San Raffaele Telethon Institute for Gene Therapy

Safety of gene therapy and insertional mutagenesis

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Group Leader

Eugenio Montini

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Retroviral and Lentiviral Vectors allow the stable and long term expression of the therapeutic transgenes in therapeutically relevant cell types thanks to their ability to insert their genetic cargo in the genome of target cells. However, enhancer sequences and/or other genetic elements present in the integrated vector may interact with regulatory elements of the host cell genome and cause unwanted side effects.

The interest of our group are:

  • uncovering the interactions occurring between integrated viral vector and the surrounding host cell genome and understand how these interactions may affect the expression levels or the mRNA structure of cellular genes or induce changes in chromatin topology;
  • addressing the impact of vector insertions on the physiology of cells, tissues and, ultimately, of the whole organism.

To reach these aims, we have developed sensitive genetic mouse models for testing  the genotoxic potential of different vector types and backbone designs, state of the art high throughput retrieval and next generation sequencing of vector insertion sites and a solid bioinformatics and statistical framework to analyze and integrate genomic and phenotypic data.

Research activity

Currently we are studying the phenotype and fate of transplanted human and mouse hematopoietic stem and progenitor cells marked with specific vector constructs and evaluating their impact on the hematopoietic reconstitution in different mouse models.

Finally, we are using chromatin conformation capture technology to study the physical interactions between integrated lentiviral vector constructs and the surrounding chromatin. This approach provides the opportunity to observe and compare chromatin conformation changes occurring in the genome after the semi-random insertion of vector constructs containing different genetic elements.