University Of California Los Angeles

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY / ABSTRACT (OVERALL) Autoimmune endocrinopathies include a number of conditions in which the immune system destroys healthy hormone-producing tissues. These conditions affect a large number of Americans, many of them female, but have not yet been the focus of ACE studies in the past. Moreover, unlike most other autoimmune conditions, autoimmune endocrinopathies are not usually treated with immunotherapies, despite clear evidence that their etiology is immune-mediated. Thus, there is an urgent need to better understand autoimmune pathogenesis to identify targets for immunotherapies in these conditions. This new ACE proposal seeks to delineate mechanisms of autoimmune chronicity as critical pathways in the establishment of autoimmune endocrinopathies. The Principal Project delineates the epigenetic and transcriptional mechanisms underlying chronicity in autoimmune endocrinopathies. These studies will determine whether an epigenetic regulator UTX may a role in conversion of stem-like progenitor CD8+ T cells to effectors in autoimmune endocrinopathies. The Collaborative Project addresses spatial and temporal determinants of immune chronicity in cancer immunotherapy-related immune related adverse events (IRAEs). Endocrine IRAEs of great interest given the large proportion of cancer patients who now receive immunotherapies and develop these unwanted side effects. Finally, many autoimmune endocrinopathies are sex-biased, and the Pilot Project addresses sexual dimorphism in immune chronicity pathways. In particular, UTX is an X-linked gene that is differentially expressed in male vs. female CD8+ T cells, and we will explore UTX’s role in mediating sex differences in progenitor to effector conversion. Our ACE includes an Administrative Core, which will ensure efficient day-to-day operational support; an ACE Funds Management Core, which will administer financial and consortium agreements; and an ACE Biorepository Core, which takes advantage of the well-developed biorepository infrastructure system in place at UCLA. Our program will build a cohesive and multi-disciplinary team of immunologists, clinicians, and computational biologists to contribute our deep expertise to the ACE collaborative enterprise.

NIH Research Projects · FY 2024 · 2024-09

Project Summary Tissue factor pathway inhibitor-2 (TFPI-2) consists of three Kunitz domains arranged in tandem flanked by a short acidic amino terminus and a highly basic C-terminal tail. The N-terminal Kunitz domain (KD)1 of TFPI-2 is the only inhibitory domain and it inhibits plasmin (Pm), factor (F) XIa, plasma kallikrein (pKLK), and very weakly FVIIa/tissue factor. The structural specificity of TFPI-2 KD1 for each protease is essentially unknown. The focus of this proposal is to delineate the TFPI-2 KD1 residues, which impart specificity toward Pm, FXIa and pKLK through structural and functional studies. Another goal is to design and test novel, potent and specific inhibitors of fibrinolysis, which have the potential to reduce bleeding in major surgeries without causing adverse effects attributed to broad spectrum protease inhibitors of Pm, like bovine pancreatic trypsin inhibitor (BPTI, Aprotinin). Using structure-based investigations and analyzing the serine proteases substrate specificity profiles, we have developed two TFPI-2 KD1 variants to specifically inhibit Pm. These TFPI-2 KD1 variants are dual reactive inhibitors that selectively inhibit plasmin active site as well as the activation of plasminogen by binding to the kringle domains of plasminogen/plasmin and tissue plasminogen activator. Our new data indicate that these two TFPI-2 KD1 variants are potent and selective inhibitors of Pm with no detectable inhibition of FXIa and pKLK. We will study the efficacy and toxicity of these two KD1 variants in mouse models of bleeding. Since trauma associated hemorrhage and coagulopathy are leading causes of mortality, we will also study the safety and efficacy of the potent KD1 variant in a rat trauma model, where the most potent KD1 will be selected based on the results from the mouse bleeding models. The three overlapping areas to be investigated in this proposal are: 1) Delineate the residues/segments that impart protease specificity of KD1 for Pm, FXIa and pKLK. 2) Assess the efficacy and safety of the two TFPI-2 KD1 variants in two mouse models of bleeding. 3) Evaluate safety and efficacy of the most potent KD1 variant in a rat liver hemorrhage model in acute and chronic settings. The proposed studies will significantly improve our understanding of the specificity of the proteases involved in coagulation and fibrinolysis. Further, it will lead to the development of a potent, specific inhibitor of Pm that can be used as a therapeutic agent to inhibit fibrinolysis. The applications of such therapeutic agents are broad and encompass trauma, cardiovascular surgery, as well as neurologic and orthopedic operations that are complicated by fibrinolysis.

NIH Research Projects · FY 2024 · 2024-09

Project Summary: There remains a critical blind spot in the study of biological systems. In situ imaging techniques like super-resolution microscopy can elucidate cellular dynamics but cannot provide the sub-nanometer resolution needed for molecular detail. Cryogenic electron microscopy (cryo-EM) does provide atomic-resolution structures yet remains limited to only systems in their equilibrium states. While both these techniques have been awarded separate Nobel prizes for their transformative impact on our understanding of biology, a missing gap still exists for how the structure of biomolecules change in their active state far from equilibrium. The proposed research seeks to bridge this gap. This project will pioneer the development of electrified cryo-EM (eCryo-EM), a novel tool developed by my research group that can kinetically trap biological systems in their metastable state away from equilibrium. Briefly, an electrical stimulus applied just prior to and throughout a plunge-freezing step will capture and preserve metastable states in electrically excitable biological systems. This approach in using eCryo-EM to capture metastable state is general for all of biology. Indeed, many fundamental questions remain unanswered for a broad spectrum of electrically excitable biological systems with important implications for medical applications (e.g., wound healing, electroporation drug delivery, muscle contraction, sensory acquisition, etc.). As a starting point, this proposal will unravel the first snapshots of neuron conformational changes through the stages of an action potential across multiple length scales. At the molecular level, eCryo-EM will reveal the molecular structure of voltage-gated channel proteins as they change through open, closed, and inactivated states. At the cell level, eCryo-EM will reveal the mechanism of neural inhibition at the synaptic cleft under high-frequency waveforms. These new insights not only provide fundamental understanding for neuron behavior under an electrical stimulus, but also will guide future neurostimulation treatments that are directed and tailored for a specific neural circuit to restore human health in patients suffering from neurological disease. 3 tasks are outlined to achieve the stated goals: (1) demonstration of an eCryo-EM device that can successfully trap metastable states of biological systems under an electrical stimulus; (2) elucidating a time series of structural changes in voltage-gated channel proteins throughout an action potential; and (3) trapping the multiple conformations of a synaptic junction between neurons throughout electrical stimulation. These tasks cannot be achieved through conventional methodologies. Rather, it requires a fundamentally new approach in kinetically trapping the active state biological systems directly under an electrical bias. The proposed research will establish a new paradigm in investigating how neurons fundamentally behave in response to an external electrical stimulus, thereby providing new neurostimulation therapeutic targets and strategies for treating neurological diseases.

