LABORATORIES
Neuronal generation and degeneration in vertebrates
Neuronal generation and degeneration in vertebrates
Research
Team
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Research
We are interested in the integration of the mechanisms of cell proliferation versus cycle exit and differentiation that result in the process of neurogenesis, as well as in the maintenance of the postmitotic state and neuronal homeostasis. We are also interested in the deregulation of these mechanisms and their association with neurodegenerative pathologies and aging.
Lines of investigation
1. Functional and pathological aspects of neuronal tetraploidy
The normal nervous system contains populations of tetraploid projection neurons in different tissues (Morillo et al., 2010; López-Sánchez et al., 2011; López-Sánchez and Frade, 2013). These neurons are functional (López-Sánchez and Frade, 2013). We are interested in defining the molecular mechanisms involved in its generation (Morillo et al., 2010; Morillo et al., 2012) and its subsequent maintenance (Ovejero-Benito and Frade, 2013; 2015), as well as its role in normal physiology, of the nervous system (Frade, 2010) and the possible epigenetic changes that can operate in them (Patiño-Parrado et al., 2017).
Specifically, we are interested in:
- Analyze the endogenous capacity of neurons to carry out the cell cycle (Frade and Ovejero-Benito, 2015; Walton et al., 2019).
- Characterize the molecular mechanisms that induce somatic tetraploidization in adult neurons, focusing on the role of the growth factor E2F4 and its phosphorylation by p38MAPK kinase (Morillo et al., 2012).
- Study the morphological and functional changes in neurons that undergo the neuronal tetraploidization process, including the compensatory mechanisms of gene expression in tetraploid neurons and possible epigenetic changes in these neurons.
- Characterize the pathophysiological effects of neuronal hyperploidization (Barrio-Alonso et al., 2018; 2020).
- Verify the participation of neuronal somatic tetraploidization in neurodegenerative diseases, focusing mainly on Alzheimer’s disease (López-Sánchez et al., 2017).
2. Role of the E2F4 transcription factor in cellular and tissue homeostasis in the nervous system: neurodegeneration and aging
E2F4 is a transcription factor traditionally known for its function in the regulation of quiescence in proliferating cells, which also plays a relevant role in the maintenance of cellular and tissue homeostasis (Ramón-Landreau et al., 2022). We work on the role of E2F4 phosphorylation at residues Thr248/Thr250 (Thr249/Thr251 in human E2F4) on the homeostatic capacity of this transcription factor, as well as the use of a mutant form of E2F4 incapable of being phosphorylated at these residues (E2F4DN) as a therapeutic multifactorial agent against Alzheimer’s disease and aging (López-Sánchez et al., 2021; 2022).
We are interested, therefore, in:
- To characterize the effects of E2F4 on different aspects of neuronal function, including transcription and translation, mitochondrial function, synaptic plasticity, proteostasis, oxidative stress, DNA damage, and neuronal survival.
- To understand the mechanism of action of E2F4DN as a therapeutic agent against neurodegeneration and aging.
3. Support to Tetraneuron for the development of a gene therapy based on the E2F4DN protein against Alzheimer’s disease
The biotechnology company Tetraneuron S.L., a spin-off of our laboratory that exploits our patent on the use of E2F4DN for the treatment of diseases caused by somatic tetraploidization, is currently conducting preclinical studies in rodents. Our laboratory gives scientific support in this work.
Most relevant methodology
To carry out these lines of research, a multidisciplinary approach is used based on histological, biochemical, molecular biology, cell biology and flow cytometry techniques, as well as in vivo protein expression using viral vectors, genetic modification in animal models, and behavioral studies. As experimental models, we use the mouse and human material from autopsies and donations.
