We live in an age of ever-pressing environmental crises. Scientists have determined that the cumulative impact of human activity on Earth in the last 300 years reaches a scale equal to that of a geological epoch—carving marks in the planet that will last longer than our species has existed. Practices of extraction—mining for coal, drilling for oil—are one collection of activities integral to that impact, prompting scholars to investigate the complex of cultural factors that drive and ultimately constitute these practices. Regarding these practices, scholars in the environmental humanities have raised an important question: how, in the first place, does nature get figured as a resource? How has Western humanity’s relation towards the natural world developed into one predominantly of exploitation? In this paper, I investigate the metaphysical basis of this relationship through Martin Heidegger’s ‘method’ of reflection [Besinnung]. My investigation begins with the thought of Descartes and Bacon, two philosophers who, at the commencement of modernity, interpreted the human as the relational center of being as such. This early-modern, anthropocentric philosophy prefigures an event Heidegger calls gestell—the disclosure of all beings exclusively as resources—which I argue is the underlying metaphysical basis of practices of extraction. I go on to argue that the very proliferation of extractive thinking reveals the crucial importance of poetry and philosophy today, both of which embody a kind of thinking that is antithetical to extraction and instrumentalization, a thinking that—rather than dominating, organizing, utilizing—lets things show themselves as they are. I conclude that art and philosophy both counter the ubiquitous logic of extraction and are therefore invaluable to any attempt at figuring a fuller, healthier mode of humanity’s relating to itself and the natural world.

My research analyzes the portrayal of curses as symbolic literary devices reflecting socioeconomic anxieties in Shakespeare’s Macbeth and Joseph Conrad’s Nostromo. These texts serve as a lens to explore contemporary institutional failings, the rise of capitalism, and the impact of empire on resource-rich communities. Spanning nearly three centuries, this comparison highlights the persistent tragedy in human struggles with power and belief systems. Macbeth depicts a literal curse, under the cover of prophecy, leading to a relentless and bloody pursuit of power, and the central character's eventual downfall. Nostromo, in contrast, adopts a metaphorical stance on curses, with characters seeking absolution from their imported belief systems. The connection between these works lies in the evolution of cursed institutions – from the early belief systems in governance depicted in Macbeth to their manifestation as global capitalist structures in Nostromo. This study explores how these literary curses symbolize the tragic, cyclical nature of power and societal upheaval, mirroring modern global anxieties and the pursuit of institutional absolution. Employing a tragic thematic analysis, I integrate contemporary studies of current systems and institutions with historical source examination. Alongside a close reading of both texts, this method enables a nuanced understanding of the characters’ actions and their broader impact. My research aims to provide fresh perspectives on Macbeth and Nostromo, highlighting how magical language in both texts serves as a tool for understanding and navigating complex global challenges. It seeks to uncover insights into how literary curses and prophecies may mirror, and potentially forecast, power dynamics, societal disruptions, and ecological issues, offering novel interpretations of our current socioeconomic and political landscape.

Although immunotherapy with T-cells is successful in treating non-solid cancers, targeting solid cancer tumors remains a challenge. Unlike T-cells, bacteria can colonize solid tumors and thrive in a hypoxic tumor microenvironment (TME). This means that bacteria could be used to treat solid tumors that T-cells cannot reach. I propose to develop new bacterial immunotherapies that can be used as an alternative treatment method to fight solid cancer tumors. Specifically, I aim to engineer E. coli that secretes therapeutic payloads upon sensing the TME. My research focused on comparing secretion efficiencies of signal peptides and secretion tags. Signal peptides are short sequences that transport cargoes, such as therapeutic proteins, to the periplasm. Similarly, secretion tags are small secreted proteins that can transport a partner cargo fused to them to the extracellular medium. From literature, I selected the signal peptide PelB and the secretion tag YebF. I compared their efficiencies in secreting the cargo human interleukin 2 (h-IL2), an immunostimulatory cytokine. To this end, I expressed h-IL2 with either the genes for the signal peptide or secretion tag fused to the N-terminus and a detection tag on the C-terminus, in an E. coli expression strain. I induced the expression of cargoes, after which I isolated the proteins that were secreted into the extracellular medium. I detected the proteins through quantitative Western blot analysis. I concluded from my experimental data that the cargoes were secreted at a higher concentration with YebF than with PelB. I plan to repeat this experiment with another secretion tag, OsmY. The next step is to use the secretion system with the highest secretion yield to secrete a variety of potential immunomodulatory cargoes. I plan to evaluate their effects on immune signaling and their ability to eliminate tumor cells.

