Biomedical Science Undergraduate Research Theses and Honors Research Theses

Permanent URI for this collection

Undergraduate Research Theses and Honors Research Theses from the School of Biomedical Science

Instructions for students

Browse

Recent Submissions

Now showing 1 - 20 of 74
  • Item
    Interactions Between PIK3CA and MAP3K1 Pathways in Breast Cancer
    (The Ohio State University, 2024-05) Meadows, Owen; Toland, Amanda
    Introduction/Background: Breast cancer is one of the leading causes of death in the United States, with one in eight women being diagnosed in their lifetimes. Many tumors are susceptible to treatment targeted towards their specific molecular subtypes. One such biomarker based on molecular subtype is the human epidermal growth factor receptor 2 (HER2). HER2-positive tumors historically have been associated with poorer outcomes, but in recent years targeted therapies have significantly improved prognoses. However, many tumors develop resistance to drugs targeting those molecules. One study of interest found that in patients treated with anti- HER2 agent trastuzumab, individuals harboring PIK3CA mutations experienced markedly reduced progression free survival following treatment compared to wild type patients. PIK3CA also displays interactions with the MAPK signaling pathway. Our laboratory found that nucleotide variants (SNVs) at the MAP3K1 locus associated with PIK3CA mutations in breast tumors. Understanding the link between PIK3CA and MAP3K1 may help to develop more individualized treatments for patients with HER2+ tumors or other PIK3CA mutated tumors. Methods: Isogenic MCF10A and 12A cell lines with wild-type (WT) and PIK3CA H1047R knock-in (KI) variants were used as in vitro models of breast cancer. MAP3K1 expression was knocked down through transfection, utilizing siRNAs to create cells deficient in MAP3K1. MTT assays were performed to analyze differences in proliferation between cellular conditions. Protein was extracted for western blotting to confirm MAP3K1 knock-down, using GAPDH as a control. Results: Western blot analysis of transfected cell lines confirmed that siRNAs were successful in knocking down expression of the MAP3K1 protein. MTT analysis showed that knocking down MAP3K1 resulted in increased fold growth in PIK3CA KI cells compared to WT. Data collection and analysis in other cell lines for greater validity is currently still ongoing. Conclusions: Based on the results of the MTT assays, initial studies indicate MAP3K1 expression has a negative correlation vs PIK3CA mutation on tumor growth. Cells with lower levels of MAP3K1 protein and PIK3CA mutations showed increased proliferation. Further investigation of these two pathways could help to better understand how they influence each other.
  • Item
    Inhibition of TGF-β/Smad signaling attenuates extracellular matrix accumulation in a human ex-vivo model of pulmonary fibrosis
    (The Ohio State University, 2024-05) Riley, Madeline; Rojas, Mauricio; Rosas, Lorena
    Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive lung disease characterized by accumulation of scar tissue and decreased lung function. Mechanisms of disease onset have yet to be fully understood. However, activation of transforming growth factor-beta (TGF-β)/Smad signaling, followed by repeated injury to the epithelium, results in transdifferentiation of fibroblasts into profibrotic fibroblasts, leading to an increase in collagen expression and accumulation in the extracellular matrix (ECM), resulting in deteriorating lung function. Therefore, we proposed a TGF-β/Smad inhibitor to prevent ECM accumulation. We used human precision cut-lung slices (hPCLS) treated with TGF-β as a human ex-vivo model to retain native lung tissue structure, cellular complexity, and ECM protein composition. hPCLS were stimulated for 48 h with active recombinant human TGF-β (10ng/ml). Then, a TGF-β/Smad inhibitor (SB431542, 10µM) was added to the culture for 72 h and hPCLS were later harvested and analyzed to evaluate the expression of ECM proteins by immunofluorescence and PCR. We found high expression of collagen 1a1, collagen 3a1, fibronectin, and α-SMA after TGF-β stimulation. Treatment with the TGF-β/Smad inhibitor attenuated their expression. Our data suggests that the ex vivo model of hPCLS can be used to evaluate anti-fibrotic therapies. We demonstrated that specific inhibition of TGF- β results in the attenuation of the expression and deposition of proteins of the ECM. These results corroborate that modulation of TGF-β activity using SB431542 inhibitor has a therapeutic potential for patients with pulmonary fibrosis.
