Development and Neuropharmacology
Director: Alain PROCHIANTZ, Pr. Collège de France
Our research is centered on the physiological function of homeoprotein intercellular transfer, a novel signaling mechanism discovered by our group and that of Alain Joliot, also a CIRB member. This research has basic and translational aspects.
Homeogenetic Extension and Boundary Formation
We have proposed that homeoprotein transfer could participate through iterative auto-inductions in the phenomenon of homeogenetic extension (Prochiantz and Joliot, Nature Reviews MCB 4 : 814-818, 2003; Holcman et al., J. Theoretical Biol. 249 : 503-517, 2007; Brunet et al TINS 30 : 260-267, 2007). This hypothesis was verified for the eye anlagen formation (Lesaffre et al. Develop. Neurosci., 2, 2. 2007).
An additional issue is the formation of boundaries also discussed in the papers referenced above. The idea is that boundaries form were homeoproteins meet because homeoproteins on either side of a boundary have reciprocal inhibitory activities. The hypothesis is presently tested in the developing neural tube where several boundaries exist that correspond to an established homeoproteins combinatorial code.
Collaborative with Dr. Jean-Léon Thomas (ICM, Salpêtrière, Paris)
This figure illustrates the homeoprotein signaling hypothesis at the core of our research program. As illustrated,homeoproteins are secreted and internalized by abutting cells where they regulate transcription and translation. Our program consists in identifying physiological functions for this novel mode of signal transduction, to identify translation and transcription (including epigenetic) targets, to verify if pathologies of unknown etiology can be explained by a failure in homeoprotein signaling and to use this information in translational research. References: A. Prochiantz and A. Joliot (2003). Can transcription factors function as cell-cell signaling molecules ? Nature reviews, Molecular Cell Biology, 4, 814-818. A. Joliot & A. Prochiantz (2004). Transduction peptides, from technology to physiology. Nature Cell Biology, 6, 189-196.
Homeoproteins Engrailed1 and Engrailed2 (collectively En1/2) respectively attract and repel retinal ganglion cell (RGCs) axons (Brunet et al., Nature, 438 : 94-98, 2005). The latter in vitro guidance requires En1/2 internalization by the growth cones, followed by local mRNA translation. In vivo 5% of En1/2 is extra-cellular with low anterior/high posterior graded expression. Neutralizing extra-cellular En1/2 leads to the overshooting of temporal axons into posterior regions of the tectum (Wizenmann et al., Neuron, 64 : 355-366, 2009).
Through two distinct approaches (metabolic labeling followed by mass spectrometry and identification of mRNAS on polysomes), several En1/2 local translation targets were identified. Among them, the presence of several mitochondrial complex I proteins led us to speculate that ATP may have a role in En1/2-regulated axon guidance. This hypothesis has been verified and our present model is that En1/2-induced ATP synthesis and release, followed by ATP hydrolysis into Adenosine leads to Adenosine Receptor 1 activation. This activation is necessary to observe the interaction between En1/2 and EphrinA5 signaling pathways (Stettler et al., in preparation).
Collaborative with Pr. Christine Holt (Univ. Cambridge, UK), Wolfgang Wurst (Von Helmotz, Institute, Munich, Germany), Dr. Olivier Stettler (IBPC, Paris)
In the developing neural tube, homeoproteins Nkx2.2 and Pax6 are involved in the regulation of specification and/or differentiation of oligodendrocytes. These myelin-forming cells arise as immature oligodendrocyte precursors (OPCs) in a specialized ventral domain of the ventricular and disperse in successive ventral and latero-dorsal waves. In analogy with the work on axon guidance (Brunet et al., Nature, 438 : 94-98, 2005), the presence of extracellular Pax6 and Nkx2.2 in the neural tube at E6 suggested a possible non-cell autonomous role of these factors in OPC migration.
