Methodist Hospital Research Institute

NIH Research Projects · FY 2025 · 2023-09

The overall goal of this project is to develop novel mathematic methods and toolkits to connect cell fate transition and epigenetic regulation across tissues and diseases. Cell fate transition often occurs in organ development, tissue regeneration, and pathogenesis. Dysregulation of the cell fate transition can lead to abnormal development or diseases, such as type 2 diabetes, obesity, heart failure, and Alzheimer’s disease. Quantitively decoding how cell fate changes can provide novel mechanistic insight into organogenesis and tissue regeneration, and help identify new strategies for the treatment of human diseases. However, our knowledge of cell fate transition and its regulation is only the tip of the iceberg due to the impracticality of long- term tracing of cell transcriptomes. In the past decade, numerous single-cell atlases containing millions of cells in different tissues, organs, developmental stages, and biological conditions are routinely developed by consortia such as the Human Cell Atlas (HCA). These atlases provide an opportunity for the unbiased study of cellular dynamics and the regulation mechanism. The lack of computational methods presents a major knowledge gap in the understanding of cell dynamics and the regulation leveraging by those large reference atlases. To address this knowledge gap, we proposed a new concept of “reference-based cellular dynamic inference”, which is a novel strategy to automatically annotate the cell state transition in new datasets by learning from the appropriate reference, allowing us to easily perform comparative analysis among different tissues and disease conditions. In this project, we will pursue three parallel but complementary research directions: 1) to develop the first computational methods and toolkits for generating cell dynamics atlases and analyzing cell state transition based on the appropriate reference atlases; 2) to develop novel statistical models for studying epigenetic regulation of cell fate from single-cell multiomics data; 3) to generate the first dynamic reference landscapes of cell differentiation, such as cardiogenesis, hematopoiesis, and neurogenesis, and in- house landscapes of transdifferentiation. This project will be built on the foundation of our recent studies for the development of computational approaches to uncover cell state transition from single-cell transcriptomes in both homogeneous and heterogeneous cell populations and the studies for investigating the role of epigenetic regulation on cell fate transition. The proposed studies will generate advanced computational toolkits and broadly applicable dynamic reference atlases, which are expected to reveal profound mechanisms controlling cell state transition in health and disease. In the long term, the ability to build cell dynamics reference landscapes will open a new horizon to understand the diversity of cell fate through comparative analyses across tissues and diseases and enhance regenerative medicine.

NIH Research Projects · FY 2026 · 2023-08

PROJECT SUMMARY The extracellular matrix (ECM) provides a scaffold for cardiac structural integrity and plays an active role in modulating cellular responses. ECM consists of a network of fibrillar proteins and proteoglycans binding cyto- kines, growth factors, and ECM-modifying enzymes. The aging heart develops interstitial fibrosis, and prelimi- nary data demonstrate that different proteins are enriched in ECM in the male and female heart. An altered ECM can affect cellular phenotypes. The ECM-cell communication occurs via cell surface receptors such as integrins, through outside-in signaling, where integrin-binding to ECM protein domains translates into prolifera- tion, differentiation, or gene expression. On the other hand, activation of the intracellular integrin tail increases their ECM-binding affinity, promoting adhesion, migration, or ECM assembly (by inside-out signaling). There- fore, ECM dictates a cellular phenotype, but cells can modify ECM in response by changing its composition or assembly. Because of profound differences in ECM composition in males and females, we hypothesize that the quality of repair assembled after injury and subsequent adverse remodeling in males and females will also be differ- ent, and the differences will be accentuated by aging. In SA1, we will map the differences in ECM dependent on age (young and old animals will be studied), sex hormone level (animals will be subjected to chemical ovary failure or castration) or level of inflammation (CCL2KO animals will be used; these mice have reduced leuko- cyte infiltration and therefore reduced fibrosis). The cellular phenotype of fibroblasts, endothelial cells (ECs), and smooth muscle cells (SMCs) isolated from all these animals will be studied ex vivo and in 3D culture. The effect of ECM on cell phenotype will be tested in 3D cultures using the matrix swap approach, where i.e. young cells will be cultured on ECM from the old hearts and vice versa. And the effect of matrix stiffness on the pro- duction of ECM protein will be examined as well. Similarly, the physiology of human cells will be studied. In SA2, we will examine ECM-integrin dependent phenotypic changes in mouse and human cells. We will first examine the effect of sex hormones, age, and inflammation on the levels of integrin expression in fibroblasts, ECs, and SMCs. Then we will manipulate the integrin expression level (via downregulation or overexpression), and examine the cellular phenotype. Finally, we will manipulate estrogen and androgen receptors and deter- mine the effect of these manipulations on integrin levels and ECM protein synthesis. In SA3, we will examine how sex hormone levels and age affect scar formation and adverse remodeling after ischemia/reperfusion inju- ry. Changes in ECM will be examined via ELISA, mass spectrometry, and western blot. The approach is inno- vative because it examines the role of three superimposed variables (age, sex hormone, and inflammation) on ECM and cellular phenotypes. Successful completion of these studies will provide a mechanistic link for future therapeutics targeting insufficient reparative and excessive adverse fibrosis.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY The American Consortium of Early Liver Transplantation-Prospective Alcohol-associated liver disease Cohort Evaluation (ACCELERATE-PACE) study is a prospective longitudinal cohort of patients with severe alcohol- associated liver disease (ALD) evaluated for early liver transplantation (ELT). The cohort leverages the ACCELERATE consortium with 4-linked R01s and 5 additional recruitment sites in the South/Southeast, Mid- Atlantic, Midwest, and West, and will refine and identify best practices in the selection and management of patients with severe ALD considered for ELT across their continuum of care. ALD is now the most common indication for liver transplantation (LT) in the U.S. Historically, LT centers required at least 6 months of alcohol abstinence before LT referral and evaluation, though empiric evidence to support minimum sobriety periods was limited. ELT, defined as LT before 6 months of abstinence, is increasingly performed but with significant practice variability. There is no consensus on optimal ELT candidate selection, and selection criteria vary widely, contributing to disparities in access to lifesaving care. ELT is also controversial due to the potential for liver recompensation with abstinence, which would obviate the need for LT—accurate prediction of recompensation has the potential to increase organ utility and stewardship. Detailed evaluation of the efficacy of alcohol use disorder treatments and improved risk scores based on pre-LT psychosocial factors to predict return to alcohol use are needed to refine selection criteria, optimize post-LT care, and effectively treat AUD. Short- and intermediate-term survival after ELT is excellent, but the incidence and predictors of post-LT complications are poorly defined. To fill these key knowledge gaps, we will enroll and prospectively follow 770 ELT candidates and 270 ELT recipients for 3 years at 9 socio-demographically diverse centers. The proposed Aims will: (i) inform ELT selection criteria and investigate potential sources of bias in ELT evaluation and healthcare disparities in ELT access; (ii) develop risk prediction scores for LT-free survival and recompensation; (iii) identify effective treatments (medical, behavioral) for alcohol use disorder among patients with severe ALD and post-ELT; (iv) evaluate clinical outcomes among ELT candidates and recipients, including mortality, transplantation, post-LT complications (e.g. cancer, cardiovascular events, graft rejection/failure), and quality of life. A comprehensive data repository will include sociodemographic, clinical, geospatial, psychosocial, behavioral, and patient-reported outcome variables. LT documents, checklists, recordings of selection meetings, direct observations of LT procedures, and clinician interviews will identify best practices and pitfalls in candidate selection. A biorepository of blood, urine, explant/donor tissue, pre- and post-LT liver tissue, peripheral blood mononuclear cells, and cross-sectional radiologic imaging will inform future ancillary studies.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY Blood CD8+ T cells can be subdivided into different subsets based on their cytokine section profiles and functions. We have shown that IL-9-secreting CD8+ Tc9 cells mediate a stronger antitumor effect in vivo compared to the classical IFN--secreting Tc1 or CTLs. However, the underlying mechanisms remain unclear. Recently, we discovered that Tc9 cells could eradicate large-established tumors by inducing an enhanced tumor pyroptosis, a form of programmed cell death dependent on caspase-1 activation. Our preliminary studies showed that Tc9 cell-treated tumors had increased expressions of IL-1 and IL-18 and activated caspase-1, and more tumor cell death compared to Tc1-treated tumors. Neutralizing IL-1 and IL-18 in vivo completely abrogated the therapeutic advantage of Tc9 cells over Tc1 cells in tumor controls. We further showed that tumor-specific Tc9 cells killed tumor cells by inducing both caspase-1-depedent pyroptosis and caspase-3- dependent apoptosis while Tc1 cells mainly induced apoptosis in tumor cells. Similarly, human tumor-specific Tc9 cells also displayed stronger antitumor effects in vivo compared to Tc1 cells, which was IL-1- and IL-18- dependent. Interestingly, we observed that IL-1 plus IL-18 induced apoptosis in cultured Tc1 but not Tc9 cells, Tc9 cell-derived IL-9 is critical for Tc9 cell activation of STAT3 and pro-growth and survival signaling and function, and IL-1 plus IL-18 can re-activate STAT3 and pro-growth and survival signaling in aged Tc9 cells with reduced IL-9 production. Thus, these findings reveal novel mechanisms underlying how Tc9 cells overcome the suppressive tumor microenvironment after transfer, survive and persist longer, and exert a greater antitumor activity as compared to the traditional Tc1 cells. We hypothesize that tumor-specific Tc9 subset may be superb effector T cells for cancer immunotherapy due to their capacity to induce both pyroptosis and apoptosis in tumor cells and utilize Tc9 cell-secreted IL-9 and tumor-produced IL-1 and IL-18 for their fitness, longevity, and function in vivo to effectively eradicate large established tumors. To test our hypothesis, Aim 1 will determine the importance and mechanisms of Tc9 cell-induced tumor cell pyroptosis in tumor clearance via IL-9- and GrzB-induced NFB-NLRP3-caspase-1-gasdermin activation, and Aim 2 will determine the role and mechanisms underlying Tc9 cell persistence and function mediated by Tc9 cell- secreted IL-9 and tumor-produced IL-1 and IL-18. Completing this project will reveal the importance and mechanism in induction of tumor pyroptosis by tumor-specific Tc9 cells and elucidate the mechanisms underlying the longevity and function of Tc9 cells in tumor microenvironment.

