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

BRCA1 pathogenic variants are well-established risk factors for hereditary breast and ovarian cancer (HBOC). The increasing accessibility and affordability of genetic testing have led to the identification of individuals with BRCA1 variants of uncertain significance (VUS), creating a clinical challenge due to the rarity of these variants. This issue disproportionately affects women from minority backgrounds, who often face a high prevalence of VUS in their genetic screening results. The American College of Medical Geneticists (ACMG) guidelines stipulate that clinical recommendations cannot be made until these VUS are reclassified, leaving many individuals stranded in the healthcare system without access to advancements in preventive medicine. To address this gap, ACMG has permitted the use of functional assays to experimentally assess the effects of BRCA1 variants. This dissertation aims to identify the genetic and environmental factors contributing to the gaps in care for minority women dealing with HBOC, redesign the analysis pipeline for multiplexed DNA repair assays, introduce a novel multiplexed repair assay based on cisplatin resistance (CR), and adapt this pipeline to functional limited datasets. Our research successfully functionally characterized 2271 variants from the carboxy terminus for homology-directed repair function (HDR) and 1427 variants for CR. Notably, we observed consistent results between the two multiplexed functional assays, especially for non-functional variants located within critical regions of the BRCA1 protein essential for tumor suppression. Variants classified in the multiplex HDR assay exhibited 81% specificity and 93% sensitivity, while those in the multiplex CR assay demonstrated 100% specificity and 86% sensitivity. Furthermore, the functional categorizations of the variants correlated with known clinical significance and other BRCA1 functional assays. Utilizing the redesigned analysis pipeline, we reanalyzed previously publishe (open full item for complete abstract)

Committee: Jeffrey Parvin (Advisor); Amanda Toland (Committee Member); Tasleem Padamsee (Committee Member); Michael Freitas (Committee Member) Subjects: Biomedical Research; Cellular Biology; Health; Health Sciences; Information Science; Medicine; Public Health

Doctor of Philosophy, The Ohio State University, 2022, Chemical Engineering

For many people with acute organ failure or genetic conditions such as cystic fibrosis, organ transplantation is the only treatment option. The number of patients on the organ transplant waiting list is over 100,000 with thousands of people on the list dying each year while awaiting a life-saving organ transplant. Traditional organ storage is done using static cold storage (SCS) whereby organ allografts are kept in hypothermic conditions to decrease metabolism and slow tissue death. The harsh ex vivo preservation conditions of SCS mandate a high standard for graft utilization with marginal organs often being discarded for fear of causing graft dysfunction upon transplantation. Normothermic machine perfusion (NMP) represents a method of ex vivo organ preservation to reduce ischemic time and hypothermic organ injury such that grafts are better supported before utilization. NMP has been shown previously to also be able to resuscitate marginal organs that would have otherwise been discarded to be viable for transplantation. A current shortcoming of NMP is the steep oxygen (O2) demand of cellular respiration at normothermia and the metabolic debt acquired by organs during ischemic time post-mortem. To ameliorate this problem, O2 carriers have been used in perfusates; however, there is not yet an optimal O2 carrier for NMP. Red blood cells (RBCs) are prone to hemolysis in ex vivo perfusion and previous commercially available RBC substitutes have had large quantities of cytotoxic low molecular weight (LMW) hemoglobin (Hb) species (<500 kDa). This work describes the scale-up of a next generation polymerized human Hb (PolyhHb) as an RBC substitute in NMP. Previous work in this lab has synthesized a safer PolyhHb with LMW species purified out of solution but had only been made at the bench-top scale. Successfully increasing the process scale to the pilot scale makes this PolyhHb feasible for use in large animal NMP and eventually in clinical trial applications. The (open full item for complete abstract)

Committee: Andre Palmer (Advisor); Bryan Whitson (Committee Member); Aleksander Skardal (Other); Jeffrey Chalmers (Committee Member); Sylvester Black (Committee Member) Subjects: Chemical Engineering

Ph.D., Antioch University, 2019, Leadership and Change

Scientists debunked the belief that breast cancer is always viral with the mid-90s discovery of the first hereditary genetic mutation linked to a significantly higher-than average chance of breast and ovarian cancer. This genetic condition, called Hereditary Breast and Ovarian Cancer (HBOC), passes the mutation from generation to generation in a family. Thousands of variations of such mutations exist, and carriers account for 10 to 15% of all breast cancer, and up to 20% of ovarian (Childers et al., 2017). In addition, genetic testing uncovered a rapidly rising number of healthy people (never had breast/ovarian cancer) who are also carriers, flooding healthcare providers seeking potential options to reduce their elevated risk. Those prophylactic measures are invasive, permanent and can cause physical—and emotional—scarring. As a newer medical phenomenon, few, if any, studies address the potential psychological implications, which include fear, anxiety, guilt, family tension, and more. Using narrative inquiry methodology, this study analyzes the authentic lived or felt experiences of individuals when they learn that they have inherited a mutation that significantly increases their risk of breast, ovarian and related cancers, and their choices that directly affect their effort to outrun a cancer that may never come. This dissertation is accompanied by the author's MP4 video introduction and is available in open access at AURA: Antioch University Repository and Archive, http://aura.antioch.edu/ and Ohiolink ETD Center, https://etd.ohiolink.edu/