This hypothesis was tested by gain and loss of function experiments to either neutralize or enhance extracellular Pax6 activity. Loss of function was achieved through the expression of secreted antibodies designed to antagonize Pax6 transfer. Secreted Pax6 antibody expression at E3 results in a reduction in the number of migrating Olig2+OPCs at E6. In contrast, electroporation of the protein Pax6 with a signal peptide increases the number of migrating cells. Identical conclusions were reached by culturing open book neural tubes in the presence of Pax6 neutralizing antibodies or of purified Pax 6 protein. All in all, these findings suggest a paracrine effect of Pax6 on OPCs, specifically promoting their migration (Hatton, Di Lullo et al., in preparation).
Collaborative with Dr. Jean-Léon Thomas (ICM, Paris)
Otx2 homeoprotein internalization by parvalbumin (PV) GABAergic neurons in layers III/IV of binocular visual cortex regulates the opening and closure of the critical period for binocular vision (Sugiyama et al., Cell, 134 : 508-520, 2008). This work is presently pursued along the following lines. We have identified the region of Otx2 which interacts with specific glycosaminoglycans expressed at the surface of PV cells and accounts for the specific capture of Otx2 by these neurons. The peptide corresponding to this recognition domain was synthesized and infused in the adult visual cortex leading to a blockade of endogenous Otx2 transfer and to a reopening of the critical period (Beurdeley et al, submitted).
As Otx2 is not synthesized in the cortex, its origin is an important issue. We are presently testing three non-exclusive possibilities: the retina, the choroid plexus and the pineal gland. Finally we have initiated a search for Otx2 cofactors and transcription/translation targets in PV cells.
Collaborative with Pr. Takao Hensch (Harvard University, Cambridge, USA) and Dr. Antonio Simeone (CEINGE, Naples, Italy)
Retinal ganglion cells (RGCs) are the projection neurons from the eye to the brain and their loss results in visual impairment in a number of diseases. Otx2 is a homeoprotein transcription factor expressed in the adult retina that transfers between bipolar cells and RGCs in the retina and can thus be taken up by RGCs. We find that Otx2 promotes the survival of axotomized adult RGCs in vitro, protects RGCs against NMDA-induced toxicity in vivo and preserves visual acuity in the latter model of neurotoxicity (Torero-Ibad, Rheey et al., submitted).
Collaborative with Dr. Serge Picaud (Quinze-vingts, Paris) and Fovea-SA.
Engrailed is expressed in the dopaminergic (DA) neurons of the adult Substantia Nigra (SN) and Ventral Tegmental Area (VTA). In a mouse heterozygote for Engrailed1 (En1LacZ mouse), DA neurons undergo progressive cell death in the adult. Moreover Engrailed infused in the SN/VTA and internalized by neurons in this region has a strong mDA pro-survival effect (Sonnier et al., J. Neurosci., 27 : 1063-1071, 2007). This suggests that Engrailed is in the Parkinson’s disease pathway and could be used both as therapeutic proteins (in the mouse) and as a mean to identify new therapeutic targets. The latter hypothesis is supported by new data showing that Engrailed protects against MPP-induced DA cell death in vitro and in vivo. This protection against MPP, but not 3-NP strongly suggests an implication of complex I (Alvarez-Sanchez, Fuchs et al., submitted).
A high throughput approach has allowed us to identify a series of transcription/translation targets and we are presently verifying them before testing their roles in gain and loss of function approaches.
Collaborative with Dr. A. Hartmann (Salpêtrière, Paris); Pr. Krebs (Ste Anne Hospital, Paris)
Selected Publication 2005-2018
- Di Nardo, A.A., Fuchs, J., Joshi, R.L., Moya, K.L. & Prochiantz, A. (2018). The Physiology of Homeoprotein Transduction. Physiol. Rev. 98, 1943–1982.
- Blaudin de Thé, F.-X., Rekaik, H., Peze-Heidsieck, E., Massiani-Beaudoin, O., Joshi, R.L., Fuchs, J. & Prochiantz, A. (2018). Engrailed homeoprotein blocks degeneration in adult dopaminergic neurons through LINE-1 repression. EMBO J.
- Apulei, J., Kim, N., Testa, D., Ribot, J., Morizet, D., Bernard, C., Jourdren, L., Blugeon, C., Di Nardo, A.A. & Prochiantz, A. (2018), Non-cell Autonomous OTX2 Homeoprotein Regulates Visual Cortex Plasticity Through Gadd45b/g. Cereb. Cortex.