NIH Research Projects · FY 2025 · 2023-07

ABSTRACT Metaplastic breast cancer (MpBC) is a rare subset accounting for <5% of all breast cancers. MpBC is a significant health challenge as it exhibits the most dismal prognosis of all breast cancer subtypes, worse than non-MpBC triple-negative breast cancer (TNBC), with median survival rate of 8 months or less in patients with metastatic disease. Due to a lack of druggable targets, the main therapeutic option for metastatic MpBC remains systemic chemotherapy, despite known resistance to most cytotoxic drugs. One common molecular alteration in MpBC is hyperactivation of the phosphoinositide 3-kinase and protein kinase B (PI3K/AKT) pathway. Additionally, we published that MpBC also displays a gain-of-function oncogenic mutation in ribosomal protein L39 (RPL39), which is responsible for treatment resistance, stem cell self-renewal, and lung metastasis. The mechanistic function of RPL39 is mediated through inducible nitric oxide synthase (iNOS)-mediated nitric oxide production. In a recently published clinical trial targeting this nitric oxide synthase (NOS) pathway with a pan-NOS inhibitor NG-methyl-L-arginine acetate (L-NMMA), high efficacy in chemorefractory TNBC patients was demonstrated. Furthermore, in vivo studies performed showed a significant reduction in tumor growth, associated with a significant increase in apoptosis after the alpelisib/L-NMMA combinatorial regimen. Therefore, we hypothesize that the NOS and PI3K signaling pathways may exert their oncogenic responses synergistically to promote aggressive tumor growth. To test this hypothesis, Specific Aim 1 seeks to demonstrate the therapeutic efficacy of simultaneous inhibition of NOS and PI3K pathways with chemotherapy in MpBC pre- clinical models on primary tumor growth and metastasis. Specific Aim 2 will investigate the global and RPL39- specific ribosome translation landscape in response to NOS/PI3K inhibition in MpBC. In Specific Aim 3, the cell- cell interactions among tumor cells, myeloid cells, lymphoid cells, and stromal cells within the tumor microenvironment and their role in supporting cancer niche populations will be evaluated at the single-cell level using spatial transcriptomics, immunofluorescence, CyTOF imaging systems, and a multi-modal data analysis model. This study thus proposes a mechanistic investigation of a combinatorial targeted approach against the two key pathways in MpBC, identifies cell–cell interactions, and develops unique crosstalk models that will effectively predict outcome and treatment response and complement our recently funded U01 clinical trial on MpBC patients.

NIH Research Projects · FY 2025 · 2023-06

Project Summary Abstract: Segmental demyelination is a pathologic process of stripping off the myelin sheath, which shunts current out of axons and results in the failure of action potential propagation in a variety of peripheral nerve diseases. However, the molecular program that leads to demyelination remains unclear, which hampers the therapeutic development for demyelinating neuropathies. Humans with autosomal recessive mutations in the FIG4 gene develop Charcot-Marie-Tooth disease type-4J (CMT4J), a disease characterized by segmental demyelination. Our preliminary studies in a CMT4J mouse model (Fig4−/−) have demonstrated that segmental demyelination is associated with increased intracellular Ca2+ in myelinating Schwann cells and accumulation of macrophages in the spinal roots. Administration of a Ca2+ chelator suppresses the demyelination in Fig4−/− mice. Therefore, this mouse is an appropriate model to investigate the molecular events underlying demyelination. Toward this end, we will test our central hypothesis that signals from FIG4-deficient axons and/or macrophages exacerbate the overload of intracellular Ca2+ in FIG4-deficient Schwann cells which, in turn, leads to segmental demyelination. We propose three Specific Aims to test our hypothesis by determining if: (1) FIG4-deficient Schwann cells are sensitized to demyelinate upon challenge with Ca2+; (2) FIG4-deficient macrophages release cytokine IL12B that triggers a further increase in intracellular Ca2+ in FIG4-deficient Schwann cells, leading to demyelination; and (3) individual proteins in the PAS complex play distinct roles in the development and maintenance of myelin and axons. These studies have the potential to uncover novel molecular mechanisms underlying demyelinating peripheral neuropathies and identify targets for therapeutic development.