Team
Jose María Frade
Principal Investigator
Noelia López Sánchez
Postdoctoral Resarcher
Christina Sánchez-Puelles
Postdoctoral Resarcher
Morgan Ramón Landreau
Predoctoral Researcher
María José Román
Laboratory technician
Publications
Five most relevant publications of the group
- López-Sánchez N, Garrido-García A, Ramón-Landreau M, Cano-Daganzo V, Frade JM (2021) E2F4-based gene therapy mitigates the phenotype of the Alzheimer’s disease mouse model 5xFAD. Neurotherapeutics 18: 2484-2503.
- Patiño-Parrado I, García-Gómez A, López-Sánchez N, Frade JM (2017) Strand-specific CpG hemimethylation, a novel epigenetic modification functional for gene silencing. Nucl. Acids Res. 45: 8822–8834.
- Formosa P, Ibañes M, Ares S, Frade JM (2012) Regulation of neuronal differentiation at the neurogenic wave front. Development 139: 2321-2329.
- Morillo SM, Escoll P, de la Hera A, Frade JM (2009) Somatic Tetraploidy in specific chick retinal ganglion cells induced by nerve growth factor. Proc. Natl. Acad. Sci. USA 107: 109-114.
- Frade JM (2000) Unscheduled cell cycle re-entry induced by NGF precedes cell death in nascent retinal neurones. J. Cell Sci.113: 1139-1148.
Publications of the last 10 years
- Sanz-García A, Sánchez Jiménez P, Granero-Cremades I, De Toledo M, Pulido P, Navas M, Fraga J, Frade JM, Pereboom D, Torres-Díaz CV, Ovejero-Benito MC (2022) Neuronal and astrocytic tetraploidy is increased in drug-resistant epilepsy. Appl. Neurobiol. 2022: e12873.
- Ramón-Landreau M, Sánchez-Puelles C, López-Sánchez N, Lozano-Ureña A, Llabrés-Mas AM y Frade JM (2022) E2F4DN transgenic mice: a tool for the evaluation of E2F4 as a therapeutic target in neuropathology and brain aging. Int. J. Mol. Sci. 23: 12093.
- López-Sánchez N, Ramón-Landreau M, Trujillo C, Garrido-García A y Frade JM (2022) A mutant variant of E2F4 triggers multifactorial therapeutic effects in 5xFAD mice. Mol. Neurobiol. 59: 3016-3039.
- Sánchez-Puelles C, Perea G y Frade JM (2021) E2F4DN-based gene therapy recovers long-term potentiation and hippocampal-dependent memory in homozygous 5xFAD mice. Alzheimers Dement. 17(Suppl.9): e057668.
- López-Sánchez N, Garrido-García A, Ramón-Landreau M, Cano-Daganzo V y Frade JM (2021) E2F4-based gene therapy mitigates the phenotype of the Alzheimer’s disease mouse model 5xFAD. Neurotherapeutics 18: 2484-2503.
- Frade JM (2021) 1920-2020 Instituto Cajal: cien años, cien logros/One hundred years, one hundred achievements. Ed. CSIC, Madrid. ISBN: 978-84-00-10863-2, eISBN: 978-84-00-10864-9
- García JM, Boscá L, Casado M, Bernad A, Álvarez-Dolado M, Villa R, Nacher E, Sánchez A, Cano D, Veiga E, Rodríguez F, Albiol FJ, Llosá G, Ratera I, Frade JM, Zapata M, Tapia N y Merino R (2021) Advanced Therapies (Bachiller D, Serrano, MC; Coordinators). In: CSIC Scientific Challenges: towards 2030. Vol. 4. Challenges in Biomedicine & Health (Topic Coordinators: Delgado M and Moros M). Editorial CSIC, Madrid. pág 157-175. ISBN: 978-84-00-10744-4
- Barrio-Alonso E, Fontana B, Valero M y Frade JM (2020) Pathological aspects of neuronal hyperploidization in Alzheimer’s disease evidenced by computer simulation. Front. Genet. 11:287.
- Walton CC, Zhang W, Patiño-Parrado I, Barrio-Alonso E, Garrido JJ y Frade JM (2019) Primary neurons can enter M-phase. Sci. Rep. 9: 4594. IF (2019): 3,998.