- Prochiantz, A. (2018), How to navigate counter dogmatic research findings. EMBO J.
- Lee, H.H.C., Bernard, C., Ye, Z., Acampora, D., Simeone, A., Prochiantz, A., Di Nardo, A.A., & Hensch, T.K., (2017), Genetic Otx2 mis-localization delays critical period plasticity across brain regions. Mol. Psychiatry 22, 680–688.
- Bernard, C., Vincent, C., Testa, D., Bertini, E., Ribot, J., Di Nardo, A.A., Volovitch M., & Prochiantz, A. (2016), A Mouse Model for Conditional Secretion of Specific Single-Chain Antibodies Provides Genetic Evidence for Regulation of Cortical Plasticity by a Non-cell Autonomous Homeoprotein Transcription Factor. PLoS Genet. 12, e1006035.
- Bernard, C. & Prochiantz, A. (2016), Otx2-PNN Interaction to Regulate Cortical Plasticity. Neural Plast. 2016, 7931693.
- Kim, H.-T., Prochiantz, A. & Kim, J.W., (2016), Donating Otx2 to support neighbor neuron survival. BMB Rep 49, 69–70.
- Rekaik H., Blaudin de Thé F.-X., Prochiantz A., Fuchs J. & Joshi R.L. (2015), Dissecting the role of Engrailed in adult dopaminergic neurons - Insights into Parkinson disease pathogenesis. FEBS Lett. 589, 3786–3794.
- Prochiantz, A., 2015. [Investing in knowledge]. Med Sci (Paris) 31, 819–820.
- Rekaik H., Blaudin de Thé F.-X., Fuchs J., Massiani-Beaudoin O., Prochiantz A. & Joshi R.L. (2015), Engrailed Homeoprotein Protects Mesencephalic Dopaminergic Neurons from Oxidative Stress. Cell Rep. 13, 242–250.
- Dupont E., Prochiant, A. & Joliot A. (2015), Penetratin Story: An Overview. Methods Mol. Biol. 1324, 29–37.
- Quiñinao C., Prochiantz A. & Touboul J. (2015), Local homeoprotein diffusion can stabilize boundaries generated by graded positional cues. Development 142, 1860–1868.
- Huettl R.-E., Luxenhofer G., Bianchi E., Haupt C., Joshi R., Prochiantz A. & Huber A.B. (2015), Engrailed 1 Mediates Correct Formation of Limb Innervation through Two Distinct Mechanisms. PLoS ONE 10, e0118505.
- Nordström U., Beauvais G., Ghosh A., Pulikkaparambil Sasidharan B.C., Lundblad M., Fuchs J., Joshi R.L., Lipton J.W., Roholt A., Medicetty S., Feinstein T.N., Steiner J.A., Escobar Galvis M.L., Prochiantz A. & Brundin P. (2015), Progressive nigrostriatal terminal dysfunction and degeneration in the engrailed1 heterozygous mouse model of Parkinson’s disease. Neurobiol. Dis. 73, 70–82.
- Prochiantz A. & DiNardo A.A. (2015), Homeoprotein Signaling in the Developing and Adult Nervous System. Neuron 85, 911–925.
- Kim N., Acampora D., Dingli F., Loew D., Simeone A., Prochiantz A. & Di Nardo A. (2014), Immunoprecipitation and mass spectrometry identify non-cell autonomous Otx2 homeoprotein in the granular and supragranular layers of mouse visual cortex. F1000Res 3, 178.
- Bernard C., Kim H.-T., Torero Ibad R., Jung Lee E., Simonutti M., Picaud S., Acampora D., Simeone A., Ariel A. Di Nardo, Alain Prochiantz, Moya K.L.∗ & Woo Kim J.∗ (2014), Graded Otx2 activities demonstrate dose-sensitive eye and retina phenotypes, Human Molecular Genetics, Vol. 23, No. 7, 1742–1753.
