grupo

Group name: Otology & Neurotology- Genomics of vestibular disorders

    Group leader
  • Jose Antonio López Escámez
  • Senior Researcher
  • Juan Manuel Espinosa Sánchez
  • Miguel Ángel López Nevot
  • PHD Students
  • Pablo Roman-Naranjo Varela
  • Marisa Flook Pereira
  • Sana Amanat

Presentation Video

SCIENTIFIC INTEREST AREAS

Vertigo and imbalance are a major health problem caused by several disorders of the vestibular system involving the inner ear and the brain. The most common causes are benign paroxysmal positional vertigo, vestibular migraine and Menière’s disease. Among them, Menière’s disease (MD) is the most severe form of vertigo and tinnitus.

Meniere is a set of rare disorders of the inner ear causing sensorineural hearing loss, vertigo episodes and tinnitus with a prevalence of 75 cases/100.000 population in Spain. Familial Menière’s disease represents 8-9% of the cases and it has a genetic heterogeneity with an autosomal dominant inheritance pattern. There are several clinical subgroups, suggesting different mechanisms of disease including autoimmune, autoinflamatory and genetic factors. Our group has compiled the largest collection of human DNA samples from MD patients including >100 multicase families and > 2000 sporadic cases to define the genetic contribution associated with Md. For this purpose we have genotyped sporadic cases defining common variants in noncoding regions. Also, we have performed whole exome sequencing (WES) in multicase families in order to identify very rare or de novo coding variants with high penetrance.

We have identified a common variant rs4947296 in chromosome 6p21.33, in cases with involvement of both ears, that could define autoimmune inner ear disease. This regulatory variant is an eQTL that influence several genes in the the TWEAK/Fn14 pathway Furthermore, we have defined novel mutations for familial MD in FAM136A, PRKCB, DTNA, DPT and SEMA3D genes in 4 families with autosomal dominant inheritance. Currently, we are assessing the functional role of these genes in a stem cell and zebrafish model.

We are developing an integrated network of molecular pathways, the MDmap, which includes multi-omics data to define endophenotypes according to specific mechanisms of the disease http://disease-maps.org/menieres.

RESEARCH AREAS

Line 1. Genetic contribution to autoimmune Ménière's disease (MD).

The aim of the proposed research is to identify the major genes involved in autoimmune MD as well as the biochemical pathways associated with sporadic MD to decode the driving and modifiers genes of MD.

For this purpose we genotyped by a microarray with 186 loci associated with autoimmune diseases (Inmunochip) in a cohort of patients from Spain and we identified a locus at chromosome 6p21.33 in patients with bilateral sensorineural hearing loss. These locus regulates the expression of HLA class II molecules, as well as several genes in the TWEAK / Fn14 pathway. The carriers of this variant have an NF-kB-mediated inflammatory response in monocytes and lymphocytes.

Line 2. Identification of driver genes in familial Ménière’s disease

Familial MD represents 8-9% of the cases and it has a genetic heterogeneity with autosomal dominant, autosomal recessive and mitochondrial inheritance patterns, being expected an interaction between multiple genes in each family, which would explain the incomplete penetrance or intermediate phenotypes.

Bioinformatic analysis includes the selection of rare variants in our cases to identify candidate genes in each family, and gene-set enrichment analysis to find significant signaling pathways or biochemical common processes.

Our group has sequenced > 100 familial cases and early onset MD to search for an enrichment of rare variants in familial MD.  This work is conducted in a multicenter collaboration with the Baylor-Hopkins Center for Medelian Genomics and the Luxembourg Centre for System Biomedicine. We expect to generate a list of 25-50 candidate genes to define signaling pathways that determine the cochlear and vestibular phenotype of MD. We have identified novel mutations in DTNA, FAM136A, PRCKB, DPT and SEMA3D genes.

In addition, our group has designed two panel of genes related with sensorineural hearing loss and cochlear and supporting cells to find rare variants associated with sporadic MD.

Line 3. Molecular mechanisms of the immune response to molds in MD

The autoimmune inner ear disease (AIED) is characterized by bilateral sensorineural hearing loss with periods of exacerbation caused by a–not-well-known allergic stimulus. This disease is characterized by an immune response in which pro-inflammatory cytokines such as TNFα and IL-1β are released. These cytokines can be released by macrophages or macrophage-like melanocytes of the cochlea that are located in the stria vascularis in the blood-fluid intraestrial barrier. A subset of patients with MD presents an abnormal elevated levels of IL-1β, IL-6 and TNFα. In addition, exposure of mononuclear cells of these patients to fungal extract from could lead to overexpression of IL-1β and IL-6, suggesting an abnormal response to molds.

