Development of Partner Diagnostics and Therapeutics for Genetic Eye Disease

  • Connie Chao-Shern

Student thesis: Doctoral Thesis


Approximately 165 of the 650 primary ocular diseases are contributed to corneal abnormalities (1). They include diseases such as the TGFBI corneal dystrophies, Keratoconus (KC), Fuchs’ Endothelial Corneal Dystrophy (FECD), Meesmann Epithelial corneal dystrophy (MECD) and Brittle cornea syndrome (BSC) (1). Mutations in the transforming growth factor beta induced (TGFBI) gene cause a spectrum of distinct epithelial and stromal corneal dystrophies with corneal amyloid and non-amyloid deposits (2). KC is a much more complex disease and not only is it less understood, it is most likely a disease influenced by complex genetic and environmental factors. FECD is the most commonly inherited corneal disease; however, the degrees of disease manifestation are impacted by genetic disposition and environmental factors, even within members of the same family (3).
This thesis describes the progression of partner diagnostic testing methods for inherited corneal diseases evolving from real-time PCR to Next Generation Sequencing (NGS). Literature reviews were performed on genetic corneal diseases leading to a six mutation Single Nucleotide Polymorphism (SNP) panel real-time PCR test that addresses a gap in the diagnostic capability. KC was studied with a cohort of diseased and normal individuals and a targeted NGS panel test with 75 genes was developed. It offers the detection of 70+ mutations within the TGFBI gene, and 1,000+ variants across the 75 genes. Building upon the knowledge of the partner diagnostic testing, a personalized gene therapy and gene editing treatment plan can be created accordingly. This thesis also reviews the laboratory processes for gene editing of the target mutations. There are limitations in the 75-gene panel for KC. More work needs to be done to discover novel variants. To overcome the limitation of editing one mutation at a time, a more effective gene editing method should be explored to treat multiple mutations simultaneously.
Date of AwardAug 2019
Original languageEnglish
SupervisorAndrew Nesbit (Supervisor), Paul Thompson (Supervisor) & Tara Moore (Supervisor)


  • TGFBI gene
  • TGFBI Corneal Dystrophy
  • Keratoconus
  • Genetic testing
  • PCR
  • Next Generation Sequencing

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