Committee: Elizabeth Holloway PhD (Committee Chair); Jon Wergin PhD (Committee Member); Piri Welcsh PhD (Committee Member) Subjects: Alternative Medicine; Behavioral Psychology; Communication; Educational Leadership; Families and Family Life; Genetics; Health; Health Care; Health Care Management; Health Education; Journalism; Psychology

Master of Science, The Ohio State University, 2019, Genetic Counseling

Although the clinical importance of cascade testing in families with hereditary cancer syndromes is well documented, complicated discussions can arise when genetic information is shared and subsequent discussions among relatives are often complicated. Novel communication aids should be considered to assist aid in these conversations. To investigate one possible method, we evaluated the theoretical impact of receiving unsolicited information about genetic testing performed in one's family through a video that could be shared via text or social media. Participants (N=399) viewed a video describing a relative's recent BRCA+ diagnosis and the potential impact on themselves. They also completed a survey with questions regarding thoughts on the message, hypothetical willingness to act on results, and other measured variables. The electronic survey instrument was built using the Health Belief Model, which postulates that an individual is more likely to engage in a behavior if they perceive greater risk in their severity and susceptibility, greater benefits than barriers to engagement in the behavior, self-efficacy, and a cue to action. This framework was used to measure participants' willingness to undergo genetic testing, seek out a genetic counselor, discuss genetic testing with their doctor, and talk to family members about their family history of cancer. Characteristics shown to impact intent to take action in these categories included intolerance of uncertainty, having a close family history of cancer, and greater family dynamics (ps < .05). A majority of participants (75.95%, N=300) would undergo genetic testing if it cost $100 or less. Additionally, a majority of participants (70%, N=281) were willing to meet with a healthcare professional to discuss genetic testing. Understanding potential reactions to receiving unsolicited genetic information is the first step in investigating novel communication aids in cascade testing.

Committee: Leigha Senter-Jamieson MS, LGC (Advisor); Kristen Carpenter PhD (Committee Member); Lindsey Byrne MS, LGC (Committee Member) Subjects: Genetics

Doctor of Philosophy, The Ohio State University, 2017, Chemical Engineering

In the United States, millions of people are affected by chronic wounds every year. Chronic wounds are characterized by low levels of oxygen (O2) and high levels of pro-inflammatory macrophages that fail to transition to the anti-inflammatory, pro-healing macrophages. There is a need for a therapeutic that simultaneously targets multiple deficiencies of chronic wounds in order to promote healing and avoid devastating outcomes. The main goal of this dissertation is to engineer the oxygen affinity of pure tense state (T-state) and relaxed state (R-state) polymerized hemoglobin (Hb) (PolyHb) mixtures. These mixtures could be topically administered to a chronic wound in order to accelerate the wound healing process by increasing the oxygen flux and exerting additional therapeutic effects by attracting and promoting pro-healing, anti-inflammatory macrophages. To our knowledge, none of the Hb wound healing therapeutics currently in existence use PolyHb mixtures. The partial pressure of oxygen (pO2) of a chronic wound can range from 2-20 mm Hg. By controlling the oxygen affinity (P50 ) of the PolyHb mixture, it is possible to control the pO2 range where the majority of oxygen delivery occurs at the wound site. Therefore, engineering PolyHb mixtures possessing a P50 range of 2-30 mmHg should elucidate the role P50 plays in accelerating wound healing and increasing wound pO2. This dissertation focuses on the development, characterization, and oxygenation potential of human and bovine PolyHb mixtures consisting of pure R-state and T-state PolyHb. This work presents analysis of the biophysical properties of R-state and T-state mixtures for both human and bovine polymerized Hb (PolyhHb and PolybHb) (Chapter 2). In addition, presented is the oxygenation potential of PolyHb mixtures using Comsol Multiphysics to simulate O2 transport in a hepatic hollow fiber bioreactor (Chapter 3). We produced PolyHb by polymerizing Hb using the chemical crosslinker glutar (open full item for complete abstract)

Committee: Andre Palmer Dr. (Advisor); David Wood Dr. (Committee Member); Jeffery Chalmers Dr. (Committee Member) Subjects: Chemical Engineering

