Case Western Reserve University

NIH Research Projects · FY 2025 · 2023-05

PROJECT SUMMARY Toxoplasma gondii is an obligate intracellular parasite and causative infectious agent of ocular toxoplasmosis (OT). This chronic, recurrent infection is the top cause of infectious retinitis worldwide and can lead to blindness in one eye in 25% of patients. Current treatment does not positively influence visual outcomes. T. gondii resides in a parasitophorous vacuole and induces mechanisms that block autophagosome-lysosome formation from targeting the parasite in infected cells. After parasite invasion, T. gondii activates the host cell signaling molecule, Src, which drives prolonged autophosphorylation of the EGFR and activates the downstream autophagy inhibitor, Akt. Activated Akt persistently avoids autophagic targeting and survival within the cell. Previously, EGFR inhibition was shown to induce autophagic killing of T. gondii in approximately half of the cells and is partially protective against OT. This partial protection may be a result of partial Akt inhibition and the restricted expression of EGFR. Thus, Src is likely a better target because it appears to activate Akt independently of EGFR and is broadly and highly expressed in neural tissue and the retina. This project seeks to understand the host signaling mechanism utilized by T. gondii in cells lacking EGFR that prevent autophagy-mediated targeting of the parasite, to understand the molecular events triggered by Src inhibition that are responsible for selective targeting of T. gondii by autophagy, and to determine whether Src inhibition protects against OT. Our preliminary studies demonstrate that knockdown of Src induces parasite killing in cells lacking EGFR through an autophagy-mediated mechanism. Additionally, Src inhibition induces autophagic targeting that appears dependent on the activation of a protein kinase. Therefore, the central hypothesis of this proposal is that, even in the absence of EGFR, Src inhibition kills T. gondii due to protein kinase-dependent selective targeting by autophagosomes, and Src inhibition controls OT. Experiments proposed in Aim 1 will investigate the role of Src in preventing the autophagic killing of T. gondii in the absence of EGFR in vitro. Experiments proposed in Aim 2 will explore the role of protein kinase activation in selective parasite targeting by autophagosomes after Src inhibition. Aim 3 will explore the effects of Src inhibition on pre-established OT in vivo. Together these data will define mechanisms in which T. gondii inhibits autophagic targeting, explain how autophagy selectively targets the parasite and may be applied to improved treatment for OT.

NIH Research Projects · FY 2026 · 2023-05

PROJECT SUMMARY / ABSTRACT Hemiparesis of the upper-limb is one of the most serious impairments resulting from stroke. Paresis of finger and thumb extensors is a frequently persisting consequence of stroke, and causes loss of hand function. We have developed contralaterally controlled functional electrical stimulation (CCFES), a novel neuromuscular electrical stimulation (NMES) therapy that gives the patient intimate control of both timing and intensity of stimulation to their finger and thumb extensors and thereby enables intention-driven hand opening and enhanced functional task practice. Several clinical trials of CCFES-assisted therapy have shown that it reduces impairment and improves function of the affected upper-limb, and it improves dexterity more than conventional NMES. The main objective of this study is to build upon the benefits of CCFES-assisted therapy for chronic stroke motor recovery. One strategy to improve rehabilitation outcomes is to combine treatments that may have synergistic effects. Therefore, this study applies transcranial direct current stimulation (tDCS) to the motor cortex during CCFES to determine if the combination of the two will improve outcomes over those achieved by CCFES alone. TDCS and CCFES may work in synergy to improve outcomes by increasing the concurrent activity of the cortical neurons within the ipsilesional motor network (conventional tDCS montage) or by exciting the contralesional networks (unconventional tDCS montage), as suggested by our pilot single-session cross-over study. The specific aims of the study are: 1) Determine if the addition of tDCS during CCFES improves motor outcomes over CCFES alone, 2) Estimate the relative effects of two tDCS electrode arrangements on motor outcomes, and 3) Estimate the relative effects of two tDCS electrode arrangements on neurophysiologic outcomes. We will conduct a randomized controlled trial in which 63 stroke survivors 6 to 24 months post-stroke will be randomly assigned to 12 weeks of: a) conventional tDCS during CCFES, b) unconventional tDCS during CCFES, or c) sham tDCS during CCFES. Upper extremity impairment, activity limitation and neurophysiologic assessment will be made at baseline, 6, 12, 24, and 36 weeks. This study is the first RCT of tDCS during CCFES in chronic upper extremity hemiplegia. The information learned in this study will serve to accelerate the development of treatments for reducing post-stroke disability.

NIH Research Projects · FY 2026 · 2023-04

Prions are the causative agents of a group of transmissible degenerative brain diseases that are fatal to humans and animals. The properties of the infectious agent and its mechanism of replication are unique. Prions are comprised of PrPSc, an abnormally folded, aggregated version of its host-encoded counterpart protein (PrPC); PrPSc self-replicates in the absence of informational nucleic acids by imposing its infectious conformation on PrPC. Our poor understanding of the structural properties of PrPSc represents an important knowledge gap in prion biology. This information is critical to elucidate the detailed mechanism of prion replication, the molecular basis of prion strain variation, and the factors controlling prion transmission between species. In this project, we focus on ascertaining the structural properties of prions causing chronic wasting disease (CWD), an uncontrollable, contagious epidemic of North American deer, elk, and other members of the cervid family, which is also emergent in Asian and European countries. The significant zoonotic potential and emergence of novel strains have established CWD as a significant public health concern. The specific objectives of our research are therefore (i) To determine structural properties of North American elk and deer CWD prions; (ii) To determine the effects of primary structural variation at prion protein residue 226 and route of transmission on CWD prion strain properties; and (iii) To determine the structural basis of strain variation in emerging Scandinavian CWD strains and an adapted derivative. Our proposed studies are truly interdisciplinary since they combine the complementary resources of two laboratories with expertise in novel mouse models for studying CWD, cell culture approaches, biochemical assays, and a host of state-of-the-art methods of structural biology. The later include several complementary mass spectrometry-based structural methods and cryo-electron microscopy, an emerging approach with high potential for providing long-sought insights into high-resolution structures of different prion strains.

NIH Research Projects · FY 2024 · 2023-04

ABSTRACT The Innovative Mentoring and Professional Advancement through Cultural Training (IMPACT) program is a multi-dimensional, inter-institutional, collaborative mentoring program for students who identify with historically underrepresented groups and who are interested in communication sciences and disorders (CSD). This program is a collaboration between Case Western Reserve University, a top US research university, and Hampton University, a top-ranked Historically Black College and University (HBCU). Currently, the program supports 21 undergraduate students in CSD and clinical speech-language pathology graduate students. This supplement has two aims. The first is to further enhance the students’ self-sufficiency and prospects for success by adding group and individual coaching sessions that foster the development of robust professional identities and life skills (including time management, organization, and stress management). This supplement will support a carefully structured approach to cultivating these identities and skills in our current target population. The second aim is to enhance the knowledge, self-efficacy, and sense of belonging of underrepresented Doctor of Audiology (AuD) students. Because AuD programs tend to be small (an average of 10 students), underrepresented AuD students often find themselves in cohorts where none of their peers share their background. Through this supplement, AuD students will have access to the entire IMPACT curriculum and will participate in clinical case discussions, grand rounds, and journal groups, activities that will help build a sense of community and social belonging. They will also have the opportunity to practice and enrich their academic skills. Both coaching and mentoring will be delivered by program faculty who identify with historically underrepresented groups. The supplement will enable us to assess the benefits of coaching paired with our robust mentoring program for the students we currently serve as well as the effectiveness of our expanded model for an online cohort of AuD students.

