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Date of Award

Spring 5-15-2018

Author's School

Graduate School of Arts and Sciences

Author's Department

Biology & Biomedical Sciences (Molecular Cell Biology)

Degree Name

Doctor of Philosophy (PhD)

Degree Type

Dissertation

Abstract

Breast cancer is the most common cancer for women worldwide, representing approximately 25% of all new cancer cases in this population. While early detection and removal of breast cancer still confined to the primary site results in a good prognosis, approximately one- third of patients will develop distant metastases. In these patients, overall survival is markedly reduced. Of the common sites for breast cancer metastasis, the skeletal system is the most frequent. Treating breast cancer bone metastases has proven particularly difficult for several reasons, such as dissemination of metastases throughout the skeleton, poor drug localization to sites of interest, a lack of tumor-specific targets expressed across breast cancer subtypes, and the chemo-protective nature of the bone microenvironment. This dissertation is focused on investigating a potential tumor-target expressed on breast cancer bone metastases, and to improve drug treatment efficacy against tumor cells in the bone microenvironment.

Integrins are heterodimeric cell surface receptors, composed of an α and β subunit from a large family of selectively-compatible integrin subunits. As a heterodimeric complex, integrins can bind to components of the extracellular matrix or to other cells. One particular integrin complex, integrin αvβ3, is composed of the tightly regulated integrin subunit β3 and the more widely expressed αv subunit. I examined the expression of integrin β3 on primary breast cancer as compared to metastases in murine cancer models, and observed that integrin expression is significantly elevated on bone metastases as compared to the primary tumors or visceral metastases. In addition, I evaluated tumor-associated integrin β3 expression on a tissue microarray (TMA) composed of primary breast cancer and patient-matched bone metastatic tissue from 42 patients. Across nearly all patients, tumor-associated integrin β3 expression was significantly elevated on bone metastases as compared to the primary tumor. For the first time, I demonstrate that tumor-associated integrin β3 is elevated on bone metastases across all breast cancer subtypes, supporting the translational potential of targeting integrin β3 in breast cancer patients with bone metastases.

Integrin β3 was weakly expressed on tumor cells in vitro and on tumor cells in the primary mammary fat pad (MFP). Additional analysis demonstrated that integrin β3 on circulating tumor cells is dispensable for strong expression of integrin β3 on subsequent bone metastases, suggested that integrin β3 may be induced within the bone microenvironment. I identified transforming growth factor beta (TGF-β) to be a potent inducer of integrin β3 in vitro, and further demonstrate canonical TGF-β signaling through the SMAD2 and SMAD3 (SMAD2/3) pathway is responsible for breast cancer upregulation of integrin β3 induction on bone metastases, both in vitro and in vivo.

Utilizing this information, I sought to evaluate the targeting potential of nanotherapy coated with a targeting ligand specific for integrin αvβ3. Nanotherapy has the potential to increase therapeutic efficacy and reduce toxicity versus traditional chemotherapies by enhancing drug delivery to specific targets of interest. I explored the localization potential of two nanoparticles with significantly different sizes: polysorbate (tween) 80 micelle nanoparticles (MPs, ~12.5 nm) or perfluorocarbon (PFC) nanoparticles (~250 nm). The smaller integrin αvβ3- targeted micelle nanoparticle (αvβ3-MP) could more effectively penetrate breast cancer bone metastases than larger integrin αvβ3-targeted PFC nanoparticles (αvβ3-PFCs). With these observations, I evaluated whether αvβ3-MP-mediated drug delivery could more effectively attenuate bone metastatic tumor burden and bone destruction than free drug delivery.

Using the chemotherapeutic agent docetaxel (DTX), a potent microtubule inhibitor that is a first-line therapy for metastatic breast cancer, I observe that DTX is only weakly tumor- suppressive in our mouse model of breast cancer metastases. However, treating mice bearing breast cancer metastases with αvβ3-MP-delivery of a docetaxel-prodrug (DTX-PD) significantly reduced bone tumor burden and bone destruction, and with less hepatotoxicity. I observed a significant decrease in bone-residing tumor cell proliferation in mice treated with αvβ3-MP- delivery of DTX-PD, without overt osteoclast killing or inhibition of osteoclast formation. Together, these results provide support that nanotherapy-mediated attenuation of bone metastases and bone destruction occurs through enhanced drug efficacy against breast cancer cells within the bone.

In this Dissertation, Chapter 1 will provide an overview of breast cancer, bone metastases, integrins, and the therapeutic potential of nanotherapy. In Chapter 2, my work on the expression and physiologic regulation of integrin β3 on breast cancer during metastases will be explored. In Chapter 3, the role of the cytokine TGF-β in regulating tumoral expression of integrin β3 will be discussed. And in Chapter 4, I discuss the use of integrin αvβ3-targeted nanotherapy directed against breast cancer metastases. Collectively, I provide evidence that chemotherapeutic efficacy against breast cancer cells within bone can be enhanced by exploiting the expression of tumoral integrin β3 at that metastatic site.

Language

English (en)

Chair and Committee

Katherine Weilbaecher

Committee Members

Greg Lanza, Daniel Link, Fanxin Long, Deborah Veis,

Comments

Permanent URL: https://doi.org/10.7936/K7QR4WK7

Available for download on Sunday, May 15, 2118

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