NIH Research Projects · FY 2025 · 2023-05

Abstract Radiofrequency (RF) ablation of ventricular tachycardia (VT) in ischemic cardiomyopathy is fraught with limitations due to suboptimal efficacy, risk of complications, and frequent need for repeat procedures. Ischemic VT arises as a result of reentrant circuits within or around the myocardial scar of an infarct. The co-localization of ventricular arteries, veins, and nerves, is an anatomical fact, determined by the embryology of coronary vessels. Just as myocardial infarctions have a “culprit” or “infarct-related artery”, there commonly exists an “infarct-related vein” or veins in VT substrate. Additionally, it is well known that autonomic innervation -in anatomical proximity to the veins- plays an important role in post-MI arrhythmogenesis. We have developed an approach to target ablation-refractory VTs via ethanol delivery in the coronary veins that provide venous return from arrhythmogenic sites (venous ethanol, VE). Beyond an initial set of case reports, we have validated the utility of VE in a large, multinational registry, in which we establish the safety and efficacy of VE in RF-refractory VT. Given the co-localization of epicardial arteries, veins and nerves, VE may be particularly suited to impact infarct innervation. Thus, a central goal of this proposal is to capitalize on the presence of coronary veins on the epicardial aspect of a myocardial scar as a therapeutic opportunity of unique mechanisms. We hypothesize that VE added to conventional catheter ablation improves the results of VT ablation. We propose a single-site, investigator-initiated clinical trial on VE. In Aim 1-R61 phase-, we propose to finalize the design of randomized clinical trial to assess the clinical efficacy, and safety of VE when used in combination with RF ablation compared with RF ablation alone -Venous Ethanol for Left Ventricular Ischemic VEntricular Tachycardia -VELVET clinical trial. The trial will include an investigational new drug (IND) authorization by the FDA. Patients with ischemic VT will be randomized to conventional endocardial ablation alone, vs combined with VE in the infarct-related vein. In Aim 2 -R33 phase- we will enroll a total of 156 patients, and collect efficacy, safety and procedural data on the impact of VE added to catheter ablation. This trial will allow for a wealth of new imaging data to be collected that will characterize the extent of myocardial scar and innervation before and after VE -compared to endocardial RF alone. In Aim 3 -R33 phase- we will collect multi-modality imaging data characterizing the VT substrate before and after ablation -with catheter ablation alone vs combined with VE. Cardiac magnetic resonance, venous CT angiograms and regional adrenergic innervation maps with positron emission tomography (PET) scans of innervation tracers (11C hydroxyephedrine, 11C-HED) will provide a complete structural assessment of the VT substrate, before and after ablation. If completed, the project will validate a new procedural strategy and will provide key new insights into the structural determinants of VT ablation success.

NIH Research Projects · FY 2026 · 2023-04

Atherosclerotic cardiovascular disease (ASCVD) and plasma high-density lipoprotein cholesterol (HDL-C) are negatively correlated. One model assigns this correlation to the role of HDL as an acceptor of free cholesterol (FC) transfer from arterial-wall macrophages (FC efflux), and in some studies, FC efflux better predicted ASCVD than HDL-C concentration. However, interventions that increase plasma HDL failed to reduce ASCVD, and, paradoxically, several studies revealed higher ASCVD mortality among patients with very high HDL. The source of this paradox is unknown, appropriate therapies have not been formulated, and in this context, the role of the reverse process, HDL-FC influx, which may be supported by excess HDL-FC, is unknown. Mice deficient in the HDL-receptor, Scarb1-/- mice, are a robust model of the human high-HDL phenotype. Compared to wild-type mice, Scarb1-/- mice are more athero-susceptible and have higher plasma levels of HDL that is more FC-rich. This produces a state of high HDL-FC bioavailability, which we express as an index: HDL- FCBI = HDL particle number (HDL-P) x mol% HDL-FC. This conceptually new metric was first reported by this study team, which reported that HDL-FCBI increases as wild-type mice < human << Scarb1-/- mice and that more FC transfers from HDL from Scarb1-/- vs. wild-type mice to macrophages. Thus, we hypothesize that similar mechanisms underlie ASCVD among humans with very high plasma HDL-C. Our goal is to compare patients with positive (CACS>0) and negative (CACS=0) coronary artery calcium scores respectively assigned as ASCVD and non-ASCVD in three subgroups—those with high (HH), intermediate (IH), or optimal (OH) plasma HDL-C concentrations—and test whether ASCVD is associated with a high HDL-FCBI. According to our hypothesis, a) HDL-FCBI will be higher among HH vs. OH patients and among ASCVD vs. non-ASCVD patients, especially those with high plasma HDL-C, and b) the magnitude of FC transfer from HDL from ASCVD patients to macrophages will be greater than that from HDL from non-ASCVD patients, again especially among ASCVD patients with high plasma HDL-C. This hypothesis is supported by studies of Scarb1-/- mice in which a component of HDL-FCBI is reduced and with it, ASCVD—reducing HDL-P with probucol or mol% HDL-FC by increased FC esterification suppressed ASCVD. Comparison of CACS vs. HDL- FCBI of all three groups will reveal a non-linear functional relationship. Traditionally, physicians have measured HDL and LDL in terms of their total cholesterol content. These measures have had good but imperfect predictive value, mainly because the two components of TC, FC and cholesteryl esters, have distinct metabolic itineraries that may differentially contribute to ASCVD. Validation of the study-hypotheses in humans would provide a compelling rationale for measuring plasma lipoprotein FC as an ASCVD diagnostic and for the formulation of therapies that reduce plasma- and especially HDL-FC.

