Age-related transcription levels of KU70, MGST1 and BIK in CD34+ hematopoietic stem and progenitor cells.
Despite the known longevity of human hematopoietic stem and progenitor cells (HSC), numerous functional impairments of these cells can be observed in an age-dependent manner. However, the molecular alterations associated with aging of HSC are largely unknown. Therefore, we scrutinized gene expression patterns of HSC from newborn, young and old healthy donors. CD34+ HSC were isolated via immuno-magnetic separation and evaluated by FACS analysis. We performed cDNA macroarray analyses on a first set of CD34+ samples (n=13). We found the genes encoding KU-antigen 70 kD (KU70), microsomal glutathione S-transferase 1 (MGST1) and BCL2-interacting killer (BIK) to possess age-related mRNA expression levels. KU70 is a DNA repair gene and part of the DNA-dependent protein kinase (DNA-PK) complex. Its expression was negatively correlated with donor age showing highest expression levels in newborn, 2.6-fold lower levels in young and 6.3-fold lower levels in old donors. The transcription levels of MGST1, a gene protecting against oxidative stress, progressively increased with age. Expression was lowest in newborn, 2.6-fold higher in young and 4.3-fold higher in old donors. BIK is a proapoptotic gene and its expression was positively correlated with donor age: lowest in newborn, 1.8-fold higher in young and 4.1-fold higher in old donors. These findings were confirmed with an independent, second set of CD34+ samples (n=16) by means of quantitative real-time RT-PCR. Elucidation of age-dependent molecular alterations in healthy HSC facilitate a better understanding of functional impairments in hematopoiesis and may become valuable for anti-aging drug development and the emerging field of regenerative medicine.
Prall
Wolf C
WC
Department of Hematology, Oncology and Clinical Immunology, Heinrich-Heine-University of Duesseldorf, 40225 Duesseldorf, Germany. wprall@BIDMC.harvard.edu
Czibere
Akos
A
Jäger
Marcus
M
Spentzos
Dimitrios
D
Libermann
Towia A
TA
Gattermann
Norbert
N
Haas
Rainer
R
Aivado
Manuel
M
D000328:Adult; D053687:Adult Stem Cells / Q000737:chemistry*; D000367:Age Factors; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000235:genetics*; D018952:Antigens, CD34 / Q000032:analysis*; D034961:Antigens, Nuclear / Q000032:analysis* / Q000235:genetics; D051017:Apoptosis Regulatory Proteins / Q000032:analysis* / Q000235:genetics; D016922:Cellular Senescence / Q000235:genetics; D016000:Cluster Analysis; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000235:genetics; D005312:Fetal Blood / Q000166:cytology; D020869:Gene Expression Profiling / Q000379:methods; D005982:Glutathione Transferase / Q000032:analysis* / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000737:chemistry*; D006801:Humans; D007231:Infant, Newborn; D000072200:Ku Autoantigen; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics; D008875:Middle Aged; D024101:Mitochondrial Proteins; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D015203:Reproducibility of Results; D014158:Transcription, Genetic
D018952:Antigens, CD34; D034961:Antigens, Nuclear; D051017:Apoptosis Regulatory Proteins; C096820:BIK protein, human; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; C122107:microsomal glutathione S-transferase-I; D005982:Glutathione Transferase; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen