Document Detail


Nucleus pulposus cellular longevity by telomerase gene therapy.
MedLine Citation:
PMID:  17495775     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
STUDY DESIGN: Nonviral transfection of nucleus pulposus cells with a telomerase expression construct to assess the effects on cellular lifespan, function, karyotypic stability, and transformation properties. OBJECTIVES: To investigate whether telomerase gene therapy can extend the cellular lifespan while retaining functionality of nucleus pulposus cells in a safe manner. SUMMARY OF BACKGROUND DATA: Degeneration of the intervertebral disc is an age-related condition in which cells responsible for the maintenance and health of the disc deteriorate with age. Telomerase can extend the cellular lifespan and function of other musculoskeletal tissues, such as the heart, bones, and connective tissues. Therefore, extension of the cellular lifespan and matrix production of intervertebral disc cells may have the potential to delay the degeneration process. METHODS: Ovine nucleus pulposus cells were lipofectamine transfected in vitro with a human telomerase reverse transcriptase (hTERT) expression construct. Cellular lifespan and matrix transcript levels were determined by cumulative population doublings and real-time RT-PCR, respectively. G1-cell cycle checkpoint, p53 functionality, growth of transfected cells in anchorage-independent or serum starvation conditions, and karyotypic analysis were performed. RESULTS: Transfection was achieved successfully with 340% +/- 7% (mean +/- SD) relative telomerase activity in hTERT-transfected cells. hTERT transfection enabled a 50% extension in mean cellular lifespan and prolonged matrix production of collagen 1 and 2 for more than 282 days. Karyotypic instability was detected but G1-cell cycle checkpoint and p53 was functionally comparable to parental cells with no growth in serum starvation or anchorage-independent conditions. CONCLUSIONS: Telomerase can extend the cellular lifespan of nucleus pulposus cells and prolong the production of extracellular matrix. Safety is still unresolved, as karyotypic instability was detected but no loss of contact inhibition, mitogen dependency, or G1-cell cycle checkpoint control was evident.
Authors:
Sylvia A Chung; Ai Qun Wei; David E Connor; Graham C Webb; Timothy Molloy; Marina Pajic; Ashish D Diwan
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Spine     Volume:  32     ISSN:  1528-1159     ISO Abbreviation:  Spine     Publication Date:  2007 May 
Date Detail:
Created Date:  2007-05-14     Completed Date:  2007-06-04     Revised Date:  2009-07-09    
Medline Journal Info:
Nlm Unique ID:  7610646     Medline TA:  Spine (Phila Pa 1976)     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1188-96     Citation Subset:  IM    
Affiliation:
Orthopaedic Research Institute, University of New South Wales, St. George Hospital Campus, NSW, Australia.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Aging* / genetics
Cell Proliferation
Cell Transformation, Neoplastic / genetics,  metabolism
Cells, Cultured
Chromosome Aberrations
Collagen / genetics,  metabolism
Gene Expression
Gene Therapy / methods*
Humans
Intervertebral Disk / cytology,  metabolism*
Lipids
RNA, Messenger / metabolism
Sheep
Spinal Diseases / genetics,  metabolism,  therapy*
Telomerase / genetics,  metabolism*
Time Factors
Transfection / methods*
Chemical
Reg. No./Substance:
0/Lipids; 0/Lipofectamine; 0/RNA, Messenger; 9007-34-5/Collagen; EC 2.7.7.49/TERT protein, human; EC 2.7.7.49/Telomerase

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