Characterization of cellular senescence mechanisms in human corneal endothelial cells

Aging Cell

Angela N. Sheerin⁴  S. Kaye Smith¹  Katrin Jennert-Burston⁴  Amy J. Brook¹  Marcus C. Allen⁴  Badr Ibrahim⁴  Dawn Jones⁴  Corrin Wallis⁴  Katrin Engelmann²  William Rhys-Williams³  Richard G. A. Faragher⁴ 

[1] School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK;Department of Ophthalmology, Klinikum Chemnitz GmbH, Klinik für Augenheilkunde, Flemmingstraße 2, 09116 Chemnitz, Dresden, Germany;Destiny Pharma Ltd., Sussex Innovation Centre, Science Park Square, Falmer, Brighton BN1 9SB, UK;School of Pharmacy and Biomolecular Sciences, University of Brighton, Huxley Building, Lewes Road, Brighton BN2 4GJ, UK

关键词: senescence;    telomeres;    telomerase;    p53;    oxidative stress;    CDK4;    replicative senescence;   

DOI  :  10.1111/j.1474-9726.2011.00776.x

来源: Wiley

【 摘 要 】

Summary

The human cornea is a tri-laminar structure composed of several cell types with substantial mitotic potential. Age-related changes in the cornea are associated with declining visual acuity and the onset of overt age-related corneal diseases. Corneal transplantation is commonly used to restore vision in patients with damaged or diseased corneas. However, the supply of donor tissue is limited, and thus there is considerable interest in the development of tissue-engineered alternatives. A major obstacle to these approaches is the short replicative lifespan of primary human corneal endothelial cells (HCEC). Accordingly, a comprehensive investigation of the signalling pathways and mechanisms underpinning proliferative lifespan and senescence in HCEC was undertaken. The effects of exogenous human telomerase reverse transcriptase expression, p53 knockdown, disruption of the pRb pathway by over-expression of CDK4 and reduced oxygen concentration on the lifespan of primary HCEC were evaluated. We provide proof-of-principle that forced expression of telomerase, when combined with either p53 knockdown or CDK4 over-expression, is sufficient to produce immortalized HCEC lines. The resultant cell lines express an HCEC-specific transcriptional fingerprint, and retain expression of the corneal endothelial temperature-sensitive potassium channel, suggesting that significant dedifferentiation does not occur as a result of these modes of immortalization. Exploiting these insights into proliferative lifespan barriers in HCEC will underpin the development of novel strategies for cell-based therapies in the human cornea.

【 授权许可】

Unknown   
© 2011 The Authors. Aging Cell © 2011 Blackwell Publishing Ltd/Anatomical Society of Great Britain and Ireland