NIH Research Projects · FY 2025 · 2023-03

Cardiovascular risk from comprehensive evaluation of the CT calcium score exam Summary Using a comprehensive machine learning analysis of coronary artery calcifications and thoracic fat depots in CT calcium score images, we will predict future major adverse cardiovascular events. Improved characteriza- tion of cardiovascular risk will advance knowledge of cardiometabolic disease phenotypes and support clinical therapeutic decision-making and patient counseling for improved adherence. With improved risk prediction and identification of high-risk phenotypes, there will be an opportunity to guide precision preventive therapies, where guidance is needed given the cost and side effects associated with some of these therapies. The Agat- ston calcium score is the leading predictor of a future major adverse cardiovascular event, better than any oth- er single assessment. Epicardial fat volume and HU values are independent risk factors. We will combine analyses of fat and calcifications in CT calcium score exams in an unprecedented way. For coronary calcifica- tions, pathological observations and preliminary results suggest that examining other features (calcium-omics) can improve prediction as compared to whole-heart Agatston. Our assessments will characterize small, spotty, low-density calcifications, providing a better surrogate of disease vulnerability than Agatston, which is numeri- cally dominated by large, likely stable, calcifications. In addition to fat volumes, we will examine quantitative texture and shape features (fat-omics). Elevated HU values and tissue textures are indicative of fat inflamma- tion. All these observations suggest significant value in a combined fat-omics and calcium-omics analysis. We will use large archives of CT calcium score exams from different sites, including University Hospitals of Cleve- land, which is an institution with the largest no-charge CT calcium scoring program (>13,000 scans per year). These big data repositories provide a unique machine-learning opportunity. Numerous technical innovations are planned, including novel features, data representations, and machine learning approaches. In addition to clinical risk prediction, our CT calcium score analyses will dovetail in the future with many research interests, including the role of genes, metabolomics, co-morbidities (e.g., diabetes and psoriasis), socio-economic status, and cardio-oncology on cardiovascular risk.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY Stroke is currently the leading cause of permanent disability and fifth leading cause of death in the United States. Early and accurate recognition reduces mortality and morbidity by expediting access to neurocritical care. Unfortunately, recognition of stroke during triage is dependent on symptom-based assessments that are often unreliable, and up to 35% of patients are misdiagnosed at initial clinician contact. Thus, the development of accurate biomarker-based screening tools that could be used to rapidly detect stroke in emergency medicine settings could substantially reduce rates of mistriage, enable earlier access to intervention, and improve patient outcomes. Numerous prior investigations have attempted to use blood measures of various proteins released from damaged neural tissue as an indicator of stroke. Unfortunately, it is now known that many of the proteins targeted in these prior studies exhibit a limited degree of enrichment in brain tissue, curtailing their diagnostic specificity. Furthermore, they typically circulate at too low of levels to enable reliable detection using the immunoassay platforms available for rapid blood testing in emergency medicine settings, blocking their path to clinical use. Circular RNAs (circRNAs) are a family of circularly continuous non-coding transcripts that are resistant to RNase degradation; they accumulate in long-lived cells such as neurons, and recent work has demonstrated that there are variants which are truly found only in the brain. In addition to the potential for improved diagnostic specificity, targeting brain-specific circRNAs released from damaged neural tissue could have significant translational advantages over targeting the proteins studied for blood-based stroke recognition in the past. The amplification methods used to measure nucleic acids are thousands of times more sensitive than the immunoassay techniques used to measure proteins, which could dramatically increase the odds of detection. Furthermore, because these circRNAs would be cell-free, they could be directly assayed in serum without upstream RNA isolation; this could allow for rapid direct detection using qRT-PCR on existing hardware found in clinical labs, or even methods such as loop-mediated isothermal amplification (LAMP) at the point-of-care. Despite these tremendous advantages, brain-originating circRNAs have never been investigated as a potential source of biomarkers for stroke recognition. In the work proposed here, we aim to use digital droplet PCR to evaluate the absolute circulating levels of five brain-specific circRNAs in 500 patients with various cerebrovascular and non-cerebrovascular pathologies who present with stroke-like symptoms at hospital admission, as well as develop a set of clinically viable custom rapid qRT-PCR and LAMP assays targeting said circRNAs and evaluate their sensitivity and specificity for stroke. This novel, innovative, and highly translational project addresses an area of dire clinical need; we fully expect the assays generated as part of this work to establish the immediate technical and molecular foundation for the downstream development of a pair of rapid stroke triage diagnostics: one that could be deployed for stat testing in the clinical laboratory on existing hardware, and another that could be deployed at the point-of-care.

NIH Research Projects · FY 2026 · 2023-03

PROJECT ABSTRACT The hippocampus is vital for spatial navigation, learning and memory, and is extensively interconnected with many cortical and subcortical regions. In addition to the well-defined long-range glutamatergic projections, emerging evidence suggests that a diverse group of GABAergic inhibitory neurons can also send long-range projections to distant areas. This long-range inhibition is ideally positioned to synchronize rhythmic activity and coordinate cell ensemble activity of multiple brain areas to participate in various behavioral tasks. One such example is theta oscillations – a 4-12 Hz rhythmic activity important for memory encoding and retrieval. However, despite the potential importance of the long-range inhibition in coordinating activity between the hippocampus and other brain regions, its cell identity, electrophysiological properties, connectivity, and behavioral roles remain poorly understood. In our preliminary experiments, we performed anterograde tracing to demonstrate that somatostatin-expressing, but not parvalbumin-expressing, inhibitory neurons in the hippocampal CA3 region project to two subcortical areas known as pacemakers for theta oscillations – medial septum-diagonal band of Broca (MS-DB) and supramammillary nucleus (SUM) in the hypothalamus. Ex vivo ChannelRhodopsin2 (ChR2)-assisted patch-clamp recordings further revealed that these long-range inhibitory neurons preferentially inhibit presumptive GABAergic and glutamatergic neurons in MS-DB and GABAergic neurons in SUM, and optogenetic stimulation of somatostatin-expressing axons robustly entrained firing of postsynaptic MS-DB neurons at theta frequencies. We, therefore, hypothesize that somatostatin-expressing inhibitory neurons in CA3 send long-range projection to coordinate cell ensemble activity of CA3, MS-DB and SUM during theta oscillations. In this proposal, we will perform intersectional viral-based neural tracing and ex vivo brain-slice patch-clamp recordings to define anatomical connectivity and cellular and synaptic properties of long-range projecting somatostatin-expressing neurons, and determine their functional connections with both local and remote target regions. Furthermore, we will employ in vivo electrophysiology, closed-loop optogenetic stimulation, and behavior analysis to determine whether these long-range somatostatin-expressing inhibitory neurons play a role in coordinating activity between the hippocampus and MS-DB or SUM in memory formation. As abnormalities in coordination and synchronization between the hippocampus and other brain areas have been implicated in numerous brain disorders, such as Alzheimer’s disease, schizophrenia, and major depression, the proposed studies will not only deepen our understanding of physiological functions of hippocampal long-range inhibition, but also provide a knowledge base for future studies to examine the role of long-range inhibition in these diseases.

