| Nederlands

Pompe disease : towards gene correction using targeted nucleases

(2010) Geel, Tessa Marieke

Pompe disease is an inherited metabolic disorder caused by a mutation in the gene encoding the enzyme acid-α-glucosidase (GAA). In this dissertation, novel strategies were investigated to improve the treatment of Pompe disease. The only currently available therapy (Enzyme Replacement Therapy) is hampered by several disadvantages such as inefficient uptake of the enzyme. We demonstrated that a known efficient DNA carrier (SAINT) could also be used to effectively deliver proteins, including lysosomal enzymes and restriction enzymes. Moreover, SAINT delivered GAA siRNA to Pompe relevant cell types in an animal model. As this approach does not cure the underlying genetic defect in Pompe disease, we investigated whether gene correction (via homologous recombination) represents an alternative approach for Pompe disease. As the frequency of homologous recombination is very low in a cell, induction of a double strand break (DSB) is essential. Double strand breaks can be induced by restriction enzymes, which recognize 4-8 base pairs in the DNA. As these recognition sites are abundantly present in the genome, it is necessary to create GAA gene specificity by fusing DNA binding domains to the enzymes. We developed a read out system and tested GAA gene targeting agents for their efficiency in inducing DNA damage and/or homologous recombination. The studies described in this dissertation have made a contribution to the progress of two new strategies for Pompe disease: improved delivery of the GAA enzyme to the affected cells and targeted gene correction to obtain an actual cure for the severe life threatening disease.

	Titel en inhoud
	Hoofdstuk 1
	Hoofdstuk 2
	Hoofdstuk 3
	Hoofdstuk 4
	Hoofdstuk 5
	Hoofdstuk 6
	Hoofdstuik 7
	Hoofdstuk 8
	Hoofdstuk 9
	Nederlandse samenvatting
	Publikaties
	Dankwoord
	Appendix
	Volledige dissertatie