Bachelor of Science (BS), Ohio University, 2022, Biological Sciences

Lung cancer is the second most prevalent cancer in both men and women in the United States, and it currently the leading cause of cancer-related deaths. While there are current standard treatments for lung cancer, these treatments are not effective enough, and can lead to drug resistance, severe side effects, and even recurrence of the cancer and death. It is easy to see that better anticancer therapies are urgently needed. It has been shown that many cancer types, including Non-Small Cell Lung Cancer (NSCLC), which makes up 84% of all lung cancer diagnosis, are addicted to glucose and are sensitive to glucose deprivation, ultimately resulting in cancer cell-death. DRB18 is a small molecule compound developed at Ohio University that has been shown to target and inhibit glucose transporters (GLUTs), which are responsible for the uptake of glucose into cells, particularly cancer cells. DRB18 has been shown to be effective in inhibiting cancer cell growth in vitro (cell culture) and in vivo (nude mice) without noticeable side effects. While this may be a promising anticancer therapy by itself, combining DRB18 with an FDA-approved anticancer drugs Paclitaxel, Trametinib, or Brigatinib maximized the efficacy of the treatment in vitro in NSCLC A549 cells, and combination treatment of DRB18 + Paclitaxel resulted in shrunken A549 xenograft tumors in vivo, without increasing unwanted side effects. This combination therapy has the potential to benefit cancer patients for decades to come.

Committee: Soichi Tanda (Advisor); Xiaozhuo Chen (Advisor) Subjects: Biology; Biomedical Research; Medicine; Oncology

Doctor of Philosophy, The Ohio State University, 2022, Biomedical Sciences

Small cell lung cancer (SCLC) is an extremely aggressive neuroendocrine tumor, accounting for approximated 13% of all lung cancer cases. SCLC is characterized by rapid growth and early metastasis. Despite marked improvements in the number and efficacy of targeted, therapeutic options and overall survival rates in SCLC have remained nearly unchanged for almost three decades. The lack of significant progress can be attributed to our poor understanding of the biology of SCLC. Although immune checkpoint inhibitors were recently approved as front-line therapies for SCLC, we still need to better understand the mechanisms responsible for the selective vulnerability of some SCLCs to these inhibitors. Recent work utilizing sequencing data and single cell analyses identified four distinct subsets of SCLC, based on the expression levels of the transcription factors ASCL1, NEUROD1, POU2F3 and YAP1. Each subset was found to have its own distinct biology and therapeutic vulnerabilities. However, these subsets appear to be phenotypically unstable, representing snapshots in the gradual evolution of a tumor that exhibits significant plasticity. Tumor evolution, a product of this plasticity, results in the emergence of significant intratumoral heterogeneity which plays an important role in multiple aspects of SCLC development and progression, including cell survival and proliferation, metastasis and angiogenesis. The recent paradigm shifting discoveries in the biology of SCLC are now beginning to inform the design of new therapeutic strategies for the management of this intractable disease. Receptor Tyrosine Kinase-like Orphan Receptor 1 (ROR1) is an oncofetal protein that is emerging as a therapeutic target and is co-expressed with BCL2 in multiple tumor types due to microRNA coregulation. We hypothesize that ROR1-targeted therapy is effective in small cell lung cancer and synergizes with therapeutic BCL2 inhibition. Tissue microarrays (TMAs) and formalin-fixed paraffin-embedded (F (open full item for complete abstract)

Committee: David Carbone (Advisor); Matthew Ringel (Committee Member); Carlo Croce (Committee Member); Philip Tsichlis (Committee Member) Subjects: Biology; Biomedical Research; Oncology

Doctor of Philosophy, University of Akron, 2017, Chemistry

Imidazolium salts have received significant attention for their anti-cancer properties. Human cancer cell lines have been treated with hundreds of imidazolium salts and many have shown promise for clinical potential. The leader of this class is YM155, a survivin suppressor that has gone through clinical trials but is not yet approved by the Food and Drug Administration (FDA) for the treatment of cancer. Numerous studies have been compiled to create structure activity relationships addressing what functional groups can help increase anti-proliferative effects and which functional groups produce imidazolium salts with weak anti-cancer properties. The general trend throughout the literature is that lipophilicity increases anti-cancer potential. Unfortunately, little is known about the mechanism of action, cellular target, and specificity of imidazolium salts which are all limiting factors towards progression into clinical applications. Chapter II describes the synthesis and characterization of a series of N,N'-bis(naphthylmethyl) imidazolium salts. These compounds were also tested for their in vitro anti-cancer properties against several non-small cell lung cancer cell lines and found to be highly active. Each compound had low aqueous solubility, but could be solubilized by a cyclodextrin that is FDA approved for drug formulations. In vitro mechanism of action studies were also performed on one compound and suggested the compound induced an apoptotic mode of cell death. Chapter III provides the synthesis and characterization of a related series of compounds with higher aqueous solubility. The major findings included the incorporation of quinolylmethyl moieties, similar to naphthylmethyl substituents, to increase aqueous solubility without drastically lowering anti-cancer activity. It was also determined that the anion plays a significant role in the aqueous solubility of these imidazolium salts. Chapter IV presents the synthesis, characterization, and in vitr (open full item for complete abstract)

