Northwestern University

NSF Awards · FY 2026 · 2026-06

Generative Artificial Intelligence (AI) tools are increasingly being integrated into creative industries. While generative AI has enormous potential to transform creative work, current tools pose significant risks. For example, use of AI assistants can lead to less innovative ideas, fewer opportunities for skill-development, and reduced feelings of ownership over creative products. To prioritize human creativity and increase human capacity for innovation, this project will design and study a set of generative AI assistants that amplify, rather than replace, human creativity, pushing creative workers to think expansively and develop high quality ideas. This project will achieve this aim by designing tools that employ “generative friction” to foster reflection during the creative process; for example, by critiquing and reframing ideas or generating surprising outcomes. The project will result in a set of novel open-source AI assistants, broadening access to AI that supports creative practice, and novel curricula to build AI literacy among students pursuing creative careers. This work will provide critical insights into how to re-center human creativity as AI is integrated in creative industries. This project advocates for a novel paradigm for designing generative AI assistants for creative practice. The investigators will engage in iterative research-through-design to create a set of four novel AI assistants that leverage generative friction, an oppositional force that emerges from difficulty, surprise, and material or technical constraints that lead a creator to insight, as a mechanism for fostering reflection in creative practice. A within-subjects study will be conducted with 80 digital media artists to investigate how generative friction impacts artists’ creative processes and sense of agency and ownership. In addition, the investigators will conduct a longitudinal diary study with 2 digital media artists to iteratively develop a theoretical model of how generative friction impacts creative choices in long-term naturalistic creative reflective practice with AI. This project will additionally produce curricula to build AI literacy among students in creative careers and a public gallery show of artworks created with the AI tools to expand public discourse surrounding AI and creativity at a critical juncture. 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 · 2026-06

PROJECT SUMMARY/ABSTRACT Psoriasis is a chronic inflammatory skin disease frequently associated with dyslipidemia, characterized by elevated levels of LDL, cholesterol, and triglycerides. While lipid metabolism is implicated in psoriasis pathogenesis, the mechanisms linking dyslipidemia to immune dysregulation remain poorly defined. Recent studies suggests that T cells recognizing self-lipid antigens presented by group 1 CD1 molecules (CD1a, CD1b, CD1c) may act as immunologic sensors of lipid dysregulation, contributing to skin inflammation. However, due to the lack of group 1 CD1 molecules in mice, mechanistic studies of these T cells in vivo have been limited. To address this gap, we developed a double transgenic mouse model expressing human group 1 CD1 molecules (hCD1Tg) and a CD1b-restricted TCR specific for self-phospholipids (HJ1Tg). Crossing these mice onto LDL receptor-deficient (LDLR-/-) background and feeding them a high-fat diet (HFD) induces spontaneous psoriasiform skin inflammation characterized by dermal neutrophil infiltration, keratinocyte hyperplasia, and a Th17-skewed CD1b-autoreactive T cell response. Supporting translational relevance of this model, CD1b expression is elevated in psoriatic skin, and the frequency of circulating CD1b-autoreactive T cells is increased in psoriasis patients. We hypothesize that diet-induced hyperlipidemia chronically activates self-lipid-specific CD1b-restricted T cells and drives IL-17-biased inflammation in the skin. To test this, we propose the following aims: Aim 1: Investigate the mechanisms by which dyslipidemia promotes skin inflammation through CD1b- autoreactive T cells. We will use single-cell RNA sequencing, flow cytometry, and metabolic profiling to define the transcriptional, cytokine, and metabolic programs of CD1b-autoreactive T cells and their interactions with other immune cells under dyslipidemic conditions induced by HFD. Aim 2: Determine the reversibility of dyslipidemia-induced skin inflammation and T cell activation following dietary normalization. Using a diet-switch model, we will examine skin pathology, T cell function, lipid profiles, and gene expression changes in mice transitioned from HFD to normal chow. Aim 3: Evaluate the contribution of gut microbiota to HFD-induced immune dysregulation and skin inflammation. We will utilize antibiotic treatment and metagenomic sequencing to investigate how microbial composition impacts systemic immune activation and CD1b-restricted T cell responses under dyslipidemic conditions. This study aims to define how lipid dysregulation influences autoreactive CD1b-restricted T cell responses, advancing understanding of the immunometabolic mechanisms in psoriasis and related inflammatory diseases. These findings may provide a foundation for targeting lipid-specific T cells as therapeutic interventions for dyslipidemia-associated skin inflammation.

