Browsing by Author "Moreno, Julie, advisor"

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Open Access
Glial inflammation as a key regulator and therapeutic target for prion disease
(Colorado State University. Libraries, 2023) Hay, Arielle, author; Zabel, Mark, advisor; Moreno, Julie, advisor; Tjalkens, Ronald, committee member; Chanda, Soham, committee member; Santangelo, Kelly, committee member
Prion diseases are lethal neurodegenerative diseases characterized by the misfolding of the cellular prion protein, PrPC, into the infectious PrPSc. PrPSc accumulation in the brain contributes to the activation of microglia and the subsequent increase in reactive astrocytes, which together contribute to neuroinflammation. PrPSc aggregation triggers and leads to the dysregulation of a variety of cellular stress pathways, including the oxidative stress response, unfolded protein response, ubiquitin-proteosome system, autophagy and lysosomal degradation. Most critically, PrPSc contributes to neuronal toxicity and death, but the mechanism behind this is poorly understood. Prion diseases affect humans and a variety of mammalian species, with no available treatments. The majority of therapeutics developed to combat these diseases have targeted the prion protein itself. As these have been unsuccessful, it is time to turn our attention to treatments that target the cellular pathways and neuroinflammation caused by PrPSc accumulation in the brain. The overarching goal of this work is to identify glial-induced inflammation as a candidate for therapeutic intervention of prion diseases. We assessed the use of mesenchymal stromal cells (MSCs), which are potent regulators of inflammatory signaling and glial polarization, in cell culture and animal models of prion disease. Additionally, we investigate the role of a key inflammatory signaling pathway, Nuclear Factor-Kappa B (NF-κB) in microglial response to prion infection. Our findings both characterize contributions of specific glial cells to prion-induced inflammation, as well as uncovering novel targets for the treatment of prion diseases. First, we assessed the therapeutic potential of adipose-derived mesenchymal stromal cells (AdMSCs) in a cell culture model of glial prion infection. MSCs are known for their ability to migrate to sites of inflammation and produce immunomodulators. We evaluated the ability of cultured AdMSCs to respond to molecular factors present in brain homogenates from prion-infected animals. We found that these cells upregulate anti-inflammatory genes in response to both specific inflammatory cytokines and crude prion brain homogenates. Moreover, AdMSCs migrate towards prion brain homogenates in an in vitro model. Co-culturing AdMSCs with prion-treated BV2 cells or infected primary mixed glial cultures resulted in a significant decrease in markers of inflammation and disease-associated microglia and reactive astrocyte markers. These findings were independent of PrPSc, as AdMSCs had no effect on prion accumulation in mixed glial cultures. Collectively, these findings highlight AdMSCs as an intriguing candidate for modulating glial-induced inflammation in prion disease. Next, we evaluated AdMSCs in a mouse model of prion disease. Prior to delivery into prion-infected mice, AdMSCs were stimulated with TNFα, which we show increases their upregulation of anti-inflammatory molecules and growth factors. Stimulated AdMSCs were delivered intranasally to prion-infected mice every two weeks beginning from early in infection (10 weeks post-infection (wpi)) and ending late in infection (20 wpi). A cohort of mice was euthanized at various stages in infection, at 14 wpi, 16 wpi and 18 wpi. We show that AdMSCs are able to migrate throughout the brain when delivered intranasally, with the most cells being found in the hippocampus and thalamus. Although AdMSCs were not successful in improving behavior or increasing survival in prion-infected mice, they did induce changes in prion pathology at early time points in disease. A decrease was seen in inflammatory cytokines and markers of glial activation. No changes were seen in PrPSc accumulation or neuronal loss compared to untreated controls. However, at both 16- and 18 wpi, we identified significant changes in both glial numbers as well as morphology, indicating that AdMSCs attenuate reactivity in microglia and astrocytes. Together, these findings highlight AdMSCs as potent regulators of prion-induced glial inflammation, and warrants further investigation to optimize these cells as a treatment for prion disease. In addition to assessing therapeutics that decrease inflammation and reprogram glial cells to a homeostatic phenotype, we wanted to better characterize specific inflammatory pathways and understand how these were being regulated in glial cells in response to prion infection. NF-κB-related genes have long been identified in the brains of animal models with prion disease, but studies that have investigated its role in prion pathogenesis have focused on neurons and astrocytes. Microglia are critical innate immune regulators in the brain, and interact closely with both neurons and astrocytes to regulate inflammation and cell survival. Therefore, we saw an immediate need to characterize NF-κB signaling in microglia, and its contribution to prion-induced neuroinflammation. IKβ kinase (IKK) is a complex that responds to cell stressors and is critical for NF-κB signaling to occur. We utilized a primary mixed glial model containing wild-type (WT) astrocytes and IKK KO microglia. Upon infecting these mixed glial cultures with prions, we saw a drastic decrease in NF-κB-related genes compared to cultures containing WT astrocytes and WT microglia. Despite this, cultures containing IKK KO microglia still contribute neurotoxic signals that induce neuronal cell death. Moreover, we found that cultures with IKK KO microglia showed significantly more PrPSc accumulation, suggesting that these cells may have impaired autophagy. This work implicates microglial NF-κB-signaling and IKK as a potent inducer of inflammation and regulator of autophagy in prion disease.
