Role of type II pneumocyte senescence in radiation-induced lung fibrosis.
BACKGROUND
Radiation is a commonly delivered therapeutic modality for cancer. The causes underlying the chronic, progressive nature of radiation injury in the lung are poorly understood.


METHODS
C57Bl/6NCr mice were exposed to thoracic irradiation (n = 3 per dose and time point for tissue collection). Microarray analysis of gene expression from irradiated murine lung was performed using one-way analysis of variance with post hoc Scheffe analysis. Senescence and type II airway epithelial cell (AECII) count were assayed in irradiated murine lung tissue (n = 3 per condition). Irradiated mice were treated with diphenyleneiodonium (DPI), an inhibitor of NADPH oxidase (NOX), and fibrosis was assessed by collagen assays. All statistical tests were two-tailed.


RESULTS
Gene expression in lung tissue from mice irradiated to 17.5 Gy clustered with that of aged unirradiated mice. Only fibrogenic exposures led to AECII senescence (0 Gy: 0.66% ± 0.67%; 5 Gy: 4.5% ± 1.19%; 17.5 Gy: 18.7% ± 3.05; P = .007) and depletion (0 Gy: 2.89 per alveolus ± 0.26; 5 Gy: 2.41 ± 0.19; 17.5 Gy: 1.6 ± 0.14; P < .001) at 30 weeks. Treatment of irradiated mice with DPI for 16 weeks markedly reduced collagen accumulation (5×6 Gy: 57.26 μg/lung ± 9.91; 5×6 Gy ± DPI: 36.54μg/lung ± 4.39; P = .03) and AECII senescence (5×6 Gy: 37.61% ± 4.82%; 5×6 Gy ± DPI: 12.38% ± 2.78; P < .001).


CONCLUSIONS
These studies identify senescence as an important process in AECII in vivo and indicate that NOX is a critical mediator of radiation-induced AECII senescence and pulmonary fibrosis.
Affiliations of authors: Radiation Oncology Branch (DEC, US, JAH, WS, SZ, HA, AY, EJC) and Radiation Biology Branch (AS, AT), Center for Cancer Research, National Institutes of Health, Bethesda, MD.
Deborah E
DE
Citrin
Uma
U
Shankavaram
Jason A
JA
Horton
William
W
Shield
Shuping
S
Zhao
Hiroaki
H
Asano
Ayla
A
White
Anastasia
A
Sowers
Angela
A
Thetford
Eun Joo
EJ
Chung
D056809:Alveolar Epithelial Cells / Q000473:pathology* / Q000528:radiation effects*; D000818:Animals; D016922:Cellular Senescence / Q000528:radiation effects; D003094:Collagen / Q000378:metabolism*; D004195:Disease Models, Animal; D004307:Dose-Response Relationship, Radiation; D005786:Gene Expression Regulation / Q000528:radiation effects; D008168:Lung / Q000473:pathology / Q000528:radiation effects; D051379:Mice; D008810:Mice, Inbred C57BL; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009861:Onium Compounds; D011658:Pulmonary Fibrosis / Q000209:etiology* / Q000473:pathology*; D013997:Time Factors; D046888:Tissue Array Analysis
D009861:Onium Compounds; C007517:diphenyleneiodonium; D003094:Collagen; D019255:NADPH Oxidases