Structural stability of human fibroblast growth factor-1 is essential for protective effects against radiation-induced intestinal damage.
OBJECTIVE
Human fibroblast growth factor-1 (FGF1) has radioprotective effects on the intestine, although its structural instability limits its potential for practical use. Several stable FGF1 mutants were created increasing stability in the order, wild-type FGF1, single mutants (Q40P, S47I, and H93G), Q40P/S47I, and Q40P/S47I/H93G. This study evaluated the contribution of the structural stability of FGF1 to its radioprotective effect.


METHODS
Each FGF1 mutant was administered intraperitoneally to BALB/c mice in the absence of heparin 24 h before or after total body irradiation (TBI) with γ-rays at 8-12 Gy. Several radioprotective effects were examined in the jejunum.


RESULTS
Q40P/S47I/H93G could activate all subtypes of FGF receptors in vitro much more strongly than the wild-type without endogenous or exogenous heparin. Preirradiation treatment with Q40P/S47I/H93G significantly increased crypt survival more than wild-type FGF1 after TBI at 10 or 12 Gy, and postirradiation treatment with Q40P/S47I/H93G was effective in promoting crypt survival after TBI at 10, 11, or 12 Gy. In addition, crypt cell proliferation, crypt depth, and epithelial differentiation were significantly promoted by postirradiation treatment with Q40P/S47I/H93G. The level of stability of FGF1 mutants correlated with their mitogenic activities in vitro in the absence of heparin; however, preirradiation treatment with the mutants increased the crypt number to almost the same level as Q40P/S47I/H93G. When given 24 h after TBI at 10 Gy, all FGF1 mutants increased crypt survival more than wild-type FGF1, and Q40P/S47I/H93G had the strongest mitogenic effects in intestinal epithelial cells after radiation damage. Moreover, Q40P/S47I/H93G prolonged mouse survival after TBI because of the repair of intestinal damage.


CONCLUSIONS
These findings suggest that the structural stability of FGF1 can contribute to the enhancement of protective effects against radiation-induced intestinal damage. Therefore, Q40P/S47I/H93G is pharmacologically one of the most promising candidates for clinical applications for radiation-induced gastrointestinal syndrome.
Advanced Radiation Biology Research Program, Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, Chiba, Japan. f_naka@nirs.go.jp
Fumiaki
F
Nakayama
Sachiko
S
Umeda
Takeshi
T
Yasuda
Masahiro
M
Asada
Kaori
K
Motomura
Masashi
M
Suzuki
Malgorzata
M
Zakrzewska
Toru
T
Imamura
Takashi
T
Imai
D000818:Animals; D016220:Fibroblast Growth Factor 1 / Q000008:administration & dosage / Q000235:genetics*; D006493:Heparin / Q000494:pharmacology; D007422:Intestines / Q000187:drug effects / Q000528:radiation effects*; D007583:Jejunum / Q000187:drug effects / Q000528:radiation effects; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D011833:Radiation Injuries, Experimental / Q000517:prevention & control*; D011837:Radiation-Protective Agents / Q000008:administration & dosage; D017468:Receptors, Fibroblast Growth Factor / Q000187:drug effects / Q000378:metabolism; D014916:Whole-Body Irradiation
D011837:Radiation-Protective Agents; D017468:Receptors, Fibroblast Growth Factor; D016220:Fibroblast Growth Factor 1; D006493:Heparin