Committee: Wiley Youngs Dr. (Advisor); Claire Tessier Dr. (Committee Member); Ziegler Chris Dr. (Committee Member); Shriver Leah Dr. (Committee Member); Hossein Tavana (Committee Member) Subjects: Biochemistry; Chemistry; Medicine

Master of Science (MS), Wright State University, 2023, Biochemistry and Molecular Biology

Extracellular signal related kinase 3 (ERK3) is one of the atypical mitogen activated protein kinases (MAPK). It is expressed ubiquitously and plays a role in a variety of cellular processes, including cell growth and differentiation. ERK3's role in promoting migration and invasion in various cancers has been well established. ERK3 is upregulated in non-small cell lung cancers (NSCLCs) and has been shown to promote NSCLC tumor growth and progression. However, the regulation of ERK3 in lung cancers remains largely unclear. A recent study indicates that ERK3 phosphorylation at S189, an indicator of ERK3 activity, is upregulated by KRAS in NSCLCs. KRAS is one of the most commonly mutated oncogenes in lung cancers. To study the KRAS dependent regulation of ERK3, knockdown of KRAS was performed and it resulted in a remarkable reduction in ERK3 phosphorylation as well as total ERK3 protein level confirming the regulation of ERK3 by KRAS. Upon knockdown of KRAS a significant reduction of ERK3 mRNA level was observed indicating that KRAS regulates ERK3 at transcriptional level. Further, we found that the regulation of ERK3 by KRAS may be through the transcription factor c-Jun, that is well-known to be activated by KRAS signalling. Our data indicates that c-Jun positively regulates ERK3 transcription in LUAD cell lines. Further, we have found that KRAS upregulates c-Jun activating phosphorylations in LUAD cells, suggesting that KRAS regulates ERK3 through c-Jun. Given the discrepancy regarding the role of ERK3 in NSCLC cell growth reported in previous studies, we have thoroughly investigated the role of ERK3 in cell growth by stable knockdown of ERK3 using shRNA targeting different regions of ERK3 mRNA, as well as by using ERK3 inhibitors in a variety of NSCLS cell lines. While knockdown of ERK3 via targeting the coding region did not affect cell proliferation, targeting the 3'UTR of ERK3 or treatment with ERK3 inhibitors reduced the proliferation of LUAD cells.

Committee: Weiwen Long Ph.D. (Advisor); Kwang-Jin Cho Ph.D. (Committee Member); Michael Craig Ph.D. (Committee Member) Subjects: Biochemistry; Molecular Biology

Doctor of Philosophy, The Ohio State University, 2021, Biomedical Engineering

Lung cancer is leading cause of cancer-related deaths in the United States with 5-year survival rate of 18.6%. This is due to late detection of lung cancer and problems in screening for lung cancer. Indeterminate pulmonary nodules (IPNs) are pulmonary nodules size between 7-20mm diameter solid nodules. 90% of IPNs are incidentally found and they are hard to diagnosis due to their small size and current diagnosis methods such as CT, PET scans and biopsy involve high exposure to radiation or invasive and could lead to complications. The majority of lung cancer patients have non-small cell lung cancer (NSCLC) and 64% of these patients exhibit driver mutations such as epithelial growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK) and Ras mutations. These patients have shown to have improved survival rate if they are treated with targeted therapies directed against the driver mutations. Although these patients initially show strong response to targeted therapies, most patients develop resistance to these targeted treatments through secondary point mutation and epithelial to mesenchymal transition (EMT). The lung is a dynamic organ where alveolar epithelial cells are normally exposed to significant mechanical forces (i.e. ~8% cyclic strain, transmural pressure and shear stress) while primary lung tumor cells experience a 40-fold decrease in these mechanical forces/strain. Although biomechanical factors in the tumor microenvironment have been shown to be a significant driver of cancer progression, there is limited information about how biophysical forces alters drug sensitivity in lung adenocarcinoma cells. Based on the known importance of mechanical forces/strain on lung injury and repair and the significant difference in cyclic strain applied to normal and cancer cells in the lung, we hypothesized that cyclic mechanical strain would activate important oncogenic pathways and alter drug sensitivity. Although local mechanical properties of the lung tumor may (open full item for complete abstract)

Committee: Ghadiali Samir Dr. (Advisor); Joshua Englert Dr. (Committee Member); Arunark Kolipaka Dr. (Committee Member) Subjects: Biomedical Engineering

