Alzheimer’s Disease (AD) is a progressive brain disorder that debilitates memory, learning, and decision-making. Early-stage AD represents the initial phase where individuals are still able to function independently, but with increasing age, their condition steadily progresses to dementia and loss of independence. Because a significant number of the aging population is affected by AD, understanding the neuroinflammatory processes would help develop more effective strategies for treatment. Examining markers such as MCP-1 and TNF-alpha, known to be associated with inflammatory response, will help identify the modulatory processes that lead to mild cognitive impairment associated with early-stage AD. Subsequently, higher levels of inflammation markers within the brain leads to mild cognitive impairment. This research study involved 40 C57BL/6 mice, 20 males and 20 females (21 months old), retro-orbitally infected with 80 µL of neurotrophic AAV-AD vector or AAV-Sham for a duration of 2 months before humane euthanasia. Brains were collected, and specific regions were examined by immunohistochemistry (IHC) and digital imaging to assess the expression levels and distribution of the inflammation markers. Preliminary observations showed that hippocampal regions of the brain from mice with early-stage AD had higher staining intensity for MCP-1and TNF-alpha compared to respective areas in Sham mice, suggesting increased inflammation is a very early lesion that develops in the presence of AD pathogenic components that might be controlled by anti-inflammatory drugs. The preliminary data suggests that the characteristics of AD manifest in part due to the neuroinflammatory response of brain factors that change with onset AD.

Stimulator of Interferon Genes (STING) signaling contributes to tumor immunity. However, treatments targeting the STING pathway are limited by route of administration, insufficient STING activation, and off-target toxicity. We introduce poly-STING, a copolymerized, mannosylated variant of the diABZI STING agonist-3 known to activate the cGAS-STING signaling pathway, promoting the release of type-1 interferons and pro-inflammatory cytokines leading to tumor immunogenicity. The STING agonist-3 is a non-nucleotide molecule that activates the STING pathway, but it has poor solubility, which limits its usage in-vivo. The developed poly-STING platform improves the drug's solubility, is designed to target immune cells, and provides enzyme-triggered drug release upon delivery, which has been shown to induce improved therapeutic efficacy compared to the free drug. The Pun and Stayton labs seek to investigate modalities for optimization of the cGAS-STING pathway activation and characterize the mechanism of action. Specifically, my project will evaluate STING activation by observing macrophage repolarization from type M2, as the mannose from the poly-STING binds to the CD206 receptors on M2 macrophages. This activates the STING pathway, repolarizing the macrophage to pro-inflammatory type M1. To test effects in vitro, I will culture bone marrow-derived M2 macrophages with various formulations of poly-STING, and repolarization will be measured through flow cytometry and RT-qPCR to quantify expression of macrophage markers. We expect to find higher M1 activity in macrophages treated with poly-STING as opposed to the free drug. Next, I evaluate the therapeutic efficacy of the STING formulations through an in-vivo tumor reduction study using murine models of breast cancer and melanoma, expecting to find longer survival of mice treated with poly-STING. The culmination of this project will result in a polymer-based STING agonist delivery platform that solves the solubility and bioavailability issues associated with the STING-3 agonist, with enhanced efficacy and decreased toxicity after systemic administration.

Merkel cell carcinoma (MCC), a rare and aggressive skin cancer, shows approximately 60% response to immune-checkpoint inhibitors (ICIs) in immunocompetent patients. The efficacy of ICIs in immunosuppressed patients, who generally have poorer MCC prognoses, is less clear. This retrospective study assesses ICI outcomes in both immunocompetent and immunosuppressed MCC patients, and across various immunosuppression types. In this project, I determined the cohort of 183 patients with advanced MCC who were treated with first-line ICIs from a Seattle-based data registry. I collected the following data from analyzing patients’ medical records: treatment response, immunosuppressive status, disease-specific and overall survival. I coordinated with the statisticians for Kaplan-Meier analyses of the data, and am now working on the manuscript. The results show that initial response rates to ICIs were comparable between immunosuppressed (50%) and immunocompetent (61.5%) patients (p=0.17). After starting ICI treatment, immunosuppressed patients experienced a 65% increased rate of disease progression compared to immunocompetent patients (Hazard Ratio [HR]=1.65, p=0.04), with their median time to disease progression being 11.2 months, versus 32.9 months in the immunocompetent group. Outcomes also varied by immunosuppression subtype, with chronic lymphocytic leukemia patients having the lowest chance of response (20%, 2/10) and highest progression risk. In our cohort, immunotherapy often elicited an initial response in immunosuppressed MCC patients; however, the duration of this response was significantly shorter. Despite limited duration in immunosuppressed patients, ICIs still offer a high response rate for patients with advanced MCC, regardless of immunosuppression type. It is crucial to have a comprehensive prognosis discussion with patients before initiating treatment.

Heschl’s gyrus (HG) is a region of the brain containing the primary auditory cortex. The extent of folding within the HG shows high morphological variability across individuals. Interestingly, increased HG folding is more likely to be found in expert than amateur musicians, suggesting a possible role of auditory experience in shaping HG gyrification. In my research, I examined HG folding in blind individuals—another population with extensive auditory experience. I hypothesized that, if experience alters HG structure, then individuals with early-onset blindness might have increased HG gyrification compared to those with late-onset blindness or those who are sighted. I analyzed T1-weighted images collected from previous MRI studies at the University of Washington, University of Pennsylvania, and Oxford University. The combined dataset included 6 anophthalmia (individuals born without eyes), 48 early blind, 18 late blind, and 28 sighted control participants. I created hand-drawn HG regions of interest for each participant in both hemispheres and measured HG gyrification in two ways: 1) by visually categorizing the extent of HG folding (single, partial, or complete duplication), and 2) by obtaining continuous metrics (gyrification index and curvedness index) using FreeSurfer. A chi-squared test revealed that the degree of HG folding was not different across the four groups. A linear mixed-effects model (controlling for the effects of age, hemisphere, and scan location), similarly showed no effects of group on the gyrification index or the curvedness index. To conclude, my findings show that blindness does not affect HG gyrification. The results challenge the idea that auditory experience alters HG structure and offer important insights into previous findings in professional musicians. Our results suggest that the prevalence of duplicated HG in musicians may be the result of individuals with larger processing capacity within the auditory cortex being more likely to take up music as a profession.