University Of Pittsburgh At Pittsburgh

NIH Research Projects · FY 2026 · 2026-05

Ovarian cancer is the most lethal gynecological cancer in the United States with poor survival due to extensive peritoneal metastasis. We discovered a previously uncharacterized early response signature to matrix detachment of high-grade serous adenocarcinoma cells, the most common ovarian cancer histologic subtype. This signature is associated with poor patient outcomes and distinct pro-metastatic signaling nodes. We find that the atypical GTPase RHOV, the most significantly induced gene in this signature, is essential for peritoneal metastasis in vivo, promoting anoikis resistance, invasion and clearance of mesothelial cells, an essential step in peritoneal metastasis. Our data reveals RHOV functions as a novel TGF-β/SMAD signaling integrator in OC, a previously uncharacterized function of this understudied GTPase. This finding represents a critical mechanistic link between early detachment responses and established pro-metastatic signaling pathways in ovarian cancer. Our overall hypothesis is that the rapid induction of the detachment response signature is a crucial step for initiating downstream signaling events necessary for ovarian cancer metastasis. Leveraging the PIs' complementary expertise in ovarian cancer research, we will use innovative approaches including in vivo CRISPR screens, auxin-inducible degradation systems, transcriptomics and imaging strategies in cell lines, xenograft tumors, and patient-derived models. In Aim1 we will identify essential genes within the clinically significant detachment response signature and delineate their convergent signaling pathways using unbiased functional genomics. In Aim 2 we will determine how RHOV and the detachment signature reprograms endocytic trafficking to change cellular signaling as exemplified by the TGF-β pathway and evaluate RHOV expression and the detachment signature as a biomarker for TGF-β inhibitor sensitivity. Defining the essential genes and downstream signaling of the detachment response including the novel RHOV- TGF-β/SMAD axis will identify key adaptations of metastatic ovarian cancer that can be targeted therapeutically. This work shifts the paradigm from studying late-stage adaptations to understanding the earliest events in the metastatic cascade, potentially leading to novel biomarker-guided precision medicine approaches for ovarian cancer patients.

NIH Research Projects · FY 2026 · 2026-05

Project summary/abstract: How the immune system maintains a balance between containing the intestinal microbiota and limiting inflammation is an important field of study with relevance to numerous human diseases. The immune system must also balance the need for having a diverse B cell receptor (BCR) repertoire with maintaining self-tolerance. Tolerance of self-reactive B cells is enforced during B cell maturation in the bone marrow (BM) and peripherally through B cell anergy, which limits their lifespan and makes them hyporeactive. Commensal microbes add complexity to the idea of tolerance, as they are an important source of foreign antigen, but are not necessarily harmful. It is rarely considered whether the continuous presence of commensal-derived antigen could have tolerogenic effects on B cells. This proposal describes a new model where we find that a microbial antigen specifically tolerizes B cells, and will test the ability of the tolerized microbial antigen-specific B cells to respond to commensal antigen or infection. We have generated three BCR-knock-in (KI) mouse strains expressing lipopolysaccharide (LPS)-specific BCRs of either low- or high- affinity heavy chains, or a high-affinity light chain, collectively referred to as 33a KIs. These BCRs were derived from an expanded B cell clone that was responding to infection with Salmonella, but also binds a portion of small-intestinal commensal microbes. 33a KI B cells show hallmarks of self-tolerance mechanisms including clonal deletion, receptor editing, and anergy. Critically, transferring BM from SPF mice into lethally irradiated germ-free mice improved B cell development, demonstrating microbially derived antigen can contribute to B cell tolerance. Strict B cell tolerance could generate immunological blind spots for certain microbial antigens, while conversely a lack of tolerance could cause overwhelming inflammation at barrier sites. Intriguingly, we observe that 33a KI B cells form germinal centers in the Peyer’s patches (PPs) at steady-state, and a robust splenic extrafollicular response to systemic Salmonella infection, demonstrating that despite tolerization, the B cells can respond in certain contexts. The central goal of this proposal is to define the fundamental properties of this novel type of B cell response during homeostasis and following infection. In Aim 1 we will determine whether 33a B cells undergo classical germinal center responses in the PP during homeostasis, including the formation of long-lived memory B cells and plasma cells. In Aim 2, we will define the 33a B cell response to infection, and test whether the nature of antigenic stimulation impacts their response using oral infection with mutant Salmonella strains and a closely related pathogen, Yersinia pseudotuberculosis. This project tests a new conceptual framework that microbial antigen can contribute to both B cell tolerance and inflammation as needed, which will in turn have important implications for understanding many human disease conditions. The knowledge and skills I will attain from the proposed work will be broadly applicable and help me achieve my overall goal of becoming an independent investigator studying the immune balance of tolerance and protection.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Across species, reproduction and immunity compete for limited physiological resources—a trade-off that becomes increasingly consequential with age, as both reproductive capacity and immune competence decline. In mammals, proteins that mediate these relationships to coordinate reproductive investment, immune regulation, and somatic maintenance are scarcely defined. This presents a critical gap in our understanding of reproductive aging and age-related disease susceptibility. This proposal investigates whether the C. elegans protein TCER-1, an established regulator of the fertility–immunity–longevity axis, has a conserved functional homolog in mammals. We previously identified TCER-1 as a pro-longevity factor that maintains fertility with age by suppressing immune and stress responses. Its homolog in Arabidopsis also promotes fertility and represses immunity, and our collaborative studies reveal a similar role for the Drosophila TCER-1 in repressing stress resistance, highlighting conserved trade-off regulation across kingdoms. TCER-1 is homologous to the mammalian transcription elongation and splicing factor, TCERG1, which has been linked to human disease in GWAS studies, though it’s in vivo function remains uncharacterized. This proposal is based on our preliminary data showing that (i) worm and mammalian TCERG1 both regulate alternative splicing of immune-related genes, and importantly (ii) mouse and human TCERG1 are enriched in germ-cells and exhibit a major age-related decline in female oocytes. In this exploratory proposal, we aim to assess whether human or mouse TCERG1 can functionally replace TCER-1 in C. elegans (Aim 1) and to define the role of mouse TCERG1 in fertility, ovarian reserve and reproductive aging by using existing and newly generated knockout models (Aim 2). By uncovering a potentially conserved molecular regulator of physiological trade-offs, this project aims to illuminate how reproductive aging shapes immune function and broader somatic aging, and to lay the groundwork for future interventions that support reproductive health in aging individuals.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT Acute lung injury (ALI) and its severe form, acute respiratory distress syndrome (ARDS), are life-threatening conditions that result from direct or indirect lung damage, leading to uncontrolled inflammation, alveolar injury, and impaired oxygen exchange and impaired lung repair. Despite advancements in clinical care, including lung- protective ventilation and corticosteroids, ARDS remains a major cause of morbidity and mortality, with no targeted pharmacological treatments available. Resolution and restoration of lung function require clearing pro- debris and inflammatory cells while repairing the alveolar-capillary membrane. Effective therapies should both suppress inflammation and promote lung repair to prevent ARDS progression and reduce long-term sequalae. Regulatory T cells (Tregs) are critical in maintaining homeostasis by suppressing other immune cells. Recently Tregs have emerged as reparative cells when stimulated by IL-33 via the IL-33 receptor, ST2. Unfortunately, lung injury leading to ARDS seems to overwhelm endogenous Treg responses and the lengthy manufacturing process of expanding autologous Tregs limits their applicability as an effective therapy for acute lung injuries. My preliminary data demonstrate that IL-33 stimulates expression of ST2, while IL-2 alone promoted secretion of the IL-33 antagonist soluble ST2 (sST2). Both allogenic and syngeneic IL-33-stimulated Treg were protective against ALI induced mortality when delivered 24 hours post lung injury and promoted the restoration of alveolar macrophages in the lung interstitium. This was in contrast to the suppressive IL-2 simulated Treg group that limited alveolar macrophage frequency. Therefore, my hypothesis is that limiting Treg sST2 expression, while augmenting expression of ST2 will enhance Treg reparative responses and improve the effectiveness of Treg cell therapy for ALI. I will directly test this hypothesis by addressing the following Specific Aims: 1. Define how Treg regulate expression of ST2 isoforms. 2. Determine the capacity of IL-33-programmed syngeneic (syn) or allogeneic (allo) Treg adoptive cell therapy (ACT) to promote tissue repair and resolution after ALI. Completion of these aims will establish how Treg regulate ST2/sST2 expression to balance repair and suppression. I will also determine whether IL-33 stimulated Tregs or Tregs engineered to have sustained ST2 expression in the absence of sST2 are effective against ALI mortality. These studies will also determine if allogenic Treg are as effective as autologously sourced cells and address mechanisms by which Treg modulate alveolar macrophage re-establishment in the lung. Contribution to Training: This proposal entails an extensive training plan to elucidate immunoregulatory and reparative functions of Treg combined with superb mentorship in bioinformatics, immunology, and cell therapy. Together, the generated data will foster my professional development for conference talks and manuscript preparation. It further integrates my interests in developing clinical therapies aimed at mitigating immune-mediated damage after ALI and provides strong foundations for an industry career by collaborating with renowned researchers in transplantation, regenerative medicine, and infectious disease.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT This proposal aims to unveil the mechanisms by which graft-derived extracellular vesicles (EVs), carrying donor antigens (Ags) and immunoregulatory mediators, initiate the innate and adaptive immune responses that cause rejection of allografts. Transplant rejection remains a critical barrier to long-term graft survival, with chronic rejection affecting 50-80% of heart transplant recipients within five years. Current treatments, which broadly suppress the immune system, carry risks of infections, malignancies, and organ toxicity, while failing to address donor-specific immune activation—such as lingering anti-donor T cells inflicting persistent damage on allografts. Nonetheless, transplantation remains a life-saving intervention for end-stage organ failure. Recent findings by our group and others have challenged the established paradigm of allorecognition, which posited that donor antigen-presenting cells (APCs) mobilized from organ or tissue allografts to the graft-draining (d) secondary lymphoid tissues (SLTs) are the main initiators of anti-donor T- and B-cell alloimmunity. Our preliminary data reveal that graft-derived EVs, not donor APCs, represent the earliest platform by which allogeneic (allo)-Ags are relayed to the recipient’s APCs in SLTs. The family of EVs includes vesicles with different sizes, biogenesis, and composition (e.g. exosomes, microvesicles, etc) that facilitate local and systemic signaling by transferring proteins, Ags, immunoregulatory mediators, RNAs, and lipids between cells. During the past decade, it has been established that EVs are central mediators of horizontal cell-to-cell communication, although their functions in vivo remain largely unknown. This gap in knowledge and our preliminary findings on the role of EVs in allorecognition in transplantation led to my central hypothesis that “graft-derived EVs initiate the anti- donor immune response in recipient SLTs.” This application will investigate the role of graft-EVs in allorecognition in situ and in vivo using transplant models in mice. I will test my hypothesis with two aims: Aim 1 will investigate the cellular sources and cargo of graft EVs using skin and heart allografts from mice genetically engineered for in vivo tracking of fluoroprobe-tagged graft- EVs and their respective parent cells using two photon microscopy. This will be done in combination with proteomics analysis of the graft-EVs. Aim 2 will test if allo-EVs released under proinflammatory conditions activate and affect the phenotype, transcriptome, and function of the acceptor APCs (dendritic cells) in dSLTs (Aim 2A) and compare the ability of allo-EVs to activate naïve vs. memory T cells in SLTs (Aim 2B). My long-term goal is to understand how graft EVs function in vivo to provide new grounds for the development of EV-based therapies and disease markers of clinical relevance to transplantation and immune-mediated disorders. This fellowship will allow me to gain expertise in transplant immunology and the biology of EVs, gain proficiency in computational analysis in the fields of proteomics and transcriptomics, and acquire research mentorship tailored to my career goal of becoming a physician-scientist in the field of transplant immunology.