NIH Research Projects · FY 2025 · 2024-09

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Iron deficiency (ID) and iron deficiency anemia (IDA) are major contributors to the global burden of disease, with ~1.25 billion cases of IDA worldwide. In the United States, ID and IDA are more common in disadvantaged populations, including the low-income groups, Indigenous peoples, and migrants from low-to-middle income countries. The recent realization of the importance of ID as a comorbidity that alters outcomes underscores the need to broaden investigation of the effects of ID in different conditions. We observed in a mouse model an unexpected strong adverse interaction between iron deficiency and inflammation that resulted in increased production of inflammatory mediators and exacerbated apoptosis and tissue injury in the lung, kidney, liver and spleen. This suggests that ID may worsen tissue injury in sepsis, and potentially other inflammatory conditions. Considering the high prevalence of concomitant ID and inflammation around the world, the problem has compelling translational importance. We here propose to define the molecular mechanisms underlying the adverse synergy between ID and inflammatory injury, focusing on macrophage-mediated inflammation, endothelial sensitization to inflammation, and resulting lung and kidney injury. In preliminary work, we uncovered a novel mechanistic link between ID and inflammatory damage whereby ID potentiates inflammation through the NF-κB pathway, and also alters tissue responsiveness to inflammation by modulating the CREB transcription factor and ER stress. We will address the mechanisms in the following aims: Aim 1. Define the mechanism by which iron deficiency potentiates inflammation—In a mouse model of sterile inflammation we will examine the effect of ID on macrophage phenotype, the NF-kB pathway and systemic inflammation. Aim 2. Determine the mechanisms by which iron deficiency potentiates inflammatory lung injury—We will characterize adverse synergy between ID and acute inflammation (LPS) in the lung, and determine how ID modifies the role of CREB transcriptional factor in acute lung injury. Aim 3. Determine the mechanisms by which iron deficiency potentiates inflammatory kidney injury—We will characterize adverse synergy between ID and acute inflammation (LPS) in the kidney, and determine how ID modifies ER stress in acute kidney injury. These concepts and findings will have both fundamental and translational impact and may be relevant to many inflammatory disorders. They may guide future clinical studies to examine the adverse interaction in human conditions, and to ameliorate inflammatory injury through more effective management of iron deficiency.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ ABSTRACT Obstructive sleep apnea (OSA), a life-threatening condition, affects ~10% adults in the United States. The condition is accompanied by multiple symptoms (autonomic, breathing, mood, and cognition), which are linked to increased morbidity and mortality and decreased quality of life. The impaired functions likely result from brain changes in sites that mediate these regulations, but the pathological mechanisms contributing to brain changes are unclear. Intermittent hypoxia (IH), a primary characteristic of OSA, induces oxidative stress, leading to excessive production of reactive oxygen species (ROS) that encourages neuroinflammation and activates immune and glial cells; that sequence leads to regional increases in brain temperature, contributing to tissue damage. Moreover, IH induces mitochondrial dysfunction, altering metabolites, including the N-acetyl- aspartate (NAA; neuronal integrity), choline (Cho; membrane metabolism/integrity/turnover), creatine (Cr; energy metabolism), myo-inositol (MI; astrocyte proliferation/osmotic balance), and lactate (Lac; anaerobic metabolism). Also, antioxidants, including glutathione (GSH) that plays a significant role against oxidative stress, regulate neuronal/cellular protection from excessive ROS. However, the distribution of regional brain temperatures, metabolites, and antioxidant status in OSA is unknown, leaving a gap in knowledge of sources of injury that can be examined non-invasively with the 3D Echo Planar Spectroscopic Imaging (3D EPSI) and the MEshcher–GArwood Point RESolved Spectroscopy (MEGA-PRESS). Therefore, using 100 moderate-to- severe OSA and 100 age- and sex-matched controls, the specific aims are to: 1) examine regional brain temperatures, using 3D EPSI, in OSA and controls; 2) assess whole-brain metabolites (NAA, Cho, Cr, MI, and Lac), using 3D EPSI, antioxidant (GSH) levels from the posterior and anterior cingulate and anterior insula, using MEGA-PRESS, in OSA and controls; 3) determine the relationships between brain temperatures and Lac and MI levels with disease severity in OSA adults; and 4) assess regional brain temperature and metabolites changes, using 3D EPSI, after 6-months in OSA with and without continuous positive airway pressure (CPAP) compliance. In summary, the objective is to examine IH-induced oxidative stress and mitochondrial dysfunction processes contributing to brain changes in OSA, reflected as changes in regional brain temperatures and metabolites, and antioxidant in multiple brain sites, links between brain temperature, metabolites, and disease severity, and assess if the CPAP normalizes such changes in OSA. The findings have important implications for identifying interventions (nonsteroidal anti-inflammatory drugs, antioxidant, and Cr therapies as used in other conditions) to fully rescue brain changes in OSA with and without CPAP in those sites, which will benefit to dysfunctions, especially in non-CPAP compliant OSA or no treatment, and could dramatically improve the morbidity, mortality, and life quality.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract Colorectal cancer (CRC) is the second most common cause of cancer-related mortality in the United States (U.S.). Although several effective prevention and early detection screening methods are available, screening remains underutilized. The most feasible CRC screening modality for average-risk individuals in low resource settings, such as Federally Qualified Health Centers (FQHCs), is the stool-based fecal immunochemical test (FIT). One challenge of using FIT, however, is that follow-up colonoscopy is required after an abnormal (i.e., positive) result to check the colon and rectum for polyps or cancer. The benefit of FIT screening on CRC risk and mortality is realized only if timely colonoscopy is achieved. FQHCs are an integral component of the safety-net health system and represent a strong infrastructure for health service delivery research. Prior research suggests that only 18-57% of FQHC patients with abnormal stool-based screening results receive colonoscopy. There is limited research that focuses on identifying effective strategies to improve follow-up after an abnormal FIT result. In addition, much of the prior research has been conducted in integrated healthcare settings, although most care for underserved populations is delivered through a patchwork of public and private primary care providers, specialty providers and hospital settings. Therefore, it is paramount to understand how to increase follow-up of abnormal FIT results in these complex and real-world settings, which can yield findings with more relevance for the most vulnerable patient subgroups. In the proposed application, we will utilize a pragmatic, cluster randomized trial design within one of the largest FQHCs in the nation to test the effectiveness of a multilevel and multicomponent intervention (patient, provider, health system) that targets several barriers to follow-up in both the FQHC (primary care) and GI specialty care settings. The intervention is informed by prior research and is innovative in that it addresses barriers to follow-up colonoscopy in non-integrated primary care settings and within GI specialty care settings and strengthens care coordination between FQHCs and GI providers. The study will be conducted in 6 clinics within the same FQHC system. Three sites will be randomized to usual care and three to the intervention condition. The specific aims are to: 1) compare the effectiveness of the multilevel intervention to the usual care condition on receipt of a colonoscopy within 6 months of an abnormal FIT result (primary outcome); 2) systematically assess the quality of and challenges to intervention implementation to understand the feasibility and relative importance of intervention elements; and 3) measure the cost of intervention implementation to inform potential for dissemination and spread. The proposed study addresses important gaps in CRC prevention and control research by focusing on completion of the CRC screening process in a medically underserved population. Findings have the potential to change clinical practice and can be adapted for other FQHCs and clinical settings that face similar challenges in managing patients with abnormal FIT results.