NIH Research Projects · FY 2026 · 2026-06

Project Summary: Genetic heart disease is associated with heart failure and arrhythmias, which can lead to significant morbidity and mortality. Small animal models of genetic heart disease often fail to capture the clinically relevant features of the disease. Patient- and gene-specific therapies, such gene therapy, gene editing, and exon skipping are making their way into the clinic and are in development. In many cases, genetics has provided a clear understanding of underlying patient-specific disease mechanisms by linking a patient’s disease to a specific gene variant. However, individualized therapeutic strategies lag behind this understanding. Recognizing the limitations of animal models, the FDA Modernization Act 2.0 (2022) and subsequent FDA regulatory guidance (2024) now allow for the use of non-animal models including cell- and organoid-based models to assess therapeutic and efficacy. Patient-specific human induced pluripotent stem cells (hiPSCs) can now be readily created from patient cells such as those obtained via a standard blood draw. These cells can then be differentiated in heart-like cells to created hiPSC-derived cardiomyocytes (hiPSC-CMs), offering an platform to test personalized, genotype-specific therapies for heart disease in vitro. HiPSC-CM models can be further improved by using them to create heart-like tissues in the dish, known as engineered heart tissues (EHTs). Commercial platforms now support the creation of these tissues, and contractility measurements can be obtained non-invasively, enabling therapeutic assessment. However, arrhythmia assessments in EHTs remain limited due to the need for specialized optical equipment, the use of toxic contraction inhibitors such as blebbistatin, the need for voltage-sensitive dyes, and the terminal nature of current experimental protocols, which restrict the ability to track therapeutic effects over time. We have recently developed an electromechanically monitored EHT (emEHT) platform that enables simultaneous measurement of contractility and field potentials. This platform leverages flexible electronics technology to noninvasively and concurrently detect electrical signals and assess tissue forces. We have previously demonstrated the ability to simulate arrhythmias using electrophysiology protocols adapted from the clinic with this system. This proposal aims to develop a next-generation emEHT platform capable of simultaneously assessing action potentials and calcium transients in a non-invasive, non-terminal format through the integration of flexible electronics embedded with optical microsensors. Additionally, we aim to develop the molecular tools necessary to leverage this emEHT platform, including the stable expression of genetically encoded voltage and calcium sensors. The platform will be validated against traditional optical mapping techniques and tested using hiPSC-CMs derived from an arrhythmic form of cardiomyopathy. The development of this platform holds transformative potential for assessing arrhythmia propensity in EHTs, ultimately enabling the evaluation of the functional consequences of genotype-specific cardiovascular therapeutics in the dish.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT Alternative splicing vastly expands the functional repertoire of genes, and is especially important during human synapse development. Human synapses have unique structures and compositions, but the mechanisms involved in their development remains elusive. Investigations into human-specific aspects of synaptic machinery have revealed a critical role for Rho guanine nucleotide exchange factor (GEF) signaling. TRIO is a large multi-domain RhoGEF with essential roles in neuronal and synaptic development. TRIO is also a genome-wide significant risk factor for neurodevelopmental disorders and glutamatergic synapses have been implicated by genomic, neuropathological, and functional studies as key sites of pathogenesis. Individuals with pathogenic TRIO variants are characterized by intellectual disability, developmental delay, and seizures. The TRIO gene produces multiple distinct isoforms that differentially incorporate TRIO’s functional domains including two GEF domains that having opposing functions on the actin cytoskeleton. Thus, precise control of TRIO’s GEF domains through expression and localization of isoforms is paramount for its function. However, the diversity and expression patterns of TRIO isoforms are poorly understood, and their impact on human synapse development is unknown. A complete catalogue of isoform diversity is necessary for a holistic view of gene function, and is critical for gene therapy designs, interpreting clinical variants, and revealing novel biological mechanisms. Here, we will leverage state- of-the-art long-read sequencing, iPSC-models and fluorescence imaging to characterize human TRIO isoforms. Our central hypothesis is that TRIO produces an array of isoforms with distinct expression patterns and domain architectures that assist with the precise timing and spatial control of synaptic development. In Aim 1, we will combine exon capture and long-read sequencing technologies to systematically profile full-length TRIO transcripts in the human brain. We will create comprehensive isoform maps at three postnatal time points, and analyze novel isoforms for differential expression, presence of clinical variants, and protein motifs. In Aim 2, human iPSC-derived neuron models will be utilized to assess the localization and functional roles of individual isoforms in synaptic compartments. Together, this work will reveal a high-resolution map of the structure, expression and function of TRIO isoforms in the human brain, providing mechanistic insight into human synapse development and TRIO-related neurodevelopmental disorders.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Animals depend critically on their capacity to choose the appropriate time to act. In the absence of explicit cues, these decisions are thought to depend on internal deliberation that dictates action timing to maximize outcomes. A broad range of previous observations suggest that internally driven timing decisions involve a distributed network of brain regions. However, it remains unclear how these regions interact during decision-making; mechanistic understanding is very limited and hypothesized interactions between brain regions remain untested. This critical knowledge gap stems from two main factors. First, the absence of explicit cues makes decision timing unpredictable, which historically has presented problems for experimental design. Second, previous methods for measuring and perturbing neural activity, and for quantifying the dependence relations between neural activity patterns, have been ill-suited for probing interactions between brain areas on the relevant timescales. To overcome these barriers, our collaboration combines a novel behavioral paradigm, multi-region spike- resolution neural recording, rapid neural activity perturbation, and a range of model-based computational approaches. In recent work, our analysis of activity on individual decision trials has demonstrated the prominent involvement of a deterministic process, in contrast to recent models that emphasize stochasticity. We have also developed an approach for analyzing multi-region recordings that has revealed a modular structure in the influence of several frontal cortical regions on the striatum, an influence thought to be central to timing decisions. Our preliminary results for this proposal point to a revised decision model that involves an urgency-like signal and a source of unpredictability distinct from that of prevailing models. We have also begun to examine interactions between prefrontal and somatomotor circuits that are also thought to be central to timing decisions. Here we have found evidence of an interaction mediated primarily by corticocortical connections, and one that has a modular structure. Our proposed work would build on these results to test our new model and identify its neural substrates (Aim 1), and test existing ideas about prefrontal-somatomotor interactions (Aim 2). Here we will use multi-region Neuropixels recordings, and an approach we have recently demonstrated for fast optogenetic silencing during internally driven timing decisions. We will compare our new model to others using rigorous statistical methods for model selection. We will identify neural substrates by analyzing best-fit models and perturbation results. We will also continue to develop our new methods for quantifying interregional interactions from activity recordings. Collectively, our work will quantify relevant interactions between brain regions that could not be resolved with previous approaches, leading to improved models. This will provide a new foundation for understanding the neural mechanisms of a basic aspect of natural behavior with relevance to cognition.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Viral pneumonia represents the leading infectious cause of death in the United States. In almost all patients, severe pneumonia presents as the acute respiratory distress syndrome (ARDS), characterized by an exuberant immune cell-mediated response that injures the alveolar epithelial-capillary barrier resulting in impaired gas exchange and respiratory failure. In patients with viral pneumonia, we reported that the persistence of inflammatory signaling circuits between hyperactivated T cells and macrophages in the alveolar space is associated with immunopathology, dysregulated lung repair, and poor clinical outcomes. FOXP3+ regulatory T (Treg) cells are known to infiltrate both human and murine lungs to orchestrate lung repair after viral pneumonia through expression of suppressive mediators. Key among these suppressive factors is the anti-inflammatory cytokine Interleukin (IL)-10. My published work supports a key role for pro-resolving Treg cell transcriptional program enriched for IL-10 expression during successful recovery from viral pneumonia. During recovery from viral pneumonia, chronic inflammatory infiltrates are spatially organized into structured lymphoid aggregates known as inducible bronchus-associated lymphoid tissue (iBALT). While this form of ectopic lymphoid tissue has been causally linked to outcomes in murine models of COPD and lung transplant rejection, their role as a marker of, or cause of, failed lung repair during viral pneumonia is not known. My preliminary data using spatial transcriptomics demonstrates that iBALT is disproportionately expanded in old compared with young mice after viral pneumonia. Intercellular IL-10 signaling analysis revealed old mice exhibited dampened inter-niche connectivity of the IL-10 gene program. Transcriptional profiling of immune cell subsets from over 70 bronchoalveolar lavage (BAL) human samples revealed that Treg cells are the dominant source of IL-10 in the alveolar space and their enrichment associates with outcomes. We hypothesize that Treg cell generated IL-10 signals to monocyte-derived alveolar macrophages (MoAM) and interstitial macrophages (IM) to interrupt inflammatory circuits between MoAM/IM and activated effector T cells, limiting iBALT formation and promoting recovery from viral pneumonia. We propose three Specific Aims, which use innovative murine systems, a translational human case-control study, and cutting-edge computational procedures to test our hypothesis. Aim 1 will determine whether IL-10 production by Treg cells is necessary and sufficient to mitigate iBALT formation and accelerate recovery from viral pneumonia. In Aim 2 we will determine whether signaling through the IL-10Ra in MoAM and IM is necessary for Treg cell-dependent mitigation of iBALT formation during recovery from viral pneumonia. Aim 3 will ascertain whether IL-10+ alveolar Treg cell-specific transcriptional programs and Treg cell receptor (TCR) profiles associate with outcomes in patients with severe viral pneumonia.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Magnesium (Mg2+) is essential for brain and nervous system health, playing critical roles in neuronal excitability, synaptic transmission, and protection against excitotoxicity. Disruptions in Mg2+ homeostasis— particularly chronic hypomagnesemia—are associated neurological disorders such as epilepsy, developmental delay, and neurodegeneration. The CNNM (Cyclin and CBS Domain Magnesium Transport Mediator) family of membrane proteins plays a central role in maintaining cellular and systemic Mg2+ balance. Mutations or dysfunctions in CNNMs are linked to severe neurological phenotypes, underscoring their importance in nervous system function. The CNNM family includes four members (CNNM1–4), each with distinct tissue distribution and physiological roles. These evolutionarily conserved proteins mediate Mg2+ efflux from cells, thereby regulating Mg2+ homeostasis critical for neuronal signaling and synaptic function. Despite their neurological relevance, the molecular architecture and transport mechanism of full-length CNNMs remain entirely unknown. Fundamental questions persist regarding how CNNMs recognize MgATP, undergo conformational changes to mediate Mg2+ efflux, and couple these processes to dynamic intracellular domains. Additionally, CNNM activity is modulated by small-molecule compounds and regulatory proteins such as PRLs (Phosphatases of Regenerating Liver) and ARL15 (ADP-Ribosylation Factor-Like GTPase 15), but how these factors alter transporter structure and function at the molecular level is poorly understood. We propose to define the structure and transport mechanism of CNNMs by integrating high-resolution cryo- electron microscopy with biochemical and functional assays. These studies will reveal how CNNMs transition through distinct functional states, how they sense intracellular Mg2+ levels, and how allosteric regulators— including small-molecule compounds and regulatory proteins—modulate their activity. The results will establish a comprehensive molecular framework for CNNM-mediated Mg2+ homeostasis and provide the foundation for future therapeutic strategies targeting magnesium-related neurological disorders.