NIH Research Projects · FY 2026 · 2026-05

Project Summary / Abstract Despite numerous improvements in treatment and prevention, cardiovascular disease (CVD) remains one of the leading causes of morbidity and mortality worldwide. Nutrients such as lipid, particularly derived from the diet, have long been linked to the development and exacerbation of CVD. However, the role of other nutrients such as amino acids in the pathogenesis of CVD has largely remained unexplored. In work spanning the past decade, we have implicated amino acids as key stimuli in pathogenic mTOR signaling in macrophages via a lysosomal nutrient-sensing mechanism. We further identified leucine as the single most important nutrient driving this lysosomal mTOR signaling in atherosclerosis. A significant contributing factor to leucine-mediated signaling is the concomitant increase in a major leucine transporter, SLC7a5/LAT1. Macrophage Slc7a5-deficiency significantly reduces mTOR signaling and plaque size and complexity in atherogenic mice. We have also leveraged the upregulation of leucine transport as a diagnostic approach to imaging atherosclerotic plaques by demonstrating the affinity of a leucine-like PET radiotracer to macrophage-rich areas of lesions. Beyond leucine uptake, we recently discovered that its catabolism is inhibited in plaque macrophages which exacerbates leucine accumulation and mTOR. Overall, we provide multiple promising approaches to modulating leucine-mTOR signaling in macrophages. In this proposal, we will impinge on each of these processes by genetic and pharmacological means in order to test the hypothesis that perturbation of leucine uptake, lysosomal sensing, and metabolism can be effective diagnostic and therapeutic strategies in atherosclerosis. This will be achieved in three distinct aims: 1) to evaluate the diagnostic and therapeutic potential of modulating macrophage leucine uptake in atherosclerosis, 2) to dissect the salient leucine-sensing mechanisms at the lysosome in driving mTOR activation and atherosclerosis, and 3) to evaluate the therapeutic potential of enhancing macrophage leucine catabolism in atherosclerosis. We will use an innovative set of tools including novel PET probes and a diverse array of mouse models to pursue a highly mechanistic evaluation of macrophage leucine-mTOR signaling in CVD.

