Doctor of Philosophy, The Ohio State University, 2024, Comparative Biomedical Sciences

Over the past 2 decades, the adaptor protein transducin beta-like 1 (TBL1X) has been shown to be upregulated in solid tumors and hematologic malignancies and its overexpression associated with poor clinical outcomes. Moreover, TBL1X dysregulation has been implicated as a key component of oncogenic prosurvival signaling, cancer progression, and metastasis. In addition to its role in regulating major transcriptional programs (e.g. the SMRT/NCOR/BCL6 co-repressor complex, Wnt/β catenin, and NF-κB signaling), the Alinari laboratory has shown that, in diffuse large B-cell lymphoma (DLBCL), TBL1X stabilizes key oncogenic proteins, such as PLK1 and c-MYC, through interaction with a SKP1-CUL1-F-box protein supercomplex. Previously, we showed that targeting TBL1X genetically and pharmacologically with tegavivint (Iterion), a first-in-class small molecule, results in significant DLBCL cell death in vitro and in vivo. Despite this promising evidence of therapeutic potential, cure is not appreciated. As DLBCL is characterized by poor prognosis in the relapsed and refractory (R/R) setting, novel therapeutic options are urgently needed. Given the central role of TBL1X in modulating multiple oncogenic pathways in cancer, we hypothesized that treatment with tegavivint triggers pro-survival signaling, and that combined targeting of compensatory pathways with tegavivint will maximize the therapeutic potential of this agent in DLBCL. Thus, the first section of this dissertation is aimed at identification and mechanistic characterization of synthetic lethal partners that cooperate with tegavivint in DLBCL. Next, given that mantle cell lymphoma (MCL) is an area expertise for our laboratory and the function of TBL1X in MCL was entirely unexplored, we aimed to extend our knowledge of TBL1X as an oncoprotein and therapeutic target into this aggressive and incurable B-cell lymphoma. MCL patients who progress on targeted therapies, such as ibrutinib, have a short survival, and there is thus (open full item for complete abstract)

Committee: Lapo Alinari (Advisor); Robert Baiocchi (Committee Member); Kara Corps (Committee Member); Rosa Lapalombella (Committee Member) Subjects: Cellular Biology; Molecular Biology; Oncology; Pharmaceuticals

Doctor of Philosophy, Case Western Reserve University, 2023, Pathology

Advancements in therapy have improved remission and prolonged survival for patients with B cell malignancies. Unfortunately, for many challenging cancer subtypes such as mantle cell lymphoma, most patients will still relapse and die from the cancer. Chimeric antigen receptor (CAR)-T cell therapy uses genetically modified T cells expressing CAR protein to recognize and kill cancers expressing a specific antigen, such as CD19. Although CD19 CAR-T therapy has been very effective against relapsed/refractory B cell cancers, antigen escape and relapse still occur in up to 40% of patients treated with CD19 CAR-T. This work describes the creation and validation of a novel, BAFF ligand-based CAR that aims to prevent antigen escape by being able to bind multiple BAFF receptors expressed by the cancer cell, rather than one antigen alone. Using cytotoxicity assays, we demonstrate the functionality and specificity of BAFF CAR-T cells in killing several types of B cell malignancies, and we further confirm their efficacy in several mouse models of cancer. These results have helped bring BAFF CAR-T therapy to Phase I clinical trials, and we hope they prove efficacious for patients with relapsed/refractory B cell malignancies. Meanwhile, interest has rapidly increased in natural killer (NK) cell-based immunotherapies. NK cells have important roles in cancer immunosurveillance and offer advantages over T cells in clinical safety and time-to-treatment. However, NK therapies face challenges in efficacy and persistence, often due to the presence of TGF-β, a powerful immunosuppressive cytokine that is frequently elevated in cancer patients and a primary cause of NK cell dysfunction. In this work, we characterize an undiscovered role of cyclin dependent kinase 5 (Cdk5) and its coactivator p35 in NK cell cytotoxicity. Using genetic tools to modulate Cdk5/p35 kinase activity in human NK cells, as well as using NK cells from p35 knockout mice, we show that Cdk5/p35 negatively regulates NK (open full item for complete abstract)

Committee: Reshmi Parameswaran (Advisor); Alex Huang (Committee Chair); John Letterio (Committee Member); Clive Hamlin (Committee Member) Subjects: Biology; Biomedical Research; Immunology; Medicine; Molecular Biology; Oncology

Doctor of Philosophy, The Ohio State University, 2023, Biomedical Sciences

Mantle Cell Lymphoma (MCL) is a B cell lymphoma distinguished from other B cell lymphomas by a t(11;14)(q13:q32) translocation juxtaposing the CCND1 gene downstream of the immunoglobulin heavy locus (IgH) promoter. CCND1 and SOX11, a neural transcription factor, are the most commonly upregulated genes in MCL. The average patient with MCL is ~70 years of age and male with a 3:1 skew in sex. Current treatments include induction therapy with chemotherapeutics, first-line therapy with targeted agents such as ibrutinib, and additional lines with more experimental agents. Stem cell transplant is possible after induction therapy in a small percentage of patients due to the physical strain. Relapse is basically universal with targeted therapies and frequently occurs after stem cell transplant. MCL is considered incurable today. Many clinical trials are ongoing to develop novel treatment strategies. Many of the targeted therapy trials utilize the BCL-2 inhibitor venetoclax, an FDA-approved agent for use in CLL, and second or third-generation BTK inhibitors such as acalabrutinib. Immunotherapy is a rapidly expanding field for MCL, with many trials using CD19 CAR T cells, bi- or tri-specific antibodies, and drug-antibody conjugates. Despite these advances, more targets are needed to broaden the options available to patients. We tested a potent and specific protein arginine methyltransferase 5 (PRMT5) inhibitor in combination with BH3 mimetics for synergistic cell death in in vitro and in vivo. PRMT5 inhibition causes the upregulation of many pro-apoptotic members of the BCL-2 family of proteins, which are responsible for triggering intrinsic apoptosis. Mechanistic studies showed that response to PRMT5 inhibition and a combination treatment varied by cell line, but we saw significant survival advantages in our PDX models of MCL. PRMT5 inhibition with venetoclax provided the greatest advantage and disease was rendered undetectable via flow cytometry in two PDX models. Beyo (open full item for complete abstract)

Committee: Robert Baiocchi (Advisor); Sameek Roychowdhury (Committee Member); Monica Venere (Committee Member); Rosa Lapalombella (Advisor) Subjects: Biomedical Research

Doctor of Philosophy, The Ohio State University, 2016, Biomedical Sciences

Over the past decade, it has become clear that oncogenesis is a process driven by a wide variety of triggers including gene mutations, gene amplifications, inflammation, and immune deficiency. The growing pool of data collected from whole genome and epigenome studies of both solid and blood cancers has pointed toward dysregulation of chromatin remodelers as a unique class of cancer drivers. Next generation sequencing studies of lymphomas have identified a wide array of somatic mutations affecting enzymes that regulate epigenetic control of gene expression. Lymphoma is a type of cancer that originates in secondary lymphoid organs and manifests as an outgrowth of transformed lymphocytes, or white blood cells (WBCs) in the blood. The majority of lymphoma cases can be grouped into the Non-Hodgkin's lymphoma (NHL) subset and mainly occurs in B-cells. B-cell NHL is a heterogeneous set of cancers that would benefit from new therapies to improve patient progression-free survival. Cancers such as NHL typically present with a combination of genetic and epigenetic aberrations that contribute to the malignancy program. The epigenetic modifier protein arginine methyltransferase 5 (PRMT5) is required for B-cell transformation following Epstein-Barr virus (EBV) infection, and is overexpressed in various subsets of B-cell NHL. Based on these data we hypothesized that PRMT5 is a major driver of B-cell lymphomagenesis. To explore the role of PRMT5 in the development and progression of B-cell NHL we created a small molecule inhibitors targeted to PRMT5. Using the NHL subset mantle cell lymphoma (MCL) as a model we tested the efficacy of the drug. We discovered that PRMT5 was overexpressed in MCL primary samples and cell lines as compared to normal resting B cells. Furthermore, use of the small molecule inhibitor decreased the proliferation and viability in these cells without affecting the normal B-cells. Additionally, use of inhibitors caused G2/M cell cycle and decreased the express (open full item for complete abstract)

Committee: Robert Baiocchi (Advisor); David Lucas (Committee Member); Gregory Lesinski (Committee Member); Deepa Sampath (Committee Member) Subjects: Biology; Biomedical Research; Cellular Biology; Experiments; Health Sciences; Molecular Biology

Master of Sciences (Engineering), Case Western Reserve University, 2016, Biomedical Engineering

Non-Hodgkin's lymphoma, the most common type of blood cancer, has traditionally been treated with chemotherapeutics. The side effects of this treatment regime significantly affect quality of life for patients. As a result, researchers are developing targeted therapies that deliver the drug to the site of disease more efficiently; the mainstream of developments has focused on antibody-drug conjugates. As an alternative, nanoparticle drug therapies have shown great promise enabling drug delivery to sites of disease based on their size, shape and surface engineerability. Viral nanoparticles are biology-derived carriers that are growing in popularity due to their simple genetic and chemical modification, size tunability from spherical to high-aspect ratio, and biocompatibility. These attributes give a wide range of tools for engineers to design therapeutics with specific toxic loads and surface chemistries to efficiently navigate the body. In this thesis, I developed a viral nanoparticle using the nucleoprotein component from the tobacco mosaic virus (TMV) as the carrier for delivery of the anti-mitotic drug valine-citrulline monomethyl auristatin E (vcMMAE). I demonstrate successful synthesis of the formulation, and effective cell killing of Non-Hodgkin's lymphoma in vitro.

