Mitochondrial Dysfunction of Immortalized Human Adipose Tissue-Derived Mesenchymal Stromal Cells from Patients with Parkinson’s Disease

다국어 초록 (Multilingual Abstract)

Mitochondrial dysfunction in dopaminergic neurons of patients with idiopathic and familial Parkinson’s disease (PD) is well knownalthough the underlying mechanism is not clear. We established a homogeneous population of human adipose tissue-derivedmesenchymal stromal cells (hAD-MSCs) from human adult patients with early-onset hereditary familial Parkin-defect PD as wellas late-onset idiopathic PD by immortalizing cells with the hTERT gene to better understand the underlying mechanism of PD.
The hAD-MSCs from patients with idiopathic PD were designated as “PD”, from patients with Parkin-defect PD as “Parkin” andfrom patients with pituitary adenomas as “non-PD” in short. The pGRN145 plasmid containing hTERT was introduced to establishtelomerase immortalized cells. The established hTERT-immortalized cell lines showed chromosomal aneuploidy sustained stably overtwo-years. The morphological study of mitochondria in the primary and immortalized hAD-MSCs showed that the mitochondriaof the non-PD were normal; however, those of the PD and Parkin were gradually damaged. A striking decrease in mitochondrialcomplex I, II, and IV activities was observed in the hTERT-immortalized cells from the patients with idiopathic and Parkin-defectPD. Comparative Western blot analyses were performed to investigate the expressions of PD specific marker proteins in the hTERT-immortalized cell lines. This study suggests that the hTERT-immortalized hAD-MSC cell lines established from patients withidiopathic and familial Parkin-defect PD could be good cellular models to evaluate mitochondrial dysfunction to better understandthe pathogenesis of PD and to develop early diagnostic markers and effective therapy targets for the treatment of PD.

참고문헌 (Reference)

1 Lee KM, "Use of exogenous hTERT to immortalize primary human cells" 45 : 33-38, 2004

2 Schenkman JB, "The many roles of cytochrome b5" 97 : 139-152, 2003

3 Morin GB, "The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats" 59 : 521-529, 1989

4 Peng J, "The herbicide paraquat induces dopaminergic nigral apoptosis through sustained activation of the JNK pathway" 279 : 32626-32632, 2004

5 Shim JH, "The antioxidant Trolox helps recovery from the familial Parkinson’s diseasespecific mitochondrial deficits caused by PINK1- and DJ-1- deficiency in dopaminergic neuronal cells" 11 : 707-715, 2011

6 Wen VW, "Telomere-driven karyotypic complexity concurs with p16INK4a inactivation in TP53-competent immortal endothelial cells" 66 : 10691-10700, 2006

7 Belgiovine C, "Telomerase: cellular immortalization and neoplastic transformation. Multiple functions of a multifaceted complex" 122 : 255-262, 2008

8 Kuan CY, "Targeting the JNK signaling pathway for stroke and Parkinson’s diseases therapy" 4 : 63-67, 2005

9 Schapira AH, "Targeting mitochondria for neuroprotection in Parkinson’s disease" 16 : 965-973, 2012

10 Blackburn EH, "Structure and function of telomeres" 350 : 569-573, 1991

11 Janssen AJ, "Spectrophotometric assay for complex I of the respiratory chain in tissue samples and cultured fibroblasts" 53 : 729-734, 2007

12 Engelhardt JF, "Redox-mediated gene therapies for environmental injury: approaches and concepts" 1 : 5-27, 1999

13 Hoepken HH, "Parkinson patient fibroblasts show increased alpha-synuclein expression" 212 : 307-313, 2008

14 Fauth C, "Order of genetic events is critical determinant of aberrations in chromosome count and structure" 40 : 298-306, 2004

15 Pittenger MF, "Multilineage potential of adult human mesenchymal stem cells" 284 : 143-147, 1999

16 Martin J, "Molecular chaperones and mitochondrial protein folding" 29 : 35-43, 1997

17 Greene JC, "Mitochondrial pathology and apoptotic muscle degeneration in Drosophila parkin mutants" 100 : 4078-4083, 2003

