HUBERT DE VERNEUIL


Email: Hubert.De.Verneuil@u-bordeaux2.fr

Team: Biotherapies of genetic diseases and cancers

Website: www.u1035.u-bordeaux2.fr

Project 1:  iPSC and porphyrias

Erythropoietic porphyrias are inherited diseases with loss of enzymatic activities involved in heme biosynthesis. Enzymatic deficit leads to toxic accumulation of porphyrins in bone marrow, red blood cells, spleen and liver. Depending on the porphyria type, skin presents photosensitivity manisfestations with variable severity. The only curative treatment for severe porphyrias relies on allogenic bone marrow grafts. Our research team develops gene therapy for erythropoietic prophyria to transfer therapeutic genes with lentiviral vectors in autologous hematopoietic stem cells. We are currently focusing on mouse and human induced pluripotent stem cells (iPSC) obtained after reprogramming somatic cells. To treat erythropoietic porphyrias, we test whether it is possible to correct gene deficiencies in these cells before they are committed in hematopoietic and hepatic lineages.

Project 2: Roles of HIFs in hematopoietic cell lineage

Physiologic levels of oxygen (02) are strictly regulated in mammal tissues; oxygen levels vary between tissues and also within the same tissue. However, O2 implications in cell functions have long been ignored, even in cell models, which are usually cultured in vitro with O2 concentrations corresponding to atmospheric levels (21%, normoxia). Noticeably however, the most oxygenated tissues do not contain more than 14% O2 in the body. The Hypoxia Inducible Factors (HIFs) are major actors of O2 response and are key factors for the regulation of genes involved in cell survival, apoptosis, motility, cytoskeleton structure, adhesion and energetic metabolism of normal cells. They are also part of tumor cell growth and resistance to anti-cancer drugs. It is now well documented that hematopoietic cells are spread in the bone marrow along an O2 gradient: stem cells (HSC) are located in niches poor in oxygen (hypoxic even anoxic regions), which helps maintaining the undifferentiated state. Cells progressing into differentiation move towards vessels. We examine the impact of HIFs altered expression either by RNA interference or by lentiviral transfer of HIF cDNAs in human HSCs. We found that HIF downregulation in vitro leads to diminished levels of red cell lineage progenitors, with various amplitudes depending on the targeted factor. In vivo, long-term hematopoiesis was inhibited. To dissect molecular mechanisms involved in HIF functions during hematopoiesis, we seek for genes which regulation is impaired in HSCs after altered HIF expression.

Expertise

  1. Production of lentiviral and AAV vectors for the scientific community.
  2. Murine and human iPS cells.
  3. Mouse models of erythropoietic porphyrias

Main publications

  1. Bedel A, Taillepierre M, Guyonnet-Duperat V, Lippert E, Dubus P, Dabernat S, Mautuit T, Cardinaud B, Pain C, Rousseau B, Lalanne M, Ged C, Duchartre Y, Richard E, de Verneuil H, Moreau-Gaudry F. Metabolic Correction of Congenital Erythropoietic Porphyria with iPSCs Free of Reprogramming Factors.  Am J Hum Genet. 2012 Jul 13;91(1):109-21.
  2. To-Figueras J, Ducamp S, Clayton J, Badenas C, Delaby C, Ged C, Lyoumi S, Gouya L, de Verneuil H, Beaumont C, Ferreira GC, Deybach JC, Herrero C, Puy H. ALAS2 acts as a modifier gene in patients with congenital erythropoietic porphyria. Blood. 2011 Aug 11;118(6):1443-51.
  3. Duchartre Y, Petit N, Moya C, Lalanne M, Dubus P, Verneuil H, Moreau-Gaudry F, Richard E. Neonatal bone marrow transplantation prevents liver disease in a murine model of erythropoietic protoporphyria. J Hepatol. 2011 Jul;55(1):162-70.
  4. Robert-Richard E, Lalanne M, Lamrissi-Garcia I, Guyonnet-Duperat V, Richard E, Pitard V, Mazurier F, Moreau-Gaudry F, Ged C, de Verneuil H. Modeling of congenital erythropoietic porphyria by RNA interference: a new tool for preclinical gene therapy evaluation. J Gene Med. 2010 Aug;12(8):637-46.
  5. James C, Mazurier F, Dupont S, Chaligne R, Lamrissi-Garcia I, Tulliez M, Lippert E, Mahon FX, Pasquet JM, Etienne G, Delhommeau F, Giraudier S, Vainchenker W, de Verneuil H. The hematopoietic stem cell compartment of JAK2V617F-positive myeloproliferative disorders is a reflection of disease heterogeneity. Blood. 2008 Sep 15;112(6):2429-38.
  6.  Richard E, Robert-Richard E, Ged C, Moreau-Gaudry F, de Verneuil H. Erythropoietic porphyrias: animal models and update in gene-based therapies. Curr Gene Ther. 2008 Jun;8(3):176-86. Review.
  7. Robert-Richard E, Moreau-Gaudry F, Lalanne M, Lamrissi-Garcia I, Cario-André M, Guyonnet-Dupérat V, Taine L, Ged C, de Verneuil H. Effective gene therapy of mice with congenital erythropoietic porphyria is facilitated by a survival advantage of corrected erythroid cells. Am J Hum Genet. 2008 Jan;82(1):113-24.
  8. Robert-Richard E, Richard E, Malik P, Ged C, de Verneuil H, Moreau-Gaudry F. Murine retroviral but not human cellular promoters induce in vivo erythroid-specific deregulation that can be partially prevented by insulators. Mol Ther. 2007 Jan;15(1):173-82.
  9. Lyoumi S, Abitbol M, Andrieu V, Henin D, Robert E, Schmitt C, Gouya L, de Verneuil H, Deybach JC, Montagutelli X, Beaumont C, Puy H. Increased plasma transferrin, altered body iron distribution, and microcytic hypochromic anemia in ferrochelatase-deficient mice. Blood. 2007 Jan 15;109(2):811-8.
  10. Robert-Richard E, Ged C, Ortet J, Santarelli X, Lamrissi-Garcia I, de Verneuil H, Mazurier F. Human cell engraftment after busulfan or irradiation conditioning of NOD/SCID mice. Haematologica. 2006 Oct;91(10):1384.