Open Access
Metronidazole neurotoxicity
(Colorado State University. Libraries, 2021) Vick, Zaria Denise, author; Moreno, Julie, advisor; Legare, Marie, advisor; Bouma, Jerry, committee member; Tjalkens, Ronald, committee member
Metronidazole is a broad-spectrum antibiotic approved for clinical therapeutic use in veterinary and human medicine. Although the literature has reported neurotoxic unintended side effects with the use of this drug, these incidences occur in less than 1% of human cases making this instance rare. The mechanism of this neurotoxicity has not been fully elucidated, nor the susceptible population identified. We explore in this work that these susceptible populations are humans and animals with concurrent localized and/or systemic inflammation. Some proposed mechanisms are axonal swelling with increased water content due to toxic injury, vascular spasm with mild reversible localized ischemia, modulation of the gamma-aminobutyric acid (GABA) receptors within cerebellar and vestibular systems, RNA binding with inhibition of protein synthesis, and axonal degeneration. While these mechanisms offer some insights into the neurotoxicity, we propose a novel connection between cholesterol inhibition and the reductive activation of metronidazole resulting in poor glial myelination that explicates low dose neurotoxic clinical outcomes in vulnerable humans and animals with the use of this drug. In order to investigate this, we have implemented physiologically based pharmacokinetic computational models of a human, equine, murine, and rabbit with metronidazole exposure. Furthermore, in combination with computational techniques, we assess cellular and molecular analyses to address this neurotoxicity in a primary murine glial cell model. Additionally, we use liquid chromatography and mass spectrometry work in order to address the reductive activation of metronidazole. We then ask if inflamed glia are more susceptible to metronidazole-dependent neurotoxic outcomes. With these data, we offer insight into this elusive mechanism and will aid human and veterinary literature in a way that improves the quality of life of affected patients and better predicts populations vulnerable to this neurotoxicity.
Open Access
Skn-1, Nrf homolog, mediates cannabidiol cellular stress responsive effects in Caenorhabditis elegans
(Colorado State University. Libraries, 2023) Alsulami, Abdullatif M., author; Moreno, Julie, advisor; McGrath, Stephanie, committee member; LaRocca, Tom, committee member; Arnold, Olivia, committee member
Alzheimer's disease (AD) is a neurodegenerative disease that is affecting an increasing number of the aged population worldwide. AD is characterized by the accumulation of amyloid beta (Aβ) and tau hyperphosphorylation along with a failure in redox homeostasis. The hallmarks of neurodegenerative diseases include the increased generation of reactive oxygen species (ROS) which is tightly controlled by an antioxidant defense mechanism under physiological conditions. This research aimed to utilize various strains of the model organism C. elegans to understand the mechanism of cannabidiol at the cellular level in stressed models. The SKN-1 gene, the Nrf homolog in C. elegans, encodes for three different isoforms, skn-1a, skn-1b, and c. Skn-1b/c, which plays a role in oxidative stress, is negatively regulated by the repressor WDR-23. In C. elegans, skn-1a plays a role in proteotoxic stress through upregulation proteosome subunits and is negatively regulated by the abundance of proteosome complex protein. Results show that 10μM of CBD was able to activate isoforms of skn-1, skn-1a and skn-1b/c. The ROS scavenging activity of CBD was dependent on the presence of skn-1b/c. Furthermore, CBD's protective effects under proteotoxic stress were diminished in the absence of skn-1a. Further investigation will be conducted to identify the role of skn-1 in CBD's reduction of Aβ plaques.
