Northwestern University

NIH Research Projects · FY 2026 · 2026-02

Project Summary/Abstract Human papillomaviruses (HPV) infect stratified epithelia and link their productive life cycles to differentiation. The HPV life cycle is regulated by viral proteins acting together with cellular factors that control cell cycle progression, differentiation, and DNA damage repair. This application will investigate the pathways that regulate the productive replication of high-risk papillomaviruses. We have previously shown that activation of both the ataxia telangiectasia (ATM) pathway as well as the ataxia telangiectasia and Rad3-related (ATR) DNA damage repair (DDR) pathway is necessary for differentiation-dependent amplification of viral genomes. In addition, ATR activation was found to play a critical role in the stable maintenance of episomes in undifferentiated cells. Our studies further indicate these pathways are activated by the high numbers of DNA breaks caused by aberrant R-loops. R-loops are trimeric nucleic acid structures consisting of a hybrid between RNA and its complementary DNA strand along with the displaced single strand DNA. These are stable structures that form at promoter as well as termination regions. R-loops play important roles in the normal regulation of transcription initiation and termination; however, failure to resolve aberrant R-loops leads to DNA break formation. We have shown that R-loop levels are increased by over 10-fold in HPV positive cells, and maintenance of these high levels is necessary for viral replication and transcription. Furthermore, our work indicates that these enhanced R-loop levels help to regulate the expression of important cellular pathways, including the repression of innate immune regulatory genes. Two major enzymes act to regulate R-loops; RNAse H1 is an R-loop specific RNase that degrades the RNA moiety while senataxin (SETX) is a helicase that unwinds R-loops as well as acts at termination sites that contain R-loops to recruit the exonuclease XRN2 for mRNA cleavage. Our recent work shows that both SETX and RNAse H1 contribute to repression of innate immune gene expression in HPV positive cells while at the same time both are required for high level viral transcription. SETX acts to regulate resolution of R-loops particularly those at 3’ termination sequences along with activating the m6A RNA methylation pathway. Recruitment of the m6A catalytic enzyme Mettl3 is mediated by SETX to methylate RNAs in R-loops and is important for their resolution and proper termination of transcription. RNAse H1 acts primarily to remove RNA moieties in R-loops. These two enzymes appear to work through complementary but different mechanisms and this application proposes to investigate how this leads to repression of innate immune gene expression along with activation of viral expression and replication.

NIH Research Projects · FY 2026 · 2026-02

Macrophages (MFs) play many critical roles in tissue health and disease pathologies. Tissue-resident MFs maintain tissue homeostasis and fulfill key functions, while monocyte-derived MFs infiltrate the tissue upon challenge and drive the inflammatory responses. Autoimmune diseases, such as Rheumatoid Arthritis (RA), are characterized by recurring inflammatory flares driven by MFs with an impaired transition to resolution and wound healing that leads to exacerbated tissue damage and worsened disease outcomes. Such damaging inflammation may be modulated by trained immunity, whereby prior stimulation of MFs leads to a functional memory that drives response to future immune challenges. Thus, approaches to train MF functional memory by limiting their inflammatory response and elevating their pro-repair functions is critical to the development of therapeutic strategies to maintain remission states by preventing disease flares. Our overarching goal is to investigate neutrophil (PMN) priming as a method of MF functional training. Emerging evidence demonstrates that PMNs in acute inflammation can instruct MF activity as a part of normal immune response. This includes the well- established MF reprogramming during the efferocytosis process in the disease resolution phase, as well as via less understood contact or soluble factors dependent interactions. However, whether such training also leads to innate immune MF memory is unknown. Our preliminary data demonstrate frequent dynamic interactions of live PMNs with tissue resident/infiltrating MFs in inflamed tissue in several inflammation models, indicative of an active crosstalk and potential cross cell instruction. Furthermore, studying PMN-MF interactions through transwells revealed that ex vivo pro- vs anti-inflammatory PMN stimulation/priming differentially impacted MF transcriptional regulation without phagocytosis. Thus, we hypothesize that primed PMNs can trigger MF trained immunity, promoting their pro-repair activity in inflamed joint. We will test this hypothesis by determining whether PMNs can train MF functional memory guiding long-term responses to inflammatory stimuli and whether this can help resolve inflammation in vivo. In Aim1, we will establish the impact of differentially primed PMNs on MF functional memory using transwell co-cultures to assess the impact of pro- vs anti- inflammatory priming of PMN on MF responses and their functional memory upon inflammatory rechallenge. In Aim 2, we will determine if PMNs instruction of MFs can be used to attenuate and/or resolve inflammation in STIA (serum-transfer-induced arthritis), a mouse model of RA, by adoptively transferring primed PMNs into mouse joints prior to disease onset. While memory is a well-studied feature of adoptive immunity, its potential for innate immune cells has not been fully explored. Thus, through these conceptually and technically innovative experiments, we expect to establish the possibility of long-term innate immune instructional crosstalk resulting in functional memory and identify novel regulatory mechanisms of trained immunity in MFs with important implications for designing future treatments of inflammatory disorders.

NIH Research Projects · FY 2026 · 2026-02

Summary Viruses require a variety of host cell proteins to enable them to replicate, and identification of host factors that contribute to virus replication, fitness, and tropism is a top priority for basic and applied research. Zika virus (ZIKV) is a Flavivirus known to cause significant human disease and mortality and can cross the placental barrier to infect unborn children. Despite its direct impact on human health there are currently limited therapeutic options available. It is essential therefore to identify the factors that contribute to ZIKV infection and understand their modes of action. Cellular ubiquitin (Ub) systems are important for regulating fundamental eukaryotic processes and mediating both innate and adaptive immunity. Ub pathways are often appropriated by viruses to directly or indirectly influence their replication. We used an original live cell-based high-throughput screen was used to identify Ub-related genes that are required for ZIKV replication. Rigorous primary, secondary, and counter screening was used to identify host factors that impact ZIKV replication. Based on the intensity and reproducibility of its impact on ZIKV replication, TRIM39, a Ub ligase not previously connected to ZIKV, was advanced as a promising target for further investigation. Additional results demonstrate that TRIM39 disruption restricts both 1947 and 2015 ZIKV isolates, decreases infectious virus production by ³100-fold, limits ZIKV RNA and protein production, and reduces antiviral responses. These properties suggest that TRIM39 and its Ub substrates have high potential for development as anti-ZIKV therapeutic targets. However, little is known about TRIM39 itself or how it may interface with ZIKV. Experiments indicate TRIM39 level is decreased during ZIKV infection, and that it is specifically antagonized by the ZIKV NS5 protein, a multifunctional viral protein responsible for RNA synthesis and innate immune evasion, leading to proteasomal degradation. We find that NS5 engages the TRIM39 SPRY domain in a complex that leads to this viral hijack, suppression, or antagonism. As a previously unrecognized host protein that can modulate ZIKV infection, we postulate that TRIM39 is needed in the positive progression of ZIKV infection and may target inhibitory or antiviral factors that otherwise would hinder ZIKV replication. The ZIKV NS5 protein modulates TRIM39 actions via a protein interaction and degradation. Three Aims will test these hypotheses, reinforce TRIM39’s significance for ZIKV and other RNA viruses, and reveal its roles in ZIKV infections. We will (i) expand analysis of TRIM39 for ZIKV in placental trophoblast derived host cells, and for both closely related (Dengue) and unrelated (IAV, VSV) RNA virus infections, (ii) identify TRIM39 interaction partners and Ub substrates in uninfected and infected cells, and (iii) determine the mechanistic basis of NS5-TRIM39 interaction and its impact on the ZIKV life cycle. These goals will produce a more complete understanding of TRIM39 in ZIKV biology, reveal the mechanisms underlying its essential role in infections, and authenticate a new mediator of virus infection.

