in vivo, and determine its contribution to GBS pathogenesis. We will test the central hypothesis of the proposal that AimRgbs-LCPgbs constitute a cytosolic receptor-LCP pair that mediates intercellular signaling and controls virulence-associated traits of GBS in two specific aims. The completion of this study will provide critical insights into the role of a novel intercellular signaling pathway in the survival and virulence of a human pathogen and potentially identify novel targets for future translational strategies to treat or prevent GBS infections.

NIH Research Projects · FY 2025 · 2025-08

Project Summary This proposal addresses the significant challenge of T cell-mediated rejection in heart transplantation by investigating the role of lactate dehydrogenase A (LDHA) in T-cell effector differentiation. Effector T cells play a critical role in rejecting transplanted hearts. Our recent studies have revealed a novel framework in which naive alloreactive T cells activate, expand as effector precursor T (TEP) cells, and then differentiate into effector T cells that attack the heart allograft. This differentiation process requires precise effector gene expression, which we hypothesize is regulated by LDHA through metabolic- epigenetic pathways. LDHA is a key enzyme in aerobic glycolysis, converting pyruvate to lactate, a process more active in effector T cells compared to TEP cells. In mice with T cell-specific LDHA deletion, alloreactive TEP cells still proliferate but fail to differentiate into effector cells, resulting in heart allograft tolerance without the need for immunosuppression. This discovery highlights LDHA as a critical regulator of effector T- cell function. The proposal tests the hypothesis that LDHA regulates effector gene expression through two metabolic-epigenetic pathways: Aim 1 investigates whether LDHA increases cytosolic acetyl-CoA levels, promoting histone acetylation at effector gene loci. Aim 2 explores whether LDHA-mediated lactate production enhances histone lactylation and increases the NAD+/NADH ratio, supporting effector gene expression. Aim 3 evaluates whether inhibiting LDHA with the selective inhibitor FX11 can induce heart transplant tolerance, mimicking the effects of genetic LDHA deletion. This study will provide new insights into how LDHA regulates effector T-cell differentiation and offer innovative therapeutic strategies for improving transplant outcomes.

