Molecular Drivers and Biomarkers of Spastic Cerebral Palsy

• Olivia Casale, Applied Molecular Biology & Biotechnology, University of Delaware

• Mona Batish, Medical and Molecular Sciences, University of Delaware

Cerebral palsy (CP) is a set of motor disorders that arise in early childhood, causing symptoms such as poor coordination and balance, uncontrolled movements, stiff or weak muscles, and speech, vision, learning, or behavioral problems. Currently, the molecular mechanisms behind these changes are poorly understood, making early diagnosis challenging. Determining the biological mechanisms and potential biomarkers of CP are critical for timely intervention that can prevent the progression of impairments and improve quality of life. To address this issue, this research is studying the roles of the proteins ABCA5 and MYOC in spastic CP, which were previously identified as proteins of interest through preliminary proteomics research in skeletal muscle. ABCA5 is a membrane protein that transports cholesterol and MYOC is a glycoprotein that enables the differentiation of myoblasts into myotubes. The expression levels of these proteins were evaluated using qRT-PCR, ELISA, and immunofluorescence. To test the functions of these proteins in CP, ABCA5 and MYOC knockdowns were created utilizing shRNA lentivirus. Additionally, cholesterol assays were performed to determine the role of ABCA5, and a fusion index assay was conducted to evaluate the function of MYOC in spastic CP. ABCA5 mRNA and protein expression were found to be upregulated in CP compared to controls (CN), with total cholesterol and free cholesterol upregulated in CP as well. Conversely, MYOC mRNA and protein expression were downregulated in CP compared to controls, negatively affecting the differentiation of myoblasts into myotubes. Ultimately, this project could support the discovery of novel biomarkers and provide a better understanding of their contributions to the pathways that drive the symptoms of spastic CP.