By investigating the interaction between different types of nucleases (i.e. nicking versus cleaving), donor DNA structures and target chromatin environments, this thesis provides important insights... Show moreBy investigating the interaction between different types of nucleases (i.e. nicking versus cleaving), donor DNA structures and target chromatin environments, this thesis provides important insights into how to improve the three crucial parameters of genome editing: efficiency, specificity and fidelity. The work presented in this thesis expand the range of possibilities for high-fidelity genetic manipulation of human cells. Show less
Bronchopulmonary dysplasia is the most common complication when premature birth occurs at less than 28 weeks gestational age. The general aim of this thesis is to explore the therapeutic... Show moreBronchopulmonary dysplasia is the most common complication when premature birth occurs at less than 28 weeks gestational age. The general aim of this thesis is to explore the therapeutic potential of interventions in signaling pathways, involved in lung development and oxidative stress-induced lung injury, to prevent or attenuate BPD in a neonatal rat model, in which experimental BPD is induced by exposure to hyperoxia. The therapeutic potential of the targeting compounds of signaling pathways was investigated by studying their beneficial effects on contributing factors to severe experimental BPD pathology, including aberrant alveolar and vascular development, inflammation, fibrosis, coagulation, vascular remodeling, pulmonary arterial hypertension and right ventricular hypertrophy. We found ⑴angiotensin II type 2 receptor ligand PD123319 attenuates hyperoxia-induced lung and heart injury at a low dose in newborn rats; ⑵ Metformin attenuates hyperoxia-induced lung injury in neonatal rats by reducing the inflammatory response; ⑶ Deficiency or inhibition of lysophosphatidic acid receptor 1 protects against hyperoxia-induced lung injury in neonatal rats; ⑷Adult lysophosphatidic acid receptor 1-deficient rats with hyperoxia-induced neonatal chronic lung disease are protected against lipopolysaccharide-induced acute lung injury; ⑸ Bone morphogenetic protein 9 protects against neonatal hyperoxia-induced impairment of alveolarization and pulmonary inflammation. Show less
The GABAergic system has been implicated in the pathogenesis of various anxiety disorders. Pharmacological treatments, like benzodiazepines, have been proven to target the GABA(A) receptors and... Show moreThe GABAergic system has been implicated in the pathogenesis of various anxiety disorders. Pharmacological treatments, like benzodiazepines, have been proven to target the GABA(A) receptors and exert quick-onset anxiolytic effect in anxiety patients. However, the side effects of these non-selective GABA(A)ergic compounds, such as sedation, postural imbalance, or potential abuse, limit their use for clinical anxiolysis. Based on the understanding of benzodiazepines’ mechanism of action, the emergence of α2,3 subtype-selective GABA(A) modulator is expected to provide a novel pharmacological approach that alleviates anxiety symptoms but spares the common undesired side effects. Most of these compounds are still in early clinical development, in which stage proof-of-mechanism studies are usually performed in healthy volunteers. The findings from our studies consistently present a similar pattern in the pharmacodynamic effect profiles of the α2,3 subtype-selective GABA(A) modulators versus those of the non-selective full GABA(A) agonist, lorazepam. Future application of anxiogenic symptom provocation models that combine subjective measurements and/or neuroendocrine biomarker assays may provide further construct validity for clinical anxiolytic effects of α2,3 subtype-selective GABA(A) modulators. Also, such findings are expected to provide insights into the translation of preclinical pharmacological properties of α2,3-subtype-selective GABA(A)ergic compounds to clinical effects in anxiety patients through human pharmacology studies. Show less
