Loss of integrin linked kinase from mouse hepatocytes in vitro and in vivo results in apoptosis and hepatitis†

Gkretsi, Vasiliki; Mars, Wendy M.; Bowen, William C.; Barua, Lindsay; Yang, Yu Hsuan Carol; Guo, Lei; Arnaud, René; Dedhar, Shoukat; Wu, Chuanyue; Michalopoulos, George K.

doi:10.1002/hep.21540

Cited by 56 publications

(63 citation statements)

References 48 publications

(75 reference statements)

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“…We generated several ILK-knockout (ILK 2/2 ) clonal lines by infecting immortalized newborn ILK fl/fl mouse fibroblasts with adeno-Cre recombinase. Immunoblotting confirmed ILK knockout and, as previously reported (Fukuda et al, 2003;Gkretsi et al, 2007;Wang et al, 2008), this also led to loss of the ILK-interacting proteins PINCH1 and a-parvin (Fig. 7B).…”

Section: Ilk Binding Aids Kindlin-2 Fa Targetingsupporting

confidence: 87%

“…ILK partners with the LIM domain protein PINCH (through its ARD) and the calponin homology (CH) domain protein parvin (through its kinase domain) (Chiswell et al, 2010;Chiswell et al, 2008;Fukuda et al, 2009;Stiegler et al, 2013) and this triprotein IPP complex is central to ILK function (Fukuda et al, 2003;Gkretsi et al, 2007;Stanchi et al, 2009;Wang et al, 2008). In addition to PINCH and parvin, ILK interacts directly or indirectly with a range of other FA proteins and is required to maintain FA architecture and undertake integrin-mediated signaling (Elad et al, 2013;Kogata et al, 2009;Sakai et al, 2003;Wickström et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Differential binding to the ILK complex determines kindlin isoform adhesion localization and integrin activation

Huet-Calderwood

Brahme

Kumar

et al. 2014

Journal of Cell Science

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BSTRACTKindlins are essential FERM-domain-containing focal adhesion (FA) proteins required for proper integrin activation and signaling. Despite the widely accepted importance of each of the three mammalian kindlins in cell adhesion, the molecular basis for their function has yet to be fully elucidated, and the functional differences between isoforms have generally not been examined. Here, we report functional differences between kindlin-2 and -3 (also known as FERMT2 and FERMT3, respectively); GFP-tagged kindlin-2 localizes to FAs whereas kindlin-3 does not, and kindlin-2, but not kindlin-3, can rescue a5b1 integrin activation defects in kindlin-2-knockdown fibroblasts. Using chimeric kindlins, we show that the relatively uncharacterized kindlin-2 F2 subdomain drives FA targeting and integrin activation. We find that the integrin-linked kinase (ILK)-PINCH-parvin complex binds strongly to the kindlin-2 F2 subdomain but poorly to that of kindlin-3. Using a point-mutated kindlin-2, we establish that efficient kindlin-2-mediated integrin activation and FA targeting require binding to the ILK complex. Thus, ILK-complex binding is crucial for normal kindlin-2 function and differential ILK binding contributes to kindlin isoform specificity.

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Section: Ilk Binding Aids Kindlin-2 Fa Targetingsupporting

confidence: 87%

Section: Introductionmentioning

confidence: 99%

Differential binding to the ILK complex determines kindlin isoform adhesion localization and integrin activation

Huet-Calderwood

Brahme

Kumar

et al. 2014

Journal of Cell Science

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“…Furthermore, the interaction of cells with the components of the Matrigel might play important roles in the recovery partly through the modulation of expression of transcription factors, including HNF-4α, 19 and the activation of integrin-linked kinase that is involved in transmitting integrin signaling to the interior of the cell. 20,21 The recovery of albumin expression in transdifferentiated hepatocytes was enhanced by Dex, which has been shown to increase albumin and TAT expression in spheroidal aggregate cultures of newborn rat hepatocytes 18 and decrease CK 19 mRNA expression in monolayer-cultured rat hepatocytes. 7 Although the mechanism of the effect of Dex on hepatocytic phenotypes has not been elucidated, this study suggests that the inhibition of the MEK and JNK signaling pathways may be important.…”

Section: Discussionmentioning

confidence: 99%

Recovery of Mature Hepatocytic Phenotype following Bile Ductular Transdifferentiation of Rat Hepatocytes in Vitro

