Master of Science (MS), Wright State University, 2019, Microbiology and Immunology

Mouse models of eosinophil-associated diseases have been used to study the mechanisms of disease pathogenesis. In this study, mouse-derived bone marrow cells were used in long-term (6 and 9 months) cell cultures of differentiated eosinophils and macrophages. IL-5 was used to differentiate the stem cells to eosinophils and GM-CSF was used to propagate macrophages from the bone marrow stem cells. The maximum time period for observing the eosinophil cultures was 252 days which is censurably longer than the 18 days culture period observed by others. The results were assessed by describing the microscopic cell morphology by Wright staining, modified Giemsa staining and protein expression by immunofluorescent staining. The GMCSF-stimulated bone marrow cultures produced classically appearing monocyte/macrophages throughout the study and were used to compare the development of the eosinophils over the long-term period of observation. Differentiation of the BM cells was carried out using with growth factors (SCF, FLT3L) and cytokines (IL-5, GM-CSF) over the 252 days period. The most suitable culture plate for long-term of cell growth were thee 60 mm petri dishes. At 252 days, the eosinophils exhibited as bi-lobed nuclear shapes, comparable with human eosinophils. Dendrite-like ramifications were observed on the surfaces of these eosinophils. Long term culture of eosinophils in the presence of IL-5 contributed to formation of eosinophil extracellular traps (EETs) areas. Within the EETs the cells surface of eosinophils developed holes; the nucleus of such cells lost the “ring-like” or “lobular” morphology and appeared in de-condensed fashion. Within the EETs, the plasma membranes of eosinophils developed the protrusions containing cytoplasmic granules. The EETs and protrusions had not been observed previously in the mouse eosinophil models in vitro but were described in human eosinophils. Consequently, the long-term culture of mouse bone-marrow derived eosinophil (open full item for complete abstract)

Committee: Nancy J. Bigley Ph.D. (Advisor); Dawn P. Wooley Ph.D. (Committee Member); Marjorie Markopoulos Ph.D. (Committee Member) Subjects: Immunology; Microbiology

Master of Science, The Ohio State University, 2016, Dentistry

Objectives: Bone marrow mesenchymal stem cells (BM-MSCs) from limb bones have demonstrated promises in regenerating craniofacial bones; yet little is known about the potential of BM-MSCs from craniofacial bones. This study compared BM-MSCs isolated from limb and craniofacial bones in a commonly used preclinical large animal model, the pig. Methods: Bone marrow was aspirated from the tibia and mandible (symphysis) of 4-month-old pigs (n=4). Subsequently, BM-MSCs were isolated, expanded and confirmed by flow cytometry. To assess cell proliferation, cell doubling times were calculated from serial cell number counts over 2 weeks. Total mRNA was extracted from freshly isolated BM-MSCs and analyzed for gene expression using an Affymetrix GeneChip porcine genome array, followed by real-time RT-PCR for validation. Osteogenic capacity was assessed by quantifying alkaline phosphatase activity. Using temperature-responsive culturing plates, the abilities of BM-MSCs to form multi-layer cell sheets (for future in-vivo transplantation), along with cell viability and morphology, were evaluated by fluorescent labeling and histological staining. Results: BM-MSCs from both locations expressed MSC markers but not hematopoietic markers. Mandibular BM-MSCs proliferated significantly faster than tibial BM-MSCs (median cell doubling times: 2.25 vs. 2.80 days, Mann-Whitney U test, p<0.01). Without any osteogenic induction, mandibular BM-MSC alkaline phosphatase activities were 3.3-fold (factorial ANOVA, p<0.001) to those of tibial BM-MSCs. Microarray analysis on one hand confirmed that overall mandibular and tibial BM-MSC gene expressions are highly correlated with each other and genes related to osteogenesis and angiogenesis were strongly expressed, and on the other hand revealed that there were several dozen genes which were indeed differentially expressed between mandibular and tibial BM-MSCs. They include cranial neural crest-related genes nestin (1.23-fold) and BMP-4 (1.79-f (open full item for complete abstract)

Committee: Zongyang Sun DDS/PhD/MS/MSD (Advisor) Subjects: Dentistry

Doctor of Philosophy, The Ohio State University, 2015, Comparative and Veterinary Medicine

