According to U.S. Breast Cancer Statistics, about 1 in 8 U.S. women will develop invasive breast cancer during their lifetime. Chemotherapeutics that are used on patients currently often lead to tumor resistance, bone marrow suppression and cachexia. This study evaluated a novel combination of three non-mutagenic compounds for their effectiveness against mammary tumor cells, toxicity towards immune cells, ability to provoke the expression of immunogenic cell death (ICD) markers, and killing in 3D tumor models. Methotrexate (MTX), 2-deoxyglucose (2DG), and wogonin (WGN) were combined at doses well below their EC50 values yet effectively killed human and mouse breast cancer cells. The combination inhibited cancer cell colony formation and induced a high degree of cell death in multiple malignant tumor cell lines. Importantly, the combination did not significantly inhibit the viability of peripheral-blood mononuclear cells (PBMCs), even when employed at 3X the concentration that killed cancer cells. In marked contrast, low-dose doxorubicin, a common therapeutic for breast cancers, significantly decreased PBMC viability and increased the percentage of cell death. Our novel combinatorial therapy (Trifecta) elicited the significant expression of three ICD hallmarks: calreticulin surface expression, ATP secretion, and HMGB-1 release. In all cases, Trifecta elicited an equal or greater degree of ICD-marker expression compared to doxorubicin, a known inducer of ICD. We show significant efficacy of Trifecta against human and mouse mammary 3D tumor models grown in Matrigel® ECM-complex containing culture medium, and reaffirm the marked resistance of tumorspheres towards the conventional chemotherapeutic doxorubicin. The effectiveness of Trifecta in an acceptable surrogate model for mouse studies bodes well for translation of our findings to the clinic. In conclusion, Trifecta has proven highly effective against tumor cells grown either as monolayers or tumorspheres, without significant cytotoxic effects towards proliferating immune cells. Furthermore, treatment with this combination elicits ICD, which has the potential to prime an adaptive immune response against tumor cells and prevent future relapse. The drugs chosen for our combination target metabolic pathways that cancer cells are heavily dependent upon and do not interact with or induce mutations in DNA. These properties place Trifecta at the forefront of developing anticancer therapies. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2020. / FAU Electronic Theses and Dissertations Collection

Osteoarthritis (OA) is a common and incapacitating joint disease beginning with breakdown of articular cartilage and extending into subchondral bone. At present, the processes through which the disease occurs are poorly understood, and interventions are limited to pain relief and eventual joint replacement. OA is commonly associated with obesity and corresponding pathologies, and as OA is demonstrably not a product of passive erosion of cartilage over time or under increased loads there must needs be some other mechanistic link between the two conditions. We hypothesize that the production of ceramides, a hallmark of the insulin resistance syndrome underlying many obesity-related conditions, acts to induce OA through its pro-inflammatory and pro-apoptotic activities, as well as directly inhibiting intracellular mediators of cartilage production and homeostasis. We demonstrate in Wistar rats that a high-fat, high-sugar (HFHS) diet successfully induces OA and that downregulation of ceramide synthesis through intraperitoneal myriocin administration does not prevent this degradation, and that myriocin in conjunction with a standard chow diet actually induces OA. Alteration in OA biomarkers in this study are discussed. We then tested the efficacy of a topical regimen of wogonin, an anti-inflammatory, anti-oxidative, and potentially analgesic compound in a surgical destabilization model (DMM) of OA in mice and demonstrate its disease modifying anti-OA properties. We further test the efficacy of this compound on the HFHS model previously established and find it successfully ameliorated the morphology and biomarker changes associated with OA; based on this data we hypothesize that inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is the most relevant physiological target of wogonin in a HFHS-induced OA model. Lastly and separately, we seek to clarify conflicting data regarding secondhand smoke (SHS), which observational studies suggest having either deleterious or beneficial effects to preexisting OA. In the first controlled study on the subject we model we demonstrate in a murine DMM model that SHS accelerates cartilage degradation and patterns of biomarker expression characteristic of OA, eliminating the question of any potential benefits of SHS to articular cartilage.

Natural bioactive compounds have drawn the interest of many researchers worldwide in their effort to find novel treatments, including prostate cancer (PC) treatment which is estimated to be 13.1% of all new cancer cases in the U.S. in 2021. Many of these bioactive compounds have been identified from treatments in traditional Chinese medicine (TCM), that often have multiple bioactive compounds present. However, in vitro studies frequently focus on the compounds in isolation, or in simple combinations of two compounds. We used mixture design response surface methodology (MDRSM) to assess changes in PC cell viability after 48 hours of treatment to identify the optimal mixture of all 35 three-compound combinations of seven bioactive compounds from TCM. We used Berberine, Wogonin, Shikonin, Curcumin, Triptolide, Emodin, and Silybin to treat PC-3, DU145, and LNCaP human PC cells, and a drug-resistant PC-3 cell line. Berberine and Wogonin most frequently contributed to the optimal combination to reduce cell viability in PC-3 and LNCaP cells; DU145 cells more frequently responded best to a single compound.