Effect of type lll collagen coating of electrospun scaffolds on breast cancer cell apoptosis

Breast cancer arises from the epithelial or the connective tissue components of the breast. Breast cancer is the most commonly diagnosed cancer in women, with about half a million women dying because of the disease each year. Local recurrence (return of cancer in the area of primary diagnosis) and metastasis (spread of cancer to other locations in the body) are major challenges associated with breast cancer, leading to significant mortality and morbidity in patients. Although death is due to metastasis, local recurrence is a gateway towards metastasis. Therefore, there arises a need for targeted therapeutic treatments to treat local recurrence and ultimately metastasis in breast cancer patients. In this project, we have developed novel, electrospun scaffolds to function as a therapeutic strategy for breast cancer, based on the wound healingfibrosis- cancer progression triad (WHFC). Collagen deposition and remodeling command a pivotal role in the WHFC. Previous work from the Volk Lab (University of Pennsylvania) supports a role for Collagen type lll (Col 3) in the WHFC triad. Notably, their data show that Col 3 deficiency promotes tumor growth and metastases, promotes proliferation, inhibits apoptosis of breast cancer cells, and fosters a procarcinogenic stroma by regulating collagen, myofibroblast density, and alignment. Based on this data and other research which shows that post-surgical wound healing response in breast resection/biopsy sites drives aggressive tumor behaviors, we hypothesized that application of Col 3 coated biomaterials to breast cancer resection sites would potentially promote an optimal wound healing response, would suppress aggressive breast cancer behavior, and invoke a proapoptotic response in residual breast cancer cells. By being able to do so, such biomaterials would reduce morbidity and mortality in breast cancer patients by preventing local recurrence and breast cancer metastasis. Additionally, this non-toxic approach would decrease morbidity by decreasing the need for more toxic adjuvant therapies such as radiation or chemotherapy that are currently used to treat breast cancer. Biomaterials have played an important role in improving quality of life for breast cancer patients by maintaining cosmesis in the treated breast and, therefore, a biomaterial that could improve healing and aesthetic outcome while controlling tumor behavior would provide a unique advantage to those commercially available. In the scope of this project, we focused our investigation on in vitro assessment of the ability of Col 3 coated electrospun (fibrous) biomaterials to induce apoptosis of a human triple negative breast cancer (TNBC) cell line (MDA-MB-231). In this investigation, we determined if the architectural makeup of these fibrous scaffolds, in terms of fiber orientation, would have a further effect on breast cancer cell morphology and ultimately on breast cancer cell apoptosis. Lastly, we assessed if Col 3 coated substrates could induce apoptosis in other cancer cell types. To initially assess apoptosis, MDA-MB-231s were seeded onto coverslips (1.13 cm2 surface area; 48 hours) coated with equal amounts of diluted recombinant Col 3 (1 [mu]g/cm2) and Col 1 (1 [mu]g/cm2). Apoptosis was induced with 1 [mu]M Staurosporine (24 hours). MDA-MB-231 cells cultured on Col 3 coated cover slips exhibited a significantly higher level of apoptosis compared seeded cells to the fiber orientation (possibly through limiting contact guidance). Based on these experiments, we decided to check if unaligned PCL scaffolds would potentiate Col 3 induced apoptosis of breast cancer cell line. It is possible that the alteration of normal mechanotransduction in elongated (associated with aggressive phenotype) cancer cells, where they convert mechanical signals to biochemical signals, would affect their apoptotic ability. To test this hypothesis, we conducted preliminary comparative studies on MDA-MB-231 cells on Col 3 coated aligned and unaligned PCL scaffolds and found that Col 3 coated unaligned PCL scaffolds resulted in a significantly higher apoptosis than Col 3 coated-aligned PCL scaffolds (p=0.0066). Finally, we hypothesized that Col 3-based therapies might be effective in controlling other cancer types. To test this hypothesis, we examined if Col 3 could induce significant apoptosis in other cancer cell lines. Col 3-coated cover slips promoted significantly higher levels of apoptosis (p=0.0392) in human lung carcinoma cell line A549, than those seeded onto Col 1- coated cover slips (control). Therefore, our data suggests that Col 3-coated fibrous biomaterials with an unaligned fiber orientation significantly promote breast cancer cell apoptosis, thereby providing us with a novel and safe method to potentially control clinical outcomes in breast cancer patients. Our results also suggest that these Col 3-containing substrates should be further investigated for use in controlling additional (non-breast) cancers.