RPE4AMD

About the project:

The project aims to develop a standardized, safe and efficient method for curing dry age-related macular degeneration (AMD) – the most frequent cause of visual impairment that currently lacks treatment. The work will bring promising retina pigment epithelial (RPE) cell research to a clinical trial using patient-derived stem cell therapy. The Specific aims are focused at: 1) isolation, cultivation and optimization of human RPE cells, primary hRPEs and induced pluripotent stem cell (iPSC)-derived RPEs; 2) preparation of nanofibrous scaffold; 3) transplantation of hRPE-covered scaffolds in the subretinal space; 4) non-invasive (OCT, mfERG) and end-stage cell and tissue characterization (IHC, tEM); 5) establishment of good manufacturing practice (GMP) protocols and clinical-grade cell banking.

2021

We successfully optimized isolation and cultivation protocol for RPE cells with high efficiency to form a monolayered epithelium. We performed many examinations of the growing and maturation of culturing porcine RPE on the standard commercial cell culture porous inserts and our nanofibrous membranes. Using various examination methods, we found good conditions for cell growing on both membranes but with better results of RPE markers on nanofibrous membranes in comparison with commercial ones. For evaluation of the epithelium quality respective tight junctions, we measured an epithelial electrical resistance where the resulting values demonstrated well cell health.

2022

In the year 2022, the new method for culture of the human induced pluripotent cells and their differentiation in the human retina pigment epithelial (RPE) cells was introduced. This new approach was done by Mgr. Anna Brymova in Dr. Slaven Erceg’s Laboratory in Valencia, Spain. The human RPE cultured on the nanofibrous membranes, prepared by Dr. Hana Studenovska, IMCH, CAS, were transferred in the subretinal space of the immunosuppressed minipigs. This experiment represents an important step on the way to the AMD cell therapy.

2023

All teams involved did during the third year the substantial progress in their methodologies. IAPG CAS in Libechov tuned, in cooperation with BIOINOVA Company, methods of culture and differentiation of the human iPS cells in the pigmented retinal (RPE) cells. The team at IMC, CAS prepared and defended the methodology of repeatable preparation of the nanofibrous membranes. Banking of the nanofibrous membranes with RPE cells has been initiated and it is in progress now. Development of the precise method of immunosuppression represents precondition for the successful implementation in the subretinal space of recipient minipigs.

Summary

The project represents the unique example of Czech-Norwegian cooperation in the biomedical research. The key mission of these experiments was to develop the standardized methods for culture of retina pigmented epithelial cells (RPE). RPE cells are primarily affected at age-related macular degeneration (AMD). Degeneration of RPE cells results in a loss of photoreceptors and gradual impairment of vision. The great methodological progress in preparation of induced pluripotent stem cells (iPS) enabled, in a broad international cooperation, to develop the method of differentiation of iPS cells in the human retinal pigment epithelial cells. RPE cells are cultured on the nanofibrous membranes. The prepared “sandwich” is implanted under retina of Libechov minipigs. We verified this way, safety and efficiency of all newly developed methods. These experiments are key to entering the clinic, for the future treatment of age-related macular degeneration using cell therapy.

Beneficiary and project partners:

Ústav živočišné fyziologie a genetiky AV ČR, v. v. i.
Fakultní nemocnice Královské Vinohrady
University of Oslo
Ústav makromolekulární chemie AV ČR, v. v. i.

The RPE4AMD project benefits from a € 1 mil. grant from Norway Grants and Technology Agency of the Czech Republic. The project is carried out under the KAPPA funding programme for applied research, experimental development and innovation, managed by the Technology Agency of the Czech Republic.