- Golding B., Pouchelon G., Bellone C., Murthy S., Di Nardo A.A., Govindan S., Ogawa M., Shimogori T., Lüscher C., Dayer A., & Jabaudon D. (2014), Retinal Input Directs the Recruitment of Inhibitory Interneurons into Thalamic Visual Circuits, Neuron,81, 1057-1069, March 5.
- Spatazza J., Lee H.H., Di Nardo A.A., Tibaldi L., Joliot A., Hensch T.K. & Prochiantz A. (2013), Choroid-Plexus-Derived Otx2 Homeoprotein Constrains Adult Cortical Plasticity. Cell Report, Volume 3, Issue 6,1815-1823.
- Despras G., Bernard C., Perrot A., Cattiaux L., Prochiantz A., Lortat-Jacob H. & Mallet J.-M. (2013), Toward libraries of biotinylated chondroitin sulfate analogues: from synthesis to in vivo studies. Chemistry 19, 531-540.
- Spatazza J., Di Lullo E., Joliot A., Dupont E., Moya K.L. & Prochiantz A. (2013), Homeoprotein signaling in development, health, and disease: a shaking of dogmas offers challenges and promises from bench to bed. Pharmacol Rev. Jan 8;65(1):90-104.
- Beurdeley M., Spatazza J., Lee H., Sugiyama S., Bernard C., Di Nardo A., Hensch T. & Prochiantz A. (2012), Otx2 Binding to Perineuronal Nets Persistently Regulates Plasticity in the Mature Visual Cortex, The Journal of Neuroscience, July4, 2012-32(27): 9429-9437.
- Stettler O., Joshi R.L., Wizenmann A., Reingruber J., Holcman D., Bouillot C., Castagner F., Prochiantz A. & Moya K.L. (2012), Engrailed homeoprotein recruits the adenosine A1 receptor to potentiate ephrin A5 function in retinal growth cones. Development. Jan; 139(1):215-24.
- Layalle S., Volovitch M., Mugat B., Bonneaud N., Parmentier M.L., Prochiantz A., Joliot A. & Maschat F. (2011), Engrailed homeoprotein acts as a signaling molecule in the developing fly. Development, Jun; 138(11): 2315-23.
- Di Lullo E., Haton C., Le Poupon C., Volovitch M., Joliot A., Thomas J.-L. & Prochiantz A. (2011), Paracrine Pax6 activity regulates oligodendrocyte precursor cell migration in the chick embryonic neural tube. Development 138, 4991-5001.
- Torero Ibad R., Rheey J., Mrejen S., Forster V., Picaud S., Prochiantz A. & Moya K.L. (2011), Otx2 promotes the survival of damaged adult retinal ganglion cells and protects against excitotoxic loss of visual acuity in vivo. J. Neurosci., Apr 6;31(14):5495-503.
- Alvarez-Fischer D., Fuchs J., Castagner F., Stettler O., Massiani-Beaudoin O., Moya K.L., Bouillot C., Oertel W.H., Lombès A., Faigle W., Joshi R.L., Hartmann A. & Prochiantz A. (2011), Engrailed protects mouse midbrain dopaminergic neurons against mitochondrial complex I insults. Nat.Neurosci., Sep 14: 1260-66.
- Wizenmann A., Brunet I., Lam J., Sonnier L., Beurdeley M., Zarbalis K., Weisenhorn-Vogt D., Weinl C., Dwivedy A., Joliot A., Wurst W., Holt* C. & Prochiantz* A. (2009), Extracellular Engrailed participates in the topographic guidance of retinal axons in vivo. Neuron, 64, 355-366.
- De Toni A., Zbinden M., Epstein J.A., Ruiz I Altaba A., Prochiantz A. & Caillé I. (2008), Regulation of survival in adult hippocampal stem cell lineages by the homeodomain only protein HOP. Neural Dev, 3, 13.
- Kasatkin V., Prochiantz A. & Holcman D. (2008), Morphogenetic gradients and the stability of boundaries between neighbouring morphogenetic regions. Bulletin of Mathematical Biology, 70, 156-178.
- Sugiyama S., Di Nardo A., Aizawa S., Matsuo I., Volovitch M., Prochiantz* A. & Hensch* T.K. (2008), Experience-dependent transport of Otx2 homeoprotein in the visual pathway activates postnatal cortical plasticity. Cell, 134, 508-520.