NIH Research Projects · FY 2025 · 2025-06

ABSTRACT Staphylococcus aureus is a major human pathogen responsible for a wide range of life-threatening infections. Many of these infections are caused by methicillin-susceptible S. aureus (MSSA). MSSA represent a major burden among S. aureus infections and are important contributors to mortality. For decades, the first line of therapy for severe MSSA infections have been the isoxazolyl-penicillins (ISP, e.g., nafcillin). However, recent data suggest that clinical outcomes in MSSA bacteremia are similar in patients treated with cefazolin (vs nafcillin), a cephalosporin with activity against MSSA that appears to be less toxic. Indeed, treatment with nafcillin seems to be associated with increased costs, more drug reactions (including hepatotoxicity, interstitial nephritis and neutropenia) and, possibly, higher mortality. Due to these concerns, an important shift in the treatment of MSSA is occurring whereby clinicians are now using cefazolin as first line of therapy for severe MSSA infections. An important concern of using cefazolin and other cephalosporins as primary therapy for these serious infections is the cefazolin inoculum effect (CzIE), defined as a cefazolin minimal inhibitory concentration of > 16 µg/ml when a high inoculum (107 CFU/ml) is used. The CzIE has been associated with failures in the treatment of deep-seated MSSA infections and with the production of certain isotypes of the staphylococcal β-lactamase. However, the characterization of the clinical impact of this phenomenon in deep-seated MSSA infections is limited. In addition, it is currently not possible to detect the CzIE in a standard clinical microbiology laboratory given the cumbersome and expensive nature of the gold standard test for its detection. Our published and preliminary clinical data suggest that the CzIE is an important contributor to worse clinical outcomes of severe MSSA infections. Furthermore, we have developed and published a novel colorimetric nitrocefin-based rapid test (~3 h) that detects the CzIE with high sensitivity and specificity that can be incorporated in the routine clinical microbiology laboratory. We postulate that, i) the CzIE negatively impacts clinical outcomes in MSSA bacteremia treated with cefazolin and, ii) a rapid test can be readily implemented for the identification of the CzIE in S. aureus bacteremia and can detect patients at higher risk of poor outcomes. In order to address these hypotheses, we will take advantage of the Staphylococcus aureus Network Adaptive Platform (SNAP) trial, a multicenter, pragmatic, multi-arm, open-label adaptive platform trial addressing multiple therapeutic questions in patients with S. aureus bacteremia. We will focus in the MSSA “domain” that evaluates the effectiveness and safety of cefazolin vs ISP in a randomized fashion, currently enrolling in Australia, Singapore, Canada, Israel, New Zealand, United Kingdom, United States, Colombia and Chile. The specific aims of our proposal are: i) to define the clinical impact of the CzIE in MSSA bacteremia and, ii) to determine the clinical value and feasibility of a rapid test to detect the CzIE. Our findings are likely to transform the treatment approach for MSSA infections and will provide the basis to develop novel diagnostic tools to the management of MSSA infections.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY Carbapenem resistant Klebsiella pneumoniae (CR-Kpn) have been identified by the World Health Organization as a critical threat on the Priority Pathogens List. Novel diagnostics and therapeutic drug discovery have largely been focused on carbapenemase-producing isolates, however, non-carbapenemase producing carbapenem resistant K. pneumoniae (NCP-Kpn) account for a growing proportion of CR-Kpn clinical isolates. Clinical data on the outcomes of infections due to NCP-Kpn is limited, however, recent studies suggest that rates of mortality may be equal to or even higher than infections due to carbapenemase producing isolates. Carbapenemase independent carbapenem resistance in Kpn has been associated with the alteration or loss of outer membrane porins OmpK35 and OmpK36, and increases in the expression or gene copy number of Extended Spectrum β-lactamases (ESBLs) or plasmid encoded cephalosporinases (AmpC). These changes compromise the entry and concentration of carbapenems in the periplasmic space, and disproportionately impact ertapenem as compared to meropenem or imipenem. In an international cohort of NCP-Kpn isolates, susceptibility to individual carbapenems varied substantially, with all isolates resistant to ertapenem, 67% resistant to meropenem, and 25% resistant to imipenem, respectively. The majority of these isolates had alteration or disruption of the OmpK35/OmpK36 porins with increases in β-lactamase gene copy number. However, the specific drivers that accounted for the discordance between meropenem and imipenem remain to be elucidated. Further, increases in gene copy number expansion also correlated with increasing minimum inhibitory concentration of the β-lactam/β-lactamase inhibitor (BL/BLI) ceftazidime-avibactam, suggesting newer BL/BLI combinations may also be vulnerable to these mechanisms. This proposal seeks to investigate the hypothesis that the specific combination of porin mutations along with the presence, Ambler type, and gene copy number of β-lactamases mediates the differential in vitro phenotypes and can inform optimal treatment selection for NCP-Kpn. In the first aim, a large international collection of NCP-Kpn isolates will be characterized using whole genome sequencing, to investigate the genomic factors associated with discordant carbapenem susceptibility phenotypes. Using a 1-compartment in vitro model, isolates from each NCP-Kpn class will be evaluated for the emergence of resistance to carbapenems and BL/BLI combinations. In the second aim, the clinical impact of NCP-Kpn will be assessed in the cohort of patients with bacteremia. Using a desirability of outcomes ranking (DOOR), differences in mortality, relapse, and emergence of resistance will be compared across infections due to isolates stratified by meropenem susceptibility and underlying mechanism of resistance. The data from this proposal will help to inform the rational selection of antibiotics for treatment of NCP-Kpn infections, and form the basis for the design of larger prospective trials in the future.