NIH Research Projects · FY 2026 · 2026-05

Koob and colleagues have postulated that an imbalance between neurotransmitters in the brain stress and anti-stress systems drives negative reinforcement and compulsive alcohol use in alcohol use disorder (AUD). Nociceptin (N/OFQ), which binds to the nociceptive opioid peptide receptors (NOP), is one such neurotransmitter in the brain that regulates stress and resilience in animal models of addiction. N/OFQ, when infused in the brain, increases corticosterone, adrenocorticotropic hormone, and corticotrophin- releasing factor (CRF), all components of the hypothalamic-pituitary-adrenal axis that regulate stress responses. CRF infusions have also been shown to upregulate NOP, presumably to enhance N/OFQ signaling, in brain regions that regulate stress. Surprisingly, both NOP agonists and antagonists show therapeutic effects in rodent models of AUD. However, rodent models do not clarify which, if any, subtype of AUD subjects (e.g., heavy drinking) will benefit from treatment with NOP agonists Vs. antagonists. Previous [11C]NOP-1A PET studies conducted by our group in humans with AUD have demonstrated lower binding (VT) to NOP receptors in heavy relative to nonheavy drinking AUD subjects. In these AUD PET studies, lower NOP receptors also predicted relapse to alcohol in a 12-week contingency management protocol that incentivized subjects with money to abstain. Assuming lower NOP receptors reflect higher N/OFQ levels; these results suggest that increased N/OFQ signaling in heavy drinkers contributes to their inability to abstain during treatment. Blocking excessive N/OFQ signaling with a NOP antagonist drug (LY2940094) has also been shown to decrease heavy drinking days and increase abstinent days in AUD subjects in clinical trials. An emerging view in the field is that a hyperactive N/OFQ-NOP receptor system, in response to increases in CRF transmission, induces hyperkatifeia, negative reinforcement, and relapse in AUD. To investigate CRF X NOP interactions in humans, we designed a PET experiment to measure [11C]NOP-1A VT before and after an intravenous (IV) hydrocortisone challenge in healthy controls (HC). Hydrocortisone administration led to a 10 to 15% increase in [11C]NOP-1A VT in brain regions, including the amygdala. Increased NOP measured in response to cortisol, and by extension, CRF, in this paradigm reflects an individual’s ability to enhance N/OFQ transmission during stress. Here, we propose to use this novel imaging paradigm to compare hydrocortisone-induced increases in [11C]NOP-1A binding (DVT) in the amygdala (and secondary reward regions) in heavy drinking AUD subjects Vs. HC. We hypothesize that hydrocortisone-induced increases in [11C]NOP-1A binding (DVT) will be larger in heavy drinking AUD relative to HC (aim 1), and this will predict relapse to alcohol (aim 2). Such a result will support the presence of a hyperactive NOP receptor system in response to increases in cortisol/CRF during conditions such as stress, chronic pain, etc., promoting relapse in heavy drinking AUD subjects.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Cell cycle dysregulation is a fundamental driver of cancer progression, yet our understanding of cell cycle regulation remains largely based on a singular, canonical model. Recent technological advances have revealed that cells can execute distinct cell cycle programs - unique configurations of molecular networks that emerge from genetic and microenvironmental inputs. However, the prevalence and consequence of this cell cycle plasticity in cancer remain poorly understood. This study introduces cell cycle plasticity as a novel conceptual framework to understand bladder cancer heterogeneity. Using innovative single-cell spatial proteomics and machine learning approaches, we will map cell cycle programs across 661 bladder tumors spanning diverse histological and molecular subtypes. We hypothesize that bladder cancer cells execute distinct cell cycle programs that contribute to tumor progression and are shaped by the tumor microenvironment (TME). Through three integrated aims, we will: (1) catalog cell cycle programs driving bladder cancer growth and their association with histological/molecular subtypes, (2) investigate how specific TME configurations influence cell cycles using graph neural networks, and (3) develop experimental models that recapitulate cell cycle programs for mechanistic studies. This work will generate the first systematic catalog of cell cycle heterogeneity in bladder cancer and establish a publicly available resource of single-cell proteomic data mapping cell cycle effectors and TME biomarkers. Beyond advancing our fundamental understanding of bladder cancer biology, this study will provide a foundation for future biomarker discovery and therapeutic targeting strategies aimed at improving patient outcomes through precision oncology.

NIH Research Projects · FY 2026 · 2026-05

Patients with diabetes mellitus (type 1 or 2) have a total lifetime risk of a diabetic foot ulcer (DFU) complication as high as 25%; 14-24% of them suffer from amputation. People with T1D develop DFU at a younger age and are at a much greater risk of amputation and hospitalization secondary to a DFU compared to T2D. The difference between pathophysiology and outcomes for individuals with T1D versus T2D is poorly understood and understudied. Family and twin-based studies have identified significant genetic components especially single nucleotide variations (SNV) in T1D as compared to T2D subjects. However, systematic patient-based genetic studies of T1D DFU are scanty, and the proposed work is aimed at seeding a novel paradigm in wound healing research. The originality and strength of our study stems from the genome-wide genotyping feasibility studies on robust quality controlled and parametrically qualified genotyped data of 149 chronic wound patients with diabetes status. This study identified 20576 SNV significantly associated with human chronic wounds (p- value<0.01, CR>97%, MAF>0.01). Majority (>60%) of these SNP were predicted to be causative for truncated or nonfunctional proteins using Variant Effector Prediction analysis were identified. To investigate the clinical significance of wound associated SNV, a meta-analysis against the phenotypes annotated in GWAS catalog was conducted as reported. These SNVs were intersected with manually curated >270,000 GWAS SNPs annotated with ~900 GWAS phenotypes collected from ~2500 studies. Enrichment analysis of the above intersected SNVs was performed against these GWAS phenotypes and respective odds ratio, and level of significance were calculated using Fisher’s exact test. These analyses identified “obesity” as the most significantly enriched GWAS-phenotype (log2 odds ratio = 4.06, p-value= 4.94E-12) for wound associated SNPs predominantly present in fat mass and obesity-associated (FTO) gene. This proposal is responsive to RFA-DK-26-009 for the New Investigator Gateway Award for collaborative type 1 diabetes (T1D) Research through Diabetic Foot Consortium (DFC). The objective of the proposed work is to determine SNV T1D and T2D that contribute to diabetic wound closure. This study will investigate the wound tissue already collected from patients with open DFU (N=50 with T1D and n=100 T2D) enrolled in the DFC Master Protocol. The following specific aims are proposed: 1.0 Aim 1. Identify SNV uniquely associated with T1D non-healing phenotype. T1D vs T2D will identify T1D-specific SNV (SNVT1D). Healing vs non-healing will identify SNVT1D-NH. SNVT1D-NH will be shortlisted to obtain candidate SNVT1D- NH (cSNVT1D-NH) based on overlap with obesity-associated SNP. 2.0 Aim 2. Test the functional significance of cSNVT1D-NH in wound healing mechanisms in vitro. Gene editing to specifically induce risk to non-risk alleles of specific cSNVT1D-NH using CRISPR/Cas9 genome editing improves: 2.1 epidermal keratinocyte migration in an in vitro scratch model; 2.2 formation of well-perfused and non-leaky vessels by microvascular endothelial cells using 3D-angiogenesis assay; and 2.3 augmentation of collagen deposition and maturation by dermal fibroblasts.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT Suicide is the second leading cause of death among young adults ages 15-241, with rates continuing to rise2. While research has identified some broad predictive factors3, our ability to predict who will experience suicidal thoughts and behaviors (STB) and when these crises will occur remains limited4. This challenge stems from the fact that suicide risk fluctuates dramatically in response to emotional and interpersonal distress5,6, with social threats often acting as precipitating events7,8. The tendency to respond impulsively to negative emotions (e.g., negative urgency9) may help explain why some individuals engage in STB as a maladaptive attempt to escape emotional pain following social threat or rejection. Evidence from neuroscience indicates that social- affective circuitry reflects subjective affective sensitivity to social threat10,11, and overlaps with putative neural correlates of negative urgency12,13, suggesting a potential neural profile that may drive associations between social threat and STB. To test this, I will utilize data from an ongoing R01 including 6 months of ecological momentary assessment (EMA), a personalized peer social feedback fMRI task, and self-report questionnaires from 150 young adults (ages 18-30) with chronic STB to examine how function in social-affective systems and real-world experiences of social threat interact to predict STB. The Specific Aims of this study are to: (1) test associations between negative urgency and STB using both baseline and prospective EMA assessments; (2) investigate associations between functional connectivity of social-affective systems during social threat and trait-level negative urgency; and (3) examine whether individual differences in neural response to social threat moderate same-day relationships between social rejection-generated negative affect and suicidal thoughts. This project, and the associated F31 fellowship at the University of Pittsburgh, will provide critical training for the applicant to become an independent researcher investigating how neural and behavioral responses to social contexts influence suicide risk during key developmental periods. To accomplish the proposed research, this application includes a comprehensive training and mentorship plan that builds on the applicant’s prior clinical psychology and developmental neuroscience training. These Training Goals will focus on expanding the applicant’s knowledge and/or skills in: (1) neurodevelopmental pathways to suicide; (2) negative urgency as a mechanism of suicidal thoughts; (3) task-based fMRI methods, with an emphasis on functional connectivity analyses; and (4) implementing mixed-effects modeling for intensive longitudinal data. These goals will be accomplished through mentorship meetings, workshops, conferences, and coursework with a committed interdisciplinary team. Complemented by support from a dedicated research environment at the University of Pittsburgh, this fellowship will accelerate the applicant’s trajectory toward becoming an independent researcher focused on using multimodal research to identify how individual neurobiology interacts with one’s social environment to create enduring risk for STB.