Doctor of Philosophy, The Ohio State University, 2019, Biomedical Sciences

LKB1 is the fourth-most frequently mutated gene in lung adenocarcinoma, with loss-of-function observed in up to 30% of all cases (Collisson et al. 2014; Kaufman et al. 2014). Our previous work identified a 16-gene signature for LKB1 loss of function through not just mutation, but deletion and epigenetic silencing, which occurs relatively frequently (Lee et al. 2013; Kaufman et al. 2014). We applied this genetic signature to lung adenocarcinoma samples in the Cancer Genome Atlas (TCGA) and discovered a novel association between LKB1 loss and widespread CpG demethylation. LKB1-loss tumors also expressed significantly less DNA methyl transferase (DNMT1) and show depletion of S-Adenosyl-Methionine (SAM-e), which is the primary substrate for DNMT1 activity. Repetitive element transcriptional start sites are demethylated and sensitivity to azacytidine is lower in LKB1 loss. Mechanistically, demethylated CpGs are enriched for FOXA1/2/3 consensus binding sites, and we further identified that FOXA localization and turnover is dependent upon LKB1 and the downstream kinase SIK. Overall, these findings demonstrate that a large number of lung adenocarcinoma patients have a unique epigenetic profile driven by LKB1 loss which could play a role in lung tumorigenicity and resistance to immunotherapy.

Committee: David Carbone (Advisor); Christopher Oakes (Committee Member); Matthew Ringel (Committee Member); Susan Cole (Committee Member); Sameek Roychowdhury (Committee Member) Subjects: Bioinformatics; Biology; Biomedical Research; Genetics; Oncology

Doctor of Philosophy, Case Western Reserve University, 2018, Molecular Medicine

The sustainable activation of the RAS/MAPK and PI3K/AKT signaling pathways in cancer is promoted by a reduction in the activity of the tumor suppressor protein phosphatase 2A (PP2A). Therefore, a novel therapeutic strategy consists of directly activating PP2A, leading to the simultaneous inhibition of these oncogenic pathways. Our lab has successfully developed first-in-class Small Molecule Activators of PP2A (SMAPs), which induce tumor growth inhibition in vivo. Alterations to the putative drug binding site validate PP2A as the direct target of SMAPs. The putative residues of PP2A-Aa that were interacting with SMAPs K194 E197, and L198 were mutated. H358, a KRAS-driven lung adenocarcinoma cell line, was used to create isogenic cell lines stably overexpressing mutated and wild type PP2A-Aa. SMAP response was investigated in vivo using a xenograft model of H358 isogenic cell lines and it was determined that tumors harboring mutant K194R and L198V PP2A-Aa were resistant to SMAPs treatment. Together, our results suggest that residues K194 and L198 are required for drug binding and subsequent target engagement. On another hand, most lung adenocarcinoma (LUAD) patients acquire resistance to tyrosine kinase inhibitors (TKI) via mechanisms enabling the sustained activation of the MAPK and PI3K oncogenic pathways downstream of the tyrosine kinase EGFR. We hypothesize that activation of PP2A simultaneously inhibits the MAPK and AKT pathways and is a promising therapeutic strategy for TKI-resistant LUAD. TKI-resistant LUAD cell lines were treated with SMAPs. RNAseq kinase enrichment analysis followed by principal component analysis indicated that SMAP treatment induces a gene signature similar to a combination of the selective AKT and MEK inhibitors MK2206 and AZD6244, respectively. The therapeutic potential of PP2A activation in vivo was first evaluated in a transgenic mouse model. SMAP- treated mice showed less diffuse lung cancer and a significant decrease in (open full item for complete abstract)

Committee: Goutham Narla (Advisor) Subjects: Biology; Biomedical Research; Cellular Biology; Molecular Biology

Doctor of Philosophy, University of Akron, 2016, Chemistry

Imidazolium salts are unique for their biological activity and application in the field of cancer research. Pharmaceutical research of imidazole-based antitumor compounds has led to the identification of highly effective N,N'-substituted imidazolium salts with comparable activity to that of cisplatin, which has been used as a chemotherapeutic agent for decades. The high toxicity of present cancer treatments has prompted the investigation into other compounds with similar efficacy. Select N,N'-bis(arylmethyl)-imidazolium salts have been shown to exhibit high anti-proliferative activity, yet these compounds have inferior aqueous solubilities. This dissertation describes the rational design and biological activity of N,N'-bis(arylmethyl)imidazolium salts with hydrophilic and hydrophobic substituents on the imidazole ring for the treatment of lung carcinomas and chronic urinary tract infections. Chapter I discusses the background and development of N,N'-substituted imidazolium salts within the field of antitumor research. Chapter II addresses the synthesis and anti-proliferative activity of C4(C5) substituted N,N'-bis(arylmethyl)-imidazolium salts with hydrophilic or lipophilic groups on the imidazole ring. In vitro activity of these seventeen imidazolium salts against select non-small cell lung cancer cell (NSCLC) lines (NCI-H460, HCC827, and NCI-H1975) was analyzed by biological assays. It was determined that the phenyl substituents at the C4(C5) positions increased the in vitro antitumor activity of these salts to create highly active compounds. Chapter III is designated to the synthesis and anti-proliferative activity of thirty-one N,N'-bis(arylmethyl)benzimidazolium salts against NCI-H460, HCC827, NCI-H1975, and NCI-A549 lung cancer cell lines. Our data confirms that naphthylmethyl substituents at the nitrogen atoms (N1(N3)) and highly lipophilic substituents at the carbon atoms (C2 and C5(C6)) can generate benzimidazolium salts with anti-proliferative activity c (open full item for complete abstract)