Cathepsin K (CatK), a cysteine protease, has been implicated in the process of bone resorption and inflammation. Selective inhibitors of CatK are promising therapeutic agents for the treatment of diseases associated with excessive bone loss and osseous inflammation, such as osteoarthritis, rheumatoid arthritis, periodontitis, osteoporosis, and multiple myeloma. Multiple reports have emerged over the last several years demonstrating the effect of different CatK inhibitors on osteo-inflammatory conditions. Therefore, the study of CatK inhibition as a target to prevent bone loss and inflammation, and influence bone marrow osseous progenitor cells, in a large animal model, is the subject of this dissertation. The horse was selected as the large animal model because this species suffers from ailments of adaptive bone remodeling in their sport performance and CatK inhibitors may serve as therapeutics in this species as well as serve as a large animal model for human applications. In the first phase of this work, we determined an optimal dose and dose interval for a CatK inhibitor (CatKI), VEL-0230, in healthy adult horses. Plasma pharmacokinetic (PK) and bone resorption biomarker [carboxy-terminal cross-linking telopeptide of type I collagen (CTX-1)] analyses were performed following single and multiple oral dose protocols of a CatKI (VEL-0230) in horses. Weekly administration of VEL-0230, at a dose of 4 mg/ kg body weight, provided effective inhibition of bone resorption in young exercising horses that returned to baseline within 7 days after drug withdrawal even after multiple doses. In the second phase of this work, we evaluated bone structure and turnover in healthy young exercising horses receiving repeated oral dosing of a CatKI in a randomized, controlled, double-blinded, prospective, sufficiently powered clinical trial. With the objectives of: 1. To determine whether repeated dosing of a CatKI produced a desired inhibition of the bone resorption biomarker (open full item for complete abstract)

Committee: Alicia Bertone PhD (Advisor); Maxey Wellman PhD (Committee Member); Prosper Boyaka PhD (Committee Member); Teresa Burns PhD (Committee Member) Subjects: Biology; Biomedical Research; Cellular Biology

Doctor of Engineering, Cleveland State University, 2010, Fenn College of Engineering

The ultimate goal of this project is to develop a biodegradable and implantable scaffold with precise surface topographies that can provide osteoconductive stimuli to connective tissue progenitor cells (CTPs), and subsequently, enhance bone regeneration applications without the complications of autogenous cancellous bone grafts. This dissertation presents the modification of surface microtextures to provide osteoconductive stimuli to CTPs for bone regeneration applications. First, the effect of surface topography on cell proliferation and osteogenic differentiation was validated through experiments using surface post microtextures and CTPs. Post microtextures accelerated CTP growth behaviors compared to smooth polydimethylsiloxane (PDMS) and standard cell culture dishes. Second, soft lithographic techniques were used to develop PDMS post microtextures with varying geometry and arrangement. 10 um diameter post microtextures with various inter-spaces (5, 10, 20, and 40 um) and post heights (5, 10, 20, and 40 um) were developed, and cultured with CTPs to establish optimal and precise surface post microtextures that can provide CTPs with an osteoconductive environment. Cells on post microtextures with 10 um height and 10 um inter-space exhibited higher cell number than other micro-posts with different heights or inter-spaces, and smooth surfaces. The results demonstrate a significant response of CTPs to topography, and suggest a practical role for optimal post size on textured materials in modifying CTP behavior. Third, substrate stiffness of various PDMS formulations was analyzed to investigate the effects on morphology, proliferation, and osteogenic differentiation of CTPs. Stiffer PDMS substrates with surface microtextures provided an enhanced osteoconductive microenvironment to CTPs relative to softer PDMS substrates. Finally, soft lithography techniques were successfully applied to biodegradable materials, including cellulose acetate (CA) and poly lactic-co-glycoli (open full item for complete abstract)

Committee: Shuvo Roy (Committee Chair); Joanne Belovich (Committee Co-Chair); Aaron Fleischman (Committee Member); Nolan Holland (Committee Member); Ronald Midura (Committee Member) Subjects: Polymers

Master of Science, The Ohio State University, 1961, Graduate School

Committee: Not Provided (Other) Subjects:

