Gene therapy using viral vector against anxiety

Anxiety is an emotion that encompasses anxiousness and nervousness which can develop into neurological disorders caused by chronic stressors. Anxiety is mainly mediated by the amygdala that positively regulates the HPA axis, resulting in elevated levels of stress hormones in a person. A heightened HPA axis can further promote neuronal growth and increase spine density in the amygdala, forming a vicious cycle. One of the growth factors that modulate this development is Brain-Derived Neurotrophic Factor (BDNF). In this study, it was hypothesized that over-expression of BDNF in the BLA can lead to anxiety disorders. To prove this, the aim of the study was to clone BDNF gene into AAV vector, allowing co-transfection with pRC-AAV and phelper-AAV plasmids into HEK293T cells to generate viral particles. This project has been optimising BDNF gene cloning and viral packaging protocols to allow the continuation of BDNF gene research. Although unsuccessful in gene cloning, solutions were provided to rectify possible issues in the protocol. Future studies such as BDNF gene silencing and anxiety-related behaviour studies will be needed to support the hypothesis that BDNF gene is a major contributor in causing anxiety.Bachelor of Science in Biological Science

Pluripotent stem cell-derived committed cardiac progenitors remuscularize damaged ischemic hearts and improve their function in pigs

Abstract Ischemic heart disease, which is often associated with irreversibly damaged heart muscle, is a major global health burden. Here, we report the potential of stem cell-derived committed cardiac progenitors (CCPs) have in regenerative cardiology. Human pluripotent embryonic stem cells were differentiated to CCPs on a laminin 521 + 221 matrix, characterized with bulk and single-cell RNA sequencing, and transplanted into infarcted pig hearts. CCPs differentiated for eleven days expressed a set of genes showing higher expression than cells differentiated for seven days. Functional heart studies revealed significant improvement in left ventricular ejection fraction at four and twelve weeks following transplantation. We also observed significant improvements in ventricular wall thickness and a reduction in infarction size after CCP transplantation (p-value < 0.05). Immunohistology analyses revealed in vivo maturation of the CCPs into cardiomyocytes (CM). We observed temporary episodes of ventricular tachyarrhythmia (VT) in four pigs and persistent VT in one pig, but the remaining five pigs exhibited normal sinus rhythm. Importantly, all pigs survived without the formation of any tumors or VT-related abnormalities. We conclude that pluripotent stem cell-derived CCPs constitute a promising possibility for myocardial infarction treatment and that they may positively impact regenerative cardiology