Committee: Nicole Steinmetz Dr. (Committee Chair); Giuseppe Strangi Dr. (Committee Member); Nicole Seiberlich Dr. (Committee Member) Subjects: Biomedical Engineering; Biomedical Research

Doctor of Philosophy, The Ohio State University, 2023, Veterinary Biosciences

Though cancer treatments have advanced significantly over the last few decades, drug resistance is responsible for 90% of cancer related deaths. Next generation sequencing technologies can characterize the molecular phenotypes of these patient tumors that evolve with resistance. In addition, these findings can help to inform the development of novel therapeutic strategies and combination therapies. While many cancers are susceptible to resistance, those with innately high degrees of genomic instability such as Mantle Cell Lymphoma (MCL) are most at risk. MCL is an incurable B-cell non-Hodgkin's lymphoma (NHL) and patients who relapse on targeted therapies have a poor prognosis. Protein arginine methyltransferase 5 (PRMT5), an enzyme essential for B-cell transformation, drives multiple oncogenic pathways and is overexpressed in MCL. Despite the anti-tumor activity of PRMT5 inhibition (PRT-382/PRT-808), drug resistance has been observed in several preclinical MCL models. Specifically, in the patient derived xenograft (PDX) MCL and double transgenic SOX11/CCND1 models, decreased survival of mice engrafted with these PRMT5 inhibitor resistant cells versus treatment-naive cells is observed (p-value: 0.005 and <0.0001, respectively). MCL cell lines also showed variable sensitivity to PRMT5 inhibition based on their half-maximal inhibitory concentrations (IC50s). Using PRT-382, cell lines were classified as sensitive (n=4; IC50 20-140 nM) or primary resistant (n=4; 340-1650 nM). Prolonged culture of sensitive MCL lines with drug escalation produced PRMT5 inhibitor resistant cell lines (n=4; 200-500 nM). This resistant phenotype persists after prolonged culture in the absence of drug and is observed with PRT-808. In the resistant PDX and cell line models, symmetric dimethylarginine (SDMA) reduction is achieved at the original PRMT5 inhibitor IC50 suggesting activation of alternative resistance pathways. Bulk RNA sequencing of resistant cell lines and PDX relative to se (open full item for complete abstract)

Committee: Robert Baiocchi Dr. (Advisor); Lapo Alinari Dr. (Advisor); Bradley Blaser Dr. (Committee Member); Samantha Evans Dr. (Committee Member); Christopher Oakes Dr. (Committee Member) Subjects: Biomedical Research

Doctor of Philosophy, The Ohio State University, 2023, Veterinary Biosciences

Antibody-based T-cell targeting immunotherapies, such as Immunotoxins (ITs) and monoclonal antibodies (mAbs), have demonstrated effectiveness in specifically depleting targeted T cells. These reagents have substantial benefits as precision medicines for T-cell lymphoma, organ transplant, autoimmune disease, and graft-versus-host disease (GVHD) and are currently being evaluated as a preconditioning reagent for cell therapy. CD3e-ITs, composed of a T-cell-binding peptide (e.g., CD3e antibodies) conjugated with toxins, have been developed to ablate a broad T-cell population. Recombinant human CD3e-IT, Resimmune, has demonstrated significant therapeutic benefit in cutaneous T-cell lymphoma (CTCL) with reduced toxicity. However, despite these promising results, there are still significant hurdles that need to be addressed for the successful application of CD3e-IT and T-cell targeting reagents in clinics. These hurdles include vascular leakage syndrome (VLS) and treatment resistance, as well as a poor understanding of the pharmacodynamics of CD3e-IT. To address the mechanisms underlying VLS development, we dissected and compared different portions of immunotoxins for VLS development and demonstrated that toxin molecules, not the T-cell binding portion, are the main driver of vascular leakage. Next, to understand the T-cell depletion efficacy of immunotoxins, we analyzed T-cell depletion in eleven different organs, including peripheral blood, spleen, five different locations of LNs (mesenteric, inguinal, mandibular, mediastinal, and lumbar LNs), Peyer's patches, bone marrow, thymus, and lung. We found that the treatment efficacy of CD3e-IT largely depends on the amount and quality of CD3e on the cell surface. CD3e surface expression is variable by different T subpopulations, and even the same subpopulations showed variable expression levels of CD3e depending on the organs in which the cells reside. In particular, the depletion rates of CD4+ FoxP3+ regulatory T cells (Treg) (open full item for complete abstract)

Committee: Sanggu Kim (Advisor); Prosper Boyaka (Advisor); Gregory Behbehani (Committee Member); Hazem Ghoneim (Committee Member) Subjects: Immunology

