Raquel Fueyo

All Publications

• Gain and loss of function variants in EZH1 disrupt neurogenesis and cause dominant and recessive neurodevelopmental disorders. Nature communications Gracia-Diaz, C., Zhou, Y., Yang, Q., Maroofian, R., Espana-Bonilla, P., Lee, C. H., Zhang, S., Padilla, N., Fueyo, R., Waxman, E. A., Lei, S., Otrimski, G., Li, D., Sheppard, S. E., Mark, P., Harr, M. H., Hakonarson, H., Rodan, L., Jackson, A., Vasudevan, P., Powel, C., Mohammed, S., Maddirevula, S., Alzaidan, H., Faqeih, E. A., Efthymiou, S., Turchetti, V., Rahman, F., Maqbool, S., Salpietro, V., Ibrahim, S. H., di Rosa, G., Houlden, H., Alharbi, M. N., Al-Sannaa, N. A., Bauer, P., Zifarelli, G., Estaras, C., Hurst, A. C., Thompson, M. L., Chassevent, A., Smith-Hicks, C. L., de la Cruz, X., Holtz, A. M., Elloumi, H. Z., Hajianpour, M. J., Rieubland, C., Braun, D., Banka, S., French, D. L., Heller, E. A., Saade, M., Song, H., Ming, G. L., Alkuraya, F. S., Agrawal, P. B., Reinberg, D., Bhoj, E. J., Martínez-Balbás, M. A., Akizu, N. 2023; 14 (1): 4109

  Abstract

  Genetic variants in chromatin regulators are frequently found in neurodevelopmental disorders, but their effect in disease etiology is rarely determined. Here, we uncover and functionally define pathogenic variants in the chromatin modifier EZH1 as the cause of dominant and recessive neurodevelopmental disorders in 19 individuals. EZH1 encodes one of the two alternative histone H3 lysine 27 methyltransferases of the PRC2 complex. Unlike the other PRC2 subunits, which are involved in cancers and developmental syndromes, the implication of EZH1 in human development and disease is largely unknown. Using cellular and biochemical studies, we demonstrate that recessive variants impair EZH1 expression causing loss of function effects, while dominant variants are missense mutations that affect evolutionarily conserved aminoacids, likely impacting EZH1 structure or function. Accordingly, we found increased methyltransferase activity leading to gain of function of two EZH1 missense variants. Furthermore, we show that EZH1 is necessary and sufficient for differentiation of neural progenitor cells in the developing chick embryo neural tube. Finally, using human pluripotent stem cell-derived neural cultures and forebrain organoids, we demonstrate that EZH1 variants perturb cortical neuron differentiation. Overall, our work reveals a critical role of EZH1 in neurogenesis regulation and provides molecular diagnosis for previously undefined neurodevelopmental disorders.

  View details for DOI 10.1038/s41467-023-39645-5

  View details for PubMedID 37433783

  View details for PubMedCentralID PMC10336078

• JMJD3 intrinsically disordered region links the 3D-genome structure to TGFbeta-dependent transcription activation. Nature communications Vicioso-Mantis, M., Fueyo, R., Navarro, C., Cruz-Molina, S., van Ijcken, W. F., Rebollo, E., Rada-Iglesias, A., Martinez-Balbas, M. A. 2022; 13 (1): 3263

  Abstract

  Enhancers are key regulatory elements that govern gene expression programs in response to developmental signals. However, how multiple enhancers arrange in the 3D-space to control the activation of a specific promoter remains unclear. To address this question, we exploited our previously characterized TGFbeta-response model, the neural stem cells, focusing on a ~374kb locus where enhancers abound. Our 4C-seq experiments reveal that the TGFbeta pathway drives the assembly of an enhancer-cluster and precise gene activation. We discover that the TGFbeta pathway coactivator JMJD3 is essential to maintain these structures. Using live-cell imaging techniques, we demonstrate that an intrinsically disordered region contained in JMJD3 is involved in the formation of phase-separated biomolecular condensates, which are found in the enhancer-cluster. Overall, in this work we uncover novelfunctions for the coactivator JMJD3, and we shed light on the relationships between the 3D-conformation of the chromatin and the TGFbeta-driven response during mammalian neurogenesis.

