Gene targeting of the cysteine peptidase cathepsin H impairs lung surfactant in mice.
BACKGROUND
The 11 human cysteine cathepsins are proteases mainly located in the endolysosomal compartment of all cells and within the exocytosis pathways of some secretory cell types. Cathepsin H (Ctsh) has amino- and endopeptidase activities. In vitro studies have demonstrated Ctsh involvement in the processing and secretion of the pulmonary surfactant protein B (SP-B). Furthermore, Ctsh is highly expressed in the secretory organelles of alveolar type II pneumocytes where the surfactant proteins are processed.


RESULTS
Hence, we generated Ctsh null mice by gene targeting in embryonic stem cells to investigate the role of this protease in surfactant processing in vivo. The targeting construct contains a ß-galactosidase (lacZ) reporter enabling the visualisation of Ctsh expression sites. Ctsh-deficiency was verified by northern blot, western blot, and measurement of the Ctsh aminopeptidase activity. Ctsh(-/-) mice show no gross phenotype and their development is normal without growth retardation. Broncho-alveolar lavage (BAL) from Ctsh(-/-) mice contained lower levels of SP-B indicating reduced SP-B secretion. The BAL phospholipid concentration was not different in Ctsh(+/+) and Ctsh(-/-) mice, but measurement of surface tension by pulsating bubble surfactometry revealed an impairment of the tension reducing function of lung surfactant of Ctsh(-/-) mice.


CONCLUSIONS
We conclude that cathepsin H is involved in the SP-B production and reduced SP-B levels impair the physical properties of the lung surfactant. However, Ctsh defiency does not reproduce the severe phenotype of SP-B deficient mice. Hence, other proteases of the secretory pathway of type II pneumocytes, i.e. cathepsins C or E, are still able to produce surfactant of sufficient quality in absence of Ctsh.
Institute of Laboratory Medicine, Carl-Thiem-Klinikum, Cottbus, Germany.
Frank
F
Bühling
Martin
M
Kouadio
Caroline E
CE
Chwieralski
Ursula
U
Kern
Jens M
JM
Hohlfeld
Nicole
N
Klemm
Nicole
N
Friedrichs
Wera
W
Roth
Jan M
JM
Deussing
Christoph
C
Peters
Thomas
T
Reinheckel
D000818:Animals; D056655:Cathepsin H / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D018390:Gene Targeting; D006801:Humans; D008168:Lung / Q000201:enzymology / Q000473:pathology; D051379:Mice; D010641:Phenotype; D037661:Pulmonary Surfactant-Associated Proteins / Q000378:metabolism; D011663:Pulmonary Surfactants / Q000378:metabolism*
D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D056655:Cathepsin H