  • Item
    Analysis of African American Patients Diagnosed with Acute Promyelocytic Leukemia Unveils Comparable Survival and Unique Genomic Characteristics
    (The Ohio State University, 2024-05) Boateng, Isaiah; Eisfeld, Ann Kathrin
    Among the various hematological malignancies Acute Promyelocytic Leukemia (APL) represents a distinct subtype of acute myeloid leukemia (AML), characterized by the PML/RARa fusion protein resulting from the translocation t(15;17).1 Despite advancements, there remains a gap in knowledge or in other words a comprehensive understanding of the molecular landscape of APL across diverse racial backgrounds. This study sought to investigate this gap by analyzing survival outcomes and genomic profiles of APL patients, particularly those of African descent. Using nationwide registries and clinical trial databases, survival analyses revealed no racial disparities in overall survival (OS) but highlighted age and social deprivation as significant prognosticators to OS. Molecular profiling of African American patients characterized the molecular landscape and uncovered both known and novel mutations including FLT3, CALR, and NLGN2, suggesting potential ancestry-related differences in driver mutations. Furthermore, analysis of gene fusions revealed variations in isoforms, indicating possible ancestry-associated differences in disease biology. These findings underscore the importance of inclusive genomic studies for a more complete understanding of molecular features and associated disease biology and highlight the value of comprehensively understanding the disease landscape, particularly in patient populations traditionally underrepresented. Moreover, it depicts, on a population-based level, just how impactful access to care is for patient survival and outcomes.
  • Item
    Characterizing Cancer Mutational Signatures using Computational Approaches
    (The Ohio State University, 2024-05) Nelakurti, Devi Dheekshita; Petreaca, Ruben C.; Mirzaei, Golrokh
    Cancers are characterized by accumulation of genetic mutations in key cell cycle regulators that alter or disable the function of these genes. Such mutations can be inherited or arise spontaneously during the life of the individual. The MEN1 gene prevents uncontrolled cell division and it is considered a tumor suppressor. Inherited MEN1 mutations are associated with certain parathyroid and pancreatic syndromes while spontaneous mutations have been detected in cancer cells. We investigated whether inherited mutations appear in cancer cells which would suggest that patients with parathyroid and pancreatic syndromes have a predisposition to develop cancer. Moving on, thousands of cancer genomes are now publicly available which has led to new insights into identification of mutational signatures at both nucleotide and amino acid levels. Here, we discuss C > T transitions as a key nucleotide-level mutational signature that leads to a dramatic overrepresentation of arginine substitutions in cancers. Beyond small-scale mutational signatures, we also explored the impact of chromosomal instability and large-scale mutations like copy number variations (CNVs) in DNA damage repair. We find a weak correlation between the spectrum of inherited mutations and those appearing spontaneously. Thus, inherited MEN1 mutations may not be a good predictor of tumorigenesis. We also propose that the underlying C > T mutational signature canalizes possible arginine substitution outcomes, favoring histidine, cysteine, glutamine, and tryptophan. This initial asymmetry is then acted on at the amino acid level by purifying selection. Thus, a model of “sequential selection” could explain the documented bias towards arginine substitutions in multiple cancers. Our investigation also reveals that mutations in RAD52, CTIP, and PRMT5 lead to intrachromosomal deletions affecting cell cycle enzymes, shows significant correlations among various gene interactions during DNA damage repair. By using computational methods, we are able to consolidate and find new associations and patterns in cancer signatures can be identified to obtain a more holistic map of cancer driver mechanisms.
  • Item
    Non-viral Direct Cell Reprogramming Methods Aiming to Rescue Glycemic Control in Type 1 Diabetes
    (The Ohio State University, 2024-05) Gotschall, Andrew; Gallego-Perez, Daniel
    Allogenic islet transplantation is the current gold standard cell therapy to treat patients with severe type I diabetes. However, its implementation is limited due to a lack of donor tissue, its invasive nature, and the requirement for patients to take immunosuppressants for the remainder of their lives following treatment. To address these limitations, we primarily investigated a novel method to directly reprogram fibroblasts into induced β cells. We delivered plasmid DNA encoding for a unique combination of three skin plasticity and seven β cell patterning transcription factors (3SP+7βC) to dermal fibroblasts and skin tissue using bulk electroporation (in vitro), nanoelectroporation (in vitro), or tissue nanotransfection (in vivo). Mouse embryonic fibroblasts transfected with the 3SP+7βC combination demonstrated increased insulin 1 and insulin 2 gene expression by day 14 compared to control. Moreover, these populations of mouse embryonic fibroblasts transfected with the 3SP+7βC combination stained positive for the insulin protein. In vivo studies suggest upregulated insulin gene expression, as well as acute glycemic control rescue, and are currently ongoing. Further experiments are still needed to explore the extent of induced β cell reprogramming and the glycemic control that can be driven by the delivery of unique transcription factors using electroporation-based methods. However, this study explores a step towards providing an alternative cell source to treat, and potentially cure, the lack of insulin causing homeostatic glucose imbalance in type I diabetes. Success here has potential to greatly improve diabetic patients' quality of life and relieve the financial burden faced by patients as they monitor type I diabetes throughout their lives. These improvements would also lower the financial burden faced by healthcare systems due to decreased demands for type 1 diabetes related micro-, and macrovascular disease treatments.