NIH Research Projects · FY 2024 · 2024-09

Abstract Cushing Disease (CD) a life-threatening “orphan disease” with an annual US incidence of ~8 cases per million. It is caused by an adrenocorticotropic hormone (ACTH)-secreting pituitary adenoma, which drives excess adrenal-derived cortisol production. The annual health care cost of CD patients is > 7 times higher than average patients, and there is a large unmet medical need in treatment for this “orphan disease”. In a systematic high throughput screen of 200,000 compounds, we identified the dual PI3K + HDAC inhibitor, fimepinostat (CUDC-907) as an incredibly potent inhibitor of murine and human corticotroph tumor POMC transcription and ACTH secretion in both in vitro and in vivo models of CD. Supported by our pre-clinical data, we hypothesize that Fimepinostat suppresses pituitary corticotroph tumor growth and ACTH production to normalize cortisol levels in patients with CD. We now propose a pilot, short-term (4 weeks) phase II single- center study to demonstrate the safety and efficacy of Fimepinostat in the treatment of patients with de novo, persistent, and/or recurrent CD recruited at the University of California, Los Angeles. This Phase 2 open-label adaptive study has 2 specific aims: Aim 1 will assess the efficacy and Aim 2 the safety respectively of Fimepinostat in 20 subjects with de novo, persistent, or recurrent CD from UCLA outpatient clinics. Subjects will be randomized to either Fimepinostat 60mg (two 30mg capsules once a day, 10 subjects) or 30mg (single 30mg capsule daily in 10 subjects). Fimepinostat will be administered days 3-7 of each week and then 2 days off for a total of 4 weeks. Drug efficacy in both treatment arms will be determined based on the response rate of subjects exhibiting normalization and/or >50% reduction of 24-hour urinary free cortisol (24h UFC) levels (mean of 3 consecutive samples) after treating 10 subjects in each treatment dose arm. As further efficacy assessment, we will compare several secondary endpoints including normalization of 24h UFC (days 23-28 inclusive), plasma ACTH, serum and salivary cortisol levels (D29) to the subject’s baseline levels (Days -1 to - 5). Changes in clinical signs and symptoms of hypercortisolism, including reductions in body weight, body mass index, and blood pressure along with changes in health-related QOL and depression scores from baseline to 4 weeks will also be assessed although we acknowledge that some of these parameters may take longer to change than our 4 week study. A DSMB will carefully monitor all aspects of safety and assess any adverse and/or serious adverse events. This study will determine the most effective and least toxic dose regimen of Fimepinostat in the treatment of CD and pave the way for larger longer-term clinical trials in Cushing disease.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Despite advances in cancer treatment, the five-year survival rate for pancreatic ductal adenocarcinoma (PDA) is 12%. Obesity has been linked to PDA risk epidemiologically, implying that diet and metabolism have roles in PDA tumor initiation, but the biological basis for this correlation is not clear. Data from our labs shows that high fat diet (HFD) can accelerate PDA development in a Ptf1a-Cre/LSL-KrasG12D (KC) mouse model of PDA. Further, recent work has shown that acute inflammation can epigenetically prime acinar cells in the pancreas for tumorigenesis. We hypothesize that HFD, which has been clearly demonstrated to alter inflammation in a range of tissues, can similarly prime the pancreas to promote or prevent tumor development. The two aims in our proposal investigate effects of two HFDs chosen specifically because they are enriched for classes of fatty acids known to have distinct biological effects. Aim 1 examines the effects of a HFD comprised of lard (HFLD), which contains mostly saturated long chain fatty acids that are associated with obesity, on PDA tumorigenesis. To study how HFLD may prime the pancreas for tumor growth, we will use scRNA-seq and scATAC-seq to define dietary effects on healthy pancreas. We will also leverage our newly developed CYTO-Tag mouse to rapidly isolate acinar cells for metabolomics, thus allowing us to draw connections between changes in gene expression, chromatin structure, and metabolism in acinar cells in the context of HFLD. We will then use a tamoxifen- inducible Ptf1a-CreERT/LSL-KrasG12D (iKC) model to study the effects of HFLD on tumor growth, and use an orthotopic tumor model generated from HFLD-conditioned iKC acinar cells to distinguish between acinar cell- intrinsic and -extrinsic tumor growth mechanisms. In Aim 2, we will examine the effects of a HFD comprised of coconut oil (HFCD), isocalorically matched with the HFLD, on PDA tumor growth. Coconut oil is comprised of medium chain saturated fatty acids. It is a common dietary fat “swap” patients use when attempting to eat healthier, particularly in a ketogenic diet, but the effects of HFCD on PDA tumor growth are not known. We will use scRNA-seq, scATAC-seq, and our CYTO-Tag mouse to characterize the effects of HFCD on healthy pancreas, followed by tumor growth experiments in iKC mice. We will begin to pinpoint mechanisms underlying any observed effects on tumor growth through studies in orthotopic tumor models using HFCD-conditioned acinar cells. Aim 2 will define the effects of HFCD on tumor growth for the first time, and because our HFLD and HFCD are isocaloric, we will be able to directly compare effects of these diets on tumor growth. Understanding the molecular basis underlying differential effects of compositionally distinct HFD will enable more informed dietary recommendations for patients to potentially reduce cancer risk.

