Northwestern University

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Esophageal motility disorders and gastroesophageal reflux (GERD) are associated with major symptoms (heartburn, chest pain, regurgitation), malnutrition, and aspiration. Nearly 20–30% of the adult population experiences weekly GERD symptoms. Most of the 1 million outpatient visits per year for dysphagia (difficulty swallowing) in the US will require endoscopy which is both invasive and expensive. Additionally, many patients will require further esophageal testing with manometry, reflux testing, and esophagram after endoscopy to identify the underlying cause and identify patients appropriate for surgery. The total expenditure for the management of esophageal disease is estimated to be over $12B/yr and thus, the cost to society is substantial. Esophageal diseases occur when the delicate interplay of neuromyogenic activity, luminal geometry, and esophageal wall mechanics goes awry. Clinical evaluation and management of these disorders utilize endoscopy, esophagram, and high-resolution impedance manometry to characterize anatomy, bolus transit, and neuromuscular function. Although these approaches can identify pathologic patterns for disease classification, they are limited in their ability to guide treatment decisions and determine prognosis due to an inability to assess the effect of physiomechanical factors driving bolus transit. Realizing these limitations, through the past decade, we have integrated physiologic data into physics-based models to simulate esophageal peristalsis and bolus transit. Following multiple iterations and validation, we have generated our current version of the virtual esophagus (vEsophagus). The vEsophagus simulates a virtual twin and effectively resolves the deformation of the esophagus, bolus location, pressure/velocity, stresses/strain in the wall, and neural activation versus time. The scientific premise of this proposal is that we can utilize the vEsophagus to define the physiomechanical perturbations driving poor outcome in patients with achalasia (Aim 1) and GERD (Aim 2) and that we can define an optimal treatment approach using vEsophagus simulations of the virtual twin. We will study patients undergoing directed treatment (POEM, antireflux surgery) in achalasia and GERD patients to determine whether the vEsophagus can define the physiomechanical predictors of disease (Aims 1a & 2a) and can be used to predict outcome after POEM (Aim 1b) and antireflux surgery (Aim 2b). This work will be paradigm shifting due to an entirely new approach to esophageal diseases that focuses on individual patient models (personalized) which can inform treatment selection and prognosis. Tailoring POEM to prevent poor outcomes and complications of blown-out-myotomy and GERD is clinically important, and currently there is no model to guide fundoplication approach to prevent complications of abnormal bolus transit. Our preliminary data provides the rigor to support that we can predict outcome using vEsophagus and we will test this clinically. Eventually, the vEsophagus could replace or reduce clinical trials in humans. It can help predict surgery outcome and could be used to test various motility approaches including pharmacological interventions.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Uncertainty is a part of daily life, but those with difficulty tolerating uncertainty are at risk for anxiety-related and other internalizing disorders. A key contributor to difficulty with uncertainty is an elevated aversion to choices that have ambiguous outcomes (aka ambiguity aversion [AA]). Studies, however, have not tested (a) AA as a transdiagnostic dimension of internalizing psychopathology, and (b) the time course of the brain’s responses to ambiguous choices. This study thus uses computational modeling, electrophysiology, and a novel experimental manipulation to test whether AA is a behavioral and neural mechanism of internalizing psychopathology in a transdiagnostic sample of 140 adult participants with moderate to high intolerance of uncertainty. We will test whether computational parameters of behavioral AA (Aim 1) and event-related potentials [ERPs, neurophysiological markers with millisecond resolution) to ambiguity (Aim 2) exhibit convergent validity (associations with anxiety and depression dimensions, intolerance of uncertainty) and discriminant validity (lack of associations with sensation seeking). Aim 3 will further test AA as a mechanism of internalizing disorders by testing whether an experimental manipulation (i.e., a brief behavioral intervention) “moves” AA. This experimental approach not only provides a robust test of AA as a mechanism within internalizing psychopathology, but also informs future clinical trials that could, for example, use an uncertainty-focused intervention as an adjunct treatment to existing interventions. These aims are novel in two additional ways. First, collecting ERPs related to AA is novel and cost-effective (relative to prior neuroscience studies which have used fMRI), and will provide insight about the time course about the brain’s responses to ambiguity (e.g., whether AA is driven by early [i.e.,~100ms], or later [after 300ms] reactions to ambiguity). Second, while past studies have examined AA in only single disorders, this project will examine associations with transdiagnostic dimensions that theoretically relate to AA (convergent validity) and dimensions that should theoretically not relate to AA (discriminant validity), a rigorous approach that will ultimately have larger clinical impacts. In sum, this project has the potential to identify a transdiagnostic intervention target using an easily administered behavioral and brain assessment of AA, a goal that is directly in line with NIMH’s Strategic Objectives. This K23 will also provide the applicant with the training and mentored research experience necessary for his long-term goal of studying disrupted brain and computationally-derived behavioral mechanisms of internalizing disorders. Training will focus on 1) computational modeling of decision-making behavior, 2) experimental target engagement approaches to interventions, 3) advanced electrophysiological methods and dimensional, transdiagnostic “RDoC” models of psychopathology, and 4) development of grantsmanship, lab management and mentorship skills. This training will be conducted at Northwestern University and facilitated by a team of experts in computational, psychophysiological, and therapeutic approaches, all studying mechanisms of change in internalizing disorders.

NIH Research Projects · FY 2026 · 2025-01

Reducing access to firearms, a modifiable risk factor present in 40% of homes in the United States, is a population-level suicide prevention strategy. S.A.F.E. Firearm is a universal evidence-based suicide prevention intervention primed for implementation nationally. In the largest hybrid type III effectiveness-implementation trial of its kind, funded by NIMH (“ASPIRE” trial), we tested how to implement S.A.F.E. Firearm across 45,000 well-child visits in 30 pediatric primary care clinics at Henry Ford Health (HFH) in Michigan and Kaiser Permanente Colorado (KPCO). We found that training, an electronic health record (EHR)-based clinician decision support (“nudge”), and facilitation (i.e., implementation support to clinics) resulted in robust practice change from 3% reach (baseline) to 54% reach (active implementation). While the ASPIRE trial focused on delivery of the intervention to parents of young people, we have the rare opportunity to expand S.A.F.E. Firearm delivery for all adults via adult primary care and women’s health clinics, offering a new context and broadened population. This holds promise given that (a) many of these patients will have children in their homes (e.g., parents, grandparents) and (b) adults may benefit from S.A.F.E. Firearm themselves because putting time and space between adults and loaded firearms can reduce firearm suicide. The proposed study (“SCALE-ASPIRE”) is motivated by HFH and KPCO constituents’ eagerness to implement this intervention beyond pediatric primary care and will advance the science of implementation by understanding how to “scale-out” successful intervention and implementation efforts. In SCALE-ASPIRE, we will conduct a stepped wedge cluster randomized hybrid type II effectiveness-implementation trial in 48 clinics in adult primary care and women’s health at HFH and KPCO. In Aim 1, in collaboration with key partners including clinicians, leaders, and patients, we will use the ADAPT-ITT approach to adapt S.A.F.E. Firearm for the new clinic context and broadened population. In Aim 2, we will test the effectiveness of S.A.F.E. Firearm and our implementation approach (training, EHR nudge, facilitation) on effectiveness (firearm storage behavior, co-primary; suicide attempts, suicide deaths, and all-cause firearm injury and mortality in adults and young people, secondary) and implementation (reach, or patient-reported receipt of S.A.F.E. Firearm, co-primary; implementation fidelity and cost, secondary) outcomes. In Aim 3, we will use mixed methods to elucidate intervention and implementation mechanisms and to better understand heterogeneity across levels. Specifically, we will conduct interviews to understand constituent perspectives, and explore how patient, clinician, and clinic factors relate to heterogeneity in our co-primary outcomes. Study results will guide the identification of effective secure firearm storage interventions, increase knowledge of effective implementation strategies in large health systems, and advance the literature on scaling-out successful intervention and implementation efforts to new populations and settings, in line with Firearm Injury and Mortality Prevention Research, NOT-OD-23-039.