NIH Research Projects · FY 2026 · 2023-02

Project Abstract The organization of 3D architecture of mammalian genome is one of the major mysteries in biology. The invention of genome-wide chromosome conformation capture (3C) technologies, or Hi-C, have revealed a hierarchical 3D genome organization, including compartments corresponding to euchromatin and heterochromatin; with higher sequencing depth, Hi-C data further divided the genome into largely invariant topologically associated domains (TADs), which often serve as the boundary for long-range transcriptional regulation. Recently, with ultra-deep sequencing and improved analysis strategies at kilobase-scale resolution, Hi-C can identify chromatin loops within TADs connecting promoters and cis- regulatory elements in an unbiased fashion. Despite these progresses, an unexplored realm is the relationship between DNA sequence and 3D genome features, especially chromatin loops. It is important to elucidate how the transcription machinery translates the genetic variants into variable events at 3D genome levels and leads to transcriptional or physiological alterations. Here we propose to take an initial step solving the genetics of mammalian 3D genome using a panel of F1-hybrid mouse strains. This is enabled by our new Hi-C pipeline named DeepLoop which can sensitively and robustly identify kb- resolution chromatin loops from low-depth allele-resolved Hi-C data. In aim 1, we will generate comprehensive allele-specific maps of transcriptome, epigenome, and kilobase-resolution 3D genome in β-cells from a panel of 7 F1 mice generated from 8 founder strains, which cover nearly the entire spectrum of mouse genetic diversity. In aim 2, we will carry out integrative analyses to reveal the heritable cis- 3D regulatory modules in β-cells. We will also integrate the existing phenotype QTL, eQTL, and functional genomics data into our analysis. In aim 3, we will create a map of conserved 3D regulome between human and mouse β-cells to infer human biology from mouse genetic data. This project will reveal a large part of the genetics of 3D genome in mouse and serve as a launching pad for future human genome research.

NIH Research Projects · FY 2025 · 2023-02

Pericoronary fat: MACE risk from non-contrast CT and the role of iodine perfusion in contrast CT Summary Pericoronary adipose tissue (PCAT) inflammation is an important, emerging concept in coronary artery dis- ease, giving rise to the “outside-in” theory where inflammatory cells within PCAT, delivered by the vasa- vasorum, influence atherosclerosis plaque progression. Using cardiovascular CT images, we will use ad- vanced image processing and AI to better understand pericoronary fat appearance and to predict major ad- verse cardiovascular events (MACE). As cardiovascular disease remains the most common cause of death in the US, improved early detection, disease prediction, and patient management will positively impact health for many individuals. Using cardiac CT imaging (angiography, CCTA; perfusion, CCTP; and calcium score, CTCS) in elegant experiments and analyses, we will elucidate pericoronary fat assessments and create a new, inex- pensive CTCS assessment of pericoronary fat suitable for screening. As the principal pericoronary fat inflam- mation feature in CCTA is elevated HU, we will use CCTP to assess pericoronary fat perfusion and clarify the role of iodine on existing CCTA signatures, including confounds due to varying filling rates with obstructive dis- ease. Using paired images, we will associate CTCS pericoronary fat features to established ones from CCTA. Using appropriate pericoronary fat features from CTCS exams, we will predict major adverse cardiac events (MACE) without the iodine confound and combine with Agatston to get an even better prediction. Large CTCS cohorts enable interesting research studies. For example, using the serial Coronary Artery Risk Development in Young Adults (CARDIA) study, we will determine if pericoronary fat features precede the appearance of cal- cifications, giving credence to the “outside-in” theory. The CTCS exam is inexpensive (≤$99) at many institu- tions. At University Hospitals (UH), our nationally acknowledged free CTCS program currently servicing >13,000 patients/year with an archive of >65,000 cases, will provide an opportunity for big data, machine/deep learning analysis of PCAT. In addition, improved MACE prediction from PCAT plus Agatston will enable multi- ple future studies on health disparities, genes, cardiometabolic risk, co-morbidities (e.g., diabetes and psoria- sis), and cardio-oncology. To accomplish goals, we have assembled a world class team of biomedical engi- neers and physician scientists at University Hospitals/CWRU with deep knowledge of cardiovascular CT imag- ing and the biology of atherosclerosis. This proposal will ultimately advance the field of predictive medicine and propel new ways to pre-emptively detect at-risk patients so that they can be placed on evidence-based thera- pies.