NIH Research Projects · FY 2026 · 2023-04

Project Summary Adoptive cell therapy is a promising approach to treat cancer, but despite tremendous efforts, results of clinical trials in human solid tumors using ACT with tumor-specific T cells expressing T-cell-receptors (TCR) or chimeric antigen receptors (CAR) have not demonstrated the desired therapeutic responses. Recently, we developed tumor-specific T cells loaded with the myxoma virus to overcome resistance in solid tumor ACT. We hypothesize that anti-solid tumor activity of these T cells is mainly attributed to a special type of tumor cell death, autosis, that has not been considered a T cell killing mechanism before but may contribute to the observed exciting antitumor potency by targeting both antigen-positive and antigen-negative tumor cells. Aim 1 will determine the unique molecular mechanism underlying tumor cell autosis induced by the synergy of myxoma virus-derived factor(s) with T cell-derived soluble factor(s). Aim 2 will determine the role of these tumor-specific T cells in promoting autosis and robust host immunity to eradicate solid tumors when the targeted antigen is expressed by only ~25% of tumor cells. Our proposed studies will identify a novel ACT strategy endowed with the capacity to eliminate solid tumors with very high antigen heterogeneity. This translationally relevant work holds promise to significantly advance the therapeutic index of ACT in solid tumors and could then lay the foundation for future clinical trials.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Maternal mortality and AIDS are among the leading causes of death in reproductive-age women globally due to unintended pregnancy and the disproportionate burden of HIV infection. While effective prevention methods exist, only 21% of women have access to modern contraceptives and adherence to HIV PrEP regimen is poor in many HIV-endemic regions. Multipurpose prevention technologies (MPT) aim to synchronize contraceptive delivery with HIV PrEP, offering a patient-centered approach to provide dual coverage to sexually active women in a single product. A long-acting unified delivery product that can be administered in a discreet manner could reduce stigma around contraceptives and HIV PrEP, thereby improving uptake and adherence to combination prevention products. Our goal is to develop a long-acting delivery implant of etonogestrel (ENG) and islatravir (ISL) drugamer with an unprecedented 2-year release duration for pregnancy and HIV prevention that eliminates further need of adherence. A new subcutaneous nanofluidic multipurpose implant (NanoMPI) will be brought together with a new polymeric prodrug technology termed “drugamer”, to achieve this innovative product candidate that delivers ultra-long acting ENG+ISL drug dosing durations. The NanoMPI leverages a nanofluidic membrane with nanochannels and a single drugamer reservoir to achieve zero-order release kinetics through passive diffusion without pumping mechanisms, permitting discreet, user-independent dosing. Key product attributes will include drug stabilization on the multi-year timeframe, minimized early- and late-stage burst release and pharmacokinetic (PK) tailing, zero-order drug release kinetics tailored to the individual drug PK/PD (pharmacodynamic) requirements, user-independent dosing, and retrievability. The project is structured around 3 specific aims: 1) to develop optimized ENG-ISL drugamer formulations that stabilize the drugs and exhibit the desired zero order release profiles, in concert with the optimization of the NanoMPI device for 2-year sustained release; 2) to assess pharmacokinetics, tolerability, and safety of the lead ENG-ISL NanoMPI device for 2 years in pigtail macaques; and 3) to evaluate contraception and HIV PrEP efficacy of ENG-ISL NanoMPI in pigtail macaques via repeated low-dose vaginal challenge and develop a PK/PD model. We will use a milestone-driven research approach to advance the ENG-ISL NanoMPI technology towards the ultimate goal of clinical translation, leveraging the interdisciplinary team’s experience in drugamer technology, drug delivery, pre-clinical and clinical HIV prevention studies, and hormonal contraceptive and antiretroviral pharmacology. Importantly, the NanoMPI addresses user preferences for a dual prevention product, discretion and longer dosing duration. Successful completion will generate a versatile MPT platform that could be adopted for other drug combinations and preventative strategies.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Atrial fibrillation (AF) is the most common sustained adult arrhythmia, associated with an increased risk of stroke, heart failure and dementia. With increased longevity in chronic diseases, the prevalence of AF – currently estimated at 46 million worldwide – is dramatically rising. Catheter ablation – tissue destruction – is the most effective therapy but is fraught with procedural risks and suboptimal efficacy. The cardiac autonomic system (CANS) is known to be involved in the pathogenesis of AF, but no specific diagnostic or therapeutic approaches have evolved from this. Our long-term goal is to devise neuromodulatory AF treatment and preventive strategies and fill the knowledge gap on the mechanisms by which CANS neuronal and humoral paracrine output modulate atrial function in humans. We break through existing technical barriers that limited our understanding of intrinsic cardiac ganglia, capitalizing on the vein of Marshall as a vascular route to sample its electrophysiology and humoral responses, collecting atrial coronary circulation blood, and recording nerve activity from the ganglionated plexi (GP). Our extensive preliminary data in patients shows that apnea increases GP activity measured using novel percutaneous technology in AF patients undergoing ablation procedures. Remarkably, we found that Substance P (SP) collected from the coronary sinus is elevated compared to undetectable levels in peripheral blood of AF patients suggesting that GP activation via secreted SP may play a role in AF substrates. In large animal models, we have found that specific ablation of GP sensory neurons blunts the pro-fibrillatory response to apnea and that a crescendo GP response occurs after repeated consecutive apneas. Our data in human pluripotent stem cell-derived atrial cardiomyocytes (hiPSC-aCM) shows that chronic SP treatment affects cardiomyocyte electrophysiology and modifies gene expression of miR-21 targets. These exciting observations, innovative methods, and unique clinical and basic science expertise position our team to develop this project successfully. We propose the central hypothesis that CANS produces a substrate for AF through neural (nerve firing) and humoral effects (secretome), in which SP – released by GP sensory neurons – plays a major role in increasing susceptibility to AF through direct electrophysiological and genomic effects in atrial cardiomyocytes. The central hypothesis will be tested by pursuing studies 1) in humans with paroxysmal and persistent AF aiming to measure nerve activity and secretome of intrinsic ganglia through the vein of Marshall during ablation procedures, 2) in canine models of acute and persistent AF to determine whether ablation of GP or SP antagonism ameliorates AF, 3) in hiPSC-aCM and engineered atrial tissues to elucidate SP actions. The proposed research is significant because it is expected to provide a mechanistic understanding of the relationship between CANS and sleep apnea for the continued development of effective therapies against AF. Ultimately, such knowledge can offer new opportunities to develop innovative therapies to treat AF.

NIH Research Projects · FY 2026 · 2023-03

ABSTRACT: The combinational antiretroviral therapy is effective in suppressing HIV-1 replication. However, latent HIV reservoirs cannot be cleared by such antiretroviral therapy, as viral rebound develops rapidly after the treatment is interrupted in people living with HIV. The shock-and-kill approach to eliminate HIV-1 reservoirs is to induce viral reactivation using latency reversal agents and trigger cell death through virus-mediated cytopathic effects or immune-mediated clearance. However, shock-and-kill by latency reversal agents has not been shown to successfully reduce the viral reservoirs in HIV-1 patients. It has been reported that latency reversal agents can up-regulate pro-survival autophagy and anti-apoptotic molecules that counteract cell death signaling. Inhibition of these pro-survival mechanisms can promote the killing of HIV-1-infected cells, and facilitate the killing of HIV-1 reservoir cells during vial re-activation. Experiments are proposed to test hypothesis that cellular mechanisms that maintain the long-term persistence of HIV-1 reservoirs are important for protecting the reservoir cells against cell death, and targeting these cellular mechanisms can sensitize HIV-1 reservoirs to cell death during viral reactivation: 1) To determine the mechanisms for the specific induction of cell death in HIV-1 reservoirs in response to viral reactivation. The mechanisms for the induction of apoptosis and alternative cell death pathways in latent HIV-infected T cells by latency reversal agents will be tested; 2) To delineate the mechanisms that confer the resistance to cell death in HIV-1 reservoirs upon viral reactivation; and 3) To test whether targeting pro-survival mechanisms sensitizes HIV-1 reservoirs cells to cell death during viral re-activation. The studies will reveal novel molecular pathways that could be targeted to sensitize HIV-1 reservoirs to cell death, and facilitate the development of an effective approach to clear HIV-1 reservoirs.

NIH Research Projects · FY 2026 · 2023-02

Project Summary/Abstract Chronic lymphocytic leukemia (CLL) represents 30% of adult leukemia and is still incurable. According to the National Cancer Institute, approximately 21,250 new cases of CLL and 4,320 deaths from CLL are projected in the United States alone in 2021. Although the Bruton's tyrosine kinase inhibitor (BTKi) is an effective targeted therapy for CLL, approximately 50% of BTKi-treated CLL patients have dropped out of the therapy due to chronic adverse effects. Additionally, significantly increased numbers of CLL patients treated with BTKi or other kinase inhibitors have developed the more medically challenging diffuse large B cell lymphoma. Novel therapy that can directly target CLL or be rationally combined with BTKi is highly desirable. The stimulator of interferon genes (STING) is an endoplasmic reticulum (ER)-resident protein critical for sensing cytoplasmic DNA and promoting production of type I interferons, thereby boosting immune responses. STING agonists, such as ADU-S100, have been used as combination immunotherapy with Pembrolizumab in clinical trials to treat advanced solid tumors and lymphomas, and these therapeutic applications of STING agonists are based on the main known function of STING in eliciting anti-tumor immunity. We were the first to show that STING agonists directly induce potent mitochondria-mediated apoptosis in B cell-derived malignancies including CLL, while these agonists induce production of interferons in melanoma, hepatoma and lung cancer cells without suppressing their growth. Apoptosis of CLL, B cell lymphoma and multiple myeloma requires STING, because genetic deletion of STING results in resistance of all these cells to STING agonist-mediated apoptosis. Apoptosis of these cells is not due to the production of inflammatory cytokines but may involve the prolonged presence of agonist-bound STING. Together with our new preliminary results showing that mutation-mediated activation of STING causes rapid degradation of the B cell receptor (BCR) and significantly reduced BCR signaling (disadvantageous for CLL survival), and that novel serine residues on STING are phosphorylated in agonist-stimulated malignant B cells, we propose to identify and characterize differential phosphorylation and interacting partners of activated STING to further understand the mechanisms by which activated STING causes BCR degradation and apoptosis in CLL. Since our preliminary data show that STING deficiency can lead to increased levels of the BCR and BCR signaling in mouse CLL, and that mouse and human malignant CLL cells significantly downregulate their expression levels of STING, we will test whether STING- downregulated or STING-deficient CLL cells are less sensitive to BTKi. We will also examine how such altered STING expression and phosphorylation can regulate the survival, progression and chemoresistance of CLL. Based on our hypothesis that activation of STING can lead to the degradation of the BCR and render CLL cells more sensitive to BTKi, we propose to combine STING agonists with BTKi to treat CLL.

