Pulmonary hypertension secondary to pulmonary fibrosis (PF-PH) is classified as Group 3 pulmonary hypertension and remains relatively underinvestigated. The prevalence PF-PH has increased over time and is associated with right ventricular (RV) failure. We hypothesize that PF-PH is characterized by increased vascular remodeling and inflammation. Ten male Sprague-Dawley rats were divided into control (n = 5) and PF-PH (n = 5) groups. PF-PH was induced using intratracheal bleomycin (to induce PF) followed by subcutaneous monocrotaline (to induce PH) two weeks later. Serial echo and terminal cath were performed to assess the hemodynamics. At the end of five weeks the rats were sacrificed, and lungs were stained with pentachrome to assess vascular remodeling. Average wall thickness and lumen size were quantified using ImageJ, and a statistical comparison was performed using two tailed t-test. RNA was isolated from the lungs and RNA sequencing and bioinformatic analysis were performed. PF-PH rats developed severe PH as demonstrated by increased RV systolic pressure and Fulton index. Lung sections from PF-PH rats demonstrated increased pulmonary vascular wall thickness. RNA sequencing demonstrated significant upregulation of pathways and genes related to vascular remodeling and inflammation. In conclusion, PF-PH is characterized by increased RV pressure, hypertrophy, vascular remodeling, and inflammation. Future studies are warranted to investigate the role of specific genes related to inflammation and vascular remodeling in PF

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous pollutants widely distributed in commercial and industrial products. Legacy PFAS have been associated with cancer, immune dysfunction, and metabolic disruption. Safer PFAS alternatives such as GenX, specifically hexafluoropropylene oxide dimer acid, have replaced legacy PFAS due to their shorter half-life; however, evidence suggests GenX may exhibit similar toxicological profiles as legacy PFAS, including potential immunotoxicity. Therefore, we sought to evaluate the stain- and sex-specific immunomodulatory effects of GenX. Male and female mice from two genetically distinct mouse strains, C57BL/6J and DBA/2J (DBA), were exposed to GenX over 6 weeks. As a result, exposure to GenX increased pro-inflammatory cytokines across all groups, with DBA female mice showing stronger effects. To evaluate the molecular mechanisms of GenX on DBA female mice, we conducted single-cell RNA sequencing on the respective blood leukocytes. We found that T cells displayed the largest transcriptomic shift followed by B cells, natural killer cells, macrophages, dendritic cells, and neutrophils. Pathway enrichment analysis identified TNF-alpha Signaling as broadly affected across cell types. We also observed cell type specific responses like KRAS Signaling Up in neutrophils and E2F Targets in T cells. These results demonstrate the impact of GenX on specific cell types and pathways in the immune system, leading to a greater understanding of its potential role in immune dysregulation.

Regenerative medicine involves using therapeutic cells that can repair damaged cells and tissue. Extracellular vesicles are lipid bilayer particles secreted by all cells as a mode of communication. Cell therapy induces tissue repair by secreting EVs rich in bioactive small RNAs. Still, little is understood about the function of these small RNAs and uncovering their mechanism can inspire the development of novel RNA drugs. In previous work, we identified yREX3, an EV-derived small Y RNA that enhances the ability of immune cells (namely macrophages) to engulf and clear material. Here we report the development of TY2, a synthetic small RNA bioinspired by yREX3. Methods. TY2 is a synthetic and modified RNA drug bioinspired by yREX3. TY2 demonstrated enhanced stability and therapeutic equivalence compared to yREX3. We evaluated TY2 in a mouse model of sepsis produced by cecal-ligation and puncture (CLP). Results. Mice given repeated intravenous injections of TY2 showed improved cardiac function, reduced systemic inflammation and reduced bacterial burden in tissue. In an in vitro assay, TY2 enhances uptake of inactivated E. Coli by mouse bone marrow-derived macrophages. Conclusions. Thus, TY2 is a novel bioinspired small RNA drug with therapeutic potential. More broadly, these findings rationalize this novel EV RNA-based paradigm for drug discovery.

High grade gliomas (HGG) are malignant, aggressive cancers that are characterized by altered metabolism, including increased reliance on aerobic glycolysis and lactate production, which supports tumor growth and survival. This study sought to evaluate targeting monocarboxylate transporter 1 (MCT1), a critical regulator of lactate transport, in a pediatric high-grade glioma (pHGG) cell line with elevated MCT1 expression. This project studied the effects of the MCT1 inhibitor AZD3965 on tumor growth and lactate metabolism in the PBT-05 pHGG cell line. The initial hypothesis was that AZD3965 would be effective in inhibiting tumor growth by blocking MCT1’s role in the export of lactate out of tumor cells, which aids in supporting tumor growth. Growth inhibition was assessed using a CellTiter-Glo ATP viability assay and cell imaging via Incucyte, while extracellular lactate accumulation was measured using a Lactate-Glo assay. Results demonstrated that AZD3965 reduced ATP production with an IC50 of 3.191 nM and decreased lactate export with an IC50 of 0.712 nM. Incucyte analysis showed that AZD3965 reduced tumor growth by approximately 50% at physiologically relevant concentrations. These findings suggest that MCT1 inhibition with AZD3965 can reduce ATP production, limit tumor growth, and reduce lactate export in PBT-05 pHGG cells, demonstrating its potential as a therapeutic drug. Future research should explore therapies with other molecules to enhance tumor-killing effect, and evaluate drug potency in an in vivo animal system.

Statins are a class of small molecule, FDA approved drugs that are widely used to combat cardiovascular disease due to their cholesterol lowering effect. Previous results from our lab indicate that statins, specifically older statins such as fluvastatin and pitavastatin, demonstrate selective cytotoxicity towards highly metastatic cancer cell lines but not their less aggressive counterparts. This pattern was first noted in kidney cancer but has since been replicated in breast cancer and prostate cancer. My project focuses on the selectivity of statins on neuroendocrine prostate cancer (NEPC) and seeks greater clarity on the currently unknown mechanism of action (MOA) involved in statins’ anti-cancer effects. I will be examining the role of hypoxia pathways with emphasis on two proteins, hypoxia inducible factors 1 alpha and 2 alpha (HIF-1a & HIF-2a). This work follows data from our lab’s kidney cancer project, which suggests HIF-1a as the likely target of statins and the hypoxia program as integral to our understanding of a MOA. First, I aim to characterize the expression of these two proteins via western blot analysis across our nine established prostate cancer cell lines with degrees of severity ranging from adenocarcinoma to NEPC. Second, I will attempt to generate HIF-1a and HIF-2a knockouts of our most stable NEPC cell line for future statin dose-response curves, which will ideally reveal the role of these proteins in statins’ cytotoxicity.