Gene transfer and Gene therapy
Gene transfer and Gene therapy
Following the initial work done by the laboratory scientists (performed in collaboration with chemists in Brest and Rennes) on synthetic non-viral vectors for the gene transfection and the arrival in Brest in 2004 of Pierre Lehn, who had himself a long standing expertise in vectorology recognized at the international level, the important research effort of the “Gene transfer and Gene therapy” group during the last years has aimed to strengthen and further enhance that research avenue, in particular via the setting up of novel academic collaborations and the strengthening of its technological applications and thus its societal impact.
Our research over the last years has thus first aimed at pursuing the development of novel cationic lipids of the “lipophosphoramidate” type in collaboration with chemists of UMR CNRS 6521 located next to us in Brest. We have thus studied the transfection properties of lipophosphoramidates characterized by novel headgroups: Dicationic (Mevel et al, Bioconjug Chemistry, 2007), multivalent of the spermine type (Lamarche et al, Bioconjug Chemistry, 2007), or bearing an imidazolium group (Mevel et al, Chem BioChem, 2008). In parallel, we also evaluated the lipothiophosphoramidate, highlighting the key role of the polar head group (Fraix et al, Org Biomol Chem, 2012; Fraix et al, Org Biomol Chem, 2013). More recently, we have also reported the transfection activity of vectors with an unconventional lipidic moiety consisting of two di-unsaturated linoleic acid chains (Le Gall et al, J Med Chem, 2010), two phytanyl chains (Lindberg et al, Biomaterials, 2013) or bearing two different lipid chains (Le Corre et al, Org Biomol Chem, 2014).
Obviously, we have also pursued our collaboration with chemists from ENSCR (Ecole Nationale Supérieure de Chimie de Rennes) in Rennes. This allowed us to explore the gene transfection abilities of archaeoplexes, highly original formulations incorporating archaelipid derivatives in order to control the fluidity/rigidity balance of the liposomal membranes (Rethore et al, Chem Comm, 2007). Such derivatives could be pegylated and equipped with targeting ligands (folic acid) for cell targeting purposes (Laine et al, Chem, Eur J, 2008; Mornet et al, IJMS, 2013).
The main features of these various “Bio-inspired” vectors were presented in two review articles (Montier et al, Current Gene Therapy, 2008; Berchel et al, Biochimie, 2012). Of note, all these studies generally focused on in vitro gene transfection into various cell types (especially airway epithelial cells) and into the airway epithelium of the mouse in vivo (in particular with bioluminescent imaging and general cytotoxicity assessment in vivo) with a view to the lung gene therapy for Cystic Fibrosis (CF). Very recently, we demonstrated that such arsonium-based lipophosphoramidates are able to transfect epithelial cells and to kill bacteria at the same time, which represents a major interest in CF gene therapy (Le Gall et al, Adv Health Mat, 2013).
On the other hand, relying on our broad non viral expertise, we chose to also perform some cancer gene therapy studies. Here, lipophosphoramidates were found to be efficient in a gene-based anti-melanoma immunotherapy approach involving the transfection of a melanoma-specific antigen into human dendritic cells (Le Gallo et al, J Gene Med, 2008). In parallel, we have also developed, in the nude mouse, a tumour model consisting in the grafting of human melanocytes genetically-engineered to express the luciferase reporter gene and in which in vivo bioluminescence imaging allows thus to follow the tumour growth and the potential effect of a given treatment. Here, some lipofection-based “Suicide-gene” strategies were found to delay the tumour growth (David et al., Int J Pharm, 2012; David et al, J Gene Med, 2012). Some extinction strategies based on siRNA are ongoing work (Resnier et al, Biomaterials, 2013). We have also developed a metastatic mouse model, modifying the expression pattern of the melanocytes. This model and our competences in imaging are also useful to evaluate other approaches based on the delivery of anticancer drugs (David et al, Mol Ther, 2013; Rivera-Rodriguez et al, Int J Pharm, 2013; Lollo et al, Eur J Pharm Biopharm, 2014)
Collaborations, main funding and industrial development
As planned, we have also started novel academic collaborations. Thus, in the context of Biogenouest, we have been involved in the first large federative research programme entitled “Synthetic Vectors”; in addition to the 3 aforementioned teams for Brittany, this network brought together two other teams from the Pays de la Loire region, one from INSERM U915 in Nantes and another one from U1066 in Angers. For example, collaborative work performed in that context showed the feasibility of passive tumour targeting by using DNA nanocapsules (Morille et al, Biomaterials, 2010).
Next, we have also started a collaboration with chemists from Grenoble (UMR CNRS 5063) which has already allowed us to explore the transfection activity of cationic lipids with a neamine headgroup (Le Gall et al., Bioconj Chem, 2009).