NIH Research Projects · FY 2026 · 2023-02

The overarching goal of the research in the Salz lab is to understand the regulatory logic and molecular mechanisms underlying cell fate choice and commitment. As loss of cell fate leads to developmental disorders, cancer, and infertility, a comprehensive understanding of the many different mechanisms used by cells to establish and secure their chosen fate is essential. Our current work is focused on the female/male fate choice in the Drosophila germline, where we have found that female germ cell identity depends on the permanent repression of testes genes. This proposal builds on our exciting, published work showing that discrete gene- specific blocks of repressive H3K9me3 chromatin silences key regulatory genes normally expressed in the male germline. The repressive H3K9me3 histone modification is best known for its role in constitutive heterochromatin formation and the repression of transposable elements, but recent work in organisms ranging from fission yeast to humans (including our own) reveals that H3K9me3 peaks are also associated with the silencing of protein- coding genes normally expressed in other tissues. Although these emerging studies identify H3K9me3-mediated gene silencing as a conserved and vital strategy for securing cell fate, there is very little information about how H3K9me3 is recruited to protein-coding genes. And there is no information about whether the mechanism governing H3K9me3-mediated gene silencing in one tissue is generalizable to other tissues. These gaps limit our understanding of H3K9me3's role in silencing lineage inappropriate genes in normal development and may therefore impinge on our ability to diagnose and treat disease. The research proposed in this MIRA application will build on our foundational work in the Drosophila germline to fill these gaps in knowledge. Using the powerful genetic approaches and exciting modern technologies available in the fly, we expect our work in female germ cells to accelerate the discovery of new genes and molecular mechanisms relevant to H3K9me3-medidated gene silencing. Our new studies in the larval brain will extend these findings by determining whether somatic cells use an analogous silencing mechanism. When complete, these studies will provide fundamental insights into one of the least understood epigenetic mechanisms governing cell fate choice and commitment.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Multiple diseases, including graft-versus-host disease, transplant rejection, rheumatoid arthritis, and lung fibrosis are known to be driven by pathological activation of T cells. While T cell activation is a key part of many immune responses, this process can become pathological when T cells inaccurately recognize a patient’s own tissues or in the context of tissue transplantation. While immunomodulatory drugs including corticosteroids and cyclosporine are FDA-approved, these agents act on many immune cell types, leading to broad immunosuppression and severe side effects. Past high-throughput screening efforts identified and validated small molecule ‘Selective Inhibitors of T Cell Activation (SITCAs)’ that function in vitro and in vivo without influencing inflammatory responses in other cell types. While these molecules suggested the potential for novel T cell-selective immunomodulatory agents, lack of understanding of their cellular targets prevented further drug discovery efforts. Exportin-1 (XPO1) catalyzes nuclear-to-cytoplasmic transport of hundreds of proteins and also has established roles in regulating the centromere and transcription. The highly toxic natural product Leptomycin was used to establish that blocking XPO1-mediated nuclear export led to cancer cell death, and later efforts led to FDA approval of selinexor, a Selective Inhibitor of Nuclear Export (SINE), for multiple myeloma patients who have failed at least four prior therapies. Our data establish that multiple Selective Inhibitors of T Cell Activation also target XPO1, but with novel pharmacology: these ‘partial antagonists’ inhibit XPO1’s novel role in the T cell activation process but have minimal effects on nuclear export and are substantially less cytotoxic. These data suggest that XPO1 represents a promising new target for blocking pathological T cell activation, and that the novel partial antagonist profile is desirable to avoid on-target cytotoxicity associated with existing XPO1 modulators. This proposal seeks to understand and optimize XPO1 partial antagonists for application in immune- mediated diseases. First, we seek to use structural and functional assays to understand how different small molecules that bind the same site of XPO1 show such divergent effects on cellular phenotypes including nuclear export and cell viability. In Aim 2, we will establish the cellular mechanisms by which XPO1 modulators block T cell activation, with the hypothesis that dissociation from chromatin of XPO1, NFAT transcription factors, and other chromatin factors plays a central role. Finally, we will use medicinal chemistry to optimize the partial antagonist profile and evaluate leading partial antagonists in preclinical models of T cell function, including using human primary T cells and in a mouse model of lung fibrosis in which T cells are known to play a role. Together these studies will extend XPO1 as a therapeutic beyond late-stage cancer patients by optimizing novel partial antagonists of XPO1.

NIH Research Projects · FY 2026 · 2023-01

ABSTRACT Chronic Kidney Disease (CKD) is common among African American (AA) patients. The excess risk for CKD in this population is partially explained by genetic variations in the APOL1 gene (named G1 and G2) that are unique to African ancestral populations. Understanding the molecular basis for the association between genetic variants and the risk for kidney disease is an important goal in biomedical science. The APOL1 gene is unique to humans and a few primates creating limitations to research approaches using conventionally available experimental models. This career development research proposal is designed to train a promising early stage investigator to address these issues by using organotypic kidney models derived from human inducible stem cells (iPSC) in conjunction with state-of-the-art bioinformatics to interrogate human patient data and explore therapeutics. The current proposal is designed to test the hypothesis that APOL1 kidney disease risk variants drive transcriptional differences that can be identified by the integration of glomerular transcriptomes from people with APOL1-associated kidney disease and ex vivo models, to gain insight into APOL1 function in health and disease. Key preliminary data developed by the applicant demonstrates that a variant-dependent APOL1 transcriptional signatures identified in human glomeruli are conserved in mouse models expressing APOL1 variants. The applicant will acquire new skills in two general areas: 1) expertise in the use of novel ex vivo culture models for mechanistic studies, and 2) computational and bioinformatics analysis of large datasets. These skills will be acquired through mentorship, didactics and a pragmatic research program divided into three specific aims: 1) Define APOL1 risk genotype-associated transcriptional phenotypes in podocytes derived from isogenic cell lines homozygous for the G0, G1 and G2 alleles that are anchored to human glomerular gene expression; 2) Anchor APOL1 variants-associated podocyte transcriptional phenotypes to NEPTUNE glomerular gene co-expression modules and clinical outcomes, and identity compounds that could modulate such phenotypes and associated subcellular processes, and 3) Test the chemical perturbagens’ ability to reverse the APOL1 risk genotype transcriptional phenotype and identify kidney disease mechanisms. Completion of this proposal will provide insights into APOL1 biology and function that could be translated to clinical technology by providing early diagnosis and prognosis tools, identification of therapeutic targets and patient derived renal models. Additionally this proposal aims to establish the Principal Investigator (PI) as an independent translational scientist in nephrology. The plan includes mentored training in patient oriented research, extensive wet-lab training in the production of organotypic models from iPSC lines as well as comprehensive coursework in bioinformatics.

NIH Research Projects · FY 2025 · 2023-01

Abstract The clinical utility of MR images is largely as a qualitative tool without in-built standardization, which requires subjective interpretation and time-consuming analysis. Importantly, these qualitative MRI approaches have demonstrated poor tissue characterization, and poor center-to- center reproducibility, greatly limiting their use in clinical trials. Availability of a robust quantitative imaging tool with high tissue discriminability can directly impact clinical care by offering actionable information to end-user clinicians. As an example, availability of accurate tumor infiltration maps in Glioblastomas, a highly aggressive brain tumor, can pave the way for novel multisite clinical trials in personalized radiation therapy and neurosurgery for improved outcomes. None of the current MRI techniques offer this capability in an accurate and reproducible manner. MRF is a quantitative imaging scan that can address the limitations of qualitative MRI by providing reproducible and physiologically meaningful measurements of tissue properties. We have also shown that utilizing the underlying physical/physiological bounds of the quantitative MRF values improves the reproducibility of the image analysis techniques. Integration of MRF and advanced quantitative analytics could fundamentally address the well-recognized low-reproducibility in qualitative MRI approaches and allow broad clinical translation. In this proposal, we have established an academic-industrial partnership among MRF developers (CWRU), image analysis and AI experts (UPenn), Brain tumor imaging experts (UHCMC), and leading healthcare company (Siemens) to ensure successful clinical translation of the MRF-QIA into the clinical workflow. We will achieve our goal with the following aims: Aim 1: Establish a high throughput MRF scan and assess multisite performance for FDA approval; Aim 2: Fully integrate the MRF-QIA image analytics software into the clinical system for brain tumor analysis; Aim 3: Clinical validation of the MRF-QIA application for infiltration prediction in Glioblastoma patients. This project will add new capabilities to the clinical flow directly impacting the end-user experience and patient care: 1) FDA approval of MRF product scan will allow any Siemens clinical site to add it to their routine patient scans. 2) The MRF-QIA software will be distributed globally through Siemens Global Digital Market and will be available for broad clinical and multisite research applications. 3) The specialized application for GB infiltration prediction will lead to new clinical trials for planning targeted biopsy, extended resections, and personalized radiotherapy by neurosurgeons and neuro-oncologists to eventually provide targeted treatment plans for GB patients.

