Nia Acharya, Krishna Gundabolu, Vijaya R. Bhatt, Alex B. Nester, Laxmi Narayana Buddharaju, Pierre Fayad, Daniel Surdell, and William E. Thorell
CVST is a rare location of thrombosis involving Dural/ cerebral venous sinuses. It affects around 5-10 people per million population annually.
It is an uncommon but life-threatening form of stroke affecting younger individuals. Therefore, identifying and treating in a timely manner is critical.
Rarer thrombotic disorders like paroxysmal nocturnal hemoglobinuria (PNH) or Janus Kinase 2 (JAK2) mutation positive myeloproliferative neoplasms (MPN) can rarely present with CVST. It can also present during pregnancy for the first time.
Diagnosis is often established by Computed Tomography (CT) or Magnetic Resonance Imaging (MRI).
Infections, certain medication use (asparaginase or birth control pills) could lead to CVST. Patients often present with headaches, seizures or neurological deficits.
Management is often with systemic anticoagulation despite intraparenchymal hemorrhage. Reducing intracranial pressure by invasive approaches is sometimes needed.
Anastasia Bernal, Borum Ryu, and Grinu Mathew
Prostate cancer (PC) is the second leading cause of death due to cancer among men in the United States. Metastatic disease is commonly seen in sites such as lymph nodes, bone and in visceral organs like lung and liver. The 5 year survival rate of patients with primary PC is almost 100%, however, this survival rate is drastically reduced to 30% in patients with metastatic disease. Therefore there is an unmet need to identify genes associated with metastatic progression and develop therapeutic strategies to increase survival rate in patients with aggressive disease. The loss of TAM kinase family receptor AXL is observed in metastatic sites, suggesting that AXL is a potential tumor suppressor in metastatic PC. Data from our laboratory further supports that loss of AXL gene is significantly co-deleted with tumor suppressors PTEN and TP53 in patients with metastatic PC. To test the functional role of AXL gene in metastasis, we generated lentiviral plasmid constructs to express a) doxycycline based inducible knockout of AXL (CRISPR-Cas9 gene editing, and b) overexpression of full length and domain mutants of AXL. Transformation efficiency indicated effective cloning, and long read sequencing with Oxford Nanopore technology confirmed that the cloned inserts were successful. A plasmid map was created for each construct using Snapgene software, and bioinformatic analysis of AXL gene was performed using NIH BLAST. Next, we transduced PC cell lines with lentivirus expressing the designed vectors. The results indicate that overexpression of full length Axl (mouse) in LNCaP-FGC cell line exhibited a rounded morphology with Axl expression (EGFP tag) confined to the plasma membrane and cytoplasm. Interestingly, cells expressing the kinase dead mutant of Axl protein (K561R in the ATP binding site) showed an elongated morphology with distinct plasma membrane localization. Future directions for research include a migration assay to probe the metastatic potential of cells with KO or overexpression of Axl. We will also compare full length vs kinase dead/domain mutants to see the rate of migration. Furthermore, we will also test the efficiency of doxycycline inducible expression of single guide RNAs to knockout AXL in PC cell lines. These systems can now be utilized to test our hypothesis that AXL loss induces metastasis in PC.
Analysis of COVID-19 Pandemic Disease Metrics in Nebraska Counties with Large Meat Processing Plants
Nyah D. Chambers; James Lawler MD,MPH; and Sara Donovan MPH
This poster analyzes COVID-19 disease metrics of counties with large meat processing facilities to counties with small or no meat processing facilities based on different time frames of the COVID-19 pandemic.
Optimization of Tick Attachment and Detection of B. Burgdorferi Transmission into In Vitro Generated Skin Rafts
Carley M. Conover and Amanda J. Brinkworth
Borrelia burgdorferi (B. burgdorferi) is a Lyme Disease causing pathogen that is transmitted via tick-bite by blacklegged (deer) ticks (Ixodes scapularis). These ticks tend choose hosts such as rabbits and mice to feed on, sometimes transmitting harmful pathogens to these animals such as B. burgdorferi, which cycles between rodents and large animals to complete its enzootic cycle. Humans are accidental dead-end hosts for B. burgdorferi. Those infected with B. burgdorferi can develop Lyme Disease, which includes symptoms such as neurological, cardiac, and joint-related complications which are exacerbated by increased feeding time on the host1. Due to the many negative affects Lyme Disease can have on humans, there is a need for medicinal options that could prevent the accumulation of the bacteria at the tick bite site or inhibit the ability of ticks to feed on the human host. We rationalize that a human skin model to study tick-human skin interactions and possible therapeutic screening is needed as there are currently no cost-effective or high-throughput options.
