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> Schmidt, Heidelberg - Glimm, Heidelberg - von Kalle, Heidelberg <
Abstract I
Investigator I
Publications
Development of sensitive and unbiased integration site analyses to comprehensively assess biosafety and efficiency of innovative vectors
Integration site (IS) analyses are indispensable for assessing vector biosafety and following the clonal fate of gene-modified cells in vivo. However, current analyses rely on a restriction digest that severely limit integration site retrieval and consequently harbor the risk of drawing false conclusions. To enhance the safety and efficiency of gene therapy we will model the genomic accessibility to viral and non-viral IS defining a priori optimal enzyme combinations for distinct vector - cell gene transfer settings. Non-restrictive (nr) variants of the LAM-PCR will be implemented to reach comprehensive and quantitative IS assays coupled with next generation sequencing technologies. We will perform vector biosafety testings on gene transfer systems, in particular focusing on artificial zinc finger nucleases and integration-deficient lentiviral vectors and determine frequency and location of 'off-target' insertions. We will further determine the influence of IS on the proliferation and differentiation capacity of long-term hematopoietic repopulating cells in vitro and in vivo allowing novel insights into the physiology and cellular origin of insertion site driven clonal dominance in clinical gene therapy.
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Manfred Schmidt, Ph.D.
Department of Translational Oncology, G100
National Center for Tumor Diseases (NCT) and
German Cancer Research Center (DKFZ)
Im Neuenheimer Feld 581
69120 Heidelberg
e-mail: manfred.schmidt[at]nct-heidelberg.de
www.nct-heidelberg.de
Hanno Glimm, Dr. med., Priv. Doz.
Department of Translational Oncology
National Center for Tumor Diseases (NCT) and
German Cancer Research Center (DKFZ)
Im Neuenheimer Feld 460
69120 Heidelberg
e-mail: hanna.glimm[at]nct-heidelberg.de
www.nct-heidelberg.de
Christof von Kalle, Prof. Dr. med.
Department of Translational Oncology
National Center for Tumor Diseases (NCT) and
German Cancer Research Center (DKFZ)
Im Neuenheimer Feld 460
69120 Heidelberg
e-mail: christof.kalle[at]nct-heidelberg.de
www.nct-heidelberg.de
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5 selected Publications related to the project
Paruzynski A, Arens A, Gabriel R, Bartholomae CC, Scholz S, Wang W, Wolf S, Glimm H, Schmidt M, von Kalle C. Genome-wide high-throughput integrome analyses by nrLAM-PCR and next-generation sequencing. Nat Protoc. 2010 Aug; 5(8):1379-95.
Stein S, Ott MG, Schultze-Strasser S, Jauch A, Burwinkel B, Kinner A, Schmidt M, Krämer A, Schwäble J, Glimm H, Koehl U, Preiss C, Ball C, Martin H, Göhring G, Schwarzwaelder K, Hofmann WK, Karakaya K, Tchatchou S, Yang R, Reinecke P, Kühlcke K, Schlegelberger B, Thrasher AJ, Hoelzer D, Seger R, von Kalle C, Grez M. Genomic instability and myelodysplasia with monosomy 7 consequent to EVI1 activation after gene therapy for chronic granulomatous disease. Nat Med. 2010; 16(2):198-204.
Gabriel R, Eckenberg R, Paruzynski A, Bartholomae CC, Nowrouzi A, Arens A, Howe SJ, Recchia A, Cattoglio C, Wang W, Faber K, Schwarzwaelder K, Kirsten R, Deichmann A, Ball CR, Balaggan KS, Yáñez-Muñoz RJ, Ali RR, Gaspar HB, Biasco L, Aiuti A, Cesana D, Montini E, Naldini L, Cohen-Haguenauer O, Mavilio F, Thrasher AJ, Glimm H, von Kalle C, Saurin W, Schmidt M. Comprehensive genomic access to vector integration in clinical gene therapy. Nat Med. 2009; 15(12):1431-6.
Cartier N, Hacein-Bey-Abina S, Bartholomae CC, Veres G, Schmidt M, Kutschera I, Vidaud M, Abel U, Dal-Cortivo L, Caccavelli L, Mahlaoui N, Kiermer V, Mittelstaedt D, Bellesme C, Lahlou N, Lefrère F, Blanche S, Audit M, Payen E, Leboulch P, l'Homme B, Bougnères P, von Kalle C, Fischer A, Cavazzana-Calvo M, Aubourg P. Hematopoietic stem cell gene therapy with a lentiviral vector in X-linked adrenoleukodystrophy. Science. 2009; 326(5954):818-23.
Schmidt M, Schwarzwaelder K, Bartholomae C, Zaoui K, Ball C, Pilz I, Braun S, Glimm H, von Kalle C. High-resolution insertion-site analysis by linear amplification-mediated PCR (LAM-PCR) Nature Methods. 2007; 4(12), 1051-1057.
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