NIH Research Projects · FY 2026 · 2026-06

Dementia with Lewy bodies (DLB) and Parkinson’s disease (PD) are characterized by the accumulation of proteinaceous inclusions within the nervous system. Genetics has indicated that protein homeostasis, mitochondria, and protein trafficking are major pathways that contribute to disease. We recently discovered that proteostasis failure in DLB and PD results in the accumulation of nuclear inclusions comprised of RNA binding proteins (RBPs) NONO and SFPQ. Pathologically, these inclusions increase adenosine-to-inosine (A-to-I) RNA editing by reducing the transcription of ADAR3, the inhibitor of A-to-I RNA editing. NONO and SFPQ also preferentially sequester A-to-I edited mRNAs. Our previous work showed that A-to-I editing can increase the binding to NONO/SFPQ inclusions, and further template their aggregation. Bulk RNA sequencing showed that increased editing occurs in over 3000 sites in 1600 transcripts in synucleinopathy patient iPSC-derived neurons. Among the top networks were transcripts encoding mitochondrial, ER-Golgi trafficking, or axon/synaptic maintenance proteins, which overlaps with PD/DLB genetic pathways and pathophysiology. However, the composition of mRNAs sequestered by NONO/SFPQ inclusions and the initial events that trigger inclusion formation is unknown. Furthermore, the downstream effect that A-to-I editing has on protein expression and function is unknown. Here we will address these critical questions in 3 aims using both iPSC-derived cortical neurons to model dementia-related phenotypes, and midbrain neurons to model both early and late stages of DLB, PD, and PD-Dementia. Studies will be validated in post-mortem Dementia with Lewy body patient brain. Aim 1 will determine how A-to-I editing influences subcellular location of RNA and define the specific mRNAs sequestered within NONO/SFPQ inclusions. Aim 2 will assess the connection between mitochondrial and ER- Golgi stress and NONO/SFPQ/RNA inclusion formation. Aim 3 will delineate the downstream effects that A-to-I editing has on protein expression and function by Ribosome foot-printing and prioritizing select model transcripts involved in mitochondrial, ER-Golgi trafficking, or axon/synaptic maintenance. Our studies will contribute to delineating the mechanism of A-to-I editing in synucleinopathies, and link the editing changes we previously observed to established genetic pathways associated with DLB and PD. Our mechanistic studies may uncover new methods to clear NONO/SFPQ aggregates and restore neuronal health in dementia patients. Since A-to-I editing and paraspeckles are both involved in controlling the expression of synapse/axon maintenance proteins, are studies may apply to many types of neurological disorders characterized by early synapse loss.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY This proposal seeks to acquire a RH96 Octet – a next-generation, high-throughput bio-layer interferometer (BLI HTS) for biomolecular interaction studies. BLI HTS is a “96-channel” instrument that allows quantitative measurements of protein-protein and protein-small molecule interactions of up to 96 samples simultaneously. It uses small sample volumes, and the ease of use allows researchers with broad scientific expertise to use the instrument for high-throughput screening of interactors such as proteins, antibodies, and small molecules. There is an urgent unmet need for this instrument, as there is no high-throughput BLI instrument in the entire Midwest region. We have identified 14 NIH-funded researchers whose work and progress is severely limited due to the absence of this instrument. They often travel to out-of-state institutions or use the 2-channel BLI instrument currently available at Northwestern; clearly, neither option is a long term solution. The 2-channel instrument is not amenable to high-throughput data collection and requires large sample volumes, impeding drug discovery and screening efforts. Moreover, the data collection on the 2-channel is tedious even for single protein-protein interaction studies when testing multiple concentrations. The proposed instrument will address a significant gap in the instrumentation available at Northwestern University and will be beneficial not just to the Northwestern community but the entire Midwest area. We propose to add the BLI HTS to the Northwestern High-throughput Analysis Laboratory, which houses several instruments for high-throughput screening and has full-time staff with the technical expertise to manage and operate the instrument. We anticipate that at least 14 research groups from 6 departments across the College of Arts and Sciences, the Feinberg School of Medicine and the McCormik School of Engineering will utilize this equipment. The availability of this state-of-the-art instrument at Northwestern will be particularly important for drug development efforts and will advance a range of studies aimed at creating therapeutic and diagnostic tools for various human diseases, including cancer, metabolic disorders and neurodegeneration.