NIH Research Projects · FY 2026 · 2026-04

Abstract: Lupus nephritis (LN) is a common and severe manifestation of systemic lupus erythematosus (SLE), which is associated with significant morbidity and mortality. Nearly 10% of patients with LN develop ESRD and require dialysis. SLE and specifically LN are highly associated with interferon (IFN) activity. While traditionally a type I IFN (IFNα/β) signature has been implicated in SLE, more recent work reveals a significant overlap in the downstream transcripts regulated by IFNα/β, type II IFN (IFNγ), and type III (IFNλ) signaling. Studies in humans and mice implicate each of these IFNs in SLE and LN. Our preliminary data identified a novel protective role for IFN signaling in the tissue parenchyma. Using bone marrow chimeras, deletion of hematopoietic IFNγ receptor (IFNγR) results in disease suppression, while deletion of IFNγR in the tissue parenchyma accelerates disease, indicating a dichotomous effect of IFNγ on SLE pathogenesis. We hypothesize that IFNγ and possibly type I and III IFNs initiate a tolerogenic program in the kidney by either downregulating inflammatory pathways or inducing suppressive molecules. Given this novel finding, we plan to answer several fundamental questions regarding this process. - Do type I and/or type III IFN also stimulate a similar protective program in the kidney? We hypothesize that all IFNs will also induce tissue-protective effects. However, as protection is a dominant feature, we expect the mechanisms by which type I, II, and III IFNs provide this protection in the parenchyma would differ. - What are the mechanisms by which IFNγ mediates protective effects in tissue parenchyma? RNAseq and validation studies suggest several possible mechanisms by which IFNγR-induced signaling in the tissue parenchyma regulates disease. This proposal will explore mediators of this protective effect. - How does the tissue microenvironment alter immune cells infiltrating the tissue? Previously, we identified a suppressed/dysfunctional phenotype of kidney-infiltrating T cells. We hypothesize there is a unique interaction between infiltrating immune cells and the tissue microenvironment. Therefore, we hypothesize that in tissue that cannot respond to immune stimulation by IFNs and/or other cytokines, there may be distinct functional and transcriptional changes of tissue-infiltrating immune cells. We propose to address the above questions by employing several methods to assess parenchymal signaling, including two distinct murine models of SLE, bone marrow chimeras, and Cre-lox systems. Indeed, we propose creating the first kidney-specific Cre on a lupus-prone mouse strain and evaluating a novel concept of immune regulation by the kidney parenchyma. Not only will this proposed project identify unique roles for parenchymal response to immune signaling, but it will also provide a window into how infiltrating immune cells are altered by the tissue microenvironment. This will transform how we think about lupus nephritis and numerous other kidney diseases, many of which have an underappreciated immune-mediated component.

NIH Research Projects · FY 2026 · 2026-04

SUMMARY – Overall The overall objective of this application is to develop bacteriophage (phage) therapy as a new medical countermeasure against multidrug-resistant (MDR) Pseudomonas aeruginosa infections. Antibiotic resistance is an urgent global health threat, and MDR P. aeruginosa strains cause deadly infections like pneumonia and bacteremia. Phage therapy holds significant potential as a new therapeutic modality against MDR P. aeruginosa. Here we propose to leverage the collective expertise of well-established leaders in the phage therapy field to form the Pitt Center for Accelerating Phage Therapy (P-CAPT) Program. The P-CAPT Program will unify the efforts of phage experts at the University of Pittsburgh, University of Southern California, Walter Reed Army Institute of Research, and Intralytix, Inc. to develop methods and models that will accelerate phage therapy for MDR P. aeruginosa infections toward widespread clinical use. Through an alliance of two Research Projects supported by two Scientific Cores, the P-CAPT Program will develop rigorous assays and novel tools to optimize phage cocktail design (Research Project 1) and establish optimal phage dosing protocols using systems-based models (Research Project 2). Scientific Core 1 (Phage Core) will provide comprehensive and standardized phage development, analysis, and cGMP manufacturing for the entire P-CAPT Program. Scientific Core 2 (Animal Core) will conduct necessary in vivo testing of individual phages and phage cocktails using well- established mouse models of P. aeruginosa respiratory tract infection. Throughout the P-CAPT Program, investigators will capitalize on their prior experience to catalyze preclinical phage development targeting MDR P. aeruginosa, which will serve as a model for phage therapy research and development for all ESKAPE pathogens. The distinguishing features of this program that set it apart from others are the highly relevant expertise of the participating investigators, the use of phages that have already been administered to patients under compassionate use and in clinical trials, access to biospecimens collected from dozens of phage therapy patients, and the participation of Intralytix as an industry partner. Overall, the P-CAPT Program will advance efficacious phage therapy for MDR P. aeruginosa infections, and will contribute to the CAPT-CEP mission of enhancing the quality and efficacy of phage therapeutic products, thereby unlocking this therapy's full potential to combat ESKAPE pathogen infections.

NIH Research Projects · FY 2026 · 2026-04

ABSTRACT The U.S. recently reached a historic milestone, surpassing one million life-saving organ transplants. For these patients, tacrolimus—a calcineurin inhibitor (CNI)—is the cornerstone of immunosuppression, prescribed to over 90% of transplant recipients due to its proven efficacy and tolerability. However, tacrolimus is associated with a distinctive triad of kidney-centered adverse effects: hypertension, hyperkalemia, and metabolic acidosis. Intriguingly, this triad resembles the rare Mendelian disorder Familial Hyperkalemia and Hypertension (FHHt), which results from over-activation of the NaCl-cotransporter (NCC) in the distal convoluted tubule. NCC activity is regulated by the WNK/SPAK (With-no-lysine and Ste20/ SPS-1 -related- proline- alanine- rich protein kinase) signaling pathway. In the distal convoluted tubule, kidney-specific WNK1 (KS-WNK1) is essential for activation of the WNK/SPAK/NCC cascade. Mice carrying human gain-of-function mutations in KS-WNK1 have over- activation of this pathway, thereby mimicking both the CNI- and FHHt-associated phenotypes. We propose that KS-WNK1 acts as a scaffold, recruiting WNK/SPAK components into biomolecular condensates known as WNK bodies, thereby amplifying NCC signaling. Biomolecular condensates are membraneless compartments that concentrate macromolecules through phase separation, creating localized environments that regulate specific biochemical reactions. The study of condensate biology has reshaped our understanding of intracellular organization and has sparked growing interest in pharmacologic strategies to target these structures. Our preliminary data indicate that tacrolimus increases the abundance of WNK bodies, suggesting a direct mechanistic link between CNIs, condensates, and NCC overactivation. The goal of this R01 is to use CNIs as both an experimental tool and clinical model to manipulate WNK body composition, dynamics, and function, while concurrently testing how CNIs alter kidney physiology through WNK bodies. Our central hypothesis is that calcineurin inhibition by tacrolimus stabilizes KS-WNK1 by increasing its phosphorylation and reducing its ubiquitylation. The resulting increase in KS-WNK1 abundance promotes WNK body assembly, amplifies WNK/SPAK/NCC signaling, and leads to the clinical triad of hypertension, hyperkalemia, and metabolic acidosis. We intend to address the hypothesis through: (1) Cellular models to define how CNIs alter KS-WNK1 post- translational modifications and WNK body dynamics; (2) Mouse models to test whether mice lacking or overexpressing WNK bodies are resistant or hypersensitive to CNI-induced side effects; and (3) Human allograft kidney biopsy samples to correlate WNK body abundance with hypertension, hyperkalemia, and acidosis. Using integrated molecular and whole-organism approaches, the expected outcomes include: (i) validation of WNK bodies as pharmacological targets of CNIs; (ii) mechanistic insight into how CNIs modulate WNK bodies to alter renal electrolyte handling; and (iii) identification of CNI-treated patients who may benefit from adjunctive therapy with thiazide diuretics to block NCC overactivation.