Master of Science, The Ohio State University, 2022, Comparative Biomedical Sciences

Inflammatory bowel disease (IBD), protein-losing enteropathy (PLE), and intestinal lymphoma (iLSA), encompass a large portion of gastrointestinal conditions in dogs presenting with chronic enteropathy (CE). The goal of this study was to determine whether gastrointestinal endoscopic evaluation or histopathologic diagnosis influenced clinician management of canine CE, particularly IBD, PLE, and iLSA. We hypothesized that endoscopic and histopathologic clinical data would affect clinician treatment decisions. Ten cases of IBD, 10 cases of PLE, and 9 cases of iLSA were retrospectively identified by histopathology reports. from the Ohio State University Veterinary Medical Teaching Hospital and Texas A&M GI Lab databases. IBD cases included lymphoplasmacytic inflammation in the duodenum, ileum, and/or colon. Exclusion criteria included other types of inflammation were present, presence of hypoalbuminemia, clinical suspicion for a differential diagnosis causing clinical signs, reasonable suspicion for lymphoma, or additional recommended testing. PLE cases included gastrointestinal inflammation with hypoalbuminemia. Exclusion criteria included a diagnosis of or reasonable suspicion for lymphoma or clinical suspicion for a differential diagnosis causing clinical signs or hypoalbuminemia. iLSA cases included a definitive diagnosis of lymphoma on histopathology alone or in combination with additional testing. Cases were excluded if additional testing was recommended but not performed. Each case was curated into 3 separate case presentations – case presentation 1 (CP1) with all case information excluding endoscopy and histopathology, case presentation 2 (CP2) with all case information excluding histopathology, and case presentation 3 (CP3) with all case information including both endoscopy and histopathology. These case presentations were randomized into 87 surveys and sent to 15 clinician evaluators blinded to the fact that these cases were curated from the original 29 (open full item for complete abstract)

Committee: Jenessa Winston (Advisor); James Howard (Committee Member); Valerie Parker (Committee Member); Adam Rudinsky (Committee Member) Subjects: Veterinary Services

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

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

Molecules that inhibit discrete protein-protein (PPIs) interactions hold immense promise to be developed as therapeutics or chemical genetics agents. The B cell lymphoma 2 (BCL-2) proteins are a family of evolutionarily-related proteins that act as positive or negative regulators of the intrinsic apoptosis pathway. Proteins in the BCL-2 family are primarily categorized into three sub types: anti-apoptotic (pro-survival) proteins, pro-apoptotic (pro-death) proteins and BH3-only proteins. Overexpression of anti-apoptotic BCL-2 proteins in cells is associated with apoptotic resistance, which can result in cancerous phenotypes or pathogenic cell survival. As a consequence, antiapoptotic BCL-2 proteins have attracted considerable interest as therapeutic targets. Antiapoptotic BCL-2 proteins bind to helical BH3 domains of pro-apoptotic proteins through a shallow, hydrophobic cleft. These pro-apoptotic proteins remain inactive when bound to anti-apoptotic members. BH3-only proteins are expressed under cell stress conditions to prevent interaction between ant-apoptotic BCL-2 and pro apoptotic proteins, which eventually leads to apoptosis initiation. Unfortunately, non-specific interactions between BCL-2 family members has made it difficult to elucidate specific mechanisms of BCL-2 signaling. Molecules that clearly define such interactions would be beneficial in the development of therapeutics that target specific proteins in the BCL-2 family. Scyllatoxin (ScTx) is a 31-amino acid protein isolated from scorpion venom that folds into α/β4 structural motif stabilized by three disulfide linkages, Cys3-Cys21, Cys8-Cys26 and Cys12-Cys28. Previous work by our lab has shown that ScTx can be engineered to target anti-apoptotic BCL-2 proteins by grafting residues required for Bcl-2 recognition to the α helix of ScTx. These constructs also showed that the number of disulfides within the ScTx BH3 mimetics influence favorable Bcl-2 recognition. Notably, ScTx-BH3 domain mimetics containi (open full item for complete abstract)

Committee: Justin Holub Prof (Advisor); Held Michael Prof (Committee Member) Subjects: Biochemistry; Biology; Biomedical Engineering; Biomedical Research; Biophysics; Chemistry

Doctor of Philosophy, Case Western Reserve University, 2019, Epidemiology and Biostatistics