  • Item
    Inhibition of VWF Demonstrates Thrombolysis Following ex vivo Thrombus Formation in Patients with AIS LVO
    (The Ohio State University, 2024-05) Holthaus, Blake; Nimjee, Shahid
    Acute ischemic stroke (AIS) is the leading cause of combined death and disability across the world. While recombinant tissue plasminogen activator (rTPA) and its analogs are the sole approved thrombolytic for AIS, its application is constrained by its strict administration window due to lack of reversibility, and potential for hemorrhage. Von Willebrand factor (VWF) is a glycoprotein pivotal in the adhesion, aggregation, and propagation of platelets. Most notably, VWF is responsible for tethering a thrombus to the collagen within a vessel wall through interaction with the platelet glycoprotein receptor, GP-Ib. This interaction also further activates surrounding platelets and leads to deceleration in cerebral blood flow, exacerbating ischemic injury. Thus, VWF is a crucial mediator in AIS pathophysiology. BB-031 is an RNA aptamer shown to inhibit VWF-platelet interaction in in vivo and in vitro settings. The objective of this study was to determine if BB-031 will be an effective therapeutic in AIS large vessel occlusion (LVO) patients using a kinetic ex vivo microfluidic model. A microfluidic model of arterial thrombus was used to recapitulate the constant pressure and kinetic environment found with in vivo occlusion. After thrombus formation, BB-031 was introduced to the flow at a concentration of 1692 nM. A 2-hour perfusion period was observed following treatment administration of BB-031 or vehicle control (PBS). Patency and thrombus surface area were measured throughout the perfusion. In the kinetic model, patency was achieved in 85% (6/7) of samples in the BB-031 samples compared to 0% (0/8) of the samples in the vehicle control group. Analysis of the surface area of the thrombi showed a decrease in pre-occlusion platelets throughout the perfusion in the BB-031 treatment group compared to PBS control. Interestingly, post-occlusion platelets initially increased in surface area as new platelets were deposited and eventually decreased in the BB-031 group. The control group showed decreased post-occlusion platelet deposition. This study demonstrates the potential of BB-031 as a therapeutic intervention for large vessel occlusion acute ischemic stroke, exhibiting encouraging trends in enhancing patency and modulating thrombus formation. These results offer promising for those affected by this lethal disease and more work must be done to further strengthen the thrombolytic replacement of rTPA with BB-031 or at least, efficacy for those patients not eligible for therapeutic treatment.
  • Item
    NOD2-mediated monocyte/macrophage stimulation to enhance antibody therapies
    (The Ohio State University, 2024-05) Bull, Mikayla; Tridandapani, Susheela
    Chronic lymphocytic leukemia (CLL) is a malignant disease where mature B cells accumulate in the blood, as well as in lymphoid organs. Over 20,000 new cases and nearly 4,500 deaths are expected in the United States alone for 2024. Monoclonal antibody therapies are used to treat CLL, among other diseases. Antibodies bind monocyte/macrophage Fcγ receptors (FcγRs), inducing phagocytosis and inflammatory cytokine release to promote clearance of target cells. However, monocytes and macrophages are suppressed by CLL cells. Toll-like receptor activation has improved therapeutic efficacy in vivo, with some ligands even clinically approved. Here, we investigated the potential of nucleotide-binding oligomerization domain-containing protein 2 (NOD2) in this capacity, for a NOD2 agonist is approved by the European Medicines Agency for osteosarcoma treatment. NOD2 is an intracellular pattern recognition receptor, which, upon binding muramyl dipeptide (MDP), induces inflammatory cytokine production via the NF-κB and MAPK pathways. Using healthy-donor monocytes as a model, we first used ELISA to confirm activation after L18-MDP treatment (MDP derivative), as measured by increased TNF-α release. Using qPCR, we then detected upregulation of the activating FcγRIIa, IIIa, and associated γ-chain. We subsequently confirmed protein-level increase in γ-chain expression by western blot. Investigating FcγR-mediated functions, while L18-MDP did not significantly increase antibody-mediated cytokine release, we detected significantly greater phagocytic activity after NOD2 stimulation and FcγR engagement. Finally, by applying pharmacological inhibitors, we discovered varying levels of involvement of NF-κB, MEK, and p38 in NOD2-mediated modulation of FcγR expression. In conclusion, this study suggests that NOD2 agonists may be considered as immunostimulatory agents for use in combination with monoclonal antibody therapies for diseases such as CLL.