NIH Research Projects · FY 2026 · 2026-04

ABSTRACT This proposal aims to investigate the role of hepatocyte senescence in liver function and the development of metabolic dysfunction-associated steatotic liver disease (MASLD), a disease which affects about 33% of the global population. Cellular senescence, a state of irreversible cell cycle arrest triggered by multiple cellular stressors such as DNA damage and telomere shortening, is increasingly recognized as a contributor to metabolic dysfunction associated with aging, particularly in the liver, where it is implicated in mitochondrial dysfunction, steatosis, inflammation, and fibrosis. However, the precise role of senescent hepatocytes in MASLD remains poorly understood. Previous studies have linked senescent hepatocytes to impaired fatty acid oxidation and liver steatosis; however, these conclusions were largely based on positive associations between histological markers of senescence and stains for cellular lipids and used models with heterogeneous senescence onset. To address these limitations, we developed a novel in vivo model where we can selectively induce senescence in hepatocytes of adult mice to investigate the effects of senescence on liver function and the progression of MASLD. Our preliminary studies revealed that mice with senescent hepatocytes were protected from MASLD by the means of reduced weight gain, adiposity, and liver fat when exposed to a western diet. Moreover, these mice showed improved glucose tolerance and hepatic insulin sensitivity while maintaining mitochondrial respiratory capacity. These findings suggest that the energetic demands of senescence may confer adaptive metabolic benefits during short-term metabolic stress. Therefore, our central hypothesis is that senescent hepatocytes exhibit an increased energetic burden to maintain homeostasis, which may provide protective effects against liver fat accumulation and metabolic dysfunction during short-term dietary stress. To test our hypothesis, Aim 1 will assess cellular metabolism and energy expenditure in senescent hepatocytes both in vivo and ex vivo using our novel transgenic mouse model. Aim 2 will explore the molecular mechanisms that connect senescent hepatocytes with metabolic adaptation. To accomplish this, we will establish an in vitro system to induce DNA damage and use CRIPSR/Cas9 mediated genome editing to probe the downstream mediators of adaptation. We will also validate our in vitro finding in genetically modified mice that lack DNA damaging signal transducer CHK2. Completion of the above aims will expand our understanding of the contribution of cellular senescence to metabolic health while also providing outstanding training in advanced skills in metabolic isotope tracing, metabolic phenotyping, and molecular biology, supported by strong mentorship and career development resources.

NIH Research Projects · FY 2026 · 2026-04

Project Summary/Abstract: Sickle cell disease (SCD), a major global health burden affecting millions worldwide, is characterized by chronic anemia that drives progressive multi-organ damage through a vicious cycle of tissue hypoxia, sickling, and hemolysis. However, safe and effective therapies for anemia in SCD are limited, and recent withdrawals of SCD therapies have emphasized the importance of understanding patient-specific benefits and risks of therapies. While hemoglobin (Hb)-raising therapies such as hydroxyurea (HU), blood transfusions, and novel agents aim to alleviate anemia, their effects on hemorheology (e.g., blood viscosity, red blood cell [RBC] deformability) and tissue perfusion remain poorly understood. This knowledge gap is critical, as excessive Hb increases in SCD may increase blood viscosity and paradoxically worsen vaso-occlusion and impair tissue perfusion, particularly in HbSC disease, where higher baseline Hb levels result in higher blood viscosity. Current clinical practice lacks biomarkers to guide individualized therapy, relying instead on Hb levels—a flawed surrogate biomarker that fails to capture the interplay between anemia correction, blood flow dynamics, and tissue oxygenation. This study seeks to address existing knowledge gaps by evaluating the impact of anemia therapies on tissue perfusion and hemorheological biomarkers and identifying baseline predictors of therapeutic response in patients with SCD. The aims of this study are: 1) To determine the effects of anemia treatments (HU, transfusions) on cerebral tissue oxygenation (StO₂) and hemorheology in adults with SCD, leveraging an international cohort in Nigeria to enhance sample size and reduce treatment heterogeneity, and 2) To define how clinico-demographic factors (SCD genotype, geographic setting) influence hemorheological profiles and StO₂. Using near-infrared spectroscopy (NIRS) to measure StO₂ as a primary physiological endpoint, this study will longitudinally assess treatment effects while incorporating a comprehensive panel of hemorheological biomarkers, including whole blood viscosity, RBC deformability, and cellular adhesion, to monitor individual physiological responses. Additionally, understanding whether SCD genotype or geographic cohort influences baseline hemorheology and StO₂ will establish a foundation for appropriate comparisons and treatment strategies across diverse SCD populations and inform future international clinical trial designs. Successful completion of these aims will identify translatable biomarkers to guide personalized anemia treatment, elucidate mechanisms underlying phenotypic variability in SCD, and inform the design of a future adaptive clinical trial comparing anemia therapies. Through this career development award and guidance from my multidisciplinary mentorship team, I will acquire advanced skills in hemorheological assays, optical biomarkers (NIRS), advanced clinical trial design, Bayesian statistical analysis, and global translational research leadership and execution. This training will position me to conduct innovative and efficient trials that address the global burden of SCD, fostering my transition to independence as a physician-scientist focused on optimizing anemia management in SCD.

NIH Research Projects · FY 2026 · 2026-04

Summary/Abstract Whether we are fatigued from a teleconference call, eager to read a book, energetic and ready to start our day or tired and preparing for sleep, anxious or happy, in the real world key neurocognitive processes and moods fluctuate over minutes-to-hours. Hunger, circadian rhythms, hormone levels, blood sugar, heart rate, body temperature, all fluctuate at the mesoscopic timescale of minutes-to-hours modulating how we respond to the world. For example, the response of the brain to the odor of fresh cookies just out of the oven is different when one is hungry versus for someone who just ate. These state fluctuations also influence why the same event will trigger pathological symptoms at some times, but not others, in individuals with neurological or psychiatric disorders. Despite the critical importance of interrelated brain, cognitive, behavioral, and physiological fluctuations at the timescale of minutes-to-hours in the real world, nearly everything we know about human brain function mostly comes from highly controlled experiments lasting milliseconds-to-seconds, or longitudinal studies from months-to-years. This has left a critical gap in scientific knowledge regarding the dynamics of brain, cognitive, behavioral, and physiological interactions at the timescale of minutes-to-hours in real world settings. To fill this gap, we will develop a platform for measuring and analyzing brain activity, cognitive states, behavior, and physiology leveraging the unique opportunity to record multiday neural activity during natural behavior in the real world in individuals undergoing the surgical treatment for epilepsy. We will integrate cutting- edge wearable and artificial intelligence technology for rich behavioral and physiological monitoring along with neural network and dynamical systems learning approaches. We will rigorously quality control and validate this platform to ensure the highest quality data. We will also perform a bioethics study to develop a paradigm that addresses the unique privacy concerns that arise in using ubiquitous measurements for real-world neuroscience in this participant population. In the R33 phase of this project, we will then use this platform to examine how brain, cognitive, behavioral, and physiological interrelationships fluctuate with circadian rhythms and relate to brain network excitability. Given the ubiquity of circadian rhythms across the brain and body, their influence on cognition and behavior, and their relationship with many neurological and psychiatric disorders, the R33 study has broad implications for both our understanding of the brain and for potential treatment strategies for many disorders. For example, knowing when brain networks are in high or low excitability states can suggest windows of opportunity for maximally beneficial clinical interventions. The development of this integrated brain, cognitive, behavioral, and physiological measurement and analysis platform in the R61 phase of this project will open up new neuroscientific and translational research opportunities. The studies on circadian rhythms and brain network excitability in the R33 phase will lead to new understanding of these important neurobiological processes and provides insights with broad clinical and translational implications.

