Discovery of novel cell surface markers for purification of embryonic dopamine progenitors for transplantation in Parkinson's disease animal models

Ali Fathi, Mehdi Mirzaei, Banafsheh Dolatyar, Mehdi Sharifitabar, Mahnaz Bayat, Ebrahim Shahbazi, Jaesuk Lee, Mohammad Javan, Su-Chun Zhang, Vivek Gupta, Bonghee Lee, Paul A. Haynes, Hossein Baharvand, Ghasem Hosseini Salekdeh

Research output: Contribution to journalArticleResearchpeer-review

Abstract

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.

LanguageEnglish
Pages1670-1684
Number of pages15
JournalMolecular and Cellular Proteomics
Volume17
Issue number9
DOIs
Publication statusPublished - 1 Sep 2018

Fingerprint

Pluripotent Stem Cells
Contactin 2
Stem cells
Purification
Parkinson Disease
Dopamine
Animals
Stem Cells
Animal Models
Transplantation
Mesencephalon
Membrane Proteins
Neural Cell Adhesion Molecules
Homeobox Genes
Dopaminergic Neurons
Cell- and Tissue-Based Therapy
Proteomics
Transcription Factors
Phenotype
Safety

Cite this

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. / Discovery of novel cell surface markers for purification of embryonic dopamine progenitors for transplantation in Parkinson's disease animal models. In: Molecular and Cellular Proteomics. 2018 ; Vol. 17, No. 9. pp. 1670-1684.
@article{df5c4da979414b0b988d35cdc0c0b9e2,
title = "Discovery of novel cell surface markers for purification of embryonic dopamine progenitors for transplantation in Parkinson's disease animal models",
abstract = "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.",
author = "Ali Fathi and Mehdi Mirzaei and Banafsheh Dolatyar and Mehdi Sharifitabar and Mahnaz Bayat and Ebrahim Shahbazi and Jaesuk Lee and Mohammad Javan and Su-Chun Zhang and Vivek Gupta and Bonghee Lee and Haynes, {Paul A.} and Hossein Baharvand and Salekdeh, {Ghasem Hosseini}",
year = "2018",
month = "9",
day = "1",
doi = "10.1074/mcp.RA118.000809",
language = "English",
volume = "17",
pages = "1670--1684",
journal = "Molecular & cellular proteomics : MCP",
issn = "1535-9476",
publisher = "American Society for Biochemistry and Molecular Biology",
number = "9",

}

Discovery of novel cell surface markers for purification of embryonic dopamine progenitors for transplantation in Parkinson's disease animal models. / 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.

In: Molecular and Cellular Proteomics, Vol. 17, No. 9, 01.09.2018, p. 1670-1684.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Discovery of novel cell surface markers for purification of embryonic dopamine progenitors for transplantation in Parkinson's disease animal models

AU - Fathi,Ali

AU - Mirzaei,Mehdi

AU - Dolatyar,Banafsheh

AU - Sharifitabar,Mehdi

AU - Bayat,Mahnaz

AU - Shahbazi,Ebrahim

AU - Lee,Jaesuk

AU - Javan,Mohammad

AU - Zhang,Su-Chun

AU - Gupta,Vivek

AU - Lee,Bonghee

AU - Haynes,Paul A.

AU - Baharvand,Hossein

AU - Salekdeh,Ghasem Hosseini

PY - 2018/9/1

Y1 - 2018/9/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85052728954&partnerID=8YFLogxK

U2 - 10.1074/mcp.RA118.000809

DO - 10.1074/mcp.RA118.000809

M3 - Article

VL - 17

SP - 1670

EP - 1684

JO - Molecular & cellular proteomics : MCP

T2 - Molecular & cellular proteomics : MCP

JF - Molecular & cellular proteomics : MCP

SN - 1535-9476

IS - 9

ER -