NSF Awards · FY 2024 · 2024-09

Consistency and reliability are necessary for algorithms used for data analysis and high-consequence decision-making. The project will contribute to this goal by developing algorithms for time-series analysis and anomaly detection that expand capabilities of understanding subtle features from multiple distinct data sources. In addition, this work will advance the spatial reasoning abilities of artificial intelligence algorithms which could be applied to other engineering or scientific problems. The project will train PhD students through involvement in the research. The aim is to develop mathematical algorithms for forecasting time-series, predicting spatial dynamics, and detecting anomalies using a multimodal transformer-based model. The project will construct methods for analyzing systems that switch dynamics or change behaviors. This can be applied to downstream tasks such as data analysis and anomaly detection. The research will address how to utilize contextual information with time series for more reliable predictions and how to consistently incorporate multiple pieces of information and observational modalities into prediction and anomaly detection. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Alphaviruses are mosquito-borne viruses that can cause arthritis and fatal encephalitis in humans with no approved therapeutics. Encephalitic alphaviruses, like Sindbis (SINV) and chikungunya (CHIKV) viruses, must cross the barriers protecting the central nervous system and therefore are neuroinvasive. The central nervous system is primarily protected by tightly connected brain microvascular endothelial cells (BMECs), pericytes, and astrocytes, which together constitute the blood-brain barrier (BBB). Genetic determinants of neuroinvasion reside in the alphavirus E2 glycoprotein and host factors that interact with the E2 glycoprotein have been identified. However, the relationship between neuroinvasive residues on E2 and the cell-specific expression of host interactors are not well characterized in the context of neuroinvasion. Our lab models the BBB by using pluripotent stem cell derived BMECs (iBMECs) that recapitulate the in vivo neuroinvasive phenotypes of alphaviruses and flaviviruses. Using this model, we have demonstrated that alphavirus neuroinvasion correlates with the ability to efficiently infect BMECs. Therefore, we hypothesize that the alphavirus E2 glycoprotein must interact with a BMEC-specific host factor to efficiently infect BMECs and cross the BBB. We will first determine how previously identified neuroinvasive E2 residues on SINV and CHIKV confer the ability to efficiently infect BMECs. Furthermore, we will identify the host factor that neuroinvasive alphaviruses hijack for efficient BMEC infection and characterize its interactions with E2. This work will elucidate virus-BBB interactions that facilitate neuropathogenesis, providing a foundation for therapeutic research. Under the fellowship training plan, I will advance my abilities in experimental design and science communication by executing, presenting, and publishing the results of this project, as well as through coursework and teaching as required by my doctoral program. My research training will take place in a rigorous and multidisciplinary scientific environment at the University of California, Los Angeles, where I have access to all the materials and expertise necessary for the completion of this project.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The overarching goal of the research presented in this application is to reveal the fundamental and conserved genetic and epigenetic programs that encode and transmit a memory of alcohol exposure for several generations. In mammals, in utero ethanol exposure is associated with an array of well-characterized neurobehavioral issues. However, there is mounting evidence in a variety of model organisms that some adverse neurological features are also detectable in the third generation following exposure, indicating a transgenerational effect. Alcohol also has a clear epigenetic impact and directly contributes to the modification of the epigenome. Nevertheless, it is unclear how the memory of ethanol exposure persists in the nervous system across generations. Here, I combine the tractability and conservation of the model system Caenorhabditis elegans with state-of-the-art epigenomic analyses, classical genetics, and behavioral approaches to shed light on the mechanisms of epigenetic memory of alcohol exposure and its transgenerational behavioral effects. My preliminary data shows that ethanol exposure (1) leads to increased histone acetylation in directly exposed animals; and that (2) ethanol seeking behavior is increased until at least the third generation, paralleling the increased alcohol consumption observed in rodent models and humans. Whether these observations are connected, and ethanol-mediated acetylation is directly responsible for the altered transgenerational impact on behavior is unknown. The specific altered acetylation marks and their distribution across the neuronal genome are also unknown. Thus, the driving hypothesis of this project is that neuronal histone hyperacetylation persists across generations to drive ethanol-induced transgenerational behavioral effects. In Aim 1, I will use a histone multiplex assay, immunofluorescence, and CUT&RUN sequencing of FACS-sorted neuronal nuclei to identify the neuronal epigenetic changes accompanying exposure to ethanol. In Aim 2, I will pharmacologically inhibit and perform neuron subtype-specific RNA interference of histone acetyltransferases and demethylases to determine the role of neuronal histone acetylation in ethanol’s transgenerational behavioral effects. This proposal has the potential to establish the impact of ethanol exposure on neuronal histone PTMs and their role in transgenerational behavioral effects. By applying genetic, epigenetic, and pharmacological approaches in a relevant and highly tractable organism, this proposed work will advance our knowledge of the epigenetic memory of alcohol exposure and its transgenerational behavioral effects. I will carry out these studies in Patrick Allard’s lab at UCLA, a respected expert in environmental epigenetics, with support from co-sponsors Xia Yang, an expert in epigenomics, and Kelly Huffman, an expert in fetal alcohol spectrum disorders. The environment at UCLA will also provide excellent intellectual, technical, and professional training. This will aid me in achieving my career goal of running an independent research lab studying the neuroepigenetics of chemical exposures.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY A major obstacle for the identification and characterization of new therapies for migraine and pain has been limitations of the animal models used to study these therapies. We have developed a novel approach that addresses this obstacle: a minimally invasive skull-attached microchip-based system and video recording/analysis system that enables continuous recording and triggering of neurovascular activity and behavior over months. We will further develop and validate this approach as a novel platform for the discovery and characterization of migraine therapies. We will: 1. Quantify a comprehensive set of neurovascular and behavioral responses to established human migraine triggers and cortical spreading depression (CSD) using the microchip/video platform. 2. Use the microchip/video platform to characterize the effects of acute migraine therapies on the response to established migraine triggers and CSD to validate the platform as a useful tool for the discovery and characterization of acute therapies 3. Use the microchip /video platform to characterize the effects of migraine preventive therapies. Key findings will be independently replicated at two different institutions. The platform described in this proposal represents a significant advance in our efforts to find new approaches to the treatment of migraine, a highly prevalent and disabling nervous system disorder.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Urethral defects requiring urethroplasty occur in children and adults secondary to congenital, traumatic, infectious, and malignant conditions. Current tissue sources for urethral replacement are limited by donor site morbidity and lack of optimal tissue characteristics to support lifelong voiding and penile erections. A subsequent high risk of short- and long-term urethroplasty complications highlights the need for an improved tissue alternative with a bioinspired design. The goal of this proposal is to engineer highly elastic, biomimetic, three dimensional (3D) bioprinted multi-layered urethral tissue constructs by combining novel bioinks, made of a human protein and decellularized matrix, with an innovative 3D bioprinting strategy. This research plan addresses key design requirements: 1) achieving target elasticity by layer in a suturable construct, 2) incorporating critical biological cues to enhance wound healing and vascularization, and 3) applying a 3D bioprinting technique to create optimized properties by layer with a recapitulation of the native urethral layered structure. Our overall hypothesis is that these novel 3D bioprinted constructs made from methacrylate human recombinant tropoelastin (MeTro), a photocrosslinkable human-based elastomeric hydrogel, and bladder decellularized matrix (BAM), that are designed to meet targeted mechanical and 3D structural parameters will improve suturability, early urinary tract function, and local tissue regeneration as compared to unseeded scaffold urethroplasties. In Aim 1, MeTro and BAM bioinks with mechanical and structural properties that mimic native urethral tissue will be engineered. Then, the designed bioinks will be 3D bioprinted to form cell-laden bi-layered patch constructs containing two primary lower urinary tract cell lines: urothelium and smooth muscle cells. In Aim 2, the in vivo efficacy of the engineered cell-laden MeTro/BAM bioprinted constructs, in seeded and unseeded configurations, will be applied to a rat patch urethroplasty model, investigating biologic and functional outcome parameters. Put together, this research strategy will engineer finely tuned elastic 3D printed biomimetic constructs with target mechanical and 3D structural parameters derived from urethral tissue analyses to maximize future clinical translatability.