Our hypothesis is that a significant group of patients with MD have an altered immune response and this subgroup would be part of AIED. The objective is to characterize the cellular subpopulations associated with the proinflamatory response by mass cytometry and single cell RNAseq.

Line 4. Development of a cellular model in familial Meniere's disease.

The main goal of this project is to obtain induced pluripotent stem cells (iPSC) derived from mononuclear cells from individuals with familial MD with the mutations of interest, to prove the persistence of mutations in the iPSC. We have already developed an iPSC model with DTNA and FAM136A mutations. Subsequently, we want to generate otic progenitors to differentiate them into hair cells, auditory and vestibular neurons and epithelial cells with secretory activity.

Line 5. Molecular basis of hiperacusia and tinnitus in Meniere's disease.

Bilateral tinnitus has high heritability. Hyperacusia and tinnitus are constant symptoms in MD. Hypersensitivity and tinnitus poor tolerance reflect an altered brain excitability. The goal of the proposed research is to identify which individuals have a tinnitus problem and to develop a neuronal cell model from iPSC to define the molecular basis of abnormal neuronal excitability.

Our group will select individuals with an extreme phenotype (MD with hyperacusis and unilateral tinnitus persistent without treatment and THI> percentile 90) to undertake a WES study to identify the responsible genes.

Furthermore, our group collaborates with Prof. Christopher Cederroth of Karoliska Institutet in the identification of rare variants in multicase families with tinnitus and high frequency hearing loss.

Line 6: Genetic basis of cerebellar ataxia, neuropathy and bilateral vestibular arreflexia syndrome (CANVAS)

CANVAS disease is a neurodegenerative ganglionopathy with phenotypic heterogeneity. Although most of cases described are sporadic, the findings of affected sibling pairs suggest a familial recessive disorder.

Our group collaborates with Dr. Adolfo Bronstein, from Imperial college in London, in the identification of rare or de novo variants through WES in familiar cases of CANVAS. It has been identified and validated a variant heterozygotes that affect the ELF2 gene. Currently, we are performing functional studies using lentiviral vectors in the BE(2)M17 neuroblastoma cell line.

TECHNOLOGY PLATFORMS

  • Illumina iSCAN: High Performance Genotyping
  • Real-time PCR (Applied Biosystem 7900 y 7500)
  • High throughput DNA sequencing Illumina Nextseq.
  • Bioinformatic analysis data sequencing and prioritization of variants
  • Analysis of gene networks using IPA and MetaCore
  • Cell reprogramming and generation of iPSCs using Sendai virus
  • Directed Mutagenesis and translation of cell lines using lentiviral vectors
  • CyTOF- mass cytometry for single cell proteomics
  • Otoacoustic emissions, auditory and vestibular evoked potentials (Interacoustic)
  • Dynamic posturography (Synapsis)

FUNDING

Title: European School for Interdisciplinary tinnitus research. (Collaborating Researcher)

Principal investigator: Winfried Schlee

Financing entity: EU-H2020                                                        Code: MSCA-ITN-722046

Period: 2017-2020

Funding: 3.823.897€

 

Title: Validación de un marcador genético regulador de la inflamación para el diagnóstico de enfermedad de Meniere autoinmune (DIEMA).

Principal investigator: Jose Antonio Lopez Escamez

Financing entity: Programa INNBIO-ADVANCE, FIBAO.     Code: INB-007

Period: 2017-2019

Funding:  14.000€

 

Title: Designing a molecular for Meniere’s disease (MD-CONNECT)

Principal investigator: Reinhard Schneider/Jose Antonio Lopez Escamez

Financing entity: FNR                                                                  Code: INTER/Mobility/17/11772209

Period: 2017-2018

Funding:  160.060€

 

Title: Papel de la variante reguladora rs4947296 y la respuesta proinflamatoria en la enfermedad de Meniere.

Principal investigator: Jose Antonio Lopez Escamez

Financing entity: Instituto de Salud Carlos III                       Code: PI17/01644                             

Period: 2018-2020

Funding:  171.820€

 

Title: Desarrollo de Un Modelo de Enfermedad de Ménière Familiar en Pez Cebra Mediante Edición Génica Dirigida Con Crispr/Cas9.