NIH Research Projects · FY 2026 · 2023-01

PROJECT ABSTRACT The hippocampus is essential for memory formation and spatial navigation. In the classic hippocampal trisynaptic circuit (entorhinal cortex→dentate gyrus→CA3→CA1), dentate gyrus and CA1 act as the primary input and output areas, respectively, that connect the hippocampus with the brain areas outside of the hippocampus. By contrast, CA3 is often viewed as an auto-associative network vital for contextual learning via its intra-hippocampal connections. While CA3’s intra-hippocampal inputs and outputs, including mossy fibers from dentate gyrus, recurrent collaterals from CA3, and Schaffer collaterals to CA1 have been extensively studied, the connections between CA3 and other subcortical areas remain largely unexplored. This proposal aims to fill this critical gap of knowledge by investigating the functional connectivity and behavioral roles of subcortical-to-CA3 inputs. Our preliminary experiments revealed that two subcortical areas outside of the hippocampus – basolateral amygdala and supramammillary nucleus in the hypothalamus – excite and inhibit CA3 pyramidal neuron activity, respectively. We will, therefore, test a central hypothesis that basolateral amygdala and supramammillary nucleus promote and suppress contextual fear learning by enhancing and suppressing the activity of CA3 pyramidal neurons, respectively. In Aims 1 and 2, we will determine whether a selective population of neurons in basolateral amygdala and supramammillary nucleus excite and inhibit CA3 pyramidal neuron activity in vitro and in vivo, respectively. In Aim 3, we will determine the role of basolateral amygdala-CA3 and supramammillary nucleus-CA3 inputs in contextual fear memory. The proposed studies represent a significant shift of focus from traditional research on CA3’s intra-hippocampal connectivity to its extra-hippocampal connectivity. Furthermore, as impairments of CA3, basolateral amygdala, and supramammillary nucleus are strongly associated with many neuropsychiatric disorders, including posttraumatic stress disorders (PTSD), depression, chronic stress and epilepsy, this proposal will provide a valuable knowledge base for future studies to explore the role of these three interconnected regions in disease.

NIH Research Projects · FY 2026 · 2023-01

Project Summary/Abstract Animals have evolved specialized neural circuitry that links sensory input to neuroendocrine and behavioral responses. The proper function of these systems is essential for the health and wellbeing of individuals. Sensory inputs controls some fundamentally important innate behaviors, including mating, aggression and parental behaviors. Dysfunction in these circuits may lead to depression, mood disorders, sexual dysfunction, and aberrant parental behaviors. Here we propose to study the neural circuits that detect and process pheromone information and regulate endocrine and behavioral responses in rodents. In vertebrates, pheromone cues can directly trigger mating rituals and territorial aggression. Many terrestrial species have evolved highly sophisticated vomeronasal systems to detect pheromones. The vomeronasal circuit connects directly to the endocrine systems and influences their output. These circuits are largely genetically determined and there is an intrinsic link between sensory input and the behavioral responses. The mouse vomeronasal circuitry, therefore, serves as an ideal model system to elucidate the neural mechanism of sensory information processing, mechanism of neuroendocrine control and sensory control of innate behaviors. Similar circuits exist in humans but may have been compacted during primate evolution to consist of mostly the main olfactory system, and to include other sensory modalities. The study of the vomeronasal system can provide a roadmap to understand these more complex circuits. The objective of this application is to delineate the vomeronasal circuitry that detects and processes information of two classes of female pheromones. The proposal is based on our study identifying two sets of vomeronasal receptors recognizing pheromones cues that convey the sexual identity and the estrus status of female mice, respectively. In this study, we will determine the contribution of vomeronasal sensory neurons (VSNs) expressing these receptors to sexual behaviors. We will investigate and determine the connectivity diagram between the VSNs and the mitral cells in the accessory olfactory bulb. We will also identify the brain regions and specific cell populations that process information conveyed by these cues and map their connections. These studies are expected to reveal highly specific neural circuits that control mating behaviors

NIH Research Projects · FY 2026 · 2023-01

Project Summary/Abstract Congenital heart defects (CHDs) are among the most common and devastating of birth defects. Abnormal development of cardiac conduction often associates with CHD etiology. Investigating the development of cardiac conduction is essential for us to understand the mechanisms behind such conditions. Development of the cardiac conduction system (CCS) is a complicated transformation that comes about through interplay between molecular signaling, structural properties, and physiological function, including hemodynamics and electrophysiology. While an understanding of the molecular networks has progressed rapidly in recent decades, tools to follow the physiological factors contributing to development and differentiation of the CCS remain deficient. This gap is important to fill. Connecting molecular and functional information is key for understanding pathologies of the conduction system and for guiding potential therapeutic strategies. Optical mapping (OM) of transmembrane voltage or intracellular calcium dynamics in the heart using voltage- or calcium-sensitive fluorescent dyes is a powerful tool for studying cardiac electrophysiology, and has been adapted for imaging early embryonic hearts with great success. However currently, OM is limited to imaging excised embryonic hearts which are stilled with excitation-contraction-uncoupler drugs. Removing fragile tubular hearts from the structure and hemodynamic load of the embryo, and incubating them in dyes and drugs interferes with the normal physiology and does not allow longitudinal study over stages of development. The goal of this project is to develop technology to enable comprehensive, longitudinal imaging of the electrophysiological function of the living, beating heart of the early avian embryo, cultured under near- physiological conditions. We will target 1-2 days of active morphogenesis through the transition from homogeneous to heterogeneous conduction velocity Three key technology developments are needed to achieve this. (A) Episcopic, volumetric, fast imaging of fluorescent voltage and calcium indicators is needed to image the intact, living embryo, and to capture conduction dynamics (Aim 1). (B) Motion correction is needed to enable conduction mapping of the beating heart, without using excitation-contraction-uncoupling drugs (Aim 1). (C) Embryonic quail models with calcium and voltage reporters expressed in cardiomyocytes are needed to enable in vivo and longitudinal imaging of electrophysiology (Aim 2). The proposed technology will enable simultaneous 3D conduction mapping over two time scales, the heartbeat, and heart development (5D impulse mapping). Coupled with quantitative 3D FISH, this will allow point-to-point 3D registration between conduction data and gene/protein expression, which is not currently available, enabling studies to better understand mechanisms of conduction function, dysfunction and development (Aim 3).

