Interdisciplinary – Page 9 – UROP Spring Symposium 2021

Interdisciplinary

Analyzing CRISPR-Cas9 through Genomic Deletions

Background: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is a technology used to edit genomes at very high precision. It enables precise editing of genomic loci with a RNA-guided CRISPR associated protein 9 (Cas9) nuclease that can cleave the targeted DNA complementary to a guide RNA (gRNA). Precise gene editing via CRISPR-Cas9 has great potential in treating inherited diseases (e.g., cystic fibrosis) or correcting genetic defects.

Objective: To test the specificity and function of the CRISPR-Cas9 in a proof-of-principle experiment, we used two human Emx1 (hEmx1) gene-specific gRNA sequences (3.1+4.1) to guide the Cas9 enzyme for the deletion of the targeted region in human 293AD cells.

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Relationship Between Depression and Subjective Executive Functioning in Parkinson’s Patients

In addition to physical symptoms, Parkinson’s patients are often diagnosed with mental complications including depression and cognitive dysfunction. However, little is known about the correlation between Parkinson’s patients’ depression and subjective cognitive skills, including executive functioning. This is important because executive dysfunction can cause difficulty in Parkinson’s patients’ ability to complete daily tasks. The main goal of this research is to examine how mental factors such as depression impact a Parkinson’s patient’s subjective executive functioning. In order to compare these two variables, archival clinical data was analyzed in a sample of 175 patients with Parkinson’s disease (PD; N(men) = 120, M(age) = 64.41, SD = 8.58). The Geriatric Depression Scale – Short Form (GDS-SF) was used to measure depression, the Frontal Systems Behavior Scale (FrSBe) was used to measure subjective executive functioning, and the Unified Parkinson Disease Rating Scale (UPDRS) Part III was used to measure PD severity. Pearson’s correlation revealed a significant relationship between the GDS and FrSBe scores, such that higher depression scores correlated with higher self-reported executive dysfunction, R = .42, p < .001. The partial correlation revealed that this relationship remained significant while controlling for UPDRS Part III, R = .45, p < .001. Findings were consistent with this study's hypothesis, such that Parkinson's patients with greater depression were more likely to self-report executive functioning problems. A possible explanation for this is that depressed Parkinson's patients are more likely to have negative self-perceptions, which may cause them to rate their executive functioning skills more poorly.

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Examining how COVID has affected African American women with Hypertension

Forty seven percent of African American women are living with hypertension, which increases their risk of contracting COVID-19. We asked 100 African American women living with hypertension (ages 21 to 64) to complete a questionnaire using qualtrics. Using a cross sectional descriptive survey, participants were asked to self-report their blood pressure during COVID. A portion of the survey was used to raise the questions of how paticipants have been maintaining their health through eating and exercise patterns as a result of the virus. Sixty one percent of participants reported that they have not had to change the way that they manage their blood pressure since COVID-19. Participants that reported that they did have to change the way that they managed their blood pressure, ensured that they have had to exercise more, practice healthier eating habits, and that they haven’t seen their physician since COVID-19. In addition, the majority of participants did not lose their job or income due to the “Stay at Home” mandate, nor are they concerned about their ability to get their medications and supplements used to control their blood pressure. Our findings raise the question regarding if participants are considering all factors that have been impacted in their lives by COVID and how participants have been managing their hypertension in the first place.

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The Facial Grimace Test in the Detection of Hyperacusis in Mice: Sex Differences

Hyperacusis is an auditory disorder characterized by an increased sensitivity to sounds, which can result reduced auditory pain threshold. Sex differences in the induction of hyperacusis are not well understood. Reports of hyperacusis in humans are higher in men (Paulin et. al., 2016), under the assumption that males have a higher exposure to high-intensity noise resulting in hyperacusis. However, a noise-exposure-induced threshold shift is not necessary for hyperacusis. Due to variability in noise-exposure across life-span and reporting, it is difficult to accurately assess sex differences in the prevalence of hyperacusis in humans. In this study, the same level of noise-exposure was administered to male and female mice, followed by behavioral assays to determine the hyperacusis status of each animal. Preliminary data in the conditioned place-aversion test suggest that there are sex differences in the induction of hyperacusis in mice. To support these results, the facial grimace test was adapted to assess the auditory pain threshold in mice. The facial grimace can be characterized by previously established grimace scales. Data from the facial grimace test in both male and female mice was analyzed in order to determine whether a sex difference was present between mice with and without hyperacusis. This assay confirmed the presence of hyperacusis in male but not female mice. Future studies are needed to investigate the underlying genetic predisposition to the development of noise-induced hyperacusis. Paulin, J., Andersson, L., & Nordin, S. (2016). Characteristics of hyperacusis in the general population. Noise & health, 18(83), 178-184. https://doi.org/10.4103/1463-1741.189244