  View details for DOI 10.1038/s41467-022-30614-y

  View details for PubMedID 35672304

• Roles of transposable elements in the regulation of mammalian transcription. Nature reviews. Molecular cell biology Fueyo, R., Judd, J., Feschotte, C., Wysocka, J. 2022

  Abstract

  Transposable elements (TEs) comprise about half of the mammalian genome. TEs often contain sequences capable of recruiting the host transcription machinery, which they use to express their own products and promote transposition. However, the regulatory sequences carried by TEs may affect host transcription long after the TEs have lost the ability to transpose. Recent advances in genome analysis and engineering have facilitated systematic interrogation of the regulatory activities of TEs. In this Review, we discuss diverse mechanisms by which TEs contribute to transcription regulation. Notably, TEs can donate enhancer and promoter sequences that influence the expression of host genes, modify 3D chromatin architecture and give rise to novel regulatory genes, including non-coding RNAs and transcription factors. We discuss how TEs spur regulatory evolution and facilitate the emergence of genetic novelties in mammalian physiology and development. By virtue of their repetitive and interspersed nature, TEs offer unique opportunities to dissect the effects of mutation and genomic context on the function and evolution of cis-regulatory elements. We argue that TE-centric studies hold the key to unlocking general principles of transcription regulation and evolution.

  View details for DOI 10.1038/s41580-022-00457-y

  View details for PubMedID 35228718

• E proteins sharpen neurogenesis by modulating proneural bHLH transcription factors' activity in an E-box-dependent manner. eLife Le Dréau, G., Escalona, R., Fueyo, R., Herrera, A., Martínez, J. D., Usieto, S., Menendez, A., Pons, S., Martinez-Balbas, M. A., Marti, E. 2018; 7

  Abstract

  Class II HLH proteins heterodimerize with class I HLH/E proteins to regulate transcription. Here, we show that E proteins sharpen neurogenesis by adjusting the neurogenic strength of the distinct proneural proteins. We find that inhibiting BMP signaling or its target ID2 in the chick embryo spinal cord, impairs the neuronal production from progenitors expressing ATOH1/ASCL1, but less severely that from progenitors expressing NEUROG1/2/PTF1a. We show this context-dependent response to result from the differential modulation of proneural proteins' activity by E proteins. E proteins synergize with proneural proteins when acting on CAGSTG motifs, thereby facilitating the activity of ASCL1/ATOH1 which preferentially bind to such motifs. Conversely, E proteins restrict the neurogenic strength of NEUROG1/2 by directly inhibiting their preferential binding to CADATG motifs. Since we find this mechanism to be conserved in corticogenesis, we propose this differential co-operation of E proteins with proneural proteins as a novel though general feature of their mechanism of action.

  View details for DOI 10.7554/eLife.37267

  View details for PubMedID 30095408

  View details for PubMedCentralID PMC6126921

• Lineage specific transcription factors and epigenetic regulators mediate TGFβ-dependent enhancer activation. Nucleic acids research Fueyo, R., Iacobucci, S., Pappa, S., Estarás, C., Lois, S., Vicioso-Mantis, M., Navarro, C., Cruz-Molina, S., Reyes, J. C., Rada-Iglesias, Á., de la Cruz, X., Martínez-Balbás, M. A. 2018; 46 (7): 3351-3365

  Abstract

  During neurogenesis, dynamic developmental cues, transcription factors and histone modifying enzymes regulate the gene expression programs by modulating the activity of neural-specific enhancers. How transient developmental signals coordinate transcription factor recruitment to enhancers and to which extent chromatin modifiers contribute to enhancer activity is starting to be uncovered. Here, we take advantage of neural stem cells as a model to unravel the mechanisms underlying neural enhancer activation in response to the TGFβ signaling. Genome-wide experiments demonstrate that the proneural factor ASCL1 assists SMAD3 in the binding to a subset of enhancers. Once located at the enhancers, SMAD3 recruits the histone demethylase JMJD3 and the remodeling factor CHD8, creating the appropriate chromatin landscape to allow enhancer transcription and posterior gene activation. Finally, to analyze the phenotypical traits owed to cis-regulatory regions, we use CRISPR-Cas9 technology to demonstrate that the TGFβ-responsive Neurog2 enhancer is essential for proper neuronal polarization.

  View details for DOI 10.1093/nar/gky093

  View details for PubMedID 29438503

  View details for PubMedCentralID PMC5909450

• Epigenetic mechanisms in health and disease: BCEC 2017. Epigenetics Carbonell, A., Fueyo, R., Izquierdo-Bouldstridge, A., Moreta, C., Jordan, A. 2018; 13 (3): 331-341

  Abstract

  The Barcelona Conference on Epigenetics and Cancer (BCEC) entitled "Epigenetic Mechanisms in Health and Disease" was held in Barcelona, October 26-26, 2017. The 2017 BCEC was the fifth and last edition of a series of annual conferences organized as a joint effort of five leading Barcelona research institutes together with B-Debate. This edition was organized by Albert Jordan from the Molecular Biology Institute of Barcelona (IBMB-CSIC) and Marcus Bushbeck from the Josep Carreras Leukaemia Research Institute (IJC). Jordi Bernués, Marian Martínez-Balbás, and Ferran Azorín were also part of the scientific committee. In 22 talks and 51 posters, researchers presented their latest results in the fields of histone variants, epigenetic regulation, and chromatin 3D organization to an audience of around 250 participants from 16 countries. This year, a broad number of talks focused on the epigenetic causes and possible related treatments of complex diseases such as cancer. Participants at the 2017 BCEC elegantly closed the series, discussing progress made in the field of epigenetics and highlighting its role in human health and disease.