NIH Research Projects · FY 2026 · 2022-12

Both inside the central nervous system and outside in the peripheral nervous system, cancer cells grow along nerves as routes of invasion and metastasis called neural invasion. This growth is common in several carcinomas including breast cancer and is associated with poor prognosis. Proteolysis of cell adhesion molecules (CAMs) occurs in development, and growing evidence suggests this post-translational modification may promote tumor migration and invasion on nerves that ultimately leads to metastasis to the brain in various tumor types including breast cancer. The receptor protein tyrosine phosphatase PTPµ is a CAM that is proteolyzed in cancer cells to generate an extracellular fragment that is a unique imaging biomarker of the tumor microenvironment. The PTPµ- targeted agents we developed bind to this biomarker and recognize human brain tumors as well as invasive primary breast cancer and breast cancer that has metastasized to the brain. Systemic delivery of the PTPµ- targeted agent results in binding to tumor cells within minutes in xenograft models. Using a 3D cryo-imaging system we analyzed the extent of cell migration and dispersal within the brain. We found that the PTPµ-targeted agent labels 99% of all dispersing tumor cells far away from the main tumor mass on nerves in mouse models. This proposal represents the convergence of our expertise in neuroscience, cell adhesion, imaging and cancer to test if the PTPµ biomarker can be used to detect tumor growth along nerves leading to brain metastases. Gold nanoparticles (AuNPs) have shown outstanding versatility in biomedical applications including imaging diagnostics, drug delivery, and radiation therapy. In this proposal, we describe the development of theranostic AuNPs for the detection and treatment of breast cancer metastases. We will achieve more sensitive detection and treatment of invasive and metastatic lesions through the use of a three component theranostic nanoparticle containing: 1) a highly specific targeting agent of the PTPµ biomarker in the tumor microenvironment; 2) a protease-sensitive quenched near infrared fluorophore for fluorescent imaging; and 3) a gold nanoparticle (AuNP) for sensitization to radiotherapy. We will test whether the PTPµ-targeted agents detects nerve associated growth using 3D single cell resolution cryo-imaging that precisely tracks migration of individual cancer cells on nerves. We will utilize our established human patient-derived xenograft models of metastatic breast cancer and models that metastasize from the breast to the brain. Metastatic tumors are resistant to almost all chemotherapeutics so “physical” killing strategies like radiation must be improved and employed for better therapeutic outcomes. By delivering PTPµ-targeted conjugated AuNPs directly to primary and metastatic breast cancer we will exploit the radiosensitization of AuNP to reduce the required dose of radiation needed for radiotherapy thereby reducing collateral damage to normal surrounding tissues. We expect that these studies will yield targeted nanoparticles that detect and treat nerve associated tumor growth while implicating CAM proteolysis as a generalizable mechanism for detecting and treating tumor invasion on nerves.

NIH Research Projects · FY 2026 · 2022-12

Project Summary Accumulating evidence indicates that the ability to mount an effective Nrf2-mediated gene expression response to oxidative stress declines during the aging process. In particular, nuclear but not cytoplasmic Nrf2 is depleted in neurons of AD patients. In animal models, loss of Nrf2 signaling exacerbates amyloid and tau deposition, neuroinflammation, and cognitive deficits, whereas induction of Nrf2 signaling protects against these phenotypes. While toxic tau assemblies, oxidative stress, cytoskeletal disruption, and autophagy defects are cardinal features of tauopathies, including AD, how these cellular brain phenotypes integrate at the molecular level to produce physiological or pathological responses during tau pathogenesis is unknown. In addition to the known regulation of the cytoskeleton, mitochondria, and autophagy by SSH1, our new preliminary studies show that the SSH1 pathway intersects with the Nrf2 to inhibit and titrate Nrf2 signaling. Our overarching hypothesis is that the nexus between the SSH1 and Nrf2 pathways represents a tipping point that tips the balance between degeneration and protection during proteotoxic and oxidative stress in tauopathies. As both Nrf2 and SSH1 are activated under oxidative stress, understanding how the nexus between Nrf2 and SSH1 is physiologically and pathologically regulated will provide key insights into treating AD and other tauopathies. Utilizing in vitro recombinant proteins, cellular models, animal models, and postmortem brains combined with mechanistic biochemical, immunochemical, in situ proximity ligation assays, RNA- seq, and proteomics studies, we will define and dissect how the SSH1-Nrf2 nexus tips the balance between neurodegeneration vs. neuroprotection in tauopathies.

NIH Research Projects · FY 2026 · 2022-12

PROJECT DESCRIPTION As a major limitation in contrast enhanced MRI studies is that current MRI methods lack the combination of accuracy, precision, and temporal resolution to quantitatively measure contrast agents in vivo. We have addressed this problem by developing dynamic Magnetic Resonance Fingerprinting (MRF) methods that can rapidly measure T1 or T2 relaxation times with outstanding accuracy and precision (Radiology, 2021). Our key MRF innovations include a combination of highly undersampled spiral trajectories, low flip angles and multiple magnetization preparations to avoid errors from B1 inhomogeneities and limited T2 sensitivity. Our MRF methods can also be adapted to simultaneously detect one or two MRI contrast agents. We have recently demonstrated that a new T1-MRF method can be used to dynamically generate quantitative T1 maps with very fast temporal resolution (~2.5 seconds) during an in vivo Dynamic Contrast Enhanced (DCE) – MRF experiment. In Aim 1, we will first optimize a new 3D T1-MRF method to evaluate tumor vascular perfusion (ktrans) with high accuracy and precision in mouse cancer models. We will then evaluate this Dynamic Contrast Enhanced (DCE) – MRF method by measuring changes in vascular perfusion in mouse cancer models treated with either a vascular disrupting agent or radiotherapy. Our objective is to demonstrate that DCE- MRF provides superior precision in comparison to standard DCE-MRI methods providing the opportunity to more sensitively detect the early response to treatment, which can then be translated to the clinic. We have also demonstrated that a similar dynamic MRF method can be used to simultaneously measure the concentration of a T1 contrast agent and a T2 contrast agent within an in vivo tumor model with outstanding accuracy and precision (Scientific Reports 2017 and 2019). In Aim 2, we will develop a similar two-agent MRF method to simultaneously detect a pH-dependent T1 contrast agent and a pH-independent T2 contrast agent to measure extracellular pH (pHe) in tumor models. We will apply our pHe-MRF approach to monitor changes in tumor pHe after administering treatments that raise and lower tumor acidosis to validate our methodology. Our deliverable for this project is a new adaptable, dynamic 3D MRF approach to quantitative measure one or two MRI contrast agents in vivo. These new 3D DCE-MRF and pHe-MRF methods are the key innovation of our research. We have developed a rigorous research approach with an emphasis on quantitative evaluations and validations using multiple established mouse cancer models and therapeutic strategies. We have also assembled a team of strong and highly experienced investigators, and we have an exceptional research environment for our studies. Importantly, this successful preclinical imaging project will immediately lead to clinical translation of the DCE-MRF method for use in cancer patients and will provide the opportunity for effective pHe assessments in animal models and eventually in patients.

