grupo

Group name: Otology & Neurotology- Genomics of vestibular disorders

    Group leader
  • Jose Antonio López Escámez
  • Senior Researcher
  • Miguel Ángel López Nevot
  • Postdoctorals
  • Maria Teresa Requena Navarro
  • PHD Students
  • Álvaro Gallego Martínez
  • Juan Manuel Espinosa Sánchez
  • Lidia Frejo Navarro
  • Internships
  • Ivan Morales Esquina
  • Pablo Roman-Naranjo Varela

Presentation Video

SCIENTIFIC INTEREST AREAS

Recurrent vertigo is a common health problem caused by several disorders of the vestibular system in the inner ear. 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.

MD is a multifactorial chronic disorder in 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 autosomal dominant, autosomal recessive and mitochondrial inheritance patterns. Our group has compiled the largest collection of human DNA samples from MD patients including >100 multicase families and > 2000 sporadic cases to identify the genes associated with Meniere’s disease. 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 variants with high penetrancy in coding regions.

We have identified several loci in chromosome 6p21.33, in cases with involvement of both ears, that could define autoimmune inner ear disease. Furthermore, we have defined novel mutations for familial MD in FAM136A, PRKCB, DTNA, DPT and SEMA3D genes in three families with autosomal dominant inheritance. Currently, we are assessing the functional role of these genes in a stem cell model with the variants of interest.

RESEARCH AREAS

Line 1. Genomic architecture of sporadic Ménière's disease (MD).

The aim of the proposed research is to identify the major genes involved in 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. This project is made in collaboration with Prof. Marta Alarcón group.

Also, our group analyses the gene interaction between the variants discovered using bioinformatics analysis of gene networks and functional analysis in cellular models with the candidate variants.

Line 2. Identification of novel mutations in familial Ménière’s disease

Familial Menière’s disease 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 exclusive variants in our cases to identify candidate genes in each family, and biochemical-based pathway analyses to define groups of coding genes with candidate variants prioritized located in signaling pathways or biochemical common processes.

Our group is sequencing > 15 multicase families and 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 a panel of genes related with sensorineural hearing loss to find rare variants associated with sporadic MD through DNA-seq.

Line 3. Molecular mechanisms in the autoimmune inner ear disease (AIED)

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. Thus, AIED would develop by the existence of epitopes in environmental antigens with cross-reactivity with the inner ear proteins. The patients with AIED 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.

Menière’s disease as well as AIED can react to corticoids treatment. Therefore, several studies have described partial audiometric improvements in 58-72% of cases with bilateral hearing loss and in 30% of cases with unilateral hearing loss. This response to steroids depends on the expression of the soluble form of type 2 IL-1 receptor (sIL1R2), which is a decoy receptor that binds to IL-1β irreversibly and inhibits the signal of IL-1β.

Our hypothesis is that a significant group of patients with MD have a genetic susceptibility determining an altered immune response and this subgroup would be part of AIED. The objective is to develop a diagnostic and prognostic tool to identify patients with AIED, and their possible treatment with antibodies against IL-1β receptor.

Line 4. Development of a cellular model in 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. Subsequently, we want to generate otic progenitors to differentiate them into hair cells, auditory and vestibular neurons and epithelial cells with secretory activity. This project is carried out in collaboration with Dr. Pedro Real’s group.

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 sequencing SOLID4 5500xl, 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
  • Video-oculography 2D and 3D with head accelerometer (SMI)
  • Otoacoustic emissions, auditory and vestibular evoked potentials (Bio-Logic)
  • Dynamic posturography (Synapsis)

FUNDING

Title: Better Understanding the Heterogeneity of Tinnitus to Improve and Develop New Treatments (TINNET) (Collaborating Researcher)

Principal investigator: Winfried Schlee

Financing entity: COST Action.                                                   Code: BM1306

Period: 2014-2017

Funding: 510.000 €

 

Title: Validación de los genes FAM136A, PRKCB, THAP1 y GRHL2 mediante secuenciación dirigida en la enfermedad de Meniere

Principal investigator: Teresa Requena

Financing entity: FPS                                                    Code: PI-0496-2015

Period: 2015-2017

Funding:  34.000 €

 

Title: Human iPSC-based model to develop a personalized treatment for Meniere’s Disease (MD) by sustained target gene-knockdown

Principal investigator: AeRi Ji

Financing entity: BD Bioscience Research Program 2015                                  

Period: 2015-2016

Funding:  10.000 $

 

Title: Assessment of familial Meniere’s disease genes by target sequencing and induced pluripotent stem cells (iPSC)-based cellular models (Meniere’s cell).