- Brunet I., Di Nardo A., Sonnier L., Beurdeley M. & Prochiantz A. (2007), Shaping neural pathways with messenger homeoproteins. Trends in Neurosciences, 30, 260-267.
- Dupont E., Prochiantz A. & Joliot A. (2007), Identification of a signal peptide for unconventional secretion. J Biol Chem, 282, 8894-9000.
- Yves Agid, György Buzsáki, David M. Diamond, Richard Frackowiak, Jay Giedd, Jean-Antoine Girault, Anthony Grace, Jeremy J. Lambert, Husseini Manji, Helen Mayberg, Maurizio Popoli, Alain Prochiantz, Gal Richter-Levin, Peter Somogyi, Michael Spedding, Per Svenningsson & Daniel Weinberger (2007), How can drug discovery for psychiatric disorders be improved? Nature Reviews Drug Discovery, 6, 189-201.
- Sonnier L., Le Pen G., Hartman A., Bizot J.-C., Trovero F., Krebs M.-O.& Prochiantz A. (2007), Progressive loss of dopaminergic neurons in the ventral midbrain of adult mice heterozygote for Engrailed1: a new genetic model for neurological and psychiatric disorders. J Neurosci, 27, 1063-1071.
- Lesaffre B., Joliot A., Prochiantz A.& Volovitch M. (2007), Direct non-cell autonomous Pax6 activity regulates eye development in the zebrafish. Neural Development, 2, 2.
- Prochiantz A. (2007), A protein fusion a day keeps the aggregates away. Molecular Therapy, 15, 226-227.
- Prochiantz A. (2007), For protein transduction, chemistry can win over biology. Nat Methods, 4, 119-120.
- Holcman D., Kasatkin V. & Prochiantz A. (2007), Modeling homeoprotein intercellular transfer unveils a parsimonious mechaniss for gradient and boundary formation in early brain development. J Theor Biol, 249, 503-517.
- Di Nardo A., Nedelec S. (co-first), Trembleau A., Volovitch M., Prochiantz* A. & Montesinos M.L. (2007), Dendritic localization and activity-dependent translation of En1 homeodomain transcription factor mRNA. Mol Cell Neurosci, 35, 230-236.
- Linsik M.F., Christiaens B., Vandekerckhove J., Prochiantz A. & Rosseneu M. (2005), Penetratin-membrane association: W48/R52/W56 shield the peptide from the aqueous phase. Biophys J, 88, 1-14.
- Madeira A., Pommet J.M., Prochiantz A. & Allinquant B. (2005), SET protein (TAF1ß, I2PP2A) is involved in neuronal apoptosis induced by an Amyloid Precursor Protein cytoplasmic subdomain. FASEB J, 19, 1905-1907.
- Brunet I., Weinl C., Piper M., Trembleau A., Volovitch M., Harris B., Prochiantz* A., & Holt* C. (2005), Engrailed-2 guides retinal axons. Nature, 438, 94-98.
Prochiantz Alain, Professor Collège de France
Di Nardo Ariel, CR1 CNRS
Fuchs Julia, CR1 INSERM
Joshi Rajiv, DR2 CNRS
Moya Kenneth, DR2 CNRS
Volovitch Michel, Professor ENS Em.
Park Yang-ja, CDD
Postdoctoral fellows & PhD Students:
Kim Namsuk, Postdoctoral fellow
Kaddour Hadhemi, Postdoctoral fellow
Vargas Stéphanie, PhD student
Testa Damien, PhD student
Apulei Jessica, PhD student
Oliveira-Moreira Vanessa, PhD Student
Ravel-Godreuil Camille, PhD student
Peze-Heidsieck Eugenie, PhD student
Mazhar Bilal, PhD student
Leboeuf Melanie, PhD student
Beaudoin Olivia, IE1 CNRS
Torero-Ibad Raoul, IE2 CNRS
Dubreuil-Le Poupon Chantal, AI CDF
Cougnot Patricia, ADT CDF
Gibel-Russo Rachel, CDD AI CDF