NIH Research Projects · FY 2026 · 2023-01

Neurogenic overactive bladder (NOAB), characterized by urinary frequency, urgency or urgency incontinence symptoms occurring during the storage phase of the bladder, is the most common urinary complaint in multiple sclerosis (MS). Current management options for NOAB in MS have limited efficacy and considerable adverse effects, which underscores the significance of our study and highlights the need for better, less invasive therapies. Our novel study investigates brain therapeutic targets that could shift the focus of NOAB management in MS from a bladder-centric focus to brain restoration; specifically modulating the brain regions identified in our prior functional magnetic resonance imagining studies. Repetitive transcranial magnetic stimulation (rTMS) is a noninvasive brain stimulation that can modulate neurons (excite or inhibit) to improve the connectivity of the regions of interest (ROI). Our preliminary data demonstrate, for the first time, significant improvement in bladder symptoms in ten women with MS who have voiding dysfunction following multifocal transcranial magnetic stimulation without any treatment-related adverse effects. This randomized double-blind, sham-controlled single center clinical trial with an optional open-label extension (OLE) phase is designed to evaluate the effects of targeted rTMS in women with MS and NOAB by investigating restorative reorganization of brain function and improvement of urinary frequency, urgency and incontinence. We hypothesize that cortical alterations in bladder volume sensing and their response to stimulation contribute to NOAB symptoms in MS, and that improving the response to bladder distention (ROI within circuits 1 and 2) with neuronavigated rTMS can restore brain activity and improve symptoms (frequency, urgency, and incontinence). We will test our hypothesis with thses specific aims: Aim 1: To determine the clinical effects of neuronavigated and multifocal active/sham rTMS in women with MS and NOAB; Aim 2: To assess the neuroimaging restorative effects of neuronavigated active/sham rTMS in women with MS and NOAB; Aim 3: To assess the long-term safety and therapeutic effects of repeated rTMS in women with MS and NOAB who participate in the OLE phase (which subjects from both groups will be invited to enter at the 3-month follow up). Efforts to improve our current knowledge of brain contribution to lower urinary tract function and the development of an individualized, noninvasive, and effective treatment modality at the level of the brain will greatly impact the quality of life for individuals with MS and subsequently others with OAB, whether neurogenic or non-neurogenic.

NIH Research Projects · FY 2025 · 2023-01

Project Summary/Abstract Alzheimer’s disease (AD) gene-risk data, epidemiological findings and animal models converge to indicate the critical role of inflammation for the onset and progression of AD. Inflammation could provide a new focus for therapeutic intervention. However, inflammation biomarkers are poorly characterized in AD. In this project, we will measure blood and cerebrospinal fluid (CSF) inflammation biomarkers and compare them to measurements of brain glial activation obtained by positron emission tomography (PET). In addition, we will determine the effect of low-dose interleukin-2 (IL-2) immunotherapy, given over 22 weeks, on these inflammation biomarkers. For this purpose, we will measure these biomarkers in AD individuals enrolled in a phase 2a safety and efficacy trial of low-dose IL-2 therapy. Regulatory T cells (Tregs) play a neuroprotective role by suppressing inflammation in the blood and the brain. We have shown that Treg immunomodulatory mechanisms are compromised in AD patients, resulting in activation of pro-inflammatory monocytes and upregulation of inflammatory mediators. In a Phase 1 clinical study, we showed that IL-2 administration induced Treg expansion and restoration; the therapy was safe, and it was associated with improved cognition. Supported by a “Part-the-Cloud” grant from the Alzheimer’s Association, we will conduct a follow up phase 2a study which includes the measurement of cognition, as well as CSF T-tau, P-tau, Aβ42, and neurofilament light chain (NFL) before and after 22 weeks of IL-2 treatment in the subjects with mild to moderate AD. Taking advantage of this novel trial, and by evaluating the 40 AD patients participating in it, the current study will investigate systemic and central nervous system (CNS) biomarkers of inflammation and their modulation by IL-2 administration. We will analyze the impact of IL-2 immunotherapy on peripheral immune biomarkers (Aim 1) as well as CNS inflammation, measured through CSF inflammation biomarkers (Aim 2) and brain inflammation positron emission tomography (Aim 3). To determine which blood biomarkers correlate best with central nervous system biomarkers (Aim 4), the associations among blood, CSF, and brain imaging measures of neuroinflammation before and after IL-2 therapy will be determined. Our novel approach will explore the potential link between systemic and CNS inflammation in AD clinical setting and advance the use of inflammatory biomarkers in AD anti- inflammatory clinical trials.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY-ABSTRACT Astrocytes are highly-abundant cells in the nervous system and they play a critical role in the orchestration of neuronal synaptic networks. One of the primary mechanisms through which they influence neuronal synapses is thought to involve signaling via a diverse milieu of extracellular proteins (i.e., the astrocyte matrisome). However, it remains unclear which components of the astrocyte matrisome are necessary and sufficient for the formation and strengthening of neuronal excitatory synapses, especially in human-specific cells due to current limitations in experimentally investigating human neural networks using traditional monolayer cultures and immature organoids. To overcome these technical limitations, we will utilize our recently optimized approach which generates and analyzes bioengineered neural organoids that are composed of specific numbers of post-mitotic astrocytes as well as neurons that are directly transdifferentiated from human pluripotent stem cells. Specifically, we will use these bioengineered neural organoids to test the hypothesis that mature human astrocytes produce a cell type-restricted, multi-component, activity-dependent matrisome that accelerates the formation and function of neuronal synaptic networks. Our preliminary data confirms feasibility of our organoid- based approach to test this hypothesis and has identified top candidate proteins potentially underlying the astrocyte-to-neuron influence on synapses. In Aim 1, we will determine whether human astrocyte Thrombospondin 1 protein is a sufficient and necessary inducer of human neuronal synapses. We will use a combination of genetic engineering and drug treatment to conclude whether Thrombospondin 1 promotes structural and function excitatory synaptic networks and if it acts through signaling to the neuronal alpha2delta- 1 receptor. In Aim 2, we will define the extracellular human astrocyte matrisome and test its influence upon neuronal synapse formation using a novel indirect coculture approach. Cocultures will be enabled by cellular encapsulation within alginate hydrogel capsules to elucidate the extracellular astrocyte matrisome components, test their effect upon neuronal organoids, and identify relevant receptor-ligand pairs. We will investigate whether astrocyte-derived extracellular Thrombospondin 1 and/or chondroitin sulfate proteoglycans influences synapse formation and function. Finally, in Aim 3, we will test how neuronal activity influences the synapse-promoting characteristics of the human astrocyte matrisome. Neurons will be activated and paced using optogenetic tools, and the resultant effect on cocultured astrocytes will be determined using a combination of single cell RNAsequencing, pharmacological treatment, protein assays, and calcium imaging. The completion of our aims will deliver novel experimental cell lines and protocols to the scientific community, and may identify novel approaches to accelerate neuronal synaptic network formation in organoid-based model systems. Broadly, we expect our studies to make significant contributions to the neurobiology field by identifying and defining intercellular signaling mechanisms between human astrocytes and neuronal synapses.