Embargo
The neuroinflammatory nexus: glial dysfunction in the pathogenesis and therapeutic targeting of neurodegenerative and neurodevelopmental disorders
(Colorado State University. Libraries, 2025) Risen, Sydney J., author; Moreno, Julie, advisor; Tjalkens, Ronald, committee member; LaRocca, Tom, committee member; Nordgren, Tara, committee member
Chronic neuroinflammation is increasingly recognized as a fundamental driver of both neurodegenerative and neurodevelopmental disorders, linking immune dysregulation, glial dysfunction, and disease progression. In neurodegenerative protein misfolding disorders (NPMDs), including Alzheimer's disease, Parkinson's disease, and prion diseases, sustained microglial and astrocytic activation exacerbates protein aggregation, synaptic dysfunction, and neuronal loss, accelerating cognitive decline. Similarly, in neurodevelopment, aberrant inflammatory signaling during critical windows of brain maturation impairs synaptic formation, alters neurotransmitter systems, and predisposes individuals to long-term cognitive and behavioral deficits. Despite distinct manifestations, both disease categories share a pathological feature: a maladaptive neuroimmune response disrupting neural homeostasis. While neuroinflammation is widely implicated in these disorders, defining its molecular mechanisms, identifying therapeutic targets, and understanding environmental contributions remain critical research needs. This dissertation begins to address these gaps by investigating neuroinflammation as both a therapeutic target in NPMDs and a mechanistic link between environmental exposures and neurodevelopmental disruption. The first investigation evaluates SB_NI_112, a novel brain-penetrant RNA-based therapeutic designed to selectively inhibit NF-κB and NLRP3 inflammasome pathways, key regulators of glial activation and chronic neuroinflammation. Pharmacokinetic and biodistribution studies in small- and large-animal models were first conducted to assess SB_NI_112's CNS penetration and safety. These studies confirmed robust brain bioavailability (~30%) and a favorable safety profile, supporting its viability for therapeutic application. Following these findings, SB_NI_112 was tested in a murine prion disease model, where treatment significantly reduced microglial and astrocytic activation in disease-relevant brain regions, preserved hippocampal neurons, and mitigated neurodegeneration. These neuroprotective effects corresponded with improved cognitive performance in novel object recognition tasks, indicating functional preservation despite ongoing prion pathology. Notably, SB_NI_112 treatment extended survival, reinforcing inflammasome inhibition as a viable therapeutic strategy for NPMDs. These findings provide strong proof-of-concept for targeting neuroinflammatory pathways to slow disease progression and preserve cognitive function in neurodegenerative protein misfolding disorders. The second investigation examines the role of environmental neurotoxicants in triggering neuroinflammation and impairing neurodevelopment. Using a juvenile mouse model, this study demonstrates that chronic low-dose manganese (Mn) exposure (50 mg/kg via drinking water) first alters gut microbiome composition, depleting the relative abundance of beneficial Lactobacillaceae, and increasing the relative abundance of pro-inflammatory Erysipelotrichaceae, contributing to gut-brain axis dysfunction. These microbial shifts coincide with significant gliosis in the enteric nervous system, suggesting early neuroimmune activation at the gut interface. This inflammatory response extends to the brain, where widespread microglial and astrocytic activation is observed alongside disruptions in neurotransmitter production and metabolism, including altered dopamine and serotonin homeostasis. Functionally, these neuroimmune and neurochemical disruptions correspond with changes in behavior, indicating impaired neural processing. The presence of inflammatory lesions in the intestinal lining further implicates gut inflammation as a mediator of Mn-induced neurodevelopmental deficits. These findings highlight the systemic impact of Mn exposure, reinforcing the link between environmental toxins, neuroinflammation, and behavioral dysregulation. Together, these studies further support the growing body of evidence that neuroinflammation is a primary driver of neurological disease rather than a secondary consequence, reinforcing the need for targeted neuroimmune interventions. By examining shared inflammatory features across NPMDs and environmentally induced neurodevelopmental disruptions, this work provides additional insight into how glial dysfunction contributes to neurological pathology. These findings support the continued development of neuroimmune-modulating therapeutics, emphasize early intervention, and highlight the importance of environmental risk mitigation. By bridging molecular, pharmacological, and environmental perspectives, this dissertation contributes to the broader understanding of neuroinflammation in disease progression, challenging traditional distinctions between neurological disorders and providing a foundation for future studies. The implications extend beyond basic science, offering translational potential for clinical intervention, public health strategies, and regulatory policies to reduce the burden of neuroinflammatory disease.