KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Induction of Midbrain Dopaminergic Phenotype in Nurr 1-Over expressing Human Neural Stem Cells

사람 신경 간세포에서 도파민 신경세포 분화유도에 대한 Nurr 1 유전자의 역할 규명

  • Kim, Han-Jip (Department of Biology, College of Natural Science, Ajou University) ;
  • Lee, Haksup (Department of Biology, College of Natural Science, Ajou University) ;
  • Kim, Hyon-Chang (Department of Biology, College of Natural Science, Ajou University) ;
  • Min, Churl-Ki (Department of Biology, College of Natural Science, Ajou University) ;
  • Lee, Myung-Ae (Brain Disease Research Center, College of Medicine, Ajou University) ;
  • Kim, Seung-Up (Brain Disease Research Center, College of Medicine, Ajou University) ;
  • Han, Jin (Mitochondrial Signaling Laboratory, Department of Molecular Physiology & Biophysics, College of Medicine, Inje University) ;
  • Youm, Jae-Boum (Mitochondrial Signaling Laboratory, Department of Molecular Physiology & Biophysics, College of Medicine, Inje University) ;
  • Kim, Nari (Mitochondrial Signaling Laboratory, Department of Molecular Physiology & Biophysics, College of Medicine, Inje University) ;
  • Park, Won, Sun (Mitochondrial Signaling Laboratory, Department of Molecular Physiology & Biophysics, College of Medicine, Inje University) ;
  • Kim, Taeho (Mitochondrial Signaling Laboratory, Department of Molecular Physiology & Biophysics, College of Medicine, Inje University) ;
  • Kim, Euiyong (Mitochondrial Signaling Laboratory, Department of Molecular Physiology & Biophysics, College of Medicine, Inje University) ;
  • Han, Il-Yong (Department of Thoracic & Cardiovascular Surgery, College of Medicine, Inje University)
  • 김한집 (아주대학교 자연과학대학 생명과학전공) ;
  • 이학섭 (아주대학교 자연과학대학 생명과학전공) ;
  • 김현창 (아주대학교 자연과학대학 생명과학전공) ;
  • 민철기 (아주대학교 자연과학대학 생명과학전공) ;
  • 이명애 (아주대학교 의과대학 뇌질환연구소) ;
  • 김승업 (아주대학교 의과대학 뇌질환연구소) ;
  • 한진 (인제대학교 의과대학 미토콘드리아 생체신호 연구실, 생리학교실) ;
  • 염재범 (인제대학교 의과대학 미토콘드리아 생체신호 연구실, 생리학교실) ;
  • 김나리 (인제대학교 의과대학 미토콘드리아 생체신호 연구실, 생리학교실) ;
  • 박원선 (인제대학교 의과대학 미토콘드리아 생체신호 연구실, 생리학교실) ;
  • 김태호 (인제대학교 의과대학 미토콘드리아 생체신호 연구실, 생리학교실) ;
  • 김의용 (인제대학교 의과대학 미토콘드리아 생체신호 연구실, 생리학교실) ;
  • 한일용 (인제대학교 의과대학 부산백병원 흉부외과학교실)
  • Published : 2005.10.01
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Abstract

Neural stem cells (NSCs) of the central nervous system (CNS) have raised a great interest not only for their importance in basic neural development but also for their therapeutic potentials in neurologically degenerative diseases such as Parkinson's, Alzheimer and stroke. During the CNS development, two molecular cascades determine specification of midbrain dopamine system. In one pathway, FGF-8, sonic hedgehog and transcription factor Nurr1 specify dopamine neurotransmitter phenotype. In the other, transcription factors $Lm{\times}lb\;and\;Pt{\times}3$ are required for induction of dopaminergic neurons. In Nurr1 knockout mouse, tyrosine hydroxylase (TH) positive cells fail to appear in substantia nigra, indicating that Nurr1 is essential in specification of dopaminergic cell phenotype. In this study, we used the immortalized human NSCs retrovirally transduced with Nurr1 gene to probe the Nurr1 mediated mechanism to induce dopamine phenotype. While Nurr1 over-expression alone did not generate dopamine phenotype in NSCs, applications of retinoid and forskolin induced expression of TH and AADC mRNAs. In addition, co-cultures of Nurr1 expressing NSCs with human astrocytes induced a marked increase of TH expression. In this co-culture system, the addition of retinoid and forskolin dramatically increased expression of TH. These results indicate that the immortalized human NSCs with Nurr1 gene could have a clinical utility for cell replacement for the Parkinson patients.

중추신경계의 신경간세포가 파킨슨병과 뇌졸중과 같은 퇴행성 뇌 질환의 치료뿐만이 아니라 신경세포 발생과정에서의 중요성 때문에 최근에 커다란 관심의 대상이 되고 있다. 중추신경계의 발생과정 동안에, 중뇌의 도파민 신경세포의 형성은 두 가지의 분자생물학적인 기작에 의해서 결정된다. 첫째로, FGF-8, sonic hedgehog 그리고 전사조절인자 인 Nurr1이 도파민 신경세포의 형질을 결정짓는다. 또 다른 기작으로는, 전사조절인자 인 $Lm{\times}lb$와 $Pt{\times}3$가 중요하게 관련되어있다. 특히 Nurr1이 결핍된 생쥐에서, 타이로신수산화효소 (Tyrosine bydroxylase, TH) 면역양성 세포들이 중뇌흑색질에서 발견되지 않으므로 Nurr1이 도파민 신경세포의 발생에 필수적임을 알 수 있다. 본 연구에서는 도파민 신경세포의 형질을 유도하는데 있어서 Nurr1이 매개하는 기작을 연구하기 위해서 레트로 바이러스를 이용하여 Nurr1을 도입한 무한증식 신경간세포를 사용하였다. Nurr1 유전자의 과발현 만으로는 신경간세포에서 도파민 신경세포의 형질을 유도하지는 못하지만, 레티노이드 (retinoid, RA)와 폴스콜린 (forskolin, FK)을 처리하여 TH와 방향성 L-아미노산 탈카르복시화효소 (aromatic L-amino acid decarboxylase, AADC) mRNA의 발현을 유도하였다. 또한, Nurr1 과발현 신경간세포를 사람 별아교세포와 공동배양 하여 TH 발현량을 많이 증가시켰다. 이러한 공동배양실험에서, RA와 FK를 처리하면 TH의 발현수준이 더욱 더 증가함을 발견하였다. 이러한 결과들은 Nurr1 유전자를 도입한 사람 신경간세포가 파킨슨병 환자들에게 세포이식을 통한 유전자 치료의 유용성을 시사하고 있다.

Keywords

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