18 Palacino JJ, "Mitochondrial dysfunction and oxidative damage in parkin-deficient mice" 279 : 18614-18622, 2004

19 Müftüoglu M, "Mitochondrial complex I and IV activities in leukocytes from patients with parkin mutations" 19 : 544-548, 2004

20 Henchcliffe C, "Mitochondrial biology and oxidative stress in Parkinson disease pathogenesis" 4 : 600-609, 2008

21 Buckman JF, "MitoTracker labeling in primary neuronal and astrocytic cultures: influence of mitochondrial membrane potential and oxidants" 104 : 165-176, 2001

22 Hahn WC, "Inhibition of telomerase limits the growth of human cancer cells" 5 : 1164-1170, 1999

23 Dexter DT, "Increased nigral iron content and alterations in other metal ions occurring in brain in Parkinson’s disease" 52 : 1830-1836, 1989

24 Greider CW, "Identification of a specific telomere terminal transferase activity in Tetrahymena extracts" 43 : 405-413, 1985

25 Zuk PA, "Human adipose tissue is a source of multipotent stem cells" 13 : 4279-4295, 2002

26 Gorbatyuk MS, "Glucose regulated protein 78 diminishes α-synuclein neurotoxicity in a rat model of Parkinson disease" 20 : 1327-1337, 2012

27 문효은, "Genetic Profiling in Human Adipose Tissue-Derived Mesenchymal Stromal Cells from the Idiopathic and Familial Parkin-Deficient Patients of Parkinson's Disease in Comparison with non-PD patients" 한국조직공학과 재생의학회 7 (7): 237-247, 2010

28 Bodnar AG, "Extension of life-span by introduction of telomerase into normal human cells" 279 : 349-352, 1998

29 Stefanis L, "Expression of A53T mutant but not wild-type alphasynuclein in PC12 cells induces alterations of the ubiquitindependent degradation system, loss of dopamine release, and autophagic cell death" 21 : 9549-9560, 2001

30 Yoo MS, "Experimental strategy to identify genes susceptible to oxidative stress in nigral dopaminergic neurons" 29 : 1223-1234, 2004

31 Abou-Sleiman PM, "Expanding insights of mitochondrial dysfunction in Parkinson’s disease" 7 : 207-219, 2006

32 Sriram K, "Evidence for generation of oxidative stress in brain by MPTP: in vitro and in vivo studies in mice" 749 : 44-52, 1997

33 Tsuruga Y, "Establishment of immortalized human hepatocytes by introduction of HPV16 E6/E7 and hTERT as cell sources for liver cell-based therapy" 17 : 1083-1094, 2008

34 Kehat I, "Electromechanical integration of cardiomyocytes derived from human embryonic stem cells" 22 : 1282-1289, 2004

35 Uryu K, "Convergence of heat shock protein 90 with ubiquitin in filamentous alpha-synuclein inclusions of alpha-synucleinopathies" 168 : 947-961, 2006

36 Okubo M, "Clonal chromosomal aberrations accompanied by strong telomerase activity in immortalization of human B-lymphoblastoid cell lines transformed by Epstein-Barr virus" 129 : 30-34, 2001

37 Ravid R, "Brain banks as key part of biochemical and molecular studies on cerebral cortex involvement in Parkinson’s disease" 279 : 1167-1176, 2012

38 Luo GR, "Are heat shock proteins therapeutic target for Parkinson’s disease?" 3 : 20-26, 2007

39 Anglade P, "Apoptosis and autophagy in nigral neurons of patients with Parkinson’s disease" 12 : 25-31, 1997

40 Miller RM, "Altered gene expression profiles reveal similarities and differences between Parkinson disease and model systems" 11 : 539-549, 2005

41 Sugden PH, "Activities of citrate synthase, NAD+-linked and NADP+-linked isocitrate dehydrogenases, glutamate dehydrogenase, aspartate aminotransferase and alanine aminotransferase in nervous tissues from vertebrates and invertebrates" 150 : 105-111, 1975

42 Choi KC, "A novel mTOR activating protein protects dopamine neurons against oxidative stress by repressing autophagy related cell death" 112 : 366-376, 2010

43 Cooperstein SJ, "A microspectrophotometric method for the determination of cytochrome oxidase" 189 : 665-670, 1951

44 Strittmatter CF, "A hemochromogen component of liver microsomes" 38 : 19-25, 1952