NSF Awards · FY 2024 · 2024-09

The mechanical properties of cells play a crucial role in various biological processes, including cell adhesion, migration, and differentiation, and are essential for understanding disease progression. This project aims to develop an innovative OptoMagnetic Twisting Cytometry (OMTC) system to quantify the dynamic viscoelastic properties of 3D tissue samples. The OMTC system enables simultaneous measurements over a large field-of-view, advancing our understanding of the mechanical properties of live tissues. This groundbreaking technology has the potential to significantly impact disease diagnosis, treatment monitoring, and tissue engineering. By bridging the technological gap in current methodologies, the project aligns with NSF's mission to promote the progress of science and contribute to the national interest. The broader impacts of the project include advancing healthcare and biological research and disseminating knowledge through social media, conferences, publications, and courses. The OMTC system quantifies the local dynamic viscoelastic properties inside a 3D tissue sample by detecting the rotational movement of microparticles embedded within the tissue. This detection is achieved by capturing orientation-dependent light scattering and fluorescence signals from anisotropic microparticles. An optical scanning system that utilizes the focal plane scanning emission (FPSE) concept will be constructed to provide a collimated light beam with uniform intensity distribution for large volume directional illumination. This method eliminates the need for high numerical aperture optics for precision measurement and allows for simultaneous measurements of hundreds of thousands of microparticles within a large field-of-view using low-magnification objective lenses. The OMTC system can detect both the 3D positions and 3D orientations of microparticles. By analyzing the orientation changes before and after magnetic actuation, the local viscoelastic properties near each microparticle can be determined. Integrating data from microparticles randomly distributed within the tissue allows for mapping the dynamic viscoelastic properties of a 3D tissue sample. The developed OMTC system will be applied to study the dynamic processes of antitumor immunity by CAR-T cells in 3D tumor spheroid models. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-09

Acute ischemic stroke (AIS) is a leading cause of death and disability for which treatments are limited to reperfusion therapies, including intravenous thrombolytic (lytic) and endovascular thrombectomy (EVT). Many patients are not candidates for these therapies, and amongst those who receive them, the rate of excellent outcome remains low; only 20-30% are free from disability at 3 months post-stroke. Therefore, there is a critical need to develop additional novel therapies for patients with AIS. Cathodal transcranial direct current stimulation (C-tDCS) is a non-invasive inhibitory neuromodulatory technique that applies a weak electrical current via scalp electrodes. In animal models of acute cerebral ischemia, C-tDCS salvages penumbra (ischemic tissue at risk of infarction), both through direct neuroprotection by inhibiting peri-infarct excitotoxicity and through collateral perfusion enhancement by inducing vasodilation. C-tDCS has many advantages; it is a regionally directed therapy that instantly reaches maximum local concentration, and via high-definition (HD) electrode montages, electrical field’s spatial focality can be enhanced to target each patient’s ischemic tissue only. In our pilot study using HD C-tDCS, we showed that HD C-tDCS can be efficiently applied in AIS, and we observed promising signals of beneficial effects upon imaging biomarkers of neuroprotection and collateral enhancement, including penumbral salvage, improved perfusion, and cerebral blood volume enhancement. Therefore, we propose conducting a multi-site (3 sites), phase 2a, randomized, sham-controlled, dose-optimization study of HD C-tDCS as a neuroprotective and collateral enhancing treatment for AIS with and without lytic. The primary aim of the study is to identify, among six HD C-tDCS dose tiers, an optimal dose regimen that shows adequate safety and efficacy using imaging. Up to 120 AIS patients with cortical strokes and salvageable penumbra who are ineligible for EVT will be enrolled from UCLA, Johns Hopkins, and Duke. According to lytic eligibility, patients will be enrolled either in the lytic-receiving or non-lytic group and subsequently randomized 5:1 to active stimulation vs. sham. The study utilizes an adaptive Bayesian design with bivariate endpoints as its escalation-de-escalation rules. The primary imaging safety rule of radiographic intracranial hemorrhage probability ≤40% and the primary imaging biomarker rule of substantial penumbral salvage probability ≥70% will determine the pace and occurrence of escalation-de-escalation through the dose regimens. Secondary objectives include the effect of HD C-tDCS on additional imaging efficacy biomarkers of neuroprotection and collateral enhancement (hypoperfusion region volume, cerebral blood volume, and infarct growth), and other safety/tolerability endpoints. We will also explore the clinical efficacy of HD C-tDCS. The study primarily aims to find an optimal dose that meets an adequate threshold of safety and efficacy for future testing in larger randomized phase 2b/3 clinical trials. If proven safe and efficacious, HD C-tDCS has the potential to be used as a standalone treatment in patients ineligible for reperfusion therapies and as a synergistic treatment, improving outcomes of AIS patients.