MS, University of Cincinnati, 2024, Medicine: Immunology

Bone marrow has the most mysterious anatomy of differentiation compared to the vast majority of other tissues. In fact, assessing blood cell production in situ is difficult due to a lack of strategies to examine differentiation, lineage commitment decisions, and tissue functionality with regard to spatial organizing cues. All blood cells originate from hematopoietic stem cells (HSCs). The bone marrow microenvironment contains HSCs and non-hematopoietic cells where they form a niche which directs hematopoietic output. At times of homeostasis or stress, microenvironmental cues direct HSCs to differentiate and proliferate and these hematopoietic niches are commonly referred to as HSC niches. It may be possible to develop novel therapeutic approaches for HSCs and their progeny, which are affected by microenvironment dysfunction. By using cold active protease generated from Bacillus licheniformis, we hope to understand the microenvironment through microdissection and sequencing techniques. Previous studies have shown that single cell RNA sequencing artifacts are significantly reduced compared to conventional methods [1]. Our method enables us to isolate vasculature cells from mouse sternums via microdissection, without using commonly used fixation steps for tissue imaging. Through this optimized technique, downstream analysis, such as scRNAseq of the dissected area can be performed to understand both the steady state as well as the diseased microenvironment of bone marrow.

Committee: Jonathan Katz Ph.D. (Committee Chair); Ian Lewkowich Ph.D. (Committee Member); Daniel Lucas Ph.D. (Committee Member) Subjects: Immunology

Master of Science, The Ohio State University, 2024, Veterinary Clinical Sciences

Equine superficial digital flexor tendon (SDFT) injuries heal primarily through fibrosis, in part due to persistent macrophage-induced inflammation throughout the healing process. Despite the low retention and short life of bone marrow mesenchymal stem cells (BM-MSC) following intratendinous injections, benefits including reducing local inflammation, improving histological structure, and reducing tendon re-injury rates, in experimental and naturally occurring equine tendonitis, are well-known. Extracellular vesicle (EV) release at the healing milieu is one of the speculated mechanisms for the immunomodulatory and anti-fibrotic benefits of BM-MSC. EVs are secreted membrane-bound cytokine, growth factor and mRNA/miRNA cargo reflective of the parent cells and facilitate intercellular communication. From a clinical standpoint, EVs circumvent donor site morbidity, time lag for culture expansion and allogenic cell-related immunogenicity limitations of BM-MSC and comprise an ‘off-the-shelf' cell-free therapy for equine tendon injuries.    This research investigates the mitogenic, anti-inflammatory, and extracellular matrix (ECM) remodeling effects of equine BM-MSC-derived EVs (BM-EV) on tenocytes cultured on acellular SDFT matrix. We hypothesized that (1) BM-EV increases the adherence and proliferation of tenocytes during culture on acellular SDFT matrix, and (2) BM-EV modulates inflammation and stimulates extracellular matrix (ECM) remodeling during direct (cell-to-cell contact) tenocyte-macrophage co-culture on acellular SDFT matrix.   Undifferentiated (7-day basal media), pro-inflammatory (6-day GM-CSF and 24-hour LPS+IFN-γ priming) and regulatory (6-day basal media and 24-hour IL-4+IL-10 priming) macrophages were generated from peripheral blood CD14+ monocytes. Passage 3 tenocytes (CD90+105+27-45-) were isolated from freshly euthanized cadaveric forelimb mid-metacarpal SDFT via collagenase I digestion and monolayer culture.   BM-EV were isolated via ultra (open full item for complete abstract)

Committee: Sushmitha Durgam (Advisor); Alison Gardner (Committee Member); Laura Hostnik (Committee Member); Amit Sharma (Committee Member) Subjects: Comparative; Medicine

PhD, University of Cincinnati, 2023, Medicine: Immunology

The bone marrow (BM) is a hematopoietic (blood forming) organ that dynamically adjusts output to accommodate demand. Hematopoietic stem and progenitor cells (HSPC) reside within the BM and sequentially differentiate to feed each major lineage. All lineages are derived from the hematopoietic stem cells (HSC). HSC give rise to erythroid progenitors that terminally differentiate into red blood cells (RBC); lymphoid progenitors that generate dendritic cells (DC), B and T-cells that provide long term adaptive immunity; and myeloid progenitors that give rise to granulocytes, monocytes and macrophages which coordinate the rapid innate immune responses. These progenitors inhabit a network of non-hematopoietic (stromal) cells that maintain and regulate progenitors. Different tissue cells comprise the stromal cell compartment, including endothelial cells that make up vessels, osteoblasts that make up bone, fat cells, nerve cells, and perivascular stromal cells which are derived from mesenchymal stem cells (MSC). Stromal cells provide essential retention and differentiation cues, whose function depends on the cellular source of these cytokines. Besides stromal cells, mature hematopoietic cells can also directly and indirectly regulate hematopoietic progenitors. Together, the combination of stromal and hematopoietic cells, along with their cytokine products, make up spatially distinct niches in the BM. Acute and chronic insults can lead to inflammation which dramatically changes BM output. The subsequent cytokine storm can regulate progenitors directly or indirectly via a stromal cell intermediary. While inflammatory responses are advantageous to pathogen clearance, in other cases, inflammation leads to reprogramming of progenitors and BM output. When inflammation is sensed by progenitors, they become activated, proliferate, mobilize, or terminally differentiate into mature hematopoietic cells. Inflammation has emerged as a major regulator of both non-malignant (open full item for complete abstract)