NIH Research Projects · FY 2025 · 2025-03

Project Summary/Abstract This R13 resulted from discussions between investigators in the fields of neurostimulation and neuroplasticity at the Houston Methodist Research Institute (HMRI). In 2017, we held an R13-funded pilot workshop that combined research in neural stimulation with new insight into the molecular understanding of neural plasticity and regeneration. The workshop received a strong, positive response by participants and interest in further catalysts for working collaboration. Thus, in 2019, we fostered relationship development and crossover opportunities among attendees via a blitz session wherein newly formed (i.e., established at the 2019 Workshop) collaborative teams competed for two $5K mini grants that supported travel to and from each other’s institutions. In 2021, we went 100% virtual due to COVID-19, however, this did not diminish the R13-funded Workshop’s impact. In 2023, our R13-funded workshop returned to an in-person event, bringing with it many of the successful virtual tools from 2021. On March 4th – 6th, 2025, we will host the 5th Patricia Levy Zusman International Workshop on Neuroregeneration (Zusman Workshop), which will focus on the intersection of electrical activity, brain connectomics, and molecular neural plasticity. Our specific aims are to: 1) formulate new ideas to fill the gap between physiology and functional-based brain stimulation technologies and the molecular and cellular understanding of innate neuronal plasticity; 2) provide promising trainees with various avenues to present their work; and 3) ensure trainees participate in substantial discussions and interactions with faculty members. The 2025 Workshop will be held at HMRI, which is part of the Texas Medical Center (TMC; a one-of-a-kind medical and research hub that fosters cross-institutional collaboration, creativity, and innovation) and is in Houston, Texas (one of the United States’ most diverse cities). A key aspect of the Zusman Workshop is the active inclusion and participation of trainees, particularly women and under-represented minorities. We will encourage their participation via travel scholarships and active promotion and recruitment throughout the TMC. The Workshop promotes the voices of graduate and postdoctoral trainees by including a trainee poster session as well as a session dedicated to trainee oral presentations (given by travel awardees). Further, goal-oriented breakout sessions, led by senior/early-stage investigators and selected trainees, will foster discussion and promote cross-training and collaboration among participants. Overall, this workshop distinguishes itself from established physiology conferences and dedicated neural regeneration conferences by being 1) highly focused on the gap between molecular regeneration and electrophysiology/stimulation, 2) concept driven by clinicians and experimentalists currently problem solving in human therapy, and 3) focused on the establishment of cross training and expertise development in graduate, post-graduate, and clinical fellows. The 2025 Zusman Workshop will ignite nascent collaborations and spur novel multidisciplinary teams to develop innovative, transdisciplinary methods, technologies, and treatments to modulate performance in the damaged nervous system.

NIH Research Projects · FY 2026 · 2025-02

Project Summary/Abstract Nearly one million Americans suffer from a stroke every year, often resulting in serious long-term disability or death. Early detection and treatment of stroke is critical to improving patient outcomes. Several tools have previously been developed that attempt to screen and help identify patients having a stroke, but there are still many stroke cases that are missed, even after the patient arrives to the emergency room. This study aims to improve the identification and triage of stroke patients in emergency departments using artificial intelligence (AI) on multimedia patient data. Our hypothesis is that real-time, standardized, and reproducible stroke assessment tools using AI can improve stroke triage and decrease missed diagnoses in patients with minor-to-moderate neurological symptoms. The study plans to develop a DeepStroke+ augmented-intelligence framework to triage any kind of strokes (ischemic stroke, hemorrhagic stroke, transient ischemic attack, etc) versus stroke mimics commonly seen in emergency settings. We developed a DeepStroke+ framework for stroke triage using facial videos of English-speaking patients who are describing the “Cookie Theft” picture from the Boston Diagnostic Aphasia Examination. Based on this technology, we will develop a stroke triage app and validate it in different triage scenarios from mobile stroke units to emergency room triage in local and telestroke scenarios.

