Brian W. Booth

Photodynamic therapy (PDT) is a minimally invasive cancer treatment method. The antitumor effect of PDT mediated by the photosensitizer meta-tetra(hydroxyphenyl)chlorin (m-THPC) was studied in vitro on HER2-positive (HER2⁺) breast cancer cells. Since the application of m-THPC is limited due to poor tumor targeting abilities, the polymer carrier dextran-grafted-polyacrylamide (D-g-PAA) with attached gold nanoparticles was loaded with m-THPC to provide a light-responsive, tumor targeting nanocomposite. The nanocomposite was absorbed by the cancer cells within 90 min and activated through irradiation with red light at a wavelength of 660 nm. In this study, the efficiency and ideal dose of the nanocomposite were evaluated and determined based on cell viability and induction of apoptosis of HER2⁺ breast cancer cells and human breast epithelial cells. We demonstrate that the nanocomposite exhibited phototoxicity with cell mortality exceeding 50% for the HER2⁺ breast cancer cells. In contrast, the human breast epithelial cells were minimally affected at the same concentration of the nanocomposite, demonstrating the tumor-targeting abilities of the developed nanocomposite. Dark toxicity was also comparably lower, indicating the limited effect of the unactivated nanocomposite. The results revealed that apoptosis was the primary mode of cell death in HER2⁺ breast cancer cells. Apoptotic cell death was induced in HER2⁺ breast cancer cells via the caspase-3, caspase-9, and Bax pathways. In conclusion, the studied nanocomposite demonstrated excellent PDT performance with good antitumor activity under light irradiation in vitro and is worthy of further investigation as an alternative treatment for HER2⁺ breast cancer.

Chemotherapy is commonly used to treat patients with triple-negative breast cancer. Combinations of platinum-based chemotherapies have demonstrated higher rates of pathologic complete responses of triple-negative breast cancer compared to combinations without platinum-based chemotherapies. However, there is a significant increase in general toxicity with the addition of platinum-based regimens. Some groups have investigated using tumor-treating fields as an alternative treatment method for triple-negative breast cancer, and chemosensitization by tumor-treating fields has been observed in vitro. With the goal of minimizing the toxicities associated with platinum-based chemotherapy, we investigated anti-mitotic effects of concurrent tumor-treating fields and cisplatin treatment to show that the addition of tumor-treating fields further inhibited cell growth in triple-negative breast cancer. We show that combining cisplatin with tumor-treating fields induces higher levels of apoptosis within triple-negative breast cancer cells as opposed to normal epithelial cells and the combination has a more immediate mechanism of cell death than either monotherapy. We also demonstrate that, when combined with tumor-treating fields, a lower dose of chemotherapy can be used to achieve the same efficacy of triple-negative breast cancer cell death as higher doses of individual therapy.

Photodynamic therapy (PDT) is a non-invasive anticancer treatment that uses special photosensitizer molecules (PS) to generate singlet oxygen and other reactive oxygen species (ROS) in a tissue under excitation with red or infrared light. Though the method has been known for decades, it has become more popular recently with the development of new efficient organic dyes and LED light sources. Here we introduce a ternary nanocomposite: water-soluble star-like polymer/gold nanoparticles (AuNP)/temoporfin PS, which can be considered as a third-generation PDT system. AuNPs were synthesized in situ inside the polymer molecules, and the latter were then loaded with PS molecules in an aqueous solution. The applied method of synthesis allows precise control of the size and architecture of polymer nanoparticles as well as the concentration of the components. Dynamic light scattering confirmed the formation of isolated particles (120 nm diameter) with AuNPs and PS molecules incorporated inside the polymer shell. Absorption and photoluminescence spectroscopies revealed optimal concentrations of the components that can simultaneously reduce the side effects of dark toxicity and enhance singlet oxygen generation to increase cancer cell mortality. Here, we report on the optical properties of the system and detailed mechanisms of the observed enhancement of the phototherapeutic effect. Combinations of organic dyes with gold nanoparticles allow significant enhancement of the effect of ROS generation due to surface plasmonic resonance in the latter, while the application of a biocompatible star-like polymer vehicle with a dextran core and anionic polyacrylamide arms allows better local integration of the components and targeted delivery of the PS molecules to cancer cells. In this study, we demonstrate, as proof of concept, a successful application of the developed PDT system for in vitro treatment of triple-negative breast cancer cells under irradiation with a low-power LED lamp (660 nm). We consider the developed nanocomposite to be a promising PDT system for application to other types of cancer.