NIH Research Projects · FY 2026 · 2026-02

Project Summary/Abstract Store-operated Ca2+ release-activated Ca2+ (CRAC) channels play a central role in cellular Ca2+ signaling across diverse cell types including T-cells, microglia, and astrocytes. Activated by the endoplasmic reticulum (ER) Ca2+ sensor, STIM1, CRAC channels regulate a wide variety of effector cellular functions including transcription, metabolism, and cell proliferation. Their unique biophysical properties, characterized by exceptional Ca2+-selectivity, store-operated activation, and distinct gating modes, make them fascinating models for understanding the mechanisms of Ca2+ permeation and gating. Moreover, because CRAC channels sit squarely at the center of cellular Ca2+ signaling networks in many cells, aberrant CRAC channel function is implicated in the etiology of diseases ranging from immunodeficiency to allergies. Previous work on the molecular choreography of the CRAC channel activation process has revealed that opening of CRAC channels is mediated by direct interactions between the pore-forming Orai proteins with STIM1. Despite progress in elucidating the activation process of CRAC channels, numerous key aspects of the molecular and structural mechanisms of STIM1-Orai1 binding, channel pore opening, and regulation of gating remain unknown. Here, we propose a multi-disciplinary approach that will integrate photocrosslinking of Orai1 with the catalytic domain of STIM1 using unnatural amino acids, structural analysis of open Orai channel variants using cryo-electron microscopy, and use of fluorinated phenylalanine analogs to elucidate the molecular and structural basis of STIM1-Orai1 binding and the gating of Orai1 channels. Our goals are to: (1) determine the molecular and structural basis of the Orai1-STIM1 binding interface, (2) determine the structures of three gain-of function Orai mutants (including a human disease mutation) with distinct biophysical and functional properties via Cryo-electron microscopy and compare these to the closed, wildtype channels, and, (3) examine the contributions of electrostatic interactions involving the channel gate that mediate the opening of the pore. These studies will significantly advance our understanding of the molecular and structural mechanisms of CRAC channel gating, reveal how disease mutations alter these processes, and ultimately provide new therapeutic strategies for targeting CRAC channels in disease.

NIH Research Projects · FY 2025 · 2026-02

PROJECT SUMMARY. Neurodegenerative diseases are currently defined by the clinical symptoms observed antemortem (i.e., dementia syndrome), patterns of anatomic atrophy, and cellular and molecular pathology confirmed postmortem. A key challenge in this field is the fact that a single dementia syndrome can be caused by multiple pathologies, and the same pathology can manifest as multiple dementia syndromes. The “Type C” variant of frontotemporal lobar degeneration due to TDP (TDP-C) pathology is unique. TDP-C shows a strong affinity for the anterior temporal lobe (ATL), a brain region critical for word knowledge. TDP-C also has an uncommonly tight coupling with semantic primary progressive aphasia (PPA-S), a dementia syndrome defined by initial decline primarily in semantic (i.e., word knowledge) abilities and atrophy of the ATL in the left language hemisphere; nearly 90% of PPA-S cases show TDP-C at death. The association of selective ATL atrophy, TDP- C pathology, and semantic language impairment in PPA-S offers an ideal paradigm to examine anatomic language networks, mechanisms of neurotoxicity, and principles of selective vulnerability. A central hypothesis is that atrophy patterns in PPA-S mirror underlying neurodegenerative changes due to TDP-C, leading to dysfunction in neurocognitive networks critical for semantics. The proposed research will characterize and clarify relationships between clinical symptoms, atrophy, and underlying pathological substrates in PPA-S due to TDP- C. Aim 1 of this study focuses on identifying the anatomic signature unique to semantic language impairment progression seen in PPA-S due to TDP-C. It is hypothesized that this progression is defined by progressive atrophy along the temporal lobe of the left language hemisphere. Longitudinal neuropsychological testing will categorize TDP-C individuals into discrete stages of semantic language impairment. Volume loss will be assessed using voxel-based morphometry, comparing whole brain volume and specific regions of interest (ROIs) along the temporal lobe in TDP-C participants to a cognitively healthy group at and between impairment stages. The goal of Aim 2 is to examine the clinicopathologic relationship between antemortem patterns of atrophy and postmortem pathologic markers in the temporal lobe in PPA-S. It is expected that regional and hemispheric densities of markers will mirror the aphasic phenotype and atrophy pattern in PPA-S. Using whole hemisphere sections, pathologic inclusions, neuronal and synaptic integrity, and neuroinflammatory (microglia) markers will be quantified in bilateral ROIs used in Aim 1 using unbiased stereology and digital pathology methods. This project will occur within the Northwestern University Alzheimer’s Disease Research Center, a multidisciplinary, international PPA referral center that holds the world’s largest cohort of TDP-C participants with extensive longitudinal clinical, neuroimaging, and neuropathologic data. The public health impact of this study is significant, as studies of TDP-C have the potential to address questions about dysfunction due to neurodegenerative pathology and inform future development of disease-specific biomarkers and therapeutic targets.

NIH Research Projects · FY 2026 · 2026-02

PROJECT SUMMARY The sense of taste informs the nutritional value and safety of potential food sources. Because this assessment is critical for the survival of the animal, taste signals immediately evoke stereotypical behaviors (e.g., licking, salivating, retching) leading to acceptance or rejection of the food item. Thus, the taste sensory system presents an exquisite neurobiological circuit to investigate how the brain accurately processes the external environment and sets into motion hardwired behaviors. The overall objective in this application is to understand how taste quality (sweet, bitter, umami, salty, sour), intensity (concentration), and valence (attractive, aversive) information is encoded by the peripheral gustatory system – from the taste buds to gustatory ganglia – before it gets processed in the brain. The central hypothesis posits that while TRCs and gustatory neurons may broaden their tuning, certain taste qualities (particularly those encoding different valences – appetition vs aversion) should remain reliably distinct. This hypothesis will be tested in mice by addressing the following three specific aims: 1) Assess tastant-evoked activity in TRCs in vivo; 2) Visualize tastant-evoked responses in intragemmal gustatory nerve fibers; and 3) Interrogate synaptic-level connectivity in the taste bud. The proposed research is significant because it will provide new insights on how taste information is coded at the periphery.