Committee: Wiley Youngs Dr. (Advisor); Claire Tessier Dr. (Committee Member); Chrys Wesdemiotis Dr. (Committee Member); David Modarelli Dr. (Committee Member); Yang Yun Dr. (Committee Member) Subjects: Chemistry

Doctor of Philosophy in Medical Sciences (Ph.D.), University of Toledo, 2005, College of Graduate Studies

Targretin was tested for its ability to prevent lung tumors in A/J mice induced by vinyl carbamate. Doses 30-300 mg/kg decreased tumors multiplicity by greater than 19%. Administering 200 mg/kg Targretin from weeks 4-19 and 4-25 decreased the multiplicity of tumors from 35.3 ± 1.43 to 29.1 ± 1.51 and 25.0 ± 0.93, respectively. Targretin decreased tumor size in 4-25 and 23-25 wk prevented and reversed DNA hypomethylation in lung tumors. Microarrays analysis indicated that Targretin short-term treatment had a greater effect on mRNA expression than long-term treatment. In lung tumors, Targretin decreased expression of p16INK4, EP3, Caspase-3, Dnmt-3, survivin, Cyclin B2, Cyclin E1, iNOS and ER-"Α", with short-term treatment being more effective than long-term treatment. Short-term treatment increased expression of Apolipoprotein D, Cyp26b, and Fabp-4, while long-term treatment only increased expression of Apolipoprotein D and Cyp26b. Budesonide and R115777 were evaluated for their usefulness in preventing vinyl carbamate-induced lung tumors in A/J mice. One week after the second dose of vinyl carbamate, mice received 60 or 100 mg/kg R115777 (gavage), 0.8 or 1.6 mg/kg budesonide (diet), or the two drugs combined treatment until sacrificed at 20, 28 and 36 weeks. Other mice received the drugs for two weeks prior to sacrifice at 20 wk. The rank order for prevention of lung tumors was the combined treatment > budesonide > R115777 (only at 20 wk). The drugs prevented DNA hypomethylation in lung tumors. While p21waf1/cip1 and Hoxa5 were methylated, APC, EGFR, RAR-"Β", DAPK and Fhit genes were not.

Committee: Michael Pereira, Ph.D. (Advisor) Subjects:

PHD, Kent State University, 2024, College of Arts and Sciences / Department of Biological Sciences

Dendritic cell (DC)-based anti-cancer vaccines have been ineffectual as a monotherapy for invasive cancer. We therefore formulated a combined therapy approach that exploits the impeccable T cell-sensitizing capacity of dendritic cells and either of two small molecule inhibitor drugs, with the expectation of improved anti-cancer activity. The first, Lapatinib, is a potent inhibitor of epidermal growth factor receptor-family kinases. In conjunction with in vitro vaccine surrogates interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α), Lapatinib synergistically enhanced metabolic suppression for a panel of murine breast carcinoma lines. In addition, this combination increased cell death significantly over single treatments, with the presence of several cellular markers (Annexin V, Propidium iodide staining and executioner caspase activation) consistent with an apoptotic mechanism. Additionally, in an orthotopic model of HER-2 breast cancer, combination therapy resulted in significant tumor regression compared to single treatments, including a trend towards extended survival. Assessment of specific tumor infiltrating lymphocyte (TIL) subpopulations within tumors showed insignificant infiltrates into tumors from either untreated or Lapatinib-only treated mice, substantial and expected increases in various lymphocyte populations at both tumor margins and infiltrating into the tumor in mice that received DC-based vaccination alone and also in mice receiving both DC vaccination and Lapatinib. This finding implicates that Lapatinib is not immunomodulatory by itself but suggests that it may sensitizes tumor cells to the lymphocyte-produced cytokines already being secreted at the site of disease. The second tested drug was the novel imipridone, TIC10/ONC201. It was evaluated in conjunction with Th1 cytokines in a panel of phenotypically diverse human breast and human lung cancer cell lines. When combined with the Th1 cytokine effectors, TNF-α and IFN-γ, TIC10/ONC201 (open full item for complete abstract)

Committee: Gary Koski (Committee Chair); Manabu Kurokawa (Committee Member); Yaorong Zheng (Committee Member); Soumitra Basu (Committee Member); Colleen Novak (Committee Member) Subjects: Biology; Biomedical Research; Immunology; Molecular Biology; Oncology

