University Of Wisconsin-Madison

NIH Research Projects · FY 2026 · 2023-06

PROJECT SUMMARY/ABSTRACT Advances in early detection of Alzheimer’s disease (AD) pathology via imaging and fluid-based biomarkers in individuals with little to no evidence of cognitive decline allow for the targeted enrollment of participants who are most likely to show benefit from early interventions. However, many of these biomarkers are costly and invasive. As a result, more accessible, performance-based measures of cognitive function are needed to improve early detection, aid in recruitment for clinical trials, serve as clinical endpoints for interventions, and improve access to individuals from underserved communities. In this R01 proposal, we respond to this need: by leveraging existing longitudinal data of over 3,000 speech samples from two large cohorts, we propose validation of a digital speech marker across all ADRD stages. Connected speech and language (CSL) analysis of digitally recorded speech—the detailed measurement of everyday spoken language—is a noninvasive digital marker that measures communication, an activity essential for quality of life. Advances in natural language processing and Machine Learning have made automated linguistic analysis a rapid and effective means for analyzing CSL, while the ubiquity of mobile devices makes remote and frequent digital speech collection widely accessible to more people at risk for AD. Our central objective is to further develop, increase accessibility, validate, and improve the sensitivity of digital speech markers to AD through new analytic approaches and remote collection methods. By pursuing the following specific aims on a large-scale dataset, we investigate early changes in CSL to predict how and when CSL relates to the risk of developing ADRD: Aim 1: Validate connected speech and language measures from older adults across multiple stages of ADRD, including prodromal ADRD, mild cognitive impairment, and dementia. Aim 2: Test the hypothesis that connected speech and language measures are associated with AD biomarkers, including Aβ and tau from PET imaging, levels of tau, Aβ, and neurodegeneration (NFL) in CSF and blood plasma, and global neurodegeneration from MRI. Aim 3: Evaluate the usability and accessibility of frequent, remote at-home speech collection and validate linguistic metrics collected remotely against those obtained in person. The results of these three aims will lead to a larger goal of understanding the CSL features that are sensitive to cognitive decline and AD neuropathology in at- risk adults, and the linguistic and acoustic markers necessary to develop a widely accessible tool measuring early cognitive change along the ADRD continuum.

NIH Research Projects · FY 2026 · 2023-06

Project Summary A major cause of male infertility is defective and atypical development of the Wolffian duct, the embryonic structure that give rise to male internal reproductive tract organs. It is known that Wolffian duct differentiation is predominantly driven by the action of the testis-derived androgens. However, how the androgen signaling coordinates the process of the Wolffian duct differentiation remains unclear in the field of reproduction. The androgen action in Wolffian duct differentiation is mediated by the androgen receptor (AR), which is expressed in both Wolffian duct epithelium and mesenchyme. Using a new mesenchyme-specific Ar knockout mouse model, we provided the first genetic evidence that the AR in the mesenchyme is essential for Wolffian duct differentiation. By comparing chromatin accessibilities and transcriptomes of Wolffian duct mesenchymes from female and male embryos, we discovered a set of androgen- induced chromatin accessible regions and a new androgen-induced mesenchymal factor R- Spondin 3 (Rspo3). RSPO3 is a WNT signaling activator secreted from the mesenchyme to activate epithelial Wnt signaling that is essential for Wolffian duct morphogenesis. While the mesenchyme governs epithelial differentiation, the epithelium has the reciprocal inductive effects on the mesenchyme by synthesizing a paracrine growth factor WNT9B. We found that the loss of Wnt9b caused Wolffian duct degeneration at the sexual differentiation window when the androgen signaling was supposed to promote Wolffian duct survival. These observations lead to and support our central hypothesis: the androgen-dependent Wolffian duct differentiation requires the stimulation of the epithelium-derived WNT9B, and the androgen signaling in Wolffian duct differentiation is mediated by the mesenchymal AR and executed by the androgen-induced mesenchymal factor RSPO3 via epithelial-mesenchymal interactions. We will determine the mechanisms of WNT9B, AR, RSPO3 actions in promoting Wolffian duct differentiation by utilizing a combination of tissue-specific gene knockouts, gene expression assays, fluorescence-activated cell sorting, RNA-seq, ATAC-seq, and single cell mRNA-seq. AR and WNT9B variants in humans have been associated with defective androgen-dependent male reproductive tract differentiation. Therefore, the completion of our proposal will not only yield fundamental knowledge of basic mechanisms underlying androgen-dependent Wolffian duct differentiation but also provide knowledge directly relevant to our understanding of disorders of male sexual development in humans.

NIH Research Projects · FY 2026 · 2023-06

Abstract Myelination of axons in the nervous system is critical for not only conduction of action potentials, but also for providing tropic support to ensure long term survival of neurons in both the central and peripheral nervous systems. Myelin disorders are a major cause of neurological disease, and can be caused by genetic disorders, infectious disease, and inflammation. The peripheral nervous system has substantial plasticity in being able to regenerate after nerve injury, and critical transcription factors and their target gene networks have begun to be elucidated. Schwann cell reprogramming to a new differentiated state is a critical and rate limiting factor in peripheral nerve regeneration, particularly when regeneration is impaired as a function of aging/chronic denervation. Therefore, understanding the pathways that control gene expression reprogramming will provide insight into means by which remyelination after nerve injury can be accelerated. The long term objective of our laboratory is to elucidate an integrated mechanism of Schwann cell reprogramming after nerve injury based on critical transcriptional and epigenomic switches. We have found that many critical injury genes are associated with polycomb-associated histone modifications (H3K27me3 and H2AK119ub) prior to injury, and this proposal focuses on testing how reversal of the polycomb pathway is required for Schwann cell responses to peripheral nerve injury. We have also found an expected role of polycomb eraser proteins in myelin homeostasis. Using a variety of techniques established in our laboratory, we will test for the first time the involvement of modulating PRC1 pathway in nerve injury responses. In addition, our work has highlighted mechanisms by which Sonic Hedgehog is activated in repair Schwann cells, and we will employ novel mouse resources to elucidate the mechanisms and role of Sonic Hedgehog signaling in injured peripheral nerve.