NIH Research Projects · FY 2026 · 2026-06

Project Summary Among the 18.1 million U.S. cancer survivors, approximately 40% experience physical and functional impairments (PFIs) due to cancer and its treatment. PFIs are frequently undertreated, contributing to functional disability in 10–25% of survivors. They negatively affect health-related quality of life (HRQoL), interfere with treatment completion, drive costly acute care utilization, and may impact survival. Despite the proven benefits of physical, occupational, and speech therapy, cancer rehabilitation services remain underutilized (e.g., often initiated only after impairments become chronic). Cancer survivors are ill-equipped to self-manage PFIs or navigate the healthcare system to access appropriate services, and oncology teams lack tools for timely detection and referral. Health systems also lack decision support tools to match patients to the appropriate rehabilitation specialty and nearby clinic locations. Timely identification and treatment of PFIs is essential to improving outcomes and reducing avoidable disability, and both symptom and financial burden. To address these gaps, we developed the 4Rs Cancer Rehabilitation Triaging System: a novel, EHR-integrated platform that identifies the Right patient at the Right time and matches them to the Right rehabilitation specialty in the Right location. By embedding systematic PFI monitoring and triaging into routine oncology care, 4Rs reduces clinician burden while supporting early intervention and patient self-management. This R01 will conduct a two-arm randomized controlled trial to evaluate the clinical and implementation outcomes of 4Rs across oncology clinics in a large healthcare system (Northwestern Medicine). Aim 1 will assess the efficacy of 4Rs among 1,000 cancer survivors with PFIs identified within three months of diagnosis, compared to usual care. Participants will be followed for two years. The primary outcome is PFI measured by PROMIS-PF. Secondary outcomes include urgent/emergency care utilization, HRQoL, and survival. Aim 2 will evaluate (a) the pre-implementation process and cost effectiveness of 4Rs using the PRISM/RE-AIM framework to assess reach, adoption, implementation, and maintenance via surveys and focus groups with patients, clinicians, and administrators (n=60); (b) cost effectiveness of 4Rs that includes potential implementation and maintenance costs as well as cost savings. Aim 3 will evaluate mechanisms of action, examining intervention targets, mediators (e.g., patient self-efficacy), and moderators (e.g., cancer type, treatments received) that influence 4Rs outcomes. By improving access to timely, tailored cancer rehabilitation, this project has the potential to transform survivorship care, reduce disparities in rehabilitation access, and inform scalable models for value-based cancer care delivery.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT Recent advances have uncovered important mechanistic insights regarding how LKB1/STK11 functions as a tumor suppressor in the lung, yet understanding of LKB1 mutant lung tumor biology, and how to successfully treat this devastating disease, remains incomplete. Our cancer biology studies have revealed that HDAC3 is essential for LKB1 mutant lung tumor growth, and that HDAC3 is a druggable vulnerability in therapeutic resistance to KRAS pathway inhibitors. However, a knowledge gap remains in our mechanistic understanding of how LKB1 loss regulates HDAC3/NKX2-1 complex function in lung tumors, and how KRAS pathway inhibitors impinge on this molecular node. Our proposal aims to bridge this knowledge gap by (1) determining how LKB1 specifies the HDAC3/NKX2-1 program in lung cancer cells, (2) testing if LKB1 status dictates whether KRAS pathway inhibition and entinostat combination treatment controls lung tumor growth, and (3) defining how KRAS pathway inhibitors impact HDAC3/NKX2-1 to regulate lung tumor growth. We will use genomics and mass spectrometry approaches coupled with lung cancer cell lines and mouse models of lung cancer to achieve these goals. The use of mouse models for a limited set of experiments is necessary in order to accurately understand the LKB1 pathway, as this pathway is established to function differently in vitro compared to in vivo. Moreover, mice are essential for understanding the impact on tumor growth control in the context of an intact immune system. Overall, these studies will advance our understanding of the genetic and therapeutic mechanisms modulating HDAC3 complex function in lung tumor biology, and pave the way toward improved therapeutic options for patients with LKB1 mutant lung cancer.

NIH Research Projects · FY 2026 · 2026-06

Project Description When parents engage in their adolescents' substance use (SU) treatment, the adolescents experience better outcomes and lower risk of relapse. Yet, parent engagement remains low across the SU treatment continuum, particularly in service lines designed for adolescents with severe SU. To address this gap, we created Parent SMART – a scalable technology-assisted intervention (parent networking forum + online program + telehealth coaching sessions) – to engage parents in adolescent residential SU treatment. Through a successful NIDA- funded R34 and the first segment of this R37, we established Parent SMART's feasibility, acceptability, and preliminary effectiveness on parenting processes, adolescent SU, and adolescent school-related problems. We also showed our ability to conduct a pragmatic effectiveness trial and efficiently integrate Parent SMART into two of the largest adolescent residential facilities in the country: Rosecrance Griffin Williamson and Hazelden Betty Ford. With the enthusiastic support of our partners, we now propose to adapt Parent SMART to reflect a changing national treatment landscape. Since 2010, the numbers of residential programs for youth with SU and other behavioral health problems, youth served, and available beds have declined by 61%, 78%, and 66%, respectively, while the use of crisis hotlines, outpatient wraparound services, and intensive outpatient services has surged. This shifting service mix highlights the need to proactively enhance parent engagement across the SU continuum of care. In this next R37 segment, we will adapt and implement Parent SMART to ensure that it fits the needs of parents, adolescents, and staff in those service lines experiencing a surge in utilization. Parent SMART adaptation will be guided by ADAPT-ITT, an 8-step, systematic process to adapt evidence-based interventions. In the first six steps of ADAPT-ITT, we will identify core intervention elements that transcend settings and solicit constituent feedback to adapt modifiable, peripheral elements (Aim 1). In the final two steps of ADAPT-ITT, we will employ a hybrid type 2 effectiveness-implementation trial to simultaneously evaluate the adapted Parent SMART intervention and its implementation (Aim 2). Parent SMART implementation will be guided by the Science-to-Service Laboratory, a three-tiered (didactic workshop + performance feedback + coaching) strategy that has demonstrated effectiveness across several of PI Becker's clinical trials. To harmonize data collection across R37 segments, we will use the same measures of patient effectiveness (parental monitoring, parent-adolescent communication, adolescent SU) and assessment calendar (baseline, 12-, and 24-weeks). Implementation outcomes will include reach, adoption, and fidelity, following guidelines of the NIDA-funded Research Adoption Support Center, to align with other research networks. This work will advance the field by: targeting a high-need population wherever they access services; removing barriers to parent engagement; testing the adapted intervention and its implementation simultaneously to accelerate impact; and informing development of a Parent SMART commercialization and sustainment plan. D UEI: KG76WYENL5K1 Enter name of Organization: D RESEARCH & RELATED BUDGET - Budget Period 1 OMB Number: 4040-0001 Expiration Date: 11/30/2025 Northwestern University Budget Type: Project ? Subaward/Consortium Budget Period: 1 Start Date: 06/01/2026 End Date: 05/31/2027 A. Senior/Key Person Months Requested Fringe Funds Prefix First Middle Last Suffix Base Salary ($) Cal. Acad. Sum. Salary ($) Benefits ($) Requested ($) Sara Becker 221,900.00 2.40 44,380.00 12,471.00 56,851.00 Project Role: PD/PI D D Sarah Helseth 148,447.00 3.00 37,112.00 10,428.00 47,540.00 Project Role: Co-Investigator D Zabin Patel 109,273.00 1.20 10,927.00 3,071.00 13,998.00 Project Role: Co-Investigator Additional Senior Key Persons:Add Attachment Delete Attachment View AttachmentTotal Funds requested for all Senior Key Persons in the attached file Total Senior/Key Person 118,389.00 B. Other Personnel Number of Personnel Project Role Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical n n n n 2 1 n Research Assistants Project Manager 3 Total Number Other Personnel Months Requested Fringe Funds Cal. Acad. Sum. Salary ($) Benefits ($) Requested ($) D D D D 24.00 D 6.00 D D D D D D D D D D 99,000.00 39,784.00 27,820.00 11,179.00 126,820.00 50,963.00 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) n 177,783.00 296,172.00 n

