Fusobacterium nucleatum (Fn) is a commensal bacterium and opportunistic pathogen associated with multiple diseases, including oral squamous cell carcinoma (OSCC). Despite the prevalence and aggressiveness of OSCC, the specific Fn factors that contribute to OSCC progression remain largely unknown. Identifying these bacterial determinants is essential for understanding microbe-cancer interactions and developing targeted therapeutic strategies. To address this, a genome-wide functional screen was performed using a sequence-defined Fn Tn5 transposon mutant library. By utilizing transposon mutagenesis, approximately 1,400 unique Fn mutants were generated. This library was screened in a GFP-expressing OSCC cell line using a spheroid model, allowing assessment of cancer growth following bacterial infection. Mutants that significantly altered spheroid growth were subsequently validated through secondary assays that evaluated spheroid formation, spheroid expansion, and 2D and 3D cell migration. This pipeline identified two Fn genes critical in Fn-mediated OSCC progression. These findings demonstrate that discrete Fn genetic factors can directly influence OSCC growth and invasion-related properties. This study achieves two aims: identifying specific Fn determinants that influence OSCC progression, and establishing a framework for systematic discovery of Fn virulence factors involved in OSCC. Future studies will focus on characterization of these factors and their downstream effects to evaluate their potential as therapeutic targets.

The gastrointestinal (GI) tract is considered to be the largest endocrine organ in the human body. While many studies have established that the large intestine microbiota regulates distally produced gut hormones such as serotonin and GLP-1, little is known about the effects of the small intestine microbiota on proximally produced gut hormones. Cholecystokinin (CCK), a gut hormone best known for its role in gastric emptying, pancreatic enzyme secretion, and satiety, is primarily produced in the duodenum of the small intestine by I-cells, a subtype of specialized endocrine cells in the GI tract. While nutrient sensing is the primary established trigger for CCK release by I-cells, recent studies identify receptors for microbially derived signals on I-cells, suggesting a potential relationship between small intestine microbes and CCK. Here, we demonstrate that small intestine microbiota play a critical role in regulating host CCK bioavailability. Using a gnotobiotic mouse model without a microbiome, we found that mice without a microbiome have low levels of CCK, and this was restored upon transplantation of small intestine microbiota but not fecal microbiota. Altogether, these findings highlight a distinct role for small intestine microbiota in modulating small intestine hormones with implications for metabolic and gut-brain health.

Alzheimer’s Disease (AD) is a pervasive and difficult disease with limited options for therapy. One key feature of AD pathogenesis is the aggregation of beta-amyloid (Aβ) peptides into Aβ plaques, which interfere with neuronal communication and degrade brain function. Chimeric Antigen Receptor (CAR) therapy has been revolutionary in many disease therapies, and CAR-Macrophages (CAR-Ms) can be especially useful to fight AD as they utilize phagocytosis to engulf their target. However, current immunotherapies with anti-Aβ antibodies (Donanemab & Lecanemab) are hindered by potential excess neuroinflammation (NI). Our project aims to engineer and test Hematopoietic Stem Cell (HSC)-derived CAR-Ms with Aβ-binding regions and an NI-reducing gene, TREM2, incorporated to target Aβ plaques in AD. TREM2 is a native receptor for Aβ known to reduce NI upon signaling, and loss is associated with increased AD pathology. We found that HSCs transduced with TREM2 lentivirus can successfully engraft and generate TREM2-overexpressing macrophages in a mouse model of AD, 5XFAD. We have also successfully cloned and produced a CAR-TREM2 lentiviral vector that expresses both the Aβ CAR and TREM2. In the future, the optimized CAR-HSCs will be injected into 5XFAD mice. Plaque clearance, cognitive improvement, and CNS inflammation will be measured. We anticipate that our optimized CAR-M will successfully remove Aβ plaques while reducing NI, and that mice treated with this CAR-M will show lower neurodegeneration, leading to an effective treatment of AD.

Chronic lung allograft dysfunction (CLAD) remains the leading cause of long-term mortality following lung transplantation, driven by persistent alloimmune activation. While emerging hypoimmune cell engineering strategies aim to mitigate rejection through evading the immune system, these approaches have not yet proven effective for solid organ transplantation, necessitating continued reliance on pharmacologic immunosuppression. The JAK/STAT pathway is a central mediator of cytokine-driven immune activation, but current systemic JAK inhibitors are associated with significant toxicity, highlighting the need for targeted alternatives. This study seeks to investigate TD3014, a novel pan-JAK inhibitor designed for regional delivery, using a bone marrow-derived dendritic cell (BMDC)–splenocyte mixed lymphocyte reaction (MLR) model that recapitulates alloimmune responses observed in CLAD. Gene expression of Th1, Th2, and Th17 pathways was quantified via RT-qPCR, and it was discovered that the BMDC-MLR model exhibited a dominant Th1 response, with strong upregulation of IFN-γ and NOS2, alongside moderate Th2 and variable Th17 activation. TD3014 treatment suppressed inflammatory gene expression across all pathways, returning levels to baseline rather than ablating signaling. These findings suggest TD3014 enables controlled immunomodulation, preserving basal immune function while limiting pathological alloimmunity, supporting its potential as a targeted therapy for CLAD.

Toxoplasma gondii is the obligate intracellular parasite responsible for toxoplasmosis, a chronic disease that infects roughly a third of the world’s population. In T. gondii, the apicoplast is a non-photosynthetic organelle responsible for essential metabolic pathways and is particularly attractive as a potential drug target due to its specificity to apicomplexan parasites. Apicoplast biogenesis and segregation are still not fully understood and all the molecular factors involved have yet to be characterized. This study identifies and studies several factors that play critical roles in this process. Recently, we discovered apicoplast partitioning protein 1 (APP1), a novel protein that causes apicoplast inheritance defects when disrupted. Subsequently, through proximity labeling (TurboID) of APP1, TgGT1_313360 was identified as a potential interactor. TgGT1_313360 was endogenously tagged and found to localize as punctate structures on the apicoplast, with accumulation near the centrosomes. Disruption of this protein resulted in apicoplast inheritance defects similar to those of APP1, thus we named it APP2. Investigations of the relationship between APP1 and APP2 through immunofluorescence analyses revealed that APP1 associated with the cytoskeleton, whereas APP2 was relegated to the apicoplast. Altogether, these findings expand our understanding of apicoplast segregation machinery and open new avenues of exploration of this essential process in the parasite.