Our in vitro generated skin rafts will enable us to study tick feeding habits and B. burgdorferi transmission comparable to use of in vivo animal models, but more applicable to human infection. We hypothesize that, as B. burgdorferi-positive ticks feed on the rafts, they will transmit the bacteria into the epidermal layer which can later be extracted and quantified. Once these skin rafts have been optimized for maximal tick feeding and B. burgdorferi transmission, they can then be used in high-throughput experiments to find therapies that could modify the bacteria or tick interactions with the host and prevent development of Lyme Disease.
Coding Severe Behaviors in Children with Autism Spectrum Disorder to Train Machine Learning Algorithms
Toluwa Davies, Walker Arce, Jordan DeBrine, Katelyn Malick, Carter Welch, James E. Gehringer, Seth Walker, and Amanda Zangrillo
Thermodynamic Contributions of Deoxyuridine Loops to the Folding of DNA Straight Hairpin Loops with d(GCGC/GCGC) Stems
Benjamin G. Dodson and Luis Marky
The global stability of DNA molecules depends on base stacking, base-pairing, ionic and hydration contributions. To understand the mechanisms that govern the many biological roles of nucleic acids, it is essential to have a complete physical description of the folding of nucleic acids, including ion and water binding. We used a combination of temperature-dependent UV spectroscopy and differential scanning calorimetry to investigate the effect of the loop length on the unfolding thermodynamics of a set of DNA stem-loop motifs with the following sequence: d(5’-GCGCUnGCGC), where n = 3, 5, 7, and 9.
The folding of each hairpin loop resulted in a monophasic transition that was monomolecular. The favorable folding of each hairpin (negative ∆Go) in this set resulted in a thermodynamic compensation of a favorable enthalpy with an unfavorable entropy contributions, enthalpy contributions correspond to formation of base-pair stackings while the unfavorable entropy contributions correspond mainly to the immobilization of counterions, as the length of the uracil hairpin loop increases, the stability of the hairpin decreases (the free energy term becomes more positive), due to a less favorable entropic contribution because more counterions are binding. Relative to a similar set of thymine hairpin loops, we measured less favorable free energy terms because of the lower thermal stability of the uracil hairpins (lower TM’s) and their lesser impact of ion binding. Supported by Grant MCB-1912587 from the NSF.
Alexander C. Eischeid, Sylvain Bernhardt PhD, Arvind Natarajan MS, and Hani Haider PhD
Computer-aided surgery capabilities for arthroplasty interventions emerged in the late 1990s with limited capabilities and use in the field. The goal of computer-aided surgery in the field of arthroplasty remains to reduce the need for excessive drilling tissue damage to the surgical site by reducing the need for cutting guides and related jigs. In order to assess the capabilities of computer-aided surgery systems in terms of both their accuracy and precision, the development and adherence to an industry standard for testing is necessary. A phantom device with divots of a known coordinate location was used in conjunction with proprietary software to assess the accuracy and precision of the complete surgical system. Measured coordinate data of a single point using the proprietary software was transformed according to the balloted ASTM standard for the generation of an accuracy and precision report. Results indicate 0.255 mm accuracy and sub-millimeter precision under conditions most similar to an operating room. Functional extreme tests indicate a loss of performance, leading to maximum decreased accuracy of 1.71 mm during standard orientation and 4.32 mm during extreme orientation of the phantom. The results suggest the tracking and software system meet manufacturer data under standard orientation and location conditions yet experience an expected significant loss of ability in extreme conditions. These loses in capability may lead to inaccurate alignment of tools and implants when using the proprietary computer-aided surgery software. Additional research is needed to determine the effect of larger reference frames with additional tracking points. Additionally, development of software to limit data return when near functional extremes will improve ease of use of the system.