NIH Research Projects · FY 2026 · 2025-01

Project Summary/Abstract: The long-term objectives of this proposal are to reveal new and novel druggable targets and/or elements of fibrogenic pathways, thereby yielding new insights into better treatment strategies for idiopathic pulmonary fibrosis (IPF). IPF is one of the most frequent interstitial lung diseases and has a poor prognosis with worse outcomes than many malignant cancers. The two FDA-approved therapeutic agents (nintedanib and pirfenidone) have entered clinical use recently. Of most concern, while these drugs may slow the decline in lung function to a certain extent, both drugs fail to effectively halt lung fibrosis and improve patient survival. Therefore, there is an urgent need for new and improved therapeutics. In order to achieve higher clinical efficacy, current therapeutic approaches seek drugs that have dual activities against inflammation and fibrosis. To achieve dual activities effectively, we identified a novel central regulator, cyclin- dependent kinase 9 (Cdk9), that controls a common checkpoint for transcriptional activation of genes involved in major inflammatory and fibrotic signaling pathways. Preliminary data show that Cdk9 is elevated in IPF fibroblasts, which drives the aggressive nature of these cells. Treatment with Cdk9 inhibitor reduced the aggressiveness of the cells, and in mouse models of IPF, Cdk9 inhibitor increased survival and reduced fibrosis. These data support the hypothesis that Cdk9 inhibitors can be effective therapeutics to halt or reverse the progression of pulmonary fibrosis. This proposal aims to investigate the mechanistic roles of Cdk9 in IPF and validate Cdk9 as a new anti-inflammatory and anti-fibrotic pharmacotherapeutic target for treating IPF. The approach is to firmly establish a central role of Cdk9 in governing both inflammatory and fibrogenic processes. Our scientific premise rests upon the principle that a potent therapeutic approach to counteract inflammation and fibrosis concurrently can be attained by targeting Cdk9. Three specific aims are proposed, SA1) use gain-and-loss of function analyses to elucidate the molecular mechanism by which Cdk9 regulates IPF pathogenesis and progression, and screen a panel of clinical stage Cdk9 inhibitors for their effectiveness against fibrosis; SA2) test the therapeutic potential of the top performing Cdk9 inhibitor in a bleomycin-induced lung injury mouse model; and SA3) establish therapeutic proof-of-concept by developing an inhalable formulation of Cdk9 inhibitor and demonstrating efficacy in the bleomycin mouse model. Our vision is to generate pre-clinical mechanism-of-action data to support future development of a Cdk9 inhibitor-based IPF therapeutics.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY/ ABSTRACT The increasing incidence of thyroid cancer worldwide is partly fueled by enhanced detection of smaller nodules through imaging techniques like ultrasound, computed tomography (CT), and magnetic resonance imaging (MRI), leading to overdiagnosis of low-risk cancers and benign thyroid conditions. Papillary thyroid carcinoma, the most prevalent type of thyroid cancer, presents lymph node metastasis (LNM) in 60 to 70% of cases, increasing the risk of local recurrence, distant metastasis, and mortality. Fine needle aspiration (FNA), the primary diagnostic method, yields indeterminate results for 10 to 20% of nodules, of which 20 to 30% of nodules may be malignant. Such uncertainty results in repeat biopsies, costly molecular tests, or unnecessary surgeries, causing increased healthcare costs, reduced quality of life, and emotional distress. Core needle biopsies (CNB) are suggested for indeterminate FNA findings. However, CNB is also prone to sampling errors, not suitable near critical structures, and both FNA and CNB are time-consuming procedures and can alter tissue morphology. Given the prevalence of false positives, complications arising from indeterminate diagnoses, prolonged analysis periods, and sampling challenges in thyroid cancer detection, there is a pressing demand for a more effective diagnostic solution. Our proposal introduces an AI-augmented, multimodal, label-free nonlinear optical microscopy system that incorporates coherent anti-Stokes Raman scattering microscopy (CARS), second harmonic generation microscopy (SHG), and two-photon autofluorescence microscopy (TPAF). This innovative system aims for rapid and precise diagnosis of thyroid cancer and LNM. By eliminating the need for external dyes, the label-free technique streamlines the diagnostic process and minimizes complications. This combination of imaging modalities with AI augmentation offers detailed and complimentary subcellular morphological and biochemical information, enhancing our understanding of biochemical composition of thyroid nodules and lymph nodes, facilitating identification of malignancy markers more accurately, and significantly improving diagnostic accuracy. In addition, we plan to develop a compact, portable, AI integrated label-free microendoscope for clinical translation, designed to fit within a core biopsy needle, for differentiating between cancerous and normal tissues and detecting LNM without tissue excision. Validated through studies on ex vivo human tissues and in vivo porcine models, this device promises to transform thyroid cancer diagnosis by serving as a virtual histopathology tool, enabling real-time, label-free imaging and addressing the drawbacks of FNA and CNB, such as overdiagnoses, indeterminate diagnoses, and false positives, without tissue excision or harm to critical adjacent structures. Upon project completion, we anticipate delivering an AI-enhanced, label-free benchtop system for testing in outpatient clinics, capable of imaging FNA samples for rapid and precise diagnosis within minutes, and a comparable label-free microendoscope as a potential optical histopathology device to facilitate near real-time cancer diagnosis without tissue excision.