- Villoslada P, Vila G, Colafrancesco V, Moreno B, Fernandez-Diez B, Vazquez R, Pertsovskaya I, Zubizarreta I, Pulido-Valdeolivas I, Messeguer J, Vendrell-Navarro G, Frade JM, López-Sánchez N, Teixido M, Giralt E, Masso M, Dugas JC, Leonoudakis D, Lariosa-Willingham KD, Steinman L y Masseguer A. (2019) Axonal and myelin neuroprotection by the peptoid BN201 in brain inflammation. Neurotherapeutics 16: 808-827.
- Barrio-Alonso E, Hernández-Vivanco A, Walton, CC, Perea G y Frade JM (2018) Cell cycle reentry triggers hyperploidization and synaptic dysfunction followed by delayed cell death in differentiated cortical neurons. Sci. Rep. 8: 14316. IF (2018): 4,122.
- López-Sánchez N y Frade JM (2017) A mutant form of E2F4 prevents neuronal tetraploidization and cognitive deficits in 5xFAD mice without affecting A Alzheimers Dement. 13 (Supplement): P659–P661.
- Patiño-Parrado I, García-Gómez A, López-Sánchez N y Frade JM (2017) Strand-specific CpG hemimethylation, a novel epigenetic modification functional for gene silencing. Nucl. Acids Res. 45: 8822–8834.
- López-Sánchez N, Fontán-Lozano A, Pallé A, González-Álvarez V, Rábano A, Trejo JL y Frade JM (2017) Neuronal tetraploidization in the cerebral cortex correlates with reduced cognition in mice and precedes and recapitulates Alzheimer’s-associated neuropathology. Neurobiol. Aging 56: 50-66.
- Frade JM y López-Sánchez N (2017) Neuronal tetraploidy in Alzheimer and Aging. Aging 9: 2014-2015.
- Frade JM y Gage FH (2017) Genomic Mosaicism in neurons and other cell types. Springer Nature, New York. ISBN: 978-1-4939-7279-1 (eBook ISBN 978-1-4939-7280-7).
- López-Sánchez N, Patiño-Parrado I y Frade JM (2017) Quantification, isolation, and subsequent epigenetic analysis of tetraploid neurons by flow cytometry. In: Genomic Mosaicism in neurons and other cell types (Eds. Jose M. Frade and Fred H. Gage). Springer Nature, Nueva York, pág. 57-80.
- Slaninová I, López-Sánchez N, Šebrlová K, Vymazal O, Frade JM y Taborská E (2016) Introduction of Macarpine as a novel cell-permeant DNA dye for live cell imaging and flow cytometry sorting. Biol. Cell 108: 1-18.
- Ovejero-Benito MC y Frade JM (2015) p27Kip1 participates in the regulation of endoreduplication in differentiating chick retinal ganglion cells. Cell Cycle 14: 2311-2322.
- López-Sánchez N y Frade JM (2015) Flow cytometric analysis of DNA synthesis and apoptosis in the central nervous system using fresh cell nuclei. Methods Mol. Biol. 1254: 33-42.
- Frade JM y Ovejero-Benito MC (2015) Neuronal cell cycle: the neuron itself and its circumstances. Cell Cycle 14: 712-720.
- Galan A, Dergham P, Escoll P, de la Hera A, D’Onofrio PM, Magharious MM, Koeberle PD, Frade JM y Saragovi HU (2014) Neuronal injury external to the retina rapidly activates retinal glia, followed by elevation of markers for cell cycle re-entry and death in retinal ganglion cells. PLoS One 9: e101349.
- López-Sánchez N, Ovejero-Benito MC, Rodríguez-Ruiz C y Frade JM (2014) NGF/p75NTR in cell cycle and neuronal tetraploidy. En: Handbook of Neurotoxicity (Ed. Richard Kostrzewa). Springer Verlag, Heidelberg, pág. 1877-1897.
- López-Sánchez N y Frade JM (2013) Genetic evidence for p75NTR-dependent tetraploidy in cortical projection neurons from adult mice. J. Neurosci. 33: 7488-7500. IF (2013): 6,747.
- Ovejero-Benito MC y Frade JM (2013) Brain-derived neurotrophic factor-dependent cdk1 inhibition prevents G2/M progression in differentiating tetraploid neurons. PLoS One 5: e64890.
- López-Sánchez N y Frade JM (2013) Cell Cycle Analysis in the Vertebrate Brain Using Immunolabeled Fresh Cell Nuclei. Bio-protocol 3(22): e973. http://www.bio-protocol.org/e973
- Formosa-Jordan P, Ibañes M, Ares S y Frade JM (2013) Lateral inhibition and neurogenesis: novel aspects in motion. Int. J. Dev. Biol. 57: 341-350.
Contact
Where to find us
Laboratorio de generación y degeneración neuronal en vertebrados
Instituto Cajal CSIC. Avda. Doctor Arce, 37. 28002. Madrid
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Doctoral Thesis
- Doctoral student: Noelia López Sánchez
Title: Survival against trophic deprivation in neural tumor lines:
mitochondrial activation of JNK.
University: U.A.M. (Science Faculty).
Doctoral Program: Biochemistry, Molecular Biology and Biomedicine.
Date: May 3, 2006.
Qualification: Outstanding “cum laude”. - Doctoral student: Elsa Cisneros Child
Title: Regulation of the neurogenic capacity of vertebrate neural precursors during the cell cycle.
University: University of Alcalá (Faculty of Medicine).
Doctoral Program: Physiology.
Date: October 19, 2007.
Qualification: Outstanding “cum laude”. - Doctoral student: María del Carmen Ovejero Benito
Title: Regulation of tetraploidy maintenance in chicken retinal ganglion cells.
University: U.A.M. (School of Medicine).
Doctoral Program: Neuroscience.
Date: November 21, 2013.
Qualification: Outstanding “cum laude”. - Doctoral student: Iris Patiño Parrado
Title: Characterization of the methylation state in cytokines in tetraploid neurons of the murine cerebral cortex.
University: U.A.M. (Science Faculty).
Doctoral Program: Molecular Biosciences.
Date: January 15, 2018.
Qualification: Outstanding “cum laude”. - PhD Candidate: Chaska Walton Enríquez
Title: Mitotic biology of primary neurons.
University: U.A.M. (Science Faculty).
Doctoral Program: Molecular Biosciences.
Date: November 22, 2018.
Qualification: Outstanding “cum laude”. - Doctoral student: Estíbaliz Barrio Alonso
Title: Cell cycle reactivation in neurons and synaptic dysfunction: a new concept in Alzheimer’s disease.
University: U.A.M. (Science Faculty).
Doctoral Program: Molecular Biosciences.
Date: March 1, 2019.
Qualification: Outstanding “cum laude”. - Doctoral student: Morgan Ramón Landreau
Title: Modulation or the immune response by the neuronal expression of E2F4DN in the brain of the Alzheimer murine model 5xFAD.
University: U.A.M. (Science Faculty).
Doctoral Program: Molecular Biosciences.
Date: February 3, 2023.
Qualification: Outstanding “cum laude”. -
Patents
- Method to determine the risk of developing Alzheimer’s disease. Spanish Patent and Trademark Office (Patent No. 2 598 885; date: 10/02/2017). U.S. Patent No. US11047867B2. Grant date: 06/29/2021. Japan Patent No. JP6974184B2. Grant date: 12/01/2021. EU Patent EP3318874B1. Grant date: 09/09/2020. Operated by Tetraneuron, S.L (50% owner).
- Phosphorylation in residues Thr-248 and/or Thr-250 of the transcription factor E2F4 as a therapeutic target in pathological processes that occur due to somatic polyploidy. Spanish Patent and Trademark Office (Patent No. 2409779; date: 02/17/2014). U.S. Patent No. US9567384B2. Grant date: 02/14/2017. Japan Patent No. JP6100276B2. Grant date 01/27/2017. EU Patent No. EP2783696B1. Grant date: 08/09/2018. Operated by Tetraneuron, S.L
- Procedure for optimizing oligonucleotides and PCR amplification conditions for RACE (5′ and 3′-Rapid Amplification of cDNA Ends). Spanish Patent and Trademark Office (Patent No.: 200301290).
Spin-offs
- Tetraneuron, S.L. https://tetraneuron.com/es/
Collaborations
- Collaboration with Prof. Isabel Medina (Marine Research Institute, CSIC) in the regulation by E2F4DN of lipid and protein oxidation associated with aging.
- Collaboration with Prof. Antonino Cattaneo (European Brain Research Institute, Rome) and Prof. Xin-Fu Zhou (University of South Australia) on neuronal tetraploidy in mouse models of Alzheimer’s.
- Collaboration with Prof. Yves-A. Barde (Biozentrum, Basel) on signaling promoted by Trk/p75NTR.
Collaboration with Prof. Kazuaki Yoshikawa (Osaka University) function of MAGE proteins and regulation of the cell cycle mediated by p75NTR. - Collaboration with Dr. Dominique Morello (CNRS-Université Paul Sabatier, Toulouse) on the stabilization of mRNAs of neurogenic genes in neuronal precursors during mitosis.
- Collaboration with Dr. Antonio de la Hera (CSIC-Universidad de Alcalá). We collaborate in the analysis of neuronal tetraploidization.
- Collaboration with Dr. Miguel Ángel Cuadros (University of Granada) on the involvement of p75NTR in hereditary retinal dystrophies.
- Collaboration with Dr. Saúl Ares (Max-Planck-Institut für Physik komplexer Systeme) and Dr. Marta Ibáñez (University of Barcelona) on mathematical models of neurogenesis.
- Collaboration with Dr. Pablo Villoslada (IDIBAPS) in the analysis of specific p75NTR agonists.
Science dissemination
- Talk to High School and Baccalaureate students at I.E.S. Pradolongo, Madrid, March 15, 2023.
- Coordination of Baccalaureate students’ visit (I.E.S. Pradolongo) to the Cajal Institute. Madrid, March 13, 2023.
- Interview, “Hoy Por Hoy Madrid” Program, Cadena Ser Radio Station, January 17, 2023. https://cadenaser.com/cmadrid/2023/01/17/la-compania-madrilena-tetraneuron-desarrolla-una-nueva-terapia-genica-contra-el-alzheimer-radio-madrid/
- Frade JM (2022) New winds to tackle Alzheimer’s disease. Farmabiotec 5: 64-65. https://www.farmabiotec.com/revistas/numero-4-invierno-2022-2022
- Frade JM (2022) Gene therapy could be key to the cure for Alzheimer’s. elEconomista.es – Sanidad (https://revistas.eleconomista.es/sanidad/2022/septiembre/la-terapia-genica-podria-ser-clave-en-la-cura-del-alzheimer-MP11964109). Agosto 31, 2022.
- Frade JM (2022) Alzheimer and gene therapy. Gaceta Médica (https://gacetamedica. com/opinion/alzheimer-y-terapia-genica). July 8, 2022.
- Frade JM (2022) A necessary cure for Alzheimer’s. New Medical Economics (https://www.newmedicaleconomics.es/en-profundidad/una-necesaria-cura-para-el-alzheimer). May 11, 2022.
- “New advances in the treatment of neurodegenerative diseases. The role of gene therapy”, Round Table at Farmaforum 2022 (VIII Forum of the Biopharmaceutical Industry and Laboratory Technologies), Madrid, October 5, 2022.