Hossam Elaraky, Aarfa Queen, MD Shafikur Rahman, Angie Carmona, Hamdan Alrefaei, and Paul C. Trippier
Neurolysin (Nln) is a neutral metalloendopeptidase, which is an enzyme that breaks down peptide bonds in amino acids whilst ultilizing a metal catalyst1. Nln inactivates several neuro/cerberotoxic neuropeptides such as neurotensin, substance P, and bradykinin2. These neuropeptides can cause harm to the brain as they respond to inflammation during an ischemic stroke3,4. The peptides generated by Nln are neuro/cerebroprotective which aids in its ability to protect the brain in an ischemic stroke victim2. The goal of our lab is to discover a potent neurolysin activator in order to develop a drug that protect victims from brain damage and disability following an ischemic stroke. To begin this process, our lab successfully designed, synthesized, and established the structure-activity relationships of several potential neurolysin activators via a preliminary neurolysin activation assay on several targeted compounds. Secondary amine analogs were created and synthesized to test their neurolysin activation potential. The goals of this experiment are to resynthesize these compounds to gather additional NMR and HPLC data and to optimize the reaction to obtain a pure compound with a substantial yield.
Gabrielle H. Estep, Ashley Peer, Mohammad Salimi, Michael Wiley, and Catherine Pratt
Detection of Adenovirus, Aichi virus, Influenza, Cytomegalovirus, respiratory syncytial virus, and Hepatitis A in Nebraska wastewater using RT-PCR. Extraction of virus nucleic acid was done using Dynabead enrichment and MagMAX wastewater extraction kit.
Intracellular Parasite Toxoplasma Exploits the Unfolded Protein Response to Acquire Mitochondrial Metabolites
Ethan J. Funke and Leonardo da Silva Augusto
Toxoplasma gondii is an obligate intracellular parasite that reconfigures its host cell to promote pathogenesis. Toxoplasma is able to infect virtually all warm-blooded animals, and it is estimated that this parasite has infected nearly 2 billion people globally and approximately 25% of the US population. Curiously, upon infection Toxoplasma recruits the host cell’s endoplasmic reticulum (ER) and mitochondria into close proximity to the parasitophorous vacuole (PV), although the reasons for these high affinity interactions are not completely understood. It has been shown that Toxoplasma induces host mitochondria elongation to acquire fatty acids and establish a niche for itself. Our exciting new data showed that through the host ER-PV association, Toxoplasma actively induces the unfolded protein response (UPR), leading to PERK activation. Furthermore, new data suggests that PERK activation coordinates mitochondrial elongation. Therefore, we hypothesize that PERK activation induces mitochondrial elongation in Toxoplasma infected cells. Using immunofluorescence, we determined whether PERK activation plays a role in mitochondrial elongation induced by Toxoplasma infection. First, we confirmed that the Toxoplasma PV interacts with host mitochondria and ER. Next, we found that Toxoplasma infection promotes mitochondrial elongation and by using a pharmacological approach, we were able to inhibit PERK, which significantly decreased mitochondrial elongation. Our study has identified a novel mechanism used by Toxoplasma to induce mitochondrial elongation in order to acquire fatty acids, providing new insights into strategies for treatment of toxoplasmosis.
Phosphatidylethanolamine Methyltransferase Deficiency Exacerbates Acute Alcohol-Induced Liver Injury
Jessica A. Gardner, Madan Kumar Arumugam, Srinivas Chava, Sathish Kumar Perumal, and Kusum K. Kharbanda
Alcohol-associated liver disease (ALD) is a global burden of healthcare and remains a major cause of morbidity and mortality worldwide. ALD includes a spectrum of injuries that progresses from hepatic steatosis, alcoholic hepatitis to alcohol-associated cirrhosis and even hepatocellular carcinoma with continued alcohol misuse. The development of ALD depends on several factors, including genetics. The liver enzyme phosphatidylethanolamine methyltransferase (PEMT) catalyzes three sequential methyl transfers to phosphatidylethanolamine, generating phosphatidylcholine (PC). The PC generated with PEMT-mediated catalysis is preferentially used in very-low-density-lipoprotein (VLDL) assembly and is required for its normal biogenesis and secretion (1-3). Alcohol affects the methylation potential and impairs PEMT activity, which by inhibiting VLDL synthesis contributes to the development of hepatic steatosis. Polymorphisms in the human PEMT gene causing loss of function confer susceptibility to metabolic-associated steatohepatitis.
Based on these considerations, we hypothesized that PEMT deletion would exacerbate alcohol-induced liver injury.