NIH Research Projects · FY 2026 · 2025-01

U.S. Vietnamese individuals (i.e., those living in the U.S. and identifying as Vietnamese) experience high rates of HPV-related cancers. Although effective strategies exist to prevent HPV-related cancers, uptake of HPV prevention measures remains low among U.S. Vietnamese adolescents. Digital health can potentially be an effective approach to deliver tailored interventions targeting HPV prevention among U.S. Vietnamese, especially given the high rates of internet use in this population. Yet, few digital health interventions on HPV prevention exist for U.S. Vietnamese. More critically, existing digital health research on HPV prevention for Asian Americans and/or U.S. Vietnamese has typically employed the classic “treatment package” approach where different intervention components are bundled into one package and the entire package is tested in a trial. This approach fails to identify either the individual effect of each component or whether the effect of one component is changed by the presence of other components. Understanding distinct impacts of intervention components is critical for ensuring intervention efficiency, affordability, scalability, and effectiveness. This study, HPV Education & Resources for the U.S. Vietnamese Population (HERO), will address this critical gap. Dr. Ha Ngan (Milkie) Vu will leverage the Multiphase Optimization Strategy (MOST) framework to conduct a pilot trial testing four different intervention components (expert video, self-persuasion, narrative storytelling, and motivational interviewing) targeting HPV prevention among U.S. Vietnamese. Additionally, she will apply an implementation science framework to explore factors influencing the future implementation of the intervention. This study represents the first systematic effort to assemble an optimal digital HPV prevention intervention for U.S. Vietnamese. The specific aims are to: (1) Assess the feasibility and acceptability of each of the four digital HERO intervention components in a pilot trial; (2) Investigate effects of each component on HPV prevention outcomes and psychosocial mediators; and (3) Explore factors that may influence the future implementation of HERO. This research is responsive to NCI’s priorities in cancer prevention and digital health. The proposed K01 research activities build on Dr. Vu’s formative research and are supported by robust community and clinic partnerships. They also align with her career development plan, which includes training in (1) clinical trials design, conduct, and evaluation; (2) digital health; (3) implementation science; and (4) professional development. Dr. Vu has assembled a strong team of NIH-funded mentors and advisors with extensive experience in mentoring junior investigators to independence. The K01 will be foundational to Dr. Vu’s career goal of becoming a leading independent researcher who develops, implements, and tests tailored, evidence-based digital health interventions to improve cancer prevention outcomes.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY: There is an urgent need to identify biomarkers that can predict the development of inflammation-associated comorbidities in people with HIV (PWH) on antiretroviral therapy (ART), as well as to develop novel strategies to prevent or treat these comorbidities. Our recent data suggest that glycomic alterations in circulating IgGs are not only candidates for such biomarkers but they may also mechanistically contribute to HIV-associated inflammation by compromising antiviral immuity in a manner that can be normalized In the general population, IgG glycomic alterations drive inflammatory responses during aging (inflammaging). In a recent publication (Giron et al., Nature Communications, 2024), we found that living with ART-suppressed HIV infection is associated with an acceleration in the accumulation of pro-aging IgG glycomic alterations. Specifically, antibodies from PWH on ART showed a significant loss of the anti-inflammatory glycans galactose (agalactosylation) and sialic acid (hypo-sialylation) compared to HIV-negative controls. These alterations were associated with greater inflammation and increased severity of inflammaging-associated comorbidities in PWH on ART. In proof-of-concept studies, we also found that these alterations might precede the development of such comorbidities by years, making them viable candidates for discovering predicitive biomarkers. Beyond their potential as biomarkers, agalactosylation and hypo-sialylation could also constitute causative mechanisms contributing to inflammation in PWH on ART: 1) Agalactosylation: Glycans on IgGs regulate the binding of IgGs to their Fc receptors, defining the ability of IgGs to elicit anti-viral innate immune functions. We examined whether the lack of galactose on IgGs compromised their anti-HIV Fc-mediated immune functions; we glyco-engineered the HIV antibody 10-1074 to produce agalactosylated and highly-galactosylated glycoforms. Agalactosylated glycoforms exhibited significantly lower anti-HIV immune function than highly-galactosylated glycoforms. We propose that this compromised anti-HIV immune function, caused by IgG agalactosylation, impairs the immune system's ability to control virally-infected cells, leading to greater inflammation. Consistent with this hypothesis, we found IgG agalactosylation during ART correlated with elevated levels of HIV DNA during ART and faster HIV viral rebound after stopping ART. 2) Hypo-sialylation: Sialic acid can initiate an anti- inflammatory response by binding to inhibitory molecules on myeloid cells. Loss of sialic acid hinders this mechanism. Our data support this mechanism by showing that treating humanized mice (no ART) with sialidase inhibitors (to prevent hypo-sialylation and preserve sialic acid) attenuated viremic HIV-associated inflammation. These data support our overarching hypothesis that agalactosylation and hypo-sialylation: 1) can serve as predictive biomarkers for inflammaging-associated comorbidities in PWH on ART (Aim 1); 2) are mechanistically linked to HIV persistence and inflammation by compromising anti-viral Fc-mediated innate immune functions (Aim 2a, 2b); and 3) can be normalized to enhance immunity and inhibit inflammaging (Aim 2b, Aim 3).

NIH Research Projects · FY 2025 · 2025-01

Summary The Lymphatic Forum 2025 (LF2025) will be the 5th iteration of a planned conference series that brings together researchers from around the world who study lymphatics in health and disease. The lymphatic system is a relatively understudied, yet vital component of the cardiovascular system that couples the tissue parenchyma to the venous blood via lymph flow through a complex network of lymphatic vessels. It transports fluid, macromolecules, lipids, tissue catabolic products, and immune cells/agents as lymph. Thus, the lymphatic system provides critical links for the transport of fluid, macromolecules, cells, and other elements from the interstitial space of all organs to and through numerous lymph nodes to the venous blood. In completing this transport, it regulates body fluid volume, macromolecular homeostasis, immune function, inflammation, and lipid absorption/metabolism. It is a specialized vascular system with a complex network of low-pressure vessels with unique mechanisms to form and transport lymph. The lymphatic system serves as both a complex network of lymph conduits and as a series of unique intrinsic lymph pumps that generate and regulate the lymph flow that drives all of its functions. Because of its unique, complex structural and functional characteristics, it has been difficult to examine and quantify its structure and function in the body. However, a better understanding of its structure and function is important to health and disease. Other international/national lymphatic biology conferences, including the previous Lymphatic Forum 2017, 2019, 2021 (Virtual), and 2023 focused predominantly on embryologic development, lymphangiogenesis, lymphatic endothelial cell function, and relationships to cancer and immunity. The conference plan outlined in this application is unique in that it is the first that has a significant focus on disease-oriented sessions on the lymphatic system, including lymphatic disease detection, tumor lymphatics and lymph node metastasis, therapeutic innovations, and clinical trials. Presentations and discussions will focus on the molecular regulation of the lymphatic system in health and disease, exploring new insight into its contribution to pathological processes. Additionally, the conference will also include a plenary session on unsolved issues and gaps in lymphatic disease with a panel discussion. The lymphatic system is an intrinsic, understudied part of the cardiovascular and immune systems that influences health and disease. The goal of LF2025 is thus highly relevant to the missions of the NHLBI which has accepted primary assignment on this application, and also of the NIDDK, NIAID, and NCI, which will be asked for secondary support. Additionally, the conference is specifically designed such that trainees and young investigators have significant roles.