  • Item
    Characterizing Fibrotic Tissue in Duchenne Muscular Dystrophy Cardiomyopathy
    (The Ohio State University, 2022-05) Cumby, Laurel; Rafael-Fortney, Jill
    Duchenne Muscular Dystrophy (DMD) is an X-linked recessive disease caused by the lack of a functional dystrophin protein, which anchors the muscle cell cytoskeleton to the extracellular matrix. DMD is characterized by muscle degeneration leading to wheelchair use by the age of 12, use of ventilation assistance by the age of 20, and a progression into cardiac failure, leading to an approximate lifespan of 25 years. Prior DMD animal models were inadequate for studying cardiac failure, prompting the development and subsequent characterization of the Fiona/dko model, a mouse line that is dystrophin deficient, utrophin deficient, and possesses a human utrophin transgene expressed only in skeletal muscles, but not the heart. Investigation into the Fiona/dko model illustrated a gap in knowledge, as it was found that while cardiac function continues to decline through 12 months of age, quantity of fibrotic tissue plateaus after 6 months. Micro-dystrophin-5 gene therapy, currently being tested in clinical trials, was shown to prevent cardiac functional decline and pathology in Fiona/dko mice. To determine the cause of declining function with sustained fibrosis and subsequently locate therapeutic targets, it is necessary to further characterize fibrosis composition in the Fiona/dko model. Investigation of the extracellular matrix components comprising cardiac scars in Fiona mice was completed through immunofluorescence staining for Fibronectin, Elastin, Tenascin, and Collagen I, as well as staining of activated myofibroblasts using alpha-smooth muscle actin. A significant reduction of fibronectin and tenascin was present in treated Fiona/dko mice as compared to untreated Fiona/dko. Collagen I and Fibronectin localization was compared and shown to be localized to the same areas of fibrosis. Elastin was not found to be present in fibrotic scars of Fiona/dko hearts. Staining for alpha-smooth muscle actin was not prominent compared to other dystrophic tissues such as diaphragm, suggesting myofibroblast in dystrophic hearts may have a different phenotype. Immunostaining of extracellular matrix proteins suggest that their contribution to fibrotic scars does not change during heart failure progression and that micro-dystrophin gene therapies are able to prevent cardiac fibrosis in dystrophic hearts.
  • Item
    Stimulation of NOD2 in Acute Myeloid Leukemia activates Natural Killer Cells and improves survival
    (The Ohio State University, 2023-05) Murugesan, Rakesh; Tridandapani, Susheela
    Acute Myeloid Leukemia (AML) is a hematological malignancy of the myeloid lineage of immune cells in the blood and bone marrow. It is characterized by low five-year survival rates of 27.4%, and high incidence of relapse, indicating a need for novel therapeutic strategies. A signature of AML is the evasion of immune surveillance by Natural Killer (NK) cells, which are immune cells possessing anti-leukemic capabilities. NK cell activating receptors are downregulated in AML-NK cells, and these cells exhibit defective cytotoxic ability. Recently, our lab has found that treatment with MTP-PE (a synthetic ligand for NOD2 used for the treatment of osteosarcoma) along with IFNγ provided a survival benefit in a murine model of AML. Data from this study show that blood-derived NK cells isolated from mice treated with MTP-PE and IFNγ show greater levels of maturation, as indicated by markers CD27 and CD11b. Additionally, we saw increased expression of activation/degranulation markers, CD69 and CD107a, on healthy donor NK cells in in vitro cocultures with AML patient samples. This study is focused on elucidating the mechanism by which NK cell activation occurs when AML is treated with MTP-PE and IFNγ. Preliminary data suggests that ligand-receptor cell-to-cell contact between AML cells and NK cells is necessary to elicit the observed NK cell activation. In this study, we show that MTP/IFNγ treated AML cells strongly upregulate IL-15, an important cytokine involved in NK cell expansion and activation, as well as ligands of the receptor NKG2D. In AML patient samples, we have found the co-treatment to upregulate CD69 and CD107a on suppressed patient NK cells, both in peripheral blood and apheresis. We also observed expansion of NK cells in these patient samples. Further study is necessary to evaluate and understand the potential of MTP-PE and IFNγ to activate suppressed NK cells in Acute Myeloid Leukemia and improve anti-leukemic immunosurveillance.
  • Item
    Inhibition of Ras-like Protooncogenes A and B, RALA & RALB, Sensitize Soft Tissue Sarcomas to Radiation Therapy
    (The Ohio State University, 2023-05) Chakravarthy, Prathik; Sizemore, Steven
    Treatment options for patients with soft tissue sarcomas (STS) have changed very little over the past 50 years. While radiation treatment (RT) improves outcome for patients with many types of cancer, STS are notoriously RT resistant. Understanding the mechanism(s) of this resistance and how to overcome it will improve outcomes for STS patients. Recent approaches to understand radiation resistance in cancers have placed an emphasis on uncovering key genetic alterations that are helping to confer this radiation resistance. RALA and RALB are small GTPases that are differentially expressed in a variety of cancer and have been shown to play a role in conferring radiation resistance. In STS the RALs are also altered in expression making them promising targets to study the issue of radiation resistance in STS. The role of RALA and RALB in STS radiation resistance were investigated through in vitro assays studying cellular response to radiation with variable expression of RALs. Radiation responses analysis included cell survival and sustained DNA damage. RALA and RALB expression was experimentally decreased in STS cell lines to study the roles of the RALs during RT. Colony forming assays were performed to understand how the survival and growth of STS cells were altered with and without the expression of RALA and/or RALB. Additionally, DNA damage, one of the major responses to RT, was analyzed post-radiation in STS cells in which RAL expression was once again experimentally decreased. Assays involved immunofluorescence methods measuring the presence of yH2AX, a biomarker representative of DNA damage. Knockout or depletion of the RAL small GTPases inhibited the radiation resistance of STS cancer cells. Clonogenics assays showed that in vitro when the RALs were depleted there was a decrease in the surviving fraction of STS cells after radiation, particularly when RALB was depleted. This showed that increase in RAL could be responsible for radiation resistance in STS and inhibiting RAL expression could re-sensitize cells to RT. DNA damage post-radiation showed similar results with RAL depletion resulting in increased sustained DNA damage after radiation exposure. This once again shows that when RAL expression was experimentally decreased there was a more effective radiation response as the cells sustained a greater amount of DNA damage. Together these data show the significant role that RALs play in conferring radiation resistance to STS and perhaps a possibility in inhibiting RAL expression to sensitize STS cells to RT. Further investigation will be necessary to understand the mechanisms by which RALA and RALB are conferring this resistance for the eventual use of RALs as targets for more effective RT.
