Discovery of Novel Cell Surface Markers for Purification of Embryonic Dopamine Progenitors for Transplantation in Parkinson's Disease Animal Models

• Published in: Molecular & cellular proteomics 2018-09, Vol.17 (9), p.1670-1684
• Main Authors: Fathi, Ali, Mirzaei, Mehdi, Dolatyar, Banafsheh, Sharifitabar, Mehdi, Bayat, Mahnaz, Shahbazi, Ebrahim, Lee, Jaesuk, Javan, Mohammad, Zhang, Su-Chun, Gupta, Vivek, Lee, Bonghee, Haynes, Paul A., Baharvand, Hossein, Salekdeh, Ghasem Hosseini
• Format: Article
• Language: English
• Subjects: Animals; Biomarkers - metabolism; Cell Differentiation; Cell Membrane - metabolism; Cell Separation - methods; Contactin 2 - metabolism; Disease Models, Animal; Dopaminergic Neurons - transplantation; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Female; Green Fluorescent Proteins - metabolism; Humans; Label-free quantification; LIM-Homeodomain Proteins - metabolism; Parkinson Disease - pathology; Parkinson Disease - therapy; Parkinson's disease; Progenitor cells; Proteomics; Rats, Sprague-Dawley; Reproducibility of Results; Spectral counting; Stem cells; Transcription Factors - metabolism
• ISSN: 1535-9476; 1535-9484
• DOI: 10.1074/mcp.RA118.000809

Despite the progress in safety and efficacy of cell replacement therapy with pluripotent stem cells (PSCs), the presence of residual undifferentiated stem cells or proliferating neural progenitor cells with rostral identity remains a major challenge. Here we report the generation of a LIM homeobox transcription factor 1 alpha (LMX1A) knock-in GFP reporter human embryonic stem cell (hESC) line that marks the early dopaminergic progenitors during neural differentiation to find reliable membrane protein markers for isolation of midbrain dopaminergic neurons. Purified GFP positive cells in vitro exhibited expression of mRNA and proteins that characterized and matched the midbrain dopaminergic identity. Further quantitative proteomics analysis of enriched LMX1A+ cells identified several membrane-associated proteins including a polysialylated embryonic form of neural cell adhesion molecule (PSA-NCAM) and contactin 2 (CNTN2), enabling prospective isolation of LMX1A+ progenitor cells. Transplantation of human-PSC-derived purified CNTN2+ progenitors enhanced dopamine release from transplanted cells in the host brain and alleviated Parkinson's disease-related phenotypes in animal models. This study establishes an efficient approach for purification of large numbers of human-PSC-derived dopaminergic progenitors for therapeutic applications.