Bachelor of Sciences, Ohio University, 2023, Biological Sciences

Globally, metastasis causes approximately 90% of mortality in cancer, making it a leading cause of death. In the United States, in both men and women, lung cancer is the second most prevalent cancer, with over 283,000 new cases estimated to be diagnosed in 2023. Both the tumor microenvironment (TME) and macropinocytosis have been shown to play a role in invasion, proliferation, and recurrence of cancers. Dr. Xiaozhuo Chen's lab at Ohio University studied the effects of the TME on cancer cells by performing RNA sequencing on A549. A549 are non-small cell human lung cancer (NSCLC) cells, which showed a consistent, significant upregulation of Stanniocalcin 1(STC1) gene expression when treated with extracellular ATP (eATP) and TGF-β. STC1 is a protein hormone involved in the regulation of the calcium phosphate balance, as well as ATP synthesis in mitochondria within the cell. Further studies showed that knock down of the STC1 gene led to reduced invasion and proliferation when compared to the untreated A549 cells. The aim of this project was to perform two main studies; one, to identify and assess macropinocytosis in a variety of cancer cell lines, and two, to investigate the effects of the STC1 gene on macropinocytosis. Using ATP concentration assays and IPA3 inhibition assays, macropinocytosis was examined in 11 cancer cell lines of varying cancer types. Macropinocytosis was confirmed with fluorescence microscopy by the colocalization of green fluorescent ATP and red fluorescent dextran. The impact of the knock-out of STC1 on macropinocytosis in A549 cells was investigated and quantified using ImageJ. The fluorescence microscopy study revealed that STC1 gene did play a role in macropinocytosis as predicted, which may be important for its effect on proliferation and invasion, the first step of metastasis.

Committee: Xiaozhuo Chen PhD (Advisor); Janet Duerr PhD (Advisor) Subjects: Biology; Cellular Biology; Molecular Biology

Doctor of Philosophy (PhD), Wright State University, 2023, Biomedical Sciences PhD

Ras proteins are small GTPases that regulate cell proliferation, differentiation and survival at the plasma membrane (PM). There are three Ras isoforms ubiquitously expressed in mammalian cells: H-, N- and K-Ras. Constitutively active Ras mutations are found in ~19% of all human cancers, with ~75% of those being in K-Ras. There are K-Ras inhibitors in clinic but they only target the oncogenic K-RasG12C mutant, which only makes up a small sub-set of K-Ras-driven cancers. Thus, there still exists a need for a pan anti-K-Ras drug. Ferrocene derivatives are a class of compounds that have been shown to inhibit the growth of a lung cancer cell line harboring an oncogenic mutant K-Ras via their elevation of cellular reactive oxygen species (ROS). Given that lung cancer cells harboring an oncogenic mutant K-Ras require K-Ras signaling for their survival, the effect of a novel ferrocene derivative, C16H20FeClNO, on K-Ras signaling was tested. C16H20FeClNO was found to specifically disrupt the K-Ras/MAPK signaling pathway and inhibit the growth of K-Ras-dependent human pancreatic and lung cancer cell lines. Given that localization to the PM is essential for K-Ras signaling, the effect of C16H20FeClNO on K-Ras PM localization was tested. C16H20FeClNO was found to dissociate K-Ras, but not other Ras isoforms, from the PM. Furthermore, other ROS-elevating drugs had a similar effect on K-Ras PM localization and K-Ras signal output. C16H20FeClNO was shown to elevate cellular ROS levels and supplementation of N-acetylcysteine (NAC), a general antioxidant, reversed these effects, suggesting that C16H20FeClNO disrupts K-Ras PM localization and K-Ras signaling through an ROS-mediated mechanism. Lastly, K-Ras histidine 95 (His95) residue in the G-domain was identified as the oxidative residue involved in C16H20FeClNO-induced K-Ras dissociation from the PM. Overall, C16H20FeClNO disrupts K-Ras PM localization and signal output through oxidation of His95. Taken together, this dissertatio (open full item for complete abstract)

Committee: Kwang-jin Cho Ph.D. (Advisor); David R. Ladle Ph.D. (Committee Member); Kuppuswamy Arumugam Ph.D. (Committee Member); David Cool Ph.D. (Committee Member); Michael Leffak Ph.D. (Committee Member) Subjects: Biomedical Research

Doctor of Philosophy (PhD), Ohio University, 2020, Biological Sciences (Arts and Sciences)

Deregulated uptake of extracellular nutrients such as glucose has been named as a hallmark of cancer metabolism in recent years. Cancer cells drastically increase the uptake of glucose by overexpressing membrane-spanning glucose transporter proteins (GLUTs) to meet their energy and biomass synthesis demands and are very sensitive to glucose deprivation. Therefore, targeting glucose uptake via GLUTs has become an attractive anticancer strategy. We previously reported a small molecule, WZB117, that inhibits GLUT1-mediated glucose uptake in A549 lung cancer cells in vitro and in vivo. To correct the problem of chemical instability of WZB117, here we report a much more stable WZB117-derived inhibitor, DRB18, synthesized after another round of structure activity relationship study. In this study we tested the hypothesis that the more stable DRB18 reduces cell growth in human cancer cells in vitro and in vivo by targeting GLUT1-4. We performed cell viability and glucose uptake assays in different cancer cell lines treated by DRB18. We further tested the GLUT1-4 inhibitory potency of DRB18 by performing glucose uptake in GLUT-specific cell lines. Docking analysis was used to determine the binding energies and interacting residues of DRB18 and hGLUT1-4. Western blot was used to investigate how DRB18 affects expression of hGLUT1-4 in vitro. Metabolomics analysis was performed to detect the qualitative and quantitative effects of DRB18 on metabolic pathways in cancer cells. Cancer cell xenograft tumor study was done in nude mice to demonstrate the anticancer efficacy of DRB18 in vivo. This was followed by metabolomics analysis in vivo and use of immunohistochemistry (IHC) to understand the effect of DRB18 on GLUT1-4 expression in vivo. CRISPR-Cas9 knockout of the GLUT1 gene was used to determine its role in glucose uptake and cellgrowth in vitro and in vivo. Statistical analysis was performed using Student's t-test and one-way ANOVA. Here we report that, DRB18 reduced gluco (open full item for complete abstract)