Finally, whereas our work in the field of cancer gene therapy is supported by the Ligue contre le Cancer and the Canceropole Grand Ouest, it should be stressed here that, in a context of a “strategic” long-term research programme launched by AFM (Association française contre les Myopathies, the French muscular dystrophy foundation) in 2009, Pierre Lehn is leading a consortium bringing together biologists and chemists belonging to 6 different laboratories (Inserm U1078 and UMR CNRS 6521 in Brest, ENSCR UMR CNRS 6226 in Rennes, Inserm U915 in Nantes, UPR CNRS 4301 in Orléans and UMR CNRS 8585 in Evry). The goal of the research programme entitled “DNA nanoparticles for skeletal muscle and airway epithelium in vivo gene therapy” is to develop non-viral vector formulations optimized for in vivo gene transfection into the muscle and the airway epithelium. In this context, we learnt with John Wolff (Madison, WI, USA) how to perform Hydrodynamic Limb Vein administration. Now, we are also performing some successful aerosolisations of our complexes, leading to a pulmonary expression. On that plan, we have been collaborating with Steve Hyde’s group in Oxford (UK). The CF Gene Therapy development is also supported by VLM (Vaincre la Mucoviscidose, the French Cystic Fibrosis foundation).
Finally, as regards the technological applications, our research resulted in several patent applications and our broad expertise in non-viral vectors led to the creation of a technological platform in Biogenouest, platform termed “SynNanoVect” (directed by Tristan Montier), which involves teams from 4 laboratories (U1078 Brest, UMR CNRS 6521 Brest, UMR CNRS 6226 Rennes and U991 Rennes), whose goals are to synthesize and distribute our original synthetic formulations but also to perform some in vivo imaging analyses or electroporation assays. It was awarded the IBiSA label in 2008 and was renewed in 2012. Since May 2013, SynNanoVect has been certified ISO 9001.
• Tristan MONTIER (PU-PH, UBO - CHRU Brest)
• Pierre LEHN (PU-PH émérite, UBO - CHRU Brest)
• Véronique LAURENT (Ingénieur - Grant Manager, INSERM)
• Tony LE GALL (Chercheur, INSERM)
• Yann SIBIRIL (Assistant ingénieur, UBO)
• Angélique MOTTAIS (Doctorante, UBO - Association Gaétan Saleun, ancien membre)
• Yann LE GUEN (Doctorant, ED SICMA)
• Frédérique D'ARBONNEAU (MCU-PH, UBO - CHRU Brest)
• Chloé HENRY (Oncopédiatre - CHRU Brest, ancien membre)
Rosy GHANEM (Pharmacienne, Doctorante)
Tanguy HAUTE (CDD sept-dec 2019)
• UMR CNRS 6521 - Equipe "Phosphore & Vectorisation" (Brest)
• UMR CNRS 6226 "Sciences Chimiques" - Ecole Nationale Supérieure de Chimie (Rennes)
• UMR CNRS 7610 "Chimie des Polymères" - UPMC (Ivry)
• UPR CNRS 4301 - Equipe "Transfert d'acides nucléiques par des systèmes" (Orléans)
• UMR INSERM 915 - IRTUN, l'institut du thorax (Nantes)
• UMR CNRS 5063 "Chimie Moléculaire" (Grenoble)
• UMR INSERM 991 "Foie, métabolismes et cancer" (Rennes)
• Institute of Genetics and Molecular Medicine, University of Edinburgh (UK)
• Nuffield Department of Clinical Laboratory Sciences, University of Oxford (UK)
• Gene Delivery and Gene Therapy Lab, La Trobe University (Australia)
• Temple University, School of Pharmacy, Philadelphia (USA)
Collaborations INNOVATIVE THERAPY IN CANCEROLOGY
• UMR CNRS 6521 - Equipe "Phosphore & Vectorisation" (Brest)
• UMR CNRS 6226 "Sciences Chimiques de Rennes" - Ecole Nationale Supérieure de Chimie de Rennes (Rennes)
• UMR INSERM 1066 "MIcro et Nanomédecines biomiméTiques" (Angers)
• UMR CNRS 6290 "Institut de Génétique et Développement de Rennes" (Rennes)
• UMR Inserm 921 "Nutrition, Croissance et Cancer" (Tours)
• UMR 5253 CNRS/UM2/ENSCM/UM1 - Institut Charles Gerhardt Montpellier (ICGM) (Montpellier)
• Laboratoire de Toxicologie, Faculté de Pharmacie, Université Libre de Bruxelles (Bruxelles)
Pr Tristan MONTIER
Faculté de médecine
et des sciences de la santé
Université de Bretagne
22 rue Camille Desmoulins
29238 BREST Cedex 3
Tél : + 33 (0)2 98 01 80 80
Fax : + 33 (0)2 98 01 82 29
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