NIH Research Projects · FY 2026 · 2026-04

The evolving geopolitical landscape has heightened the risk of radiation-induced skin injuries due to large- scale radiological incidents, including state or terrorist-improvised nuclear devices. Additionally, radiation induced skin injuries are a significant adverse effect of radiotherapy in cancer patients. However, effective countermeasures remain limited, primarily due to the lack of physiologically and anatomically relevant human models for studying radiation-induced injuries.This proposal aims to address this critical gap by developing an extracorporeal human skin perfusion model to investigate radiation injury mechanisms and facilitate the development of effective therapeutics. The long-term goal is to create a fully automated, immune- competent human skin perfusion system that mimics in vivo conditions, enabling precise studies on radiation injury dynamics and therapeutic interventions. The overall objectives of this proposal are to (i) automate perfusion bioreactor and establish parameters to enable immune capability of the skin perfusion system, (ii) determine the utilization of human skin perfusion system to study mechanism(s) of radiation injury and (iii) as a proof-of-principle demonstrate the utilization of human skin perfusion model for developing radiation countermeasure using metformin lotion as an example. Our central hypothesis is that an automated full- thickness human skin perfusion model will provide a robust ex vivo platform for studying radiation-induced skin injuries and developing countermeasures. This hypothesis will be tested through the following three specific aims: 1) Automate perfusion bioreactor and optamize immune capabilities of human skin perfusion model – we will develop a fully automated perfusion bioreactor in collaboration with Daedalus Inc. to improve reproducibility, operational efficiency, and real-time monitoring. Additionally, integrate immune components to enhance the physiological relevance of the model; 2) elucidate the mechanism(s) of radiation induced injuries in human skin - investigate molecular and cellular responses to radiation exposure across diverse human donor samples (sex, age, race) using histological, genomic, and proteomic analyses; and 3) evaluate the use of metformin lotion as a mitigator of radiation induced skin injury – proof -of-principle for employing human skin perfusion model for therapeutics development - as a proof-of-concept, assess metformin lotion efficacy in mitigating radiation- induced skin injuries using the developed perfusion model, demonstrating its potential for therapeutic screening. This project is highly innovative as it will deliver the first-of-its-kind, fully automated, immune-competent human skin perfusion platform, transforming the study of radiation-induced injuries and enabling broader applications for skin-related research. The proposed research is highly significant because it will: i) Provide strong scientific justification for using human skin-based models in radiation injury research; ii) Reduce reliance on animal models, aligning with ethical and regulatory priorities; and iii) accelerate the development and FDA approval process for radiation countermeasures and other skin therapeutics. By successfully achieving these aims, this project will establish a new paradigm in preclinical skin research, bridging the gap between ex vivo modeling and clinical translation to improve radiation injury management and therapeutic development.

NIH Research Projects · FY 2026 · 2026-04

Project Summary Multiple sclerosis (MS) is an autoimmune condition characterized by a relapsing-remitting or progressive course of demyelination and neuron injury, leading to irreversible neurologic disability. While modern therapies reduce MS relapses, there are no available treatments that markedly slow progression or improve remyelination in MS. Recent studies suggest that pathologic immune–glia interactions play a key role in progressive MS. Chronic inflammation transforms astrocytes into reactive astrocytes, which can be toxic to neurons, and impairs remyelination mediated by oligodendrocyte precursor cells (OPCs). The mechanisms that regulate pathologic immune-glial responses in MS are incompletely understood. Interleukin (IL)-33 is a regulatory cytokine and alarmin that promotes protective immune activation, wound healing, and growth factor production. Interestingly, IL-33 is expressed in the healthy CNS and elevated in people with MS (PwMS). Increased IL-33 correlates with decreased lesion burden in PwMS, and IL-33 improves recovery in the mouse model experimental autoimmune encephalitis (EAE), but the underlying beneficial mechanism(s) of IL-33 in neuroinflammation are not fully understood. In other systems, IL-33 is known to potently activate regulatory T cells (Tregs), triggering downstream events including production of the growth factor amphiregulin. In this proposal, I seek to establish IL-33–induced amphiregulin as a therapeutic target to halt progression and promote remyelination in MS. In Aim 1 I will investigate IL-33 mediated Treg activation in EAE. I hypothesize that IL-33 activates Tregs to produce amphiregulin, improving recovery and reducing pathologic reactive astrocytes. I will use transgenic animals where Tregs cannot detect IL-33 and cutting-edge single cell RNA sequencing techniques to characterize the impact on glia cell phenotypes. Aim 2 will test the potential for amphiregulin to enhance remyelination. I will expand on preliminary data showing that recombinant amphiregulin enhances OPC differentiation in vitro. To study myelination in vivo, I will employ an MS-relevant mouse model of demyelination, adoptive transfer cuprizone, using transgenic mice lacking amphiregulin. Finally, IL-33 signaling is blocked by an endogenous decoy receptor called sST2, and my preliminary data showed increased sST2 in the plasma of progressive MS patients. In Aim 3 I will test a novel small molecule that blocks sST2 in EAE. This compound releases the inhibition on IL-33, which I predict will have a beneficial effect through Treg activation and decreased pathologic glia. I will characterize the CNS penetration, pharmacokinetics, and disease impact of this molecule. Throughout this proposal, I strive to employ disease relevant models and readouts that will be instructive for future clinical translation. Completing these aims will advance our knowledge of a promising pathway— IL-33 signaling—to target disease mechanisms in MS that are not addressed by available therapies. Additionally, the proposed training plan, mentorship team, and research aims seamlessly complement one another to facilitate my transition to independence.

NIH Research Projects · FY 2026 · 2026-04

Project Summary Multiple Sclerosis (MS), a chronic autoimmune inflammatory disease of the Central Nervous System, affects approximately 2.3 million individuals globally, with nearly 1 million cases in the United States. It is more prevalent in females and typically manifests before the age of 30. Certain T helper 17 (Th17) cell populations are responsible for neuroinflammation and disease progression. However, patients with hyper-IgE syndrome and HIV infection are susceptible to infections due to the diminished Th17 cell count. Developing strategies to preserve Th17 cells while simultaneously blocking those that cause neuroinflammation and MS remains paramount. Transcriptional analyses have unveiled regulators of Th17 pathogenicity that facilitate immunomodulatory programs and promote pro-inflammatory programs. Nevertheless, mRNA measurements may not accurately correlate with protein abundance, and post-translational modifications such as phosphorylation play a crucial role in engaging signaling networks that drive autoimmunity, which are inadequately captured by mRNA-based approaches. To address this, we will develop a comprehensive suite of mass spectrometry proteomics tools to characterize protein abundance and phosphorylation in pathogenic Th17 cells at the systems level in the experimental autoimmune encephalomyelitis (EAE) model of MS. By comprehending the distinctive signaling pathways that distinguish pathogenic Th17 cells from other CD4 subtypes, we can identify potential targets for alleviating Th17-mediated autoimmune diseases while preserving non-pathogenic populations. Our preliminary proteomics and phosphoproteomics data has already identified novel drivers of pathogenic Th17 activity in the EAE model, which will be validated through small molecule inhibition and genetic approaches. In pharmacological studies, we will identify combinations of small molecule inhibitors that target multiple stages of pathogenic Th17s, offering superior protection against EAE disease compared to monotherapies. We will pursue mechanistic studies to elucidate the synergistic effects of targeting multiple pathways simultaneously in reducing EAE disease. The successful completion of this proposal will advance our fundamental understanding of autoimmune mechanisms mediated by Th17 cells and provide preclinical targets that could be utilized for future translational studies as therapeutic interventions for MS or other Th17-driven diseases.