Sone

Nishikawa

Nagahama

et al. 2012

The American Journal of Pathology

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We previously demonstrated that mature rat hepatocytes transdifferentiate to bile ductular cells when cultured in a three-dimensional collagen-rich matrix. Here, we show that the phenotype of transdifferentiated hepatocytes can be reversed by modulating culture conditions. Spheroidal aggregates of hepatocytes were cultured within a collagen gel matrix in the presence of serum and tumor necrosis factor-α. Spheroids transformed into ductular structures composed of small cuboidal cells, lost the expression of hepatocytic markers, while aberrantly expressed bile ductular markers. The transdifferentiated cells were then retrieved from the gels, plated on Matrigel-coated surfaces, and cultured in serum-free media. Cells spontaneously formed spheroidal aggregates and recovered hepatocytic phenotype. While Dexamethasone (Dex), which suppressed the phosphorylation of ERK and Jun N-terminal kinase, facilitated the recovery, the combination with interleukin-6 or oncostatin M resulted in the recovery of HNF-4α protein expression and the typical hepatocytic morphology and a decrease in the expression of bile ductular markers. A cDNA microarray analysis revealed that the hepatocyte-specific mRNA expression profile was recovered in these cells. Our results demonstrate that hepatocytes are able to recover their phenotypes following bile ductular transdifferentiation, suggesting that hepatocytic and bile ductular phenotypes may be mutually reversible.4

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“…Moreover, several reports demonstrate that ILK has a role in mitosis by regulation of the microtubule cytoskeleton. Mitotic defects have been detected upon ILK depletion in Drosophila S2 cells (Bettencourt-Dias et al, 2004), mouse hepatocytes (Gkretsi et al, 2007) and human glioblastoma cells (Koul et al, 2005;Edwards et al, 2008). It is interesting that, although the cellular phenotypes were not reported in detail in these studies, all of these systems contain supernumerary centrosomes (Weber et al, 1998;Guidotti et al, 2003;Margall-Ducos et al, 2007;Kwon et al, 2008).…”

Section: Introductionmentioning

confidence: 88%

A critical role of integrin-linked kinase, ch-TOG and TACC3 in centrosome clustering in cancer cells

et al. 2010

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Many cancer cells contain more than two centrosomes, which imposes a potential for multipolar mitoses, leading to cell death. To circumvent this, cancer cells develop mechanisms to cluster supernumerary centrosomes to form bipolar spindles, enabling successful mitosis. Disruption of centrosome clustering thus provides a selective means of killing supernumerary centrosome-harboring cancer cells. Although the mechanisms of centrosome clustering are poorly understood, recent genetic analyses have identified requirements for both actin and tubulin regulating proteins. In this study, we demonstrate that the integrin-linked kinase (ILK), a protein critically involved in actin and mitotic microtubule organization, is required for centrosome clustering. Inhibition of ILK expression or activity inhibits centrosome clustering in several breast and prostate cancer cell lines that have centrosome amplification. Furthermore, cancer cells with supernumerary centrosomes are significantly more sensitive to ILK inhibition than cells with two centrosomes, demonstrating that inhibiting ILK offers a selective means of targeting cancer cells. Live cell analysis shows ILK perturbation leads cancer cells to undergo multipolar anaphases, mitotic arrest and cell death in mitosis. We also show that ILK performs its centrosome clustering activity in a focal adhesion-independent, but centrosome-dependent, manner through the microtubule regulating proteins TACC3 and ch-TOG. In addition, we identify a specific TACC3 phosphorylation site that is required for centrosome clustering and demonstrate that ILK regulates this phosphorylation in an Aurora-A-dependent manner.

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Loss of integrin linked kinase from mouse hepatocytes in vitro and in vivo results in apoptosis and hepatitis†

Cited by 56 publications

References 48 publications

Differential binding to the ILK complex determines kindlin isoform adhesion localization and integrin activation

Differential binding to the ILK complex determines kindlin isoform adhesion localization and integrin activation

Recovery of Mature Hepatocytic Phenotype following Bile Ductular Transdifferentiation of Rat Hepatocytes in Vitro

A critical role of integrin-linked kinase, ch-TOG and TACC3 in centrosome clustering in cancer cells

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