Genome-wide association studies demonstrate an association between the single nucleotide polymorphism (SNP) rs3184504 and many autoimmune diseases, including Type 1 Diabetes (T1D). The rs3184504*C allele encoding for arginine is mutated to encode for tryptophan in the rs3184504*T risk allele, resulting in a loss-of-function of the adaptor protein SH2B3, a negative regulator of various tyrosine kinases and cytokine receptors. The SNP is uniquely enriched in humans of European descent. Isolation of the SNP rs3184504 is challenging due to its presence in a linkage disequilibrium region. To normalize this donor variability we used a CRISPR/Cas9 system in human peripheral blood mononuclear cells (PBMCs). CRISPR guides target exon 4 of the SH2B3 gene to be cut by the Cas9 enzyme. This cut is then repaired by a green fluorescent (GFP) repair template cassette delivered by an adeno-associated virus (AAV) containing homology arms for the flanking SH2B3 sequence. The insertion of GFP permits the identification of the SH2B3 KO cells. Our previous experiments on SH2B3 KO mouse lines demonstrated heightened signaling response after IL-2 stimulation in T cells, particularly in CD8+ naive and effector cells and CD4+ effector cells. To test this in primary human T cells, we will stimulate our edited SH2B3 KO cells with IL-2 and use flow cytometry to compare differential responses between edited and unedited T-cell populations. We expect to see similar results in our edited cells. Successful gene editing of PBMCs to model the human SH2B3 disease haplotype enables us to more accurately study the biological underpinnings of T1D, ultimately providing new avenues for which to treat T1D.

The human genome is associated with numerous variations, some of which can be pathogenic. Any variations larger than 50 base pairs (bp) are considered structural variants (SVs), and SVs impacting copy number of the genome are termed copy number variants (CNVs). By studying CNVs, we can understand the underlying mechanisms of damage and repair within DNA, which can help work towards prevention and cure in other fields, such as cancer genomics. My project investigates a CNV on chromosome 2 of a pediatric patient, who presents with tetralogy of Fallot, global development delay, and multiple other congenital anomalies. Using array comparative genomics hybridization (aCGH), we identified a 3.2 megabase (Mb) complex genomic rearrangement (CGR) spanning the 2q31 region of the proband’s chromosome 2. Detailed analysis of the short-read whole genome sequencing (WGS) allowed us to locate the exact coordinates of each junction, or the beginning and end points of each extra copy. PacBio long-read genome sequencing and optical genome mapping detected the same junctions and facilitated confirmation of the overall structure. The CGR can be characterized as a duplication-triplication-duplication-triplication-duplication (DUP-TRP-DUP-TRP-DUP), meaning this segment of the genome contains a segment of alternating 1 and 2 extra copies of this region. The CGR is de novo, or not inherited from the proband’s parents. Due to the nature of this variant, it is likely to be impacting the phenotype of our patient. To establish a genotype-phenotype correlation, I did a literature review of patients with overlapping CGRs, comparing their phenotypes to our proband, as well as reviewing any known disease-associated genes in the region using the online catalog of human genes and genetic disorders (OMIM). Patient and disease comparison revealed the extreme rarity of our patient’s CGR, leading us to believe the phenotype of the proband results from impact of multiple genes in the affected region.

Merkel cell carcinoma (MCC) is a rare and aggressive cancer of the skin with a mortality rate of ~30%. In the US, most MCC tumors arise from integration of the Merkel cell polyomavirus (MCPyV) DNA into a host chromosome, leading to expression of viral T-Antigen (T-Ag) oncoproteins that drive tumorigenesis. Though current treatment options have significantly improved MCC prognosis, new therapies are needed to address recurrent/resistant disease. While T-Ag-specific antibodies are usually detected in the blood of patients with virus-driven MCC, the role of these antibodies in tumor immunity remains unclear. Here, we analyzed blood samples from 100 MCC patients prior to definitive treatment, 51 of whom had high titers of antibodies recognizing the T-Ags. These 51 high titer samples were assessed for binding across two domains of the T-Ag. Suprisingly, we found that patients who had high titers of antibodies binding both regions of the T-Ag had worse MCC control than patients whose antibodies predominantly bound one region (median PFS 5.5 vs. 14.2 months, p=0.003). These data suggest that careful mapping of circulating antibody reactivity to different regions of T-Ag can serve as a biomarker to identify high-risk patients for which a more aggressive treatment regimen is needed. Future work is also focused on understanding the immune response resulting in differential response to T-Ag domains.

Stress-related physiological and social factors can have a direct impact on cancer biology and patient outcomes. Exposure to a stressor can lead to downstream activation of signaling pathways that can impact cancer-related processes. Compared to older and younger patients with cancer, adolescents and young adults (AYAs) have had less improvement in clinical outcomes and report high levels of psychosocial distress. The goal of this study is to examine the feasibility and acceptability of a multimethod triangulation protocol collecting stress biomarkers along with subjective patient reported outcomes among AYAs with cancer. Eligible participants are 12-24 years old within six months of initial cancer diagnosis and undergoing treatment at Seattle Children's Hospital. Once enrolled, participants wear a sensor measuring autonomic nervous system activity [quantified as heart rate variability (HRV)] while completing validated patient reported outcome (PRO) surveys assessing psychosocial symptoms and participating in a qualitative interview querying the same domains. Interviews are recorded using Microsoft Office Dictation services. I listen to the audio files and go over the transcribed document making corrections and highlighting key quotations. Additionally, at the time of a participant's planned clinical blood dram, 5mL of blood is collected to measure a social genomics stress biomarker called the conserved transcriptional response to adversity (CTRA). We expect to find greater than 70% feasibility and 80% acceptability in this study. Feasibility is defined as the percentage of participants who complete all (100%) study procedures; acceptability is defined as the percentage of participants who agree that similar patients could complete the study. Results from this pilot study will inform future longitudinal investigations and the development of targeted biopsychosocial interventions for AYAs with cancer.