Comprehensive genomic access to vector integration in clinical gene therapy

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Richard Gabriel - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Ralph Eckenberg - , Genomining (Autor:in)
  • Anna Paruzynski - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Cynthia C. Bartholomae - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Ali Nowrouzi - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Anne Arens - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Steven J. Howe - , University College London (Autor:in)
  • Alessandra Recchia - , University of Modena and Reggio Emilia (Autor:in)
  • Claudia Cattoglio - , Vita-Salute San Raffaele University (Autor:in)
  • Wei Wang - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Katrin Faber - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Kerstin Schwarzwaelder - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Romy Kirsten - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Annette Deichmann - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)
  • Claudia R. Ball - , Deutsches Krebsforschungszentrum, DKFZ Standort Dresden, Umweltmonitoring und Endokrinologie (FoG), Nationales Centrum für Tumorerkrankungen Dresden, Nationales Zentrum für Tumorerkrankungen (NCT) Heidelberg (Autor:in)
  • Kamaljit S. Balaggan - , University College London (Autor:in)
  • Rafael J. Yá̃ez-Mũoz - , Royal Holloway University of London (Autor:in)
  • Robin R. Ali - , University College London (Autor:in)
  • H. Bobby Gaspar - , Great Ormond Street Hospital for Children NHS Trust (Autor:in)
  • Luca Biasco - , Fondazione Telethon (Autor:in)
  • Alessandro Aiuti - , Fondazione Telethon, University of Rome Tor Vergata (Autor:in)
  • Daniela Cesana - , Fondazione Telethon (Autor:in)
  • Eugenio Montini - , Fondazione Telethon (Autor:in)
  • Luigi Naldini - , Fondazione Telethon (Autor:in)
  • Odile Cohen-Haguenauer - , École normale supérieure Paris-Saclay, Hôpital Saint-Louis AP-HP (Autor:in)
  • Fulvio Mavilio - , University of Modena and Reggio Emilia, Vita-Salute San Raffaele University (Autor:in)
  • Adrian J. Thrasher - , University College London, Great Ormond Street Hospital for Children NHS Trust (Autor:in)
  • Hanno Glimm - , Deutsches Krebsforschungszentrum, DKFZ Standort Dresden, Nationales Centrum für Tumorerkrankungen Dresden, Nationales Zentrum für Tumorerkrankungen (NCT) Heidelberg (Autor:in)
  • Christof Von Kalle - , Deutsches Krebsforschungszentrum (DKFZ), Cincinnati Children's Hospital Medical Center (Autor:in)
  • William Saurin - , Genomining (Autor:in)
  • Manfred Schmidt - , Deutsches Krebsforschungszentrum (DKFZ) (Autor:in)

Abstract

Retroviral vectors have induced subtle clonal skewing in many gene therapy patients and severe clonal proliferation and leukemia in some of them, emphasizing the need for comprehensive integration site analyses to assess the biosafety and genomic pharmacokinetics of vectors and clonal fate of gene-modified cells in vivo. Integration site analyses such as linear amplification-mediated PCR (LAM-PCR) require a restriction digest generating unevenly small fragments of the genome. Here we show that each restriction motif allows for identification of only a fraction of all genomic integrants, hampering the understanding and prediction of biological consequences after vector insertion. We developed a model to define genomic access to the viral integration site that provides optimal restriction motif combinations and minimizes the percentage of nonaccessible insertion loci. We introduce a new nonrestrictive LAM-PCR approach that has superior capabilities for comprehensive unbiased integration site retrieval in preclinical and clinical samples independent of restriction motifs and amplification inefficiency.

Details

OriginalspracheEnglisch
Seiten (von - bis)1431-1436
Seitenumfang6
FachzeitschriftNature medicine
Jahrgang15
Ausgabenummer12
PublikationsstatusVeröffentlicht - Dez. 2009
Peer-Review-StatusJa

Externe IDs

PubMed 19966782

Schlagworte