Graduation Year

2017

Document Type

Dissertation

Degree

Ph.D.

Degree Name

Doctor of Philosophy (Ph.D.)

Degree Granting Department

Biology (Cell Biology, Microbiology, Molecular Biology)

Major Professor

Conor C. Lynch, Ph.D.

Committee Member

Wenlong Bai, Ph.D.

Committee Member

Srikumar P. Chellappan, Ph.D.

Committee Member

John M. Koomen, Ph.D.

Keywords

PTHrP, IGF, Osteoblast, Osteoclast, skeletal malignancy, MMP

Abstract

Skeletal metastasis is a lethal component of many advanced cancers including prostate, the second most common cancer among men. Patients whose prostate cancer is localized and detected early benefit from multiple treatment options ranging from active surveillance to radiation and surgery, resulting in a 5-year survival rate of nearly 100%. Unfortunately, the prognosis and survival for patients with advanced metastatic disease is much worse due to the highly aggressive nature of the disease and a paucity of treatment options. Understanding the mechanisms and interactions that occur between metastatic cancer cells and the bone will enable the future treatment landscape for bone metastatic prostate cancer to expand, thereby improving patient outcomes. Our current knowledge of how metastatic prostate cancer cells interact with the bone is summarized in a model known as the “vicious cycle.” Numerous fundamental vicious cycle factors have been identified, including parathyroid hormone-related protein (PTHrP), while additional elements, such as matrix metalloproteinases (MMPs), are progressively being discovered and added to the model.

PTHrP is a critical regulator of bone resorption and augments osteolysis in skeletal malignancies. In Chapter 2, we report that the mature PTHrP1-36 hormone is processed by MMPs to yield a stable product, PTHrP1-17. PTHrP1-17 retains the ability to signal through PTH1R to induce calcium flux and ERK phosphorylation but not cyclic AMP production or CREB phosphorylation. Notably, PTHrP1-17 promotes osteoblast migration and mineralization in vitro, and systemic administration of PTHrP1-17 augments ectopic bone formation in vivo. Further, in contrast to PTHrP1-36, PTHrP1-17 does not affect osteoclast formation/function in vitro or in vivo. Finally, immunoprecipitation-mass spectrometry analyses using PTHrP1-17-specific antibodies establish that PTHrP1-17 is indeed generated by cancer cells. Thus, MMP-directed processing of PTHrP disables the osteolytic functions of the mature hormone to promote osteogenesis, indicating important roles for this mechanism in bone remodeling in normal and disease contexts.

MMPs have traditionally been associated with cancer progression based on their extracellular matrix degrading activities. However, it has become evident that their regulation of non-extracellular matrix substrates can exert both contributive and protective effects during tumorigenesis. Previous studies of matrix metalloproteinase-3 (MMP-3) have demonstrated tissue dependent pro- and anti-tumorigenic effects, but despite elevated expression, its roles have not been explored in bone metastatic prostate cancer. In Chapter 3, we show that tumor-derived MMP-3 contributes to prostate tumor growth in bone. In vitro, we observe that silencing MMP-3 reduces prostate cancer cell proliferation. Further, we found increased levels of IGFBP3, a known MMP-3 substrate, and decreased IGF-1R, ERK, and AKT phosphorylation in the MMP-3 silenced cells. Notably, we also observe reduced tumor growth and proliferation in in vivo intratibial models when tumor-derived MMP-3 expression is silenced. These data suggest that increased MMP-3 expression by prostate cancer cells contributes to their proliferation in bone by regulating the activity of the IGF/IGF-1R signaling axis.

Taken together, our studies indicate that MMPs possess important functional roles in bone metastatic prostate cancer. We believe that elucidation of these mechanisms and their contributions to the vicious cycle of bone metastasis will offer novel opportunities to design effective therapeutic treatment options.

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