NIH Research Projects · FY 2025 · 2023-06

PROJECT SUMMARY/ABSTRACT Individuals with Alzheimer’s disease and related dementias (ADRD) experience progressive cognitive decline and behavioral and functional impairment. With disease progression, individuals with ADRD also experience heightened risk for poor outcomes which often culminate in the need for skilled care provided in nursing homes (NH); almost all individuals with ADRD will receive short- or long-term NH care at some point. While providing high-quality, person-centered ADRD care is a national priority, progress toward this goal is limited in part by a lack of understanding regarding the influence of major comorbidities on ADRD disease burden, care needs, and symptoms. In particular, comorbid serious mental illness (SMI), which is common among individuals with ADRD, remains understudied despite its impact on disease risk, burden, care needs, and symptomatology. Burden of both ADRD and SMI is particularly high in NH settings. Understanding whether SMI plays an additive role in shaping behavioral symptom presentation among individuals with ADRD, care needs (e.g. pain), and outcomes (e.g. falls) can inform intervention targets and improve person-centered care across settings. Towards this end, I propose a mixed methods sequential explanatory design to investigate the contribution of SMI to ADRD symptoms, outcomes, and care interventions; domains informed by the Need-Driven, Dementia-Compromised Behavior Model. I will utilize secondary data available through the Minimum Data Set 3.0 (MDS 3.0) (Aim 1/1A) and conduct interviews with NH staff (Aim 2). My long-term goal is to establish a program of research designing and delivering person-centered interventions for symptom management among adults with ADRD and complex comorbidities. My short-term objective is to characterize ADRD/SMI’s relationship to shared and distinct symptomology, outcomes, and care interventions; and implications for individuals with ADRD and provision of person-centered care. Under Aim 1, I will describe patterns and characteristics of comorbid ADRD/SMI diagnosis among NH residents using 2019 MDS 3.0 NH assessment data (N~500,000). Applying clustering techniques, I will define underlying ADRD/SMI disease co-occurrences and their variation by demographic characteristics. I will then evaluate how distinct ADRD/SMI comorbidities are associated with unique behavioral symptoms, outcomes (e.g. falls, functional status) and care needs/interventions (e.g. medication use, restraint use) as compared to ADRD alone (Aim 1A). Under Aim 2, I will explain and extend Aim1/1a findings by conducting content analysis of interviews with NH staff (e.g. nurses, nursing assistants, social workers, providers) (N=30, total) about their perceptions of ADRD/SMI comorbidities and care of residents with complex cognitive and psychiatric care needs. Impact: Findings from the proposed study are responsive to ADRD research priorities and will provide foundational data informing targets for person-centered care interventions for patients with ADRD/SMI by illuminating the contributory role of SMI to ADRD symptoms and care. The training plan will support my broader career goal of becoming a leading clinician scientist focused on aging and ADRD care.

NIH Research Projects · FY 2026 · 2023-06

Project Summary: The operation of RNA polymerases (RNAPs) relies on numerous conformational changes. During eukaryotic transcription, RNA Polymerase II (Pol II) encountering oxidative lesions in its DNA template often leads to misincorporation and transcriptional stalling. These events contribute to tumor growth in skin cancer. Mycobacterium tuberculosis (Mtb) causes lethal tuberculosis and is responsible for over 1 million deaths per year. Transcription initiation complexes of Mtb RNAP, especially the DNA loading gate, are effective targets for the development of antibiotics. Revealing the dynamics of transcription initiation can thus provide novel mechanistic insights into prokaryotic transcription and greatly facilitate the understanding of inhibition mechanisms for antibiotics targeting Mtb RNAP. These two important biological problems in transcription drive us to develop novel methodology using the generalized master equation (GME) to model biomolecular conformational changes. My group has been successful in developing GME methods that explicitly consider the memory functions of biomolecular dynamics and outperform the popular Markov State Model (MSM) method. However, as an emerging approach, the current implementation of GME is prone to instability when estimating memory functions for complex RNAP systems. We here propose novel methods to build GME models. Our specific aims are: 1. To develop new GME methods to model conformational changes. Specifically, to derive a new theory (IGME) to solve the GME, to develop efficient implementations of the GME to enhance numerical stability when computing memory kernels from molecular dynamics (MD) simulation trajectories, and to create a protocol tailor-made for building GME models to study biomolecular conformational changes. Our preliminary work shows that the proposed IGME method greatly outperforms the original implementation of GME in yielding robust and accurate predictions of the biomolecular dynamics, especially for the complex RNAP system. 2. To reveal how the dynamic coupling of several key conformational changes (i.e., the loading of NTP, the rotation of the damaged DNA base, and the translocation of Pol II on the DNA template) leads to transcriptional mutagenesis and/or stalling. Specifically, to construct GME models to elucidate molecular mechanisms of 8-oxo- guanine (8OG) and Guanidinohydantoin (Gh) lesions induced ATP misincorporation and/or transcriptional stalling. 3. To elucidate the molecular mechanisms of transcriptional initiation and its inhibition of Mtb RNAP. Specifically, to construct GME models to reveal the dynamics of the Mtb RNAP’s loading gate without DNA, and to further reveal the dynamics for the transition from a partially formed transcription bubble to a fully formed bubble, a conformational change involving both Mtb RNAP’s gate opening and DNA unwinding. We further aim to understand the recognition mechanisms of multiple antibiotic compounds, including Myxopyronin (Myx) and Fidaxomicin (Fdx) that target the loading gate motion, and Sorangicin (Sor) that inhibits the formation of the full transcription bubble. These mechanistic insights will facilitate the rational design of new inhibitors fighting drug resistance of Mtb in the long term. Throughout our studies, we will work closely with our experimental collaborators to conduct biochemical, time-resolved X-ray, and Cryo-EM experiments to test and validate our predictions. Our innovative GME methods will provide a general computational framework to model functional conformational changes of biomolecules. Our developed protocol and associated code development in the MSMBuilder software will widely benefit the biophysics community.

NIH Research Projects · FY 2026 · 2023-06

ABSTRACT Current intensive care unit (ICU) care delivery for acute respiratory failure fails to meet the needs of patients near the end of life, their families, and their interprofessional ICU teams. Efforts to improve end-of-life ICU care have focused on communication between families and clinicians. Yet, poor end-of-life outcomes for patients, families, and ICU teams remain intractable, in part because of the system-level processes and norms that drive ICU care delivery. Time-limited trials have promise to disrupt system-level drivers of poor quality end-of-life care and establish processes of care delivery that better support patients, families, and ICU teams. A time- limited “trial” is an agreement among patients, their surrogate decision makers, and clinicians to attempt life- sustaining treatment for a predefined period and then re-evaluate its utility. This patient care model was proposed by palliative care experts to address prognostic uncertainty, and navigate competing desires simultaneously held by many patients—to extend life when possible and to avoid prolonged life-sustaining treatment if the chance of recovery is low. Time-limited trials have been described in the literature, endorsed by palliative and critical care professional societies, and are currently used in ICU practice. Yet, time-limited trials remain poorly formalized in practice with little evidence to guide ICU clinicians in how to conduct trials. The central objective of this study is to define the optimal care delivery processes of a time-limited trial in acute respiratory failure. This application responds to NHLBI’s Notice of Special Interest in Palliative Care (NOT-HL- 20-737), including the focus on models of ICU-based palliative care for acute respiratory failure and on how ICU structure and culture influence palliative care delivery. This project is a sequential mixed methods, prospective observational study in five ICUs where time-limited trials are currently used. In Aim 1, focused ethnography of 50 time-limited trials in patients with acute respiratory failure will be used to map current processes of time-limited trial care delivery for patients with acute respiratory failure. In Aim 2, a prospective, observational cohort study in 5,810 patients with acute respiratory failure will be used to elucidate the relationships between time-limited trial processes and end-of-life outcomes for patients, surrogates, and ICU teams. Aims 2a and 2b will test whether time-limited trial processes are associated with better end-of-life outcomes and Aim 2c will characterize mechanisms by which processes influence end-of-life outcomes. This work will transform time-limited trials from a promising model into a clearly defined and optimized care delivery process. This process will be immediately actionable to guide frontline ICU teams currently using or considering time-limited trials. This study will also establish the necessary foundation for the design and testing of complex health interventions that support time-limited trials and safeguard against unintended harms.