NSF Awards · FY 2024 · 2024-09

By developing a framework to study higher order interactions, i.e., simultaneous interactions, the funded work will provide novel tools to analyze complex systems. The COVID-19 pandemic was challenging to control because people could catch the disease from accumulating many short exposures to multiple infected people, i.e., from higher order interactions, which are rarely considered in epidemiological models. Similarly, the efficient transfer of goods was another casualty of the pandemic due to supply-chain disruptions. Higher order interactions, in which goods are exchanged simultaneously, can substantially expedite the transfer of goods and increase the robustness and resilience of supply-chains to disruptions. The general framework that will be developed in this grant will use a tractable biological system to develop mathematical tools to study the causes and consequences of higher order interactions. The mathematical models and tools developed will be general, to allow application to other systems, such as communication, disease transmission, and social learning. Public health and bioeconomics are two examples of fields that can benefit from the funded work. The work will be published in general journals with a wide interdisciplinary readership and the analysis code will be made publicly available. Both PIs have a strong track record of recruiting and facilitating the success of students from groups that are unrepresented in the sciences and this commitment to mentoring a diverse population of trainees in interdisciplinary work will continue. To further disseminate the work to the general public, podcast episodes will be produced and distributed widely. Collective outcomes, such as the social behavior of animals, emerge from interactions among system components. While substantial work has been devoted to examining the intricate network of interactions among animals, these interactions are described and analyzed as dyadic events. However, multiple individuals can interact simultaneously. For example, an alarm call is broadcast to multiple individuals at once rather than through multiple one-on-one interactions. Despite the important conceptual and functional differences between dyadic and higher order interactions, there are only few methodological approaches that emphasize the higher order nature of social interactions. The proposed work will examine the causes and consequences of higher order interactions, and the feedback between them, by adapting and implementing existing mathematical tools from algebraic topology, simplicial sets, in novel ways. Specifically, the aims include to determine the conditions under which higher order interactions emerge; to examine the consequences of higher order interactions; and to investigate feedback between causes and consequences of higher order interactions to uncover potential evolutionary pathways for their emergence. Social insects are an especially powerful system for examining the questions in the proposal because of the profound fitness consequences of interactions among individuals for the group. Therefore, the proposed work will use foraging and food transmission of Argentine ants (Linepithema humile) as a model system to examine the internal and external causes and consequences of higher order interactions. Project outcomes will enable innovative approaches to fundamental and generalizable questions which are currently beyond our reach. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

In today's data-driven world, many complex systems can be represented as interconnected networks or graphs. This project aims to develop new methods for analyzing, generating, and optimizing these graph structures, with potential applications in areas such as social network analysis and molecular design. By improving the ability to learn from and work with graph-structured data, the project is expected to provide new tools for researchers across various scientific fields. The proposed research contributes to advancements in areas such as drug discovery, network analysis, and modeling of physical systems, offering new ways to approach complex problems in these domains. This project also offers research training opportunities for undergraduate and graduate students. The project focuses on four main research areas: (1) developing more expressive and efficient graph neural networks, (2) creating improved generative models for graphs, (3) applying graph learning techniques to optimization problems, and (4) exploring the use of graph neural networks for discovering physical relations. The interconnected research thrusts aim to improve the capabilities of machine learning models based on graphs, laying the groundwork for solving complex graph-related challenges. The project will produce new mathematical and statistical tools, theoretical frameworks, and assessment methods for learning from graphs. The work is expected to advance graph learning techniques and their applications in scientific fields, providing researchers with new ways to handle data structured as graphs. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

Robots capable of navigating unstructured terrains in diverse environments, such as water and land, are crucial for many real-world applications. While soft robots can navigate challenging environments like narrow tunnels and rough surfaces due to their flexibility, most current designs are limited by slow speeds, reliance on ties to the base unit (i.e., tethered), and use in only one type of environment, such as land or water. Additionally, soft robots are time-consuming and expensive to create compared to rigid robots, which benefit from centuries of innovative generation. This project aims to create a new class of untethered, reconfigurable (i.e., able to change shape), and multimodal amphibious soft robots (URSoRo) assisted by a machine learning (ML) design tool to overcome these limitations. These robots will leverage a new class of soft electromagnetic (EM) actuators that can operate in more than one state, enabling them to swiftly adapt to challenging environments. This project will leverage the reconfigurability of soft robots for environmental adaptation and promote their practical applications, such as search and rescue operations, monitoring of animals and plants, and inspection of infrastructures in extreme environments. Additionally, the project will contribute to an annual inter-university soft robot competition across the United States and integrate findings into graduate-level courses on soft robotics at the University of Michigan, Ann Arbor, and the University of California, Los Angeles. This project addresses two primary challenges in soft robotics: designing shapes and achieving bistability in soft actuators while maintaining a simple, low-cost fabrication process, and tightly integrating and engineering untethered reconfigurable soft robots with fast multimodal locomotion. The research will develop a soft bistable EM actuator with high force output (∼0.4N), high activation frequency (>30 Hz), and the capability to be powered by miniaturized onboard electronics (<15 g). An ML-assisted physics-based simulation tool will be developed to guide the design, fabrication and robotic integration of these EM bistable actuators, enabling a fully planar rapid fabrication process. Liquid metal embedded elastomers will be used to enhance both thermal management and electromagnetic field generation, boosting the actuator's performance. Overall, this project will result in a new class of untethered soft robots driven by soft bistable EM actuators, alongside ML-assisted physics-based modeling and design tools, achieving an unprecedented combination of speed, size, mass, and reconfigurability. By addressing these technical challenges, it will contribute to the field of robotics with versatile, efficient, and cost-effective solutions for creating soft robots with rapid reconfiguration and advanced locomotion performance in unstructured and diverse real-world environments. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Interstitial lung diseases (ILD) are a large group of lung diseases that are characterized by chronic, progressive pulmonary fibrosis. We hypothesize that in all ILDs the progressive fibrosis is driven by ongoing injury/stress in the airway epithelial cells that set up crosstalk with their neighboring mesenchymal cells resulting in a chronic wound healing process that alters the matrix and changes the cellular niche. The goal of the proposed research is to address how interactions between the epithelial, mesenchymal and matrix compartments drive progression of fibrosis in all forms of ILD. We are proposing to study these interactions between the epithelial, mesenchymal and matrix compartments, which is now feasible with spatial transcriptomics, single cell secretion protein analysis, and small region proteomics. Most studies have looked at end stage disease and fibrotic lesions where most of the epithelial cells are lost and consequently the initiating and propagating factors are not well understood. We propose to study Interstitial Lung Abnormality (ILA) lesions, which represent very early ILD lesions, early stage ILD lesions when patients start to become symptomatic, and late stage ILD regions that still contain epithelial cells so we can study their interactions with mesenchymal cells and matrix. To test our hypothesis, we have developed a 3D scaffolded lung cell co-culture model system with progressive fibrosis that closely models these cellular compartments in ILD tissue. We propose the following aims: Aim 1: To understand epithelial-mesenchymal interactions in the airway that drive fibrosis using spatial transcriptomics. We will use our biobank of ILD and ILA samples and prospectively collect ILD and ILA patient samples to profile the airway epithelium and underlying fibroblasts from bronchioles to distal airspaces with spatial small region transcriptomics (GeoMx) and single cell spatial genome-wide transcriptomics (Stereoseq). Aim 2: To understand epithelial, mesenchymal and matrix interactions in the airway that drive fibrosis. Aim 2a: We will use the secretory-single cell profiling (Sec-seq) technology to capture single airway epithelial cells, from fresh ILA and ILD tissue and trap the secretions from each cell to identify paracrine factors. Aim 2b: We will use nanoscale small region proteomics to identify proteins in the adjacent epithelial, mesenchymal and basement membrane matrix compartments from ILA and ILD samples. Aim 3: To identify mechanisms by which persistent airway injury induces progressive fibrosis by using cell co- culture models of iPSC derived AT2 cells with specific mutations associated with familial IPF, in conjunction with primary healthy or IPF lung fibroblasts. These powerful reductionist models allow the study of specific epithelial- mesenchymal-matrix interactions that drive progressive fibrosis. We have assembled a team with expertise in spatial transcriptomics and ILDs (Gomperts), novel bioinformatics pipelines and integration of genomics data (Plath), regional small-scale proteomics in ILDs (Clair), iPSC-derived AT2 disease modeling and ILDs (Kotton) and bioengineered models of ILD (Gomperts).

