Research team
Expertise
Breeding and handling of transgenic animal models for hereditary hearing loss and vestibular dysfunction. Performing auditory brainstem response and distortion oto-acoustic emission measurements to assess hearing function in rodents. Investigating balance through measurement of the vestibular ocular reflex and observation of reflexes and behavior. Noise exposure in mice. Injection of molecules into the inner ear using the posterior semicircular canal approach in adult mice. Development of gene therapy to tackle hearing and vestibular loss in DFNA9 patients using a new humanized DFNA9 mouse model. Experience in immunohistochemistry (cryosections and whole mount) of the Organ of Corti and the spiral ligament to visualize different cell types, inner ear proteins and inner ear inflammation using specific antibodies. Molecular biology techniques involving RNA/DNA isolation, PCR, sequencing, ....
Fine-tuning delivery methods for the use of antisense oligonucleotides to modify progression of sensorineural hearing loss.
Abstract
Hearing loss is the most common sensory deficit and has been recognized by the WHO as a priority disease for research into therapeutic interventions. Deafness Autosomal Dominant 9 (DFNA9) is one of the best-studied forms of dominantly-inherited, adult-onset hearing loss and is characterized by progressive sensorineural hearing loss (SNHL). It is caused by mutations in the COCH gene, which encodes for the protein cochlin. In this project, the applicant presents the development and pre-clinical testing of an optimized delivery method for RNA therapeutics for hearing loss. The applicant will use DFNA9 and the antisense oligonucleotides (ASOs) she validated in vitro, as a case study for this novel treatment paradigm. More specifically, the applicant will go beyond the current state of the art by benchmarking the clinically relevant/feasible intratympanic delivery method and generating a purpose-built hydrogel for ASO treatment. Furthermore, cell-specific uptake of different gapmer ASO chemistries in the inner ear will be studied to achieve targeted delivery. Finally, the applicant will generate data on the pharmacokinetics, safety and efficacy of (gapmer) ASOs in the mammalian inner ear of our genetically engineered humanized DFNA9 mouse model. As such, this project will pave the way for clinical applications of ASOs and other types of gene therapy to target the cochlea for the future treatment of DFNA9 and other inner ear disorders.Researcher(s)
- Promoter: Van Rompaey Vincent
- Co-promoter: Verdoodt Dorien
- Fellow: Aben Fien
Research team(s)
Project type(s)
- Research Project
Delivery of CRISPR-Cas9 mRNA-based therapy into the inner ear by lipid nanoparticles: Cure deafness by disrupting mutant COCH expression in a humanized DFNA9 mouse model.
Abstract
Hearing loss has a significant impact on quality of life and cognitive function, affecting more than 1.57 billion people worldwide. The application will focus on the autosomal dominant disorder DFNA9 which leads to post-lingual severe-to-profound sensorineural hearing loss (SNHL). DFNA9 is caused by heterozygous mutations in the COCH gene that lead to the formation of mutant cochlin. Currently, no treatment is available to prevent or cure SNHL in DFNA9. This application presents an innovative approach to go beyond the current state-of-art in the field of SNHL by in vivo evaluation of a disease-modifying gene therapy injected in the inner ear to downregulate expression of the mutant COCH allele. The lab will do so by using lipid nanoparticles loaded with a CRISPR-nuclease optimized in vitro in a unique genetically humanized DFNA9 mouse model carrying the exact mutation of our human population. By sparing the normal allele, a heterozygous protein-truncating mutation will be created, which would not lead to a phenotype of SNHL.Researcher(s)
- Promoter: Van Rompaey Vincent
- Co-promoter: Verdoodt Dorien
- Fellow: Sels Lize
Research team(s)
Project type(s)
- Research Project
Development of a CRISPR-nuclease gene therapy to cure deafness in DFNA9 patients.
Abstract
Hearing impairment is the most frequent sensory deficit in the human population, affecting 440 million people worldwide, whereby loss of hearing and balance has a significant impact on quality of life and society. Hearing loss is also listed by the World Health Organization as a priority disease for research into therapeutic interventions to address public health needs. DFNA9 (DeaFNess Autosomal 9) is an autosomal dominant hearing disorder caused by a heterozygous gain-of-function mutation in the COCH gene (Coagulation Factor C Homology) and is characterized by progressive late-onset (3rd-5th decade) sensorineural hearing loss (SNHL) and deafness. Within Belgium and the Netherlands, there are >1000 patients affected by the p.P51S COCH mutation, who – in the current absence of a disease modifying therapy – will develop deafness. A promising approach to tackle the pathophysiological mechanism of DFNA9 and stop or delay the onset of SNHL is to introduce a CRISPR-Cas9-based therapy into the inner ear, delivered by an AAV vector to disrupt or downregulate the expression of (mutant) COCH. In this project, I will phenotype a newly established humanized DFNA9 mouse model and assess whether in vivo delivery of a CRISPR-nuclease using an AAV-based viral vector leads to specific disruption of COCH.Researcher(s)
- Promoter: Van Rompaey Vincent
- Fellow: Verdoodt Dorien
Research team(s)
Project type(s)
- Research Project
Towards a cure for deafness in DFNA9: loading lipid nanoparticles with CRISPR-Cas9 to safely target dominant-negative mutations in the COCH gene in vitro and in vivo.