NIH Research Projects · FY 2026 · 2026-01

SUMMARY Death induced by Survival Elimination (DISE) is a powerful cell death mechanism in which a class of short RNAs (sRNA) specifically targets a network of genes critical for cell survival in a miRNA-like fashion. G-rich 6mer containing sRNAs kill by targeting C-rich seed matches enriched in the 3'UTR of survival genes (SGs). The kill code is found in certain tumor suppressive miRNAs and it developed over at least 800 M years. Interestingly, we noticed that in tumor bearing animals treated with these toxic sRNAs, while tumors slowed down in growth, normal tissue were not affected. We proposed that at least four different mechanism ensure that normal tissues are protected from DISE. These include global downregulation of miRNAs in all cancers compared to normal tissue, most of which carry nontoxic AU-rich seeds and an upregulation of the targeted SGs in all cancers (new preliminary data in this proposal). We wondered what would happen if the DISE mechanism became overactive and/or the protection against it was lost. Would this result in pathology in affected tissues? With this important question in mind we ventured into neurodegenerative diseases. We found evidence for toxic sRNAs to contribute to Huntington’s disease (HD). Another interesting link we found was with aging during which many tissues including the brain lose expression of abundant nontoxic miRNAs in part we posit to protect themself from endogenous toxic sRNAs. This resulted in a fundamental hypothesis that many neurodegenerative diseases have a toxic RNA component and that a gradual age related loss of protective miRNAs in the brain is one reason that these diseases often have decades of symptom free live before the onset of symptoms. This led to a recent study in which we provided first evidence that DISE contributes to Alzheimer's disease (AD). Consistent with our hypothesis for major neurodegenerative diseases it has been reported that when pathology occurs cancer incidence rates go down consistent with an overactivity of the anti-cancer mechanism. This was shown for HD and Spinal-Bulbar Muscular Atrophy (SBMA), AD, Parkinson's disease and Amyotrophic Lateral Sclerosis (ALS). Based on these data we propose this central hypothesis: Normal tissues are protected from DISE by high amounts of protective miRNAs, expression of which gradually declines during aging making tissues less resilient to a number of stressors causing tissue degeneration. Cancer and many degenerative diseases are therefore two sides of the same coin and in order to understand and eventually treat either of these major diseases the two sides and how they affect each other have to be better understood. In this proposal we will study fundamental mechanisms that allow certain cells to be eliminated while others being protected from DISE and assess what happens when that protection is lost either during aging or by eliminating protective miRNAs. This will be tested in four specific aims: Aim 1: Determine whether the kill code is universal or specific for different cancers and their originating tissues. Aim 2: Identify mechanisms that protect normal cells from DISE. Aim 3: Determine the effects of modulating DISE in AD. Aim 4: Explore changes in DISE sensitivity during aging.

NIH Research Projects · FY 2025 · 2026-01

PROJECT SUMMARY The Neisseria gonorrhoeae (the gonococcus or Gc) Type IV pilus (T4p) is a critical colonization and virulence factor during pathogenesis. T4p are used to adhere to host cells and promote colonization. T4p are dynamic structures made of pilin fibers that extend and retract from the bacterial cell. Beyond host cell adhesion, T4p are involved in a variety of cellular processes including DNA uptake, twitching motility, and resistance to antimicrobial agents. We previously identified a novel component of the Gc T4p, TfpC. Our published data illustrated that TfpC is required for full piliation, and piliation is restored in tfpC mutants lacking the PilT retraction ATPase. There are eleven other such mutants in the Gc T4p. Here, we use genetic, biochemical, and biophysical techniques to define how TfpC contributes to T4p dynamics and architecture. We show that while TfpC is required for complete piliation of Gc, remaining pilus structures undergo extension at rates similar to parental strains. However, retraction is slower in strains lacking tfpC. Furthermore, we demonstrate that TfpC stability is dependent upon the PilQ secretin. We also identify an important motif within the flexible, disordered, proline-rich region of TfpC that is important for function or stability of TfpC. I hypothesize that TfpC interacts with the PilQ secretin or other components of the T4p to support a subset of T4p. To test this hypothesis, I will 1) determine if TfpC interacts with the PilQ secretin, 2) perform structure-function analyses of TfpC, and 3) interrogating the mechanism underlying reduced piliation in twelve pilus mutants. The proposed research is significant because it will define the previously uncharacterized TfpC protein required for piliation and link its activity to other pilus components. It will also provide important information for other species T4p systems and possibly some Type II secretion systems. These studies expand our current understanding of T4p architecture as well as the components governing mechanisms of extension and retraction, providing a foundation for future studies on the T4p dynamics essential for colonization and virulence. Long-term, the proposed research could provide new targets for treating gonorrhea infections.

NIH Research Projects · FY 2025 · 2025-12

PROJECT SUMMARY/ABSTRACT The SCN2A gene, which encodes a neuronal voltage-gated sodium channel, is a major genetic risk factor for a wide range of neurodevelopmental disorders, including childhood epilepsies, autism spectrum disorder, and intellectual disability. SCN2A channels are primarily expressed in axon initial segments and dendrites of excitatory glutamatergic neurons, where they drive action potential initiation and backpropagation, respectively. Pathogenic SCN2A mutations affecting protein structure and function modulate neuronal responses to synaptic inputs leading to the cognitive and behavioral impairments and seizures observed in SCN2A-related disorders. We discovered that the location of variants within the SCN2A protein is strong predictor of channel dysfunction, accounting for approximately 40% of the variance in channel biophysical properties. However, the net effects of complex and/or opposing channel biophysical properties on excitatory neuron excitability are difficult to predict and the relationship between specific channel biophysical perturbations and resulting changes to neuronal physiology is unclear. In this project, we propose to determine the consequences of 115 disease-associated SCN2A mutations on channel biophysical properties and intrinsic physiological properties of neocortical pyramidal neurons (Aim 1A). To do this, we will leverage in vitro whole-cell voltage-clamp recordings to build in silico hidden Markov models of each SCN2A variant, which we will use to simulate heterozygous channel dysfunction in a benchmarked mathematical model of a neocortical pyramidal neuron. Subsequently, we will use interpretable machine learning algorithms to explain the relationship between functional perturbations and neuronal physiology (Aim 1B). We also propose to validate neuronal simulations for 10 SCN2A mutations using an experimental system called dynamic action potential clamp (Aim 2A). In this system, we will electrotonically couple mammalian cells heterologously expressing SCN2A variants to a benchmarked model of an axon initial segment and determine effects of heterozygous channel dysfunction on simulated neuronal excitability. Finally, we aim to study 2 prototypical gain-of-function SCN2A mutations that heighten channel activity, potentiate pyramidal neuron firing, and lead to seizures, and evaluate the feasibility of using dynamic action potential clamp to test functional correction of abnormal neuronal excitability using specific sodium channel blockers (Aim 2B). The long-term goal of this entire project is to computationally map and experimentally validate the relationships among protein structure, channel dysfunction, neuronal physiology, and clinical phenotypes in SCN2A-related disorders. Overall, our work will generate a granular genotype-phenotype relationship landscape of SCN2A- related disorders revealing underlying molecular and cellular mechanisms of pathophysiology and will advance a new approach methodology for testing precision medicine interventions for treating neurological disorders.