NIH Research Projects · FY 2026 · 2022-11

Candida auris has emerged as a global threat causing serious invasive infections with mortality approaching nearly 60 percent worldwide. The majority of C. auris infections are nosocomial and reportedly resistant to fluconazole (FLU) and amphotericin B (AmB) with variable resistance to members of the three major classes of clinically available antifungals (azoles, polyenes, echinocandins), with some strains resistant to all three antifungal classes, thereby limiting treatment options. C. auris has been classified as a ‘newly emerging threat’ by the Centers for Disease Control (CDC), although Candida albicans remains the most prevalent and pathogenic Candida species. With C. auris classified as a global urgent threat, and Candida spp. and other resistant fungal species emerging, there is a need to identify and develop new modalities to treat infections caused by Candida spp. Because C. auris colonizes the skin and acts as a nidus of infection, developing a drug that can concurrently target skin and exhibit systemic efficacy will be highly innovative and desirable. A triterpenoid class of antifungals derived from enfumafungin, a hemiacetal isolated from fermentation of Hormonema spp. (generically termed fungerps), represents the first newly described class of antifungal compounds since 2001. Currently, a second-generation fungerp, SCY-247, is under development as a potential systemic therapeutic option. Because the skin is a natural niche for C. auris, and transmission occurs through cutaneous contact, demonstrating that SCY-247, in addition to working systemically (including at the blood brain barrier), is effective as a cutaneous (i.e., decolonization) treatment for C. auris is innovative. We hypothesize that our preclinical models of systemic and cutaneous C. auris infection, will allow us to demonstrate the efficacy of SCY-247 as a new therapeutic treatment for skin as well as disseminated infection including brain infection. Thus, our multi-pronged approach includes three specific aims designed to: 1) Determine the effective in vitro and in vivo range of SCY-247 that is inhibitory/fungicidal to Candida strains resistant to traditional antifungal therapy with an emphasis on C. auris strains; and 2) Employ established cutaneous guinea pig and murine models of C. auris to address treatment and decolonization approaches using SCY-247, and finally; 3) Determine the efficacy of SCY-247 in the treatment of central nervous system (CNS) infection caused by C. auris or C. albicans spp. using an intracranial murine model and high-resolution intravital microscopy (IVM). Successful completion of these aims will determine whether SCY-247 is a viable option for eradication of C. auris, and whether this compound is effective against known resistant Candida spp. We will evaluate both oral and I.V. dosing of SCY-247 comparing their potential efficacy. Finally, the ability of SCY-247 to treat C. auris brain infection will be assessed. Successful completion of these preclinical studies will enable advancing SCY-247 into Phase 1 clinical trials.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Currently our understanding of how the nervous system maintains ocular surface homeostasis is extremely limited. New technologies, methods and models are needed to advance our scientific understanding and address knowledge gaps. The ocular surface and tear film-secreting glands (including the lacrimal and meibomian glands, as well as the goblet cells) are carefully controlled to provide an optically smooth, low-scattering surface with appropriate immune and injury responses. Sensory feedback to maintain the structural and functional integrity of the ocular surface is provided by the corneal nerves, which send feedback from stimuli (chemical, thermal, mechanical) to ganglia (e.g., trigeminal) and brain regions (e.g., ventral posteromedial thalamus) to drive production of tear film components as well as the blink reflex. This delicate balance of neural control is disrupted by damage, peripheral neuropathies, inflammation and further complicated by a wide array of immune responses to various diseases. Dysfunction of this feedback loop can lead to a downward spiral of further dysregulation. Aberrant neural control of the ocular surface can lead to abnormal sensation and pain, which in the worst cases can be disabling. To find remedies, it is first essential to understand the underlying neural control system and how it adapts to its environment. In this proposal, we aim to bring new tools and models to study molecular, cellular, and functional interactions across systems responsible for neural control of the ocular surface and examine how they change under different inflammatory and pain conditions. We have assembled an excellent team with expertise across multiple fields including advanced 3D microscopy, neuroscience, electrophysiology, pain, ocular immunology, ocular lipid metabolism, ocular surface disorders, spatial statistics, and machine/deep learning. Here, we will utilize cutting edge techniques and technologies including optical clearing, tract tracing, ethologically-valid behavior analysis, machine/deep learning, spatial statistics, genetically encoded calcium imaging, light-sheet microscopy, multiplexed 3D fluorescence in situ hybridization (FISH) imaging, and multi-array electrodes implanted in the brain. These tools will help us assess molecular, cellular, and functional interactions across organs and begin to understand ocular surface control at the organism level. We will also employ several relevant animal models to assess ocular surface control under different inflammatory and pain conditions. Models include AWAT2 deficient mice that mimic evaporative dry eye disease (DED), diabetic mice, an epithelial debridement model with Pseudomonas aeruginosa that mimics bacterial keratitis, and human donor eyes. The mouse models all have gCaMP6f expressed in corneal nerves allowing functional imaging of calcium transients. With these models we will study both innate and adaptive immunity as well as nociceptive and neuropathic pain responses. In addition, we will apply nerve growth factor (NGF) to our models to study how a potential treatment option alters the ocular surface control system.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Functional rhodopsin (Rho) composed of apoprotein opsin and covalently bound 11-cis-retinal chromophore is required for normal phototransduction and vision. Inherited mutations compromise proper folding, binding of 11-cis-retinal, and stability of Rho, leading to retinitis pigmentosa (RP), a progressive degenerative eye disease that causes blindness. Currently, there are no treatment options available to combat RP. Retinal- based pharmacological chaperones and non-retinal small molecules, including flavonoids, can stabilize certain Rho mutants in vitro. However, effective and safe therapy for RP has yet to be established. Dietary flavonoids have been reported to show beneficial effects in ocular impairments. Our preliminary studies showed that common flavonoid quercetin could stabilize pathogenic Rho, improve its folding, membrane integration, and retinal binding. We also found that quercetin inhibits stress-induced cellular responses related to pathogenic mutants triggering Bax-mediated apoptosis with beneficial consequences to retinal health. Here we propose to study, the therapeutic potential of quercetin and its combinations with retinal analogs, and novel small molecule Bax inhibitor to rescue RP pathology. First, we will evaluate quercetin positive effects on RP mutants independently of the genetic background in vitro. Then, we will assess their ability to revert the RP phenotype in vivo in mouse models of RP. To understand the underlying molecular mechanism of Rho mutant’s stabilization offered by quercetin we will perform structural studies to delineate the architecture of quercetin-Rho complexes by applying crystallographic and hybrid mass-spectrometry techniques. In addition, we will search for more effective compounds stabilizing Rho mutants by high-throughput screening of flavonoid-related compound libraries, which then will be validated with systematic biochemical and biophysical analyses (Aim 1). Second, we will examine the role of quercetin as an adjuvant for the specific retinal analogs stably accommodating the retinal-binding pocket. Some of these retinals improve the folding of the P23H pathogenic Rho mutant. We will examine the potential corrective effect of new locked retinal and backbone- modified retinal analogs for P23H Rho and determine if the binding of these retinals could be enhanced by quercetin to gain a greater therapeutic effect in RP (Aim 2). Third, we will study the cellular processes in RP. To gain a better understanding of RP pathology we will look for molecular targets activated by cellular stress signals in RP and assess if quercetin can modulate these cellular processes to revert the disease phenotype (Aim 3). At first, we will examine the effect of quercetin on Bax-mediated apoptosis to learn if its effect is related to direct or indirect inhibition of Bax, or both. In these studies, we will also use a novel bioavailable small molecule Bax inhibitor. We will determine the effectiveness of this new Bax inhibitor and its combination with quercetin against Rho-related RP. Altogether, the knowledge gained from this study will pave the way to design an effective therapeutic remedy to prevent or slow down RP pathology.

