Glioblastoma (GBM) remains highly resistant to immunotherapy due to a highly immunosuppressive tumor microenvironment (TME), underscoring the need for clinically relevant models to evaluate new therapeutic strategies. The mosaic analysis by dual recombinase-mediated cassette exchange (MADR) glioma model closely recapitulates the diffuse and infiltrative nature of human GBM. This model enables direct comparison of tumor-targeted and immune-directed therapies to determine how they reshape the tumor microenvironment. MADR glioma cells were intracranially implanted into C57BL/6 mice to generate tumor-bearing animals. Two therapeutic approaches were evaluated: EGFR inhibition (using doxycycline and ERAS-801), aimed at modulating tumor signaling and reducing immunosuppressive myeloid populations, and an LNP-formulated mRNA vaccine encoding gp100, designed to enhance antigen-specific immune activation and promote T-cell expansion within the TME. Preliminary findings indicate increased infiltration of CD4, CD8, and Thy1.1 T-cells in mRNA-treated mice, supporting enhanced immune activation. These observations provide a basis for analyzing immune expansion, immune cell co-localization, and spatial organization within the TME. Together, this work establishes the MADR model as a translationally relevant system for evaluating immunotherapies and supports combining EGFR inhibition with mRNA vaccination to improve therapeutic efficacy in GBM models for future translation studies.

Abnormal myocardial metabolism characterizes and may promote heart failure (HF), but its link to cardiac remodeling remains unclear. Our lab previously found that 2-hydroxyglutarate (2-HG) accumulates in multiple HF models. To test whether enhancing 2-HG clearance reduces remodeling, we over-expressed D-2-hydroxyglutarate dehydrogenase (D2HGDH) and L-2-hydroxyglutarate dehydrogenase (L2HGDH) to degrade both 2-HG isomers in mouse hearts subjected to transverse aortic constriction (TAC). Compared to mCherry and D2HGDH groups, L2HGDH over-expression significantly attenuated cardiac hypertrophy post-TAC. To investigate mechanisms, we generated L2HGDH and D2HGDH knockout (KO) mice and performed proteomics on L2KO and D2KO hearts. Cardiac proteins were analyzed by data-independent acquisition mass spectrometry (DIA-MS) using a DIA-NN pipeline. Proteomics identified 56 differentially expressed proteins in D2KO hearts (33 up-regulated, 23 down-regulated), while only 2 proteins were significantly altered in L2KO hearts, both down-regulated. Combined with metabolomics, pathway enrichment analysis suggests D2KO more severely depletes electron transport chain components, whereas L2KO has stronger epigenetic effects. Elevated 2-HG is a marker of cardiac remodeling; thus, this study examines its stereospecific roles and therapeutic potential.

Vocal fold (VF) scarring causes dysphonia and reduced quality of life but remains difficult to accurately diagnose with current noninvasive modalities. Dynamic Optical Contrast Imaging (DOCI) is a novel modality that measures tissue autofluorescence lifetime and wavelength-dependent properties and offers a biochemical approach to tissue differentiation. The DOCI system's ability to distinguish injured and uninjured VF tissue was tested by measuring autofluorescence lifetimes in a rabbit model (n=10) with unilateral VF injury and contralateral internal controls. Eight weeks post-injury, hemilarynges were excised for imaging and histological processing. Regions of Interest from injured and uninjured VFs were analyzed to extract normalized median lifetime values. Pentachrome-stained hemilarynges showed clear structural differences, with injured VFs displaying mucosal disruption, lamina propria disorganization, and increased collagen deposition. DOCI imaging revealed that uninjured tissue displayed shorter lifetimes (blue–green spectrum) and injured tissue showed longer lifetimes (yellow–red spectrum). Quantitative analysis confirmed a statistically significant difference between injured and uninjured groups. These findings suggest that DOCI can detect biochemical and structural changes to reliably differentiate scarred and normal tissue, showing strong potential for noninvasive early detection, monitoring, and reduced reliance on invasive biopsy.

Air pollution remains one of the leading threats to human health, contributing to the development of cardiovascular disease, respiratory conditions, and other inflammatory responses within the human body. Pollutants stemming from biomass burning, including woodsmoke particles (WSP), contribute significantly to the global health burden due to their role in promoting oxidative stress and inflammatory responses. While previous studies have demonstrated how WSP disrupt iron homeostasis in epithelial cells, their effects on macrophages remain poorly understood. This study investigates how RAW 264.7 macrophages exposed to water-soluble woodsmoke particle extracts (WSP-WS) and humic-like substances (HULIS) affect labile iron concentrations and iron homeostasis pathways. Macrophages were exposed to WSP-WS or HULIS (500 µg/mL), with or without co-treatment using ferric ammonium citrate (FAC) or diphenyl oxide (DPO). Labile iron (Fe2+) was quantified using a FerroOrange stain with subsequent fluorescence microscopy, with intensity normalized to the brightfield signal and image area. We determined that HULIS exposure significantly increases labile iron levels, suggesting a disruption of iron homeostasis and elevated importer activity. This finding is consistent with a mechanism in which HULIS sequesters intracellular iron, triggering uptake of external iron and an accumulation in the labile iron pool. Co-treatment with FAC or DPO mitigated these effects, thus restoring iron homeostasis, returning iron concentrations to baseline levels.