Animal Handling and Diet:
Male and female wildtype (WT) and PEMT knock out (KO) mice (12 weeks of age) were subjected to ethanol binge feeding model. The animals were gavaged with maltose dextrin or ethanol (5g/Kg BW) twice, 12 hours apart. The mice were euthanized eight hours after the second dose, where the blood and liver were collected for the following analyses:
AST and ALT levels: Serum AST and ALT were analyzed using a VITROS 5.1 FS Chemistry System.
Hepatic histopathology: Neutral-buffered formalin fixed liver sections stained with hematoxylin & eosin (H & E) and picrosirius red were imaged using a Keyence BZ-810 microscope.
HPLC Analysis: Liver tissues were homogenized in 0.5N perchloric acid and subjected to HPLC analysis to determine S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) levels (3,4) The SAM:SAH ratio, or methylation index, was calculated, as detailed (3,4).
Triglyceride Quantification: Lipids were extracted by Folch method (6) and triglyceride levels were quantified using the Thermo DNA Kit (3,4).
Enzymatic Activity: Lysosomal acid lipase and proteasome activities were determined in liver homogenates, as detailed (7)
Statistical Analyses: Data are expressed as mean values ± standard error (SE). Values not sharing a common subscript letter are statistically different, p < 0.05.
We found deletion of PEMT exacerbates acute alcohol-induced liver injury in both males and females as evidenced by:
•Increased AST and ALT levels
•Increased fat accumulation by histopathological assessment
•Decreased SAM levels causing a reduction in the methylation potential •Increased hepatic triglycerides •Decreased lysosomal acid lipase activity •Decreased proteasome activity
Concordance of Cross-Sectional Imaging and Adrenal Venous Sampling Results for Patients with Surgically Treated Primary Hyperaldosteronism
MaKayla M. Gordon, Whitney Goldner, Anupam Kotwal MBBS, Anery Patel, Jessica Shank, and Abbey Fingeret
Adrenal venous sampling (AVS) is used to distinguish unilateral from bilateral aldosterone hypersecretion as a cause of primary hyperaldosteronism (PHA). This distinction is critical because unilateral disease is treated, and often cured, by adrenalectomy, whereas bilateral hypersecretion should be managed medically.
We performed a retrospective cohort review of adult patients undergoing index adrenalectomy for PHA at the University of Nebraska Medical Center from July of 2013 to June of 2022. Clinical and pathologic variables were assessed including patient age at surgery, sex, race or ethnicity, body mass index, systolic and diastolic blood pressure, number and type of antihypertensive medications pre- and post-operatively, potassium level and supplementation, PAC, PRA, ARR, imaging findings, adrenal venous sampling results and concordance of imaging findings with AVS and surgical outcomes. Statistical analysis was performed with Mann Whitney U and chi-squared Fisher’s exact using STATA version 17.
In our cohort, 21 patients were identified who underwent adrenalectomy for primary hyperaldosteronism. Of these, 2 patients did not have any imaging findings and 19 were image localized. For patients with image localization AVS was concordant in 9, discordant in 4, and nondiagnostic in 6. For both patients without image localization the AVS was lateralizing. The overall discordance between imaging results and AVS was 40%. The only significant difference between patients with concordant and discordant results was the aldosterone level with concordant patients having a mean of 58ng/dL compared with discordant patients 23ng/dL (p = 0.0022). There was a significant reduction in antihypertensive medications in the entire cohort from a mean of 3.2 medications to 1.2 medications (p < 0.001).
Conclusions and Future Directions
In this cohort, 40% of patients with selective AVS had discordant imaging and AVS results. Preoperative plasma aldosterone concentration was positively associated with concordance, with higher PAC more likely to have imaging and AVS concordance. Overall, hypertension was significantly improved following adrenalectomy for PHA with a median decrease of 2 antihypertensives. Our results support the recommendation to perform AVS on all candidates for adrenalectomy for PHA. Further study is warranted to identify factors associated with discordance.