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Facial paralysis from stroke or other neurological disorders often causes loss of the eye-blink reflex, leading to pain and visual disability. Eyelid dysfunction leaves the eye chronically exposed, which is not only painful, but is fundamentally incompatible with functional vision. Further, because eyelid movement plays a critical role in facial expression and human communication, loss of natural eyelid motion can also have negative social and cultural implications. Unfortunately, current surgical management strategies have major limitations in both functionality and appearance. In theory, dynamic natural blink restoration via a facial neuroprosthesis would be an ideal solution; unfortunately, this ideal has proven elusive, due in large part to a lack of knowledge regarding the neurophysiological mechanisms that enable eyelid function. There is a critical need for neuroprostheses that reproduce functionally complete and aesthetically natural eye closure, blink, and other behaviors. In response to this need, our long-term goal is to advance a novel class of neuroprostheses that are informed by a deep understanding of the fundamental neuromechanics of the muscle that controls the eyelid. To achieve our long- term goal, we will first carry out fundamental neuroscientific studies to establish the currently-unknown mechanisms that link segmental muscle activation to eyelid motion and function. The innovation of this work lies in our ability to measure intramuscular activation and three-dimensional eyelid kinematics with unprecedented precision and resolution. This sets our work apart from all other prior research into eyelid function, and will allow us to develop the first predictive dynamic neuromuscular model of the eyelid. In the present work, our objective is to study the neurophysiology of how activation sequences and intensities produce blink and other eyelid behaviors under both healthy and pathological activation. We will accomplish this by first studying eyelid function in persons without paralysis during a range of eyelid behaviors, including spontaneous blink, reflexive blink, and forced closure. As the participants perform these behaviors, we will record high-resolution intramuscular EMG from multiple points within and around the eyelid, while simultaneously tracking the three-dimensional motion of several points along the eyelid margin in high definition. We will use these data to implement a mechanistic neuromuscular model of the eyelid musculature, which can then inform where and when stimulation from future neuroprostheses should be delivered. We will then repeat these experiments in a group of persons with partial facial paralysis, to study the mechanisms by which eyelid function can be compromised. Upon completion of this work, we expect to have established the mechanistic basis for model-informed facial neuroprostheses that restore natural blink. These results are expected to provide the foundation for development and evaluation of neuroprostheses with the potential to improve eye health, vision, and confidence for patients with facial paralysis.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Duchenne muscular dystrophy (DMD) is a lethal pediatric neuromuscular disorder that affects 1/5000 boys globally. This rare disease is caused by mutations in the gene encoding the dystrophin protein, often by disrupting the reading frame. Antisense oligonucleotide (ASO) therapeutics target the dystrophin pre- mRNA, induce exon skipping to restore the reading frame, and partially rescue dystrophin expression. Four ASO drugs have been approved by the U.S. Food and Drug Administration. However, their clinical efficacies are dismally low due to significant barriers in activity and delivery. To address these problems, we propose two innovative strategies for developing highly effective ASOs targeting dystrophin exons 44 and 45, which together could treat about 14% of DMD patients. Both strategies take advantage of the fact that RNA tends to fold into structures. In the first strategy, we design ASOs with tertiary interactions that dramatically expand the ASO-exon molecular interface, in addition to conventional Watson-Crick base pairing. The tertiary contacts enable the ASO to recognize both the sequence and the structure of the target exon, potentially driving higher affinity, specificity, and exon skipping activity. For exon 44 (Aim 1) we design the ASO sequence and backbone chemistry to generate tertiary contacts with a short hairpin in the pre-mRNA, which enhances binding affinity. The crystal structure of the ASO-exon complex reveals opportunities for creating additional interactions via chemically modified bases. In subsequent rounds of design and testing, we will synthesize modified oligos, characterize them structurally and biochemically, and measure exon-skipping activity in DMD patient-derived muscle cell culture. In the second strategy, we develop a bifunctional cell-penetrating peptide (CPP) that recognizes a unique hairpin adjacent to the binding site of existing ASO drug Casimersen in exon 45 (Aim 2). Conjugation of the CPP to Casimersen should enhance both delivery and target RNA recognition. In preliminary studies, we solved a high- resolution crystal structure of the exon 45 hairpin and discovered non-canonical base pairs that create unusual structural features in the major groove. We will computationally design CPP sequences to recognize the novel major groove conformation. We will co-crystallize candidate CPPs with the exon 45 hairpin and quantify their affinity and specificity. For promising leads, we will produce CPP-ASO conjugates and measure cell penetrance and exon 45 skipping activity in patient cell lines. We anticipate testing the best ASOs developed in this study in animal models in the future, with potential for clinical trials. The structure-based strategies can drive development of more effective ASO drugs for skipping other dystrophin exons, leading to increased access to precision therapies for this debilitating childhood disease.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract Anxiety disorders represent a significant mental health disease burden in the United States today, with roughly 1 in 3 Americans expected to be diagnosed with an anxiety disorder in their lifetimes. Current treatment strategies include psychotherapy and psychopharmacology, which are better than nothing, but still are insufficient. Novel brain stimulation techniques have emerged as putative alternatives, but these have drawbacks, namely imprecision and lack of ability to stimulate deep brain structures. Transcranial focused ultrasound (tFUS) suppression of the amygdala has the potential as an ideal therapeutic due to the combination of depth and precision. Today, unfortunately, it lacks validation of the focality of stimulation. Magnetic resonance acoustic radiation force impulse (MR-ARFI) imaging is a technique that can accurately image the tFUS focus by visualizing the micrometer displacement of tissue due to ultrasound. However, MR-ARFI has never before been shown in-vivo in the human brain. This proposal will show for the first time MR-ARFI imaging in humans. Therefore, in Specific Aim 1, MR-ARFI imaging will be acquired in an anthropomorphic phantom. These MR sequence parameters and ultrasound parameters will form the basis of Specific Aim 2, which will show in-vivo amygdala targeting that the location of the ultrasonic focus as predicted by MR-ARFI imaging will be in the area predicted by modeling software. In Specific Aim 3A and 3B, we will show that the more accurately the ultrasound targets the amygdala, the greater the reduction in amygdalar perfusion and anxiety rating scores will be. The described research will form part of the fellowship training plan, providing the fellow with training in MR sequence design, ultrasound parameter design, phantom MR imaging, human subjects imaging, and analysis of human imaging and behavioral data.. The entirety of the fellowship training plan (including the proposed research project) will take place at UCLA. It will supplement the fellow’s training as part of the UCLA-Caltech MSTP. The training plan will be jointly supervised by Drs. Susan Bookheimer and Martin Monti, forming the basis of the fellow’s dissertation. It will give him the skills and training to be an outstanding clinician-scientist.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Disorders of the peripheral nervous system, particularly those that affect motor function and pain sensitivity are major clinical challenges. Current medications are not curative and only address secondary symptoms of disease. To restore peripheral nerve function after injury, Schwann cells adapt to counteract pathological insults by undergoing a dramatic transformation to generate repair cells. This complex process, termed transdifferentiation, requires coordination between multiple signaling and gene regulation programs. During transdifferentiation Schwann cell myelination, metabolism, adhesion, phagocytic properties and the ability to activate the immune system are adapted to preserve neuronal survival and function. Transcriptional studies of nerve injury models have revealed that several canonical signaling pathways are altered after injury, including: NF-kB, Ras, TGFβ, WNT, and MAPK. Despite the clear upregulation of WNT signaling components, how WNT signaling contributes to the generation of the repair cell is unclear. Poor understanding of the molecular systems controlling repair cell formation and function is a critical barrier towards identifying therapeutic targets to treat nerve injury. We now have the genetic, sequencing and viral tools to dissect how signaling pathways individually contribute to each of the hallmark repair cell phenotypes. We hypothesize that WNT signaling mediates repair cell formation by promoting transdifferentiation and the epithelial-to-mesenchymal transition of repair cells. This grant will 1) provide functional tests of the role of WNT receptors and ligands in the control of repair cell formation 2) determine whether reduction of a WNT antagonist can restore peripheral nerve regeneration after injury. To accomplish these goals, I will leverage my training in disease models of peripheral neuropathy to gain experience in four critical areas: (1) viral vector design of CRISPR/Cas9 tools, and (2) RNA detection and quantification techniques, (3) mechanisms of cellular reprogramming, and (4) professional development. To achieve these training goals, I have assembled an exceptional group of mentors and scientific advisors with specialties in understanding the roles that glia play during development, injury and neurodegeneration, virus-mediated CRISPR/Cas9 delivery, and transcriptomics of myelinated tissues. With expert guidance from this group, the proposed experiments will address the cellular mechanisms of repair cell induction. The results from this application will serve as the basis for an R01 application to dissect how WNT signaling components orchestrate the changes in transcription required to reprogram mature Schwann cells into repair cells and test whether WNT intersects with other relevant intracellular pathways (e.g. MAPK, ERK) during Schwann cell reprogramming. .