Committee: Daniel Lucas Ph.D. (Committee Chair); Damien Reynaud Ph.D. (Committee Member); Leah Claire Kottyan Ph.D. (Committee Member); David Hildeman Ph.D. (Committee Member) Subjects: Immunology

MS, University of Cincinnati, 2022, Medicine: Biomedical Research Technology

The majority of acute myeloid leukemia (AML) cases have chromosomal abnormalities, composed of translocations or aneuploidies. The oncogene DEK was first identified in acute myeloid leukemia as a translocation with NUP214. However, the role of full length DEK in AML is poorly understood. DEK does have a role in DNA repair pathways, including Homologous Recombination and Non-homologous End-Joining, where it promotes DNA-PK activity. This study investigates the development of a DNA-pk mutant and DEK-deficient mouse model. In this novel mouse model, tissue histology, along with blood and bone marrow immunophenotyping, determined that dual loss of DNA-pk and DEK generated a murine model of leukemia. DNA-pk mutant and DEK deficient mice exhibited a reduction in early hematopoietic stem and multipotent progenitor cells while also displaying an increase in the number of granulocyte-monocyte progenitors and differentiated granulocytes and neutrophils. Generation of a mouse model with dual loss of DEK and DNA-pk will serve as a tool to further investigate the role of full length DEK in leukemia.

Committee: Lisa Vinnedge Ph.D. (Committee Member); Jason Tchieu Ph.D. (Committee Member); Marie-Dominique Filippi Ph.D. (Committee Member) Subjects: Oncology

PhD, University of Cincinnati, 2021, Medicine: Molecular, Cellular and Biochemical Pharmacology

Atherosclerosis is a chronic inflammatory disease that results from the accumulation of cholesterol-containing LDL particles in the vessel wall, and is the underlying pathology for many cardiovascular diseases. Current clinical guidelines focus on LDL-cholesterol lowering as a means of cardiovascular disease prevention. The immune system also plays a major role in the pathogenesis of this disease, and emerging evidence points to the therapeutic potential of targeting inflammation to combat cardiovascular diseases. Studies have identified dual roles of apolipoprotein E (apoE) and apoE receptors in modulating plasma lipids and inflammation, and have implicated these proteins as key players in atherogenesis. Moreover, genome-wide association studies have linked polymorphisms in the genes encoding these proteins with altered cardiovascular disease risk in humans. ApoE was first identified as a component of circulating lipoproteins, and the importance for ligand-receptor interactions between lipoprotein-associated apoE and hepatic endocytic receptors for maintaining healthy cholesterol levels has been since well established. However, the roles of apoE and apoE receptors expressed in other tissues and their involvement in the pathogenesis of atherosclerosis have not been fully elucidated. The goals of this dissertation research are to (1) investigate the impact of human APOE polymorphisms on macrophage-driven mechanisms of atherosclerosis, and (2) evaluate the effect of global and immune-cell specific expression of a novel mutation of the apoE receptor, LRP1, on development of metabolic disease and atherosclerosis. The functions of macrophage expression of the different human apoE isoforms – apoE2, apoE3, and apoE4 – in modulating atherogenesis was assessed by transplantation of donor bone marrow expressing the apoE isoforms into recipient ApoE-/- mice. Despite the similar plasma cholesterol levels between groups, we observed significant reduction in atherosclerosis (open full item for complete abstract)

Committee: David Hui Ph.D. (Committee Chair); Guo-Chang Fan Ph.D. (Committee Member); Marie-Dominique Filippi Ph.D. (Committee Member); Evangelia Kranias Ph.D. (Committee Member); Alex Lentsch Ph.D. (Committee Member); William Miller Ph.D. (Committee Member) Subjects: Pathology