Oncology clinical trials generate the evidence required to obtain regulatory approval for new interventions; the life-saving treatments cancer patients receive today, and the novel therapies that will transform future care paths, rely on data from clinical trials. Unfortunately, the therapeutic advances driven by clinical research are limited to the patient populations that best represent those enrolled in clinical trials. The explicit driver of this limited generalizability is the design of clinical trial eligibility criteria. Although overly restrictive eligibility criteria have been critiqued in the literature, quantitative studies evaluating the appropriateness of these criteria have not been performed. Therefore, we analyzed the eligibility criteria of a particular oncology disease group (hematologic malignancies), specifically exploring: 1) the relationship between commonly used organ function eligibility criteria and the expected toxicities of the trials' interventions, 2) reasons for ineligibility and the outcomes of leukemia patients eligible vs. ineligible for South Western Oncology Group (SWOG) trials, and 3) the health policy implications of overly restrictive eligibility criteria. Collectively, the findings of these studies suggest that the eligibility criteria for hematologic malignancy clinical trials are overly restrictive; the organ function criteria fail to reflect the expected toxicities of the trials' interventions / observed adverse events and the administrative criteria associated with the timing of screening tests / sample submissions included in SWOG leukemia protocols are too conservative. Furthermore, our results demonstrated the safety and efficacy outcomes were comparable between the leukemia/myelodysplastic syndrome patients ineligible for administrative or non-clinically significant reasons and the patients fully meeting the eligibility criteria. These findings suggest, patients who may benefit from potentially life-saving treatmen (open full item for complete abstract)

Committee: Siran Koroukian PhD (Advisor); Dana Crawford PhD (Committee Chair); J.B. Silvers PhD (Committee Member); Mikkael Sekeres MD, MS (Committee Member) Subjects: Design; Health Care; Health Sciences; Medicine; Oncology; Public Policy

Doctor of Philosophy, The Ohio State University, 2017, Comparative and Veterinary Medicine

Cutaneous T-cell lymphoma (CTCL) is a rare form of non-Hodgkin's lymphoma that is characterized by mature CD4+ T-cells that home to and proliferate within the skin. In the majority of patients, the disease eventually progresses to involve the blood and viscera. This is a heterogenous and complex cancer, and there are few effective therapies available for patients with advanced-stage disease. A hallmark of CTCL progression is epigenetic dysregulation, particularly involving abnormal expression of microRNAs (miRs). Novel therapeutic compounds with some promise in CTCL are epigenetic modifying drugs, such as histone deacetylase inhibitors (HDACi). However, despite initial responses in some patients, many will eventually relapse or develop clinical resistance. This failure highlights the urgent need for better understanding of the underlying pathogenesis of CTCL progression as well as identification of novel therapeutic targets. Therefore, we hypothesized that miRs play a central role in the pathogenesis of CTCL, by acting as both oncogenic and tumor suppressive factors, and that therapeutic targeting of miRs may represent an effective treatment strategy for CTCL patients. The first part of this thesis investigates an oncogenic miR, miR-181, in CTCL patient-derived cell lines as well as in primary CTCL patient cells. Previous work from our lab had shown decreased expression of sterile alpha motif (SAM) and histidine/aspartic acid (HD) domain-containing protein 1 (SAMHD1) in CTCL patients. SAMHD1 is a deoxynucleoside triphosphate hydrolase with established roles in nucleic acid metabolism, restriction of human immunodeficiency virus -1 replication, regulation of innate immune signaling, and modulation of DNA damage signaling. Recently, this gene has been implicated as a tumor suppressor gene in several types of cancer. The regulation of SAMHD1 and its potential role in the pathogenesis of CTCL was not previously known. We discovered that SAMHD1 protein levels were c (open full item for complete abstract)

Committee: Anjali Mishra (Advisor); Michael Caligiuri (Committee Member); Amy Johnson (Committee Member); Robert Baiocchi (Committee Member); Krista La Perle (Committee Member) Subjects: Biomedical Research

Doctor of Philosophy, The Ohio State University, 2017, Molecular, Cellular and Developmental Biology

MicroRNA-122 (miR-122) is a liver-specific microRNA that maintains liver homeostasis by regulating lipid metabolism, cell differentiation and viral infections in vertebrates. In clinics, hepatocellular carcinoma (HCC) patients with low miR-122 levels are usually associated with poor prognosis, implying miR-122 functions as a tumor suppressor. This notion is supported by the phenotypes of miR-122 genetic knockout (KO) mouse that develops spontaneous hepatitis, steatosis, fibrosis and HCC with age. Although several studies have shown the importance of miR-122 in maintaining liver homeostasis, the mechanisms by which loss of miR-122 contributes to these liver pathological processes remains largely unknown. In the current study, we explored the role of miR-122 in regulating liver inflammation and fibrosis by combining molecular, biochemical, and bioinformatic analysis. We demonstrated that the increased expression of the chemokine CCL2 in the liver is one of the causes of liver inflammation upon deprivation of miR-122. Blocking CCL2 using specific neutralizing antibody (CCL2 nab) ameliorates liver inflammation and tumorigenesis through decreasing the population of CD11b+/Gr1+ cells and their corresponding downstream pathways such as the IL-6-Stat3-cMYC axis and TNF-alpha-NF-kappaB axis. Along with the shrinking tumors in the CCL2 nab treated liver, CCL2 nab also activated natural killer (NK) cells and increased their cytotoxicity toward tumor cells. Besides its robust role in inhibiting liver inflammation and HCC tumors, miR-122 also has a strong anti-fibrosis function. Analyzing database that contained both clinical and expression profiles of liver cirrhosis patients revealed downregulation of miR-122 in the cirrhotic liver tissues compared to normal livers. Ectopic expression of miR-122 in LX-2, an immortalized human hepatic stellate cell (HSC) cell line, reduced cell proliferation and fibrotic gene expressions. Moreover, co-culture of miR-122 expressed HCC cells with (open full item for complete abstract)