NSF Awards · FY 2024 · 2024-09

In collisionless space plasma, mixing and exchanging different energies, e.g., magnetic field energy and charged particle kinetic energy, are controlled by the formation and destruction of plasma structures like current sheets. Current sheets are universal quasi-1D self-consistent plasma and magnetic field configurations that naturally form as boundaries between different plasmas or as current layers embedded within stretched magnetic field lines (typical configuration for the planetary magnetospheres). This project will systematically examine current sheet characteristics in Martian and terrestrial magnetospheres. These two systems differ significantly in plasma content and energies. The team will reveal details of current sheet formation and destruction associated with charged particle acceleration in different parametric regimes. This collaborative project between UTD and UCLA involves significant contributions from two PIs and two graduate students. The configuration and stability of an essential kinetic plasma structure, the current sheet (CS), determine the efficiency of magnetic energy storage, release, and transport in surrounding plasmas. These properties depend on plasma parameters (the ratio of plasma to magnetic field pressures, the ratio of bulk velocities to magnetosonic velocities, etc.). The main scientific goal of this project is to systematically characterize current sheet configurations in the Martian magnetotail (using MAVEN observations), compare these configurations with statistical results of the Earth’s magnetotail current sheets, and reveal the role of plasma parameters, as well as heavy ion contributions, in various configurations. To mitigate the uncertainties due to single-spacecraft measurements from MAVEN, this analysis will be supplemented by comparisons with kinetic simulations of the Martian magnetotail. To compare Martial and Earth’s magnetotail current sheets, we will use current sheet datasets from THEMIS and ARTEMIS, supplemented by Cluster and MMS datasets. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

The broader impact of this I-Corps project is the development of a biotechnology tool to increase gene editing efficiency and accelerate the development of cell therapies. Gene editing technology, a method for making specific changes to the DNA of a cell, is used to turn human cells into therapeutic cells. The method may be used as a potential treatment and cure for many diseases including cancer. Currently, however, this application is limited by the low efficiency of gene editing, which results in only a few percent of the cells being engineered successfully and becoming therapeutic cells. This low yield makes cell therapy one of the most expensive treatments and creates significant unmet patient demand. With increased gene editing efficiency, more therapeutic cells may be created, which may lower the manufacturing cost of therapeutic cells and help cell therapies treat more patients. This I-Corps project utilizes experiential learning coupled with a first-hand investigation of the industry ecosystem to assess the translation potential of a microfluidic device to increase gene editing efficiency. To achieve successful gene editing, materials used for gene editing need to be delivered into cells and have access to the target genes. However, many genes are densely packed and hidden within the chromatin, which makes them difficult to reach and leads to low editing efficiency. This technology tackles this issue through a mechanism called cell massage. Cells are gently squeezed through microchannels within the device, and the mechanical stimulation on the cell nucleus opens the chromatin structure temporarily. This opening allows the genes to be more accessible to gene editing materials. This solution has been shown to lead to a 10-fold increase in gene editing efficiency. The increased efficiency may lower the epigenetic barrier and make it easier for gene editing tools to reach and edit target genes. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.