Master of Science, The Ohio State University, 2016, Genetic Counseling

Introduction: Hereditary breast and ovarian cancer syndrome (HBOC) is a cancer-predisposition syndrome that affects both men and women, with more significant cancer risk elevations in women. Because there are well-established guidelines for cancer risk reduction and prevention in HBOC, it is critical that health care professionals understand information-sharing patterns among patients to facilitate communication processes and identify at-risk family members. Dissemination of familial genetic risk information in females with HBOC is well defined, but knowledge about how males share this information is limited. The aims of this study include: to describe participants' feelings and opinions about HBOC; to ascertain participants' extent of information sharing with family and medical personnel; and to describe the needs of participants for information and resources provided by genetic counselors and other health care providers. Methods: We interviewed 21 primarily Ashkenazi Jewish men who were accrued through Facing Our Risk of Cancer Empowered (FORCE). Interviews focused on family cancer history, experiences with cancer and genetic testing, motivations to pursue genetic testing and subsequently disclose genetic test results, information sharing patterns, healthcare provider response, and participants' emotional support systems. The interviews were transcribed in their entirety, coded, and analyzed based on grounded theory. Results: Eighteen transcripts were used for the analysis. Results can be classified into 5 main themes. Participants (n=8) were most concerned about cancer risk for their children and female family members, and most (n=11) mentioned HBOC provides them increased personal awareness, but has a negligible impact on their life overall (n=9). Men (n=11) were interested in a male focused support group to discuss HBOC and gain knowledge and information. Participants (n=9) took on active and open communication roles with family members and health care p (open full item for complete abstract)

Committee: Leigha Senter MS, LGC (Advisor); Robert Pilarski MS, LGC (Committee Member); Doreen Agnese MD (Committee Member) Subjects: Gender; Genetics

Master of Science, The Ohio State University, 2016, Genetic Counseling

Women with a BRCA1 or BRCA2 mutation have a 40-74% breast cancer risk and 11-46% ovarian cancer risk by age 70. Due to this elevated risk, it is recommended that mutation carriers have increased breast screening and a risk-reducing salpingo-oophorectomy. They are also given the option to have a risk-reducing mastectomy. The process of deciding if and when to undergo prophylactic surgery has been found to be a complex and difficult process for many carriers. To evaluate the decision-making process, recorded interviews were conducted with 20 BRCA1 and BRCA2 mutation carriers. Information about the cancer risk management decision-making process was analyzed using grounded theory. Factors involved in decision-making, ease of decision-making, and individuals involved in the decision-making process emerged as major themes. Mutation carriers who have not had breast cancer (previvors) were found to have a more difficult time coming to a cancer risk management decision than women with a breast cancer history. Physicians were often discussed as being an integral part of the decision-making process by providing support and management recommendations. Family members and other mutation carriers filled a similar role during the decision-making process by providing decisional and emotional support for carriers. Genetic counselors were short-term providers of risk information and management recommendations for this study population. If a carrier was not receiving the information or support she needed from one of these groups, she often turned to another party, most commonly a healthcare provider. Thus, data from this study suggests that previvors or mutation carriers struggling with the risk management decision-making process may need additional support and information to assist them during this process. It is important that healthcare providers are educated about risk-management strategies for HBOC and work together to best help mutation carriers through the decision-making proces (open full item for complete abstract)

Committee: Amanda Toland PhD (Advisor); Shelly Hovick PhD (Committee Member); Leigha Senter-Jamieson MS, LGC (Committee Member) Subjects: Genetics; Health Care; Health Care Management

Master of Science, The Ohio State University, 2016, Genetic Counseling

Women with hereditary breast and ovarian cancer syndrome (HBOC), caused by mutations in BRCA1 or BRCA2, have increased lifetime risks of certain cancers, including breast and ovarian cancers. Lifetime cancer risks are presented to BRCA mutation carriers during genetic counseling, often with the addition of statistical figures and graphs. This study examines how factors such as demographic characteristics, health numeracy, graph literacy, and HBOC knowledge affect BRCA mutation carriers' preferences for and understanding of different cancer risk estimate formats, including line graph, bar graph, icon array, and text-only. An anonymous online survey was completed by 82 BRCA mutation carriers that assessed attitudes, comprehension of, and preferences for the cancer risk estimate formats. Participants best understood lifetime cancer risks when presented using the text-only format, but preferred their lifetime cancer risk be presented graphically. The line graph was the most preferred and most easily understood graphical format for presenting lifetime cancer risks. Increased comprehension of the line graph was associated with higher graph literacy (p<.05), while increased comprehension of the bar graph and icon array were associated with higher health numeracy (p<.05). Results suggest that when presented with lifetime cancer risks in genetic counseling, BRCA mutation carriers may benefit most from text and graphic displays, particularly a line or bar graph, to help describe their risks. Line graphs may be more effective for patients with higher graph literacy, whereas bar graph may be more effective for patients with higher health numeracy.