Principal investigator : Jose Antonio Lopez Escamez

Financing entity: Ménière’s Society Grant, UK  Code: PI-Meniere’s cell/Ménière’s Society

Period: 2016-2017

Funding:  39.687 GBP

 

Title: Better Understanding the Heterogeneity of Tinnitus to Improve and Develop New Treatments (TINNET). (Investigador colaborador).

Principal investigator: Winfried Schlee

Financing entity: European School for Interdisciplinary Tinnitus Research (ESIT)

Code: H2020-MSCA-ITN 722046

Period: 2017-2020

Funding:  3,823,897€

SELECTED PUBLICATIONS

  1. Requena T, Espinosa-Sanchez JM, Lopez-Escamez JA. Genetics of dizziness: cerebellar and vestibular disorders. Current Opinion Neurology 2014; 27(1): 98-104. http://www.ncbi.nlm.nih.gov/pubmed/24275721 IF (JCR) – Quartile (JCR)  5,416 – Q1
  2. 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. Human Molecular Genetics 2014 2015 Feb 14; 24(4):1119-26. http://www.ncbi.nlm.nih.gov/pubmed/25305078 IF (JCR) – Quartile (JCR) 6.393- Q1
  3. Lopez-Escamez JA, Carey J, Chung WH, Goebel JA, Magnusson M, Mandala M, Newman-Toker DE, Strupp M, Suzuki M, Trabalzini F, Bisdorff A. Diagnostic criteria for Meniere’s disease. J Vest Res 2015; 25 (1): 1-7. IF (JCR)-Quartile (JCR) 1.190-Q2.
  4. Cabrera S, Ji AE, Frejo L, Ramos-Mejia V, Real P, Lopez-Escamez JA. Generation of human iPSC line GRX-MCiPS4F-A2 from adult peripheral blood mononuclear cells (PBMCs) with Spanish genetic background. Stem Cell Report 2015 IF (JCR)-Quartile (JCR) 3.693-Q1
  5. Frejo L, Giegling I, Teggi R, Lopez-Escamez JA, Rujescu D. Genetics of vestibular disorders: pathophysiological insights. J Neurol. 2016 Apr;263(Suppl 1):45–53.  IF (JCR)-Quartile (JCR) 3.377-Q2.
  6. 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
  7. 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
  8. Martin-Sanz E, Vaduva C, Esteban J, Sanz R, Lopez-Escamez JA. High frequency sensorineural hearing loss associated with vestibular episodic síndrome. Clin Otolaryngol 2016 DOI: 10.1111/coa.12710 IF(JCR)- Quartile (JCR) 2.627-Q1
  9. Lopez-Escamez JA, Bibas T, Cima RFF, Van de Heyning P, Knipper M, Mazurek B, Szczepek AJ, Cederroth CR. Genetics of tinnitus: an emerging area for molecular diagnosis and drug development. Front Neurosci 2016 .IF(JCR)- Quartile (JCR) 3.398-Q2
  10.  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

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.

Scientific Societies:

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

Research Networks:

  • Better Understanding the Heterogeneity of Tinnitus to Improve and Develop New Treatments (TINNET). Chair: Berthold Langguth. COST Action BM1306. 2014-2017
  • Better Understanding the Heterogeneity of Tinnitus to Improve and Develop New Treatments (TINNET). Chair: Winfried Schlee. H2020-MSCA-ITN 722046. European School for Interdisciplinary Tinnitus Research (ESIT). 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.

EDITORIAL BOARD MEMBERSHIP

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

Reviewer of the following Journals

  • The Laryngoscope. Lippincott Williams & Wilkins. 2004-11.
  • Canadian Medical Association Journal. 2006-11
  • Brain. A Journal of Neurology. 2007-2009
  • Journal Neurology Neurosurgery & Psychiatry. BMJ Group. 2007
  • Acta Otorrinolaringologica Española. 2007-11
  • Molecular Neurobiology. 2009.
  • Molecular Biology Reports. 2010.
  • Journal of Postgraduate Medicine. 2010.
  • Future Neurology. 2010.
  • International Journal Otolaryngology. 2011
  • Current Genomics. 2011
  • PloS ONE. 2011-2014
  • 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.
  • Hearing Research. 2015-16
  • Scientific Reports. 2015-16