NIH Research Projects · FY 2025 · 2022-09

OVERALL PROJECT SUMMARY Bladder cancer (BC) is the second most common urologic malignancy affecting 573,278 people worldwide in 2020. Pathologically, BC is diagnosed as non-muscle-invasive (NMI) and muscle-invasive (MI) disease. Here we define early bladder lesions as NMIBC. Major clinical gaps in NMIBC include i) lack of mechanistic insights defining NMIBC progression, and ii) lack of platform for risk stratification of NMIBC that recur but never progress (“non-progressors”), from those that progresses into MIBC (“progressors”) and consequently demonstrate poor prognosis. The goal of our Center is to tackle this clinical issue by deciphering the underlying mechanisms restraining or promoting the progression of early lesions (Project 1 & 2), and to leverage this novel biology as candidate biomarkers to risk-stratify aggressive NMIBC (Project 3). This proposal seeks to shift the current research paradigm in the field of NMIBC, by proposing a conceptually innovative tug-of-war between a tumor- restraining (Project 1) and a tumor-promoting mechanism (Project 2) in determining the outcome of early bladder lesions/NMIBC in becoming “progressors” or “non-progressors” (Project 3). Clinically, why “non-progressors” often recur but seldom progress, and what are the driving forces advancing “progressors” into MIBC with poor survival remain fundamental questions in field. Our tug-of-war hypothesis with two opposing forces is conceptually different to most other studies, which primarily focus on one side of the coin. Further, the integration of knowledge from Project 1 and 2 as a unified spatial proteomics and transcriptomics map by the Shared Resource Core will reveal spatial and temporal relationships between distinct fibroblast populations with opposing functions, their physical interactions with tumor and immune cell clusters, as well as their relationship to the biomarkers from Project 3. Benchmark of success: The knowledge gained here will shift clinical practice paradigm, by informing future NIMBC management through 1) the development of novel urinary profiling strategies that could risk stratify aggressive NMIBC (Project 1-3); 2) the identification of targets for future precision intervention, either by enhancing/sustaining the tumor-restraining mechanisms (Project 1) and/or inhibiting the tumor-promoting mechanisms (Project 2). The overall success of our program is further ensured by an extraordinary multi-investigator team that integrates three “organ-specific” bladder cancer investigators within Cedars-Sinai Medical Center. All have active R01s and individual NCI-funding track record in performing basic science research, translational bladder cancer research, or leading multi-center clinical trials on the discovery and validation of biomarkers. Finally, they propose to collect valuable retrospective and prospective NMIBC cohorts, which are essential to address the clinical questions posed within this proposal and will become available to the research community as a shared resource to advance the field.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY The goal of this academic [Houston Methodist Research Institute (HMRI) and MD Anderson Cancer Center (MDACC)] and industrial (EMPIRI) collaborative project is to validate a novel, functional cancer diagnostic assay for clinical translation to improve and personalize the care of triple-negative breast cancer (TNBC) patients. The study will validate a novel 3D tumor tissue culture method (E-slices) invented by MPI Kyuson Yun (HMRI) and licensed to EMPIRI, Inc., a biotech startup co-founded by Dr. Yun and Dave Gallup (MPI). Together with Dr. Naoto Ueno (MPI, MDACC), a renowned physician-scientist specializing in TNBC research and patient care, the team will validate the predictive accuracy of E-slices for individual TNBC patient responses to recently approved chemoimmunotherapy for early TNBC patient care. The need for personalized medicine in oncology is widely accepted but translating this important concept into clinical practice has been challenging. Currently, the dominant platform for precision medicine utilizes genomics/sequencing-based assays to measure the expression and/or mutational profiles and then infer responses to targeted therapies; however, this approach benefits <10% of patients with profiled tumors. Recognizing the inherent limitations of these inference-based methods, functional assays (e.g., organoids, PDX models) have been developed, but these approaches have numerous limitations including high cost and time required to establish the models, low “take rates”, and destruction of the native tumor microenvironment (TME). To overcome these challenges, EMPIRI developed a novel 3D ex vivo tumor culture method (E-slices) that enables rapid, personalized drug sensitivity testing in intact patient tumor tissues. E-slices retain the native TME and tissue architecture and are cultured in serum-free defined media, overcoming the limitations of other approaches. In addition, E-slices can be generated from any solid tumor and used for testing responses to chemotherapy, targeted therapy, and immunotherapy. Importantly, E-slice accuracy has been validated in a clinical setting for metastatic colorectal cancer to accurately predict individual patient treatment responses and detect inter-patient differences to the same treatments in 4-12 days, paving the way for near evidence-based personalized treatment selections. The team will: (i) determine whether E-slices predict patient responses to SOC NAC: doxorubicin (Adriamycin) plus cyclophosphamide) in a retrospective study, (ii) measure chemoimmunotherapy (Pembrolizumab plus paclitaxel + carboplatin) responses in humanized PDX slices from known responders and non-responders; and (ii) evaluate the clinical utility of E-slices in predicting TNBC patient responses to newly approved standard of care chemoimmunotherapy in a prospective study. Successful completion of this project will provide the necessary data to apply E-slices as the first functional cancer diagnostic test specifically designed to inform TNBC patient care.

NIH Research Projects · FY 2025 · 2022-09

Cell encapsulation technologies are poised to improve conventional islet transplantation to more effectively manage type I diabetes. Currently, lifelong whole-body immunosuppression is administered to avoid immune rejection of the transplant, despite the associated life-threatening adverse effects. Clinical studies reveal that transplants eventually fail due to lack of vascular support for nutrients and oxygen supply and host immune rejection. To address all these critical needs and supported by preliminary studies, we propose the NICHE, an innovative subcutaneous vascularized encapsulation system with local elution of immunosuppressants to protect transplanted cells from immune rejection. The NICHE presents dual transcutaneously refillable reservoirs, for drug and cells, respectively, separated by a nanoporous membrane. Local immunosuppressant delivery confines drugs to the graft site where immune attack occurs, minimizing exposure to the rest of the body, thus avoiding systemic immunosuppression and associated adverse effects. The NICHE cell reservoir is fully vascularized with functional vessels, recreating an ideal physiological environment conducive for maintaining long-term viability and function of transplanted cells. We hypothesize that the NICHE will provide a vascularized environment with local immunosuppressant delivery conducive for successful long-term islet allotransplantation to restore euglycemia in diabetic nonhuman primates (NHP). In aim 1, we will study the efficacy and safety of local immunosuppressant combinations as well as characterize their release from the NICHE via in vitro studies. This will be followed in aim to define the optimal local immunosuppression regimen for islet allotransplantation in NICHE and establish drug pharmacokinetics (PK) and biodistribution in healthy NHP. In aim 3, we will evaluate the curative efficacy of NICHE with allotransplanted islets to restore and maintain euglycemia in diabetic NHPs for one year. The proposed studies are based on our team’s extensive expertise in implantable drug and cell delivery systems, tissue engineering, research and clinical transplantation, transplant immunology, type 1 diabetes, as well as supportive preliminary data and previously published work. Importantly, the NICHE is designed prioritizing clinical considerations of efficacy, safety and user acceptability. Transcutaneous cell and drug refilling allow for ease of drug replenishment when needed, thus extending implant lifespan potentially for the lifetime of patients. Further, the thin and compact size of the NICHE, which is smaller than the encapsulation implants under clinical investigation, is favorable for user acceptability. Successful completion of the proposed work will provide a broadly applicable encapsulation system with localized immunosuppressant delivery for long- term protection of transplanted islets, as well as minimize adverse effects associated with immunosuppressive drugs. This could translate to a clinical breakthrough for deployment of cell therapies beyond islets, including stem cell-derived β cells, to treat diabetes as well as other diseases.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Approximately 15% of the U.S. population has chronic kidney disease, and ~700,000 patients are in full kidney failure also called end-stage kidney disease (ESKD). The optimal treatment for ESKD is living donor kidney transplantation (LDKT), followed by deceased donor kidney transplantation (DDKT); however, the standard of care continues to be ongoing dialysis, which has poor clinical outcomes in comparison to LDKT and DDKT. Best practices to transform kidney care recommended by the Centers for Medicare and Medicaid Services (CMS), the American Society of Nephrology (ASN) and the 2019 Executive Order Advancing American Kidney Health Initiative include earlier detection of patients whose kidneys are deteriorating rapidly, introducing transplant as a potential treatment option earlier, optimally before their kidneys fail, improved dissemination of health literate transplant education tools, often through digital technology or mHealth, and increasing LDKT rates by helping patients locate living donors or motivating others to donate. Barriers at the patient-, support network-, clinician- and system-levels of the Socio-Ecological Model persist, including poor identification of high-risk patients, insufficient clinician time to discuss transplant, poor transplant knowledge, reluctance or insufficient support to ask living donors to donate, and disengaged friends and relatives, some of whom who might become living donors. While extensive policy and intervention efforts are underway, none have achieved significant increases in pursuit and receipt of transplant, especially LDKT rates. In 2017, Kaiser Permanente Southern California (KPSC), an integrated care system serving 24,000 CKD patients, partnered with the Transplant Research and Education Center (TREC) at Houston Methodist Research Institute (HMRI) and J.C. Walter Jr. Transplant Center Houston Methodist Hospital (HMH) to launch a multi-year plan for transforming CKD and ESKD care. We now propose to conduct a pragmatic stepped wedge cluster randomized trial of a novel multilevel intervention to improve CKD and ESKD care, improve transplant rates and reduce disparities. One innovative component of the multilevel intervention is a state-of-the-art technology-supported Grove Kidney Health mHealth application, developed in partnership with patients, to engage patients, family members, and potential living donors to improve their CKD knowledge, view transplant success stories, and seek kidney-related support to pursue transplant, including learning how to find living donors. We also seek to identify moderators at various socio- ecological levels, especially factors influencing variations in effectiveness across different settings and among underserved patient subgroups known to have reduced access to transplant and build implementation tools to increase access to and pursuit of transplant within large integrated health systems including comparable systems (commercial, academic, safety net) across the U.S.

