Genome editing and hematopoietic stem cell based gene therapies
(PI: Mario Amendola)
Our laboratory is interested in developing safe and effective genome editing and classical gene therapy approaches to ex vivo modify hematopoietic stem cells (HSC) for the treatment of rare genetic disorders.
The current projects are mainly focused on:
- Correcting hemoglobinopathies, in particular sickle-cell disease and β-thalassemia, by exploiting artificial nucleases to repair the underlying disease causing mutations or to reactivate fetal globin expression.
- Developing new gene therapy approaches for the treatment of clotting disorders, in particular hemophilia A.
- Developing alternative ways to exploit HSC as therapeutic delivery vehicles, due to the ability of their progeny to extensively home to many tissues, including the central nervous system.
- Developing new genetic approaches to provide a fitness advantage to genetically corrected HSC over the affected ones.
PhD student, postdoc, bioinformatician position might be available in our group.
Non-dystrophic neuromuscular diseases
(PI: Ana Buj Bello)
This program focuses on the development and evaluation of novel strategies to correct genetic defects that affect the neuromuscular system. We are particularly interested in finding a cure for congenital neuromuscular diseases, such as myotubular myopathy. Our research concentrates on two main areas:
- AAV-mediated gene replacement therapy: the goal is to develop novel viral vectors, perform preclinical studies in animal models and clinical trials in patients.
- Genome editing: gene targeting at specific sites of the genome constitutes a promising approach for the in situ correction of a mutated gene and can be achieved by engineered nucleases such as the CRISPR/Cas9 system.
Molecular Immunology and Innovative Biotherapies
(PI: Anne Galy)
The laboratory is interested in the gene therapy of primary immune deficiencies and blood disorders. Blood and immune cells constitute a fascinating system to understand how gene therapy can correct human diseases at the molecular level and to study interactions with viral vectors and tolerance to genetic modifications.
- We propose the following research projects in the laboratory: We are currently developing a gene therapy for chronic granulomatous disease a severe immune deficiency with hyper-inflammation. A lentiviral vector is used to correct the NAPDH oxidase activity in phagocytes. We are interested in studying at the cellular level, the effects of oxidative metabolism and inflammation on hematopoiesis and on myeloid cell development, in the perspective of improving HSC engraftment and correction of this disease.
- We are using lentiviral vectors for gene transfer into HSC and are interested in improving the technology by engineering novel platforms enabling in vivo delivery and reiterated administration of vector through the development of immune invisible enveloped particles. A novel system developed in the laboratory will be characterized and used for gene transfer or gene editing applications in HSC.
The laboratory has access to state of the art molecular biology tools, is supplied by fresh cord blood cells, uses humanized mice, magnetic cell sorting, classical or spectral FACS and single cell analysis.
MicroRNA dysregulation in muscular dystrophy
(PI: David Israeli)
This program focuses on various aspects of miRNA dysregulation in muscular dystrophies. miRNA in the serum and urine of animal models and affected humans are analyzed using deep sequencing technologies. Identified target miRNAs are further investigated to define their potential as biomarkers in the clinics and to monitor the efficacy outcomes in preclinical and clinical trials.
The fundamental molecular pathophysiological mechanisms of muscular dystrophies are also investigated starting from a database of dysregulated non-coding RNAs in skeletal and cardiac muscle. The resulting hypotheses are tested in the laboratory in vitro and in vivo.
(PI: O.W. Merten)
The laboratory is developing novel technologies in the field of vectorology and gene transfer of adeno-associated viral (rAAV) and lentiviral (LV) vectors, Généthon’s workhorses for gene transfer. In this context, two research projects are proposed:
- The large scale production of AAV vectors is performed by the baculovirus/insect cell system and though this expression system is well performing and scalable, improvements are still required. The aim of this PostDoc project is to optimize the baculovirus expression system with respect to AAV vector titers, vector quality as well as potency in fine for reducing vector dose per patient, augment production efficiency and thus reduce treatment costs. Related to this the low full to empty particle ratio is a particular issue. These optimization activities are based on studies to be conducted for getting a better understanding of the fundamental AAV biology in general and in context of the baculovirus system, in particular.
- In order to improve vector production efficiency the actual transfection based manufacturing process should be replaced by stable producer cell lines able to grow in serum-free media in suspension using bioreactors. By using transposon/transposase and RMCE technologies, optimizing lentiviral helper functions, copy number, etc. novel efficient producer cell lines will be established and rigorously assessed with respect to growth and vector production under serum-free suspension conditions in view of production of high titer viral vectors of improved quality. The final aim is the implement such producer cell lines for routine production of vectors for clinical purposes.
Immunology and Liver Gene Transfer
(PI: Federico Mingozzi)
The research activities in our laboratory are mostly focused on the development of gene therapies based on adeno-associated virus (AAV) vectors and on the study of immune responses in gene transfer.
- Liver-directed gene transfer. The goal of this research, which involves laboratory investigations in vitro and in vivo, is to develop innovative gene therapies targeting hepatocytes to treat inherited diseases. Currently, we are leading the efforts towards the initiation of a clinical trial of liver gene therapy for Crigler-Najjar syndrome; we are also conducting preclinical research on glycogen storage diseases and other rare disease indications.
- Interactions of AAV vectors with the host immune system. This work is aimed at gaining a better understanding of the mechanisms determining the balance between tolerance and immunity in AAV vector mediated gene transfer. To this end, we perform studies in animal models and analyze immune responses to AAV in healthy donors and in subjects enrolled in gene therapy trials. The ultimate goal of this research is to enhance the safety and efficacy of AAV vector-mediated gene transfer by avoiding or modulating immune responses triggered by vector administration.
Progressive muscular dystrophies
(PI: Isabelle Richard)
This program focuses on progressive muscular dystrophies, in particular Duchenne Muscular Dystrophy resulting from mutations in the dystrophin gene and the recessive forms of Limb Girdle Muscular Dystrophies (LGMD2). These latter myopathies are grouped together on the basis of the common predominant involvement of proximal limb muscles. We are particularly interested in LGMD2A due to mutations in the gene coding for calpain 3, a protease of the skeletal muscle, LGMD2B which is due to defects in the dysferlin gene, a protein involved in membrane repair and the sarcoglycans, transmembrane proteins forming a complex at the plasma membrane.
Our guiding principle is to study fundamental biological processes using these diseases as models and to develop innovative therapeutic strategies to treat them. Currently, the research areas include:
Understanding the pathophysiological pathways:
- Regulation of sarcomere function through proteolysis or through miRNA pathways
- Epigenetics regulation in skeletal muscle;
- Intracellular trafficking;
Therapeutic interventions :
- Pharmacological strategies targeting the endoplasmic reticulum quality control;
- AAV mediated gene transfer. (including the development of strategies overcoming challenging issues for transfer : i. e. large genes, immune response against the transgene, etc)
The investigation of these topics is based on a variety of approaches (molecular, cell biological and in vivo imaging analyses, miRNA profiling, the recently described CRISPR/Cas9 technology, etc.) and is using in vitro (including iPSC) and in vivo models (mouse and rat).
Within this framework, the post-doctoral position will be adapted to the expertise and proposition of the candidate.