NIH Research Projects · FY 2026 · 2025-12

PROJECT SUMMARY Haploinsufficiency of chromatin remodeler CHD2 causes a neurodevelopmental disorder (NDD) characterized by developmental delay, intellectual disability, and epilepsy. Adjacent and upstream of CHD2 is a conserved long non-coding RNA (lncRNA) CHASERR. Deletion of CHASERR causes CHD2 overexpression and a more severe, early onset developmental disorder with significant motor and language delay, intellectual disability, and structural brain defects in humans. RNA-seq and western blot analysis of patient-specific induced pluripotent stem cells (iPSCs) and CRISPR-generated HAP1 cells have shown that CHASERR deletion increases CHD2 expression and protein levels in cis. While there is growing evidence of the role of lncRNAs in gene regulation, the mechanism of how CHASERR regulates CHD2, and the downstream consequences of too much CHD2 on global chromatin dynamics and neurodevelopment, is not well understood. Prior studies suggest that CHASERR is concentrated within its locus and binds to SPEN, a protein known to recruit HDAC3 and other chromatin remodeling proteins to repress transcription. Cleavage Under Targets and Release Using Nuclease (CUT&RUN) in HAP1s showed loss of HDAC3 occupancy at CHD2 locus in CHASERR knockout (KO) but not wildtype (WT), suggesting that the CHASERR-SPEN complex is essential to recruit HDAC3 and repress CHD2 expression. Taken together, I hypothesize that CHASERR deletion results in loss of recruitment of SPEN and other repressive proteins at the CHD2 locus, leading to a more open chromatin state permissive of increased CHD2 expression, resulting in CHD2 overproduction and global changes in chromatin dynamics. Using CRISPR-generated CHASERR KO and antisense oligonucleotide (ASO) to knockdown CHASERR in WT HAP1 cells, as well as patient-specific iPSCs and neural progenitor cells (NPCs), I will address my hypothesis with two aims. In Aim 1, I will conduct CUT&RUN on chromatin remodeling proteins (HDAC3, CHD2, EZH2) and histone modifications (H3K27ac, H3K27me3) to determine which proteins CHASERR and SPEN recruit to change local chromatin structure and repress CHD2 expression. In Aim 2, I will use a multi- omics approach with WT, ASO knockdown, and patient-derived CHASERR+/- NPCs, to understand how CHD2 overexpression affects global chromatin dynamics and transcription in a neural model. Because treatment for CHD2-related NDDs requires precise dosage control of functional CHD2, the proposed studies will help understand CHASERR’s regulation of CHD2 and its potential as a therapeutic target for CHD2 patients. Furthermore, this study will contribute to broader understandings of lncRNA biology and the biological underpinnings of childhood developmental disorders. The diverse team of mentors and the premier facilities and equipment at Northwestern will be available throughout the award period to support rigorous training to carve a successful physician-scientist career uncovering genetic mechanisms of pediatric neurological disorders.

NIH Research Projects · FY 2025 · 2025-12

Project Summary/Abstract Successful communication is dependent on a multitude of intricately coordinated skills that facilitate connections between interlocutors, including prosody, or the rhythm, rate, and intonation of speech. Speech rhythm, specifically, is critical for communicating intent and enhancing speech comprehension through predictable, regularly-timed beats. Without rhythm, speech lacks the temporal guideposts that instruct the listener what to listen for and when, leading to miscommunications (i.e., “let’s eat, mom!” vs. “let’s eat mom!”). Amongst autistic individuals and their first-degree relatives, prosody, and speech rhythm specifically, has been identified as a salient contributor to social communication differences. Parallel findings have been reported in fragile X syndrome (FXS; the leading monogenetic cause of autism) and in FMR1 premutation (PM) carriers, who show similar but less striking rhythmic variability. Together, these findings suggest a shared, heritable, neurobiological mechanism underpinning speech and higher-order language skills impacted in autism, and linked to a known autism risk gene. Importantly, recent acoustic research has revealed links between rhythmic variability, speech rate, and FMR1-mediated molecular-genetic variation in females with FXS, who are largely understudied in FXS research and tend to show less cognitive impairment than males. Notably, recent work in autism has pointed to differences in cortical processing of continuous speech, whereas studies in FXS and the PM have identified articulatory abnormalities, as potential perceptual and speech-motor mediators of speech-rhythm differences, but their associations with speech rhythm remain largely unexplored. As such, evaluating the perception and production of continuous speech in relation to speech rhythmicity in FMR1-mediated conditions is a critical next step towards understanding the role of FMR1 in autism-related phenotypes. Further, exploring speech rhythm in females with FXS and the PM will allow for greater characterization of the neural and speech-motor mechanisms contributing to a broad spectrum of FMR1-mediated speech and language profiles, from clinically to sub-clinically affected, without the confounding impact of intellectual impairment. Given mounting evidence of impacted cortical speech processing and speech-motor differences in autism, as well as speech-motor incoordination in FMR1 conditions, we hypothesize that speech rhythmicity may be disrupted across in females with FXS and the PM and contribute to pragmatic language differences. Moreover, we hypothesize that differences in speech rhythm are differentially mediated by domain-general rhythmicity, speech-motor coordination, cortical speech perception, and rooted in underlying variation in FMR1-mediated molecular-genetic correlates. If completed, the proposed study has potential to provide key insights into the role of a known genetic mutation in the autism language phenotype with relevance to future behavioral interventions.

NIH Research Projects · FY 2025 · 2025-12

PROJECT SUMMARY Streptococcus pneumoniae is a significant cause of morbidity and mortality in the United States and around the world. There are over 100 known serotypes of S. pneumoniae, and conjugate vaccines are an effective tool to protect the public from infection of covered serotypes. These vaccines work by conjugating capsular polysaccharide from unique S. pneumoniae serotypes to an immunogenic carrier protein, which induces a T cell-dependent memory immune response. However, serotype replacement, a process in which serotypes that are not covered by a vaccine increase in incidence due to the removal of competing serotypes, necessitates higher valency vaccines. The conventional, chemical technology used to synthesize pneumococcal conjugate vaccines has substantial limitations that prevent the synthesis of higher valency vaccines. This technology randomly conjugates glycan to carrier protein, which destroys T cell epitopes on the protein, reducing immunogenicity of the vaccine. Increasing valency using this conventional strategy also decreases immune responses against each serotype due to carrier suppression, or the decrease in anti-glycan immune response with higher amounts of carrier protein included per dose of vaccine. Therefore, there is a need to develop novel and more highly immunogenic vaccines with enhanced immunogenicity to increase vaccine valency and reduce carrier suppression. Protein glycan coupling technology is a novel method to enzymatically conjugate a carrier protein with pathogen glycan using an oligosaccharyltransferase. This technology enables site-specific glycosylation of carrier protein to avoid T cell epitope destruction. The central hypothesis of this proposal is that site-specific glycosylation of carrier protein using enzymatic conjugation can improve the immunogenicity of pneumococcal conjugate vaccines. Recent advances to protein glycan coupling technology have enabled its use in cell-free systems, which allows for the high-throughput synthesis of hundreds of unique glycoconjugate vaccine designs. This project will evaluate three distinct features – glycan location, density, and delivery – for synthesis of glycoconjugates via site-specific enzymatic glycosylation (Aim 1) and evaluate the immunogenicity of top candidates in mice (Aim 2). In Aim 1, glycosylation at every potential location throughout CRM197, an FDA-approved carrier protein, will be evaluated in high-throughput using cell-free systems. Sites away from T cell epitopes that can be glycosylated will be iteratively combined to increase glycan density, and glycoproteins will be attached to synthetic vesicles to improve antigen presentation. In Aim 2, multi-dose vaccine trials will be performed in mice with top vaccine candidates from Aim 1 and compared to a conventional vaccine. The immunogenicity of each vaccine will be evaluated by measuring antibody titers, opsonophagocytic activity, memory B cell production, and mouse survival after S. pneumoniae challenge. The results will inform conjugate vaccine design for S. pneumoniae and could be applied to develop novel vaccines against other pathogens.