Kaitlyn M. Hart and Amanda Marshall
The data collected in this project will be used as a part of a multi-center study conducted by Boston Children’s Hospital to assess if the software Etiometry® can be used to predict extubation success in patients in the CICU. After compiling the data from EPIC, the goals of our analysis were to find any associations with extubation failure rates. Trends in this data could help identify potential areas of focus to mitigate extubation failures. Some specific hypotheses that were explored included the associations between extubation failure and delayed sternal closure, increased length of stay, length of post-op stay, ECMO, and death. The similarity between CHMC’s rate of extubation and what is being reported nationally was also considered. Data was collected by reviewing patient charts in EPIC and analyzed using Chi squared and the Student’s T-test where appropriate. This study found that neonatal status and escalated respiratory support in within 48 hours trended towards an association with extubation failure. It was also found that extubation failure trended towards an association with death. Within the collected data, there was not one overarching diagnosis that was more common in failed extubation encounters than successful encounters. While the conclusions from the data collected in this study are limited by the sample size, this data will benefit the multi-center study conducted by Boston Children’s Hospital in assessing the accuracy of Etiometry® in predicting extubation success.
Natalie Holding, Mohsin Raza, Vimla Band, Bhopal C. Mohapatra, and Hamid Band
ECD, Ecdysoneless protein, is evolutionarily conserved. It was identified as a human homologue of Drosophila ecdysoneless, interacting with HPV16 E6 in a yeast two-hybrid assay. ECD has been proven to be essential for cell cycle progression from G1 to S phase, mitigating endoplasmic reticulum (ER) stress, and embryogenesis. ECD KO mice are embryonic lethal as it halts the cell cycle at G1. ECD interacts with p53 and Rb. ECD also associates with pre-mRNA splicing factor PRPF8. ECD is over-expressed in breast, pancreatic, gastric, and Human Papilloma-driven cancers and is correlated with shorter patient survival. ECD interacts with co-chaperone complex R2TP involved in protein assembly and folding. ECD over-expression has been proven to increase oncogenesis regulated by c-MYC in a recent mice model. Biochemical analyses showed ECD to have a role in mRNA splicing and nucleus to cytoplasm export. The purpose was to determine if there is an interaction between ECD and EIF4A1 and determine ECD’s effect on global translation using HEK293T (human embryonic kidney) and SUM159 (breast cancer) cell lines. Overall, the interaction between ECD and EIF4A1 supports a novel mechanism by which ECD protein regulates eukaryotic mRNA translation. This mechanism may contribute to the resistance of cancer cells over-expressing ECD to the translation-inhibitory effect of endoplasmic reticulum stress prevalent in tumors.
Representation and Outcome of Catheter Ablation for Treatment of Atrial Fibrillation Among Patients with Obesity: A Systematic Review of Randomized Control Studies
Eh M. Khaing, Danielle Dircks, Ahmad Aroudaky, Muaaz Almerstandi, James Aguto, Jmaylia Mimms, William Schleifer, Jason Payne, Arthur Easley, Faris Khan, John Windle, Shane Tsai, Daniel Anderson, Gleb Haynatzki, and Niyada Naksuk
Nerve Growth Factor Receptor Kinase Inhibitor AG-879 Decreases Cell Viability in Glioblastoma Cell Lines
Blake M. Kidder, Raghupathy Vengoji Ph.D., Poonam Yadav, Surinder K. Batra Ph.D., and Nicole Shonka MD
Glioblastoma (GBM), a grade IV astrocytoma, is the most aggressive primary brain tumor and the mean adult survival time is a mere 14.6 months. Current treatment options include gross tumor resection, radiation treatment, and chemotherapy. Temozolomide is the standard drug utilized for chemotherapy, but it has a limited effectiveness, as patients commonly develop resistance after two and a half months. With no significant cures or treatments, it is pertinent to explore alternative avenues for disease mitigation. One possible pathway lies in the expression of Nerve Growth Factor Receptor (NGFR). Normally, NGFR is responsible for the differentiation of neurons, and it allows the central nervous system to respond to stimuli. If NGFR is dysregulated, cells begin to proliferate at extremely high rates. This uncontrolled proliferation has an association with the formation of GBM. Upregulation of NGFR in other cancers, specifically melanoma, has been associated with increased tumor resistance to cytotoxic T cells. When NGFR was inhibited in T cell resistant melanoma cell lines, T cell tumor killing levels were heightened. Our bioinformatic analysis using Gene Expression Profiling Interactive Analysis (GEPIA) revealed that NGFR expression significantly increased in GBM when compared to normal brain. RT-PCR analysis results display increased levels of NGFR transcripts in EPG (high grade mouse glioma cell line) when compared to lower grade gliomas. Moreover, our western blot analysis on human and mouse GBM cell lines showed increased expression of NGFR. Interestingly, NGFR inhibitor, AG-879 decreased the proliferation/viability of U251 GBM cells in a dose dependent manner.