NIH Research Projects · FY 2025 · 2023-06

PROJECT SUMMARY/ABSTRACT Frontline treatments for major depressive disorder (MDD), including cognitive behavioral therapy (CBT), have limited effectiveness, with ~½ of patients not responding to CBT and ½ relapsing within 2 years. Strategies that could increase the potency of MDD treatments are needed. We have previously shown large psychological and neuroplasticity improvements for 75 minutes post-exercise in adults with MDD. This creates a neurologically fertile post-exercise window with the potential to maximize the effects of CBT performed immediately after exercise. We have developed a novel treatment strategy using aerobic exercise to prime a subsequent CBT session (‘ActiveCBT’). Exercise priming is hypothesized to lead to greater engagement of CBT mechanisms of action through both common (e.g., working alliance) and specific (e.g., behavioral activation) factors linked to depression outcomes; this CBT augmentation approach has yet to be empirically tested. Our published and preliminary work show exercise acutely improves psychological (eg, state anhedonia) and neuroplastic (eg, BDNF) markers in MDD, and our n=10 pilot feasibility trial of this project indicates high likelihood for logistical success and plausible effects of exercise priming on target mechanisms. The R61 will consist of an 8-week randomized controlled trial in 40 adults with MDD performing 30-minutes of moderate exercise or quiet rest (ActiveCBT vs. CalmCBT) prior to weekly CBT sessions. Using a time- and attention-matched control, all participants will view a standardized documentary series for these 30 pre-therapy minutes with the only difference between groups being exercise. This project will also employ cutting-edge, machine learning and natural language processing via the Lyssn platform to objectively evaluate therapy. The overall R61 goals are to: 1) demonstrate exercise priming effects on target CBT mechanisms (working alliance and behavioral activation), 2) link psychological and neuroplastic exercise priming to CBT mechanisms, and 3) explore sensitivity of objective machine learning-based markers from Lyssn to ActiveCBT-induced therapy differences. Upon identification of a moderate between-group difference in working alliance and/or behavioral activation favoring ActiveCBT, the R33 will proceed with a 2:1 randomized, controlled trial (n=100), comparing ActiveCBT to CalmCBT. The overall R33 goals are to: 1) confirm effects of exercise priming on target CBT mechanisms, and 2) determine the preliminary efficacy signal of exercise priming, and determine the strength of the efficacy link with CBT mechanisms. Our working hypotheses are that we will confirm higher working alliance and/or behavioral activation in ActiveCBT, with greater depression effects at 8 weeks and 12 weeks later linked to greater working alliance and/or behavioral activation. These trials will establish the potential for a subsequent confirmatory efficacy trial to rigorously test the potential of exercise priming to enhance treatment. If this innovative, biologically-informed approach successfully ‘primes’ subsequent therapy, this could be used to augment other treatments and in other disorders, significantly improving mental health treatment.

NIH Research Projects · FY 2025 · 2023-05

PROJECT SUMMARY/ABSTRACT – OVERALL The genome editing community is celebrating the ten-year anniversary of landmark CRISPR papers in 2022. What is notable to many in the field is tremendous technological advances in editing precision and versatility, along with a Nobel Prize and billions of dollars of investment. However, there is a palpable sense that these advances have not been translated into valuable drugs at a sufficient rate. In vivo gene editing still faces substantial challenges, especially when it comes to safety, efficacy, and delivery. While the development of EDIT-101, a viral vector carrying Cas9 to treat a rare retinal disorder (BRILLIANCE trial), provided a clear path to the clinic for a genome editing therapeutic program, this path can be challenging to follow for others in the field interested in developing human therapeutics. Viral strategies have several limitations involving an immune response to vector elements and prolonged expression of the editor for the lifetime of the patient, heightening off-target concerns. Newer editors like base editors cannot be readily packaged into common viral vectors, plus there is a challenging supply chain for viral vector manufacturing. We seek to overcome the limitations with viral delivery systems by using novel nonviral delivery systems termed the Silica NanoCapsule (SNC) and Target Active Gene Editors (TAGE). They can efficiently deliver genome editors to the retina with high efficiency, reaching levels of 10-70% that are among the best for nonviral delivery of editors to the eye and comparable to viral delivery systems. Based on our strong published and preliminary data we propose to develop nonviral gene editing products to treat Best Disease (BD) and Leber Congenital Amaurosis (LCA), two diseases affecting ion channels of the retinal pigment epithelium (i.e., RPE channelopathies). Our team, spanning academia and industry, will pursue the following aims. In Overall Aim 1, we evaluate the clinical readiness of a genome editor targeting a post-mitotic cell through transient, localized, and nonviral delivery. To date, outside of the liver, only viral editors have reached an IND for in vivo editing. Here, we rigorously evaluate the potential of our SNC and TAGE nonviral delivery systems for the treatment of LCA and BD. We invest our most significant effort into the Lead Project 1 that develops a base editor within a SNC. This project will reach an IND within five years and provide synergy for other projects. In Overall Aim 2, we create a platform that can address many rare diseases of the eye by streamlined manufacturing of different guides with a simple pipeline for preclinical testing. In Overall Aim 3, we provide the SCGE Consortium and broader genome editing field a set of regulatory interactions that clarifies development path to the clinic for new gene editing therapies. Finally, through our experience with BD, we expect to learn about the regulatory path in developing somatic cell genome editors for scenarios when no suitable animal model exists. Because the majority of known pathological mutations in the eye have no suitable animal models, sharing this knowledge will have a large impact on subsequent genome editing leads.

NIH Research Projects · FY 2026 · 2023-05

An estimated 1/3 of the world population is already infected with Mycobacterium tuberculosis (M. tb), the causative agent of tuberculosis (TB), with an almost 5-10% of latent infected patients developing active tuberculosis during their lifetime. In the last three years, infection with severe acute respiratory syndrome-Coronavirus-2 (SARS-CoV-2, a.k.a. SCV2) the causative agent of COVID-19, has resulted in 6.5 million deaths. According to the WHO Global TB Report 2022, the SCV2 pandemic in 2020-2021 was associated with the reversal of the steady decline in TB death rate for the first time since 2005. Our long-term goal is to better control tuberculosis and manage cases of superinfection with SCV2 as well as gain insight to immunological consequences of SCV2 infection. Preliminary data from our group indicated SCV2 superinfection increased M. tb loads in murine lungs and promoted dissemination to extra-pulmonary organs, which was associated with specific changes in immunomodulatory cytokines (e.g., IFN-g and IL10) and decreased general inflammation in the lungs. These data underpinned our central hypothesis that SCV2 superinfection compromises anti-TB immunity, leading to greater dissemination of M. tb. To address the central hypothesis we plan to I) Characterize the TB/SCV2 superinfection in a latent murine model of TB using C3HeB/FeJ mice in a setup termed Immune Competent TB Model (ICTM); II) Define mechanisms of M. tb dissemination during SCV2 superinfection by targeting IFN-g and IL-10 cytokines using reporter and repletion murine models that are infected with M. tb before superinfection with SCV2 BA.5. Finally, III) Examine M. tb progression in Diversity Outbred (DO) mouse model compared to inbred mice to characterize mycobacterial growth phase associated with pathological and immunological changes during TB/SCV2 superinfection. Outcomes from this project will further improve our understanding of TB immunopathogenesis and how latent infections in humans can convert to active cases of TB including furthering knowledge on the granuloma’s role in containment and pathogenesis. Importantly, gained knowledge could help in understanding TB immunopathogenesis during any future respiratory superinfection, not SCV2 alone. Future research building on outcomes from this project could help in the control of both TB and COVID-19 pandemics that could be tailored for other respiratory emerging infections (e.g. Influenza).