Principal investigator: Maria Teresa Requena Navarro

Financing entity: Consejeria de Salud                                  Code: EF-0023-2017

Period: 2018

Funding:  15.026€

SELECTED PUBLICATIONS

  • Requena T, Cabrera S, Martin-Sierra C, Price SD, Lysakowski A, Lopez-Escamez JA. Identification of two novel mutations in FAM136A and DTNA genes in autosomal dominant Meniere’s disease. Hum Mol Genet 2014 2015 Feb 14; 24(4):1119-26. http://www.ncbi.nlm.nih.gov/pubmed/25305078 IF (JCR) – Quartile (JCR) 6.393- Q1
  • Martin-Sierra C, Requena T, Frejo L, Price SD, Gallego-Martinez A, Batuecas-Caletrio A, Santos-Perez S, Soto-Varela A, Lysakowski A, Lopez-Escamez JA. A novel missense varint in PRKCB segregates low-frequency hearing loss in an autosomal dominant family with Meniere’s disease. Hum Mol Genet 2016  IF(JCR)- Quartile (JCR) 6.393-Q1-D1
  • Martin-Sierra C, Gallego-Martinez A, Requena T, Frejo L, Batuecas-Caletrío A, Lopez-Escamez JA. Variable expressivity and genetic heterogeneity involving DPT and SEMA3D genes in autosomal dominant familial Meniere’s disease. Eur J Hum Genet 2016  IF(JCR)- Quartile (JCR) 4.580-Q1
  • Frejo L, Soto-Varela A, Santos-Perez S, Aran I, Batuecas-Caletrio A, Perez-Guillen V, Perez-Garrigues H, Fraile J, Martin-Sanz E, Tapia MC, Trinidad G, Garcia-Arumi AM, Gonzalez-Aguado R, Espinosa-Sanchez JM, Marques P, Perez P, Benitez J, Lopez-Escamez JA. Clinical subgroups in bilateral Meniere’s disease. Front Neurol 2016, 24 Oct doi: 10.3389/fneur.2016.00182. IF(JCR)- Quartile (JCR) 3.184-Q2
  • Maas IL, Brüggemann P, Requena T, Bulla J, Edvall NK, Hjelmborg JvB, Szczepek AJ, Canlon B, Mazurek B, Lopez-Escamez JA, Cederroth CR. Genetic susceptibility to bilateral tinnitus in a Swedish twin cohort. Genet Med 2017, Mar23 doi: 10.1038/gim.2017.4 IF(JCR)- Quartile (JCR) 8.229-D1
  • Requena T, Gallego-Martinez A, Lopez-Escamez JA. A pipeline combining multiple strategies for prioritizing heterozygous variants for the identification of candidate genes in exome datasets. Hum Genomics 2017 11:11. DOI: 10.1186/s40246-017-0107-5. IF(JCR)- Quartile (JCR) 3.327-Q2
  • Roman-Naranjo P, Gallego-Martinez A, Lopez-Escamez JA. Genetics of vestibular syndromes. Curr Opin Neurol. 2018 Feb;31(1):105–10. IF(JCR)- Quartile (JCR) 4.699-Q1
  • Frejo L, Requena T, Okawa S, Gallego-Martin A, Martinez-Bueno M, Aran I, Batuecas-Caletrio A, Benitez J, Espinosa-Sanchez JM, Fraile-Rodrigo J, Garcia-Arumi AM, Gonzalez-Aguado R, Marques P, Martin-Sanz E, Perez-Fernandez N, Perez P, Perez-Garrigues H, Santos-Perez S, Soto-Varela A, Tapia MC, Trinidad G, del Sol A, Alarcon-Riquelme ME, Lopez-Escamez JA. Regulation of Fn14 receptor and NFkB underlies inflammation in Meniere disease. Front Immunol 2017 https://doi.org/10.3389/fimmu.2017.01739 IF(JCR)- Quartile (JCR) 6.429-Q1
  • Schlee W, Hall DA, Canlon B, Cima RF, De Kleine E, Hauck F, Huber A, Gallus S, Kleinjung T, Kypraos T, Langguth B, Lopez.Escamez JA, Lugo A, Meyer M, Mielczark M, Norena A, Pfiffner F,  Pryss RC, Reinchert M, Requena T, Schecklmann M, Van Dijk P, Van de Heyning P, Weisz N, Cederroth CR. Innovations in Doctoral Training and Research on Tinnitus: The European School on Interdisciplinary Tinnitus Research (ESIT) Perspective. Front Neurosc 2017. doi: 10.3389/fnagi.2017.00447. IF(JCR)- Quartile (JCR) 4.504-Q1
  • Frejo L, Gallego-Martinez A, Requena T, Martin-Sanz E, Amor-Dorado JC, Soto-Varela A, Santos-Perez S, Espinosa-Sanchez JM, Batuecas-Caletrio A, Aran I, Fraile J, Rossi-Izquierdo M, Lopez-Escamez JA. Proinflammatory cytokines and response to molds in mononuclear cells of patients with Meniere disease. Scientific Rep 2018 8, Art 5974. doi:10.1038/s41598-018-23911-4. IF (JCR)-Quartile (JCR) 4,280-Q1

RESEARCH GROUP PATENTS

Application Number:

P2011132013

Priority date:

14.12.2011

Title:

Method to obtain useful data for diagnosis and prognosis of sensorineural hearing loss

Authors:

Lopez-Escamez JA, Lopez-Nevot MA, Gazquez I, Moreno A, Aran I, Soto-Varela A, Santos S, Perez-Garrigues H, Lopez-Nevot A, Requena T.

