Transplantation of post-mitotic human neuroteratocarcinoma-overexpressing Nurr1 cells provides therapeutic benefits in experimental stroke: in vitro evidence of expedited neuronal differentiation and GDNF secretion.
Nurr1 has been implicated as a transcription factor mediating the endogenous neuroprotective mechanism against stroke. We examined the in vivo and in vitro properties of a new human embryonic carcinoma Ntera-2 cell line carrying the human Nurr1 gene (NT2N.Nurr1). Adult Sprague-Dawley rats underwent experimental stroke initially and 14 days later were assigned randomly to receive stereotaxic transplantation of NT2N.Nurr1 cells or infusion of vehicle into their ischemic striatum. Transplantation of NT2N.Nurr1 cells promoted significant attenuation of behavioral impairments over a 56-day period after stroke, characterized by decreased hyperactivity, biased swing activity, and neurologic deficits, as well as significant reduction in ischemic striatal cell loss compared to vehicle-infused stroke animals. Transplanted NT2N.Nurr1 cells survived and expressed neuronal phenotypic markers in the ischemic striatum. In vitro results showed that cultured NT2.Nurr1 cells were already negative for nestin even before retinoic acid treatment, despite strong nestin immunoreactivity in NT2 cells. This indicates Nurr1 triggered a rapid commitment of NT2 cells into a neuronal lineage. Indeed, NT2.Nurr1 cells, at 4 weeks into RA treatment, displayed more abundant tyrosine hydroxylase positive cells than NT2 cells. Parallel ELISA studies showed further that cultured NT2N.Nurr1, but not NT2N cells, secreted glial cell derived neurotrophic factor. The present study shows efficacy of NT2N.Nurr1 cell grafts in ischemic stroke, with in vitro evidence suggesting the cells' excellent neuronal differentiation capability and ability to secrete GDNF as likely mechanisms mediating the observed therapeutic benefits.
Hara
Koichi
K
Department of Neurology, Medical College of Georgia, Augusta, GA 30912, USA.
Matsukawa
Noriyuki
N
Yasuhara
Takao
T
Xu
Lin
L
Yu
Guolong
G
Maki
Mina
M
Kawase
Takeshi
T
Hess
David C
DC
Kim
Seung U
SU
Borlongan
Cesar V
CV
D000818:Animals; D001522:Behavior, Animal; D002277:Carcinoma / Q000473:pathology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D045744:Cell Line, Tumor; D017690:Cell Transplantation / Q000379:methods*; D003342:Corpus Striatum / Q000378:metabolism / Q000601:surgery; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000627:therapeutic use*; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism*; D006801:Humans; D008297:Male; D009043:Motor Activity / Q000502:physiology; D015999:Multivariate Analysis; D009419:Nerve Tissue Proteins / Q000378:metabolism; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D051381:Rats; D017207:Rats, Sprague-Dawley; D020521:Stroke / Q000473:pathology / Q000601:surgery*; D013997:Time Factors; D014157:Transcription Factors / Q000096:biosynthesis / Q000627:therapeutic use*; D014212:Tretinoin / Q000494:pharmacology
D004268:DNA-Binding Proteins; D051100:Glial Cell Line-Derived Neurotrophic Factor; C539882:NR4A2 protein, human; D009419:Nerve Tissue Proteins; C539884:Nr4a2 protein, rat; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D014157:Transcription Factors; D014212:Tretinoin