title: Imaging Flow Cytometry Analysis to Identify Differences of Survival Motor Neuron Protein Expression in Patients With Spinal Muscular Atrophy.
abstract: Spinal muscular atrophy is a neurodegenerative disorder caused by the deficient expression of survival motor neuron protein in motor neurons. A major goal of disease-modifying therapy is to increase survival motor neuron expression. Changes in survival motor neuron protein expression can be monitored via peripheral blood cells in patients; therefore we tested the sensitivity and utility of imaging flow cytometry for this purpose.
  
  
  
  After the immortalization of peripheral blood lymphocytes from a human healthy control subject and two patients with spinal muscular atrophy type 1 with two and three copies of SMN2 gene, respectively, we used imaging flow cytometry analysis to identify significant differences in survival motor neuron expression. A bright detail intensity analysis was used to investigate differences in the cellular localization of survival motor neuron protein.
  
  
  
  Survival motor neuron expression was significantly decreased in cells derived from patients with spinal muscular atrophy relative to those derived from a healthy control subject. Moreover, survival motor neuron expression correlated with the clinical severity of spinal muscular atrophy according to SMN2 copy number. The cellular accumulation of survival motor neuron protein was also significantly decreased in cells derived from patients with spinal muscular atrophy relative to those derived from a healthy control subject.
  
  
  
  The benefits of imaging flow cytometry for peripheral blood analysis include its capacities for analyzing heterogeneous cell populations; visualizing cell morphology; and evaluating the accumulation, localization, and expression of a target protein. Imaging flow cytometry analysis should be implemented in future studies to optimize its application as a tool for spinal muscular atrophy clinical trials.
authors.affiliation: Institute of Medical Genetics, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Arakawa
authors.affiliation: Laboratory of Virology, Institute of Microbial Chemistry, Microbial Chemistry Research Foundation, Tokyo, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Arakawa
authors.affiliation: Institute of Medical Genetics, Tokyo Women's Medical University, Tokyo, Japan; Affiliated Field of Medical Genetics, Division of Biomedical Engineering and Science, Graduate Course of Medicine, Graduate School of Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Kaneko
authors.affiliation: Institute of Medical Genetics, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Otsuki
authors.affiliation: Institute of Medical Genetics, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Ryoko
authors.initials: R
authors.lastname: Aoki
authors.affiliation: Institute of Medical Genetics, Tokyo Women's Medical University, Tokyo, Japan; Affiliated Field of Medical Genetics, Division of Biomedical Engineering and Science, Graduate Course of Medicine, Graduate School of Tokyo Women's Medical University, Tokyo, Japan. Electronic address: saito.kayoko@twmu.ac.jp.
authors.forename: Kayoko
authors.initials: K
authors.lastname: Saito
mesh_terms: D002675:Child, Preschool; D056915:DNA Copy Number Variations; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D008214:Lymphocytes / Q000378:metabolism / Q000473:pathology; D008297:Male; D009134:Muscular Atrophy, Spinal / Q000378:metabolism* / Q000473:pathology; D055533:Survival of Motor Neuron 1 Protein / Q000235:genetics / Q000378:metabolism; D055540:Survival of Motor Neuron 2 Protein / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome
chemical_list: C526471:SMN1 protein, human; C526477:SMN2 protein, human; D055533:Survival of Motor Neuron 1 Protein; D055540:Survival of Motor Neuron 2 Protein
keywords: imaging flow cytometry; peripheral blood analysis; spinal muscular atrophy; survival motor neuron protein