Committee: Kalpana Ghoshal Ph.D (Advisor); samson Jacob Ph.D (Committee Member); Jianhua Yu Ph.D (Committee Member); David Brigstock Ph.D (Committee Member) Subjects: Biology; Molecular Biology; Oncology

Doctor of Philosophy, The Ohio State University, 2017, Comparative and Veterinary Medicine

Adult T-cell Leukemia/Lymphoma (ATL) is a rapidly fatal hematological malignancy that develops in a subset of Human T-cell Leukemia Virus Type 1 (HTLV-1) carriers after a 20-40 year latency. ATL has a unique relationship to bone characterized by aggressive bone invasion, long latency in the marrow, osteolytic bone metastasis, and hypercalcemia of malignancy. ATL bone metastasis and paraneoplastic syndromes cause substantial patient morbidity and a poor prognosis of less than a year. Novel therapeutic approaches for ATL are imperative to improve patient quality of life and prognosis. Therapies targeting both tumor and the bone microenvironment have been investigated in other bone tropic tumors but largely remain unstudied in ATL bone metastasis. This is due to the lack of understanding the mechanisms of the ATL tumor-bone communication and the paucity of appropriate animal models to study this focus of the disease. Several animal models exist to study the pathogenesis of HTLV-1 and ATL; however, models to recapitulate and describe the intimate tumor-bone interaction have not been well described. In this study, we developed and characterized novel xenograft mouse models of ATL bone metastasis. We introduced primary and in vitro HTLV-1 transformed cell lines into the tibias of immunosuppressed mice. Mice developed aggressive bone tumors and distant metastasis within 4 weeks. Tumor-bone phenotypes of xenograft mice ranged from minimal bone changes, mixed osteolytic/osteoblastic lesions, to marked osteolytic phenotypes similar to what is seen in clinical patients. The RVATL xenograft model with the greatest degree of tumor-associated osteolysis had significantly higher mRNA expression of parathyroid hormone-related protein (PTHrP), suggesting a significant paracrine role for PTHrP in ATL bone metastasis. Knockdown of PTHrP in an ATL cell line significantly decreased tumor growth and associated osteolysis in vivo. These models mimic key aspects of ATL in bon (open full item for complete abstract)

Committee: Thomas Rosol DVM, PhD (Committee Member); Maxey Wellman DVM, PhD (Committee Member); Beth Lee PhD (Committee Member); Toribio Ramiro DVM, PhD (Committee Member) Subjects: Comparative; Pathology

Doctor of Philosophy, The Ohio State University, 2016, Comparative and Veterinary Medicine

Around 12% of human cancers are caused by viral infections. Cancer-inducing viruses, called “oncoviruses”, hijack cellular machinery to avoid normal cell growth controls and detection of the infected cell by the immune system. Epstein-Barr virus (EBV), which infects over 90% of the world's population, is the most potent of these oncoviruses and is responsible for a diverse spectrum of malignant diseases. If the immune system becomes compromised, as in patients with AIDS or undergoing organ transplant treatment, the virus can initiate its growth program and transform host B cells into malignant lymphoma. These lymphomas are typically aggressive and exhibit poor clinical outcomes in an already-vulnerable patient population. Currently there is no standard treatment for EBV-associated lymphomas, demonstrating a significant unmet need for novel preventative and therapeutic approaches. Since nearly all individuals have been infected with EBV early in life, the immune system is already able to detect these EBV-infected cells. If properly activated and released from the suppressive effects of cancer, the immune system can use this previous detection, “immune memory”, to recognize and eliminate EBV-infected tumor cells in the setting of lymphoma. When EBV infects a cell, it uses the normal cell machinery to produce viral proteins that drive cell growth and survival signals. Dysregulation of these signals contributes to the emergence of lymphoma. In individuals with a normal immune system, these viral proteins are recognized by a type of white blood cell called cytotoxic T lymphocytes (CTLs) that kill infected cells to prevent viral spread. Here we show that a compound called silvestrol selectively targets B cell cancers while sparing normal immune cells by blocking the production of viral cancer-promoting proteins, mainly latent membrane protein 1 (LMP1), increasing immune cell-mediated death of cancer cells. Animal models of EBV-driven lymphoma treated with silvestrol s (open full item for complete abstract)