NIH Research Projects · FY 2026 · 2024-11

Group B streptococcus (GBS) is a major human pathogen that infects diverse anatomic sites and cause wide spectrum of disease manifestations. GBS infections are the leading cause of stillbirths, preterm births and neonatal mortality worldwide. The existing control measures to treat or prevent human GBS infections are significantly challenged by the lack of a human vaccine as well as the rise in antibiotic resistance among GBS strains. Thus, novel translational approaches are urgently required to treat GBS infections. We discovered that a human probiotic produces a previously unknown antibiotic, salivabactin, that is potent in inhibiting the growth of pathogenic streptococci in vitro and in vivo. We also found that the probiotic bacteria produce salivabactin only transiently, which contributes to its reduced probiotic efficacy. To overcome this, we engineered the probiotic to augment salivabactin production and showed its improved prophylactic efficacy in vivo. However, the potency of salivabactin and engineered probiotic in preventing or treating GBS infections remain unknown. The primary objective of this proposal to evaluate the translational potential of two novel strategies, salivabactin and engineered probiotic, to treat or prevent GBS infections in preclinical mouse models of infection simulating various human disease manifestations. We will test our central hypothesis that salivabactin and salivabactin- hyperproducing engineered probiotic are efficacious in controlling GBS infections in two specific aims. The completion of this study will fully assess the translational potential of two novel therapeutic and prophylactic strategies to treat streptococcal infections and identify new antibiotic- and probiotic-based strategies to combat GBS infections.

NIH Research Projects · FY 2024 · 2024-09

Summary/Abstract: Alzheimer's disease (AD) and frontotemporal disorder (FTD) are two of the most common neurodegenerative disorders, with limited available therapeutic options. Although they differ in terms of genes associations, underlying proteinopathy, and affected brain regions, they share pathological features such as increased inflammation and neuronal loss. Recent findings indicate that addressing inflammation might be a potential treatment for AD and FTD. Despite considerable evidence supporting the presence of peripheral and central chronic low-grade inflammation in AD and FTD, a comprehensive molecular understanding of inflammatory cascades and networks across these disorders is lacking. This knowledge gap represents a significant barrier to advancing the field. Our preliminary analysis of inflammation-related analytes from AD, FTD, and healthy control sera, identified the activation of specific inflammatory analytes and signaling nodes in both AD and FTD. In addition to the shared inflammatory signaling nodes, several disease-specific inflammatory analytes were altered in each neurodegenerative disorder. Based on our preliminary results, we hypothesize that the immune responses and activated inflammatory pathways in AD and FTD have shared and disease-specific characteristics and that it may be possible to identify novel interventional strategies for pan-neurodegeneration and disease-specific inflammatory responses. To confirm the preliminary findings and explore novel groups, we will perform a large-scale proteomic analysis to interrogate matched serum and CSF samples from 250 AD, 100 FTD patients and 150 age- and gender- matched healthy controls from two well-characterized national cohorts (Alzheimer's Disease Neuroimaging Initiative (ADNI) and ALL-FTD) (Aim 1). In the next step, we will evaluate the contribution of the altered inflammatory signals to AD and FTD changes. In this regard, we will investigate the association of inflammatory analytes and signaling nodes with 1) known genetic risk factors and mutations, 2) their sex-specific differences, 3) the correlation with relevant established biofluid biomarkers in AD and FTD, and 4) their impact on clinical outcomes (Aim-2). Finally, we will identify the neural and immune cell types responsible for the elevated inflammation in AD and FTD through single-cell RNA-sequencing and spatial transcriptomics (blood and brain tissue) analyses (Aim 3). This multi-omics integrative approach will offer novel insights into the mechanisms driving the peripheral and central inflammatory signals in AD and FTD and suggest treatment options. The innovation in this project results from the collaboration of multiple investigators with experience in neurodegeneration molecular immunology (Alireza Faridar, MD.), neuropathology, biomarkers, and data analysis (Jon B. Toledo, MD., PhD.), and single-cell analysis (Kyuson Yun, PhD.).