NIH Research Projects · FY 2025 · 2025-12

Project Summary/Abstract Understanding how a protein’s amino acid sequence controls biophysical properties like stability, folding, and reactivity is key to designing better enzymes. However, this task remains challenging due to the complexity of protein conformational ensembles and the scarcity of functional data across sequence space. As a result, computational enzyme design is difficult, and most designed enzymes fail to function without clear indications why. Large-scale experiments can help uncover how specific residues and higher-order interactions affect function, offering the basis for improved computational models and better strategies for enzyme design. However, most surveys of sequence space either cover relatively few variants, or sample only a very narrow region of sequence space deeply. Combining high-throughput functional mapping with de novo design of new enzymes has been out of reach due to the size and complexity of enzymes typically targeted in such experiments. To this end, we propose to map the sequence-function landscape of SpyCatcher, a small reactive protein that forms a covalent bond with its substrate SpyTag, investigating both natural and de novo sequences. While not a true enzyme, SpyCatcher’s small size (80 residues) and simple reaction mechanism make it a tractable minimal model for enzyme-like reactivity. In Aim 1, we will use cDNA display-based to measure the reactivity, substrate binding kinetics, and stability of ~ 1 million natural SpyCatcher variants. The unprecedented scale of these functional measurements on a uniquely simple system will allow us to rigorously explore additive and non-additive sequence-function relationships and identify predictive patterns using machine-learning models. In Aim 2, we will leverage de novo design to explore further reaches of sequence space and test different design strategies in high-throughput, creating SpyCatcher-like proteins with sequences far from natural ones. These designs (three rounds of ~500,000 sequences) will be assayed with the same high-throughput assays to iteratively inform our design process and improve criteria for designing proteins with functional reactivity. We aim to generate high-quality, large-scale datasets spanning both natural and unnatural sequence space in order to uncover fundamental "rules" of reactive protein design, which can inform future engineering efforts. By producing interpretable data that links sequence to biophysical properties, we aim to enhance our ability to design proteins with tailored functions.

NSF Awards · FY 2025 · 2025-11

Nanotechnology has rapidly emerged as an interdisciplinary field with transformative applications across medicine, energy, and national defense. At Northwestern University, the International Institute for Nanotechnology (IIN) leads one of the nation’s foremost research efforts in this area. The Research Experiences for Undergraduates (REU) Site in Nanoscale Science and Engineering at Northwestern will provide undergraduate cohorts from various science and engineering backgrounds with immersive, nine-week summer research experiences. Students will engage in hands-on, goal-driven projects under the mentorship of faculty, postdoctoral fellows, and graduate students, gaining critical thinking skills and exposure to real-world laboratory environments. An educated and motivated scientific workforce is essential to sustaining national progress in science and technology, and undergraduate research experience plays a pivotal role in cultivating this talent. This REU Site promotes interdisciplinary learning across chemistry, physics, biology, and engineering, preparing students to contribute meaningfully to the field of nanotechnology. Over the past decade, REU sites hosted by the IIN have trained more than 330 undergraduates and generated over 130 publications with student co-authors. This REU site is committed to broadening participation among students with limited access to research opportunities, with the goal of expanding interest in STEM and cultivating the next generation of scientists and engineers in the United States. There is a critical need for the creation of a strong workforce of world-class scientists, engineers, and educators with knowledge and technical skills in nanotechnology. Researchers at Northwestern University (NU) have made substantial contributions to this field, and the university has gained a reputation for its rigorous nanotechnology research programs. This nine-week, immersive summer program is designed to foster interdisciplinary collaboration, provide undergraduates with hands-on research opportunities in chemistry, biology, medicine, physics, and multiple engineering disciplines, all united by a focus on nanoscience and nanotechnology. The program’s recruitment and selection process will encompass inclusive practices to ensure that all outreach and participation opportunities are open and available to all Americans, in alignment with NSF’s commitment to broad accessibility. Each selected student will be carefully paired with a research project based on their noted interests and experience provided in their application. They will be guided by a faculty mentor and a graduate or postdoctoral researcher, with the goal of progressing from structured supervision towards independent experimentation. Participants gain experience in experimental design, data analysis, and scientific communication, supported by workshops in public speaking, technical writing, and research ethics. Additional activities include visits to a National Laboratory, seminars, and social events that foster community and professional development. Students present their research at a final symposium and in a written technical report, which may be submitted for publication. Through this program, participants acquire the skills and experience necessary for future success in furthering their education, and/or entering the American workforce, securing our nation’s leadership at the forefront of science and technology. 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 2025 · 2025-11

PROJECT SUMMARY The rising prevalence of food allergies underscores the urgent need to unravel the pathogenesis of this disease and to develop effective therapies. Currently, our knowledge of the biological rules that govern which individuals respond to food allergens is incomplete. While food-specific IgE is necessary for symptomatic food allergy, it is not sufficient; some individuals with food specific IgE exhibit no allergic reaction upon eating foods they are sensitized to. This suggests that additional factors beyond IgE contribute to anaphylaxis risk. Our lab has demonstrated that, unlike C3H/HeJ and BALB/c mice, C57BL/6 mice exhibit minimal allergen absorption across the gut and are resistant to developing an anaphylactic response following oral challenge (termed “oral anaphylaxis”). Our forward genetic screen of oral anaphylaxis-susceptible and resistant mice identified allelic variants in the gene Dipeptidase 1 (Dpep1), which encodes an enzyme in the cysteinyl leukotriene synthesis pathway, as a key predictor of anaphylactic response. Further, we found that blocking leukotriene synthesis entirely with the drug zileuton significantly reduced gut permeability and protected sensitized, oral anaphylaxis- susceptible C3H/HeJ and BALB/c mice from anaphylaxis following allergen challenge. My proposal therefore investigates this novel role for cysteinyl leukotrienes as mediators of food allergen uptake across the gut barrier. My central hypothesis is that intestinal mast cell production of the cysteinyl leukotrienes LTC4 and LTD4 and their signaling via the cysteinyl leukotriene receptor 2 mediate gut absorption of food allergens and dysregulation of this pathway can result in enhanced risk of anaphylaxis after food sensitization. In my first aim, I will identify the specific cysteinyl leukotriene(s), cysteinyl leukotriene receptor(s), and cellular producers of cysteinyl leukotrienes relevant to absorption of allergens in the gut. To accomplish this, I will assess the effects of exogenous cysteinyl leukotriene administration, cysteinyl leukotriene receptor knockouts, and mast cell depletion on gut permeability and susceptibility to oral anaphylaxis. In my second aim, I will determine whether aspirin, a known clinical cofactor of anaphylaxis, enhances gut permeability and susceptibility to anaphylactic response by increasing cysteinyl leukotriene levels. To do so, I will evaluate the effects of aspirin treatment on cysteinyl leukotriene levels in the small intestine and intestinal absorption of food allergens in sensitized mice. Next, I will test whether cysteinyl leukotriene receptor knockout mice or pretreatment with leukotriene synthesis inhibitors decrease aspirin-mediated gut permeability. If successful, this project will define a previously undescribed mechanism of cysteinyl leukotriene-mediated allergen absorption in the gut. It also has the potential to elucidate the mechanism underlying the role of aspirin as a cofactor of anaphylaxis. These findings would offer novel therapeutic targets to prevent anaphylaxis in patients with food allergy.