NIH Research Projects · FY 2026 · 2023-05

PROJECT SUMMARY Dysphagia frequently develops in persons with AD/ADRD and leads to serious health consequences, including increased caregiver burden, malnutrition, pneumonia, decreased quality of life, and mortality. Targeted, efficacious rehabilitative interventions for dysphagia have been developed in other neurodegenerative populations, but there are currently no effective treatments for dysphagia in AD/ADRD that have lasting effects on swallowing physiology. Prior to developing interventions to prevent dysphagia-related adverse sequelae in patients with AD/ADRD, in-depth understanding of factors contributing to dysphagia risk and trajectories of swallowing change across disease progression is urgently needed, especially in early stages when interventions can be most effective and impactful. Small, cross-sectional studies in AD/ADRD have suggested changes in swallowing beginning early in disease progression. However, these prior studies examining dysphagia in AD/ADRD lack the comprehensive and longitudinal characterization of swallowing function across the dementia continuum necessary to improve clinical management. Additionally, while novel, multi-modality swallowing assessments have enabled detection of subclinical swallowing impairments in other neurogenerative populations, the nature of subclinical changes in AD/ADRD remains unknown. To address these critical knowledge gaps, we propose a prospective cohort study of persons with AD/ADRD stratified by disease stage (very mild to moderate) and their care partners. Participants will undergo comprehensive dementia characterization, including neuropsychological testing and plasma-based biomarkers, as well as multi-modal swallowing assessments at baseline and every six months. We will also collect a variety of clinical factors (e.g., demographics, comorbidities) and measures of oral function (e.g., saliva production, oral health, lingual pressures), respiratory pressures, sarcopenia, and nutritional status at baseline. The overarching goal of this research is to inform development of intervention strategies for persons with AD/ADRD through enhanced clinical and epidemiologic understandings of the onset, nature, and progression of swallowing deficits. Our multidisciplinary team of experts will achieve this objective via the following specific aims: 1. Define swallowing changes from very mild to moderate AD/ADRD to: 1a) characterize subclinical dysphagia features; and 1b) identify clinical factors and swallowing measures that contribute to symptom reporting; 2. Identify risk factors for swallowing dysfunction from very mild to moderate AD/ADRD; and 3. Determine trajectories of change in swallowing function in persons with AD/ADRD and association with risk factors. The proposed work represents the first longitudinal, comprehensive study of swallowing function in a well-characterized cohort of persons with AD/ADRD. Improved understanding of swallowing function across disease progression will inform development of targeted interventions to address identification, prevention, and rehabilitation of dysphagia in AD/ADRD.

NIH Research Projects · FY 2026 · 2023-05

Project Summary Currently, there is no effective way to slow down the progress of Alzheimer’s disease (AD) or prevent it. CRISPR genome editing is a revolutionary and versatile genetic engineering technique, making it possible to treat the root causes of genetic neurodegenerative diseases (NDDs) such as AD. However, the promise of brain gene therapy relies on the efficient delivery of biologics to the brain, which is extremely challenging due to the blood- brain barrier (BBB). To date, in vivo brain gene therapy has mostly been achieved using viral vectors that require laborious customization and have troublesome safety profiles. Non-viral vectors are largely administered via intracranial administration, which is invasive and can only enable gene therapy in a small and localized brain region. Similar to other NDDs, AD affects multiple brain regions. Thus, there is an urgent need to develop efficient non-viral delivery vehicles capable of bypassing the BBB for safe and efficient brain-wide gene therapy. The objectives of this project are (1) to engineer glutathione (GSH)-responsive silica nanocapsules (SNCs) capable of bypassing the BBB and delivering CRISPR genome editors to the whole brain systemically, and (2) to evaluate the therapeutic efficacy and biosafety of brain-wide genome editing enabled by the optimized SNC for the treatment of AD using a novel amyloid precursor protein (APP) knock-in AD mouse model and a unique gene target for APP modulation. The unique SNC possess a long list of desirable properties including versatile payload types, versatile surface chemistry for ligand conjugation, high payload loading content and efficiency, small particle sizes, excellent in vivo stability, good biocompatibility, and scalable production. Our preliminary data has shown that intravenously administered SNCs can efficiently deliver mRNA, DNA, Cas9 mRNA/sgRNA, and Cas9/gRNA ribonucleoprotein (RNP) to the whole brain of healthy mice with intact BBB. We aim to further optimize the amounts of the dual brain-targeting ligands (i.e., glucose and rabies virus glycoprotein (RVG) peptide) and dosages of the SNCs for enhanced brain-wide systemic delivery of two types of CRISPR genome editors (i.e., (1) Cas9 mRNA/sgRNA, and (2) plasmid DNA with a neuron-specific human synapsin 1 (SYN1) promoter and expressing both Cas9 and sgRNA). We will further determine their therapeutic efficacy and biosafety in treating AD using a novel APP knock-in AD mouse model while employing a unique gene target for APP modulation. The gene editing efficiency and biosafety profiles of the SNC, and the pathological and behavior changes of the AD mice will be monitored by various techniques including a novel serial two-photon tomography whole-brain amyloid imaging platform. With promising initial studies, the best- performing SNC will be submitted to the Preclinical Testing Core of the NIA-sponsored STOP-AD program for comprehensive preclinical evaluation. This project will pave the road for a new, safe, non-invasive and effective therapeutic approach for familial AD. Given the modularity and versatility of the SNCs, and ease of targeting different genes by the CRISPR system, we anticipate that our SNCs will be applicable for a wide range of NDDs.

NIH Research Projects · FY 2026 · 2023-05

PROJECT SUMMARY Poly-(ADP ribose) polymerase (PARP) inhibitors as a drug class are considered one of the great success stories of Precision Medicine with Food and Drug Administration drug approvals accumulating for growing proportions of patients with diverse cancer types. As well, preclinical studies are investigating their use in a multitude of non-malignant diseases. These drugs inhibit multiple PARP enzymes at once, some of which are critical enzymes for sensing and coordinating repair of several types of DNA damage. Further, these drugs trap the PARP1 enzyme onto DNA, resulting in a DNA double strand break to resolve this lesion. Thus, by design, these drugs increase DNA damage in cells, raising concern that they could be toxic to or even mutate normal cells that rely on PARP enzymes for their functions. Thus, clinical trial and drug safety databases showing serious bone marrow adverse effects, including severe, and sometimes irreversible lowering of the blood counts, and an increased risk of deadly therapy-related bone marrow cancers suggest that hematopoietic stem and progenitor cells are uniquely susceptible to PARP inhibitor effects. However, mechanism and causation are not yet clear. The long-term goal is to understand normal bone marrow function and if patients with germline or acquired mutations in their bone marrow cells are uniquely susceptible to the adverse effects of specific exposures. The central hypothesis is that 1) PARP inhibitors, due to their ability to stall replication forks and cause double strand DNA breaks, impair hematopoietic stem and progenitor cell proliferation, resulting in decreased blood cell production from the most proliferative subsets more than others., and that 2) these PARP inhibitor effects mobilize long-term stem cells to help meet blood production demands and at the same time create a competitive environment in which stem cells with TP53 or other mutations are favored to outgrow wild- type, but ultimately predisposing patients to bone marrow failure or leukemia. The rationale for this project is that an ever-expanding group of patients are being treated with PARP inhibitors and many experience serious blood and bone marrow toxicities, including development of therapy-related blood cancers, but the mechanism and methods to prevent these serious toxicities are not known. Using mouse and human in vitro and in vivo models, we will test these hypotheses with the following specific aims: 1) Determine how PARP inhibitors disrupt normal hematopoiesis to cause cytopenias; and 2) Determine whether PARP inhibitors create a selective environment and unmask fitness differences in hematopoietic stem and progenitor cells with varying genotypes or are able to mutate these cells on their own. The research proposed in this application is innovative because it is the first study to determine how inhibiting multiple PARP enzymes simultaneously impacts normal hematopoiesis on a mechanistic level. This proposal is significant because it will allow identification of prevention strategies for safe PARP inhibitor use and of new drug targets for improving hematopoiesis via PARP enzyme manipulations.