NIH Research Projects · FY 2026 · 2026-04

ABSTRACT Colorectal Cancer (CRC) is the fourth most common cancer and second leading cause of cancer death in the U.S., with a troubling increasing incidence rate in younger adults. CRC primarily develops from precancerous adenomatous or serrated polyps. Randomized trials have demonstrated that removing these precursor lesions via colonoscopy can prevent subsequent incident CRC. About 7-10% of individuals undergoing colonoscopy have advanced adenomas (AA), defined by size and histologic characteristics. AAs have a higher risk of evolving to CRC and patients with AA have a 3-fold higher rate of subsequently developing CRC compared to those with normal colonoscopy or non-advanced polyps. About 15 million colonoscopy exams/year are performed in U.S., and 25% of them are done for surveillance, or follow up for a history of adenomas. All individuals with AA are recommended to return in 3 years for repeat examination. This recommendation classifies all AA as having similar risk, but the classification of AA is based on endoscopic assessment of polyp size, or villous or dysplasia characteristics on pathologic review, criteria which are subject to interobserver variability. Detailed molecular correlates to define risk are not employed. Robust biomarkers and reliable computational methods for predicting adenoma recurrence in patients with AA based on accurately defined risk could improve patient outcome by personalizing timing of surveillance exams and potentially reducing subsequent CRC incidence, while also optimizing resource allocation and reducing costs. These biomarkers also have the potential to help identify new and effective prevention treatment. We have developed two novel approaches to evaluate colorectal lesions:1. evaluation of aberrant alterations in chromatin structure at nanoscale sensitivity; and 2. spatial analysis of the molecular and cellular diversity of AA microenvironments. We aim to apply these methods to a set of AA tissues from 240 patients whose longitudinal outcome for adenoma recurrence has been tracked Aberrant chromatin remodeling has been shown to be causally linked to neoplastic development. We hypothesize that capturing this remodeling in the index AA has the potential to serve as a sensitive and specific marker of adenoma recurrence risk in individuals post polypectomy. Additionally, the colorectal microenvironment, through its cell-specific molecular mechanisms of interaction is an active sensor of neoplastic transformation. We hypothesize that a microenvironment pre-cancer atlas of AA that leverages the degrees-of-freedom of these interactions can be utilized for developing functional biomarkers that correlate with adenoma recurrence and CRC risk and identify the likely mechanism driving the risk in individual patients who underwent AA polypectomy In Aim 1 we will develop aberrant chromatin organization as a biomarker for predicting adenoma recurrence in patients with AA. In Aim 2 we will generate the molecular and cellular pre-cancer spatial atlas of AA microenvironment at single cell resolution. Together, this multi-scale – chromatin and microenvironment – interrogation offers a promising opportunity to characterize biological determinants of AA risk and to prevent its recurrence and CRC incidence.

NIH Research Projects · FY 2026 · 2026-04

ABSTRACT: One of the most fundamental limitations in medicine is reliance on clinical phenotype as the primary way to diagnose and classify disease. For inflammatory diseases, clinical phenotype alone is not sufficient to describe underlying disease pathogenesis. For example, cytokines like interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-17A can all drive arthritis, but clinical features such as arthritis or rash may not reflect the causal cytokine. As a result, there are no precision biomarkers to guide targeted cytokine therapy: phenotypic classification systems do not stratify patients by causative mechanisms that guide therapy. The lack of precision medicine strategies is one of the most significant unmet needs in the field, leading to poor outcomes for over 40 million patients with inflammatory diseases. This includes >1 million patients with a phenotype-driven diagnosis of “rheumatoid arthritis”: 40-50% of whom fail first biologic therapy, and 10% with disease refractory to multiple consecutive targeted therapies. We propose addressing this problem by combining expertise in immunogenetics and translational immunology (Dr. Schwartz); systems immunology and machine learning (Dr. Das); and statistical genetics (Dr. Wang).We will leverage another key finding: shared molecular mechanisms like IL-6 signaling can drive different phenotypically defined diseases like arthritis, vasculitis, and scleroderma. Using machine-learning approaches, we will build a phenotype-blind molecular taxonomy for immune diseases. Drawing from the success of phenotype-blind tumor genotyping in cancer therapy, we will adapt machine learning approaches developed in the Das lab to build novel tools (SIDER, Significant Interaction Directional Effect Representation; MAGen Multiscale network Approach for Genetic architecture). We will use SIDER and MAGen to construct an “omnigenic model”, connecting a small number of disease-causal core genes to many peripheral genes through complex biological networks. We will ground our approach in human biology using Dr. Schwartz’s expertise to build on >500 core genes that cause monogenic inborn errors of immunity (IEI). We will use interpretable machine-learning and network approaches developed by Dr. Das to develop molecular modules that will be applied to large population databases leveraging Dr. Wang’s expertise We will investigate two aims: (1) understanding monogenic/oligogenic disease architecture and (2) developing novel molecular endotypes for common diseases. This approach will demonstrate a molecular taxonomy is a relevant and actionable approach that will transform the identification, study, and treatment of immune diseases and beyond.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY T cell mitochondrial metabolism is essential for immune cell functions, including immune surveillance and pathogen neutralization. Conversely, impaired mitochondrial function disrupts T cell activity, often resulting in autoimmune diseases, immunodeficiencies, or malignancies. Yet, our understanding of T cell metabolism and its roles in immunologic diseases is poorly understood. Recent work has provided key clues: 1) Disturbances to immune cell metabolic function often result in disease at two opposing ends of the spectrum: cancer and autoimmunity. 2) Rescue of diseased T cell metabolism restores endogenous T cell function, mitigating both cancer and autoimmunity. Moreover, mitochondrial structure and function are intrinsically linked where respiratory complexes do not function in isolation within mitochondria. Rather, these complexes are organized into higher-order assemblies that are concentrated within the cristae and arranged into multi-complex associations of predefined composition, termed supercomplexes. Different disease states not only disrupt the structures of individual complexes but may also alter supercomplex organization to produce symptomatic mitochondrial bioenergetic dysfunction. However, supercomplex formation has never been directly visualized or measured in T cells. I have developed new approaches to directly visualize 3-dimensional mitochondrial structures in healthy and diseased states in patient and animal cells via in situ cryo-electron tomography (cryo- ET). Using biochemical and cryo-ET studies, my goal is to identify the underlying metabolic changes in T cell mitochondrial structure and function during healthy and disease states. I hypothesize: 1) T cell stimulation results in direct structural changes to the respiratory complexes and their higher order organization into supercomplexes; 2) distinct changes in T cell respiratory complex structures contribute to pathologic metabolic dysfunction. To test this, I will identify T cell physiologic mitochondrial ultrastructure and supercomplex stoichiometry (Aim 1) and identify the contributions of T cell mitochondrial ultrastructure and supercomplex stoichiometry within a melanoma tumor microenvironment (Aim 2). Overall, my work will detail how respiratory complexes and their supercomplex organization are regulated in T cells in health and malignancy for the first time. This work may provide a better understanding of T cell pathology, resulting in more effective structure-guided therapeutic interventions. These studies also provide me with training in immunology, biochemistry, and structural biology critical for my development as a physician-scientist.