NIH Research Projects · FY 2026 · 2025-01

Summary Pragmatic (i.e., social) language impairments are a defining feature of autism spectrum disorder (ASD), which can impose significant burden on individuals throughout the lifespan. Strong evidence also suggests that this clinical domain is influenced by genetic markers associated with ASD in clinically unaffected relatives, constituting a principal component of the broad autism phenotype (BAP). Although sex differences in the development of pragmatic language have been repeatedly demonstrated in the general population, sex-specific differences in pragmatics in ASD and the BAP are woefully understudied, despite mounting evidence of a distinct clinical phenotype among autistic females, and the strong impact of pragmatic skills on clinical outcomes and well-being. Most studies of pragmatics in ASD have included only males or included so few females that they were insufficiently powered to address the potential of sex-specific patterns of pragmatic skills in conversation, narrative, or other crucial social-communication contexts where pragmatic skills play an essential role in social functioning (including vocational success) across the lifespan. In this project, we will apply an armamentarium of deep phenotyping methods to dissect the key features and contributors to the hypothesized sex-specific pragmatic language profiles of ASD and the BAP that extend beyond traditional, categorically-defined diagnostic boundaries, using a family-study design and including a representative ASD cohort enriched for females, ASD parents, and respective control groups. Aim 1 will characterize the pragmatic language profile of females with ASD and related, potentially sex-specific neurocognitive mechanistic skills and psychosocial outcomes. Aim 2 will conduct parallel questions in the BAP. Finally, Aim 3 will estimate the incidence of sex-specific pragmatic profiles of ASD and BAP in the general population via crowdsourced pragmatic phenotyping. Our preliminary studies demonstrate strong evidence for complex sex-specific patterns of pragmatic abilities in ASD that may be comprehensively and sensitively measured with the complement of gold-standard hand-coding methods and advanced fine-grained computational- and machine-learning-based approaches proposed in this project. Together, the rich and extensive data produced will contribute to our understanding of the fine-grained skills underlying the sex-based heterogeneity in ASD, and the mechanistic, potentially sex-specific origins of pragmatics as an important clinical domain. Our findings will move us closer to achieving long-term translational goals to advance understanding of the sex-specific differences in pragmatic language impairment in ASD in order to improve the prediction of clinical outcomes and help guide future interventions and treatment in ASD.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY: The blood-retina barrier is a key anatomical characteristic of the retina, which is disrupted in diabetic retinopathy (DR), retinal vein occlusion, and posterior uveitis. Retinal perivascular macrophages are components of the blood-retina barrier, yet their function is unknown. Brain perivascular macrophages are long-lived tissue resident macrophages that reside on arterioles and venules, which regulate blood-brain barrier hyperpermeability. Our preliminary data demonstrate that retinal perivascular macrophages are the major antigen presenting cells in the retina and reside on post-capillary venules. Furthermore, retinal perivascular macrophages express a pro- chemotactic transcriptome for monocytes, neutrophils, and lymphocytes. Additionally, we developed an effective tool to target and deplete (Pf4Cre) perivascular macrophages without affecting microglia. Finally, after intravitreal CCL2 injection, perivascular macrophage depletion severely hampers Ly6C+ monocyte infiltration. Based upon these data, we hypothesize that retinal perivascular macrophages are long-lived tissue resident macrophages that orchestrate transendothelial migration of immune cells during DR and uveitis. To test this hypothesis, we formulated the following specific aims: 1) Determine the ontological origin of retinal perivascular macrophages. In this aim, we will use Cx3cr1CreER x Rosa26GFP mice to fate map perivascular macrophages. We will label macrophages at the yolk sac and fetal liver stages to determine perivascular macrophage origin. Additionally, we will perform pulse-chase labeling studies to determine the longevity and bone marrow contribution over 1 year. 2) Determine the role of perivascular macrophages during experimental autoimmune uveitis (EAU). We will use Pf4Cre x Rosa26LSL-DTR mice to deplete perivascular macrophages during EAU. Additionally, we will use Pf4Cre x MHCIIflox mice to conditionally knockout MHCII in perivascular macrophages during EAU. 3) Determine the role of perivacular macrophages during diabetic retinopathy (DR). In this aim, we will use diabetic Pf4Cre :: Csf1rflox/flox mice to chronically deplete perivascular macrophages and perform confocal immunofluorescence of retinal flatmounts to quantitate immune cell infiltration at Month 1 and both pericyte numbers and avascular capillaries at Month 9. Completion of these aims will determine that perivascular macrophages are derived from fetal progenitors without bone marrow contribution, are necessary for EAU initiation, and regulate immune cell infiltration during DR. The blood-retina barrier is a key component in health and blinding diseases like uveitis and DR. These studies will significantly advance our knowledge of perivascular macrophages and their blood-retina barrier functions. Furthermore, they will potentially open a new therapeutic space, targeting perivascular macrophages during inflammation and inflammatory retinal vascular diseases.

NIH Research Projects · FY 2026 · 2025-01

While with the recent breakthrough in immunotherapy, which has transformed into a standard care for therapy in treatment of many advanced cancers, cancer remains as the second leading cause of death. This is in part due to the efficacy of cancer immunotherapy is still limited in treatment of most solid tumors and often associated with immunotherapy-related adverse events with multiple tissues/organs autoimmune inflammation. Clinicians have attempted combinations of cancer immunotherapy with either irradiation or chemotherapies to achieve synergistic efficacy and reduce their toxicities. However, one of the obstacles is that many of the chemotherapeutic drugs, if not all, often have immune toxicities, and their administration largely reduces the immune cell frequency in the circulating blood, which consequently impairs antitumor immunity and diminishes immune checkpoint blockade therapeutic efficacy. Another urgent need in cancer therapy is to improve the therapeutic efficacy to target cancer stem cells, which drive cancer inhiation, metastasis and recurrence. Therefore, it will be ideal to develop an antitumor strategy with both chemo-targeting and immuno-activating therapeutic efficacy and also can target cancer stem cells. Importantly, our recent publications and preliminary studies revealed that USP22 functions as an oncogene in tumor cells and inhibits immune surveillance. Genetic USP22 suppression inhibits cancer cell growth, impair cancer cell stemness and boost antitumor immunity. The current application from Northwestern University (NU) and an industry partner, the startup company ExoMira, with the service based support from a world-leading chemistry company WuXi AppTec aims to develop IND-enabling study of a first-in-class medicine, USP22- specific small molecule inhibitor, for the treatment of triple negative breast cancer.

