Year 1

There is currently no effective treatment to restore or improve vision for patients suffering from blinding diseases such as dry, age-related macular degeneration and retinitis pigmentosa. Alleviation of these diseases requires both new photoreceptors and retinal pigment epithelium (RPE). However, a unique method to transplant fetal retinal progenitor sheets together with its supporting retinal pigment epithelium has been shown to improve vision in animal models of retinal degeneration and in patients suffering these diseases. The goal of our research is to improve current procedures to the stage in which they can be used routinely to alleviate these causes of blindness in people.

We are differentiating human embryonic stem cells (hESCs) into sheets of retinal progenitor tissue that contain photoreceptor progenitors and RPE cells. We anticipate that the successful completion of our research will create an unlimited supply of donor tissue that can be transplanted into the eyes of blind people. Recently, several laboratories have shown that hESCs and induced pluripotent stem cells (iPSCs) can “self-assemble” into early stages of eye development and develop into laminated structures resembling the normal retina. The hypothesis of this project is that hESCs can be consistently differentiated into sheets of retinal tissue, which can restore visual responses in an immunodeficient rat model of retinal degeneration that does not reject human cells. In the third year of our research program, we will standardize methods to increase the production of these sheets in a way that complies with good manufacturing practice. This project will ultimately help to restore vision in patients suffering from retinal diseases.

In year 1 of this 3-year grant, the optimal procedure for differentiation of hESCs into laminated 3D-retina has been developed and tested. This method is based on selection of tissues developing into retinal progenitor cells, and culturing them in suspension (non-attached). We have shown by quantitative PCR and by immunohistochemistry that these tissues express retina-specific genes in a time sequence comparable to human retinal development. We have built up the capabilities for testing hESC-derived 3D retina in vivo. We have set up a colony of immunodeficient retinal degenerate (RD) rats (which do not require immunosuppression), and refined the transplantation, imaging and testing techniques. To hone our skills, we have started with transplants of rat and human fetal retinal tissue. We will start with transplantation of hESC-derived 3D retina in March 2015 and continue with testing the tissue through the remainder of the grant. The development of the transplants in vivo is monitored by high-resolution optical coherence tomography which can image retinal layers. Transplanted RD rats are tested by various methods (optokinetic tracking for visual acuity, electroretinograms and recording of visual responses from the superior colliculs, a visual brain center) to demonstrate the transplant effect on vision. In the first year of the grant, we have shown that these methods can demonstrate the improvement of vision by fetal-derived transplants

This work will be presented in several posters at this year’s ARVO meeting; several manuscripts are in preparation. The laboratory has attracted many students who are interested to learn about retinal development and vision restoration.