NSF Awards · FY 2025 · 2025-10

Semiconductor technology has been the backbone of modern information technology for more than six decades, driving the growth of numerous emerging technologies such as artificial intelligence (AI), robotics, autonomous driving and 6G. However, compelled by the rapid surge of computing demand, the modern microelectronic devices are facing major challenges in terms of power integrity, thermal stability, and device reliability, leading to significant energy waste on operational margins to sustain the life-time operations of the advanced microelectronic chips. As a result, intelligent runtime chip management with advanced computing methods has becomes a new resolution to address the efficiency and reliability challenges of the next generation semiconductor devices. While AI techniques have recently drawn significant interests for integrated-circuit (IC) design showing high promise, runtime AI assistance for real-time chip management has not been well developed, despite its high potential in overcoming the problems such as power supply noise, chip overheating and device aging. In addition, current AI techniques often observe significant errors when extrapolating beyond its training dataset due to the incompliance with first-principle physics of the semiconductor operations. To fundamentally overcome such challenges, the proposed project will incorporate the emerging physics-informed machine learning techniques with advanced energy-efficient AI accelerators to deliver a new generation of intelligent chip management methods for overcoming the drawbacks of existing solutions such as inaccuracy, long latency and lack of adoptability. The proposed developments will bring fundamental improvements to the modern microelectronic devices in terms of energy efficiency and reliability. This project will collaborate with industry researchers with a goal of delivering the technology for practical industrial applications. By integrating the advanced microelectronic technology with emerging computing methods, the proposed project provides strong educational materials and opportunities for students to learn the multi-disciplinary developments of modern microelectronic design. Course materials and workshops on frontier semiconductor and computing techniques will be developed to provide solid training to the society. This project will develop cross-layer solutions combining both novel AI algorithms and innovative computing circuits to address the ever-increasing challenges in microelectronics including power integrity, thermal management and device reliability. First of all, novel physics informed machine learning models will be developed to support advanced chip management with high accuracy, low latency and high computing efficiency compared with existing solutions; Second, methods of online learning of the developed AI models will be established for intelligent runtime adaptation to deal with the growing impacts of chip variations, workload fluctuations, and operation uncertainties; Third, advanced AI accelerator design with novel computing circuitry will be delivered to provide the most efficient hardware support to the targeted intelligent AI assistive agent for runtime chip management. Real demonstrations with fabricated CMOS test chips will be delivered to showcase the benefits of the proposed techniques in comparison with conventional solutions. Collaborations with industry partners will be performed to evaluate and disseminate the developed technology to practical applications. 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 2025 · 2025-09

Summary Alzheimer’s disease (AD) represents one of the most urgent unmet medical needs of our time. This devastating condition has an enormous impact on the lives of patients, their families and society as a whole. Despite huge research investments, the treatment options remain very few, their efficacy limited, and the occurrence of side effects troubling. This is largely due to the fact that the cellular mechanisms of cognitive impairment in AD and in other dementias remain unclear. The prefrontal cortex (PFC) is critical for higher brain function and goal- oriented behavior. Numerous studies have identified a clear link between cortical electrical oscillations and cognitive tasks, and aberrant oscillations have been observed in multiple cognitive disorders, including AD. Because GABAergic inhibition is critical for oscillations, much attention has been given to the role of excitatory-inhibitory (E/I) feedback in network dynamics, and altered E/I balance is suggested as critical in many neurological disorders characterized by cognitive impairment. Despite the centrality of E/I imbalance in numerous brain disorders, the underlying cellular mechanisms are mostly unknown. E/I imbalance may be due to abnormalities in GABAergic (inhibitory) and/or glutamatergic (excitatory) signaling. Numerous studies suggest that GABA signaling dysfunction underlies reduced brain oscillations and working memory deficits. Accordingly, a recent meta-analysis has identified GABA signaling as particularly vulnerable and a potential therapeutic target in AD. A peculiar biophysical feature of the GABAA current is that the value of its reversal potential is close to the resting membrane potential of most neurons, and thus relatively small variations in the current reversal potential may shift the current effect from hyperpolarizing to depolarizing. The value of the GABAA current reversal potential is mainly regulated by the activity of NKCC1 (SLC12A2) and KCC2 (SLC12A5), two chloride-cation co-transporters that import and export chloride ions, respectively. Numerous recent papers show that depolarizing GABAA current, which is the norm in early development, is present in the adult PFC in diverse brain disorders and may underlie the altered E/I balance in these conditions. Here we hypothesize that depolarizing GABAA current in the PFC of adult animals is a cause of E/I imbalance and cognitive impairment in AD. Accordingly, our preliminary data suggest that restoration of GABAA inhibitory function in the mPFC of an AD mouse model rescues cognitive performance. To test our hypothesis, we will use perforated-patch recordings from acute slices to measure the GABAA current reversal potential, quantitative in- situ hybridization to quantify the expression of NKCC1 and KCC2 RNA in the PFC of APP-Ki NLGF, 5xFAD and control mice, and behavioral assays to determine the effect of pharmacological inhibition of cortical NKCC1 on working and recollection memory in AD mice. Besides the advancement in understanding basic biological mechanisms of cognitive impairment, our experiments may have immediate translational potential, as there are FDA-approved NKCC1 blockers that could rapidly be tested in clinical trials.