PhD, University of Cincinnati, 2021, Medicine: Cancer and Cell Biology

Hematopoietic stem cells (HSCs) regenerate the hematopoietic tissue during stress hematopoiesis, requiring that HSCs come out of quiescence and re-fill the depleted pool, as well as self-renew to maintain a healthy HSC pool. Injuries associated with stress hematopoiesis include myeloablative chemotherapy, infection/inflammation and allogeneic bone marrow transplantation. However, self-renewal and differentiation must be tightly controlled. Too much differentiation results in an exhaustion of the HSC pool and bone marrow failure, while too much self-renewal can lead to leukemogenesis. The transforming growth factor ß (TGFß) signaling pathway has been well-studied to regulate HSCs during steady state hematopoiesis. However, very little is known about its role in regulating HSCs during stress hematopoiesis. Moreover, its role in HSC regulation is both context-dependent and concentration dependent. Using a mouse model to conditionally overexpress an active form of the main TGFß ligand, aTGFß1, we found that increased aTGFß1 caused very little impact in the hematopoietic system during steady state hematopoiesis, driving a very mild neutropenia and bone marrow dysplasia during aging. However, aTGFß1 overexpression prevented the murine hematopoietic system from recovering to normal levels of peripheral blood leukocyte and platelet levels after chemotherapeutic stress using 5-fluorouracil. Moreover, during the recovery phase aTGFß1-overexpressing mice had increased HSC cell cycling and DNA double strand breaks compared to controls. Separately, acute inflammation-driven stress caused by polyinosinic:polycytidilic acid (pIC) drove pancytopenia, bone marrow dysplasia and splenomegaly, but expanded the hematopoietic stem and progenitor (HSPC) pool long after the short-term effects of pIC had resolved, phenotypes similar to the human bone marrow failure myelodysplasia. Mechanistically, we found that aTGFß1 overexpression altered the HSC transcriptome after pIC-driven str (open full item for complete abstract)

Committee: Marie-Dominique Filippi Ph.D. (Committee Chair); Paul Andreassen Ph.D. (Committee Member); Jose Cancelas-Perez M.D. (Committee Member); H. Leighton Grimes Ph.D. (Committee Member); Gang Huang Ph.D. (Committee Member); Damien Reynaud (Committee Member) Subjects: Health Sciences

Doctor of Psychology (Psy.D.), Xavier University, 2020, Psychology

Trait mindfulness is stable over time and has been shown to predict health-related quality of life (HRQOL), mood disturbances, and other symptoms of distress in certain cancer populations. In the current study of 43 bone and blood cancer patients receiving a bone marrow transplant (BMT), the researcher investigated whether levels of trait mindfulness served as a protective factor from the psychological and physical burden of the procedure. Additionally, the association between two commonly used mindfulness measures was examined. The researcher utilized two baseline measures of trait mindfulness along with three measures of treatment-related outcomes—HRQOL, mood disturbances, and post-traumatic stress disorder (PTSD) symptoms—both before and after BMT. Correlational analyses assessed the degree of association between the two mindfulness measures. Six hierarchical regression analyses tested the predictive power of trait mindfulness on the three outcomes. Contrary to the hypotheses, results indicated that levels of trait mindfulness failed to significantly predict ratings of HRQOL, mood disturbances, and PTSD symptoms following BMT. This BMT-sample was unique in its comparatively high levels of trait mindfulness. Two measures of trait mindfulness, the FFMQ-15 and MAAS, were moderately, positively correlated, but only the FFMQ-15 Nonreacting subscale was significantly correlated with the MAAS. Our results apply to oncological settings, suggesting BMT patients may benefit from interventions that teach how to utilize mindfulness for healthier coping. Future research should consider measuring the effects of cancer treatment on mindfulness levels, as well as the associations between mindfulness and treatment-related outcomes up to a year post-BMT.