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY In genetic medicine, variants of uncertain significance (VUS) have posed a significant challenge. These variants, identified through genetic testing, are alterations in one's DNA sequence that cannot be definitively classified as benign or pathogenic. Approximately 80% of missense variants in the ClinVar database are classified as VUS. In clinical care, a VUS result frequently prevents clinicians from providing definitive diagnoses, limits treatment options, and causes uncertainty for patients and their families. Resolving VUS is thus essential to improve patient care in genetic medicine. Multiplexed Assays of Variant Effect (MAVEs) offer a high-throughput solution to VUS resolution by assessing variant effect for thousands of variants in vitro simultaneously. However, there are two limitations to realizing the potential of MAVEs to resolve VUS at scale. First, it is challenging to introduce a comprehensive set of variants into cells at scale, and second, MAVEs have historically relied on a small set of function assays, such as proliferation or survival-based assays, limiting the number of genes that can be investigated. This proposal aims to address both of these limitations, using CHD2 as a model gene, but developing techniques that could be applied to hundreds of clinically relevant genes. We will first advance high-throughput endogenous genome editing through the development of a suite of computational and experimental tools for prime editing. While prime editing offers a precise and versatile approach, it is plagued by low editing efficiencies. To address these shortcomings, we will develop a computational pipeline to design prime editing guide RNA (pegRNA) libraries with high predicted editing efficiency, an optimized cell line for editing, and a FACS-based approach to enrich for edited cells. We expect that such tools will boost prime editing efficiency for multiplexed cell libraries and thereby increase the efficiency of MAVEs. We will also address the limited set of functional assays for MAVEs. Here we propose scAVER-seq: a widely applicable and scalable single-cell Assay of Variant Effect using RNA-sequencing. This technique will combine transcript capture and long-read technology in single cells to identify each cell’s introduced variant and link it to its corresponding transcriptome in a single-tube workflow. By using the transcriptome as a functional readout, we will revolutionize variant classification in disorders associated with robust transcriptional effects, such as those caused by pathogenic variants in regulators of gene expression. This approach could help resolve the over 500,000 VUS that exist in gene expression regulators, which comprise 31% of all VUS in ClinVar. By developing generalizable methods to characterize VUS, we aim to contribute to the NHGRI's mission of understanding the impact of genomic variation on human health and simultaneously help deliver answers to families facing genetic testing results reporting VUS.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract This application is to purchase an AxL Cleared Tissue LightSheet (AxL CTLS) from 3i for imaging of optically cleared samples. The Center for Advanced Microscopy is the only microscopy based core facility on the Chicago Campus and supports approximately 750 users from 300 laboratories annually. The Center for Advanced Microscopy has been supporting light sheet microscopy for 8 years with a Miltenyi UltraMicroscope II system. Light sheet microscopy has been very successfully incorporated into the workflows of many researchers at Northwestern University. Our UltraMicroscope II has been heavily used, but it has reached end-of-life and will no longer be supported by the manufacturer. The instrument is beginning to fail and in the past year we had 4 months of downtime for repairs. We seek to replace this instrument with a new light sheet that has higher resolution, faster speeds and can accommodate a wider range of samples. After testing several systems, we chose the AxL CTLS. Fifteen NIH-funded investigators are Major Users of this instrument and the success of their 26 NIH funded research is dependent on light sheet technology being available at CAM. Their research has a broad range of impacts across basic and applied science fields from fields including surgery, infectious diseases and microbiology, immunology, cardiovascular biology, developmental biology, neurosciences, and nephrology. Acquisition of the AxL CTLS would enhance the research of these investigators and offer improved image capabilities than the current instrument. The AxL CTLS would be a valuable acquisition not only for research carried out at Northwestern University and the Chicago scientific community.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Pantothenic acid, also known as Vitamin B5 (VitB5), is a member of the vitamin B family found in diets and gut microbiota and is primarily absorbed by the intestines. VitB5 is a precursor for the biosynthesis of coenzyme A (CoA). As the major carrier of activated acyl groups within cells, CoA is critical in various core metabolic processes, including nutrient catabolism and lipid synthesis. VitB5 has been shown to regulate the functions of different cells and various diseases. Isolated VitB5 deficiency results in metabolic imbalance and gastrointestinal symptoms. A deficiency in VitB5 was recently reported in the blood and feces of IBD patients; accordingly, VitB5 administration enhanced intestinal barrier repair, suggesting a potential correlation between VitB5 reduction and IBD development. However, how VitB5 regulates IBD is still largely unknown. Our preliminary data showed a lower level of VitB5 in the serum of IBD patients and lower expression of pantothenate kinase (PANK), one of the enzymes that catabolize VB5 to CoA, in Foxp3+ Tregs in IBD patients, compared with healthy controls. This suggests a potential role of VitB5 in the regulation of IBD development through Tregs. We further showed that supplementation of VitB5 protects intestinal inflammation with higher levels of Foxp3+ Tregs in the intestines. This indicates a crucial role for VitB5 in regulating Treg and ongoing intestinal inflammation by promoting Foxp3+ Tregs. Thus, the central hypothesis of this project is that VitB5 protects the intestines from inflammation by promoting Treg suppressive function in the intestines. VitB5-SLC5A6 axis induces Foxp3+ Tregs through increased mitochondrial oxidation and epigenetic regulation, limiting intestinal inflammation progression in IBD. We will test our hypothesis in this application to determine (1) the impact of the VitB5-Slc5a6 axis in Tregs in the regulation of colitis.; (2) whether VitB5 promotes Treg function by enhancing Treg stemness; and (3) the mechanisms by which VitB5 promotes Treg stemness and the protection of intestinal inflammation.