Applicant:

Fundación Progreso y Salud /Agencia Publica Hospital de Poniente

 

Application Number:

P201430716

Priority date:

16.05.2014

Títle:

Using polymorphism (SNPs) of the NFKB1 gene for auditory prognosis in Meniere disease.

Authors:

Lopez-Escamez JA, Cabrera S, Sanchez E, , Alarcon-Riquelme ME, Espinosa-Sanchez JM, Requena T.

 

 

Applicant:

Fundación Progreso y Salud /Servicio Andaluz de Salud

Application Number:

P201531458

Priority date:

09.10.2015

Títle:

Using allelic variants in the region 6p21.33 for the diagnosis and prognosis of Meniere’s disease

Authors:

Lopez-Escamez JA, Cabrera S, Alarcon-Riquelme ME, Requena T.

Applicant:

Fundación Progreso y Salud /Servicio Andaluz de Salud

 

 

Application Number:

P201630745

Priority date:

03.06.2016

Title:

Proinflammatory cytokines as diagnostic marker in episodic vestibular syndrome

Authors:

Lopez-Escamez JA, Frejo Navarro L, Espinosa-Sanchez Juan Manuel, Requena T.

Applicant:

Servicio Andaluz de Salud

ADDITIONAL INFORMATION

Doctoral theses supervised

  • Monitoring of vestibular function and quality of life related to health in patients with paroxysmal benign positional vertigo. Maria Isabel Molina Palma.14/06/2007.
  • Immunogenetic basis of Meniere's disease: searching for genetic markers. Irene Gazquez Perez. 24/09/2012.
  • Identification and validation of mutations in familial Ménière's disease. Teresa Requena Navarro. 02/10/2015.
  • Mechanisms of inflammations in Meniere’s disease. Lidia Frejo Navarro. 12/05/2017.

Scientific Societies:

  • Program Committee for Association for Research in Otolaryngology. 2016-2019.
  • Spanish Society of Otorhinolaryngology and Cervical-Facial Pathology. 1995-2014.
  • International Committee for the Classification of vestibular disorders of the Barany Society. 2008-2017.
  • Royal Academy of Medicine of Eastern Andalucía. 2013-2017
  • European Society of Human Genetics. 2017.

Research Networks:

  • Better Understanding the Heterogeneity of Tinnitus to Improve and Develop New Treatments (TINNET). Chair: Berthold Langguth. COST Action BM1306. 2014-2017
  • European School for Interdisciplinary Tinnitus Research (ESIT). Chair: Winfried Schlee. H2020-MSCA-ITN 722046.. 2017-2020.
  • Meniere’s disease Consortium (MediC): European Network for the study of the Meniere’s disease. 2016.

 

Continued scientific collaborations with:

  • University of Illinois, Chicago, USA
  • Luxembourg Centre for System Biomedicine- Centre Hospitalier Emile Mayrisch, Luxembourg
  • San Raffaele Hospital, Milano, Italy
  • Imperial College London, UK
  • Karolinska Institutet, Stockholm, Sweden
  • Baylor-Hopkins Center for Mendelian Genomics, Baltimore, USA.
  • Ludwig-Maximillian University, München, Germany

EDITORIAL BOARD MEMBERSHIP

  1. Acta Otorrinolaringológica Española. 2013
  2. Audiology Research. 2014
  3. Current Otorhinolaryngology Reports. 2013-2018
  4. Frontiers in Neurology-Neurotology Section. 2016-2018
  5. Scientific Reports. 2015-2018
  6. International Journal of Otolaryngology. 2015

Reviewer of the following Journals

  • The Laryngoscope. Lippincott Williams & Wilkins. 2004-11.
  • Brain. A Journal of Neurology. 2007-2009
  • Journal Neurology Neurosurgery & Psychiatry. BMJ Group. 2007
  • Acta Otorrinolaringologica Española. 2007-2018
  • Molecular Neurobiology. 2009.
  • Molecular Biology Reports. 2010.
  • International Journal Otolaryngology. 2011
  • Current Genomics. 2011
  • PloS ONE. 2011-2018
  • Pharmacogenetics and genomics. 2012.
  • Ear and Hearing. 2012-2013
  • International Journal of Audiology. 2012
  • European Journal of Medical Genetics. 2012-13.
  • Journal of Clinical Immunology. 2013.
  • Biochimica and Biophysica Acta- Molecular Basis of Disease. 2013.
  • British Medical Journal. 2016
  • Hearing Research. 2015-17
  • Scientific Reports. 2015-18