NIH Research Projects · FY 2026 · 2025-01

SUMMARY Each year in the United States there are 14,000 bladder augmentation enterocystoplasty surgeries to address trauma, urological cancers, severe cases of spina bifida, and interstitial cystitis. Although enterocystoplasty is the standard of care for patients with end-stage pathologic bladder, 33% of patients have complications due to anatomical and physiological differences between bladder and bowel tissue used to augment bladder capacity. Complications include bladder perforations, renal failure, and malignant transformation. There is currently no viable alternative to augmentation enterocystoplasty. Several strategies have been reported to replace enterocystoplasty and regenerate bladder tissue, but these have failed clinically. Reasons for the failure include the common use of phylogenetically dissimilar pre-clinical animal models that do not accurately represent the human bladder or its disease condition, the use of inadequate materials to serve as scaffolds for cells to grow on and regenerate bladder tissue, the use of often diseased autologous bladder cells that have lost the capacity to regenerate functional bladder tissue, and an inability to continuously monitor the tissue regeneration process to identify potential problems at an early stage. The overall goal of this proposal is to accelerate bladder tissue formation and enable wireless, real-time monitoring of bladder function. Toward this goal, we have demonstrated that: 1) we can restore normal bladder function at 6 months through 24 months post-surgery in a clinically relevant baboon bladder augmentation model using a new mechanically compatible biodegradable elastomer scaffold, poly(1,8 octamethylene citrate-co-octanol) (POCO) seeded with autologous baboon bone marrow-derived mesenchymal stromal cells (MSCs) and hematopoietic stem/progenitor cells (CD34+ HSPCs). At 24 months, peripheral nerve regeneration was adequate and functional in the regenerated area; 2) engineering scaffold microtopography with parallel microgrooves can improve anatomical features of the regenerated bladder. Specifically, in a nude rat bladder augmentation model, microgrooved POCO scaffolds seeded with human bone marrow-derived MSCs and CD34+ HSPCs, blood vessel density, muscle to collagen ratio, and urothelium thickness were increased relative to cells seeded on scaffolds with smooth surface; 3) bladder contraction events in rats and baboons can be wirelessly detected in real time via a biointegrated electronic strain gauge; 4) stretchable electronics can be integrated into citrate-based elastomers; and 5) electrically conductive POCO scaffolds enable bladder tissue regeneration in a rat bladder augmentation model without the need for seeded cells, simplifying clinical translation of this approach. The specific aims of this proposal are to: 1) investigate electrically conductive and non-conductive microgrooved POCO scaffolds for accelerated bladder tissue regeneration, and 2) investigate wireless bioelectronic strategies to monitor real time bladder dynamics.

NIH Research Projects · FY 2026 · 2025-01

Glioblastoma (GBM) is an incurable brain tumor with a median overall survival of less than 15 months. The recent success of immunotherapy in extracranial malignancies has translated into trials for GBM. Despite its promise, immunotherapy's clinical efficacy has yet to be shown for GBM. The resistance to immunotherapy in GBM is multifactorial. To uncover the targets and pathways contributing to resistance in GBM, we designed and performed an unbiased, genome-wide CRIPR-Cas9 knockout screen screening in a glioma cell line under a cytotoxic T-cell mediated selection pressure. This screen identified SEC61subunit gamma (SEC61G) as a gene involved in immunotherapeutic resistance in GBM. SEC61G is a component of the SEC61 translocon complex in the endoplasmic reticulum. Our data shows that SEC61G is highly expressed in GBM, with genomic amplification seen in up to 30-40% of GBM patients. Knocking out SEC61G sensitized glioma cells to T cell- mediated therapy in vitro. Proteomic analysis of SEC61G knockout cells revealed an interplay of SEC61G expression with the activation of the MAPK pathway in glioma cells. The proposed pre-clinical investigation aims to understand the role SEC61G in immunotherapeutic resistance in GBM and to evaluate SEC61G as a therapeutic target in combination with immunotherapy. We hypothesize that SEC61G confers resistance to T cell-mediated immunotherapy in GBM and that targeting SEC61G can significantly enhance the response of GBM to immunotherapy. This hypothesis will be tested in three Specific Aims. SA1 will investigate how SEC61G overexpression interconnects with the MAPK pathway and contributes to the T cell-mediated therapeutic resistance in glioma cells. SA2 will determine the contribution of SEC61G to the immunosuppressive tumor microenvironment impeding the responses of T cells in GBM. SA3 will investigate SEC61G treatment regimens in combination with T-cell mediated therapies using murine and human models of GBM. Upon completing these studies, we will understand the mechanisms underlying SEC61G-mediated immunotherapeutic resistance, characterize SEC61G as a therapeutic target, and yield translational results for the treatment of GBM and potentially other solid tumors.

NIH Research Projects · FY 2025 · 2025-01

PROJECT SUMMARY Cytomegalovirus (CMV) remains the most common viral infection in kidney transplant recipients leading to significant morbidity and mortality despite preemptive and prophylactic anti-viral therapies. While anti-rejection immunosuppressive therapy is considered a significant contributing factor of increased incidence of CMV infection following kidney transplant, we show that ischemia/reperfusion injury (IRI) inherent to transplant is a main trigger to induce CMV reactivation following murine transplantation of a latently infected graft, in addition to its prominent role in delayed graft function and allograft rejection. Currently, no FDA-approved therapy is available for IRI. The goal of our research is to identify cellular and molecular targets for the development of tissue/cell-specific strategies to prevent CMV reactivation and improve transplant outcomes. Endoplasmic reticulum (ER) stress and mitochondrial (Mt) dysfunction are two interconnected adverse events incurred in IRI. Inhibition of ER stress regulates Mt function and attenuates kidney injury. Inositol-requiring enzyme-1α (IRE-1α), a main transducer of ER stress, has been implicated in CMV infection. Data from our preliminary studies demonstrate that downregulation of IRE-1α expression is associated with reduced kidney transplant IRI by deletion of its upstream mediator (e.g., TLRs/Myd88). Additionally, we show that inhibition of Mt fragmentation by knocking down dynamin-related protein1 (DRP1), a mitochondrial fission protein in ECs, reduced EC activation and graft injury. Moreover, renal endothelial cells (ECs) are the frontline responders to IRI and also a main cell type of CMV latency and reactivation, representing an appropriate target site. However, it has not been investigated whether modulating ER stress and/or Mt function in graft ECs prevents CMV reactivation. The objective of this novel study is to determine whether targeting IRE-1α and/or DRP1 attenuates IRI, thereby preventing CMV reactivation in D+/R- kidney transplant. We hypothesize that CMV reactivation is initiated by IRI-mediated aberrant IRE-1α dependent ER stress responses and DRP1 mediated Mt dysfunction in donor renal ECs that harbor the latent CMV genome. We will use a preclinical mouse model of kidney transplantation to test our hypotheses. In Aim 1, we will first investigate the role of donor-derived IRE-1α in IRI-mediated CMV reactivation in rejection-free syngeneic transplants, and then in allogeneic transplants to address influence of rejection response and in modulating donor EC transcriptome landscape. Additionally, we will investigate the role of donor derived DRP1 in transplant IRI and CMV reactivation using a similar approach. In Aim 2, we will determine the therapeutic potential of pre-operative tissue/cell-specific targeting of ER and Mt stress pathways by leveraging nanocarrier delivery platforms to mitigate kidney transplant IRI, thereby preventing CMV reactivation. Data from this study will lay the foundation for further studies in an ex-vivo perfusion system of human kidneys towards a clinically translatable pre-treatment strategy for donor organ management to mitigate IRI and reduce the risk of CMV reactivation, improving the current standard of care in solid organ transplantation.