  View details for DOI 10.1080/15592294.2018.1434391

  View details for PubMedID 29384431

  View details for PubMedCentralID PMC5997171

• MicroRNA-200, associated with metastatic breast cancer, promotes traits of mammary luminal progenitor cells. Oncotarget Sánchez-Cid, L., Pons, M., Lozano, J. J., Rubio, N., Guerra-Rebollo, M., Soriano, A., Paris-Coderch, L., Segura, M. F., Fueyo, R., Arguimbau, J., Zodda, E., Bermudo, R., Alonso, I., Caparrós, X., Cascante, M., Rafii, A., Kang, Y., Martínez-Balbás, M., Weiss, S. J., Blanco, J., Muñoz, M., Fernández, P. L., Thomson, T. M. 2017; 8 (48): 83384-83406

  Abstract

  MicroRNAs are critical regulators of gene networks in normal and abnormal biological processes. Focusing on invasive ductal breast cancer (IDC), we have found dysregulated expression in tumor samples of several microRNAs, including the miR-200 family, along progression from primary tumors to distant metastases, further reflected in higher blood levels of miR-200b and miR-7 in IDC patients with regional or distant metastases relative to patients with primary node-negative tumors. Forced expression of miR-200s in MCF10CA1h mammary cells induced an enhanced epithelial program, aldehyde dehydrogenase (ALDH) activity, mammosphere growth and ability to form branched tubuloalveolar structures while promoting orthotopic tumor growth and lung colonization in vivo. MiR-200s also induced the constitutive activation of the PI3K-Akt signaling through downregulation of PTEN, and the enhanced mammosphere growth and ALDH activity induced in MCF10CA1h cells by miR-200s required the activation of this signaling pathway. Interestingly, the morphology of tumors formed in vivo by cells expressing miR-200s was reminiscent of metaplastic breast cancer (MBC). Indeed, the epithelial components of MBC samples expressed significantly higher levels of miR-200s than their mesenchymal components and displayed a marker profile compatible with luminal progenitor cells. We propose that microRNAs of the miR-200 family promote traits of highly proliferative breast luminal progenitor cells, thereby exacerbating the growth and metastatic properties of transformed mammary epithelial cells.

  View details for DOI 10.18632/oncotarget.20698

  View details for PubMedID 29137351

  View details for PubMedCentralID PMC5663523

• PRC2 Facilitates the Regulatory Topology Required for Poised Enhancer Function during Pluripotent Stem Cell Differentiation. Cell stem cell Cruz-Molina, S., Respuela, P., Tebartz, C., Kolovos, P., Nikolic, M., Fueyo, R., van Ijcken, W. F., Grosveld, F., Frommolt, P., Bazzi, H., Rada-Iglesias, A. 2017; 20 (5): 689-705.e9

  Abstract

  Poised enhancers marked by H3K27me3 in pluripotent stem cells have been implicated in the establishment of somatic expression programs during embryonic stem cell (ESC) differentiation. However, the functional relevance and mechanism of action of poised enhancers remain unknown. Using CRISPR/Cas9 technology to engineer precise genetic deletions, we demonstrate that poised enhancers are necessary for the induction of major anterior neural regulators. Interestingly, circularized chromosome conformation capture sequencing (4C-seq) shows that poised enhancers already establish physical interactions with their target genes in ESCs in a polycomb repressive complex 2 (PRC2)-dependent manner. Loss of PRC2 does not activate poised enhancers or induce their putative target genes in undifferentiated ESCs; however, loss of PRC2 in differentiating ESCs severely and specifically compromises the induction of major anterior neural genes representing poised enhancer targets. Overall, our work illuminates an unexpected function for polycomb proteins in facilitating neural induction by endowing major anterior neural loci with a permissive regulatory topology.