Committee: Jennifer Gibson Ph.D. (Committee Chair); Lyn Sontag Psy.D., ABPP (Committee Member); Nicholas Salsman Ph.D., ABPP (Committee Member) Subjects: Psychology; Psychotherapy

PhD, University of Cincinnati, 2020, Medicine: Pathobiology and Molecular Medicine

Over the last few decades, many studies have clarified and highlighted the importance of bone marrow niche components in both normal and aberrant hematopoiesis. A mutation in the niche was often sufficient to trigger the onset of a blood abnormality. The bone marrow vascular niche has been shown to be of vital importance in the maintenance of hematopoietic stem cells (HSCs). Bone marrow endothelial cells are major components of the vascular niche and have an intimate and dynamic relationship with HSCs. To determine how a mutation in endothelial cells can disturb the microenvironmental interactions and their functional consequences, we used a tamoxifen-inducible Tie2-CreERT2;tdTomato;KrasG12D mouse model to activate oncogenic Kras in endothelial cells. Complete blood counts and flow cytometry data revealed that the KrasG12D mice had increased leukocytosis and an increased myeloid cell percentage with concurrent decrease in lymphocytes. The mice were euthanized and their bones and spleens were harvested. Surprisingly, flow cytometry analysis revealed no major changes in the bone marrow progenitor compartments. On the other hand, the spleens of the KrasG12D mice were bigger and analysis of the progenitor compartments showed an increase in the short-term HSCs, suggesting that the spleen also contributed to the observed phenotypes. Colony-forming units performed from the bone marrow and spleen cells of KrasG12D mice showed a greater colony-forming potential. However, competitive transplant studies revealed an impairment of the long-term functionality of the HSCs in KrasG12D mice, as evidenced by decreased donor chimerism in all blood lineages analyzed. In addition, imaging studies showed that HSCs are located further away from the blood vessels of mutant mice. To obtain some mechanistic insight of the pathways involved, CD31+ tdTomato+ endothelial cells were sorted and RNA sequencing was performed. Subsequent analyses uncovered an up-regulation in the inflammatory an (open full item for complete abstract)

Committee: Yi Zheng Ph.D. (Committee Chair); Jose Cancelas-Perez M.D. Ph.D. (Committee Member); Marie-Dominique Filippi Ph.D. (Committee Member); Daniel Starczynowski Ph.D. (Committee Member); Meifeng Xu M.D. Ph.D. (Committee Member) Subjects: Molecular Biology

Doctor of Philosophy, The Ohio State University, 2019, Neuroscience Graduate Studies Program

Spinal cord injury (SCI) is a devastating condition causing severe loss of motor, sensory, and autonomic function below where the injury occurs. injury the cord causes severe damage to cell components, blood-spinal cord barrier, and axon tracts. However, SCI also causes hemorrhage and the release of cell content that initiate a secondary wave of inflammation and additional tissue damage. Therefore, targeting the immune system acutely after injury may prevent further damage to the cord. However, components of the neuroinflammatory response are also beneficial for spinal cord repair long-term. A deeper understanding of these divergent aspects of post-SCI inflammation and immune function could aid in the development of therapies that modulate immune responses to be less damaging while promoting their reparative properties. Translating promising therapeutics from bench to bedside has been historically grim in the context of central nervous system (CNS) injury and disease. Within the context of neuroimmunology, there are many differences in the immune systems of rodents and humans that may preclude the translation of therapeutic strategies, warranting a further understanding of the human immune response to SCI. However, studies involving human or large animal models are prohibitively expensive, difficult to perform, and preclude in vivo mechanistic studies. Novel models of human immune function within the context of CNS injury and disease are warranted. Humanized mice are immunocompromised mice engrafted with human immune systems, creating an in vivo model of human immune function. The premise of humanized mice originally came from a need to model HIV infection. Humanized mice have since allowed for the investigation of human immunobiology in the context of hematopoiesis, immunotherapy, immune-oncology, and infectious disease. Humanized mice represent a promising tool to model human immune responses to CNS injury and disease, including SCI. The first section of thi (open full item for complete abstract)

Committee: Phillip Popovich Ph.D. (Advisor); Jonathan Godbout Ph.D. (Committee Member); Andrea Tedeschi Ph.D. (Committee Member); Adrienne Dorrance Ph.D. (Committee Member); Stefan Niewiesk Ph.D., D.V.M. (Committee Member) Subjects: Biomedical Research; Immunology; Neurobiology; Neurosciences