NIH Research Projects · FY 2026 · 2025-01

Modified Project Summary/Abstract Section Growing up in a lower-income family robustly predicts worse mental health in adolescence and early adulthood. How does variability in family income “get under the skin” of the developing child and via what mechanisms does it increase risk for mental illness? Moreover, could supplements to family income at critical developmental periods help to prevent later youth mental illness? To address these questions, we leverage an innovative existing double blind randomized controlled trial of 3-years of substantial ($1,000/month vs. $50/month) income supplements to parents. By experimentally studying the impacts of these income supplements on families and subsequent youth development, we can examine causal pathways from family income to risk for mental illness via family stress and neuroimmune mechanisms in ways never done before. Moreover, by measuring the longer-term impact of 3 years of income supplements to parents on their child’s neuroimmune signaling and risk for mental illness, we can examine the policy implications for child development of unconditional cash transfers to parents and identify how and for whom these supplements help. We will test these basic and translational questions in a sample of 1,200 youth with lower-income parents randomly assigned to receive either a substantial monthly income supplement ($1,000/month, intervention group) or a minimal monthly supplement ($50/month, control group) for 3 years, starting when youth were between age 5 – 14 years old. We will follow up with youth and their parent 1 – 2 and 3 – 4 years after the intervention (youth will be age 10 – 18 at first follow-up) and examine whether income supplements predict better youth mental health during adolescence, as well as whether factors like child age and neighborhood quality modulate intervention effects. Additionally, we explore family stress mechanisms through which the intervention may impact child mental health. Finally, we will measure peripheral inflammation (inflammatory biomarkers and classical monocytes) and use MRI to assess threat, reward, and regulatory neural activity and connectivity among 500 of these youth. Our central hypothesis is that income supplements will decrease family and youth stress and improve parenting, which will improve neuroimmune signaling and decrease risk for psychopathology. Moreover, these effects will remain years after termination of the transfers and be strongest among families who received the intervention earlier in the child’s life. This research will provide timely, relevant public health knowledge that will clarify whether cash transfers to parents have longer-term effects on brain, immune, and mental health, while also advancing the science of the sociocontextual and neuroimmune pathways through which variability in family income impacts risk for mental illness.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Lung transplantation is the only viable therapeutic option to extend and improve the quality of life for many patients with end stage lung disease. However, survival following lung transplantation is the worst among solid organs with only 80% and 50% of patients alive at one and five years, respectively. Primary graft dysfunction (PGD) after lung transplantation, resulting from ischemia reperfusion injury, is the predominant cause of poor lung transplant early outcomes. Chronic lung allograft dysfunction (CLAD) with lung fibrosis is the leading cause of morbidity and late mortality after lung transplant. Epidemiologic data suggest a strong link between PGD and CLAD but the molecular mechanisms underlying this association are unknown. Our laboratory has shown that non-classical monocytes in the donor lung are necessary for neutrophil recruitment to the allograft leading to PGD after lung transplantation. In parallel studies, we showed that activation of the integrated stress response in the alveolar epithelium precludes alveolar epithelial differentiation after injury. Based on these published and preliminary data, we hypothesize that early PGD induces a chronic activation of the integrated stress response in the airway and alveolar epithelium that precludes epithelial differentiation in response to injury, predisposing to a risk of CLAD. We will test this hypothesis in three interrelated specific aims. Aim 1: To determine whether ISR (Integrated Stress Response) mediated activation of ATF4 is necessary for impaired alveolar epithelial repair after lung transplantation. Aim 2: To determine whether ameliorating PGD by targeting donor derived nonclassical monocytes reduces the severity of CLAD. Aim 3: To determine whether activation of the ISR in the airway and alveolar epithelium in lung predisposes patients at an increased risk of early CLAD. We will test our hypotheses in clinically relevant murine models of PGD and CLAD following lung transplantation. We will credential our findings in humans by applying multi-omic technologies to the analysis of samples collected longitudinally after lung transplantation. Our studies are designed to credential targets for therapy that can be administered at the time of transplant to reduce the long term risk of CLAD.

NSF Awards · FY 2025 · 2025-01

Global initiatives aim to significantly reduce carbon dioxide (CO2) emissions by 2030 and achieve net-zero emissions by 2050. Electrocatalytic CO2 reduction (e-CO2RR) has emerged as a promising approach to convert CO2 into valuable chemicals and fuels using renewable electricity. However, current e-CO2RR systems face efficiency and stability limitations that hinder their practical implementation. Copper-based catalysts, while selective towards hydrocarbon production, suffer from performance degradation over extended operation. Leveraging a collaboration between Northwestern University in the US and the Fritz Haber Institute of the Max Planck Society in Germany, this project focuses on developing techniques for studying novel catalysts under realistic operating conditions and integrating advanced synthesis, characterization, and modeling methods to unravel the dynamic changes during reaction. Fundamental understandings gained from this project will provide essential guidance for developing catalysts that can be scaled up for real-world applications and enabling large-scale CO2 utilization for chemical manufacturing and carbon management. Beyond the technical aspects, the investigators are deeply invested in educational and outreach initiatives ranging from elementary school children to post-doctoral researchers. Those efforts will continue under the project, with specific efforts focused on training graduate students in the most advanced microscopy techniques relevant to catalyst design and performance under realistic electrochemical reaction conditions. The scientific underpinning of this project is to elucidate the dynamics occurring at the catalyst-electrolyte interface for intermetallic nanomaterial (iNM) electrocatalysts during CO2 reduction and to correlate these changes with catalyst performance. This project leverages the synergy between four key components: (1) machine learning-guided design and synthesis of iNM catalysts; (2) operando electron and X-ray microscopy techniques for nanoscale visualization of catalyst evolution, including electrochemical cell transmission electron microscopy (EC-TEM) and transmission X-ray microscopy (TXM); (3) development of in situ liquid cells with ultra-thin membrane windows to enhance spatial resolution and chemical sensitivity, aiming to achieve roughly 1-nm resolution in liquid environments; and (4) integration of real-time product analysis, adapting differential electrochemical mass spectrometry (DEMS) concepts for CO2 reduction. By bridging the gap between atomic-scale structural investigations and practical catalyst performance in CO2 electrolyzers, this research aims to accelerate the development of high-performing iNM-based e-CO2RR systems, potentially transforming the approach to carbon dioxide utilization and sustainable energy production. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-01