Measuring Physiological Responses and Emotional Expression during Treatment of Pediatric Feeding Disorders: A Pilot Study
Margaret A. Kramer, Laura Elizabeth Phipps, James E. Gehringer, and Walker Arce
Our study aimed to identify possible associations between measures of physiological and behavioral data in children with feeding disorders during treatment. We found that all children demonstrated an average normal heart rate during the feeding appointments, all children demonstrated a primarily neutral emotion, and that three out of four participants had a brief elevation in heart rate that exceeded the normal range. In the future we would like to further investigate the possible effects of environmental factors at play (i.e., food, electronics) during meals that may correlate with elevations in heart rate.
Targeting Macrophages to Reduce Colorectal Cancer Metastasis: Diminished Effect in the Alcohol-injured Liver
Winnie Ladu, Ashley M. Mohr, Heather Richard, and Benita McVicker
Nicholas J. Lovick, Heather Jensen Smith, David G. Wagner, and Daphne Ly
Ductal carcinoma in situ (DCIS) is currently considered an early and localized form of ductal breast cancer stemming from the epithelial ductal cells. These lesions are largely heterogenous, categorized by their morphologies, amount of necrosis, and stromal changes. Even though 10-year mortality rate for DCIS is 1-2.6% while that of early invasive breast cancer is 7-10%. Yet, current recommended treatment for DCIS is breast-conserving surgery and radiation or mastectomy – the same treatment regimen recommended for early invasive breast cancer. This assumes that all DCIS will progress to invasive breast cancer if left untreated. However, mounting evidence indicates that a significant number of DCIS would remain indolence and never progress to invasive cancer. Current risk stratification is based on grade and hormone receptor (estrogen and progesterone) status. While the underlying mechanisms for DCIS to invasive cancer progression are not well understood, an improvement in the quantification of cellular morphology, the extracellular matrix and the metabolism modification of the tumor microenvironment could provide a more accurate and objective prognostication and treatment recommendations. DCIS is currently graded manually by a surgical pathologist using a representative number of areas on the slide. This risks grading bias between different pathologists. By using an automated software to measure quantifiable attributes such as nuclear density, size, and degree of variation of all areas of DCIS on the slide, we can have a more uniform and objective scoring system that would have minimal bias and variation. In addition, we will quantify heterogeneity of collagen arrangement, collagen fiber profile in the stroma as well as the metabolic modifications in the tumor microenvironment of “low risk” vs “high risk” DCIS to determine factors that could provide us with a better prognostication system.
Comparison Between Standard HABIT and HABIT-VR: Hand-Arm Function and Patient Acceptability Differences
Katelyn Malick, Toluwa Davies, Elizabeth Fortin, Kasey Mueller, Andrea Baraldi Cunha, Sandra L. Willett, and James E. Gehringer
Miguel Israel Martinez Nava, Melissa A. Leon, Sarah Maki, Tiffany Tanner, and Crystal Krause
Bariatric surgery is a long-term solution when dealing with obesity. The two most performed surgeries are sleeve gastrectomy (SG) and Roux-en-Y Gastric Bypass (RYGB). Both surgeries required a long-term follow-up with the patients as well as drastic changes in their lifestyle due to the alterations in their digestive system. For that reason, it is important to track health-related behaviors to keep a record of the patient’s health. In addition to a drastic change in the patient’s life, some of the health-related behaviors such as nocturnal eating and binge eating can lead to poorer surgical outcomes such as less weight loss. For that reason, there needs to be standardization around pre-health assessments in the bariatric world. The purpose of this literature review is to analyze preoperative surgery assessments for health behaviors including binge eating disorder (BED), nocturnal eating syndrome (NES), and alcohol use disorder (AUD).