Doctor of Philosophy, The Ohio State University, 2019, Pharmacy

HLA-matched unrelated donor (MUD) hematopoietic stem cell transplantation (HSCT) is used as a potential curative therapy for otherwise fatal hematological cancers, including but not limited to acute and chronic leukemia, Hodgkin and non-Hodgkin lymphoma, and myelodysplastic syndrome (MDS). MUD-HSCT involves the transfer of healthy individual (donor) bone marrow stem cells to an unhealthy patient (recipient). Despite improvements in human leukocyte antigen (HLA) matching algorithms, treatment protocols and patient care overall survival (OS) rates profoundly decrease in the first 1-year post-transplant. The largest predictor of mortality during this time frame is HLA matching, emphasizing the importance of genetic variation in determining survival. Despite the clear importance of genetics to survival following transplant, the relationship of non-HLA genetic variation remains underexplored. In this dissertation, we characterized and explored the independent and joint non-HLA genetic contributions to survival outcomes in donors and recipients with acute leukemias or MDS using pre-existing genome-wide association study (GWAS), Determining the Influence of Susceptibility COnveying Variants Related to one-Year mortality after BMT (DISCOVeRY-BMT), a study of 3,532 donor-recipient pairs with survival, clinical and genomic data available. We used DISCOVeRY-BMT to replicate or validate all previous literature that investigated survival outcomes after allogeneic HSCT. None of the previous literature was reproduced, likely due to the previous cohorts comprising heterogeneous samples (testing with multiple diseases) or small sample sizes. Next we developed an R/Bioconductor software package called gwasurvivr. This package takes input from popular genomics software and performs multivariate Cox regression survival analysis in a scalable and efficient manner. We integrated gwasurvivr into an automated pipeline that performs full survival GWAS analyses and conducts meta-analysis. (open full item for complete abstract)

Committee: Lara Sucheston-Campbell PhD (Advisor); Guy Brock PhD (Committee Member); Moray Campbell PhD (Committee Member); Shili Lin PhD (Committee Member) Subjects: Bioinformatics; Biostatistics; Epidemiology; Genetics; Pharmaceuticals; Statistics

Doctor of Engineering, Cleveland State University, 2018, Washkewicz College of Engineering

The intrinsic repair mechanism of human heart is not sufficient to overcome the impact placed by adverse pathological conditions, such as myocardial infarction (MI). Current clinical approaches have played significant role in reducing the mortality rate; however, these approaches do not replace the lost cells and tissues of the myocardium. Human bone marrow-derived mesenchymal stem cells (BM-MSCs) are gaining attention in cardiac therapy due to their ability to differentiate into cardiomyocyte like cells and release a wide repertoire of paracrine factors. However, clinical trials of longer duration have shown mixed results in improving cardiac functions. In addition, in-depth studies on the secretome profile of MSCs/ differentiated MSCs and optimal approaches to modulate their outcomes are still lacking. Hence, in the first project of this study, we investigated the role of cell-cell interactions (MSC spheroids), cell-matrix interactions (collagen concentration, topography) and cell-signaling cues [5-azacytidine (aza)] on the cardiac differentiation of BM-MSCs. We found that collagen hydrogel (2 mg/ml), in the presence of 10 µM of aza, offered higher cell survival and caused time-dependent cardiomyogenic evolution of MSC spheroids. We also identified that canonical Wnt/ß-catenin signaling pathway primarily mediated the observed benefits of aza on cardiac differentiation of MSC spheroids. In the second project, we quantified the secretome profile and matrix synthesis by collagen gel-laden BM-MSC spheroids, under optimized culture conditions from the first project, and extended our investigation to MSC spheroids within human fibroblasts-derived collagen. Human collagen promoted higher matrix synthesis over time but severely impacted cell proliferation. The release of inflammatory cytokines was drastically reduced with spheroid formation and collagen cultures, specifically within human collagen. In the last project, we examined the influence of adult human cardiomyocyt (open full item for complete abstract)

Committee: Chandra Kothapalli (Committee Chair); Moo-Yeal Lee (Committee Member); Nolan B. Holland (Committee Member); Mekki Bayachou (Committee Member); Anand Ramamurthi (Committee Member) Subjects: Biomedical Engineering

Doctor of Philosophy, The Ohio State University, 2018, Comparative and Veterinary Medicine