  • Item
    Cell-type specific transcriptomic signatures of neocortical circuit organization and their relevance to autism
    (The Ohio State University, 2023-05) Moussa, Anthony; Wester, Jason
    A prevailing challenge in neuroscience is understanding how diverse neuronal cell types select their synaptic partners to form circuits. In the neocortex, major classes of excitatory projection neurons and inhibitory interneurons are conserved across functionally distinct regions. There is evidence these classes form canonical circuit motifs that depend primarily on their identity; however, regional cues likely also influence their choice of synaptic partners. We mined the Allen Institute's single-cell RNA-sequencing database of mouse cortical neurons to study the expression of genes necessary for synaptic connectivity and physiology in two regions: the anterior lateral motor cortex (ALM) and the primary visual cortex (VISp). We used the Allen's metadata to parse cells by clusters representing major excitatory and inhibitory classes that are common to both ALM and VISp. We then performed two types of pairwise differential gene expression analysis: (1) between different neuronal classes within the same brain region (ALM or VISp), and (2) between the same neuronal class in ALM and VISp. We filtered our results for differentially expressed genes related to circuit connectivity and developed a novel bioinformatic approach to determine the sets uniquely enriched in each neuronal class in ALM, VISp, or both. This analysis provides an organized set of genes that may regulate synaptic connectivity and physiology in a cell-type-specific manner. Furthermore, it identifies candidate mechanisms for circuit organization that are conserved across functionally distinct cortical regions or that are region dependent. Finally, we used the SFARI Human Gene Module to identify genes from this analysis that are related to risk for autism spectrum disorder (ASD). Our analysis provides clear molecular targets for future studies to understand neocortical circuit organization and abnormalities that underlie autistic phenotypes.
  • Item
    Inhibition of BET proteins by novel inhibitor, PLX51107, regulates Fc-gamma receptor function and reduces inflammation in rheumatoid arthritis
    (The Ohio State University, 2021-05) Shankar, Divya; Tridandapani, Susheela
    Members of the Bromodomain Extra-Terminal Domain (BET) family — BRD2, BRD3, BRD4, and BRDT — are key regulators of transcription. BET inhibition is associated with reduced immune responses, making the BET family a potential therapeutic target for autoimmune diseases, including rheumatoid arthritis (RA). Autoimmune disease pathogenesis is dependent on the overactivation of immune responses. This includes the heightened production of inflammatory cytokines, like TNFα, and secretion of autoantibodies, such as isotypes of rheumatoid factor and anticitrullinated protein antibody (ACPA), which target healthy tissues. Fcγ receptors (FcγR) expressed on the surface of myeloid cells bind Immunoglobulin G (IgG) immune complexes; FcγRIa, FcγRIIa, FcγRIIIa are activating FcγR, whereas FcγRIIb is an inhibitory FcγR. Recognition of autoantibodies by FcγRs induces an inflammatory phenotype that results in tissue damage and further escalation of the inflammatory response. Here we examine a novel BET inhibitor, PLX51107, and its effect on regulating FcγR expression and function in RA. PLX51107 substantially upregulated FcγRIa and downregulated FcγRIIa, FcγRIIb, FcγRIIIa, and FcϵR1-γ (γ-chain) — FcγRIa and FcγRIIIa's associated signaling protein — mRNA and protein levels in healthy donor and RA monocytes. Consistent with this, PLX51107 treatment decreased phagocytic and rosetting indexes, FcγR-mediated NF-κB, AKT, and MAPK signaling, and TNFα production, indicating the attenuation of the immune response. Our findings suggest that BET inhibition may have clinical relevance in the treatment of RA. Further studies are critical to explore how BET inhibition mechanistically regulates FcγRs to alleviate their mediated inflammatory responses, and to determine if BET inhibition, via PLX51107, is therapeutic in a murine model of RA.