NIH Research Projects · FY 2024 · 2022-09

Respiratory complications account for significant morbidity and mortality in persons with cervical and high thoracic spinal cord injury (SCI) due to their inability to cough. As a consequence, most individuals suffer from a markedly reduced ability to clear airway secretions resulting in the development of recurrent respiratory tract infections.1-16, 128-130,132,135,136 Moreover, diseases of the respiratory system are the leading cause of death in this patient population.3,7,9,12,15 These individuals lack an effective cough mechanism due to paralysis of the muscles of expiration.18-20,42,126-130,132,135,136 We have performed a pilot clinical trial (n=17) with disc electrodes and demonstrated that an effective cough can be restored in SCI.27-29 This method results in near maximal expiratory muscle activation, as reflected by the generation of large airway pressures and peak expiratory airflow rates, which are in the same range as those observed during maximum cough efforts in normal persons.27,28 Use of the system significantly reduces the difficulty in raising secretions and reduces the incidence of respiratory tract infections29,117 Unfortunately, disc electrode placement requires an invasive surgical procedure. In a recent clinical trial of bipolar SCS with parallel wire electrodes (T9–T11 spinal levels), we demonstrated that comparable levels of expiratory muscle activation and clinical benefits can be achieved, when compared to the disc electrodes (n=12).119,120,121a,153 Our current stimulation system entails use of wire electrodes manufactured by Ardiem (Indiana, PA) and electrical stimulator by Finetech, Inc (London, England) (FM). FM however does not have Bluetooth capability, has a cumbersome antenna and does not have a license to allow US commercialization. Therefore, we plan to partner with Avery Biomedical Devices (Commack, NY) (ABD) to construct more advanced and more user-friendly stimulator. The ABD device will have Bluetooth capability allowing for remote activation and facilitate data logging including the ability to view, download and analyze data in real time and the antenna is very flexible with no orientation requirement. In addition, ABD already services patients with respiratory care needs. This stimulator will be used in conjunction with Ardiem wire electrodes, utilized in our recent clinical trial. This device will represent the only commercially available spinal cord stimulator with the range of stimulus parameters required to activate the expiratory muscles and restore an effective cough. The Major Objectives of the proposed study, therefore, are to 1) construct an electrical stimulator to activate the expiratory muscles and connect with the Ardiem electrodes 2) perform the necessary testing required by the FDA to amend our current IDE 3) perform a multi-center clinical trial to determine the utility of SCS with wire leads to produce an effective cough and provide an effective means of clearing airway secretions, reduce the incidence of respiratory tract infections and improve subject quality of life in SCI and 4) meet the necessary regulatory requirements to obtain FDA approval of the device for eventual commercialization. Ultimately, we expect this technique to become an important clinical tool in the routine management of subjects with SCI. 7

NIH Research Projects · FY 2025 · 2022-09

Abstract This EDRN-CVC proposal is aimed at the validation of molecular biomarkers for distinguishing high versus low risk esophageal neoplasias (Barrett’s esophagus) for the purpose of guiding selection and management of patients for endoscopic eradication therapy (EET). Two validation studies are proposed: the first, a phase 4 prospective study to identify a patient group at low progression risk who can be spared EET; the second, a phase 3 retrospective study to distinguish individuals who following EET are at low versus high risk of disease recurrence. Barrett’s esophagus (BE) is the precursor lesion of esophageal adenocarcinoma (EAC), a cancer with 80% lethality whose incidence has increased more than 7-fold in the past three decades. BE progresses to EAC in a step-wise fashion from non-dysplastic BE, to low grade dysplasia (LGD), to high grade dysplasia (HGD), and finally cancer. EAC prevention is based on using EET to ablate HGD BE before it can progress to EAC. However, increasingly, EET is also becoming the default therapy for LGD, a highly imprecise diagnosis about which expert pathologists frequently disagree, and which is applied to as many as 40% of BE patients at some point during their course. As EET has a 9% complication rate, the result is an emerging epidemic of overtreatment of BE with LGD. In a prior EDRN-BDL award, our team developed the “BAD” technology for early detection of BE progression. In BAD, we used a brushing device to sample a patient’s full BE esophageal segment. We then analyzed the DNA from this sample using next-generation sequencing technology (developed for liquid biopsy assays) to instead detect presence of BE clones that had acquired gains or losses on specific driver chromosomes associated with EAC. Detection of driver chromosome changes (dubbed Very-BAD), typified EAC and HGD. In contrast, 28% of LGD showed complete absence of any chromosomally aberrant clones (dubbed Not-BAD). We will now validate Not-BAD as a biomarker that identifies LGD at such low progression risk as to not require EET. We will do this by partnering with the SURVENT trial, that will be the first U.S. prospective study to follow LGD patients managed by surveillance, not ablation. A second major challenge with EET is that over 25% of patients recur following ablation (with either high risk BE, HGD, or EAC). These patients face a substantial burden of post-EET surveillance endoscopies, initially at every 3-month intervals. In our prior EDRN-BDL, our team identified a panel of methylated DNA biomarkers for sensitive molecular early detection of BE (currently awarded FDA breakthrough device designation). We have further identified that these markers remain retained in a subset of patients post-EET. We accordingly now propose a retrospective Phase 3 study to further validate these DNA markers for molecular assessment of minimal residual disease, whose post-EET elimination identifies individuals achieving complete molecular eradication of BE, and hence at low risk of disease recurrence and not in need of intense post-EET surveillance. We do this by partnering with the unique UNC-BEECAB biorepository of post-EET esophageal biopsies from patients whose disease did or did not recur following ablation.

NIH Research Projects · FY 2025 · 2022-09

Socioeconomic disparities in glycemic control and diabetes technology use are widely recognized in patients with type 1 diabetes (T1D). Data shows that children in the US in households with the lowest income, lowest education level and public insurance have higher HbA1c’s, are half as likely to use insulin pumps, and over 3 times less likely to use a continuous glucose monitor (CGM) than those with the highest income, highest education, and private insurance. Automated insulin delivery (AID) systems, which require both a CGM and insulin pump, show greatest HbA1c improvement in those with baseline HbA1c over 8%, leading to concern that low use of AID in patients will further exacerbate existing health-care disparities. Therefore, novel interventions to reduce disparities in technology use among lower-income patients with T1D are critical to improving glycemic control, reducing microvascular complications and preventing early mortality in these children and young adults. We aim to reduce disparities with a 2-part intervention: a diabetes triage dashboard with interactive smartphone application and a community health worker (CHW) in the role of diabetes technology coach. An advisory group of stakeholders will define metrics of triage zones for the dashboard and frequency/content of messages from the application and will refine the specific role of the CHW. Our shared goals are to 1) identify patients who struggle to initiate or continue their device or do not meet glycemic goals and 2) utilize a CHW to act as a diabetes and technology coach, building trust with families so as to identify and overcome barriers to successful technology use. The specific aims are to: 1) Convene an advisory group of stakeholders to develop a protocol for a diabetes dashboard and smartphone application; 2) Develop a diabetes dashboard and interactive application using remote data capture from the cloud from CGM, insulin pumps, and AID systems; 3) Assess effectiveness and implementation of the bundled intervention with a 1:1 randomized controlled study of children and young adults with new onset and existing T1D who are not using AID; 4) Aim 4: Understand lived experience of diabetes management in context with 1:1 interviews, considering both perceptions regarding the intervention itself (vs. usual care) and organizational/secular characteristics influencing that experience.