NIH Research Projects · FY 2026 · 2023-05

Understanding the role of malleable epigenetic mechanisms in birth defects is a direct path to prevention strategies. Orofacial clefts (OFCs) of the lip and palate are among the most common human structural birth defects, affecting 1 in 800 newborns, and pose serious individual, familial, and societal burdens. Prevention strategies for OFCs are elusive because our current understanding of causative factors is inadequate. Epidemiologic and traditional genetic studies have shown that OFCs are etiologically complex outcomes that result from multifactorial genetic and environmental influences. Epigenetic mechanisms are an exciting new focus in understanding the genesis of OFCs because they mediate the effect of environmental influences on the genome during sensitive embryonic periods. Our proposal specifically focuses on DNA methylation because this epigenetic mechanism is environmentally sensitive and a practical target of prevention and therapeutic strategies. While implicated by multiple lines of evidence, the biological role of DNA methylation in orofacial development is unclear. We have established novel models and generated key proofs of concepts that poise us to uncover how DNA methylation regulates orofacial morphogenesis and to define the role that DNA methylation plays in modulating OFC susceptibility. In this project, integrated genome-wide methylation and bulk and single-cell transcriptome approaches will be applied to define molecular and cellular mechanisms of OFC pathogenesis resulting from disrupted DNA methylation in the cranial neural crest. The role of DNA methylation in multifactorial OFC susceptibility will then be defined by integrating multiple environmental and dietary modulators of DNA methylation to genetic (Wnt9b KO) and chemical (Shh antagonist) mouse models of incompletely penetrant OFCs. Finally, epigenome editing will be applied to evaluate the functional impact of OFC-associated methylation changes on gene expression and cranial neural crest biology. Completion of the proposed studies will bring fundamental insight into how DNA methylation regulates cranial neural crest biology and orofacial morphogenesis. By defining environmental- and dietary-mediated methylome- transcriptome responses that alter OFC susceptibility, these studies will also provide a necessary foundation for identification of environmental influences that modulate DNA methylation and contribute to OFC risk. Pursuing this line of investigation will advance our long-term goal of developing prevention strategies for etiologically complex birth defects by identifying culpable environmental influences and defining their mechanisms of action.

NIH Research Projects · FY 2026 · 2023-05

Cerebral ischemia significantly alters the expression and/or function of transcriptional and translational mechanisms including various classes of noncoding RNAs, epigenetics and epitranscriptomics. My research in the past 20 years evaluated these mechanisms that are central in promoting either secondary brain damage or recovery after stroke with a goal to design novel therapies. In the 7 years, I will focus on studying the role of various RNAs in promoting post-stroke brain damage. The goal is to understand the mechanisms as well as identifying new therapeutic targets to minimize the secondary brain damage and to promote functional recovery after stroke. In this R35, I propose 4 projects. Project 1: In a currently funded RO1 grant we are evaluating the functional significance of an epigenetic modification called DNA hydroxymethylation (5hmC). Our studies so far showed that stroke leads to induction of 5hmC in many prosurvival genes that induces their expression. We intriguingly observed that many lncRNA induced after stroke also show increased 5hmC levels. In this project, we will continue the studies to understand the significance of 5hmC induction in lncRNAs to post-stroke functional outcomes. Project 2: RNAs can be tagged by >150 distinct chemical modifications, which are collectively defined as epitranscriptomic modifications that form an additional layer of post- transcriptional gene regulation. Among them, methylation of the adenosine at N6-position (N6- methyladenosine; m6A) is the most abundant modification in the brain. Our studies show that focal ischemia downregulates m6A demethylase FTO leading to increased abundance of m6A-tagged mRNAs. Many of these are inflammatory and apoptotic. Furthermore, activation of FTO promotes RNA demethylation and neuroprotection. We will use molecular tools to evaluate the functional significance and mechanisms of increased m6A methylation of RNAs in brain damage after stroke. Project 3: RNAs can also undergo the epitranscriptomics modification of glycation and the neural glycoRNAs interact with microglial Siglec receptors to dampen inflammation following CNS insults. We will study the significance of glycoRNAs and the mechanism of action in brain after stroke. Project 4: Various classes of RNAs collaborate in their actions. We particularly observed that circular RNAs interact with lncRNAs to modulate mRNAs and microRNAs. We will analyze the role of RNA networks in relation to ischemic brain damage using 2 examples. Project 4A: The first one is the interaction of a circRNA (circPUM1) with a lncRNA (NORAD) to control a mRNA that codes an RNA binding protein (Pum1) and a mRNA that codes BNIP2. Project 4B: An ongoing RO1 is studying the role of miR-7 in controlling -Synuclein in post-stroke brain. Although mature miR-7 levels decrease after stroke, levels of premiR-7a and 7b are not altered. As circRNA CDR1as binds and stabilizes miR-7, we are planning to study the interactive role of this CDR1as-miR-7--Synuclein in mediating post-ischemic brain damage. Overall, the above projects are all focused to evaluate the significance of various RNAs with a goal to find new mechanisms and new targets to design future molecular therapies to curtail post-stroke brain damage.

NIH Research Projects · FY 2026 · 2023-05

Summary/Abstract The 2016 report issued by the Surgeon General’s Office, “Facing Addiction in America,” the first of its kind issued by the Surgeon General, brought into stark relief the major impact of substance misuse and use disorders and the need to rethink how prevention and treatment services are currently delivered in the United States. While isolated aspects of substance misuse are improving, such as tobacco cessation and opioid prescribing practices, enormous service gaps persist that allow the ongoing and enormous negative impacts of risky and problem substance use. Further, these negative impacts put a particular burden on gender and ethnic minorities and other disadvantaged populations and communities. The isolation of specialist substance abuse treatment from mainstream medicine, the report further points out, presents a barrier to a potentially more effective public health model to reduce substance-related harms. If this landscape of epidemic substance misuse, use disorders, and related conditions is to improve and patient and public health and safety outcomes enhanced, prevention and treatment services delivery require rethinking. The Addiction Health Services Research conference, held since 2005, has annually brought together researchers, policy makers, and treatment providers to focus upon how such systems redesign might most effectively be implemented and sustained. In the current application, we propose to enhance this conference by specifically focusing on improving the well-being of gender and ethnic minorities and other marginalized groups struggling with risky and problem substance use. We encourage participation by underrepresented scientists through the establishment of a NIDA Minority Investigator Award and enhancement of a junior investigator mentoring program. A slate of distinguished plenary speakers, discussion of cutting- edge health services research findings, the development of collaborative relationships, and support of the careers of junior and minority investigators is proposed to move forward an agenda of broadening the positive impact of prevention and treatment services for substance misuse and use disorders.

