title: PTEN regulates fibroblast elimination during collagen matrix contraction.
abstract: During tissue repair, excess fibroblasts are eliminated by apoptosis. This physiologic process limits fibrosis and restores normal anatomic patterns. Replicating physiologic apoptosis associated with tissue repair, fibroblasts incorporated into type I collagen matrices undergo apoptosis in response to collagen matrix contraction. In this in vitro model of wound repair, fibroblasts first attach to collagen via alpha2beta1 integrin. This provides a survival signal via activation of the phosphatidylinositol 3-kinase/Akt signal pathway. However, during subsequent collagen matrix contraction, the level of phosphorylated Akt progressively declines, triggering apoptosis. The mechanism underlying the fall in phosphorylated Akt is incompletely understood. Here we show that PTEN phosphatase becomes activated during collagen matrix contraction and is responsible for antagonizing phosphatidylinositol 3-kinase activity and promoting a decline in phosphorylated Akt and fibroblast apoptosis in response to collagen contraction. PTEN null fibroblasts displayed enhanced levels of phosphorylated Akt and were resistant to collagen matrix contraction-induced apoptosis. Reconstitution of PTEN in PTEN null cells conferred susceptibility to apoptosis in response to contraction of collagen matrices. Consistent with this, knockdown of PTEN in PTEN(+/+) embryonic fibroblasts by small interfering RNA augmented Akt activity and suppressed apoptosis in contractile collagen matrices. Furthermore, inhibition of Akt activity restored the sensitivity of PTEN null cells to collagen contraction-induced apoptosis, indicating that the mechanism by which PTEN alters fibroblast viability is through modulation of phosphorylated Akt levels. Our work suggests that collagen matrix contraction activates PTEN by a mechanism involving cytoskeletal disassembly. Our studies indicate a key role for PTEN in regulating fibroblast viability during tissue repair.
authors.affiliation: Department of Medicine, University of Minnesota, Minneapolis, Minnesota 55455, USA. nhoxx002@umn.edu
authors.forename: Richard Seonghun
authors.initials: RS
authors.lastname: Nho
authors.forename: Hong
authors.initials: H
authors.lastname: Xia
authors.forename: Deanna
authors.initials: D
authors.lastname: Diebold
authors.forename: Judy
authors.initials: J
authors.lastname: Kahm
authors.forename: Jill
authors.initials: J
authors.lastname: Kleidon
authors.forename: Eric
authors.initials: E
authors.lastname: White
authors.forename: Craig A
authors.initials: CA
authors.lastname: Henke
mesh_terms: D000199:Actins / Q000378:metabolism; D017209:Apoptosis; D002460:Cell Line; D003094:Collagen / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts; D005786:Gene Expression Regulation; D006801:Humans; D008168:Lung / Q000166:cytology / Q000201:enzymology; D008870:Microtubules / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D051059:PTEN Phosphohydrolase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D000199:Actins; D034741:RNA, Small Interfering; D003094:Collagen; D051057:Proto-Oncogene Proteins c-akt; D051059:PTEN Phosphohydrolase