  View details for DOI 10.1016/j.stem.2017.02.004

  View details for PubMedID 28285903

• The histone demethylase PHF8 is a molecular safeguard of the IFNγ response. Nucleic acids research Asensio-Juan, E., Fueyo, R., Pappa, S., Iacobucci, S., Badosa, C., Lois, S., Balada, M., Bosch-Presegué, L., Vaquero, A., Gutiérrez, S., Caelles, C., Gallego, C., de la Cruz, X., Martínez-Balbás, M. A. 2017; 45 (7): 3800-3811

  Abstract

  A precise immune response is essential for cellular homeostasis and animal survival. The paramount importance of its control is reflected by the fact that its non-specific activation leads to inflammatory events that ultimately contribute to the appearance of many chronic diseases. However, the molecular mechanisms preventing non-specific activation and allowing a quick response upon signal activation are not yet fully understood. In this paper we uncover a new function of PHF8 blocking signal independent activation of immune gene promoters. Affinity purifications coupled with mass spectrometry analysis identified SIN3A and HDAC1 corepressors as new PHF8 interacting partners. Further molecular analysis demonstrated that prior to interferon gamma (IFNγ) stimulation, PHF8 is bound to a subset of IFNγ-responsive promoters. Through the association with HDAC1 and SIN3A, PHF8 keeps the promoters in a silent state, maintaining low levels of H4K20me1. Upon IFNγ treatment, PHF8 is phosphorylated by ERK2 and evicted from the promoters, correlating with an increase in H4K20me1 and transcriptional activation. Our data strongly indicate that in addition to its well-characterized function as a coactivator, PHF8 safeguards transcription to allow an accurate immune response.

  View details for DOI 10.1093/nar/gkw1346

  View details for PubMedID 28100697

  View details for PubMedCentralID PMC5397186

• EZH2 regulates neuroepithelium structure and neuroblast proliferation by repressing p21. Open biology Akizu, N., García, M. A., Estarás, C., Fueyo, R., Badosa, C., de la Cruz, X., Martínez-Balbás, M. A. 2016; 6 (4): 150227

  Abstract

  The function of EZH2 as a transcription repressor is well characterized. However, its role during vertebrate development is still poorly understood, particularly in neurogenesis. Here, we uncover the role of EZH2 in controlling the integrity of the neural tube and allowing proper progenitor proliferation. We demonstrate that knocking down the EZH2 in chick embryo neural tubes unexpectedly disrupts the neuroepithelium (NE) structure, correlating with alteration of the Rho pathway, and reduces neural progenitor proliferation. Moreover, we use transcriptional profiling and functional assays to show that EZH2-mediated repression of p21(WAF1/CIP1) contributes to both processes. Accordingly, overexpression of cytoplasmic p21(WAF1/CIP1) induces NE structural alterations and p21(WAF1/CIP1) suppression rescues proliferation defects and partially compensates for the structural alterations and the Rho activity. Overall, our findings describe a new role of EZH2 in controlling the NE integrity in the neural tube to allow proper progenitor proliferation.

  View details for DOI 10.1098/rsob.150227

  View details for PubMedID 27248655

  View details for PubMedCentralID PMC4852452

• Jumonji family histone demethylases in neural development. Cell and tissue research Fueyo, R., García, M. A., Martínez-Balbás, M. A. 2015; 359 (1): 87-98

  Abstract

  Central nervous system (CNS) development is driven by coordinated actions of developmental signals and chromatin regulators that precisely regulate gene expression patterns. Histone methylation is a regulatory mechanism that controls transcriptional programs. In the last 10 years, several histone demethylases (HDM) have been identified as important players in neural development, and their implication in cell fate decisions is beginning to be recognized. Identification of the physiological roles of these enzymes and their molecular mechanisms of action will be necessary for completely understanding the process that ultimately generates different neural cells in the CNS. In this review, we provide an overview of the Jumonji family of HDMs involved in neurodevelopment, and we discuss their roles during neural fate establishment and neuronal differentiation.

  View details for DOI 10.1007/s00441-014-1924-7

  View details for PubMedID 24950624

• RNA polymerase II progression through H3K27me3-enriched gene bodies requires JMJD3 histone demethylase. Molecular biology of the cell Estarás, C., Fueyo, R., Akizu, N., Beltrán, S., Martínez-Balbás, M. A. 2013; 24 (3): 351-60

  Abstract

  JMJD3 H3K27me3 demethylase plays an important role in the transcriptional response to different signaling pathways; however, the mechanism by which it facilitates transcription has been unclear. Here we show that JMJD3 regulates transcription of transforming growth factor β (TGFβ)-responsive genes by promoting RNA polymerase II (RNAPII) progression along the gene bodies. Using chromatin immunoprecipitation followed by sequencing experiments, we show that, upon TGFβ treatment, JMJD3 and elongating RNAPII colocalize extensively along the intragenic regions of TGFβ target genes. According to these data, genome-wide analysis shows that JMJD3-dependent TGFβ target genes are enriched in H3K27me3 before TGFβ signaling pathway activation. Further molecular analyses demonstrate that JMJD3 demethylates H3K27me3 along the gene bodies, paving the way for the RNAPII progression. Overall these findings uncover the mechanism by which JMJD3 facilitates transcriptional activation.

  View details for DOI 10.1091/mbc.E12-07-0561

  View details for PubMedID 23243002

  View details for PubMedCentralID PMC3564542