Committee: Xiaozhuo Chen Dr. (Advisor); Stephen Bergmeier Dr. (Committee Chair); Robert Colvin Dr. (Committee Member); Shiyong Wu Dr. (Committee Member) Subjects: Biology; Molecular Biology

Bachelor of Science (BS), Ohio University, 2020, Biological Sciences

ATP in the tumor microenvironment is 1,000-10,000 times higher than in normal tissues and has recently been found to play many roles in tumorigenesis, including metastasis, which is associated with up to 90% of cancer related death. Metastasis starts with a process called epithelial to mesenchymal transition (EMT). Our lab has previously described that internalization of extracellular ATP (eATP) by cancer cells (in vitro and in vivo) by macropinocytosis in human cancer cells elevates intracellular ATP levels, and enhances cell proliferation and resistance to anticancer drugs. Most recently, we have reported ATP's roles in inducing EMT and other early metastatic activities independent of TGF-beta in human lung cancer cells. Traditionally, TGF-beta has been considered to be important and necessary for EMT induction, but our published and more recent preliminary findings have prompted us to reconsider ATP as an alternative master inducer and regulator of EMT. In this study, we tested the hypothesis that ATP, like TGF-beta, is a master inducer and regulator of EMT, triggering EMT in a fashion similar to but non-identical to TGF-beta. Transwell assays revealed that addition of extracellular ATP (eATP) to human lung cancer cells induced both cell migration and invasion at higher rates than those of TGF-beta. Fluorescence microscopy exhibited that eATP treatment induced cell morphology changes indicative of EMT. ATP assays showed that eATP treatment resulted in significantly elevated intracellular ATP levels while TGF-beta treatment did not change the iATP level. This result indicates that although the treatments of eATP or TGF-beta all result in migration and invasion, they also lead to some different intracellular responses. The combined cell treatment with lower concentrations of eATP and TGF-beta led to some noticeable additive effect in invasion activity, suggesting that these two molecules may induce invasion by activating EMT through similar but non-identical sig (open full item for complete abstract)

Committee: Xiaozhuo Chen Dr. (Advisor) Subjects: Biology; Biomedical Research; Cellular Biology; Molecular Biology; Oncology

Doctor of Philosophy, Case Western Reserve University, 2020, Biomedical Engineering

As the arsenal of therapeutic strategies in the fight against cancer grows, so too does the need for predictive biomarkers that can precisely guide their use in order to match patients with their optimal personalized treatment plan. Currently, clinicians often have little recourse but to initiate treatment and monitor a tumor for signs of response or progression, which exposes non-responsive patients to overtreatment, harmful side effects, and windows of ineffective therapy that increase a patient's risk of progression or metastasis. Thus, there is an urgent need for new sources of predictive biomarkers to help more effectively plan personalized treatment strategies. Radiological images acquired before treatment may contain previously untapped predictive information that can be quantified in the form of computational imaging biomarkers. The vast majority of existing computational imaging biomarkers provides analysis limited to the tumor region itself. However, the tumor environment contains critical biological information pertinent to tumor progression and treatment outcome, such as tumor-associated vascularization and immune response. This dissertation focuses on the development of new, biologically-inspired computational imaging biomarkers targeting the tumor environment for the prediction of response to a wide range of chemotherapeutic and targeted treatment strategies in oncology. First, we explore measurements of textural heterogeneity within the tumor and surrounding peritumoral environment, and demonstrate the ability to predict therapeutic response and tumor biology to neoadjuvant chemotherapy in primary and targeted therapy in primary and metastatic breast cancer. Second, we introduce morphologic techniques for the quantification of the twistedness and organization of the tumor-associated vasculature, and demonstrate their association with response and survival following four different therapeutic strategies in breast cancer MRI and non-small cell lung canc (open full item for complete abstract)

Committee: Madabhushi Anant (Advisor); Wilson David (Committee Chair); Abraham Jame (Committee Member); Gilmore Hannah (Committee Member); Plecha Donna (Committee Member); Varadan Vinay (Committee Member) Subjects: Biomedical Engineering; Biomedical Research; Computer Science; Medical Imaging; Medicine; Oncology; Radiology