NIH Research Projects · FY 2024 · 2024-09

Project Summary: Pulmonary hypertension (PH) is a cardiovascular disorder characterized by high mortality, primarily due to right ventricular (RV) failure (RVF) caused by increased pulmonary vascular resistance. RV fibrosis, a hallmark of decompensated RVF, lacks targeted therapies, highlighting the need to elucidate the molecular mechanisms underlying RV fibrosis and dysfunction. Our research focuses on the role of alternative polyadenylation (APA), a process associated with excessive production of extracellular matrix (ECM) proteins, in RV fibrosis. APA shortens the 3' untranslated region (UTR) of transcripts, leading to loss of microRNA binding sites and increased transcript stability. We have identified Cleavage and Polyadenylation Specific Factor 6 (CPSF6), a key regulator of APA, as being involved in end- stage RVF. In RVF patients, CPSF6 exhibits lengthened 3' UTR and decreased protein expression. Silencing CPSF6 in cardiac fibroblasts (CFs) results in 3' UTR shortening and upregulation of major fibrotic mediators, including TGF-β1 and its receptor, TGFβR1. Pathway analysis further supports 3' UTR shortening in mRNAs encoding ECM proteins in CPSF6 knockdown CFs. Additionally, we have discovered the role of 4-hydroxy-2- nonenal (4HNE), a reactive aldehyde generated during oxidative stress, in RVF. Increased 4HNE downregulates CPSF6, inducing 3' UTR shortening in profibrotic genes and promoting RV fibrosis. The proposed research aims to investigate these mechanisms and identify therapeutic targets for mitigating RV fibrosis. Our hypothesis posits that CPSF6 depletion shortens the 3' UTRs of ECM genes, causing their escape from regulation, promoting their expression, and leading to RV fibrosis. Specifically, we will: Investigate the impact of CPSF6 loss on the 3' UTR landscape and profibrotic gene expression in RVF (Specific Aim 1). Uncover the mechanism underlying CPSF6 reduction-dependent 3' UTR shortening in CFs and its functional consequences in RVF (Specific Aim 2). Assess the impact of ALDH2 restoration on alleviating RV fibrosis through CPSF6 regulation (Specific Aim 3). The validation of our hypotheses and the completion of these aims will highlight the importance of 3' UTR shortening in ECM deposition and fibrosis in RVF, potentially guiding the development of therapeutic interventions. Given the limited treatment options and severe consequences of PH, our research holds significant promise for improving public health.