Project Summary: Ovarian cancer poses a significant burden on patients, society, and healthcare system. Despite the success of cytoreductive treatment and chemotherapy as the frontline treatment, many patients still experience metastases and develop cancer recurrence. Unfortunately, the second-line therapy against recurrent/metastasized ovarian cancer remains ill-defined. Tumor infiltrating lymphocytes (TILs) are rare immune cells that migrate from the bloodstream into the tumor to exert protective antitumor effects. These cells can be isolated from the patient’s solid tumor and have been successfully used in cellular immunotherapy. However, production of TILs requires an accessible tumor and involves invasive surgical procedures to isolate these cells followed by a complicated ex vivo expansion process before they are ready to be used for therapeutic purposes. For ovarian cancer, unfortunately the accessible tumor is usually already removed by cytoreductive treatment and thus unavailable when the recurrent/metastasized tumors are detected. Hence, rapid non-invasive means to isolate rare immune cells from alternative natural reservoirs (i.e., blood circulation) are urgently needed to accelerate the development of effective anti-tumor regimens. Microfluidic platforms are powerful tools for the isolation and non-destructive analysis of rare populations of cells based on inexpensive instrumentation and facile fabrication approaches. Recently, we demonstrated the use of our innovative high-performance microfluidic platform technology for unprecedented enrichment of TILs from tumor tissues as well as other circulating tumor-reactive lymphocytes (cTRLs) from peripheral blood. This technology can isolate rare immune cells with high purity and yields while retaining their therapeutic efficacy after expansion. Since frequency of cTRLs in blood is expected to be very low (< 0.005%), we will further develop our microfluidic systems to enable rapid isolation and expansion of rare ovarian cTRLs directly from the blood of syngeneic animal models and human patients to assess their therapeutic potential. In the proposed project we will engineer microfluidic devices to enable rapid isolation of cTRLs directly from the peripheral blood. Through comprehensive characterization of the isolated cTRLs, we will deploy the design- build-test-learn cycle to identify cell-surface markers that can be targeted to improve the enrichment and potency of cTRLs. The new microfluidic system will permit the culturing of the rare cTRLs for ex vivo expansion. We will elucidate the tumor specificity, reactivity, and therapeutic potency of the expanded cTRLs. Based on syngeneic results, we will also isolate cTRLs from clinical specimens, characterize their molecular signatures, and evaluate their anti-tumor efficacy in patient-derived organoid models. The project deliverables will include a first-in-class microfluidic cell sorting and culturing system for rapid isolation and expansion of cTRLs with the potential to impact the American population by making second-line treatment against ovarian cancer more durable and amenable to patients that do not benefit from conventional front-line treatment.

NIH Research Projects · FY 2026 · 2025-01

Abstract/Summary Viral-based cancer therapies for cancer exert powerful antitumoral effects both through direct infection and lysis of tumor cells and by triggering immune responses against the tumor. Currently FDA approved oncolytic viruses are targeting melanoma and bladder cancer, and there is a growing list of viral agents on the horizon. However, current viral-based therapies have limitations in terms of their safety and efficacy, warranting development of novel agents. It has been suggested that an arenavirus like lymphocytic choriomeningitis virus (LCMV) could be attenuated to render it ideal for viral therapy against cancer. In this proposal, we demonstrate the powerful anti- tumor activity of a highly attenuated LCMV strain, r3LCMV. Engineered from LCMV, this virus is exceptionally immunogenic in mice and humans, and r3LCMV vectors expressing HPV antigens are already being clinically tested as a cancer vaccine. However, our data from multiple murine tumor models, including melanoma models, suggests that r3LCMV is exceptionally effective intratumorally (IT) even without expressing tumor antigens. Our group has been developing novel immunotherapies for melanoma with a focus on mucosal melanoma (MM), a particularly deadly melanoma variant of the mucosal surfaces. MM has one of the best immunocompetent models there is: the spontaneous oral MM tumors that occur naturally in pet (companion) dogs. This project will have 2 aims, each evaluating different aspect of this therapy and exploring different clinical settings: Aim 1: To determine whether neoadjuvant r3LCMV treatment improves disease-free survival (DFS) in dogs with naturally occurring MM. We will launch a placebo controlled veterinary trial to evaluate the effect of IT r3LCMV administration in the neoadjuvant setting for dogs diagnosed with MM. Aim 2: To utilize r3LCMV to improve cellular therapy of MM. TIL generated from canine MM tumors treated with r3LCMV or placebo will be compared for phenotype, ex-vivo function and clinical efficacy for the treatment of metastatic disease.

NIH Research Projects · FY 2026 · 2025-01

ABSTRACT/SUMMARY Uterine leiomyomas (LMs, fibroids) are the most important neoplastic threat to women worldwide. As no long- term non-invasive treatment option exists for LMs, deeper insight into tumor etiology is key to develop more effective medical therapies. Accordingly, this proposal is impactful as it suggests a novel etiological basis for the predominant MED12-mutated LM subtype and offers proof of concept for therapeutic intervention in this specific genetic setting. LMs arise from recurrent and mutually exclusive genetic alterations in a limited number of driver genes. Among these, mutations in the RNA Polymerase II Mediator subunit MED12 (mut-MED12) are by far the most prevalent, accounting for 77.4% of LMs. Recently, we showed that LM driver mutations in MED12 disrupt CDK8/19 kinase activity in Mediator, revealing the first and heretofore only known biochemical defect arising from these pathogenic mutations and further implying a new etiological role for CDK8/19 in LM pathogenesis. Herein, we identify signal transducer and activator of transcription 3 (STAT3), a transcription factor implicated in tumor growth and fibrosis, as a direct MED12-dependent CDK8/19 substrate. Physiologically, we show that CDK8/19, by direct phosphorylation of STAT3 serine 727, is a negative regulator of STAT3 transcriptional activity. However, the role of the STAT3 pathway in LM growth and whether this activity is altered by mut-MED12 remains unexplored. Our preliminary data indicate that STAT3 activation by CDK8/19 disruption is a key mediator of mut-MED12-driven LM growth. (i) Compared with wild-type (wt)-MED12 cells, mut-MED12 primary LM cells or a human myometrial smooth muscle cell line with CRISPR-engineered mut-MED12 responded to STAT3 inhibitors with greater sensitivity and decreased growth. (ii) CDK8/19 disruption by mut-MED12 reduced STAT3 serine 727 phosphorylation, leading to an increase in tyrosine 705 phosphorylation and strikingly enhancing the transcriptional activity and chromatin binding properties of STAT3 that drives expression of genes crucial for LM growth. (iii) STAT3-activating cytokines were specifically upregulated in myometrium adjacent to mut-MED12 LM, activated the JAK/STAT3 pathway, and promoted mut-MED12 LM cell growth. (iv) Progesterone receptor binds to the STAT3 gene promoter and stimulates its expression. We hypothesize that Mediator kinase disruption as a consequence of LM driver mutations in MED12 alters the phosphorylation pattern of STAT3 to favor a hyperactive form, one with enhanced chromatin binding properties leading to transcriptional activation of a unique set of genes responsible for LM growth. We propose that STAT3 inhibition using FDA-approved or in- pipeline agents targeting the JAK/STAT3 pathway will block steroid hormone-dependent LM growth, especially for the mut-MED12 subtype. Using a clinically relevant patient-derived xenograft LM mouse model and a cutting- edge single-nucleus multiomics approach, we will: (1) determine the functional role of the STAT3 pathway in LM growth in vivo; and (2) define the cell populations with unique epigenetic and transcriptomic processes responsible for STAT3-driven growth in mut-MED12 LM and verify the mechanistic findings in clinical samples.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Polycystic ovary syndrome (PCOS) is a common, complex, and heritable endocrinopathy, affecting 6-20% of reproductive age women. PCOS is defined by elevated androgens, irregular or absent periods, and polycystic ovarian morphology and is diagnosed by the presence of two or more of these reproductive phenotypes. There are many interacting physiological pathways that contribute to PCOS pathogenesis, including insulin resistance, ovulatory dysfunction, and gonadotropin signaling dysfunction. Due to the multifactorial nature of this disorder, current clinical interventions are variably effective and do not address the underlying causes of PCOS. Twin and family studies have shown PCOS to be highly heritable. Furthermore, genome-wide association studies (GWAS) have implicated genes involved in insulin resistance, diabetes, ovulation, and gonadotropin signaling. Genetic factors, therefore, are likely contribute to the heterogeneity of PCOS. With the advent of Next Generation Sequencing (NGS) it is now feasible to comprehensively identify genetic variation mapping to a genome, gene, or molecular pathway. Our long-term objective is to identify genetic drivers of PCOS. The gonadotropin hormones, follicle stimulating hormone (FSH) and luteinizing hormone (LH), are part of the neuroendocrine signaling network that direct follicle development and overall ovarian health. Reproductive neuroendocrine dysfunction is a major pathophysiological component of PCOS. Women with PCOS have an imbalance of LH:FSH ratio resulting in hyperandrogenism and oligo/anovulation—two cardinal characteristics of PCOS. Although gonadotropin genes have long been implicated in PCOS pathogenesis, no causal variants have yet been identified. Given the physiological relevance of gonadotropin signaling dysfunction and genetic implications of gonadotropin genes in PCOS, our hypothesis is that genetic variation in gonadotropin genes contributes to PCOS pathogenesis. Therefore, our goal is to elucidate the molecular mechanisms of gonadotropin signaling dysfunction in PCOS. In this study, we will evaluate the genetic burden of potentially pathogenic variants (PPVs) in the gonadotropin signaling pathway in women with PCOS using two independent PCOS cohorts (Aim 1). Furthermore, we will elucidate the molecular mechanisms by which these pathogenic variants cause gonadotropin signaling dysfunction through functional studies (Aim 2). Successful implementation of these aims will reveal the extent to which gonadotropin dysfunction is an underlying cause of PCOS, which members of the gonadotropin signaling pathway are impaired in PCOS, and how pathogenic variants in this pathway impair gonadotropin signaling at the cellular level.