Doctor of Philosophy, Case Western Reserve University, 2020, Pharmacology

A governing principle of cancer development is defined by a coordinate gain of oncogenic function and loss of tumor suppressor activity. To fully reverse this pathogenic process, one would want to simultaneously inhibit oncogene activity while reengaging tumor suppressor function. However, the majority of targeted therapies are directed at modulating the oncogenic gain with few therapies directed at the critical tumor suppressor proteins. This is based upon the dogma that, in a cell, it is easier to turn something off than to turn something back on. Indeed, activation of tumor suppressors using pharmaceutically tractable approaches have proven to be challenging. Yet, efforts persist to develop activators of tumor suppressor proteins. One that stands out as a therapeutic target is Protein Phosphatase 2A (PP2A). PP2A is a serine/threonine phosphatase involved in the regulation of many cellular processes and is genetically altered or functionally inactivated in many cancers highlighting its central role in cancer pathogenesis. One of the best-defined substrates of PP2A is the transcription factor c-MYC (MYC). MYC, a well-described oncogene, is activated through both genetic amplification and stabilizing post-translational modifications. Cancers associated with high MYC expression are generally more aggressive. However, MYC has remained an elusive drug target as it lacks targetable drug pockets. MYC is rapidly degraded and its activity is inhibited by active PP2A. Thus, PP2A reactivation is a proposed strategy for the treatment of MYC driven cancers. Small Molecule Activators of PP2A (SMAPs) have been recently described for their potent anti-cancer activity which is dependent upon their ability to activate PP2A, reengaging its tumor suppressor activity. This research demonstrates that activation of PP2A by SMAPs leads to MYC degradation resulting in the inhibition of cancer growth in both cellular and in vivo model systems. Biochemical and genetic tools ar (open full item for complete abstract)

Committee: Goutham Narla M.D./Ph.D (Advisor); Ruth Keri Ph.D (Committee Chair); Marvin Nieman Ph.D (Committee Member); Amar Desai Ph.D (Committee Member); David Wald M.D. /Ph.D (Committee Member) Subjects: Biomedical Research; Pharmacology

Master of Science (MS), Wright State University, 2019, Biochemistry and Molecular Biology

Mitogen activated protein kinases (MAPKs) are Ser/Thr kinases that relay the extracellular signal into intracellular responses and regulate several biological responses. They are classified into conventional MAPKs and atypical MAPKs. Extracellular signal regulated kinase 3 (ERK3) is an atypical MAPK that has a single phospho-acceptor site (Ser 189) in its activation motif instead of the canonical Thr-Xaa-Tyr (TXY) motif of conventional MAPK like ERK1/2. ERK3 comprises of a unique C terminal tail and a central C34 domain that further distinguishes it from ERK1/2. Moreover, compared to ERK1/2, much less is known about the upstream activators and the downstream targets of ERK3. Here, our study identifies IL-6 signaling to be negatively regulated by ERK3. We show that ERK3 downregulates the IL-6 target genes and STAT3 phosphorylation. IL-6 is a multifunctional pleiotropic cytokine that signals predominantly via JAK/STAT signaling pathway. In response to IL-6, STAT3 is phosphorylated at Tyrosine 705 (Y705) residue by JAK2, which successively activates STAT3 as a transcription factor. The IL-6/STAT3 pathway is negatively regulated by a feedback inhibitor SOCS3 (Suppressor of Cytokine Signaling 3) which binds to JAK2, hence, preventing the phosphorylation and activation of STAT3. Interestingly, this study reveals that ERK3 interacts with SOCS3 via its C34 domain. Until now, there has been no information about the physiological or the biochemical function of the C34 domain of ERK3. Hence this study has identified a novel role of ERK3's C34 domain. Furthermore, we show that ERK3 facilitates the interaction between SOCS3 and JAK2, promoting the inhibition of STAT3 Y705 phosphorylation and activation. Taken together, the findings of our study provide important and novel insights into the negative regulation of IL-6/STAT3 signaling pathway by ERK3.

Committee: Weiwen Long Ph.D. (Advisor); Madhavi Kadakia Ph.D. (Committee Member); Hongmei Ren Ph.D. (Committee Member) Subjects: Biochemistry; Biology; Biomedical Research

Doctor of Philosophy (PhD), University of Toledo, 2019, Biomedical Sciences (Medical Microbiology and Immunology)