NIH Research Projects · FY 2025 · 2025-09

Methane monooxygenase enzymes (MMOs) perform the chemically challenging reaction of the oxidation of methane to methanol. MMOs are metalloenzymes that can either be soluble (sMMO) or membrane-bound (particulate MMO, pMMO). The pMMO is a copper-dependent enzyme that is found in methanotroph intracytoplasmic membranes, organized into hexagonal arrays. pMMO is only active in a lipid environment, indicating the essential role of the membrane in the methane oxidation activity. Previous studies of pMMO in DDM micelles, native lipid nanodiscs, synthetic lipid nanodiscs, and native membranes enabled comparisons of activity in different lipid environments. The goal of this research is to determine why the native membrane is so important for optimal methane oxidation activity using a proteoliposome system. Liposomes provide several advantages over nanodiscs, including increased enzymatic activity as well as the retention of the native array patterning. In Aim 1, pMMO will be reconstituted into liposomes of different sizes and with different lipid compositions, and the effects on activity will be assessed. In Aim 2, the structures of pMMO arrays and their surrounding lipids in liposomes of varying compositions and sizes will be determined via cryogenic electron microscopy. Finally, in Aim 3, the liposomes will be utilized to test the hypothesis that pMMO translocates protons. These combined studies will elucidate how the membrane environment and its components influence the structure and function of pMMO.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Alcohol use disorder (AUD) is one of the most prevalent mental disorders worldwide. Compared to men, women not only suffer from greater alcohol intoxication and withdrawal effects but also develop AUD more rapidly. It suggests that women may be particularly susceptible to the negative valence induced by alcohol withdrawal, which leads to escalated alcohol drinking. However, how valence processing is differentially affected by chronic alcohol drinking between males and females and contributes to the sex differences in the development of AUD is still unknown. A key brain region implicated in valence processing that is dysregulated by alcohol is the bed nucleus of stria terminalis (BNST). Studies show that the BNST exhibits significant sexual dimorphism and is essential for encoding the positive and negative properties of alcohol. In addition, BNST neural activity can be modulated by neuromodulatory actions of neurotensin (NT), a 13 amino acids neuropeptide differentially regulated by sex hormones. Studies have shown that NT enhances inhibitory inputs to BNST neurons and mediates anxiety-like behavior via postsynaptic NT receptor type 1 (NTSR1) and that NT gene expression is correlated with alcohol consumption in women. Together, these findings suggest a sex- dependent NTergic modulation of the BNST on negative affect states during alcohol drinking. We will test the central hypothesis that NTergic modulation mediates the sex difference in alcohol drinking by selectively enhancing BNST encoding of negative valence during acute withdrawal via NTSR1. Aim 1: We will test whether acute alcohol withdrawal enhances BNST encoding of negative valence in females via NTSR1. We will record BNST glutamatergic and GABAergic neurons using in vivo electrophysiology and optogenetic photo tagging in a valence discrimination task before and weekly during the withdrawal periods of intermittent access (IA) alcohol drinking paradigm in both sexes of mice with and without Cre-dependent CRISPR-Cas9 knockdown of Ntsr1 in the BNST. Aim 2: We will examine whether NTSR1 signaling in the BNST drives the escalated alcohol drinking in females by pharmacologically or genetically inhibiting BNST NTSR1 at different stages of the IA paradigm or in glutamatergic and GABAergic neurons of both sexes. Aim 3: We will dissect the NT sources that regulate BNST NT dynamics during valence processing and alcohol drinking. We will measure BNST NT dynamics to motivational valence during a valence discrimination task before and weekly during IA alcohol drinking in both sexes with and without optogenetic inhibition of NTergic inputs to the BNST. We will also examine whether Nts CRISPR knockdown in these projections before IA alcohol drinking affects alcohol drinking of both sexes. Successful completion of this proposal will reveal the functional architecture of NT in mediating dysregulated valence processing in the BNST by IA alcohol drinking and provide critical knowledge of neuropeptidergic modulations in sex differences in AUD.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT There is an urgent need to nominate biomarkers that are likely to predict the efficacy of radiotherapy and accelerate their clinical translation. Efforts thus far have been limited in large part because the omic features regulating tumor cell survival and their frequency across and within individual cancer types had not been studied. Also, radiation is one of the most utilized therapeutic modalities in cancer. Despite this, radiotherapy dose is still delivered using a ‘one size fits most’ paradigm and has not yet lent itself to personalization based on genetic features. Our group of investigators have expertise that spans several aspects of the study of translational lung radiotherapy including advanced functional genomics, use of pre-clinical models of human tumors, personalized therapeutic interventions, bioinformatics, computational biology, and clinical research. Collectively, we have: (1) completed the largest profiling effort of the genetic vulnerability of cancer cells to irradiation to date (533 cell lines); (2) completed an arrayed unary (one mutation per well) gene variant cellular profiling project that has interrogated ~400 common and rare genetic variants for response to ionizing radiation in cells; (3) developed new functional genomic tools to study genome-phenome associations at scale; (4) developed over 500 genetically and clinically annotated patient derived xenograft (PDXs), subset of ex vivo and 3D models for testing using a radiation platform and (5), developed new computational tools and frameworks to personalize radiation dose treatments. The major themes for this proposal are: (i) analyze matched pre- and post-radiotherapy genomes from patient tissue and using PDX to elucidate mechanisms of de novo and/or acquired resistance; (ii) develop predictive diagnostics related to multi-omic tumor variates in patients receiving radiotherapy for lung cancer; (iii) integrate multi-tiered omic data to accurately predict the probability of local failures after lung radiotherapy; and (iv) identify and validate functionally significant molecular lesions in tumors that impact radiotherapeutic efficacy. This project is explicitly designed to utilize patient and model systems data in iterative and complementary testing cycles. Due to our extensive and sustained efforts in this space, we have an established preclinical understanding of the most salient genetic determinants that regulate radiation sensitivity in lung cancer. Altogether, our highly integrated proposal will come together in an unprecedented effort to advance toward clinical integration new information and capabilities that will ultimately improve outcomes for the multitude of patients receiving lung radiotherapy each year in the US and abroad.

NIH Research Projects · FY 2025 · 2025-09

Survival of chronic myeloid leukemia (CML) patients with a sustained major molecular response to Bcr-abl- tyrosine kinase inhibitors (TKIs) may approach age matched controls. Attempts at therapy discontinuation are standard of care for such patients, and 40-50% sustain a therapy free remission (TFR). However, there are no clinical tools that reliably predict TFR duration for an individual. Prior single cell RNA-sequencing (scRNA-Seq) defined a TKI sensitive, proliferative subset of CML-leukemia stem cells (LSCs) and a relatively quiescent subset that persists during TKI treatment. Mechanisms for emergence of these subpopulations are unclear. In preliminary studies, we identified an interaction between CML-LSCs and αβT cells that predicted short TFR in human subjects or a murine CML model. This LSC subset and interacting T cells expanded during post TKI- discontinuation relapse; was not found with sustained TFR; and regressed with a second TKI-induced remission. We identified LSC-interacting T cells as CD8+αβTCR+. To study the impact on TFR, we isolated αβT cell/LSC doublets from mice in TKI-remission, and singlet LSCs from mice with or without doublets by flow cytometry for transplant. Without TKI treatment, recipients of LSC doublets rapidly relapsed, but recipients of single LSCs did not. Adding αβTCR or PDL1 antibody to TKI-treatment decreased doublets and lengthened TFR in recipients. In scRNA-Seq of isolated doublet vs singlet GFP+Lin-ckit+ cells, we found single LSC transcriptomes suggested cell cycle progression and resistance to apoptosis via the unfolded protein response, but doublet LSC transcriptomes suggested HSC quiescence and apoptosis resistance via IAPs. Expression of T cell suppressor genes was increased in doublet LSCs, including PDL1 and Ctla4 ligands, as were cognate proteins on T cells. We hypothesize that interaction of CML-LSC with αβT-cells favors LSC persistence during TKI treatment and provides a source of relapse post discontinuation. We also hypothesize this subset of CML-LSCs suppress αβT-cell immune surveillance, permitting LSC persistence. Conversely, we hypothesize proliferative single LSCs are the source of BCRABL mutations causing TKI-resistance. We will investigate this via 3 Aims; Aim 1: Determine the functional consequences of CML-LSC/αβT cell interaction and identify potential therapeutic targets. We will use a murine CML model to investigate potential mechanisms for interaction defined in preliminary data. Murine results will inform studies of T cell/LSC interactions in human CML cells. Aim 2: Identify mechanisms of LSC-persistence vs TKI-resistance in doublet vs singlet populations. Significance of molecular mechanisms that modulate apoptosis or proliferation in single LSCs vs those in doublets will be studied in mice. Results will be compared to human samples and explored in pre-clinical studies. Aim 3: Define influences of the T cell landscape on TFR and TKI-resistance. ScRNA-Seq data for doublet vs singlet CD8+αβTCR+ cells will be analyzed in humans and murine CML. T cell populations in CML with vs without doublets will be and compared to control subjects to identify influences on doublet formation.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Nausea is a queasy sensation that arises from interoception of physiological distress within the body. However, there remains a critical gap in the fundamental understanding of neurons and neural circuits responsible for this debilitating condition. This proposal focuses on motion sickness (originating in the brain) rather than chemically induced nausea (originating in the viscera), thereby providing an innovative, orthogonal view into the cellular and circuit mechanisms of nausea. The overall objective in this application is to understand how motion-induced nausea is elicited in the brain. The central hypothesis posits that specific neurons in the caudal hindbrain are responsible for the sickness. This hypothesis will be tested in mice by addressing the following three specific aims: 1) Establishing the necessity and sufficiency of discrete subsets of hindbrain neurons for motion-induced nausea; 2) Identifying the genetic profile defining hindbrain neurons associated with nausea; and 3) Mapping the presynaptic to and postsynaptic connections from these neurons. The proposed research is significant because it will reveal the molecules, cells, and circuits responsible for motion-induced nausea. The anticipated outcomes of these studies promise unique and powerful inroads into how the mammalian brain processes distress signals from the body and orchestrates responsive measures.