Committee: Robert Baiocchi MD, PhD (Advisor); David Lucas PhD (Committee Member); Aharon Freud MD, PhD (Committee Member); Michael Caligiuri MD (Committee Member) Subjects: Cellular Biology; Immunology; Virology

Doctor of Philosophy, The Ohio State University, 2016, Public Health

Various exposures have been investigated by epidemiologic studies as risk factors for leukemia incidence, however studies focusing on and therefore findings particular to Small Lymphocytic Lymphoma (SLL) and Chronic Lymphocytic Leukemia (CLL) have been sparse. In fact, according to the NCI, there are only a few established risk factors for CLL/SLL: 1) being middle-aged or older, male, or white; 2) a family history of CLL or cancer of the lymph system; and 3) having relatives who are Russian Jews or Eastern European Jews. As none of these are risk factors that can be altered with lifestyle changes, we sought to explore potential and likely risk factors that can be modified with behavior. Using the Women's Health Initiative (WHI) and applying an age and race matched, nested, 1:4 case-control design, we investigated CLL/SLL risk in postmenopausal women with three specific aims of interest: personal habits, comprising diet, drinking habits (including alcohol and coffee, both which have potential biological activity in leukemogenesis), and exercise; hormonal exposures, such as oral contraceptives (OC) and hormone therapies (HT); and pesticide exposures. These areas were chosen because there is a great need to understand 1) why CLL/SLL is significantly more prevalent in industrial countries compared to developing countries, and 2) why men have a two-fold increase in their risk of developing CLL/SLL. Women enrolled on WHI clinical trials who consumed coffee on a regular basis had lower risk of CLL/SLL (odds ratio (OR) = 0.73, 95% confidence interval (CI): 0.51, 1.05; P=.09), compared to non-coffee drinkers. Past oral contraceptive use (OR=0.74, 95% CI: 0.56, 0.96; P=.03) and obesity (OR=0.71, 95% CI: 0.53, 0.94; P=.20) both showed to be protective against CLL/SLL, whereas past estrogen use (OR=1.32, 95% CI:1.02, 1.71; P=.04) increased the risk. We did not find any significant associations with other personal habits and CLL/SLL risk, such as alcohol use, dietary factors (open full item for complete abstract)

Committee: Electra Paskett PhD (Advisor); Stanley Lemeshow PhD (Committee Member) Subjects: Epidemiology

MS, University of Cincinnati, 2012, Medicine: Clinical and Translational Research

Purpose: To identify associations between specific therapy-related events such as length of hospitalization and neuropathic pain with health-related quality of life (HRQOL) outcomes for children with acute lymphoblastic leukemia and lymphoblastic lymphoma. Hypothesis: In children (ages 2 – 30 years) with acute lymphoblastic leukemia (ALL) or lymphoblastic lymphoma, poor health-related quality of life (HRQOL) scores as measured by the PedsQL modules during low-intensity “maintenance” therapy are significantly associated with increased care measures during the initial phases of therapy, including prolonged hospitalization and use of narcotics or neuropathic pain medications. Specific Aims: 1.To evaluate the relationship between extended hospitalization time (as defined by total days or weeks admitted, or by number of unanticipated admissions) during intensive phases of therapy, and HRQOL scores during maintenance therapy in children ages 2 years and older with ALL or lymphoblastic lymphoma. 2. To evaluate the relationship between increased supportive care for pain by use of patient-controlled analgesia (PCA), long acting oral narcotics, or the use of neuropathic pain medications during therapy, and HRQOL scores during maintenance therapy in children ages 2 years and older with ALL or lymphoblastic lymphoma. Methods: A cross-sectional single institution pilot study was performed for children with ALL or lymphoblastic lymphoma who had reached maintenance therapy or had recently completed treatment. HRQOL scores were obtained using the PedsQL 4.0 Generic Core Scales and PedsQL 3.0 Cancer Module, and retrospective chart review was performed for each patient. Results: 57 subjects were available for analysis. Multiple regression analysis found that longer total time of hospitalization correlated with poorer overall PedsQL 4.0 summary scores (p = 0.002). Age, time since the start of maintenance therapy, and the number of unscheduled hospitalizations were independently progno (open full item for complete abstract)

Committee: Erin Nicole Haynes DrPH (Committee Chair); Jareen Meinzen-Derr PhD MPH (Committee Member); Karen Burns MD MS (Committee Member) Subjects: Oncology

MS, University of Cincinnati, 2010, Engineering and Applied Science: Mechanical Engineering