Cancer patients are known to be at high risk for thrombosis, a leading cause of death among cancer patients. Cancer patients are also at increased risk of developing recurrent thrombosis or severe bleeding as a result of anticoagulation therapy to treat cancer-associated thrombosis. Platelet-leukocyte aggregates (PLAs) are associated with increased thrombosis risk, but it remains unclear how PLA formation may influence thrombosis in the context of cancer. We tested the hypothesis that lung cancer patient platelets differ from healthy volunteers. Increased platelet counts can be a significant predictor of history of thrombosis in lung cancer patients. Lung cancer patient platelets expressed more P-selectin, a marker of activation, than healthy volunteers did (mean fluorescence intensity of P-selectin 29.12 and 11.69 units, respectively). We compared PLA formation in lung cancer patients compared to healthy volunteers. Lung cancer patients had significantly more CD4+ platelet-T cell aggregates (PTCAs) than healthy controls (36.84% of total CD4+ T cells vs. 19.01%, respectively). The detection limit of >22.43% CD4+ PTCAs was 76.92% sensitive and 72.55% specific for lung cancer. Lung cancer patients had significantly more CD8+ PTCAs than healthy controls (48.41% of total CD8+ T cells vs. 28.77%, respectively), and the detection limit of >35.24% CD8+ PTCAs was 75.00% sensitive and 73.08% specific for lung cancer. T cell-bound platelets were significantly more activated in lung cancer patients compared to healthy controls. Platelets in CD4+ PTCAs expressed MFI of 113.7U in lung cancer patients and 64.67U in healthy volunteers, and the detection limit of >55.15U was 62.22% sensitive and 69.57% specific for lung cancer. Platelets in CD8+ PTCAs expressed MFI of 159.7U in lung cancer patients and 90.35U in healthy volunteers, and the detection limit of >79.95U was 63.04% sensitive and 64.44% specific for lung cancer. We further explored the role of past history of thro (open full item for complete abstract)

Committee: Randall Worth PhD (Committee Chair); R. Mark Wooten PhD (Committee Chair); James Willey MD (Committee Member); Kathryn Eisenmann PhD (Committee Member); Saurabh Chattopadhyay PhD (Committee Member) Subjects: Biomedical Research; Cellular Biology; Immunology; Medicine; Oncology

Doctor of Philosophy, Case Western Reserve University, 2019, Clinical Research

Brain metastases (BM) are the most frequent type of brain tumor, and cause significant morbidity and mortality. As imaging modalities improve, treatments improve and diagnoses increase in frequency, epidemiological studies of BM and its clinical outcomes become increasingly relevant and potentially beneficial to cancer patients. Here, we use large population-level data from the Surveillance, Epidemiology, and End-Results (SEER) program combined with Medicare claims to identify measures of BM incidence, claims algorithm concordance with SEER data, treatment patterns, and treatment efficacy for patients with BM arising from primary lung, breast, or skin cancers. We make use of both traditional epidemiological approaches to minimize confounding, such as age adjustment to a standard population, and more advanced methods, such as propensity score matching. By examining the Medicare claims of patients diagnosed with BM, the following chapters shed light on which patients develop BM, how BM patients benefit from an antineoplastic drug that may simultaneously relieve intracranial hypertension, and which patient populations are underserved with respect to BM treatment.

Committee: Jill Barnholtz-Sloan PhD (Advisor); Fredrick Schumacher PhD (Committee Chair); Jeremy Bordeaux MD (Committee Member); Andrew Sloan MD (Committee Member) Subjects: Biostatistics; Epidemiology; Health Sciences; Medicine; Neurology; Oncology

Doctor of Philosophy, University of Akron, 2018, Chemistry

Imidazolium salts are a group of compounds that have received attention in literature for their ability to inhibit the growth of cancer cells. However, there is a limited amount of information known about how imidazolium salts actually go about killing cancer cells. As more knowledge is gained in this area, more effective imidazolium salt derivatives can be made. Chapter I provides a literature review of N-heterocyclic carbene complexes and the importance of their precursor ligand imidazolium salts in regard to anti-cancer and antimicrobial activity. Contained in Chapter II is a study of lead imidazolium salt compounds and looks at the mode of cell death induced in cancer cells. The Annexin V assay has been utilized to distinguish between the process of apoptosis and necrosis that is caused by the lead imidazolium salts. Results show that the majority of imidazolium salts promote an apoptotic mode of cell death, although this is dependent upon substituents on the imidazole core. Apoptosis was further confirmed using the JC-1 assay to evaluate depolarization of the mitochondrial membrane potential. Furthermore, DNA has been excluded as the potential cellular target of imidazolium salts after studies of potential interactions with calf thymus DNA. Finally, the use of excipients such as 2-hydroxypropyl-β-cyclodextrin to solubilize lipophilic compounds is explored. Chapter III presents the in vivo toxicity evaluation of select imidazolium salts. Results show that two of the tested compounds were received well. Weight recovery was apparent and animals were able to receive multiple injections at 20 mg/kg. The other compounds tested had varying degrees of success, with most animals dying before the study could be completed. Chapter IV provides preliminary data of a new class of imidazolium salts that contain triphenyphosphonium groups. Newly synthesized compounds were determined to have anti-cancer potential. Chapter V describes the in vitro evaluat (open full item for complete abstract)

Committee: Wiley Youngs (Advisor); Claire Tessier (Committee Member); Sailaja Paruchuri (Committee Member); Leah Shriver (Committee Member); Chrys Wesdemiotis (Committee Member) Subjects: Biochemistry; Chemistry