Date of Award
2025
Document Type
Dissertation
Degree Name
Doctor of Philosophy (PhD)
Department
Cellular, Molecular and Biomedical Sciences
First Advisor
Brian Cunniff
Abstract
Breast cancer remains a leading cause of cancer related mortality worldwide, with triple-negative breast cancer (TNBC) representing one of the most aggressive and therapeutically challenging subtypes. Mitochondrial positioning within cells has emerged as a critical determinant of cancer cell metabolism, signaling, and survival, yet the molecular mechanisms governing these spatial dynamics and their contribution to tumorigenesis are not well understood. Miro1 (mitochondrial Rho GTPase 1) is a protein embedded in the mitochondrial outer membrane. Miro1 is essential for coordinating mitochondrial trafficking along microtubules through its interactions with adaptor proteins such as TRAK1/2 and motor proteins. Beyond its canonical role in mitochondrial transport, Miro1 is also involved in calcium homeostasis, organelle contact site formation, and mitophagy. Importantly, published data indicate that breast cancer patients whose tumors express high levels of Miro1 have significantly poorer survival outcomes compared to those with low Miro1 expression, highlighting the importance of investigating its role in the molecular mechanisms that drive breast cancer progression.In recent years, Miro1 has been implicated in supporting cellular processes such as migration and proliferation in both normal and malignant cells, including breast cancer. To investigate the role of Miro1 in breast cancer tumorigenesis and metastasis, we generated MDA-MB-231 human breast cancer cells with stable knockdown of Miro1. Loss of Miro1 led to increased phosphorylation of ERK1/2, loss of DUSP6 translation, and increased 4EHP expression. In our functional assays, Miro1 knockdown was associated with reduced cell proliferation, migration, and invasion in vitro. In vivo, xenografts derived from Miro1 knockdown cells resulted in significantly reduced tumorigenesis compared to wild-type controls. To further assess the role of Miro1 in mammary tumor development in vivo, we generated a novel transgenic mouse model with inducible, tissue-specific deletion of Miro1 in mammary epithelial cells, in combination with polyomavirus middle T-antigen (PyVMT) oncogene activation. In this model, Miro1 wild-type mice developed tumors at all mammary gland sites and exhibited pulmonary metastases. In contrast, mice with complete Miro1 deletion failed to form tumors despite oncogene activation. Notably, heterozygous mice with only one intact Miro1 allele developed primary tumors but displayed a markedly reduced capacity for metastasis. Together, these findings demonstrate that Miro1 is not merely a regulator of mitochondrial positioning, but a key modulator of cellular behaviors that drive both the initiation and progression of breast tumors. Its loss impairs primary tumor formation, disrupts oncogenic signaling pathways, and diminishes the metastatic potential of breast cancer cells. The observed dose-dependent effects ranging from complete tumor suppression in Miro1-null mice to reduced metastasis in heterozygous models further emphasize the importance of Miro1 expression levels in determining tumor initiation. These data suggest that Miro1 may serve as a valuable biomarker for tumor status, reflecting both proliferative capacity and metastatic risk. Moreover, the findings highlight Miro1 as a promising therapeutic target, where modulation of mitochondrial positioning and associated signaling pathways could offer novel strategies for treating aggressive subtypes such as triple-negative breast cancer.
Language
en
Number of Pages
184 p.
Recommended Citation
Termos, Randi, "Miro1-Mediated Mitochondrial Positioning In The Development And Metastasis Of Breast Cancer" (2025). Graduate College Dissertations and Theses. 2166.
https://scholarworks.uvm.edu/graddis/2166