NIH Research Projects · FY 2026 · 2026-05

Various neurodegenerative disorders including frontotemporal dementia (FTD), Alzheimer’s Disease (AD), progressive supranuclear palsy (PSP), and others share a common pathologic feature involving the deposition of abnormal tau protein in the brain (tauopathies). This implies that there may be some shared pathophysiologic mechanism(s). The primary risk factor for nearly all of these conditions is aging, implying that the aging process plays a role in their common pathophysiology. Numerous changes in the aging brain such as compromised autophagy, loss of proteostasis and mitochondrial dysfunction have been implicated in the pathophysiology of these diseases, but no specific molecular change has been identified that leads to disease onset and progression. There is an age-related increase in bone morphogenetic protein (BMP) signaling in both mouse and human brain, and reducing BMP signaling in aging mice reduces aging-related changes in both neurogenesis and cognition. BMP signaling increases levels of phosphotau (p- Tau) in vivo in mouse brain and in vitro in human excitatory neurons derived both from FTD (MAPT mutation) iPSC and from AD iPSC. Conversely, treatment with the BMP inhibitor, noggin, reduces tau phosphorylation and release of p-tau. These studies in vitro provided a strong rationale for examining effects of inhibiting BMP signaling in vivo. Remarkably, noggin overexpression prevented tau hyperphosphorylation, neuropathological changes, and behavioral abnormalities in the P301S tauopathy mouse model and similarly inhibited tau phosphorylation, pathological changes and behavioral abnormalities in mice with knockin of human APOE4. Thus, evidence from two mouse models, iPSC-derived human neurons, and postmortem analysis of human brains strongly supports the hypothesis that interventions that reduce BMP signaling in aging brain could potentially slow or prevent development of disease. The goal of the proposed preclinical studies is to develop a preclinical approach for overexpression of noggin that could be practically translated clinically.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract This application is to purchase a Transmission Electron Microscope (TEM) for the Center for Advanced Microscopy (CAM) at Northwestern University’s Feinberg School of Medicine, which has been supporting TEM with a FEI TECNAI Spirit G2 since 2006. TEM has been essential for the workflows of many researchers at, from fields including surgery, infectious diseases and microbiology, immunology, cardiovascular biology, developmental biology, neurosciences, nephrology, and ophthalmology. CAM’s TEM is heavily used, but is now reaching end-of-life. In the past year alone, it has faced major disruptions due to camera failures, and as a result the image quality that researchers are able to acquire has greatly deteriorated. Sixteen NIH-funded investigators need this instrument at CAM to support their 33 NIH funded projects. The current instrument has been essential for their research as demonstrated by their multiple publications in both basic and applied science fields. The acquisition of a new TEM would enhance the research of these investigators and others, ensuring that they can continue to do their work with the highest quality instrumentation.

NIH Research Projects · FY 2026 · 2026-05

Summary The immune co-receptor protein family plays an important role in fine-tuning immune responses. Previous studies have primarily focused on the stimulatory and inhibitory co-receptors expressed by different T cell subsets and their modulatory effects on T cell activation. Inhibitory co-receptors have recently gained significant attention due to their potent ability to reinvigorate T cell responses in the context of chronic infection and cancer. Besides T cells, myeloid lineage cells can express inhibitory co-receptors, and it has been suggested that these receptors may control their activation, potentially interfering with anti-tumor immunity or contributing to neurological disorders. Lymphocyte activation gene 3 (Lag3) is a CD4-like inhibitory co-receptor expressed on various types of T cells, including activated, exhausted, and regulatory T cells. Lag3 is considered a therapeutic target for immune checkpoint blockade to bolster T cell effector functions. We recently reported that Lag3 plays a crucial role in Treg cell functions. Utilizing newly developed Treg cell-specific Lag3-mutant mouse models, we demonstrated that Lag3 expression is essential for adequate Treg cell function to control experimental autoimmune encephalomyelitis (EAE), an autoimmune inflammation in the central nervous system. From analyzing a published single cell RNA sequencing dataset that compare CNS cells between healthy mice and mice with EAE, we unexpectedly found that microglia express high levels of Lag3 even at steady state condition and that the expression level further increases in EAE. Microglial Lag3 expression has previously been proposed to regulate microglia activation in neurological disorders. However, its role in autoimmune neuroinflammation remains explored. In this application, we will use microglia-specific Lag3-mutant mouse models to test the hypothesis that Lag3 regulates microglial functions by promoting their differentiation into anti-inflammatory subsets, thereby modulating autoimmune inflammation. The hypothesis will be tested by the following two specific aims. Aim 1 will test whether microglia Lag3 deficiency influences microglia maintenance at steady state condition and autoimmune disease development. Aim 2 will investigate the mechanism by which Lag3 controls microglial differentiation and functions. The results from this proposal will open new opportunities to investigate the cellular mechanisms through which microglia activation can be modulated by Lag3. Successful completion of this study will identify novel therapeutic opportunities to regulate microglia functions by targeting Lag3.

NIH Research Projects · FY 2026 · 2026-05

Project Summary Primary mitochondrial diseases (PMDs) are the most common inborn errors of metabolism, affecting ~1 in 5,000 births with devastating neurologic manifestations. A hallmark clinical feature of PMD is infection-triggered permanent neurologic deterioration, where intercurrent infections precipitate acute worsening of neurologic function without return to baseline after infection resolution. This distinctive pattern provides a compelling model to investigate fundamental mechanisms underlying post-infectious neurologic sequelae applicable to other infection-associated chronic neurologic illnesses and post-viral fatigue syndromes. Emerging preclinical and patient data support a central role for the immune system in the progression of PMD. Based on these emerging data, we propose that infections causally accelerate neurodegeneration in PMD through immune cell intrinsic mitochondrial dysfunction that results in dysfunctional immunity, and that different genetic causes of PMD lead to distinct patterns of immune alterations that actively contribute to the severity and phenotype of post-infectious neurologic deterioration. We aim to define the spectrum of immune dysfunction across diverse genetic forms of human PMD through comprehensive immunophenotyping and functional assessment using single-cell RNA sequencing and cytokine profiling of patient peripheral blood mononuclear cells at baseline and post-stimulation. Further, moving beyond correlation, we will determine if mitochondrial dysfunction in immune cells causally accelerates neurodegeneration using the NDUFS4 knockout mouse model of PMD. Specifically by investigating whether influenza infection drives neurologic decline and leveraging genetic restoration of mitochondrial function specifically in immune cells to establish causality. This work will integrate translational pipelines including single- cell sequencing with novel animal models to causally define the mechanisms linking mitochondrial dysfunction, immune dysregulation, and post-infectious neurologic deterioration. Importantly, This research directly addresses NINDS priorities by investigating mechanisms of mitochondrial dysfunction contributing to nervous system dysfunction in infection-associated chronic illnesses, providing insights into shared pathways across multiple post-infectious neurologic conditions and identifying potential therapeutic targets for these disorders.