  • Item
    An Evaluation of Porosity and Bone Mineralization and Relationships with Fracture Risk in Human Tibiae
    (The Ohio State University, 2022-05) Goldsmith, Carla; Harden, Angela
    Bone quality assessment tools in clinical settings are used to evaluate fracture risk in living patients. Additionally, ex-vivo analyses of individual bones have assessed bone quality through examination of porosity and mineralization.1–3 However, a comprehensive understanding of relationships between demographics (e.g., age, sex, body size) and variables associated with bone quality is lacking. Understanding the factors contributing to bone quality will improve diagnosis of osteoporosis, a disease characterized by bone loss, and identify high fracture risk populations (e.g., older individuals, females). The objective of this study was to investigate the relationships between porosity and mineralization in human tibiae with sex, age, body size, and bone mineral density (BMD) and assess differences in bone quality assessment via methodology (clinical vs histological vs compositional). Sections from human tibiae were collected at the 66% site (n = 14) and the 15% site (n = 16) for porosity and mineralization analyses, respectively. Areal (aBMD) and volumetric (vBMD) BMD, collected via dual x-ray absorptiometry and quantitative computed tomography scans, values were compared to porosity, mineralization, and individual-level variables (e.g., sex, age, body size) to determine if these variables had relationships with these clinical bone quality assessment tools. Overall, few relationships were observed within this study; however, males (M=1.11 g/cm2) exhibited greater aBMD than females (M=0.93 g/cm2), mineralization had a negative correlation with age, and height and weight had positive correlations with vBMD. Future research should investigate porosity and mineralization utilizing a larger sample size, wider age range, and more varied body sizes per sex to better evaluate bone quality. Overall, a variety of bone- and individual-level variables should be used to make accurate assessments of bone quality to inform individuals' fracture risk.
  • Item
    Optimization of Novel Lipid Nanoparticle Formulations for mRNA Delivery
    (The Ohio State University, 2022-05) Bohmer, Margaret; Dong, Yizhou
    mRNA research has heightened in prevalence due to the emergence of mRNA-based vaccines for the SARS-CoV-2 virus, but there is still a need for carriers to deliver future mRNA-based vaccines and therapies. The aim of this study was to develop and optimize formulations for novel lipid nanoparticles (LNP) to deliver mRNA in different cell lines. One novel LNP, CW51, was selected for formulation optimization via an orthogonal array. These optimized formulations were able to transfect JAWS II dendritic and C2C12 skeletal muscle cells with luciferase mRNA. These findings indicate that the lead LNP warrants further investigation into its potential in mRNA-carrying nanoparticles.
  • Item
    Assessing the Impact of Adding Scenario-based Augmented Reality Training to Traditional Textbook Training on Accurately Identifying Airway Obstruction Symptoms
    (The Ohio State University, 2020-05) Mansour, Lauren; Patterson, Emily
    Recent developments in the portability, flexibility, and affordability of augmented reality (AR) technology allow it to address some limitations of current medical training. This pilot study aims to determine whether AR technology can enhance the detection and identification of subtle cues used to make accurate medical diagnoses. We randomly divided five participants into a control group, who received diagnostic training by reading a standard textbook, and an experimental group, who received diagnostic training by a combination of reading a standard textbook and AR-based training. Participants then encountered a tension pneumothorax scenario on the AR patient and were asked to list the symptoms they observed before making a final diagnosis. We recorded both the total amount of time participants required to make a final diagnosis as well as the number of correctly identified symptoms, and the results of the control and experimental groups were compared with a t-test. We found no significant difference between groups in the time needed to make a diagnosis. However, the AR-trained participants correctly identified a significantly greater number of symptoms (average: 12 symptoms) compared to the textbook-trained participants (average: 5 symptoms). These results indicate the potential of AR-based training to enhance detection and identification of cues in medical diagnostics.
  • Item
    Comparative analysis of Salmonella enterica serovar Typhi isolates from acute and chronic infections.
    (The Ohio State University, 2018-05) Eichar, Bradley; Gunn, John
    Salmonella enterica serovar Typhi (S. Typhi) is the causative agent of typhoid fever- a human-specific disease that results in approximately 200,000 deaths each year. With appropriate treatment, most patients recover from the acute stage of the disease. However, 3-5% of S. Typhi infections lead to a chronic infection and asymptomatic colonization in the host gallbladder – allowing carriers to unknowingly infect others despite no outward signs of illness. While it is known that S. Typhi mediates carriage through the formation of biofilms on the surface of cholesterol gallstones in the gallbladder, molecular mechanisms for chronic S. Typhi carriage are not well understood. This project compared genetic, molecular, and functional characteristics of 22 different S. Typhi isolates from confirmed acute and chronic infections, hypothesizing that the components necessary to mediate chronic carriage in the gallbladder may vary and be unique from acute clinical isolates. Biofilms of the isolates were examined utilizing a crystal violet quantification assay and confocal microscopy. The presence and relative abundance of Vi-antigen and lipopolysaccharide (LPS) were confirmed using dot blot assays with specific anti-Vi-antigen and anti-LPS antibodies and subsequent densitometric analysis. Additionally, immunofluorescent microscopy was used to evaluate Vi-antigen and LPS expression. The presence of cellulose was detected quantitatively via a microtiter-based calcofluor binding assay. Finally, the genome-sequence as well as pulsed-field gel electrophoresis (PFGE) patterns of all strains were determined. PFGE and the sequencing data informed phylogenetic relatedness determinations of the strains. Biofilm and extracellular matrix (cellulose, Vi-antigen, and LPS) expression studies revealed unexpected high variability between the S. Typhi strains. Preliminary analyses do not support the uniqueness of the chronic versus acute isolates. Additional analysis will provide a better understanding of how salmonellae enter and persist in the chronic carrier state, which may lead to targeted drug therapies to alleviate the carrier state.