NIH Research Projects · FY 2025 · 2024-12

ABSTRACT The goal of this proposal is to test the hypothesis that nasally exhaled breath contains biomarkers of Alzheimer’s disease (AD). Preliminary data from our lab indicate that: (1) the neural protein Tau is present in nasally exhaled breath at higher concentrations than in orally exhaled breath, and (2) Tau concentration increases linearly with dementia severity in patients with AD. An established body of research indicates that exhaled breath contains a rich variety of biological molecules including proteins, cytokines, RNA, DNA and other biomarkers that originate along the lining of the respiratory tract. With the goal of measuring respiratory biomarkers, the vast majority of exhaled breath studies use orally exhaled breath; by contrast, very little research has focused on nasally exhaled samples. In this proposal, we aim to focus our research on nasally exhaled breath in order to test the hypothesis that the unique anatomy of this exhalation route provides direct access to neural biomarkers of AD. Specifically, human olfactory sensory neurons constitute the only part of the central nervous system that makes direct contact with the external environment. The cilia on their peripheral ends protrude from the olfactory mucosa into the nasal cavity, while the central ends project through holes in the cribriform plate at the base of the skull and synapse directly onto the brain. The human cribriform plate is also a drainage pathway for cerebrospinal fluid. As a result of these unique anatomical circumstances, chemical constituents of the central nervous system are present in the olfactory mucosa inside the nose, likely due to nasally exhaled air flowing turbulently over the olfactory mucosa and volatilizing proteins in this region. In this proposal, we aim to explore the novel idea that these volatilized neural proteins can be collected in nasally exhaled breath and quantitated. To test this notion, we will first examine the hypothesis that biomarkers of neurological disease (Tau, beta-amyloid) can be quantitated noninvasively by collecting nasally and orally exhaled breath from patients currently diagnosed with AD, and comparing levels of biomarker concentration across nasal and oral samples. Second, we will compare levels of biomarkers concentration (nasal, oral) across three groups of participants: patients with AD, age-matched healthy controls, and healthy young adults. Across the proposed experiments, we will also look for correlations between nasally (vs. orally) exhaled proteins and disease severity. If successful, results from the proposed studies could aid in the development of a novel, noninvasive and inexpensive diagnostic tool for ADRD and other neurological diseases that could offer mobility and accessibility for at-home testing. Future research in this direction would not only be clinically valuable but could also leave a broader scientific impact by initiating a step forward in advancing current limits of real-time human brain chemistry measurement.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY/ABSTRACT Cholestatic liver disorders affect over 200,000 Americans and there are few effective and no curative therapies to prevent disease progression. The accumulation of intracellular misfolded and unfolded proteins causes a form of cellular stress termed endoplasmic reticulum (ER) stress. The X-box binding protein 1 (XBP1) pathway is a highly conserved unfolded protein response signaling pathway that acts to reduces endoplasmic reticulum (ER) stress and return cells to normal homeostasis. During cholestasis, elevated intrahepatic bile acid concentrations can cause protein misfolding and ER stress in the liver, and we have previously shown in murine models and human liver specimens from patients with cholestatic liver disease that the inability to properly activate the hepatic XBP1s pathway is associated with enhanced cholestatic liver injury. Hepatic XBP1s reduces intracellular misfolded protein and regulates lipid and glucose metabolism, and we have demonstrated using murine models that hepatic XBP1s also regulates hepatic bile acid metabolism, although the mechanisms are essentially unknown. The central hypothesis of this grant proposal is that during cholestasis, hepatic XBP1s reduces bile acid synthesis and regulates bile acid metabolism by transcriptionally regulating Cyp7a1, SHP and other bile acid metabolic genes. We present preliminary data demonstrating that acute XBP1s activation transcriptionally regulates hepatic Cyp7a1 to reduce its gene expression, protein expression and function. Therefore, we will determine the mechanisms by which acute liver XBP1s activation transcriptionally regulates Cyp7a1 and the resultant effects on bile acid metabolism (Aim 1). We also provide data demonstrating that XBP1s and FXR both regulate hepatic SHP and will investigate the transcriptional regulation of bile acid metabolism by XBP1s and FXR/SHP and determine the functional consequences (Aim 2). Finally, the bile acid pool and hepatic gene expression differ between mice and humans so we will determine the effects of hepatic XBP1s on human bile acid metabolism using mice with humanized bile acid pools, human hepatocytes and human liver samples (Aim 3). This proposal uses a team of investigators with extensive expertise in transcriptional gene regulation, bile acid metabolism, bioinformatics and medicinal chemistry, combined with state-of-the-art technologies. By determining the hepatic XBP1s regulation of bile acid metabolism, we can identify new molecular targets to develop therapies for cholestatic liver diseases.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY Acute myeloid leukemia (AML) is an aggressive blood cancer that affects roughly three million Americans, and five hundred thousand people die from AML each year. Previous studies indicate that AML is initiated and maintained by a rare population of leukemia-initiating cells (LICs) that are often malignant versions of hematopoietic stem cells (HSCs). Somatic mutations of tumor suppressor gene TP53 in AML are associated with high-risk disease, complex karyotype, resistance to therapies and dismal outcomes. Thus, there is a critical need to identify novel therapeutic targets to improve the treatment outcomes of AML patients with TP53 mutations. We discovered gain-of-function (GOF) mutant p53 proteins frequently found in AML enhance colony formation in vitro and HSC self-renewal in vivo. Utilizing RNA-seq, we observed that p53 mutant HSCs exhibit an enrichment in HSC self-renewal gene signatures. Our ATAC-seq assays showed significant changes in chromatin accessibility in p53 mutant HSCs compared to wild type (WT) HSCs. Our ChIP-seq assays showed that p53 mutant HSCs display high level of repressive histone mark H3K27me3 compared to WT HSCs. Higher-order chromatin structure is emerging as an important regulator of gene expression. Utilizing Hi-C, we discovered that GOF mutant p53 generates specific chromatin loops and alters A/B chromatin compartmentation in leukemia cells, which may contribute to dysregulated gene expression in leukemia cells. Our objective for this research is to determine the role of GOF mutant p53 in the regulation of LIC self-renewal, chromatin accessibility, and 3D chromatin structure in LICs and leukemia cells. The central hypothesis is that specific GOF mutant p53 proteins promote leukemogenesis and mediate therapy-resistance by interacting with transcription regulators that remodel chromatin accessibility and structure in LICs. We will employ multidisciplinary and complementary approaches, including molecular, biochemical, genetic, and pharmacological approaches, as well as unbiased proteomic and genome-wide RNA-seq, ATAC-seq, ChIP- seq, and Hi-C studies to elucidate the mechanisms by which GOF mutant p53 proteins promote leukemogenesis and mediate therapy-resistance. We anticipate that this research will uncover novel mechanisms whereby GOF mutant p53 affects transcription, chromatin accessibility, and 3D chromatin structure in HSCs and AML cells. Therefore, our research will address a significant gap in knowledge regarding the mechanisms by which GOF mutant p53 enhances LIC self-renewal, providing novel therapeutic targets for therapy-resistant TP53-mutant AML.

