SPP 1230:  Mechanisms of Gene Vector Entry and Persistence

• Subject Area: Medicine
• Term: from 2006 to 2018
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 18059742

Emerging gene-based therapies have shown surprising efficacy as well as severe side effects.
While efficacy was predicted by animal models, until recently the targeted investigation of dose-limiting side effects has been underrepresented in scientific activities. This situation has created major controversy in both scientific and regulatory communities.
Medicine and pharmacotherapy have always built their progress on the disciplines of pathology and toxicology. Likewise, side effects of genetic therapies reflect biological characteristics of the therapeutic moiety and the underlying disease and can only be prevented based on analytical insights into their mechanisms. An improved ability to predict the type, frequency and latency of side effects will automatically create a rational basis for the development and evaluation of innovative tools for somatic gene therapy. Because the topic of this emerging field is timely and requires an interactive and coordinated forum for unbiased evaluation and exploitation of its innovative potential, the Deutsche Forschungsgemeinschaft has installed a Priority Programme to investigate this topic. Three major (and interconnected) areas of vector-host interactions connect the members of this nationwide network:
(1) receptor and post-receptor restrictions to viral and non-viral gene delivery ("Cell Entry"),
(2) episomal persistence and chromosomal insertion of transgenes ("Nuclear Interaction"),
(3) cellular (differentiation and phylogeny) and milieu dependence of insertional mutagenesis and clonal kinetics of gene-modified cells ("Cell Fate").
The focus is on haematopoietic and lymphatic systems as these currently represent the most advanced targets in clinical studies. The Priority Programme combines expertise in basic and clinical haematology with molecular virology and cell biology. Bioinformatics is an integral component of this Priority Programme to model clonal competition following genetic interventions, and to coordinate multi-centre databases of gene vector insertion profiles and clonality of engineered cells. The Priority Programme thus creates a forum for focussed yet multidisciplinary approaches to unravel the mechanisms of transgene delivery and persistence. This approach results in essential insights into basic mechanisms of host-vector interactions and competitive clonal behaviour of (transgenic) cells in the organism. This will trigger competitive technology development and allow unbiased risk evaluation in basic research and clinical trials.

• DFG Programme: Priority Programmes

Projects

• Assessing and improving the safety profile of Sleeping Beauty transposon-mediated gene transfer in human cells (Applicant Ivics, Zoltan )
• Avoiding MDS/AML in genetic therapy for FA with complex retroviral vectors (Applicant Hanenberg, Helmut )
• Cell entry targeted gene transfer into quiescent cells of the hematopietic system by surface engineered lentiviral vectors (Applicant Buchholz, Christian )
• Characterization and Manipulation of Foamy Virus Vector Entry (Applicant Lindemann, Dirk )
• Control of epigenetic transgene silencing (Applicant Leonhardt, Heinrich )
• Coordinating central project of Priority Programme 1230 "Mechanisms of Gene Vector Entry and Persistence" (Applicant Baum, Christopher )
• Development of a non-viral episomal vector system enabling tissue-specific replication (Applicant Baiker, Armin )
• Development of sensitive and unbiased integration site analyses to comprehensivly assess biosafety and efficiency of innovative vectors (Applicant Schmidt, Ph.D., Manfred )
• Do retroviral vector insertions influence the fate of terminally differentiated cells such as T lymphocytes by insertional mutagenesis? (Applicant Fehse, Boris )
• Ein lentiviraler Vektor für die Transduktion von G0-arretierten Zellen. Analyse genetischer Determinanten (Applicant Cichutek, Klaus )
• Einfluss der T-Zell-Rezeptor-Diversität auf die Manifestation von T-Zell-Lymphomen/Leukämien im Mausmodel (Applicant Hansmann, Martin-Leo )
• Epigenetic of Vector-Chromatin Interactions (Applicant Grez, Manuel )
• Extrachromosomal plasmids and conditionally regulated gene vectors (Applicant Hammerschmidt, Wolfgang )
• Fate of plasmid DNA in host cells after bacteria-mediated gene transfer (Applicant Weiß, Siegfried )
• Generation of improved viral hybrid-vectors for stable transduction of mammalian cells (Applicant Ehrhardt, Ph.D., Anja )
• Humanized models to assess the genotoxicity of viral vectors in the context of hematopoietic stem cell expansion and in vivo selection (Applicant Moritz, Thomas )
• Improvement and application of a non-viral episome for mammalian cells (Applicant Lipps, Hans Joachim )
• Influence of nuclear architecture on gene vector persistence and expression (Applicant Dietzel, Steffen )
• Mathematical modeling of individual clone dynamics for genetically modified stem cells within the hematopoietic system (Applicant Röder, Ingo )
• Receptor-targeting and intracellular disassembly of gene vectors (Applicant Ogris, Manfred )
• Structure-function relationships of AAV capsids in post entry trafficking ans gene transfer (Applicant Kleinschmidt, Jürgen )
• Therapeutic gene targeting in human CD34+ cells. (Applicant Cathomen, Toni )
• Therapeutisches Potenzial und Risikoanalyse von T-Zellrezeptor genmodifizierten Knochenmark-Stammzellen für die adoptive T-Zelltherapie (Applicant Uckert, Wolfgang )
• Zelleintritt und intrazelluläre Prozessierung von rAAV targeting Vektoren und ihrer Genome im lympho-hämatopoetischen System (Applicant Büning, Hildegard )
• Zielzellen insertioneller Transformation in der Hämatopoese (Applicant Baum, Christopher )
• Zytokin-vermitteltes Targeting von Adenovirusvektoren zur effizienten Transduktion hematopoietischer Zellen (Applicant Kreppel, Florian )

• Spokesperson: Professor Dr. Christopher Baum