Jack McEvoy, Elizabeth A. Kosmacek, Arpita Chatterjee, Molly Myers, Ariel Moats, Joshua McDowell, and Rebecca E. Oberley-Deegan
Prostate cancer is the second leading cause of cancer-related death in men. 10-20% of prostate cancer patients develop castration-resistant prostate cancer (CRPC) The median survival for CRPC patients is 14 months following diagnosis1. CRPC can spread to local tissues like the bladder, lymph nodes, and rectum but will also reach bone through distant metastasis. The metastasis of CRPC proves difficult for clinicians to treat. Redox active compounds can reduce growth and induce death of cancerous cells by the creation of ROS, which cannot be broken down by cancerous cells. This allows treatments like radiation and chemotherapy to be more effective in killing cancer.
Effect of MUC16 Blockade using the Humanized AR9.6 Antibody in Patient Derived Organoid Models of PDAC
Jordan N. Muirhead, Satish Sagar, Christabelle Rajesh, Adrian Black, and Prakash Radhakrishnan
Pancreatic Ductal Adenocarcinoma (PDAC) represents nearly 90% of all pancreatic cancer cases. 49,830 of the 62,210 patients diagnosed in 2022 are estimated to succumb to the malignancy. Early diagnosis of the disease is uncommon as most patients present with symptoms when the cancer is late-stage and metastatic. This decreases the likelihood of successful surgical resection and increases the dependency on standard of care chemotherapy leads to which is met with therapeutic resistance, demonstrated by the 5-year post-diagnosis survival rate of a mere 11.5%. Mucin-16, a heavy glycosylated transmembrane protein is overexpressed in more than 65% PDAC cases. AR9.6 is an anti-MUC16 antibody that has been recently humanized after evidence of its therapeutic potential was found in an orthotopic study utilizing the murine version of the antibody. The HuAR9.6 antibody efficiently binds MUC16 expressed on tumor cells, and can both inhibit downstream oncogenic signaling and elicit tumor killing by signaling the immune system to the tumor. In this project, we used RNA sequencing to evaluate the MUC16 mediated transcriptomic changes by using the humanized AR9.6 antibody in patient-derived organoid models of PDAC. To begin this study, organoids were developed using tumor cells from a primary PDAC tumor with a high MUC16 profile obtained from rapid autopsy patient #142 from the Rapid Autopsy Program at UNMC. The organoids were then treated in triplicates using a monoclonal antibody HuIgG as an isotype control due to its lack of specificity, and the test arm of study, HuAR9.6. These samples were treated with 40 ug/mL of the antibodies for a 24-hour period, post which RNA isolation was performed. RNA sequencing and subsequent Gene Ontology and KEGG enrichment analysis revealed a downregulation of genes involved in the Hippo signaling pathway, fat digestion and absorption and TGF-β signaling. Based on this gene expression profiling, we hypothesize that HuAR9.6 can slow tumor progression by downregulating the Hippo and TGF-β signaling pathways. In the future, we aim to robustly validate these results at the level of the proteome and assess if these results can be reproduced in multiple patient samples with the hope to translate this antibody to the clinic to be used in PDAC patients who have a high MUC16 expression.
Joshua Nguyen, Anupam Kotwal, Robert Bennett, Abbey Fingeret, Benjamin Swanson, Oleg Shats, Nicholas Whiteman, and Whitney Goldner
Thyroid cancer is the most common endocrine cancer, and its incidence has significantly increased over the last 40 years. Recent studies have suggested relaxin—a peptide hormone secreted by the ovaries during pregnancy with anti-fibrotic actions in chronic inflammation—as a potential marker of thyroid cancer occurrence and progression. However, it is unknown how relaxin behaves in benign versus malignant thyroid tissue. We hypothesized that relaxin levels would be decreased in benign and normal cancer-adjacent thyroid tissue relative to malignant tissue and increased in patients with lymphocytic thyroiditis, an autoimmune disorder involving chronic inflammation of the thyroid. Using the Integrated Cancer Repository for Cancer Research, benign, malignant, and normal cancer-adjacent thyroid tissue and accompanying clinical information were obtained. Tissue microarrays of each group were created, and immunofluorescence was performed to evaluate levels of relaxin. Our results indicated that relaxin is increased in malignant tissue relative to both normal cancer-adjacent and benign thyroid tissue, but there is no significant difference between benign and normal cancer-adjacent thyroid tissue. There is also an association between increased relaxin levels and lymphocytic thyroiditis. However, there is no association between significant differences in relaxin and other clinical factors like hypothyroidism and hyperthyroidism.
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