Monitoring of pediatric CGD gene therapy patients

Completed Research Project

Title / Titel: Monitoring of pediatric CGD gene therapy patients

Summary / Zusammenfassung: Within the first successful clinical X-CGD gene therapy study initiated in 2005 two adult patients were enrolled (Ott et al. Nat Med. 12, 401-9 (2006)). For monitoring of these patients a variety of functional tests were set up in our laboratory analyzing transgene presence and function on different levels as well as changes within the CGD phenotype.
These assays include the transgene detection (FACS analysis of gp91phox), analysis of gp91phox – p22phox assembly within the NADPH oxidase (Cytophrome B reduced minus oxidized difference spectrum) the qualitative detection (NBT reduction, DHR 123 oxidation, Chemiluminescence assay) and the quantitative analysis (Cytochrome C reduction assay) of respiratory burst activity as well as the functional analysis of anti-microbial activity of granulocytes (E.coli killing assay, A.fumigatus killing assay) as a result of reconstitution of respiratory burst activity. Furthermore we could quantify the percentage of gene modified cells within peripheral blood granulocytes by NBT reduction assay, as well as in bone marrow stem/progenitor cells by a colony assay combined with PCR based transgene detection in individual colonies of the colony assay.
These assays were set up for monitoring of adult patients as enrolled in the first clinical gene therapy study. Next generation clinical gene therapy study is intended to start in 2008. Therefore we miniaturize the present assays to be prepared for the limited sample volume as expected for pediatric patients of the next study.

Publications / Publikationen: Ott MG, Schmidt M, Schwarzwaelder K, Stein S, Siler U, Koehl U, Glimm H, Kuhlcke K, Schilz A, Kunkel H, Naundorf S, Brinkmann A, Deichmann A, Fischer M, Ball C, Pilz I, Dunbar C, Du Y, Jenkins NA, Copeland NG, Luthi U, Hassan M, Thrasher AJ, Hoelzer D, von Kalle C, Seger R, Grez M. Correction of X-linked chronic granulomatous disease by gene therapy, augmented by insertional activation of MDS1-EVI1, PRDM16 or SETBP1. Nat Med. (2006) 12: 401-9
Stein S, Siler U, Ott MG, Seger R, Grez M. Gene therapy for chronic granulomatous disease. Curr Opin Mol Ther. (2006) 8: 415-22

Keywords / Suchbegriffe: CGD, gene therapy

Project Leadership and Contacts / Projektleitung und Kontakte

Dr. Ulrich Siler (Project Leader)	ulrich.siler@kispi.uzh.ch
Dr. Janine Reichenbach	Janine.Reichenbach@kispi.uzh.ch
Reinhard Seger

Funding Source(s) / Unterstützt durch: Others
Donation USB AG by order of a client

Duration of Project / Projektdauer: Jan 2007 to Mar 2008