Purpose: Currently, tumor-treating field (TTField) therapy utilizes a single “optimal” frequency of electric fields to achieve maximal cell death in a targeted population of cells. However, because of differences in cell size, shape, and ploidy during mitosis, optimal electric field characteristics for universal maximal cell death may not exist. This study investigated the anti-mitotic effects of modulating electric field frequency as opposed to utilizing uniform electric fields. Methods: We developed and validated a custom device that delivers a wide variety of electric field and treatment parameters including frequency modulation. We investigated the efficacy of frequency modulating tumor-treating fields on triple-negative breast cancer cells compared to human breast epithelial cells. Results: We show that frequency-modulated (FM) TTFields are as selective at treating triple-negative breast cancer (TNBC) as uniform TTFields while having a greater efficacy for combating TNBC cell growth. TTField treatment at a mean frequency of 150 kHz with a frequency range of ± 10 kHz induced apoptosis in a greater number of TNBC cells after 24 h as compared to unmodulated treatment which led to further decreased cell viability after 48 h. Furthermore, all TNBC cells died after 72 h of FM treatment while cells that received unmodulated treatment were able to recover to cell number equivalent to the control. Conclusion: TTFields were highly efficacious against TNBC growth, FM TTFields showed minimal effects on epithelial cells similar to unmodulated treatment.

Purpose: Triple-negative breast cancer continues to be one of the leading causes of death in women, making up 7% of all cancer deaths. Tumor-treating electric fields are low-energy, low-frequency oscillating electric fields that induce an anti-proliferative effect on mitotic cells in glioblastoma multiforme, non-small cell lung cancer, and ovarian cancer. Little is known about effects of tumor-treating fields on triple-negative breast cancer and known research for tumor-treating fields only utilizes low (< 3 V/cm) electric field intensities. Methods: We have developed an in-house field delivery device capable of high levels of customization to explore a much wider variety of electric field and treatment parameters. Furthermore, we investigated the selectivity of tumor-treating field treatment between triple-negative breast cancer and human breast epithelial cells. Results: Tumor-treating fields show greatest efficacy against triple-negative breast cancer cell lines between 1 and 3 V/cm electric field intensities while having little effect on epithelial cells. Conclusion: These results provide a clear therapeutic window for tumor-treating field delivery to triple-negative breast cancer.

The tumor microenvironment is recognized as performing a critical role in tumor initiation, progression, and metastasis of many cancers, including breast cancer. The breast cancer microenvironment is a complex mixture of cells consisting of tumor cells, immune cells, fibroblasts, and vascular cells, as well as noncellular components, such as extracellular matrix and soluble products. The interactions between the tumor cells and the tumor microenvironment modulate tumor behavior and affect the responses of cancer patients to therapies. The interactions between tumor cells and the surrounding environment can include direct cell-to-cell contact or through intercellular signals over short and long distances. The intricate functions of the tumor microenvironment in breast cancer have led to increased research into the tumor microenvironment as a possible therapeutic target of breast cancer. Though expanded research has shown the clear importance of the tumor microenvironment, there is little focus on how normal mammary epithelial cells can affect breast cancer cells. Previous studies have shown the normal breast microenvironment can manipulate non-mammary stem cells and tumor-derived cancer stem cells to participate in normal mammary gland development. The tumorigenic cells lose their tumor-forming capacity and are “redirected” to divide into “normal”, non-tumorigenic cells. This cellular behavior is “cancer cell redirection”. This review will summarize the current literature on cancer cell redirection and the normal mammary microenvironment’s influence on breast cancer cells.