NIH Research Projects · FY 2026 · 2024-12

Project Summary Primary Ciliary Dyskinesia (PCD) is a rare genetic disorder that affects multiple tissues leading to increased risk for situs inversus, infertility, hydrocephalus and respiratory disease. In the lung, PCD phenotypes are driven by the failure of multi-ciliated cells (MCCs) to generate proper mucociliary flow leading to chronic respiratory infections. To date the combined efforts of many groups have identified over 50 genes causative for PCD. However, the diagnostic tools for identifying PCD patients are time consuming, costly, and phenotypically constrained, likely leading to significant underdiagnosis. Cilia are complex molecular machines consisting of over a thousand proteins that contribute to their motility and function. To function properly, MCCs must: 1.) generate over 100 centrioles that will become the basal bodies of their motile cilia; 2.) dock those centrioles with the apical surface; 3.) nucleate the proper number and length of cilia; 4.) generate proper cilia motility; and 5.) coordinate the timing and orientation of cilia motility. Defects in any of these steps will lead to a decrease in mucociliary flow yet the diagnosis of PCD has been primarily focused on cilia motility. We have developed the ciliated epithelia of the Xenopus embryonic skin as a powerful system to address numerous aspects of ciliary function. Importantly, we have developed quantifiable assays for each step of MCC formation and can identify the underlying cause of a decrease in mucociliary flow. Our combinatorial approach will facilitate a detailed analysis of multiple genes that are either linked to PCD without a clear mechanism (e.g. AK7) or have only been predicted to cause PCD (e.g. Saxo2, Meig1, Rsph14). Additionally, we will perform a tiered CRISPR/Cas9 based genetic screen to identify and functionally characterize novel PCD causative genes. Our goal is to generate an extended list of PCD causing genes that can be included in genetic testing to facilitate the diagnosis of PCD. Additionally, we hope to develop clearly defined subclasses of the disease, marked by loss of motility, loss of cilia number or loss of cilia polarity.

NIH Research Projects · FY 2026 · 2024-12

Project Summary Cellular heterogeneity limits the identification of regulatory networks that control specific cellular functions such as cell secretion. This is particularly challenging for deciphering cellular immunity as immune responses often involve intermediate extracellular signalling acting as autocrine or paracrine modulators. Characterizing such dynamic stimulus-response relationships by analyzing the genes and respective regulatory networks responsible for immune cell secretion at single-cell resolution would be valuable for better understanding and controlling immune responses and would significantly aid drug discovery. However, functional genetic screening required for the identification of genetic regulators of cell secretion dictates sorting large number of cells based on their secretion patterns, which is challenging with existing cell secretion approaches. In addition, performing single- cell analysis of immune cells within a population is critical to accurately decouple autocrine and paracrine secretion cascades, which is not possible via current cell segregation approaches. This proposal will leverage a new microfluidic platform for sorting cells based on their secretion with demonstrated robust performance for the identification of druggable regulators of cell secretion. Our technology – referred to as SECRE (Secretion- Enabled Cell Ranking and Enrichment) – can be used in conjunction with next-generation sequencing to identify the genetic regulators of cell secretion. In this proposal, we will apply the SECRE platform technology for studying multiple types of immune cells and their cell secretion products using a genome-wide CRISPR activation and/or knockout screening. The SECRE assay will be optimized for each cell type and deployed to sort immune cells based on the enhancement of cell secretion. This approach will be used to identify the druggable regulators of IL-2 and CXCL13 secretion by T cells, thus providing an innovative tool for controlling their proliferation and potency. In addition, this approach will be used to explore the druggable regulators of small lytic proteins by NK cells and immunosuppressive cytokine IL-10 by regulatory B cells to enhance their respective potencies, which would provide means to discover new therapeutics for multiple autoimmune diseases. Top screen hits will be selected using well established bioinformatic algorithms and validated through genetic perturbation-based gain-of-function and/or loss-of-function analyses. Using in-vitro functional assays, the cell-type specific secretory networks will be identified and evaluated for potential clinical utility and nominated for further in vivo investigations. The project deliverables will include a first-in-class system for high-performance cell sorting based on their secretory products at single-cell resolution that can be combined with functional genomics to accelerate the development of effective therapeutics for immune disease. This will have immense utility for bioanalysis and/or immunoengineering communities and is expected to enhance drug discovery toolkits. Lastly, this approach will impact the American population by accelerating the development of durable adoptive cell therapies.

NIH Research Projects · FY 2026 · 2024-12

Project Summary The mammalian circadian clock is comprised of an autoregulatory transcription-translation feedback loop expressed in the brain and peripheral tissues that coordinates metabolism with the sleep-wake cycle. Epidemiological and genetic studies have shown that disruption of circadian rhythms leads to accelerated and worsened symptoms of metabolic syndrome. Evidence from skeletal muscle clock mutants in lean mice indicates that loss of clock activators leads to impaired glucose tolerance and reduced insulin-stimulated glucose uptake. Additionally, our lab has shown that genetic abrogation of the skeletal muscle molecular clock in vitro leads to skeletal muscle dysfunction due to reduced mitochondrial oxidative respiration and impaired activation of the hypoxia-inducible factor (HIF) pathway. My preliminary data demonstrate that during the nutrient stress condition of diet-induced obesity (DIO), HIF1α pathway target gene expression is elevated in skeletal muscle and loss of clock activator, BMAL1, leads to reduced HIF1α pathway target gene expression and impaired glucose tolerance in mice. Additionally, the clock-disrupted mice have reduced muscle mass which may be a preliminary sign of sarcopenia. We generated clock-disrupted HIF1α stabilized mice to determine whether this could rescue the muscle phenotypes seen in the clock-disrupted mice. Glucose tolerance and HIF1α target gene expression were rescued in the HIF1α stabilized mice. These data suggest that the skeletal muscle molecular clock regulates glucose metabolism through the HIF pathway, however, the specific mechanisms of this regulatory interaction and the role of HIF1α in maintaining muscle mass remain unknown. The scientific premise of the present proposal is that the skeletal muscle molecular clock controls whole-body glucose metabolism and skeletal muscle metabolism during DIO through regulation of HIF pathway transcription. The studies in this proposal will provide greater insight into clock control of skeletal muscle metabolism during nutrient stress and elucidate the mechanism of interaction between the skeletal muscle molecular clock and HIF pathways.