- Interview, Dirigentesdigital.com, https://dirigentesdigital.com/economia/la-biotecnolo gia-espanola-frente-al-alzheimer, 25 agosto 2022.
- Interview, Farmabiotech (pages 26-28), https://www.farmabiotec.com/revistas/ numero-2-verano-2022.
- Interview, “Entre Probetas” Program, Radio 5, May 31, 2022. https://www.rtve.es/play/audios/entre-probetas/entre-probetas-comunidad-decidida-contra-alzheimer/6563568/
- Interview, “Emprende” Program, “RTVE 24 horas”, May 9, 2022.
- Coordinator of the “Hundred Tweets Cycle” of the Cajal Institute, Cien años, cien logros/One hundred years, one hundred achievements, 2020.
- President of the Organization Committee of the Centennial Events of the Cajal Institute, 2020.
- Interview on the Centennial of the Cajal Institute, “Dolphin Memory”, RNE Radio Station, July 18, 2020. https://www.rtve.es/radio/20200716/fado-alma-del-pueblo-portugues/2028901.shtml
- Participation in the Open Days of the Cajal Institute, February 21-23, 2020.
- “Personalized Medicine”, Round Table at the “Tomorrow Starts Today” Conference. IFEMA, Madrid, June 12, 2019.
- Frade JM (2016) Behind the origins of Alzheimer’s disease. Blog Areces Foundation. See at www.fundacionareces.es
- Frade JM (2013) DNA duplication in neurons: towards a new vision of Alzheimer’s disease.
- Conference in the context of the European Brain Month 2013. Toledo, May 14, 2013.
- Frade JM (2013) Nerve growth factor six decades later (Special Rita Levi-Montalcini). Spanish Society of Biochemistry and Molecular Biology. See at www.sebbm.es
- Frade JM (2012) Exhibition “Gallery of Portraits Women in Biochemistry” that includes the biographical profile of Rita Levi-Montalcini. Vice President for Research of the University of Seville, Brazilian Pavilion. September 17, 2012 – October 17, 2012.
- Frade JM (2012) Neuronal tetraploidy and Alzheimer’s. Round Table on Alzheimer’s Disease. Complutense University of Madrid. Brain Week. Madrid, March 21, 2012.
- Frade JM (2012) Biographical profile of Rita Levi-Montalcini. In the context of the role played by women scientists throughout the last century. Web page of the Spanish Society of Biochemistry and Molecular Biology. See at www.sebbm.es
- Frade JM (2011) Cellular Suicide as a Vital Necessity. Cycle of Conferences “Two Days of Science”. Cajastur and Science. Cajastur Cultural Center. Oviedo, April 25, 2011.
- Frade JM (2010) Sortilin and its role in signaling the p75NTR neurotrophin receptor.
- Implications in retinitis pigmentosa. VISION 35: 17-20.
Frade JM (2008) Retinitis pigmentosa and proNGF-mediated photoreceptor cell death. A new therapeutic pathway. VISION 33: 14-16. - Frade JM (2007) Experimental models of retinal dystrophies and apoptosis induced by the p75NTR receptor. VISION 31: 6-7.
Frade JM (2007) Zellen, hört die Signale! In: Gehirn&Geist, vol. 5/2007, p. 66-71. - Frade JM (2006) Knowledge of the degenerative mechanisms of photoreceptors will facilitate the design of therapeutic agents against retinal dystrophies. VISION 28: 10-11.
- Frade JM (2005) Neurotrophins and their receptors. In: MIND AND BRAIN, vol. 14, p. 10-15.
- Frade JM (2002) Follow the development of an embryo through a magnifying glass. In: II Madrid Science Fair, p. 195. Community of Madrid. SM Editions, Madrid.
Neuroscience Research Center dependent on the CSIC. Founded in 1920 and initially directed by Santiago Ramón y Cajal. World reference in the study of the brain. Custodian of the Cajal Legacy.
Activities