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a life-saving therapy both for malignant and non-malignant diseases. The success of allo-HSCTs, however, is limited by acute graft-versus-host disease (aGVHD), a frequent complication that remains a leading causes of non-relapse mortality following allo-HSCT. The pathogenesis of aGVHD involves donor T cells which target human leukocyte antigen mismatched host tissues, causing tissue injury through pro-inflammatory cytokine secretion and direct cytotoxicity. The morbidity and mortality associated with aGVHD pose a major barrier against the wider application of allo-HSCT as a curative modality. Thus, better understanding of aGVHD pathogenesis and novel therapeutics are needed. Modulation of T cell function, broadly, depends on control of gene expression. Two well-studied modes of modulating gene expression are noncoding RNAs and epigenetic modifications. Using unbiased approaches, we identified multiple microRNAs that are upregulated during aGVHD. We validated two of these, T-cell intrinsic miR-155 and serum miR-29a, due to their pivotal role in regulating the adaptive immune system. First, we investigate the molecular mechanisms by which miR-155 modulates T cell function in aGVHD. We identify that miR-155 expression in both donor CD8+ T cells and conventional CD4+ CD25- T cells is pivotal for aGVHD pathogenesis. Furthermore, we show that miR-155 strongly impacts alloreactive T cell expansion through proliferation and exhaustion as well as function by promoting a pro-inflammatory Th1 phenotype. Finally, we demonstrate that miR-155 expression in donor T cells regulates chemokine-dependent migration and infiltration into target organs. These findings provide novel insight into the role of miR-155 in regulating T cell function post-transplant and are convincing biological rationale to justify investigation of novel antagomiR-155 therapeutics to prevent or minimize aGVHD. Next, we strive to identify s (open full item for complete abstract)

Committee: Ramiro Garzon MD (Advisor); Michael Caligiuri MD (Committee Member); Renukaradhya Gourapura DVM, MS, PhD (Committee Member); M. Judith Radin DVM, PhD (Committee Member) Subjects: Immunology; Molecular Biology; Oncology

Doctor of Nursing Practice Degree Program in Population Health Leadership DNP, Xavier University, 2018, Nursing

The effects of sleep loss are a population health issue for the 65.7 million family caregivers in the United States (U.S.). Between 60-95% of caregivers report poor sleep quality because of sleep disruption. The purpose of this Doctorate of Nursing Practice (DNP) scholarly project was to evaluate the effectiveness of a combination of interventions, referred to as a sleep bundle, to improve sleep quality for caregivers of children with chronic conditions such as bone marrow transplant (BMT) using the FADE (Focus Analyze Develop Execute Evaluate) quality improvement (QI) methodology. It was hypothesized that the sleep bundle will decrease sleep disruption, and ultimately improve the sleep quality of primary caregivers of BMT patients. Specific aims to test the hypothesis included: (1) Compare sleep efficiency and quality in caregivers with and without the sleep bundle; (2) Identify and mitigate barriers and obstacles to implement the sleep bundle. Although the sleep bundle was followed reliably the nights of sleep bundle intervention phase; the sleep bundle was not found to be statistically significant in improving the sleep efficiency and sleep quality of caregivers of BMT patients. Additional evaluation of the sleep bundle and its impact on improving the sleep efficiency and quality of caregivers is needed with a larger sample size. Possible alteration of specifics interventions combined in the sleep bundle should also be explored.

Committee: Elizabeth Bragg PhD, RN (Committee Chair); Robin Saxon DNP, RN (Committee Co-Chair) Subjects: Health Care; Health Sciences; Medicine; Nursing; Psychology

Doctor of Psychology (Psy.D.), Xavier University, 2014, Psychology

High levels of distress are common among cancer patients, although few are referred for professional psychosocial care. Leading institutions such as the NCCN strongly recommend routine assessment of distress. There were three primary aims for this study: To establish a trajectory of distress for Bone Marrow Transplant inpatients throughout hospitalization; to determine variables that contribute to patient distress; and, to validate a new screening measure for distress specifically developed for use with BMT inpatients. Eighty-five BMT inpatients participated. No point in the trajectory of hospitalization could be identified as the highest point of distress. The point of lowest distress was at the time of discharge. As hypothesized, emotional, practical, family, physical, perceived level of personal control, meaning of life event, religious coping style, and perceived level of social support significantly contributed to patient distress throughout hospitalization. The Jewish Hospital Bone Marrow Transplant Unit Distress Screening Measure was validated as a screening measure for use with BMT inpatients. The ability to find meaning in life, maintain a positive religious coping style, a perception of personal control, and the support of others were identified as potential protective factors for patients undergoing BMT. Specifically assessing for these factors prior to admission and throughout hospitalization can provide clinicians with a more comprehensive understanding of patient strengths and limitations and help to identify patients who may benefit from additional support throughout BMT.

Committee: Renee' Zucchero Ph.D. (Committee Chair) Subjects: Psychology

Doctor of Philosophy, The Ohio State University, 1985, Graduate School

Committee: Not Provided (Other) Subjects: Biology