Director, Analytical Development Aspen Neuroscience Inc., United States
Abstract: While the potential of cell replacement therapy for Parkinson’s Disease was first demonstrated decades ago, extensive work was performed since then to develop stem cell derived neural progenitor cells with the optimal phenotype for an efficacious drug product. An additional challenge is to sufficiently characterize the potency of these progenitor cells that require in vivo maturation to perform functions critical to their mechanism of action.
A novel suite of tests was developed to address these challenges. As Aspen Neuroscience advances an autologous iPSC-derived cell replacement therapy for Parkinson’s Disease into clinical trials, we built a suite of analytical methods to serve as a potency assay matrix of iPSC-derived dopaminergic neuron precursor cells (DANPCs) product.
Here, we show that a set of bioinformatics tests performed on bulk RNA sequencing (RNAseq) data of DANPCs, an analytical matrix which provides predictive analytics for the optimal maturity/identity, engraftment potential, and dopamine production potential. After utilizing a large, high-quality training dataset, these bioinformatics tests can reliably predict in vivo maturation, engraftment efficiency and dopamine production of DANPCs post transplantation. To overcome the limitation of bulk RNAseq not being able to capture the biochemical or physical function of DANPCs post-thaw, an enzymatic assay has been established to evaluate DANPCs functionality post-thaw. Dopa decarboxylase (DDC) was identified as an enzyme that is critical to the production of dopamine and is highly expressed in DANPCs. A DDC enzymatic activity assay was developed to assess the post-thaw function of DANPCs.
Establishing the potency of stem cell derived therapies remains a challenge for many potential products, and novel work is required to address this early in the development of a therapy. This stability-indicating, conventional, enzymatic bioassay of DANPCs functionality in combination with the predictive bulk RNAseq-based bioinformatics tests provides a robust potency assay matrix strategy for Parkinson’s Disease cell replacement therapy.
Funding Source: Aspen Neuroscience Inc, CIRM, Summit for Stem Cell
