PROJECT 2

Functional Assessment of Glaucoma Candidate Disease Genes using Genetically Modified Retinal Organoids

POSITION FILLED

Why?
Previous genomics research showed that glaucoma is caused by at least 140 candidate disease genes. While this knowledge facilitates generic human risk assessments and has yielded extra-ordinary new insight in the etiology of the disease, little is known about the function of these genes, how they potentially interact and to what extent they contribute to glaucoma. Up to 10 of these 140 candidate genes consistently show up in all genomic studies and are expressed in the retina, and will be the focus of study in this WP. The objective of this WP is to study the function, molecular program, and interaction of (up to) ten of these glaucoma disease genes, using an already established CRISPR/Cas9 human retinal organoid pipeline. This project will yield fundamental new insights in the etiology of glaucoma, and may be the basis for novel rational -omics driven therapeutic designs.

How?
In this project, the PhD will generate and characterize (new) human retinal organoid based glaucoma models and translate findings into rational therapeutic designs. Initially, an already in-house established retinal organoid glaucoma model (optic nerve-in-a-dish model) will be characterized. This stem cell derived model is based on CRISPR/Cas9 deletion of a gene involved in a mendelian form of early onset glaucoma. Its characterization is expected to elucidate (part of) the molecular machinery, including the role of the aforementioned “selected up to ten” genes, and cellular pathology underlying retinal damage in glaucoma. Cutting-edge biological read-outs are established in the lab and AUMC facilities and include advanced microscopy, DNA and RNAseq, protein analysis, immunohistochemistry, ganglion cell subtype analysis, ganglion cell outgrowth assays, cellular imaging, bioinformatics and electrophysiology. In the second part of the project new organoid models will be made, (medicine) tested and characterized.

What can you expect?
The successful candidate will be gain expertise in molecular and cellular neuro developmental biology and neurodegeneration of the retina and optic nerve. The candidate will be trained in advanced molecular and cellular approaches based on human in vitro modelling of human retinal and optic nerve (glaucomatous) organoids. In addition, the candidate will benefit from the training activities scheduled by the EGRET-AAA program (workshops, meetings, etc.). After successful completion of the PhD, the trainee will obtain a joint doctorate of Amsterdam University and the Sorbonne University, Paris.

Where?
This project is a joint-doctorate between the Amsterdam UMC and the Vision Institute (Sorbonne University, Paris). At AUMC, the trainee will be daily supervised by Dr. Ellie Wagstaff at the Bergen Ophthalmogenetics lab (https://ophthalmogenetics.nl) AmsterdamUMC). For direct clinical and pathological glaucoma issues, Dr. Sarah Janssen (Ophthalmology department AUMC) is involved. The AUMC offers all knowledge and facilities needed for this project. The trainee can participate in the world-leading Neuroscience School Amsterdam, and will conduct a short secondment project in the Nicol lab (Paris, Sorbonne UNI), to learn ins and outs of neural outgrowth of retinal explants.

Who are we looking for?
We are looking for a highly motivated, dynamic and talented student who enjoys teamwork and has strong communication and interpersonal skills. The successful candidate must preferentially hold a laboratory based Neuroscience, Vision research or Molecular cell Biology Master obtained in the 2022-2023 academic year. Fluency in English is mandatory.

References

  • The vast complexity of primary open angle glaucoma: disease genes, risks, molecular mechanisms and pathobiology. Janssen SF, Gorgels TG, Ramdas WD, Klaver CC, van Duijn CM, Jansonius NM, Bergen AA. Prog Retin Eye Res. 2013 Nov;37:31-67.
    PMID: 24055863. https://pubmed.ncbi.nlm.nih.gov/24055863/
  • An alternative approach to produce versatile retinal organoids with accelerated ganglion cell development. Wagstaff PE, Ten Asbroek ALMA, Ten Brink JB, Jansonius NM, Bergen AAB.
    Sci Rep. 2021 Jan 13;11(1):1101. PMID: 33441707. https://pubmed.ncbi.nlm.nih.gov/33441707/
  • The Role of Small Molecules and Their Effect on the Molecular Mechanisms of Early Retinal Organoid Development. Wagstaff PE, Heredero Berzal A, Boon CJF, Quinn PMJ, Ten Asbroek ALMA, Bergen AA. Int J Mol Sci. 2021 Jun 30;22(13):7081. PMID: 34209272.\
    https://pubmed.ncbi.nlm.nih.gov/34209272/

 POSITION FILLED