Introduction: Breast cancer research has advanced understanding of breast cancer development and progression greatly over the past few decades. Despite progress in research, breast cancer is the second leading cause of cancer death in North America and is the most frequent type of cancer for women. Better treatments and diagnostic tools have increased breast cancer survival rates, but there is still a fundamental lack of understanding in tumor progression. Previous studies show the normal mammary microenvironment can influence non-mammary cells and tumor-derived cancer cells to participate in normal mammary gland development. The tumorigenic cells lose their tumor-forming capabilities and are “redirected” into phenotypically normal, non-tumorigenic cells. The purpose of this study is to obtain knowledge of the mechanisms that play a role in cancer cell redirection and therefore be able manipulate those mechanisms for therapeutic treatment in a clinical setting. Our hypothesis is that microenvironmental elements control whether a tumorigenic cell will enter a redirected state. We have developed and validated an in vitro model to mimic the mammary microenvironment to study cancer cell redirection. We found that when cancer cells that overexpress HER2+ are redirected in our cancer redirection model, phospho-HER2+ is silenced. We use HER2+ phosphorylation as a marker for cancer cell redirection though not a mechanism for cancer cell redirection. Materials and Methods: HER2+ breast cancer cells and normal breast epithelial cells (BECs) were co-cultured in ratios of 1:1 or 1:50. Monocultures of breast cancer cells and BECs were used as controls. Western analysis and immunostaining was used to assess attenuation of HER2⁺ and phospho-HER2⁺ receptors and RNAseq was used for pathway analyses. Images were taken using a Leica confocal microscope. For comparison of 2 or more groups, a one-way analysis of variance was performed. A p-value of less than 0.05 was considered significant. Results and Discussion: We found that the redirected cells underwent a phenotype shift in which the redirected cells adopted a normal mammary epithelial phenotype based on gene expression profiles. Furthermore, when HER2+ breast cancer cells were redirected in vitro they lost their tumor-forming potential in vivo. Signal pathway analyses revealed that redirected cancer cells are adopting a normal phenotype compared to breast cancer cells. Conclusions: These results indicate that epithelial cells provide signals that influence HER2+ breast cancer cells to undergo a shift in phenotype. The phenotypic shift in cancer cell redirection includes multiple intracellular signaling pathways that may be the key towards effective cancer treatment. Acknowledgements: This research was supported by South Carolina Idea Networks of Biomedical Research Excellence (SC INBRE). Citation Format: Caroline J. Campbell, Brian W. Booth. Investigating the mechanisms of HER2+ breast cancer cell redirection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2536.

Tannic Acid (TA) is a naturally occurring antioxidant polyphenol that has gained popularity over the past decade in the field of biomedical research for its unique biochemical properties. Tannic acid, typically extracted from oak tree galls, has been used in many important historical applications. TA is a key component in vegetable tanning of leather, iron gall ink, red wines, and as a traditional medicine to treat a variety of maladies. The basis of TA utility is derived from its many hydroxyl groups and its affinity for forming hydrogen bonds with proteins and other biomolecules. Today, the study of TA has led to the development of many new pharmaceutical and biomedical applications. TA has been shown to reduce inflammation as an antioxidant, act as an antibiotic in common pathogenic bacterium, and induce apoptosis in several cancer types. TA has also displayed antiviral and antifungal activity. At certain concentrations, TA can be used to treat gastrointestinal disorders such as hemorrhoids and diarrhea, severe burns, and protect against neurodegenerative diseases. TA has also been utilized in biomaterials research as a natural crosslinking agent to improve mechanical properties of natural and synthetic hydrogels and polymers, while also imparting anti-inflammatory, antibacterial, and anticancer activity to the materials. TA has also been used to develop thin film coatings and nanoparticles for drug delivery. In all, TA is fascinating molecule with a wide variety of potential uses in pharmaceuticals, biomaterials applications, and drug delivery strategies.

The development of multifunctional biomaterials as both tissue regeneration and drug delivery devices is currently a major focus in biomedical research. Tannic Acid (TA), a naturally occurring plant polyphenol, displays unique medicinal abilities as an antioxidant, an antibiotic, and as an anticancer agent. TA has applications in biomaterials acting as a crosslinker in polymer hydrogels improving thermal stability and mechanical properties. We have developed injectable cell seeded collagen beads crosslinked with TA for breast reconstruction and anticancer activity following lumpectomy. This study determined the longevity of the bead implants by establishing a degradation time line and TA release profile in vivo. Beads crosslinked with 0.1% TA and 1% TA were compared to observe the differences in TA concentration on degradation and release. We found collagen/TA beads degrade at similar rates in vivo, yet are resistant to complete degradation after 16 weeks. TA is released over time in vivo through diffusion and cellular activity. Changes in mechanical properties in collagen/TA beads before implantation to after 8 weeks in vivo also indicate loss of TA over a longer period of time. Elastic moduli decreased uniformly in both 0.1% and 1% TA beads. This study establishes that collagen/TA materials can act as a drug delivery system, rapidly releasing TA within the first week following implantation. However, the beads retain TA long term allowing them to resist degradation and remain in situ acting as a cell scaffold and tissue filler. This confirms its potential use as an anticancer and minimally invasive breast reconstructive device following lumpectomy.