Abstract
Hearing impairment is the most frequent sensory deficit in the human population, affecting 440 million people worldwide, whereby loss of hearing and balance has a significant impact on quality of life and society. Hearing loss is also listed by the World Health Organization as a priority disease for research into therapeutic interventions to address public health needs. DFNA9 (DeaFNess Autosomal 9) is an autosomal dominant hearing disorder caused by different heterozygous gain-of-function mutations in the COCH gene (Coagulation Factor C Homology) and is characterized by vestibular dysfunction and progressive late-onset (3rd-5th decade) sensorineural hearing loss (SNHL) leading to deafness. Within Belgium and the Netherlands, there are >1000 patients affected by the p.P51S COCH mutation, one of the heterozygous mutations causing DFNA9, who – in the current absence of a disease modifying therapy – will develop deafness. A promising approach to tackle the pathophysiological mechanism of DFNA9 and stop or delay the onset of SNHL is to introduce a CRISPR-Cas9 mRNA based therapy into the inner ear, delivered by a Lipid nanoparticle (LNP) to disrupt or downregulate the expression of mutant COCH protein. Until today, most researchers focus on the use of viral vectors, mostly AAV, to apply gene therapy in vivo in the inner ear. However, many people have already been exposed to AAV earlier in life and generated specific antibodies. Therefore, using LNP could provide a suitable alternative as they are already been proven to be safe when injecting a large population (COVID-19 mRNA vaccines). Moreover, using Cas9 mRNA instead of Cas9 DNA will only lead to a temporary expression of the Cas9 protein which will largely decrease the change on an evoked immune response against Cas9. To develop a cochlear gene therapy than can be translated to a human setting, a highly innovative humanized DFNA9 mouse model was generated. These mice bear humanized COCH harbouring the p.P51S mutation and thus mimic the genotype of DFNA9 patients. The goal of this project is: (1) assessing the safety and targeted delivery of LNP injected in vivo in the inner ear of our humanized DFNA9 mouse model (2) assess the use of LNP for in vitro delivery of Cas9/gRNA complexes to specifically downregulate the production of mutant cochlin in embryonic fibroblasts derived from our humanized DFNA9 mouse model.Researcher(s)
- Promoter: Verdoodt Dorien
Research team(s)
Project type(s)
- Research Project
Inner ear gene therapy to prevent deafness in DFNA9.
Abstract
Hearing loss has a significant impact on quality of life and society in general. Hearing impairment is the most frequent sensory deficit in human populations, affecting 360 million people worldwide. It was listed by the World Health Organization as one of the priority diseases for research into therapeutic interventions to address public health needs. DFNA9 is a dominant hereditary disorder, caused by heterozygote mutations in the COCH gene, which progressively leads to bilateral deafness and balance loss by the age of 50-70 years. Currently, no treatment is available to prevent hearing loss or balance loss in DFNA9 patients. Local gene therapy to restore hearing or prevent hearing loss has been studied in neonatal mouse models for several years. Currently, a clinical study is ongoing in adult patients with profound hearing loss to restore hair cells by injecting virus-based vectors -carrying correcting genetic information- directly into the inner ear. In this project, we aim to generate an inner ear gene therapy tool to prevent hearing loss in a pre-clinical mouse model of DNFA9. Using Adeno-associated virus (AAV)-based vectors, we will apply CRISPR-Cpf1 genome engineering technology to target directly within in the cochlea Coch genomic DNA in a safe and effective way in order to disrupt expression of the mutant (and wild type) Coch protein before onset of disease. Hereby, we expect to reduce or prevent DNFA9-associated sensorineural hearing loss.Researcher(s)
- Promoter: Van Rompaey Vincent
- Co-promoter: Ponsaerts Peter
- Fellow: Verdoodt Dorien
Research team(s)
Project type(s)
- Research Project