Lung adenocarcinoma (LUAD) in females is shaped by both immune microenvironment dynamics and tumor cell differentiation. Alpha-ketoglutarate (αKG), a key metabolite in the tricarboxylic acid cycle, has previously been shown to influence stem cell pluripotency and immune responses. Here, we investigated whether dietary calcium-αKG could slow LUAD progression in female mice by enhancing differentiation and modifying immune infiltration. A genetically engineered LUAD mouse model was generated by intranasal AdCre delivery at 12 weeks of age, followed by 16 weeks of 2% calcium-αKG supplementation in the standard mouse chow diet. Hematoxylin & eosin staining revealed that αKG-treated tumors were significantly smaller. Bulk RNA sequencing demonstrated activation of a differentiation-associated transcriptional program, including MYOGENESIS, and increased expression of the transcription factor TBX5. Western blot analysis further revealed a reduction in the repressive histone modifications H3K27me3 and H3K9me3, consistent with an open chromatin state. Interestingly, immunohistochemistry revealed that αKG-treated mice showed reduced CD3⁺ T cell infiltration without changes in CD68⁺ macrophage populations. RNA sequencing analysis confirmed that αKG decreased expression of immune checkpoint genes in female LUAD tumors. Overall, these results suggest that dietary αKG reprograms LUAD tumors in females by promoting differentiation and altering immune contexture through epigenetic remodeling, offering potential insight into metabolism-based therapeutic strategies.    

Lung cancer is a leading cause of cancer-related deaths in the United States and worldwide. Non-small cell lung cancer (NSCLC) makes up 85% of all lung cancers, with the most common subtypes being adenocarcinoma and squamous cell carcinoma. In adenocarcinomas, the epidermal growth factor (EGFR), a single-pass transmembrane receptor tyrosine kinase (RTK) which regulates many important cellular processes, is mutated in approximately 15-20% of patients. Gain-of-function mutations in EGFR are commonly observed; several are known to be oncogenic. The high frequency of EGFR mutations has led to the generation of EGFR tyrosine kinase inhibitors (TKIs); however, resistance frequently emerges. Most EGFR-mutant lung cancer patients present with primary mutations that can be targeted with EGFR TKIs, but after prolonged treatment, secondary mutations can arise causing inhibitor resistance. We employed functional genetic screens identifying secondary mutations causing resistance to osimertinib, a third-generation TKI, in distinct EGFR mutant backgrounds: exon 19 deletion and L858R, the two most common EGFR primary mutations. We expressed top enriched EGFR secondary mutations in each background and assessed their ability to drive resistance in the PC9 lung cancer cell line using colony formation and western blot assays. The colony formation assay showed subsets of secondary mutations in the L858R background drive resistance, while others drive resistance in both the exon 19 deletion and L858R. The western blot showed increased levels of phosphorylated-ERK in resistance-causing mutations treated with osimertinib, correlating with increased colony formation. The results suggest EGFR secondary mutations, in both backgrounds, drive resistance to osimertinib through ERK-signaling.    

Pancreatitis is a progressive fibroinflammatory disease of the pancreas, mainly characterized by visceral pain. In the most severe cases this pain may be relieved by total pancreatectomy with islet autotransplantation (TPIAT). Repeated visceral pain exposure can sensitize peripheral and central nerves, creating neuropathic pain that presents as hyperalgesia or allodynia. Quantitative Sensory Testing (QST) characterizes pain mechanisms and assesses altered pain processing. We studied neurological pain regulation using QST in pancreatitis patients undergoing TPIAT. 20 patients (mean age 36 years, 25% male) in a multicenter study (POST) underwent pre-TPIAT QST; 7 completed repeat testing 1–2 years post-surgery. Nineteen age- and sex-matched healthy controls were enrolled. QST measured pain detection thresholds to pressure and thermal stimuli, and ice water immersion tested conditioned pain modulation. 1 year TPIAT outcomes (pain, quality of life (QoL)) were collected. We compared pre-TPIAT patients to control QST results, pre- to post-TPIAT changes, and investigated whether pre-TPIAT QST predicts surgical outcomes. TPIAT and healthy control participants had similar pain detection thresholds. In the subset with follow-up, thresholds increased after TPIAT, suggesting reduced hyperalgesia, while CPM responses appeared to dampen. Associations of preoperative QST measures with QoL outcomes are still under investigation. Preliminary results from this pilot study suggest limited neuropathic pain in TPIAT recipients, compared to healthy volunteers. Despite the small sample, findings indicate neurologic changes in pain detection thresholds after visceral pain removal achieved through TPIAT. Ongoing investigations will further clarify associations between sensory testing, neuropathic pain mechanisms, and surgical outcomes for TPIAT in pancreatitis patients.

The resurgence of Mpox virus (MPVX), a zoonotic orthopoxvirus, has raised significant public health concerns due to its increasing human-to-human transmissibility and the current lack of effective antiviral therapies. This study aims to identify novel drug targets by systematically cloning and functionally characterizing previously unstudied or poorly annotated MPXV genes. Using the pENTR/D-TOPO vector with TOP10 Chemically Competent E. coli cells, we have successfully cloned and sequence verified over 180 MPVX genes. Optimized techniques, including ethanol precipitation and primer redesign, have significantly improved plasmid purity and sequencing accuracy, ensuring high-quality constructs for downstream applications. These genes will be expressed in bacterial and mammalian systems to assess protein localization and interactions relevant to viral replication and immune evasion. Computational assays, such as Alpha folding models, are being used to visualize the protein and its potential role in the viral process. Subsequent drug screening efforts will evaluate potential inhibitors targeting these proteins in cell-based models to assess their potential to disrupt viral lifecycle processes. Through cloning the entire genome, we can provide foundational tools for MPVX drug discovery. The findings contribute to broader efforts in virology and support the development of targeted therapeutic strategies essential for future outbreak preparedness.  

The actin cytoskeleton is a dynamic structure that is required for cellular motility, trafficking, and force generation. Formins, such as Formin Homology 2 Domain-Containing 3 (FHOD3), promote actin nucleation (the formation of new filaments) and elongation (the extension of existing filaments). The Formin Homology 2 (FH2) Domain controls filament nucleation. In humans, mutations in FHOD3 account for 1-2% of cases of hypertrophic cardiomyopathy (HCM), a condition that affects 1 in 500 individuals (Ochoa et al., 2018). To test how FHOD3 mutations impact actin assembly activity, and thus could cause disease, we use fluorescent in vitro actin assembly assays and TIRF microscopy. Unpublished data from our lab show that one HCM-associated mutation in FH2 leads to a 37% increase in nucleation ability in vitro. However, no study has carefully examined other likely pathogenic mutations in the FH2 domain. I plan to investigate two other FH2 HCM mutations, S1138N and Y1424N, and determine how they impact FHOD3 actin assembly. We expect to see an increase in nucleation rate, consistent with the previously studied HCM mutation. These experiments will analyze how these mutations affect nucleation rate, shedding light on FH2’s role in dynamic actin regulation. While this study is currently ongoing, the results are expected to align with previous work and lead to further clarification on how these pathogenic mutations alter the function of FHOD3 and contribute to the sarcomere disorganization present in HCM.