NIH Research Projects · FY 2025 · 2022-07

Cell encapsulation technologies are poised to improve conventional islet transplantation to more effectively manage type I diabetes. Currently, lifelong whole-body immunosuppression is administered to avoid immune rejection of the transplant, despite the associated life-threatening adverse effects. Clinical studies reveal that transplants eventually fail due to lack of vascular support for nutrients and oxygen supply and host immune rejection. To address all these critical needs and supported by preliminary studies, we propose the NICHE, an innovative subcutaneous vascularized encapsulation system with local elution of immunosuppressants to protect transplanted cells from immune rejection. The NICHE presents dual transcutaneously refillable reservoirs, for drug and cells, respectively, separated by a nanoporous membrane. Local immunosuppressant delivery confines drugs to the graft site where immune attack occurs, minimizing exposure to the rest of the body, thus avoiding systemic immunosuppression and associated adverse effects. The NICHE cell reservoir is fully vascularized with functional vessels, recreating an ideal physiological environment conducive for maintaining long-term viability and function of transplanted cells. We hypothesize that the NICHE will provide a vascularized environment with local immunosuppressant delivery for successful long-term islet engraftment to restore euglycemia in diabetic hosts. In aim 1, we will study the ability of mesenchymal stem cells to induce vascularization within the NICHE as well as modulate the NICHE immune microenvironment to be conducive for islet transplantation in diabetic rats. This will be followed in aim 2 by the biodistribution analysis of immunosuppressants locally eluted in the NICHE and the assessment of immunomodulation and pharmacokinetics in diabetic rats. In aim 3, the NICHE efficacy in providing a suitable environment for successful islet engraftment to restore euglycemia in diabetic rats will be assessed over 1 year, in parallel to a longitudinal study of NICHE microenvironment remodeling. The proposed studies are based on our team’s extensive expertise in implantable drug and cell delivery systems, tissue engineering, research and clinical transplantation, transplant immunology, type 1 diabetes, as well as supportive preliminary data and previously published work. Importantly, the NICHE is designed prioritizing clinical considerations of efficacy, safety and user acceptability. Transcutaneous cell and drug refilling allow for ease of drug replenishment when needed, thus extending implant lifespan potentially for the lifetime of patients. Further, the thin and compact size of the NICHE, which is smaller than the encapsulation implants under clinical investigation, is favorable for user acceptability. Successful completion of the proposed work will provide a broadly applicable encapsulation system with localized immunosuppressant delivery for long- term protection of transplanted islets, as well as minimize adverse effects associated with immunosuppressive drugs. This could translate to a clinical breakthrough for deployment of cell therapies beyond islets, including stem cell-derived β cells, to treat diabetes as well as other diseases.

NIH Research Projects · FY 2025 · 2022-07

Neural activity directly controls the development and homeostasis of organ function. Yet, a surprising gap exists between laboratories focused on vascular, gastrointestinal, immune, and musculoskeletal systems and those focused on neurophysiology, neuromodulation, and neural injury. Traditionally, departments are organized by organ system and this siloed structure leads to a physical and scientific jargon separation, impeding collaboration and training. Hence, our understanding of how the central nervous system communicates with end organs throughout the body is in its infancy. Neural Control of Organ Degeneration and Regeneration (NeuralCODR), a cross-disciplinary training program unites 30 faculty members and 12 clinician researchers to form mentorship teams within the largest medical research center in the world (Texas Medical Center). Mentorship teams include a primary and secondary mentor (each from one of our three main areas of research: 1) neural development and tools, 2) neural innervation and organ engineering, and 3) nervous system and peripheral organ disorders) and a clinical mentor who provides much needed and rare exposure to real-world diseases, clinical challenges, and human samples. NeuralCODR aims to: 1) catalyze the collision of talent and ideas that spawn research projects bridging neuroscience with organ systems through facilitated interactions, 2) build co-mentor teams that include neuroscience, organ systems, and clinical perspectives, ensuring trainees are guided toward a unique research niche, and 3) train fellows in research rigor, analysis, and career skills that support their development as scientific leaders. NeuralCODR leverages a network of faculty who collaborate on projects that bridge the gap between organ system biology and engineering and neural function, degeneration and regeneration, as evidenced by an impressive list of multi-laboratory shared grants, publications, and nascent collaborations. The training structure emphasizes experiences in organ engineering and organ physiology laboratories in parallel with education in neurophysiology and neural engineering, translational theory, and practice. Currently, two NeuralCODR Houston Methodist-specific postdoctoral positions exist, thanks to a philanthropic endowment that has allowed us to build the program structure and to test both the coursework and mentorship team concept. Trainees are funded for two years, during which time they have access to a core curriculum, career development and program enrichment opportunities, symposiums/retreats, and elective courses. Year one culminates with the submission of an NRSA and a manuscript based on trainees’ development of a transformative research project that incorporates non-neuroscience expertise and tools into a neuroscience problem. Year two solidifies the relationships between the mentorship team and trainee and provides training in laboratory management, didactic teaching, and mentorship skills. T32 funding will allow us to increase the current model to six postdoctoral training slots. This training program will foster a new generation of scientific leaders who pioneer research on the connected pathways between brain and organ systems to solve fundamental challenges in neuroscience.