Committee: Shelly Hovick PhD (Advisor); Leigha Senter-Jamieson MS (Committee Member); Kevin Sweet MS (Committee Member) Subjects: Genetics

MS, University of Cincinnati, 2004, Allied Health Sciences : Genetic Counseling

This study characterized women's responses to the direct-to-consumer advertising campaign for BRACAnalysisreg; hereditary breast and ovarian cancer susceptibility testing. The study assessed women's intent to pursue testing before and after viewing the commercial and identified where women would seek out information. Pre- and post-test questionnaires assessed family history of breast cancer, breast cancer anxiety and risk perception, as well as the likelihood that women would pursue genetic testing for breast cancer risk. After viewing the advertisement, 73% of women reported interest in information about BRACAnalysis®. Overall, 76% of women reported that they would seek information on BRACAnalysisreg; testing from their OB/GYN and 37% would go to their family doctor. Being that OB/GYNs may soon be faced with an increase in questions about and requests for BRACAnalysisreg; testing, it is imperative that an educational plan be set in place to train OB/GYNs further about the genetics of breast and ovarian cancer.

Committee: Jennifer Gamm (Advisor) Subjects:

Doctor of Philosophy, The Ohio State University, 2011, Biochemistry Program, Ohio State

The main goal of the research discussed in this dissertation is to create effective and safe cellular hemoglobin (Hb)-based oxygen (O2) carriers (HBOCs) for use in transfusion medicine. This dissertation focuses on the development and scale up production of cellular HBOCs composed of novel lipid formulations and polymers. The research concentrates on the development of cellular HBOCs over acellular HBOCs and abiotic O2 carriers, owing to their various advantages, such as, longer shelf-life, high Hb content, increased circulation half-life, biocompatibility, and absence of vasoactivity and hypertension in vivo. This dissertation presents various novel large scale methods for the production of cellular HBOCs. The methods discussed are very efficient as HBOCs are produced with very reproducible biophysical properties. All the methods are designed with cost effectiveness in mind. The methods presented are simple and easily scalable to an industrial scale. In addition, they eliminate the use of energy consuming equipment/methods used in the production of other cellular HBOCs. Moreover, all the scale up methods are designed to satisfy important design criteria required to develop an effective O2 carrier, such as, high Hb content, suitable biophysical properties to ensure proper O2 delivery to tissues and organs and absence of any free Hb in the dispersions. This dissertation also outlines detailed insights on the role of the intracellular diffusion barrier, as a result of Hb encapsulation, on binding/release of gaseous molecules to Hb. It emphasizes the importance of the cellular membrane and Hb encapsulation, as well as the pivotal role it plays in regulating O2 delivery or consumption of nitric oxide (NO) by Hb in cellular HBOCs. In addition, the ex vivo experiments in rat arterial segments discussed in this dissertation provide for the correlation between O2 delivery and NO consumption to Hb-induced vasoconstriction by using exogenous and endogenous sources of NO. (open full item for complete abstract)

Committee: Andre Palmer PhD (Advisor); Paulaitis Michael PhD (Committee Member); Kuppusamy Periannan PhD (Committee Member); Magliery Thomas PhD (Committee Member) Subjects: Alternative Medicine; Biochemistry; Chemical Engineering

Doctor of Philosophy, The Ohio State University, 2011, Chemical and Biomolecular Engineering

The frequent shortages, risks of disease transmission, and storage issues associated with donated blood illustrate a significant demand for a red blood cell (RBC) substitute. Such a substitute should be able to effectively transport oxygen throughout the body with minimal side effects. Several hemoglobin-based oxygen carriers (HBOCs) have been developed and clinically tested, but they have all caused severe side effects. The problems associated with these HBOCs may all be attributed to removing hemoglobin from the RBC. Therefore, this work focuses on the use of the extracellular hemoglobin of the earthworm Lumbricus terrestris (LtEc) as a new class of HBOC. Since earthworms lack RBCs, their hemoglobin is freely dissolved in the bloodstream and has already adapted to solve many of the challenges facing modern synthetic HBOCs. It has a lower rate of oxidation, avoids harmful side reactions with nitric oxide (NO), and it is extremely stable. We have developed a novel purification technique to highly purify large amounts of LtEc at costs that are comparable to donated blood. The LtEc product also transports oxygen similarly to human blood. Transfusion of LtEc into hamsters does not elicit the harmful side effects observed with other HBOCs and preliminary studies have not revealed any immune or allergic reactions in vivo. Therefore, this work shows that LtEc might be an effective and safe oxygen carrier that warrants further study and suggests the need for a paradigm shift in the HBOC field from cellular to extracellular hemoglobins.

Committee: Andre F. Palmer Ph.D. (Advisor); David Wood Ph.D. (Committee Member); Jessica Winter Ph.D. (Committee Member); Boris Mityagin Ph.D. (Committee Member) Subjects: Biology; Chemical Engineering