NIH Research Projects · FY 2025 · 2026-05

PROJECT SUMMARY Light is a ubiquitous feature of the natural environment and plays a central role in modulating an organism’s internal state. This flexible adaptation of neural circuits is essential for an organism’s survival. It endows them with not only the ability to preemptively adapt neural circuits to rhythmic, predictable changes in light levels, but also to reactively adapt circuits to acute, unpredictable changes in light levels. Decades of research have established a neural circuit through which light flexibly modulates internal state based on daily rhythms in light. However, the mechanism through which light acutely tunes internal state is currently unknown. This proposal aims to identify a mechanism through which light can acutely influence a wide range of neural circuits. Retinal ganglion cells are the output cells of the retina and send information to over 50 areas in the brain. Among these regions which receive direct retinal input is the basal forebrain. The basal forebrain houses the acetylcholine neuromodulatory system, which sends information throughout the brain to acutely influence a wide range of behaviors. The basal forebrain is known to modulate numerous behaviors including attention, arousal, sleep/wake, and learning and memory. Notably, all these behaviors are also modulated by light. Thus, the retinal input to the basal forebrain is a strong candidate for the acute modulation of behavior based on environmental light. This proposal will identify the types of visual information which are relayed to the basal forebrain, the basal forebrain circuits and cell types this information is integrated into, and how this contributes to light modulation of behavior. In Aim 1, I will identify the retinal inputs to the basal forebrain and define the neural circuits this visual information is integrated into using virus-mediated circuit tracing and electrophysiology. In Aim 2, I will determine how retinal input to the basal forebrain influences a basal forebrain dependent behavior. Collectively, the proposed experiments will identify retinal inputs to one of the key neuromodulatory centers in the brain, providing insight into the mechanism through which light can acutely modulate a wide range of behaviors.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Epilepsy is a major source of death and disability worldwide. Specialists diagnose epilepsy by integrating the clinical history with specialized tests, particularly the electroencephalogram (EEG). Accurate diagnosis is critical because effective treatments prevent recurrent seizures in “positive” cases, and in “negative” cases alternative (e.g. cardiac) interventions may be necessary. Nevertheless, most epilepsy diagnoses in the U.S. are made by non-specialists, and worldwide, most people with epilepsy lack access to diagnostic and treatment resources. There exists a critical need to increase access to reliable epilepsy diagnostic testing. Our group previously showed that a deep learning (DL) AI algorithm was able to identify epileptiform discharges (ED) in the EEG, the key physiologic biomarker of epilepsy, more accurately than individual epilepsy specialists. However, before such an AI can be used by non-specialists to accurately diagnose patients with suspected epilepsy, further work remains. Our central hypothesis is that point-of-care EEG technology, automated interpretation of EEG, and a simple questionnaire can enable non-experts to deliver accurate diagnostic services to patients with suspected epilepsy. The work for this project will be accomplished through three specific aims: SA1) To render interpretation of routine EEG accessible in underserved settings, we will develop a comprehensive AI approach to detecting epileptiform abnormalities. We will also train the new AI to classify the type of epilepsy based on the EEG. This aim will require obtaining annotations of 10K EEGs. These will also be made available as a resource to the research community. SA2) To allow non-experts to diagnose epilepsy as accurately as experts, we will develop a risk score that uses clinical history and AI-EEG to predict seizure recurrence. The score will be developed from prospectively collected data from 1000 patients. SA3) Aim 3: To validate and optimize our system for diagnosing epilepsy, we will test our point-of-care method in the target underserved patient groups in both a Boston-based Emergency Department as well as partner with health care workers in Guinea, an underserved area, to field-test the system in 600 new participants. This work will provide four key deliverables. 1) Ability to obtain EEGs and automated expert-level EEG interpretations in real-time at the point of care; 2) a unique, massive, public annotated dataset; 3) an epilepsy risk questionnaire that non-experts can administer and combine with automated EEG interpretation to make an accurate clinical diagnosis; 4) proof-of-concept for providing epilepsy diagnostic services in underserved areas.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT The long-term goal of this project is to understand the pathological mechanisms of sulfur mustard gas keratopathy (MGK) in the cornea. Sulfur mustard (SM) is an alkylating agent that has been used as a chemical warfare agent. SM exposure to the eye results in acute corneal injury. A subset of patients, particularly those with high exposure levels, develop chronic or delayed symptoms, which is known as MGK. Thus far, there are no specific treatments available to stop or reverse the detrimental effects of MGK. One of reasons for the lack of a specific treatment is that the mechanisms of MGK are not fully understood. Autophagy is a process by which cells break down and recycle their own cellular components, including damaged proteins and organelles. Even though autophagy has been recognized as a fundamental cellular process against stress, autophagy can play beneficial or detrimental roles depending on the context. In the cornea, it has been demonstrated that in response to most of stresses, autophagy plays beneficial roles to protect tissue homeostasis. Our laboratory and many other investigators have been focusing on such protective roles of autophagy in the cornea. However, the detrimental role of autophagy in the cornea has not been studied. Interestingly, when we investigated the role of autophagy in corneal injury due to chemical exposure, we found that nitrogen mustard (NM), an analog of sulfur mustard, induced a unique autophagy, which plays a harmful role in the cornea. It has been shown that the liberation of Beclin1, a key regulator in induction of autophagy, from Beclin1-Bcl2 complex can induce autophagy. Our preliminary data suggest that after NM exposure, sequestration of Beclin1 in Beclin1-Bcl2 complex attenuates NM-induced corneal inflammation. Therefore, we hypothesize that corneal mustard exposure induces autophagy via liberating Beclin1 from Beclin1-Bcl2 complex and such induced autophagy promotes corneal inflammation and contributes to MGK. In Aim 1, we will explore: (i) whether NM exposure will affect Beclin1-Bcl2 binding in vitro and in vivo; and (ii) whether manipulation of Beclin1-Bcl2 binding will affect NM-induced autophagy in cornea. In Aim 2, we will capitalize on our ability to conduct gain- and loss-of-function studies of induced autophagy in mice. We will inhibit induced autophagy via either reducing Beclin1 expression or preventing the disassociation of Beclin1-Bcl2 complex in vivo. We will also enhance induced autophagy via preventing the binding of Beclin1 to Bcl2 in vivo. We will utilize these genetically modified mouse models to determine whether the detrimental effects of NM exposure in cornea will be: (i) attenuated by inhibition of induced autophagy, while (ii) increased by enhancement of induced autophagy. Finally, in Aim3, we will test whether transient, pharmacological inhibition of autophagy attenuate NM-induced corneal injury. Knowledge from this project will reveal the pathological importance of induced autophagy in corneal MGK and will form the foundation for the development of novel therapies for this disease by targeting this Beclin1-Bcl2 complex-regulated autophagy pathway.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Electroconvulsive therapy (ECT) is a highly effective treatment for severe treatment-refractory depression and other conditions, in which carefully titrated electrical stimulation elicits brief, generalized seizures to change the brain to improve symptoms. An abundance of longitudinal MRI studies report robust and replicable brain plasticity after ECT, including increased hippocampal gray matter. However, it remains unclear how or why seizures are therapeutic in this context. Epilepsy research has demonstrated that seizure activity progresses through different brain regions and networks. Initial detection of seizure activity often occurs in a specific brain region (e.g., in medial temporal lobe), which can propagate locally, and in some cases spread via highly coordinated thalamo- cortical activity during generalization. Endogenous processes terminate the seizure, involving regions like anterior thalamus, basal ganglia, and cerebellum. A similar process appears to occur in ECT targeting temporal lobes, where electrical current initiates seizure activity in seizure-genic regions of medial temporal lobe (MTL), progressing to generalized seizure activity. Some seizure-network nodes have been implicated in antidepressant response to ECT, including parts of the hippocampus and thalamus. However, many seizure-network nodes are understudied in both ECT and epilepsy research in humans, because they are not included in standard MRI atlases (e.g., piriform cortex, substantia nigra, cerebellar nuclei) or due to limited spatial resolution in other neuroimaging techniques (e.g., coarse spatial resolution in molecular imaging, difficulty resolving deep structures in scalp EEG, limited number and position of pre-surgical recording electrodes in intracranial EEG). Thus, a precise, comprehensive understanding of seizure-network connectivity both in therapeutic seizure in ECT and pathological seizure in epilepsy remains elusive. The proposed studies will leverage pre-existing multi-modal MRI datasets to provide fundamental, mechanistic knowledge of entire seizure-network function before and after therapeutic and pathological seizure. The overall goal is to understand how seizure-network nodes interact in typical states and after therapeutic and pathological seizure, and to use that mechanistic knowledge to improve the administration of ECT, by using pre-treatment brain state to predict susceptibility and response to seizure therapy and by manipulating stimulus dose to influence the putative site of seizure initiation. Beyond improving the administration of ECT, the proposed studies have the potential to inform new neuromodulation strategies for depression, epilepsy, and other disorders, and to further knowledge of brain network function.

