Harrison, RogerWoodward, Alexis2022-12-122022-12-122022-12-16https://shareok.org/handle/11244/336921Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subset with limited targeted chemotherapies. TNBC is defined by the absence of the estrogen, progesterone, and HER2 hormonal receptors on the cell surface, and targeted hormonal-based therapies are ineffective. The current standard of care for these patients remains untargeted systemic chemotherapy that does not discriminate between healthy and cancerous tissue, resulting in severe dose-limiting side effects. Identifying a novel biomarker for this disease remains a top priority in targeted cancer therapy. Extracellular exposure of phosphatidylserine (PS) is tightly regulated in healthy cells. Typically confined to the inner cytosolic leaflet, once extracellularly exposed, it acts as an “eat me” signal for macrophage phagocytosis. However, cancer (especially TNBC) cells have high externalized PS but do not undergo apoptosis, making PS a promising biomarker for TNBC therapy. Annexin A5 (ANXA5) is the native binding partner to PS and can be used to actively target and deliver chemotherapeutic agents to the tumor microenvironment via PS externalization. While many advancements have been made in developing targeted chemotherapeutics, protein-drug conjugates have changed how cancer is treated. Protein-drug conjugates consist of a protein linked to a drug, and depending on the conjugation method, a linker that connects the protein and drug together. The protein acts as a delivery vehicle to localize a drug to cancer cells and vasculature through a cancer biomarker. By targeting and localizing the drug to the tumor, less drug is needed to induce cell death, resulting in fewer side effects. This dissertation will focus on translating two ANXA5 protein drug conjugates and developing one ANXA5-protein drug conjugate for the treatment of TNBC. ANXA5 has proven to be an invaluable delivery vehicle with high specificity to externalized PS. A microtubule inhibitor, a DNA alkylating agent, and a histone deacetylase inhibitor have all been linked to ANXA5, and all three ANXA5 drug conjugates have shown better anticancer activity than their respective free drug in vitro. ANXA5 protein drug conjugates provide a novel avenue to treat an untargeted cancer.Annexin A5Protein Drug ConjugatesImmunogenic Cell Death