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Tissue crosstalk in joint injury

Post-traumatic osteoarthritis (PTOA) is a debilitating disease that develops as a result of a joint injury. Overactivation of the Wnt signaling pathway has been implicated in progression of PTOA, however very little is known about this. This work is among the first to study Wnt signaling mechanisms in PTOA. R-spondin 2 is a secreted matricellular protein that functions as an agonist for the Wnt pathway, promoting Wnt signaling. Our preliminary data show that R-spondin 2 is greatly induced in the synovium in a mouse model of PTOA. We also showed that R-spondin 2 is secreted into the synovial fluid after joint injury. This deregulation of R-spondin 2 promotes over-stimulation of the Wnt pathway, which we hypothesize is an important mechanism in joint deterioration. Our aims are to understand the cellular and molecular mechanisms underlying R-spondin 2 and Wnt signaling in PTOA, and how we can manipulate these to inhibit further joint degeneration. To study gene expression in joint-resident cell types, we first analyzed bone marrow stromal cells – primary cells cultured from mice and differentiated into osteoblasts, a cell type present in the joint and relevant to PTOA progression. We isolated RNA, converted it to cDNA, and performed qPCR to analyze gene expression. In parallel, we employed microscopy to study joint pathology over the course of PTOA. Further investigation is now warranted, from which we hope to better understand the cellular and molecular mechanisms of Wnt signaling in PTOA, with the ultimate goal of developing novel therapeutics for treating PTOA.

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Evaluating Novel Cancer Drugs and Drug Delivery in Preclinical Studies

Cancer prevention and treatment in the United States has seen many advances over the last few years, however, breast cancer continues to be one of the most widespread diseases, affecting the lives of approximately one in every eight American women. There is a pressing need for a more effective and less invasive treatment for breast cancer, specifically those cancers that originate in the epithelial cells of the milk ducts or lobules, also known as adenocarcinomas. Heat shock protein 90 (HSP90) has become an important target of many drug treatments. At the molecular level, over-expressed signaling proteins that lead to uncontrollable cell growth are stabilized by HSP90. Recent in vitro studies of the specific HSP90 inhibitors, KU757 and KU758, suggest this drug is effective in treating certain types of cancer, such as thyroid cancer and triple negative breast cancer respectively. This study aims to investigate the efficacy of KU757 and KU758 in treating the breast adenocarcinoma cell line called MCF7. Cell proliferation will be measured by MTS assay, and drug efficacy will be analyzed by calculating inhibitory concentration 50 values.

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System for Opioid Overdose Surveillance (SOS): Genesee County Project

Background: The opioid epidemic has grown across Michigan with very few areas having access to real-time data on opioid overdoses. A project was conducted by the CDC-funded University of Michigan Injury Prevention Center and Michigan High Intensity Drug Trafficking Areas (HIDTA) to develop a system for near real-time opioid surveillance. This system, called the System for Opioid Overdose Surveillance (SOS) was designed to inform data-driven opioid overdose prevention and response efforts with the goal of reducing overdose injuries and fatalities. Methods: In a Genesee County project, community stakeholders composed of public health, public safety, and community outreach service providers were gathered for a focus group, and then interviewed to evaluate how near real-time opioid overdose data can inform prevention work and barriers to prevention efforts. Stakeholders received SOS data reports for their jurisdiction on a regular basis for eight weeks, and their feedback was solicited over time through a survey. A final focus group emphasized the mobilization of data-driven coordinated community responses through near real-time reports. Data was analyzed to create a coordinated community response toolkit. Results: Findings suggest SOS is being used to impact local planning and responses to opioid overdoses. One major finding from the Genesee County project was the application of zip codes in aiding community stakeholders. The use of zip codes, one of many data visualizations in the SOS, allowed for stakeholders to pinpoint locations most affected by opioid overdoses within the community. Conclusions: Through the Genesee County Project, the SOS dashboard helped aid in creating “best fit” strategies for specific areas within larger communities. The Genesee County Project also showcases how near real-time data surveillance can benefit communities. Keywords: Opioids, Surveillance, Genesee County

