Yes-associated protein (YAP1) is a transcriptional activator that targets cell development and proliferation genes. YAP1 is known to have oncogenic activity in breast cancer development. When overactive within the cell, nuclear translocation of YAP1 is upregulated, leading to increased transcription and cell growth. YAP1 nuclear import is facilitated by karyopherin subunit alpha 1 (KPNA1). However, the mechanisms of YAP1 nuclear translocation are unclear, as YAP1 lacks a canonical nuclear localization signal (NLS), which is typically required for KPNA1 binding. The purpose of this study is to determine how YAP1 interacts with KPNA1 to allow for nuclear import. We have expressed and purified a construct of YAP1 containing the WW domains required for KPNA1 binding, as well as a full length construct of a murine karyopherin subunit alpha. We plan to co-crystallize a complex of these two constructs, and use X-ray crystallography to solve their structure. After identifying key interactions at the binding interface, site directed mutagenesis will be performed to verify the proposed complex. Characterization of the YAP1 nuclear translocation pathway may inform its potential as a target for future drug discovery efforts in the breast cancer field. Furthermore, it may provide insight into the nuclear import of other molecules lacking a canonical NLS.

Adaptive decision-making requires evaluating cost-benefit tradeoffs to select optimal actions. The anterior cingulate cortex (ACC) is connected to the dorsomedial striatum (DMS), and this circuit monitors action choices based on the value of the expected outcome. Disruption of this pathway impairs the ability to update effortful choices in response to shifting reward contingencies, leading to maladaptive decision-making in conditions such as depression, schizophrenia, and ADHD. Our goal was to determine how ACC projections to DMS contribute to effort-based choice behavior, specifically whether they encode the need to shift toward high-effort strategies when such choices become more valuable. Mice performed an automated barrier T-maze task, choosing between a low-effort/low-reward path and a high-effort/high-reward path requiring climbing a 15 cm barrier. We virally expressed GCaMP in ACC→DMS terminals and monitored calcium with fiber photometry during behavior. Following high-reward outcomes with a barrier present, optic fibre recordings indicated that ACC-DMS activity was suppressed. In contrast, suppression was reduced after low-reward outcomes, indicating that the reward was integrated. These findings highlight the ACC-DMS pathway as a key node for integrating effort–reward information. Future work will investigate how reward signals evolve as mice adapt their choice strategies, as well as utilize optogenetics for a better understanding of ACC-DMS dynamics.

The sex hormones estradiol and testosterone, while mainly associated with reproduction, are involved in other processes outside of reproduction, and especially in the regulation of whole-body energy balance   through the brain. Previous research has discovered a sex difference present in the feeding behavior regulated by somatostatin (SST) neurons in the Tuberal Nucleus (TN) of the hypothalamus, in which ablation decreases food intake only in lean females with high estrogens. In this study, we aimed to dissociate the roles of testicular and ovarian sex hormones in the regulation of food intake via TN-SST neurons. We hypothesized that TN-SST neurons integrate metabolic and ovarian hormones to regulate food intake . To study the influence of gonadal hormones on TN-SST neurons, male and female Sst-Cre mice received gonadectomies (GDX) or sham surgeries, in combination with injection of AAV-Caspase, which ablates TN-SST neurons, or AAV-GFP as a control. We found that lean females experienced an increase in food intake after a GDX. Furthermore, the loss of testicular hormones caused an expected decrease in food intake. Surprisingly, ablation rescued this effect in GDX males, indicating that testicular hormones also regulate feeding behaviors through TN-SST neurons. These results indicate that both ovarian and testicular hormones are involved in the regulation of feeding by TN-SST neurons. Understanding the influence of sex hormones on feeding behavior may assist in the development of treatments for conditions in which metabolism and sex interact, such as obesity, polycystic ovarian syndrome (PCOS), and changes in weight during menopause and aging across sexes.

Lung cancer remains the leading cause of cancer-related mortality worldwide, and incidence and mortality rates are higher in males. Alpha-ketoglutarate (αKG), a tricarboxylic acid cycle metabolite with roles in epigenetic regulation, has been implicated in tumor biology, but its in vivo effects in lung adenocarcinoma (LUAD) remain unclear. To investigate, a genetically engineered male LUAD mouse model was fed a 2% calcium-αKG diet for four months after AdCre induction, followed by histological, molecular, and imaging analyses. αKG supplementation was associated with increased tumor area, greater adiposity, and weight gain. Gene set enrichment analysis revealed activation of inflammatory pathways and enrichment of epithelial-to-mesenchymal transition signatures. RNA sequencing indicated immune remodeling, with upregulation of immunosuppressive markers and downregulation of immune checkpoint genes. Immunohistochemistry confirmed reduced CD3⁺ T cell infiltration and increased CD68⁺ macrophages, suggesting a shift toward tumor-associated macrophage dominance. Epigenetically, histone analysis demonstrated elevated H3K27me3 in male LUAD tumors, suggesting enhanced repressive chromatin states. MRI and microCT confirmed increased visceral adiposity in αKG-fed mice. Together, these findings demonstrate that dietary αKG exacerbates tumor progression in a male LUAD model through interconnected metabolic, inflammatory, and epigenetic mechanisms, highlighting the need to further dissect sex-specific responses to metabolite supplementation.

Clear cell renal cell carcinoma (ccRCC) is the most common form of kidney cancer in adults, with over 40% of cases progressing to metastasis. Metastatic renal cell carcinoma (mRCC) is highly aggressive and often fatal. Between 2014 and 2020, patients with localized disease had a 93% survival rate, while those with metastatic disease had only 18%. Despite this disparity, few treatments exist for the metastatic stage. ccRCC is commonly driven by loss of function of the Von Hippel-Lindau (VHL) tumor suppressor gene, leading to increased expression of Hypoxia-Inducible Factors (either HIF1A or HIF2A). Our lab recently discovered that VHL-negative, HIF1A-positive ccRCC cells likely drive metastasis within heterogeneous tumors containing HIF2A-positive cells. High-throughput drug screens revealed that several statins selectively killed these VHL-negative, HIF1A-positive cells. Preliminary mouse studies further showed that fluvastatin treatment inhibited kidney cancer lung metastasis. My project focuses on validating this selective toxicity using clonogenic assays on genetically engineered human ccRCC cell lines. Results show that human ccRCC cells are highly sensitive to statins, supporting their potential as therapeutic agents. Next, I will test whether fluvastatin can inhibit metastasis in a human ccRCC tumor model containing both VHL-negative, HIF1A-positive and HIF2A-positive cells. If successful, these studies will corroborate our murine findings and confirm that VHL-negative, HIF1A-positive cells drive metastasis. Future work will investigate the HIF1A-dependent anti-metastatic mechanism of statins. Given that statins are already FDA-approved to lower cholesterol, repurposing them to prevent metastatic kidney cancer could offer a safe, cost-effective, and life-saving therapeutic strategy.