NIH Research Projects · FY 2025 · 2023-05

PROJECT SUMMARY / ABSTRACT The goals of this proposal are to: 1) obtain experimental skills and career training necessary to develop an independent research program investigating mechanisms of bone development and disease, and 2) characterize a novel target of bone remodeling called G protein-gated inwardly-rectifying K+ channel 3 (Girk3), a key regulator of potassium flux and physiological processes. Our data demonstrate that Girk3-/- mice develop high bone mass after skeletal maturity and have low interleukin-1 beta (IL-1β) and other circulating cytokines. The overall objective of this proposal is to test the hypothesis that Girk3 deletion enhances bone density in adult skeletons by altering the secretion of IL-1β and other monocyte-derived cytokines that modify bone resorption. During the mentored K99 phase, the specific aims are to identify how deletion of Girk3 in monocytes affects bone mass (Aim 1) and to determine how Girk3 regulates cytokine production by blood and bone marrow monocytes (Aim 2). Methods used to evaluate the role of Girk3 in bone remodeling will include dynamic and static histomorphometry, osteoblast and osteoclast differentiation assays, cytometry by time-of-flight (CyTOF), single cell RNA-sequencing (scRNA-seq), and bioinformatics. Completion of this aim will facilitate my transition into the independent R00 phase when the specific aim (Aim 3) will be to determine if Girk3 deletion can prevent bone loss in a murine model of osteoporosis via inhibition of IL-1β. The K99 phase will be conducted at Mayo Clinic and will focus on obtaining mentored training in professional development through meetings with a curated mentorship team, regular attendance and presentations at research seminars and national research conferences, participating in workshops on grantsmanship and responsible conduct in research, and seeking out networking opportunities, as well as conducting the experiments in Aims 1 and 2 and publishing the results. The R00 phase will be conducted in my independent laboratory and will focus on completion and publication of Aim 3 and developing an R01 application based on the results. The proposed plan synergizes new skills in osteoclast and ion channel biology, unbiased spectometry, single cell RNA sequencing and bioinformatics, and bone histomorphometry with prior expertise in endocrinology and osteoblast biology to create my own unique research trajectory. The career development plan and research strategy outlined in this application will produce a robust foundation for an independent research career in musculoskeletal biology.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY Referred pain is both understudied in research and poorly understood in the clinic, particularly for patients with bladder disease. Bladder pain can significantly lessen quality of life, which is amplified by unacknowledged or improperly treated pain from the skin. Diagnosis of referred bladder or somatic pain is obfuscated by a lack of obvious pathology, exacerbating the challenges of finding effective therapeutic approaches. Though the root cause of such referred pain is unknown, it likely relies on visceral and skin afferent interactions, termed viscerosomatic crosstalk. Mechanisms of referred pain attributed to the spinal cord fail to explain why patients with pelvic pain have sensory innervation loss of the lower limb skin that is diagnostic for neuropathic pain. Preliminary data shows that mechanical hypersensitivity in hind paws of mice with bladder inflammation closely resembles nerve injury phenotypes and reflect patient experiences of lower limb sensitivity from bladder inflammation or nerve damage. In the peripheral nervous system, dorsal root ganglia (DRG) neurons are widely diverse in function and in innervating tissue, where injured and uninjured neurons, and their surrounding satellite glia, undergo changes after injury that drive pain. Viscerosomatic crosstalk between uninjured bladder or somatic neurons in DRG could be causing referred pain, but there is a dearth of information about bladder neuron crosstalk in the DRG. Retrograde neuronal tracing studies confirming DRG can co-housing both bladder and hind paw skin sensory neurons strongly support this possibility. The proposed research will test the hypothesis that viscerosomatic crosstalk in DRG after injury results in hyperexcitable physiological responses of the uninjured circuit, mediated by functional changes in their sensory neurons and altered signaling with satellite glia. To do this, neurophysiology experiments will utilize intact DRGs to maintain local communication between neurons and satellite glia, including a novel ex vivo preparation that leaves the mouse sensory circuit from the hind paw skin to the spinal cord intact. Together with molecular assays of protein expression, experiments will determine the mechanisms driving activation of tracer-labeled uninjured neurons by probing activation of satellite glia and two key membrane receptors, transient potential channel V member 1 (TRPV1), also known as the capsaicin receptor, and Purinocepter 3 (P2X3), a widely studied adenosine triphosphate receptor, both of which are poorly understood in referred pain that results from bladder inflammation or nerve injury. Aim 1 is designed to investigate how uninjured hind paw neurons are physiologically altered after acrolein-induced cystitis, and the possibility that these alterations are mediated by changes in TRPV1 or P2X3. Aim 2 will explore how uninjured bladder sensory neurons are affected by Spared Nerve Injury, a robust model of lower limb neuropathic pain. Collectively, these data will help elucidate sensory neuron crosstalk in DRG as a new biological mechanism underlying referred pain in patients with bladder disease and provide a starting point for improved diagnosis and novel, effective therapeutic approaches.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY / ABSTRACT Older Americans are a growing segment of the population with an increasing need for surgical services, and they suffer a disproportionate burden of postoperative complications compared to their younger counterparts. There are opportunities to reduce risk and improve surgical care delivery for this vulnerable population. Preoperative comprehensive geriatric assessment (pCGA) optimizes multiple chronic conditions and factors commonly overlooked in routine preoperative planning, including physical function, polypharmacy, nutrition, cognition, mental health, and social/environmental support; pCGA has been shown to decrease postoperative morbidity, mortality and length of stay in a variety of surgical specialties. Although national guidelines recommend the use of pCGA, a paucity of strategic guidance for implementation limits its uptake to a few academic centers. Systems-engineering and implementation science offer unique tools to reliably improve effective use of this important intervention. The goal of this proposal is to use systems-based engineering methods to tailor and pilot test a user-centered implementation package for the pCGA – one that can be adapted to community-based hospitals in preparation for a multi-site implementation trial. This work will promote equitable access to the pCGA, preserving effectiveness, protecting healthcare resources, and preventing avoidable morbidity and mortality. This goal aligns with the National Institute on Aging’s strategic vision C3 to “Support the development of behavioral interventions based on principles of basic behavioral and social science and designed with an eye to real-world implementation, in line with the NIH Stage Model.” My long-term goal is to improve surgical care for older adults through transdisciplinary research that promotes implementation and dissemination of effective, evidence-based, patient-oriented interventions. This 5-year K23 proposal will provide me with mentored research experience and formal training in implementation science, systems-engineering, clinical trials and aging research. With my strong background in surgical health services research, I am well positioned to conduct this project and transition to research independence, with the support of a team of expert mentors at an institution with an outstanding research environment. The proposed research aims are to: 1) Map the pCGA process and identify system-based barriers and facilitators to its use among older adults undergoing major abdominal surgery 2) Co-design an implementation package for the pCGA applicable to a diverse population of older patients undergoing major abdominal surgery at (2a) a large academic hospital and (2b) an affiliate community site, and 3) Test and refine the pCGA implementation package in preparation for a future randomized controlled implementation-effectiveness trial. Upon completion of this project, I will have an adaptable, user-centered implementation package for pCGA with preliminary pilot data on implementation and effectiveness, which will serve as the basis for an R01 proposal to conduct a multi- site hybrid implementation-effectiveness clinical trial of the pCGA in older adults undergoing abdominal surgery.

