Angelina
Sala
The cohesin complex is a highly conserved protein complex that regulates gene expression through DNA looping, which brings distant enhancers and promoters closer together in three- dimensional space. The gene encoding a core subunit of the cohesin complex, SMC1A, is located on the X chromosome and escapes X chromosome inactivation, meaning that females have higher expression than males. We hypothesize that differing levels of SMC1A in males and females affects the function of the cohesin complex and results in sex differences in genome- wide expression. The goal of this project is to investigate how X chromosome copy number affects the expression level and localization of each protein in the cohesin complex. We utilize indirect immunofluorescence and microscopy to investigate the cohesin proteins in different skin fibroblast cell lines derived from human subjects with varying numbers of X chromosomes. The fluorescent images are quantified using CellProfiler and the data extracted is analyzed in GraphPad Prism. Our preliminary data shows that the nucleic and cytoplasmic concentrations of SMC1A decreased as the number of X chromosomes increased. This result was unexpected, and further research is needed including experiments with more cell lines and optimizing image segmentation using an antibody that labels the cell membrane. Aside from the potential role of the cohesin complex in sex differences, it is also important to study because mutations in the complex result in various disorders, such as Cornelia de Lange Syndrome (CdLS). Studying the expression and localization of the cohesin complex may help us better understand these disorders. Investigating the molecular mechanisms of SCD while exploring stakeholder perspectives
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Authors:
Angelina Sala
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Pediatric sudden cardiac death (SCD) is often linked to genetic conditions such as arrhythmogenic cardiomyopathy (ACM) and congenital heart disease (CHD). This project takes a multidisciplinary approach to investigate the molecular basis of SCD and the implications of predictive genetic testing in at-risk populations. A biology-focused approach identified a novel genetic mechanism for autosomal recessive ACM associated with loss-of-function variants in TAX1BP3. Using induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs), pharmacological inhibition of TRPV4 mitigated calcium leak and spark frequency-highlighting a potential therapeutic target for ACM. Additionally, this project investigated several genetic mechanisms of hypoplastic left heart syndrome (HLHS), a severe form of CHD, by evaluating levels of apoptosis and cell proliferation in iPSC-CMs. Preliminary findings suggest increased apoptosis and reduced cell proliferation may lead to the underdeveloped left ventricle seen in patients, however, overall findings warrant further investigation into other mechanisms. On the other hand, a global health approach was incorporated by establishing a Community Advisory Board (CAB) to explore the ethical and clinical challenges of genetic risk prediction in CHD. Thematic analysis from the pilot CAB discussion highlighted concerns around clinical decision-making, resource allocation, and psychosocial impacts-emphasizing the need for standardized guidelines to ensure equitable and responsible use of genetic information. Altogether, this research advances precision medicine in pediatric cardiology by bridging molecular mechanisms with ethical considerations in genetic risk disclosure.
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Duke University / 2025
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Co-authors:
Angelina Sala