NIH Research Projects · FY 2026 · 2026-05

SUMMARY This research broadly seeks to reveal molecular principles that enable biological systems to sense and adapt to changing environments, and to understand how to use these principles to engineer synthetic biological systems that benefit humankind. A focal point of this work is RNA – a fundamental component of all living systems that enacts genetic, regulatory and catalytic functions by folding into intricate shapes within cells. As RNA folding can occur immediately during transcription, this raises a fundamental question as to how nascent RNA structures can influence many aspects of gene expression including transcription, translation, splicing, and polyadenylation. The overall vision for this proposed research program is to address aspects of this question through detailed structure-function studies of riboswitches and model eukaryotic systems, and development of technologies that can measure and model nascent RNA folding in broad RNA systems. Riboswitches are broadly distributed non- coding regulatory RNAs that bind to specific ligands and in response control gene expression. A critical feature of many riboswitches is that regulation occurs during transcription, making them ideal model systems to study the impacts of nascent RNA structure on gene expression. Principles of nascent RNA folding and function learned from riboswitches is beginning to be shown to hold true in other areas of biology including non-coding RNA biogenesis and eukaryotic RNA Pol III transcription termination. Our goal is to develop a molecular understanding of how cotranscriptional RNA folding regulates and coordinates cellular gene expression. Towards this goal, over the next five years we propose two research themes. The first uses diverse riboswitches, and eukaryotic RNA Pol III transcription termination, as model systems, to uncover general, quantitative relationships between RNA sequence, nascent folding, and transcription termination and translation initiation. We will combine high throughput cellular and in vitro functional characterization approaches to rapidly characterize model regulatory RNA sequence variants, with a hybrid experimental-computational approach called Reconstructing RNA Dynamics from Data (R2D2) to measure and model cotranscriptional RNA folding. Biophysical and data-driven modeling will be used to uncover quantitative relationships to address fundamental questions about how RNA folding kinetics and nascent RNA-RNA polymerase interactions govern transcription and translation regulation. Theme two will incorporate RNA folding energetics extracted from RNA structure chemical probing to improve R2D2’s accuracy and integrate new computational RNA folding algorithm developments to allow R2D2 to reconstruct RNA structure sub-populations of longer RNA systems. Detailed knowledge of how nascent RNA folding links to RNA regulatory function will contribute to a deeper understanding of gene expression processes, as well as numerous RNA biotechnologies such as diagnostics, RNA-targeting antibiotics and drugs, and nucleic acid therapeutics.

NIH Research Projects · FY 2026 · 2026-05

Project Summary: The overarching goal of this application is to investigate cell mechanics-mediated regulation of hair follicle stem cells (HFSCs) during homeostasis and aging. We propose to study microRNA-205- mediated regulation of extracellular matrix (ECM) and actin cytoskeleton for HFSC quiescence and activation and leverage the ability of microRNA-205 (miR-205) to stimulate HFSC activation to enhance HFSC aging. MicroRNA (miRNA) is a class of small noncoding, regulatory RNAs that play important roles in mammalian development, stem cells, diseases and aging. In our preliminary studies, we have determined mechanical properties of HFSCs during homeostasis and aging. We have revealed that bulge HFSCs reside in a stiff microenvironment with high actomyosin contraction forces. In contrast, hair germ progenitors are relatively soft and undergo periodic enlargement and contraction. Notably, induction of miR-205, one of the most highly expressed miRNAs in HFSCs, downregulates many bona fide targets, which are enriched in the function of ECM, actomyosin cytoskeleton and mechanosensing. And this leads to rapid activation of HFSC cell division and promotes hair regeneration in both young and aged mice. Mechanistically, we have identified Piezo1 as a novel target of miR-205, which functions downstream of miR-205 and translates mechanical cues into a gene expression program to reinforce the mechanical properties and maintain cellular states of quiescent HFSCs. To examine the role of PIEZO1-mediated calcium influx in HFSCs, we have further developed a high-resolution intravital imaging system to accurately record calcium influx in HFSCs over an extended period of time during quiescence and activation. This allows us to quantify cumulative calcium levels and further identify transcription factors, NFATC1 and JUN (AP1), which function downstream of PIEZO1-mediated calcium influx to promote the expression of the ECM and actin cytoskeleton genes. Based on these exciting findings and promising preliminary data, we propose to further elucidate the mechanism of miR-205-mediated HFSC activation and aging through the regulation of ECM and actomyosin contraction forces (Aim 1), determine the regulation of PIEZO1-mediated mechanosensing by miR-205 (Aim 2), and leverage miR-205-induced HFSC activation to improve HFSC functions and hair growth during aging (Aim 3). Together, this application will provide new insights into the mechanisms orchestrating the mechanical properties and stem cell functions of HFSCs. By harnessing the powerful combination of live imaging, cell biology, mouse genetics, and single-cell genomics, we will establish a new paradigm for studying tissue architecture, cell mechanics and underlying mechanisms. These results will lay the foundation for leveraging noncoding, regulatory RNAs to enhance HFSC functions during aging.