Controlling donor and newborn neuron migration and maturation in the eye through microenvironment engineering

Area of Science:

• Neuroscience
• Regenerative Medicine
• Ophthalmology

Background:

• Cell therapy requires precise control of donor cell behavior in recipient tissues.
• The eye offers a unique model for studying central nervous system neuron fate and function.
• Retinal ganglion cells (RGCs) are lost in optic neuropathies, necessitating replacement strategies.

Purpose of the Study:

• To develop a methodology for guiding stem cell-derived and endogenously regenerated neurons.
• To overcome barriers in RGC integration, specifically cell migration, for successful retinal circuitry repair.
• To identify and apply soluble ligands for controlling donor cell function post-transplantation.

Main Methods:

• In silico analysis of the developing human retina's single-cell transcriptome to identify receptor-ligand candidates.
• In vitro functional assays to test the ability of identified candidates to guide human stem cell-derived RGCs.
• Engineering an artificial gradient using the lead molecule SDF1 in the retina for in vivo studies.

Main Results:

• Six receptor-ligand candidates were identified, with SDF1 selected as the lead molecule.
• An artificial SDF1 gradient increased donor RGC migration into the ganglion cell layer (GCL) by 2.7-fold.
• Increased displacement of newborn RGCs from the inner nuclear layer (3.3-fold) and expression of mature RGC markers in GCL-integrated cells were observed.

Conclusions:

• A novel 'in silico-in vitro-in vivo' framework was established for identifying and applying soluble ligands.
• Engineering the microenvironment with SDF1 effectively guides RGC migration and promotes integration.
• This approach holds promise for improving cell transplantation outcomes in treating optic neuropathies.