NIH Research Projects · FY 2026 · 2026-04

Abstract Despite advancements in HIV prevention and treatment and the expansion of services, HIV prevalence among men who have sex with men (MSM) in Vietnam has risen dramatically, from 6.6% in 2015 to 12.5% in 2023. Persistent stigmatization of MSM as "social evils," exacerbated by government campaigns associating MSM with the spread of HIV during the early period of HIV epidemic in Vietnam and reinforced by collectivist cultural norms against homosexuality, has exacerbated intersectional stigmas among MSM in Vietnam. This has led to discrimination, fear of disclosure, and avoidance of healthcare services, severely limiting MSM's access to HIV prevention and care services. To address these challenges, this study will culturally adapt a patient-provider stigma-reduction intervention-Finding Respect and Ending Stigma around HIV (FRESH) intervention- into DONGHANH ( meaning “companionship, understanding, and mutual support" in Vietnamese), for implementation among Vietnamese MSM (the patients) and healthcare providers (HCP- the providers). FRESH is a workshop-based intervention that has been successfully used to reduce stigma among healthcare workers and marginalized populations, including MSM, globally. Toward the end of the DONGHANH intervention, participants are expected to work together to develop stigma reduction strategies that they can take back to their respective communities, thus increasing the impact of the intervention. The intervention will also involve the creation of an e-module (a website), allowing participants to determine its delivery and engage in ongoing learning at their convenience. The study will be conducted in three phases: Phase I: using the ADAPT-ITT framework, we will adapt FRESH intervention based on findings from in-depth interviews and focus group discussions to create the culturally tailored DONGHANH intervention. Phase II: We will use a randomized wait-list control trial design to pilot test DONGHANH intervention among 180 participants (MSM=120; HCP=60). Ninety participants (MSM=60, HCPs=30) will participate in the intervention, while the remaining 90 participants (MSM=60, HCPs=30) will be assigned to the 3-month wait-list control. We will assess the intervention’s feasibility, acceptability, and fidelity as well as assess the preliminary efficacy of the intervention on reducing experiences of intersectional stigma and discrimination, increasing HIV testing, PrEP uptake and PrEP/ART adherence. Phase III: We will evaluate assess facilitators and barriers to implementation through interviews with MSM, HCP and project intervention staff, using the Consolidated Framework for Implementation Research to identify contextual factors of the intervention for future scale-up. If the DONGHANH intervention is successful, it can provide a scalable model for reducing intersectional stigmas and improve HIV prevention and care among MSM in Vietnam and other low-and middle- income countries.

NIH Research Projects · FY 2026 · 2026-04

Abstract Despite the high prevalence of major depressive disorder (MDD) and its projected rise as the leading cause of global disease burden by 2030, treatment efficacy remains suboptimal. First-line antidepressants have modest efficacy (~50%), and high placebo response rates (~40%) contribute to the failure of antidepressant trials and hinder new drug development. While research underscores the role of antidepressant expectancies in modulating mood across various brain regions, there is a critical need to elucidate how expectancy-driven neural dynamics interact with downstream mood regulation processes to induce sustained mood improvement. Our recent work provides the first computational account of antidepressant placebo effects, where reinforcement learning (RL) model-predicted expectancies—encoded in the salience network (SN)—trigger mood changes perceived as reward signals, which reinforce antidepressant expectancies through an expectancy-mood loop. Furthermore, we and others have demonstrated that enhanced functional connectivity (FC) between the SN and default mode network (DMN) during expectancy processing and at rest predicts long-term antidepressant placebo effects. This evidence suggests that antidepressant expectancies, originating from contextual treatment cues, are represented in the SN and influence mood regulation through top-down connections with the DMN. To test this hypothesis, this study will investigate the causal roles of the SN, DMN, and SN-DMN FC in antidepressant placebo effects using Theta Burst Stimulation (TBS). In a 2x3 factorial design, 200 patients with MDD will be randomized to three counter-balanced TBS conditions (intermittent, continuous, and sham, within-subject) targeting either the SN or DMN (between-subject). These acute experimental manipulations will modulate trial-by-trial expectancy and mood ratings and the neural encoding of model-based expectancies and mood reward signals during the “antidepressant placebo fMRI task”, which manipulates placebo-associated expectancies using visually cued fast-acting antidepressant infusions and sham visual neurofeedback. Led by experts in placebo effects, reinforcement learning, depression, and neuromodulation, this study combines a robust theoretical framework, state-of-the-art neuroimaging, precision functional mapping for personalized TBS targeting, and accelerated TBS, ensuring scientific rigor. The insights gained from this study will deepen our understanding of the neural mechanisms behind placebo effects, enhancing clinical trial design, advancing neuroimaging predictors of treatment response, and accelerating the development of expectancy-based interventions for MDD.

NIH Research Projects · FY 2026 · 2026-04

Abstract Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by loss of tolerance to self- nucleic acids and associated protein antigens (Ags), driven by aberrant activation of innate and adaptive immunity. B cells, T cells and myeloid cells infiltrate tissues of lupus patients and mice, causing damage to multiple organ systems including the lung, liver and kidneys. B cells are essential for both the initiation and propagation of lupus disease in murine models. Human SLE is demonstrably B cell-driven, as reflected by the efficacy of FDA-approved and investigational therapeutic drugs including belimumab (anti-BAFF), B cell depleting antibodies, and anti-CD19 CAR T cells. However, these therapeutic approaches target all B cells, leaving patients partially immunocompromised. An approach targeting only the pathogenic subset(s) of B cells is highly desirable. A deeper understanding of which B cell subsets are pathogenic and how they mediate disease is necessary for designing such targeted therapeutic approaches. Over the past several years, a subset of B cells termed "age-associated B cells" (ABCs) was identified as a putative pathogenic autoreactive driver of lupus disease in mouse models and in human patients. These cells are defined by the expression of β2-integrins CD11c (Itgax) and/or CD11b (Itgam) and transcription factors Tbx21 and Zeb2, and are generated through an extrafollicular (EF) B cell response driven by TLR7/9 signals in the presence of IFNγ and IL-21. In addition to being associated with multiple autoimmune diseases, they are formed during infection and may play both protective and pathogenic roles in those settings. The identities, functions and heterogeneity of ABC-like cells and other B cell subsets that infiltrate organs in SLE are not well-defined, yet their evolving relevance in many disease contexts renders defining these an important task. We recently showed in the MRL/lpr lupus model that deletion of CD11c+ B cells reduced disease, including interstitial nephritis, suggesting that these cells promote activation and expansion of autoreactive T cells. What remains unclear is whether ABCs are the sole non-redundant drivers of disease or whether instead they play partial or even dispensable roles, an issue that will be directly addressed by this proposal. Intriguingly, new preliminary data suggests ABCs are themselves present in the kidney and other lupus target tissues of diseased mice, and a recent paper suggests the same is true in human lupus. We hypothesize that ABCs are important drivers of lupus pathogenesis, and that they function in part by presenting self-Ags to autoreactive T cells in the spleen and locally in target organs. Here we propose to define and investigate the pathogenic functions of tissue-infiltrating B cells, including ABCs and other memory B cells, in murine lupus models via two specific aims: 1) Characterize tissue-infiltrating B cells and evaluate their roles in lupus pathogenesis by identifying organ-specific and cross- tissue B cell clonotypes and gene regulatory networks; 2) Genetically test the roles of ABCs in lupus pathogenesis using novel mouse models to inducibly deplete ABCs or specific genes within them.