Transcriptome analysis reveals transmembrane targets on transplantable midbrain dopamine progenitors

Proc Natl Acad Sci U S A. 2015 Apr 14;112(15):E1946-55. doi: 10.1073/pnas.1501989112. Epub 2015 Mar 9.

Abstract

An important challenge for the continued development of cell therapy for Parkinson's disease (PD) is the establishment of procedures that better standardize cell preparations for use in transplantation. Although cell sorting has been an anticipated strategy, its application has been limited by lack of knowledge regarding transmembrane proteins that can be used to target and isolate progenitors for midbrain dopamine (mDA) neurons. We used a "FACS-array" approach to identify 18 genes for transmembrane proteins with high expression in mDA progenitors and describe the utility of four of these targets (Alcam, Chl1, Gfra1, and Igsf8) for isolating mDA progenitors from rat primary ventral mesencephalon through flow cytometry. Alcam and Chl1 facilitated a significant enrichment of mDA neurons following transplantation, while targeting of Gfra1 allowed for robust separation of dopamine and serotonin neurons. Importantly, we also show that mDA progenitors isolated on the basis of transmembrane proteins are capable of extensive, functional innervation of the host striatum and correction of motor impairment in a unilateral model of PD. These results are highly relevant for current efforts to establish safe and effective stem cell-based procedures for PD, where clinical translation will almost certainly require safety and standardization measures in order to deliver well-characterized cell preparations.

Keywords: Alcam; Parkinson’s disease; cell sorting; microarray; transplantation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activated-Leukocyte Cell Adhesion Molecule / genetics
  • Activated-Leukocyte Cell Adhesion Molecule / metabolism
  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / metabolism
  • Cells, Cultured
  • Dopaminergic Neurons / cytology
  • Dopaminergic Neurons / metabolism*
  • Dopaminergic Neurons / transplantation
  • Female
  • Flow Cytometry / methods
  • Gene Expression Profiling*
  • Gene Expression Regulation, Developmental
  • Glial Cell Line-Derived Neurotrophic Factor Receptors / genetics
  • Glial Cell Line-Derived Neurotrophic Factor Receptors / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • LIM-Homeodomain Proteins / genetics
  • LIM-Homeodomain Proteins / metabolism
  • Male
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mesencephalon / cytology
  • Mesencephalon / embryology
  • Mesencephalon / metabolism
  • Mice, Inbred Strains
  • Mice, Transgenic
  • Microscopy, Confocal
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism*
  • Neural Stem Cells / transplantation
  • Parkinson Disease / therapy
  • Rats, Sprague-Dawley
  • Stem Cell Transplantation / methods*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Activated-Leukocyte Cell Adhesion Molecule
  • Basic Helix-Loop-Helix Transcription Factors
  • Carrier Proteins
  • Cell Adhesion Molecules
  • Chl1 protein, mouse
  • Gfra1 protein, mouse
  • Glial Cell Line-Derived Neurotrophic Factor Receptors
  • IgSF8 protein, mouse
  • LIM-Homeodomain Proteins
  • Lmx1a protein, mouse
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Neurog2 protein, mouse
  • Transcription Factors
  • Green Fluorescent Proteins

Associated data

  • GEO/GSE65094