Inhibition of Pkhd1 impairs tubulomorphogenesis of cultured IMCD cells.
Fibrocystin/polyductin (FPC), the gene product of PKHD1, is responsible for autosomal recessive polycystic kidney disease (ARPKD). This disease is characterized by symmetrically large kidneys with ectasia of collecting ducts. In the kidney, FPC predominantly localizes to the apical domain of tubule cells, where it associates with the basal bodies/primary cilia; however, the functional role of this protein is still unknown. In this study, we established stable IMCD (mouse inner medullary collecting duct) cell lines, in which FPC was silenced by short hairpin RNA inhibition (shRNA). We showed that inhibition of FPC disrupted tubulomorphogenesis of IMCD cells grown in three-dimensional cultures. Pkhd1-silenced cells developed abnormalities in cell-cell contact, actin cytoskeleton organization, cell-ECM interactions, cell proliferation, and apoptosis, which may be mediated by dysregulation of extracellular-regulated kinase (ERK) and focal adhesion kinase (FAK) signaling. These alterations in cell function in vitro may explain the characteristics of ARPKD phenotypes in vivo.
Department of Medicine, Vanderbilt University, Nashville, TN 37232, USA.
Weiyi
W
Mai
Dong
D
Chen
Tianbing
T
Ding
Ingyu
I
Kim
Sujun
S
Park
Sae-youll
SY
Cho
Julia S F
JS
Chu
Dan
D
Liang
Ning
N
Wang
Dianqing
D
Wu
Song
S
Li
Ping
P
Zhao
Roy
R
Zent
Guanqing
G
Wu
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology; D002450:Cell Communication / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D002923:Cilia / Q000502:physiology; D004285:Dogs; D048049:Extracellular Signal-Regulated MAP Kinases / Q000502:physiology; D051418:Focal Adhesion Kinase 2 / Q000502:physiology; D016023:Integrins / Q000502:physiology; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000201:enzymology / Q000473:pathology*; D051379:Mice; D017044:Polycystic Kidney, Autosomal Recessive / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D034622:RNA Interference; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology
D016023:Integrins; C484365:Pkhd1 protein, mouse; D011956:Receptors, Cell Surface; D051418:Focal Adhesion Kinase 2; D048049:Extracellular Signal-Regulated MAP Kinases