NIH Research Projects · FY 2026 · 2022-06

ABSTRACT Metaplastic breast cancer (MpBC) is a rare subset accounting for <1% of all breast cancers. However, MpBC is a significant health challenge as it exhibits the most dismal prognosis of all breast cancers, even worse than triple-negative breast cancer (TNBC), with a survival rate of 8 months or less in patients with metastatic disease. Due to a lack of druggable targets, the main therapeutic option for metastatic MpBC remains systemic chemotherapy, despite known resistance to most cytotoxic drugs. One common molecular alteration in MpBC is hyperactivation of the phosphoinositide 3-kinase and protein kinase B (PI3K/AKT) pathway. Additionally, we recently published that MpBC displays a gain-of-function oncogenic mutation in ribosomal protein L39 (RPL39), which is responsible for treatment resistance, stem cell self-renewal, and lung metastasis. The mechanistic function of RPL39 is mediated through inducible nitric oxide synthase (iNOS)-mediated nitric oxide production. In addition, we demonstrated in a completed clinical trial that inhibiting this nitric oxide synthase (NOS) pathway using pan-NOS inhibitor NG-methyl-L-arginine acetate (L-NMMA) may represent a highly effective therapeutic option for TNBC patients. Therefore, we hypothesize that a combinatorial targeted approach of inhibiting the two major oncogenic pathways implicated in MpBC, PI3K/AKT and NOS, would lead to significant tumor regression. To test this hypothesis, this U01 application brings together research teams from Houston Methodist Cancer Center (HMCC), The University of Texas MD Anderson Cancer Center, and the National Cancer Institute (NCI). Specific Aim 1 seeks to define whether dual inhibition of PI3K/AKT using alpelisib and NOS inhibition using L-NMMA combined with nab-paclitaxel will increase the objective response rate and survival in metastatic MpBC patients. In Specific Aim 2, using blood and core biopsy tissues collected in the trial, we will identify mechanisms of response to therapy to determine the efficacy of the targeted PI3K/AKT and NOS pathway inhibitory approach. Furthermore, the cell-cell interactions among tumor cells, myeloid cells, lymphoid cells, and stromal cells within the tumor microenvironment and their role in supporting cancer stem cell populations and drug-resistant cell development during treatment will be evaluated. The impact of distinct cellular localization patterns within the tumor ecosystem on the process of cancer stem cell maintenance and modulation, as well as the development of drug resistance, will be analyzed at the single-cell level using spatial transcriptomics, immunofluorescence, CyTOF imaging systems, and a multi-modal data analysis model. This study thus proposes a mechanistic investigation of a combinatorial targeted approach against the two key pathways in MpBC, develops unique crosstalk models, and identifies biomarkers of resistance and cell–cell interactions using specimens derived from MpBC patients.

NIH Research Projects · FY 2025 · 2022-06

Modified Project Summary/Abstract Section Long-acting (LA) pre-exposure prophylactic (PrEP) strategies offer the promise of improving adherence to therapeutic regimen for maximal HIV preventive efficacy. To this end, we have developed an ultra-long acting antiretroviral (ARV) delivery implant with an unprecedented release duration of at least 2 years uninterrupted for durable and safe HIV prevention independent of user adherence. The NanoDDI is a refillable subcutaneous nanofluidic implant for constant drug release, which comprises a newly patented nanofluidic membrane, and ports for rapid, minimally invasive transcutaneous drug refilling. Refilling is performed manually via a syringe without any complex pumps or equipment to extend implant use duration beyond 2 years. The nanofluidic membrane use nanochannels to control drug release through passive diffusion without pumping mechanisms, permitting discrete, long-term user-independent dosing. Unlike injectables or other LA polymeric strategies, NanoDDI avoids burst and decay release. Zero-order release kinetics is achieved independent of physiological conditions, regardless of interindividual heterogeneity. Importantly, the NanoDDI addresses user preferences for discretion and longer dose duration. The NanoDDI development was originally planned for use with islatravir (ISL), an ARV developed by Merck. However, ISL clinical development for long-acting HIV PrEP was recently terminated by Merck based on lymphocyte depletion concerns, rendering ISL of limited clinical relevance for PrEP. Thus, we have shifted our NanoDDI focus to the potent ARVs, MK8527 (MK) and lenacapavir (LEN). Here we will test the hypothesis that constant and sustained MK8527 (MK) or lenacapavir (LEN) delivery from NanoDDI will achieve preventative drug levels for a 2-year duration and effectively prevent SHIV infection in non-human primates (NHP). This proposal outlines a comprehensive preclinical framework fundamental for developing NanoDDI as a HIV PrEP platform. We aim 1) to develop and optimize NanoDDI and MK and LEN formulations for sustained and constant release in vitro and in rats; 2) to assess pharmacokinetics (PK), tolerability, and safety of NanoDDI-MK85 and NanoDDI-LEN for 2 years in NHP and evaluate effectiveness of transcutaneous drug refilling; and 3) to comprehensively evaluate PrEP efficacy of NanoDDI-MK in NHPs using the vaginal and rectal routes of simian HIV transmission, in dose de-escalation studies. Our multidisciplinary team has a solid history of collaborative HIV PrEP studies with long-acting drug delivery implants. We will use a milestone-driven research approach to advance the NanoDDI technology towards the ultimate goal of clinical translation.

NIH Research Projects · FY 2026 · 2022-05

Cardiovascular disease is a major cause of morbidity and mortality in patients with autosomal dominant polycystic kidney disease (ADPKD). Characterized by progressive renal dysfunction, ADPKD imposes very significant healthcare and economic burdens. It has commonly been assumed that progressive renal impairment promotes cardiac disease; however, our preliminary data suggest that cardiac dysfunction originates in cardiomyocytes and manifests prior to renal failure in ADPKD. Recent clinical evidence supports our findings by showing that ADPKD patients exhibit ventricular dysfunction before the onset of renal failure, even in non-hypertensive individuals. Mutations in the gene encoding Polycystin-1 (PC1) occur in 85% of patients and are responsible for the most severe cases. Importantly, PC1 is expressed in cardiomyocytes, yet its role(s) there is(are) poorly understood. We propose that the mutant PC1 – in a cardiomyocyte-autonomous fashion – initiates and drives heart disease in ADPKD, independent of renal failure. Our data show that PC1 cardiomyocyte-specific deletion promotes systolic and diastolic dysfunction in mice. Furthermore, using a mouse model harboring a clinically established ADPKD-causing PC1 mutation (RC allele), we provide evidence of impaired calcium-cycling and contractility at the cardiomyocyte level, which occur before the onset of renal failure. Heterozygous RC/+ young mice manifest alterations in calcium handling/contractility in isolated cardiomyocytes, which correlate with reduced left ventricular global longitudinal strain and diastolic dysfunction. We discovered that PC1 regulates action potential duration via Kv channel current regulation. PC1 ablation shortens action potential duration and impairs both calcium transients and contractility in cardiomyocytes. Additionally, PC1 deletion impairs sarcoplasmic reticulum (SR) calcium loading through reduced SR calcium-ATPase (SERCA) activity. These data have led us to hypothesize that ADPKD-causing PC1 mutations disrupt PC1 actions in cardiomyocytes, impair cardiac function and predispose the heart to hypertension-induced heart failure, independent of renal dysfunction. To test this hypothesis, we propose three aims: 1) determine how PC1 mutations affect action potentials and Kv channel activity and impinge on calcium handling and contractility. 2) elucidate mechanisms whereby PC1 regulates SR calcium loading and SERCA to maintain cardiomyocyte function and test the impact of ADPKD mutations in PC1 on these events. 3) determine in vivo whether alterations in PC1 signaling in cardiomyocytes drive cardiac dysfunction and predispose the heart to hypertension-induced heart failure. Completion of our studies will provide paradigm-shifting information regarding the role of cardiomyocyte-autonomous events driving heart disease in ADPKD, the leading cause of death in these patients.