Date of Graduation

5-2025

Document Type

Dissertation (PhD)

Program Affiliation

Cancer Biology

Degree Name

Doctor of Philosophy (PhD)

Advisor/Committee Chair

Scott Kopetz

Committee Member

Giulio Draetta

Committee Member

David Menter

Committee Member

Kristin Eckel-Mahan

Committee Member

Anirban Maitra

Committee Member

Giannicola Genovese

Committee Member

Michelle Barton

Abstract

Colorectal cancer minimal residual disease (MRD) represents a major clinical problem for colorectal cancer patients, with failure rates of surgery and adjuvant chemotherapy between 5% to 40% depending on stage of disease. In our study, we simulated MRD using genetically engineered organoids with precise somatic editing of APC and TP53, creating a murine model that mimics human liver metastatic colorectal cancer. By implementing a meticulously timed experimental metastatic model, we could detect microscopic tumor lesions. Through genomic and transcriptomic analyses, we pinpointed the importance of macrophages, particularly those expressing high levels of CSF1R, in these microscopic metastatic focal lesions. We conducted macrophage- targeted immunotherapy using anti-CSF1R. Treatment of micro-metastases resulted in complete remission of microscopic disease and eradication of macrophages expressing CD163+ after four weeks of treatment. However, anti-CSF1R treatment did not exhibit efficacy in treating macro-metastases. Spatial transcriptomic analysis, along with multiplex immunofluorescent analysis, demonstrated the effectiveness of anti-CSF1R treatment in significantly reducing macrophage populations and enhancing CD8+ T cells after 14 days of treatment. Anti-CSF1R holds promise as a targeted therapy for colorectal cancer MRD patients.

Keywords

Colorectal cancer, Tumor immune microenvironment, Immunology, Genetics, Immunotherapy, Genetic engineering mouse models

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