NIH Research Projects · FY 2026 · 2023-04

ABSTRACT The BBB acts as a signaling and transport interface between the blood and brain, and with its very low permeability and a wealth of molecular transport systems, the BBB helps regulate the extracellular composition of the brain. While brain microvascular endothelial cells (BMECs) are possessive of these BBB functions, the BBB is greatly influenced by interactions with supporting cells of the neurovascular unit (NVU) such as astrocytes, pericytes and neurons. Recent studies have indicated the importance of CNS pericytes in BBB formation and maintenance, with pericytes triggering reduced transcytosis, reduced expression of leukocyte adhesion molecules and proper tight junction organization in BMECs. Loss of pericyte-endothelial cell interactions and BBB dysfunction are thought to be critical for the pathogenesis of Alzheimer’s disease. Despite the potential importance, the molecular mechanisms driving brain pericyte regulation of the BBB in health and Alzheimer’s disease are largely unknown, particularly in humans. In this proposal, we aim to further examine the mechanisms by which brain pericytes are specified and subsequently impact BBB function. A powerful and innovative approach to explore human pericyte development and function is the use of human pluripotent stem cell (hPSC) technology to model brain pericytes. However, current strategies for differentiating brain pericytes result in cells that lack key brain pericyte hallmarks. We have devised a protocol where brain pericytes can be differentiated from hPSCs by activation of Notch3 signaling in neural crest, yielding improved facsimiles of in vivo brain pericytes. Here, we will further explore the impact of Notch3 signaling on pericyte development and the subsequent effects on BBB induction and maintenance in health and disease. The impact of Notch3 activation in hPSC-derived neural crest will be evaluated by assessing brain pericyte fate, pericyte functionality and the ability to induce BBB properties in co-cultured BMECs. In parallel, using genomics approaches, we have identified a transcriptional network directly regulated by Notch3 activation in pericytes, and have found that this network is downregulated in brain pericytes of Alzheimer’s disease patients. To elucidate the mechanism by which Notch3 activation drives pericyte specification and development, we will use complementary tools of CRISPR-edited hPSC lines and developmental mouse models to systematically regulate the Notch3 transcriptional network and determine impacts on pericyte development and BBB formation and maintenance. Finally, we will assess whether upregulation of Notch signaling in a mouse model of Alzheimer’s disease can ameliorate the effects of pericyte dysfunction on BBB pathophysiology associated with Alzheimer’s disease. Taken together, understanding the impact of Notch3 signaling on pericyte development and BBB function could yield many new mechanistic insights about human BBB induction and maintenance and open new avenues for restoring BBB function in Alzheimer’s disease.

NIH Research Projects · FY 2025 · 2023-04

PROJECT SUMMARY The number of people over age 65 has more than tripled over the past century, placing great burdens on families and the healthcare system. Extending healthspan, the portion of the life that is relatively free from major deficits that impair quality of life, may be the most efficacious way to prevent or delay aging and age- related diseases to reduce such burdens. However, without fully understanding the mechanisms driving aging, identifying and developing effective and translatable interventions to improve health and longevity in humans will likely remain limited. A hallmark of aging, cellular senescence (CS), is an apoptotic-resistant, cell-cycle arrested state induced by various stressors. Senescent cells release pro-inflammatory soluble factors collectively called the senescence- associated secretory phenotype (SASP), which can induce premature CS in healthy cells and promote age-related pathologies. CS can be cleared by genetic or pharmacological (i.e., senolytic drugs) methods, which leads to improvements in metabolic health and reductions age-related diseases. Protein restriction (PR) improves many aspects of metabolic health in both humans and mice, and extends mouse lifespan, specifically through the reduced consumption of the branched-chain amino acids (i.e. leucine, isoleucine and valine; BCAAs). In preliminary studies, I have found that a diet low in BCAAs (BCAA-R) reduces CS in aged and diet-induced obese mice as well as cell culture. The mechanisms by which protein promotes CS, and conversely the mechanisms by which PR and BCAA-R promote healthy aging and reduce CS, are unknown. One mechanism by which BCAA-R may function to reduce CS is the protein kinase mTORC1, which is stimulated by protein and BCAAs. mTORC1 activity promotes the SASP, while the mTOR inhibitor rapamycin inhibits the SASP. BCAA-R has been shown to reduce mTORC1 signaling in the liver and muscle of male mice. Another factor required for PR is fibroblast growth factor 21 (FGF21), which is also induced by BCAA-R. In vitro, FGF21 has been shown to protect against CS burden. In this proposal, I will test if diet affects CS clearance by assessing if senolytics will have the greatest effect on metabolic health and frailty in aged mice eating a high protein diet. I also will test if BCAAs drive high-protein-induced CS and metabolic dysfunction through activation of mTORC1 in vivo and in vitro. Finally, I will test if the action of BCAA-R on FGF21 in the whole body or in specific tissues mediates the effects of a BCAA-R on metabolic health and CS in diet-induced obese mice. Together, these aims will examine multiple mechanisms by which dietary protein and the BCAAs modulate to impact CS burden. I will be completing this fellowship under the mentorship of my sponsor, Dr. Dudley Lamming, an expert in aging biology and metabolism, and my co-sponsor, Dr. Paul Robbins, a world-leading expert in CS. Completing these aims and the accompanying individualized training plan will help me develop the skills necessary to become a successful tenure track faculty member focused on the biology of aging and cellular senescence.

NIH Research Projects · FY 2025 · 2023-04

ABSTRACT Lower urinary tract symptoms (LUTS) are a significant burden to aging men and are often as a result of benign prostatic hyperplasia (BPH). While BPH/LUTS is not commonly fatal with proper medical intervention, it does cause a significant reduction in quality of life for many men as they age. Furthermore, BPH/LUTS increases risk of mortality and results in billions of dollars in healthcare costs annually. There is currently a subset of BPH/LUTS patients that fail FDA-approved treatments (α-blockers, 5ARI), and these patients have been shown to have increased prostatic fibrosis. There is no FDA-approved medication targeting fibrosis or the aging process in BPH/LUTS, even though aging is the greatest risk factor. This proposal aims to develop a better understanding about the role of aging, fibrosis, and mitochondrial dysfunction in BPH/LUTS. Mitochondrial dysfunction is a hallmark of both aging and fibrosis and has not been thoroughly studied in relation to BPH/LUTS. Preliminary data suggests that oxidative phosphorylation (OXPHOS) is a mitochondrial pathway contributing to cellular dysfunction in BPH/LUTS. Aim 1 of this proposal intends to investigate the connection between OXPHOS disruption and lower urinary tract dysfunction (LUTD) in a novel mouse model. Aim 2 of this proposal will investigate the connection between OXPHOS disruption and pathways associated with fibrosis, using genetic loss-of-function cell line models. Finally, Aim 3 will work to identify a new pathway of interest for treatment, using oleic acid as a mitochondrial metabolism modulator. Collectively, these aims will improve our overall understanding of the processes underlying BPH/LUTS, with a special focus on aging, mitochondrial dysfunction, and fibrosis. This proposal hopes to provide translational outcomes that can eventually improve patient care and treatment options.