NIH Research Projects · FY 2025 · 2024-09

Project Summary: Mitral valve prolapse (MVP) is a common valvulopathy affecting 2-3% of the population and nearly 200 million individuals globally. While it is often benign, a subset of patients with MVP may develop arrhythmias, including sudden cardiac death (SCD). Although the association between MVP and SCD was first reported decades ago, the risk was initially believed to be small; contemporary observational studies suggest that SCD may be a more common sequela in MVP, with an estimated yearly incidence between 0.4 and 1.9%. The National Heart, Lung, and Blood Institute recently convened a workshop composed of subject matter experts and stakeholders to identify research needs and opportunities to develop recommendations for the identification and treatment of individuals with mitral valve prolapse, including such individuals who may be at risk for sudden cardiac arrest or sudden cardiac death. Our current proposal is designed to align closely with the priorities identified in the recent NIHBI workshop. Specifically, our proposal has three aims: 1) perform deep phenotyping of patients with MVP to understand characteristics and mechanisms associated with ventricular arrhythmias; 2) Study a blood biomarkers panel (including FDA-approved and novel high throughput proteomic markers) that could be used as a sensitive, costeffective screening strategy to identify MVP patients with myocardial fibrosis who are at risk for SCD; 3) Build a novel SCD risk prediction model in MVP inclusive of CMR evidence of myocardial fibrosis by assembling a large observational multicenter MVP registry with over 2,000 contrast-enhanced CMRs and longitudinal follow-up for arrhythmic events.

NIH Research Projects · FY 2025 · 2024-09

Abstract We discovered that nuclear reprogramming of somatic cells to a different somatic cell lineage, or induced pluripotent stem cells, requires activation of inflammatory signaling within the cell. Specifically, pattern recognition receptors (PRRs) such as toll-like receptors (TLRs) mediate a cell-autonomous innate immune response via NFKb and IRF3. We found that this inflammatory signaling causes global changes in the expression and/or activity of epigenetic modifiers to increase DNA accessibility and fluidity of cell phenotype. Subsequent work has suggested that this process of “transflammation” may be involved in vascular regeneration. Specifically, we have shown that fibroblasts in an ischemic region can be transformed into endothelial cells (ECs) through an “angiogenic transdifferentiation” process. This process contributes to the recovery of perfusion in the ischemic region, as the recovery of the microvasculature, and the restoration of blood flow in an ischemic region is antagonized by factors required for angiogenic transdifferentiation (e.g., inflammatory signaling). My recent work indicates that cell metabolism may be an important contributor to this process. Specifically, a glycolytic shift is induced by inflammatory signaling which is required for the transdifferentiation of fibroblasts to ECs. Thus, regulating cell metabolism within fibroblasts to facilitate their transdifferentiation into reparative ECs may be a novel strategy for treating ischemia. To determine the molecular metabolic pathway that leads to transdifferentiation, we will alter the function of key metabolic enzymes in mice pharmacologically and genetically in vivo to confirm our proposed pathway and demonstrate the metabolic regulation of transdifferentiation in a mouse model of peripheral artery disease (PAD). We will pursue experiments to trace key metabolites and demonstrate their importance in mediating DNA accessibility and transdifferentiation to identify their role in epigenetic regulation in cell fate transition. Completion of these studies will demonstrate the novel concept that metabolic regulation within cells contributes to their fate and provide novel targets to enhance this process for the treatment of PAD.