  • Item
    Kicking in infants with perinatal stroke: can we predict ability and disability?
    (The Ohio State University, 2016-08) Durbak, Emily; Tanner, Kelly; Heathcock, Jill
    Stroke in very young infants is the most common cause of cerebral palsy, a chronic disorder of motor function impairments with lifetime physical, social, and emotional consequences. While stroke is typically diagnosed within the first week of life, cerebral palsy is diagnosed much later, at 18-24 months of age. Spontaneous kicking is one of the few early movements that has been shown to be indicative of motor ability in young infants with motor disorders. This is a longitudinal trial of children 2.5-7.5 months describing spontaneous kick quantity and quality in infants with and without stroke and those who go on to have CP. Infants were tested in 2 conditions: with and without toys. N=29 infants completed this study (n=11 with perinatal stroke (PS); n=18 typically developing (TD)). Medical chart review at 3 years of age determined cerebral palsy status of infants with stroke. Infants were videotaped kicking freely for eight, thirty-second trials, six with toys, two without. Videos were then analyzed by two trained blinded raters with overall agreement of 0.96. Infants with cerebral palsy showed a statistical interaction with infants without cerebral palsy for total kicks (p=0.019) and two of three kick subtypes (p=0.041; p=0.029). The no toy condition yielded more kicks than the toy condition for total kicks (p=0.025) and two of three kick subtypes (p=0.050; p=0.065). Our findings support the possibility that spontaneous kicking may be used to identify infants with motor deficits at much younger ages than is currently possible. The use of toys in lower extremity analyses is cautioned.
  • Item
    Elucidating the Role of UbE2K in the Post-Translational Regulation of the Tumor Suppressor PTEN
    (The Ohio State University, 2016-05) Lordo, Matthew; Majumder, Sarmila; Leone, Gustavo
    The tumor suppressor phosphatase and tensin homolog deleted in chromosome ten (PTEN) is one of the most frequently altered genes in human cancer. Germline mutations in PTEN result in autosomal dominant hamartomatous syndromes, including Cowden Syndrome, a disease that carries an increased risk for breast, thyroid, and endometrial cancer. Although genetic and epigenetic alterations account for PTEN loss in many cancers, in breast cancer there is a very low incidence of mutation in PTEN. Loss of PTEN has been shown to significantly increase the rate of tumor progression in multiple mouse models of breast cancer, including ErbB2 induced mammary tumorigenesis, and plays an important role in both the epithelial and stromal compartments. These data also correlate with human disease progression, as our studies demonstrate that patients with low PTEN protein levels have worse disease outcome compared to patients with high levels of PTEN protein (unpublished data). However, PTEN mRNA levels in this patient cohort are not predictive of disease outcome. Our observation that 30-40% of the breast cancer patients have a reduction in PTEN protein levels without significant alteration of its expression suggests that post-translational events plays a key role in regulating PTEN protein level in breast cancer. Thus, a global shRNA screen to identify putative regulators of PTEN protein was undertaken in the laboratory. A subset of the identified PTEN degradome are components of the ubiquitin-proteasome pathway. Treatment of cells with the proteasome inhibitor MG132 results in an increase in total PTEN protein, supporting our observation. We chose to study one of these genes, ubiquitin conjugating enzyme E2K (UbE2K), to elucidate its mechanism of action in regulating PTEN protein levels.