NIH Research Projects · FY 2026 · 2023-04

Project Summary/Abstract Children’s early vocabulary skills cast a powerful predictive spell over their subsequent language48,96,97 and educational36,83,143 outcomes. At the same time, bilingual children’s language-specific vocabulary skills lag behind those of their same-age monolingual peers14,15,49,58,62,87,88,93,97,146,156,160,161,162,164. This lag in language-specific vocabulary is seen as inevitable and persistent because of distributed exposure of bilingual input across two languages109,113. The crucial gap in the literature is that the effect of distributed exposure on bilingual children’s word-learning has not been tested experimentally. The overarching goal of the proposed work therefore is to test how distributed exposure affects word learning in bilingual children. To this end, we situate the proposed study within the Cross-Situational Statistical Word Learning (CSWL) framework, where the formation of links between words and their referents relies on the ability to aggregate statistical information across multiple exposures to ambiguous mappings between multiple words and referents46,142. Rather than framing our inquiry in terms of bilingual / monolingual comparisons, we instead harness the CSWL paradigm, and focus on experimentally manipulating the exposure parameters within the bilingual participants. Our participants are Spanish-English bilingual toddlers with typical language and who are late to talk, as well as Spanish-English bilingual children with Developmental Language Disorder (DLD). Specific Aim 1 is to test the effect of distributed exposure on CSWL in samples of 2-year-old bilingual toddlers enriched for history of late-talking. In a series of four experiments, we will systematically manipulate exposure parameters, and test specific, mechanistic hypotheses regarding the effects of these manipulations on children’s CSWL. Specific Aim 2 is to test the developmental trajectory of distributed-exposure effects and language outcomes in bilingual children with a history of late-talking. The same children recruited under Specific Aim 1 will be tested again at age 4, enabling us to examine how learning from distributed exposure changes over time, and identify factors that predict recovery of expressive language skills in bilingual late-talkers. Specific Aim 3 is to test the effect of distributed exposure on word learning in 4-year-old bilingual children with DLD. The prediction stemming from bilingual/monolingual comparisons literature is that distributed exposure will interfere with learning (vs. single- language exposure). The alternative prediction stemming from theories of CSWL172,174 and from the limited experimental studies of bilingual learning73,119 is that effects of distributed exposure may be limited, or even facilitative for bilingual children. Proposed studies will move theories of bilingualism and of CSWL forward by identifying the specific mechanisms by which distributed exposure does (or do not) impact word learning in bilingual children. In practical terms, the proposed studies will reveal whether there is an optimal exposure/intervention strategy for successful acquisition of two languages, and whether it works equally well for children with typical language skills and for children who are at risk for language impairment.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Alzheimer’s disease (AD), a progressive and debilitating neurological disorder of old age, is clinically hallmarked by memory loss and neuropathologically by accumulation of Aβ plaques and neurofibrillary tangles in the brain. Synaptic dysfunction has recently emerged as another early key feature of AD; evidence suggests that synaptic density decreases up to 30% in preclinical stages of AD and correlates more closely with cognitive deficits than Aβ pathology. Although age is the single biggest risk factor for developing AD, the observation that even individuals at genetic risk for AD or harboring AD neuropathology are able to remain cognitively normal as they age has refocused research away from risk and underscored the need for investigations of the factors that confer resilience in hopes of reducing disability and disease incidence. KLOTHO is dubbed an anti-aging and longevity gene, and plays a key role in cellular metabolism, central nervous system maturation, and synaptic plasticity. Critical to this proposal is that KLOTHO also seems to enhance synaptic integrity and protects from neurodegeneration. Thus, this integrative, clinically relevant project will rigorously investigate whether KLOTHO 1) confers resilience against age- and AD-related synaptic dysfunction, and 2) modifies the relationship between such dysfunction and cognitive decline both cross-sectionally and longitudinally. The proposed study will be embedded within the robust framework of two well-characterized and longitudinally followed cohorts of ~2,000 at-risk, late-middle-aged adults (the Wisconsin Registry for Alzheimer’s Prevention [WRAP] and the Wisconsin Alzheimer’s Disease Research Center [WADRC]). All participants are already genotyped for KLOTHO and have been cognitively phenotyped for up to 20 years under WRAP/WADRC. The R01 will provide resources for a subset of these participants (N=150) to undergo multimodal biomarker assessment ([11C]UCB-J PET imaging, CSF and blood-based biomarkers) at baseline and 2-year follow-up. Completion of this study has the potential to provide invaluable insights and druggable targets for forestalling brain/cognitive deterioration with advancing age or AD pathological burden.

NIH Research Projects · FY 2026 · 2023-04

We discovered that Nε-lysine acetylation occurs in the lumen of the endoplasmic reticulum (ER) in 2007. From that initial finding, we went on to discover the entire ER acetylation machinery (one membrane transporter, AT-1/SLC33A1, and two acetyltranferases, ATase1 and ATase2) and uncover a novel piece of ER biology. Specifically, we discovered that the ER acetylation machinery regulates proteostasis within the secretory pathway as well as metabolic crosstalk between different intracellular organelles and compartments. Human-based studies discovered that dysfunctional ER acetylation, as caused by loss-of-function homozygous and heterozygous mutations or gene duplication events, is associated with different human diseases, from developmental delay of the brain and premature death to peripheral forms of neuropathy, autism spectrum disorder, intellectual disability and segmental progeria. Mouse models that mimic these genetic events recapitulate associated human diseases. Importantly, ATase-targeting compounds that restore the proteostatic functions of the ER rescue the disease phenotypes of the animals. In conclusion, we have identified a novel molecular machinery that is key to the maintenance of proteostasis within the secretory pathway, and that can be targeted to (i) understand the pathophysiology of several related neurological diseases and (ii) develop appropriate translational approaches to resolve proteostatic defects. The GENERAL HYPOTHESIS of this research is that ATase1 and ATase2 act downstream of an intracellular communication network that regulates the proteostatic functions of the ER and secretory pathway. Our main goal is to dissect the molecular mechanism(s) underlying the acetyl-CoA:lysine acetyltransferase activity of the ATases and understand how ER-based acetylation regulates the efficiency of the secretory pathway. Aim 1 will test the hypothesis that the ATases have divergent functions and differentially regulate proteostasis and metabolic crosstalk. Aim 2 will test the hypothesis that unique structural features allow the ATases to respond to acetyl-CoA influx, Ca++ levels, and perturbations in ER proteostasis. Aim 3 will test the hypothesis that the acetyl group added in the ER lumen by the ATases must be removed in the lumen of the Golgi apparatus by Amfion/GDAC to ensure correct trafficking of nascent glycoproteins and the quality of the secretome. In conclusion, this proposal is based on novel findings from our laboratory and offers a series of highly mechanistic studies that have the potential to define new avenues of research (and treatment) for different neurological diseases. The proposal will use unique mouse models as well as highly integrated novel experimental approaches. We believe that upon completion of these studies, we will have defined an entirely new avenue of research for different neurological diseases.

Fonds de recherche du Québec – Nature et technologies · FY 2023-2024 · 2023-04

Volet: Bourses de recherche postdoctorale; Domaine: Structures abstraites; Objet: R&D et innovation; Objet: Assurances; Application: Sciences et technologies; Application: Fondements et avancement des connaissances; Mots-clés: ACTUARIAT, STATISTIQUE DE L'ASSURANCE, TARIFICATION EN ASSURANCE DOMMAGE, DEPENDANCE, COPULES, MODELES ADDITIFS GENERALISES