NIH Research Projects · FY 2024 · 2024-09

Glioblastoma (GBM) is uniformly fatal with an incidence rate of 2.99-3.23 per 100,000 people. Despite recent therapeutic advances, our understanding of tumor biology in GBM remains incurable with median survivals of less than two years. Critical unanswered questions contributing to this dismal prognosis that elude clarification through traditional analysis of human clinical material or small animal models include i) how does surgical trauma impact GBM development and adjacent brain tumor microenvironments, ii) what are the unique cellular and molecular properties of residual infiltrating tumor cells during disease progression, and iii) how do surgically induced selection pressures generate de novo molecular and cellular heterogeneity not present in unresected surgical samples. Development of a cure or at least a treatment that would provide significant quality of life improvements in GBM patients is ultimately limited by the lack of animal models that reproduce the hallmark features of GBM tumor. Typically, rodent models are used to study GBM; however, rodents differ vastly from humans (e.g., brain size and complexity), making surgical interventions difficult to simulate. By contrast, the mini- pig brain is remarkably like the human brain and its larger size permits relevant surgical and imaging studies. Therefore, we propose to develop a first-in-kind syngeneic (intact immune system) glioma model in mini-pigs to recapitulate hallmark features of human glioma. Development of this model will support studies heretofore impossible in rodent models or human patients. Thus far, we have established oncogene activated transformed pig glioma cell lines from pig brains and confirmed their tumorigenic capabilities in mouse models and onco- minipigs. In the proposed research, we will determine the optimum engraftment conditions of these oncogene activated transformed pig cells by implanting them into host pig brain under chronic vs transient immunosuppressive microenvironment. We will also study the impact of provincial treatment of anti-inflammatory and immune suppressive drugs. To eliminate any potential impact of immune activation between different donors and hosts, we will repeat the same approach to induce in vivo viral oncogenesis. The relevance of the proposed syngeneic pig model will be established through analysis of accepted hallmark MR imaging, stereotactic guided intracranial surgery, and pathology features of the human disease, followed by immune phenotyping. Development of a large animal glioma model is expected to facilitate new insight into human glioma biology. As a first step, we propose to test the hypothesis that mini-pig syngeneic glioma models will recapitulate key features of the human disease. Further, these models are expected to provide a robust new platform for future studies not possible in rodent models or practically achievable through analysis of patient-derived material. This includes, but is not limited to, the evaluation of new therapies, imaging studies, and surgical techniques. Finally, development of this large animal model will enable not only us, but the larger scientific community, to answer clinically relevant questions applicable to GBM and other brain metastatic cancers.

NIH Research Projects · FY 2024 · 2024-08

Abstract Neurogenic lower urinary tract dysfunction (NLUTD), formerly “neurogenic bladder,” encompasses bladder and sphincter abnormalities resulting from neuromuscular diseases or injuries. Conditions such as cerebrovascular accidents (CVAs), spinal cord injuries (SCIs), and age-related neurogenerative diseases like Alzheimer’s and other types of dementia contribute to NLUTD1. This condition is prevalent, affecting approximately 15% of stroke patients and 70-84% of those with spinal cord injuries2. Furthermore, NLUTD is found in 40-90% of individuals with Multiple Sclerosis (MS), 37-72% in those with Parkinson’s Disease (PD) and has been reported in more than 90% of children with spina bifida3,4. Urinary incontinence in individuals with dementia escalates in tandem with the severity of the condition, and except for SCI, aging is the primary risk factor for these neurological conditions5-7. Patients with NLUTD experience drastic effects on quality of life secondary to the development of urinary tract infections (UTIs), urinary retention, incontinence, renal dysfunction, and autonomic dysreflexia1. The Neurogenic Bladder Research Group (NBRG) is an academic society dedicated to improving the lives of patients with NLUTD through application of patient-centered outcomes research. Sequelae of NLUTD are usually electively managed with a combination of medications, medical devices and surgical approaches. Recently, more advanced devices have been employed for management of urinary symptoms associated with NLUTD, including tibial nerve stimulators and sacral neuromodulation systems. However, while these devices improve the quality of life for patients with neurogenic urinary symptoms, further research and development is critical to maximize the efficacy and longevity of urologic technology8,9. The overarching goal of this proposal is to host an NBRG affiliated meeting, which will be the first of its kind, to identify areas in the management of NLUTD that would benefit from engineering solutions. To achieve this theme, in Aim 1, leading researchers in engineering and multiple medical specialties, including urology, neurology, neurosurgery, and pediatrics – as well as patient representatives – will be invited to participate in active lectures, group discussion, round tables to identify the gaps in NLUTD care that engineering solutions can address. In Aim 2, guidelines will be written to outline the role of engineers in neurogenic bladder care and guide the development of engineering solutions in NLUTD for the near future. In Aim 3, engineering, doctoral, and medical students and junior investigators will be invited to submit abstracts for presentation to promote early showcasing and mentorship in NLUTD research. Toward the manifestation of these aims, we are applying for an R13 conference grant. This timely meeting is in full agreement with the missions of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) and the National Institute on Aging (NIA). This conference will the first to bring together engineers and researchers/clinicians to advance technologic solutions for the care of NLUTD and to compose a series of guiding principles on the subject.