Discovering differentially expressed genes in feline tooth resorption (TR) using RNA sequencing (RNA seq)

Project Details

Description

Hypothesis: We will test the hypothesis that cats affected with feline TR have genetic aberrations causing dysregulation of normal odontoclastic function.
Aim of pilot study: (1) Collect five TR affected and five TR negative tooth samples from cats (2) Extract RNA with RNA Integrity Number (RIN) values above 7 (3) Use RNAseq of teeth from TR affected and unaffected cats to identify differentially expressed genes in the local tooth microenvironment.
Preliminary data: We have so far collected twelve tooth samples (nine TR affected and three TR free animals). It was challenging to extract RNA of adequate quality suitable for RNAseq (RIN >7), but we have now established a tooth sampling procedure and RNA extraction protocol that allows for this

Layman's description

Under normal circumstances, kittens shed their milk teeth which are then replaced by permanent adult teeth. In order for the developing adult tooth to erupt from its bony pocket in which it develops the overlying milk tooth and bone have to be resorbed. Specialized cells called odontoclasts are crucial for tooth eruption as they eat mineralised tissues of bone and teeth. When all the milk teeth are shed, these cells should stop resorbing but in up to 30 % of cats these cells are reactivated and they start to attack the permanent dentition. This is feline tooth resorption (TR), a common cause of tooth loss and mouth pain in cats. The condition is progressive and the only treatment is to extract the affected tooth. Severely affected animals can lose almost all their teeth.
A similar condition in man has got a genetic predisposition and epidemiological studies in cats suggest a higher incidence in purebred cats and show a widespread presence in siblings, indicating genetics may play a role in the disease development in cats too.
This pilot study will use a new powerful technique called RNA sequencing (RNAseq). It allows us to get a snapshot of the genes that are active in a given sample. We will sequence teeth from TR affected and disease free animals, and measure which genes are more active in teeth affected with TR. These genes can then be further investigated to look for causative mutations in a much larger DNA sequencing study.
AcronymFiona & Ian Russel Seed Corn Fund for Companion Animal Research
StatusFinished
Effective start/end date1/06/1515/01/17