  • Item
    Exploring the Role of an Adipocyte-Specific Knockout of MHCII on Inflammatory-Driven Complications of Obesity in Female Pro-atherogenic Mice
    (The Ohio State University, 2021-05) Middendorf, Dana; Hsueh, Willa
    The adipocyte, in addition to functioning as an energy storage cell, is an important immune cell that links the innate and adaptive immune system. In obese humans, our laboratory has previously demonstrated that the major histocompatibility complex II (MHCII) pathway is one of the most differentially expressed between lean and obese adipocytes. Further work in high fat diet fed mouse models demonstrates that increased adipocyte MHCII expression resulted in pro-inflammatory T cell differentiation leading to insulin resistance and decreased glucose tolerance. In contrast, loss of adipocyte MHCII expression in aMHCII knockout male mice resulted in less inflammatory adipose tissue characterized by increased adipose tissue immunosuppressive regulatory T cells (Tregs) and prevented the development of insulin resistance and glucose intolerance induced by Western high fat diet (WD). Moreover, these mice had less atherosclerosis and less liver steatosis and inflammation, common complications of obesity. We, thus, aimed to understand the impact of the loss of aMHCII expression in female mice on these and other inflammatory complications. We crossed aMHCII KO mice with proatherogenic low density lipoprotein receptor knockout (LDLR-/-) mice, aged the female animals for one year to accelerate atherosclerosis and hepatic injury, and fed them WD for twelve weeks (n=12 control, n=12 aMHCII KO). Glucose (GTT) and insulin (ITT) tolerance tests, body fat composition and fasting blood were collected prior to WD start and at study end. At the time of sacrifice, visceral adipose tissue was collected for T cell flow analysis and gene expression, liver was collected for liver fat analysis and histology, and aortas were collected for atherosclerotic lesion area measurement. Results showed that the female aMHCII KO mice experienced a significantly lower percent weight gain over 12 weeks than wild-type (control) littermate control mice, despite demonstrating no significant difference in overall percent body fat. The knockout mice additionally demonstrated improved glucose tolerance after 12 weeks of WD, compared to the control mice (AUC control = 23090.0 +/- 520.9 vs. KO= 20442.3 +/- 657.9, P=0.0045), although no difference was observed in insulin sensitivity. No difference was found in circulating plasma triglyceride or cholesterol levels; however, liver fat accumulation was significantly decreased in KO mice (control= 17.14 +/- 0.93 %fat vs. KO= 13.89 +/- 1.04 %fat, P=0.039). Finally, no difference was found in atherosclerosis % lesion area of the aorta. These results support that the adipocyte knockout of MHCII leads to improvement in the metabolic and hepatic phenotype in female mice after 12 weeks on WD. However, as the females demonstrated some variation from the effects seen in male mice, further investigation into the cause of these differences is warranted.
  • Item
    Characteristics of Ohio women that did not receive reproductive and sexual healthcare
    (The Ohio State University, 2021-05) Foster, Abigail; Norris Turner, Abigail
    Objective: The American College of Obstetricians and Gynecologists (ACOG) recommends that a person with female reproductive anatomy receive their first Obstetrics and Gynecology (OBGYN) visit at ages 13-17, and continue to have annual women's health visits throughout their lifetime.1 Though this recommendation is well known, whether there is equitable access for all adult Ohioans with female reproductive anatomy is unknown. Previous research has determined that barriers to accessing reproductive healthcare in the United States are disproportionately experienced by marginalized individuals2 and it is critical to determine if this relationship holds true in Ohio, a state with vastly different community spaces from urban, to suburban neighborhoods and rural farming communities. Through this analysis, I analyze the characteristics of Ohio women who experienced reduced accessibility of reproductive and sexual healthcare. Methods: The Ohio Survey of Women is a population-representative survey that captures data about reproductive health and demographics, including age, race and sexuality, among women in Ohio. This survey was conducted by NORC in 2018 through 2019 and sampled among women of reproductive age living in Ohio (18-44 years). Using unadjusted logistic regression, I identified characteristics of Ohio women that are associated with not receiving an annual women's health visit in the past year. Results: A total of 2,613 participants completed the survey. After removing respondents who were missing data on the primary variables of interest, I was left with an analytic sample of 2,434. Race (Black OR= 1.18, CI= 0.75, 1.86; Hispanic OR= 0.91, CI= 0.49, 1,66; Multiracial/other OR=1.44, CI= 0.95, 2.16), living in rural Appalachia (OR=1.05, CI= 0.82, 1.32), and being currently pregnant (not pregnant OR= 1.52, CI= 0.77, 3.03) were not significantly correlated with not obtaining an annual women's visit. Individuals that identified as a sexual minority (OR=1.52, CI= 1.11, 2.08), were young (aged 18-24 OR=2.35, 95% CI= 1.86, 3.27), lacked health insurance at some point over the last year (OR= 4.51, CI= 3.10, 6.56), were not employed (OR=1.50, CI= 1.16, 1.94) and had low socioeconomic status (less than 75,000 U.S. dollars a year, some college or less OR=2.26, CI=1.72, 2.99; income more than 75,000 U.S. dollars, some college or less OR=1.86, CI= 1.28, 2.70; income less than $75,000, bachelor's degree or higher OR=1.43, CI=1.03, 1.99) had significantly higher odds of not obtaining an annual women's health visit in the last year. Conclusion: It is critical to identify which Ohio women are most likely to miss their annual women's health visit. When advocating for a system of nondiscriminatory healthcare access, we must identify the groups experiencing the most barriers to access in order to create interventions that better serve these groups.