NIH Research Projects · FY 2025 · 2025-09

Goal: This R34 study will pilot an adaptation for the Mothers and Babies (MB) program for expectant mothers and new parents of infants with Down syndrome (MBDS). Background: Expectant mothers and new parents of infants with Down syndrome are at high risk for perinatal depression. Perinatal depression is both independently, and exponentially associated with long-term adverse neurodevelopmental consequences for infants with Down syndrome. MB is a cognitive-behavioral intervention designed to prevent perinatal depression. MB as one of the two most effective counseling interventions for perinatal depression prevention, with moderate to large effects sizes found across a series of randomized controlled trials (RCTs). However, research suggests that expectant mothers and new parents of children with Down syndrome may have needs that standard MB does not address. Significance: This project will pilot a Down syndrome adaptation to MB, MBDS designed to target mechanisms of grief/loss and social support; and assess whether changes in the target mechanisms are associated with changes in depressive symptoms and parental sensitivity and responsivity to the infant. Innovation: The proposed project is innovative in three ways. First, we plan to conduct the first pilot of a perinatal depression prevention intervention specifically designed for expectant mothers and new parents of infants with Down syndrome. Second, we plan to include fathers to target symptoms of depression, rather than as simply a support person for maternal depressive symptom reduction. Third, we plan to use a group format to establish cohorts of families of infants with Down syndrome of similar developmental stages. Design: Human-centered design and an open trial will inform a subsequent small randomized controlled clinical pilot to test the feasibility of the study protocol in preparation for a larger randomized controlled trial (RCT). Population: Expectant mothers and new parents of infants with Down syndrome. Outcomes: All aspects of the study protocol (e.g., condition allocation, treatment and control condition procedures, data collection, etc.) will be operationalized in preparation for the subsequent RCT. We will assess MBDS effectiveness on target mechanisms of grief/loss and social support; and assess whether changes in the target mechanisms are associated with changes in depressive symptoms and parental sensitivity and responsivity to the infant.

NIH Research Projects · FY 2025 · 2025-09

Housing instability (e.g., difficulties paying rent, eviction) is a significant public policy and public health issue in the U.S., and is associated with 60-70% higher rates of cardiovascular diseases (CVD). One contributor to housing instability experiences such as eviction is a sudden economic shock (e.g., car breakdown) that can have cascading effects resulting in the nonpayment of rent. Recognizing this link, the state of Illinois’ Homeless Prevention Program provides short-term housing assistance (5 months of rental assistance + utilities) to families who are currently housed but who experience an economic shock outside of their control. However, the community organization that administers this subsidy, Connections for the Homeless, has capacity limits that fluctuate across the year (depending on staffing needs and availability). Once capacity has been reached, families who are eligible are turned away. This capacity limit at the community organization level creates a natural randomness in whether eligible families are served or not, allowing us to design a 2-group intervention trial. This natural experiment will test the impact of the housing subsidy among 430 individuals who are housed but who experience an acute economic shock that impacts their ability to pay rent. Effects of either receiving the housing subsidy (n=215) or not (n=215) will be tested on the primary outcome of CVD risk (PREVENT risk equations: risk estimates for total CVD developed by the American Heart Association, based on blood pressure, cholesterol, etc.); the outcomes of housing (moves, evictions, homelessness); and the outcomes of mental health (depression, anxiety), health behaviors (diet; sleep; physical activity; substance use), and biological measures (pro-inflammatory phenotype: immune cells mounting exaggerated cytokine responses to bacterial challenge and becoming insensitive to inhibitory signals from the hormone cortisol; endothelial function, indicated by flow-mediated vasodilation), all of which are important contributors and precursors to CVD. Assessments will occur both at baseline (when households first call to request housing assistance) and 1 year later (after the subsidy has either been received or not). We hypothesize that participants in the housing subsidy group will show lower CVD risk (PREVENT) at 1-year follow-up; will be less likely to have moved, be evicted or become homeless during the ensuing 1 year; and will exhibit better mental health, health behaviors, and biological profiles that are related to CVD risk at 1-year follow-up compared to the group that does not receive the subsidy. Findings from this project could have significant policy implications across multiple domains. Demonstrating that a short-term housing subsidy, provided at a crucial moment in an individual’s life, not only stabilizes housing but also protects cardiovascular health would suggest a high return on investment approach applicable to the rapidly growing population of Americans who experience housing instability.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract The prevalence of Alzheimer's disease and AD-Related Dementias (AD/ADRD) is increasing globally, highlighting an urgent need to identify factors influencing healthy aging and cognitive change. The Health and Retirement Study (HRS) and its sub-study—the Harmonized Cognitive Assessment Protocol (HCAP)—together with their international sister studies, provide invaluable resources for exploring the epidemiology of AD/ADRD. However, there are methodological hurdles and challenges that create barriers for newer researchers to engage with these rich data in a meaningful way. These challenges include issues related to data quality, varying cultural contexts, and harmonizing analyses across datasets. To equip the next generation of researchers with the necessary methodological skills, we proposed to develop M4: Modules on advanced Methodology when Modeling Multinational data. Required modules will focus on the responsible conduct of secondary data analysis, social and behavioral factors related to AD/ADRD, and reproducibility and open science principles. Trainees will choose one or more optional advanced methodology modules appropriate for multinational analyses: item response theory (IRT), coordinated data analyses (CDA), or latent variable alignment methods. Each module will be taught across multiple sessions, allowing for didactic and hands-on training. We have four specific aims: Aim 1: Develop and refine a module-based training program in large-scale data analysis. This is a necessary initial step to create the modules, leveraging and adapting our existing training and tutorials. Aim 2: Instruct trainees in protective behavioral and social factors aimed at reducing AD/ADRD in cross- national contexts. While many different research questions could be addressed with the HRS and HCAP, a primary intention of the proposed training is to address AD/ADRD. Aim 3: Build open science resources to support the identification of factors affecting the trajectory of healthy aging and AD/ADRD. We believe in open science as a primary way to expand knowledge and ensure reproducibility across projects, especially when using widely-available data like HRS and HCAP. Aim 4: Provide ongoing mentorship in advance methodologies. By engaging trainees across modules, we can develop mentoring relationships whereby we can provide ongoing support in their mastery of these advanced methodologies. M4 will be offered primarily by synchronous virtual learning. Then it will be adapted for asynchronous self-paced learning, allowing participants to engage with it at their convenience. This is of vital importance, insofar as these are multinational data, and international researchers need to be equipped to analyze their own data. Finally, abbreviated versions of the modules will be offered as in-person pre-conference workshops to raise interest and enhance trainee recruitment, ultimately aiming to foster a robust community of researchers dedicated to advancing the understanding and reduction of AD/ADRD.