GMP-Grade Manufacturing and Quality Control of a Non-Virally Engineered Advanced Therapy Medicinal Product for Personalized Treatment of Age-Related Macular Degeneration

Abstract

The introduction of new therapeutics requires validation of Good Manufacturing Practice (GMP)-grade manufacturing including suitable quality controls. This is challenging for Advanced Therapy Medicinal Products (ATMP) with personalized batches. We have developed a person-alized, cell-based gene therapy to treat age-related macular degeneration and established a vali-dation strategy of the GMP-grade manufacture for the ATMP; manufacturing and quality control were challenging due to a low cell number, batch-to-batch variability and short production duration. Instead of patient iris pigment epithelial cells, human donor tissue was used to produce the transfected cell product (“tIPE”). We implemented an extended validation of 104 tIPE productions. Procedure, operators and devices have been validated and qualified by determining cell number, viability, extracellular DNA, sterility, duration, temperature and volume. Transfected autologous cells were transplanted to rabbits verifying feasibility of the treatment. A container has been engineered to ensure a safe transport from the production to the surgery site. Criteria for successful validation and qualification were based on tIPE’s Critical Quality Attributes and Process Parameters, its manufacture and release criteria. The validated process and qualified operators are essential to bring the ATMP into clinic and offer a general strategy for the transfer to other manufacture centers and personalized ATMPs.

Mammalian Animal and Human Retinal Organ Culture as Pre-Clinical Model to Evaluate Oxidative Stress and Antioxidant Intraocular Therapeutics

“…Model 4 has been implemented to prolong the culture, and its avail was evaluated by analyzing the effect of PEDF and/or GM-CSF on cell viability and OS after the H 2 O 2 treatment. The rat retinae were cultured immediately after their sacrifice, which allowed cultures up to 13 d and confirmed the cell viability rates in freshly fixed retinae to be around 31%, which has also been measured in IPE cells immediately after isolation [ 62 ]. After the H 2 O 2 treatment, reduced GSH levels were expected as reported by Wang et al in ARPE-19 cells [ 63 ] and by Zheng et al in retinae from 18-month-old hydroquinone-fed mice [ 64 ].…”

Antibiotic-Free Gene Vectors: A 25-Year Journey to Clinical Trials

“…A battery of preclinical assays has been performed [ 94 , 95 , 96 , 97 , 98 , 112 ], including in vitro IPE cells’ transfection, a quantification of the secreted PEDF level and the assessment of transgene integration sites which were, as expected with the SB transposon system, close to random (see Section 7 below on transposons). Furthermore, the transplantation of the genetically modified cells to the subretinal space of a rat model significantly reduced the laser-induced choroidal neovascularization and did not generate any change in rabbit organ morphologies (Biodistribution studies).…”

Enhanced Biosafety of the Sleeping Beauty Transposon System by Using mRNA as Source of Transposase to Efficiently and Stably Transfect Retinal Pigment Epithelial Cells

“…VEGF-induced sprouting and migration of human umbilical vein endothelial cells (HUVEC) was inhibited by rPEDF with a 2000 times higher potency compared with bevacizumab [ 21 ]. We recently established the GMP-compliant production of PEDF-overexpressing IPE cells to evaluate the safety of this treatment approach in a first-in-human phase 1b/2a clinical trial [ 25 ].…”