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Worlding Medieval and Early Modern Iberia

The Iberian Peninsula, home to Spain and Portugal, is rich in the history of the Spanish, Portuguese, Arabic, and Hebrew languages; Muslims, Jews, and Christians’ tumultuous histories; and many more aspects of intellectual literature and culture. Through this research project, which resulted in the creation of a mapping website to study the medieval text, “El libro de conosçimiento de todos los rregnos,” students will be able to learn about the world through the lens of a medieval traveler. The study utilizes “El libro de conosçimiento de todos los rregnos” in order to gather information about the wide variety of locations visited by the anonymous author in the fourteenth century. Qualitative data regarding names of regions and towns; persons of interest; and the coordinates of the aforementioned places are recorded. In addition, mythological creatures and locales are also recorded when named by the traveler. Analyzing this project leads a reader to discover the environment of the early 1300s, mapping the travels from Ireland to Italy to Istanbul. Shown through the map-based website the researchers created, students and pedagogists alike may better examine the history of the Iberian Peninsula. As “El libro de conoçimiento de rregnos” has not been studied in great detail compared to other texts from the same time period and locations, the website provides a fresh look for students of history, Iberian culture, and language.

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Worlding Medieval and Early Modern Iberia

Project: Worlding Medieval and Early Modern Iberia Student Signature: Hannah Cohen Mentor Signature: Luis Miguel dos Santos Motivation/problem statement: As it may be very difficult for students to read and analyze Spanish literature from Medieval times, the research conducted in this project will make the process simpler and easier to understand. Turning something that makes little sense in modern day language to something that does make sense will eliminate the confusion students have. Methods/procedure/approach: Analyzing Spanish literature texts from older times is how the results get drawn up. “Libro de Conocimiento” is the main text that contains information on people, places, and more from the times of the past. There are also other shorter stories relating to similar topics that provide additional findings. Results/findings/product: After picking out the important information about the past and completing the research on what will help current Spanish students understand what occurred in these past locations, a website is to be created. This website will display a map with the most important places, people, and might add some pictures and audios along the way to allow Spanish students to get full knowledge of what information they are to get from Medieval Spanish literature. Conclusion/implications: There is to be a greater understanding of Spanish literature, related to old places and people, for students. The website will be in great value as a guide in comprehension for those who are taking a Spanish literature course about the past.

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A 5-year Longitudinal Observational Study of Management of Patients with Hepatocellular Carcinoma

Hepatocellular Carcinoma(HCC), a liver cancer that is linked with hepatitis A and B which are caused by liver cirrhosis, is an increasingly prevalent cancer that affects many people across the globe, majorly in the United States, Asia, and Africa. Treatments and remedies are currently very limited with nothing holding the capabilities of fully healing affected patients. The purpose of this research is to record and analyze the treatments currently used over the course of five years and create a database for future patients who face the cancer. Patients are searched for by project managers and are selected if they have HCC and commit to the project. The chosen patients undergo a health background check and provide documentation of their treatment and checkups every three months for a year and every six months for four more years. Over the course of five years, the patients are monitored and their medical documentation is used as data for the database. The database will allow physicians to categorize the effects of different treatments with the different types of patients to find the best solutions for future patients. After five years, the goal is not necessarily to find a cure or remedy, but rather to have an established database of information to allow doctors to better treat patients with HCC and create research further creating potential for a cure or remedy. The database’s main goal is to allow other researchers to take advantage of the material published and explore different methods of HCC treatment. Currently, the project remains partially complete and will require several years before the database becomes available for publication.

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