Primary intraocular lymphoma (PIOL) is a term that refers to non-metastatic malignant lymphoid neoplasia that arises primarily within the eye. One of the currently available treatment options for PIOL is intravitreal injection of methotrexate. The main objective of this study is to numerically investigate the kinetics of methotrexate in a human eye following the standard induction–consolidation–maintenance (I-C-M) injection protocols for the treatment of PIOL (numerical study). In addition, we investigated the intravitreal concentration of methotrexate over time, and toxicity associated with slow sustained release of the drug from a biodegradable device containing methotrexate implanted in a deep scleral pocket of the eyes of normal rabbits (in vivo study). The 3-dimensional model of a rabbit eye was adapted from our previous studies and a human eye model was constructed based on its physiological dimensions. The retinal permeability values of methotrexate calculated for an albino rabbit and human eye were 1.1×10-5 cm/s and 9.25×10-6 cm/s, respectively. Given that the dosage above 0.1µM is considered tumoricidal, the cytotoxic levels of methotrexate were consistently achieved only in the induction phase and the concentration levels dropped below the cytotoxic levels for part of the consolidation and maintenance phases of the treatment following the I-C-M protocols. By making use of the results of this study, one could select a dosing interval for serial injections of methotrexate or establish a treatment schedule using controlled release methotrexate implant for the treatment of PIOL. For the in vivo study, biodegradable micro-needle implants (approximately 8 mg) loaded with 10% wt of methotrexate were fabricated using solvent cast method. The implants were all appropriately rigid at room temperature or when cooled in a refrigerator following fabrication. The micro-needles were expected to penetrate the inner scleral lamellae in the scleral pocket and the un (open full item for complete abstract)

Committee: Rupak Banerjee PhD, PE (Committee Chair); Frank Gerner PhD (Committee Member); James Augsburger MD (Committee Member); Zelia Correa MD, PhD (Committee Member) Subjects: Mechanical Engineering

PhD, University of Cincinnati, 2007, Engineering : Mechanical Engineering

Intraocular lymphoma (IOL) needs a long term therapy and its treatment protocol has not been well established. A controlled drug release micro-implant, implantable in scleral pocket in the pars plana region of the eye, is considered to treat IOL as it allows sustained release of therapeutic levels of drug for a desired period of time; thus, overcoming the blood-retinal barrier associated with systemic drug delivery. Methotrexate (MTX) is the most commonly used chemotherapeutic agent to treat IOL. 2-Methoxyestradiol (2ME2) is a potent antitumor and antiangiogenesis agent which, unlike other cytotoxic drugs, has minimal toxicity. To establish a treatment method using controlled release implant, 1) therapeutic range of 2ME2 is evaluated by comparing with MTX in proliferation study with human lymphoma cell line; 2) biodegradable microneedle implant is fabricated to obtain sustained release of 2ME2 within the therapeutic range; 3) computational eye model is created to calculate drug distribution and pharmacokinetics. Results show that the drug release rate of the 25% w.t. 2ME2 microneedle implant follows zero order kinetics without significant burst having mean release rate of 0.36 microg/hr over a 90 days period. The efficacy studies of 2ME2 show dose-time dependent growth inhibition of lymphoma cell lines within the range of 0.1 ~ 1 microM. It is expected that the 25% w.t. 2ME2 microneedle implant has 0.36 microM mean vitreous concentration in the human eye. The implant can sustain within the therapeutic range of 2ME2 on lymphoma cell line (0.1 – 1 microM) for around 90 days. Pharmacokinetic evaluation using computational eye model show that the half-life of the MTX in the vitreous is 7.1 hr for the rabbit eye and 13.9 hr for the human eye. In conclusion, sustained-release of 2ME2 using intravitreal biodegradable micro-needle implants, designed to deliver therapeutic levels of 2ME2 for an extended period of time, shows promise for the treatment of IOL.

Committee: Dr. Rupak Banerjee (Advisor) Subjects:

MS, University of Cincinnati, 2006, Medicine : Epidemiology (Environmental Health)

This study examined the association between chronic residential exposure to low-level ionizing radiation and risk of leukemia and non-Hodgkin's lymphoma (NHL). Subjects included 8,220 people living within a five-mile radius of a uranium processing plant in Fernald, Ohio. A retrospective cohort design was used, including incident cases of leukemia and NHL that developed after 1990. Estimates of radiation exposure were based on subjects' location and duration of residence. Logistic regression and Cox proportional hazards regression were used to determine if there was an association between level of radiation exposure and risk of leukemia and NHL, controlling for age. Other covariates included gender, SES, smoking status, X-ray history, family history of cancer, and exposure to benzene, asbestos, or farming. Results indicated a non-significantly elevated OR for radiation exposure among the leukemia cohort. For the NHL cohort, there was no clear association between radiation exposure and risk of disease.

Committee: Dr. Susan Pinney (Advisor) Subjects: Health Sciences, Public Health