Spatial control of Cdc42 signalling by a GM130–RasGRF complex regulates polarity and tumorigenesis

Baschieri, Francesco; Confalonieri, Stefano; Bertalot, Giovanni; Fiore, Pier Paolo Di; Dietmaier, Wolfgang; Leist, Marcel; Crespo, Piero; Macara, Ian G.; Farhan, Hesso

doi:10.1038/ncomms5839

Cited by 84 publications

(125 citation statements)

References 42 publications

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“…Loss of epithelial cell polarity contributes to acquisition of an invasive phenotype in models of human breast cancer 43 ; thus, we evaluated the expression of GM130, a Golgi epithelial cell polarity marker whose expression is frequently lost in breast cancer. 44 IHC analysis of GM130 did not show differences in localization between normal epithelium or DCIS tumor cells regardless of myoepithelial cell marker expression, nor were differences in E-cadherin staining observed (data not shown). However, IHC staining of calponin and p63 suggested an inverse relation between myoepithelial cell expression of calponin and gain of p63 expression in adjacent tumor cells (Figure 6, AeD).…”

Section: Characterization Of Myoepithelial Cells With Dcis Progressiomentioning

confidence: 88%

Myoepithelial Cell Differentiation Markers in Ductal Carcinoma in Situ Progression

Russell

Jindal

Agunbiade

et al. 2015

The American Journal of Pathology

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We describe a preclinical model that investigates progression of early-stage ductal carcinoma in situ (DCIS) and report that compromised myoepithelial cell differentiation occurs before transition to invasive disease. Human breast cancer MCF10DCIS.com cells were delivered into the mouse mammary teat by intraductal injection in the absence of surgical manipulations and accompanying wound-healing confounders. DCIS-like lesions developed throughout the mammary ducts with full representation of human DCIS histologic patterns. Tumor cells were incorporated into the normal mammary epithelium, developed ductal intraepithelial neoplasia and DCIS, and progressed to invasive carcinoma, suggesting the model provides a rigorous approach to study early stages of breast cancer progression. Mammary glands were evaluated for myoepithelium integrity with immunohistochemical assays. Progressive loss of the myoepithelial cell differentiation markers p63, calponin, and a-smooth muscle actin was observed in the mouse myoepithelium surrounding DCISinvolved ducts. p63 loss was an early indicator, calponin loss intermediate, and a-smooth muscle actin a later indicator of compromised myoepithelium. Loss of myoepithelial calponin was specifically associated with gain of the basal marker p63 in adjacent tumor cells. In single time point biopsies obtained from 16 women diagnosed with pure DCIS, a similar loss in myoepithelial cell markers was observed. These results suggest that further research is warranted into the role of myoepithelial cell p63 and calponin expression on DCIS progression to invasive disease. Clinical evidence is compelling for histologic progression of breast cancer through atypical hyperplasia, ductal carcinoma in situ (DCIS), invasive ductal carcinoma, and metastatic stages. 1 Such histopathologic progression studies and mutational profiling of epithelial cancers 2,3 suggest that acquisition of invasive potential is a relatively late event. However, genomic data analyses have revealed that most tumor cell gene expression changes occur at the transition from normal to DCIS, with few additional changes in expression occurring at the transition from DCIS to overt invasive disease. 4,5 These observations implicate key roles for nonepithelial cells in progression to invasive disease. 6,7 The lack of relevant model systems has hindered our understanding of the DCIS to invasive transition.The clinical definition of invasive breast cancer is spread of malignant tumor cells from the confines of the mammary duct into the adjacent tissue stroma. In the normal mammary gland, epithelial ductal and alveolar structures are

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Section: Characterization Of Myoepithelial Cells With Dcis Progressiomentioning

confidence: 88%

Myoepithelial Cell Differentiation Markers in Ductal Carcinoma in Situ Progression

Russell

Jindal

Agunbiade

et al. 2015

The American Journal of Pathology

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“…Cell-culture-based studies have implicated GM130 in a number of cellular processes in addition to secretory trafficking, including cytoskeletal regulation, which is important for cell migration and cell division (4,31,33,34). It was therefore surprising that the GM130 KO mice did not display any overt developmental phenotype; pups were born at normal weight and looked morphologically normal.…”

Section: Discussionmentioning

confidence: 99%

“…GM130 also participates in vesicle tethering during ER-to-Golgi traffic (24,26,27), and can function as a scaffold for activation of Cdc42 or Stk25 that is relevant for cell migration (32)(33)(34). To elucidate the cellular basis of the ataxic phenotype and Purkinje cell degeneration of GM130-nKO mice, we used antibodies to golgin-84 and TGN38, markers of the cis and trans-Golgi respectively, to analyze Golgi structure in Purkinje cells from mice at ages of P8, P14 and P28 by immunostaining.…”

Section: Progressive Cerebellar Atrophy and Purkinje Cell Loss In Gm1mentioning

confidence: 99%

Loss of the golgin GM130 causes Golgi disruption, Purkinje neuron loss, and ataxia in mice

Liu

Mei

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

104

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The Golgi apparatus lies at the heart of the secretory pathway where it is required for secretory trafficking and cargo modification. Disruption of Golgi architecture and function has been widely observed in neurodegenerative disease, but whether Golgi dysfunction is causal with regard to the neurodegenerative process, or is simply a manifestation of neuronal death, remains unclear. Here we report that targeted loss of the golgin GM130 leads to a profound neurological phenotype in mice. Global KO of mouse GM130 results in developmental delay, severe ataxia, and postnatal death. We further show that selective deletion of GM130 in neurons causes fragmentation and defective positioning of the Golgi apparatus, impaired secretory trafficking, and dendritic atrophy in Purkinje cells. These cellular defects manifest as reduced cerebellar size and Purkinje cell number, leading to ataxia. Purkinje cell loss and ataxia first appear during postnatal development but progressively worsen with age. Our data therefore indicate that targeted disruption of the mammalian Golgi apparatus and secretory traffic results in neuronal degeneration in vivo, supporting the view that Golgi dysfunction can play a causative role in neurodegeneration.GM130 | Golgi apparatus | polarized secretion | Purkinje cell | ataxia

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“…Strikingly, consistent with these observations, our immunolabeling assays also uncovered significant accumulation of activated, phosphorylated pPAK, pMLC2 and pVASP in the bodies of cone photoreceptor cells and around the ectopic nuclei protruding into the PS region of GRF2-depleted retinas. Because Rac and CDC42 are known targets for positive or negative regulation by the GRF proteins (Calvo et al, 2011;Feig, 2011;Fernández-Medarde and Santos, 2011) and they have also been shown to exert stage-specific functions in the control of polarity in physiological neurogenesis and in tumorigenesis (Baschieri et al, 2014;Vadodaria et al, 2013), our observations support the notion that the absence of productive functional interactions between GRF2 and its potential downstream targets Rac and CDC42 in GRF2-depleted retinas might lead to subsequent alterations of the formation or structure of PAR-containing, polarity-related macromolecular complexes as well as abnormal activation of related signaling molecules such as PAK (Zhang et al, 1995), VASP (Rich et al, 1997) and MLC2 (Bellion, 2005;Schenk et al, 2009) that could be responsible for the broken OLM and defective cone nuclear migration and final location observed in the GRF2-KO and GRF1/2-DKO retinas.…”

Section: Discussionmentioning

confidence: 99%

RasGRF2 controls nuclear migration in postnatal retinal cone photoreceptors

Jimeno

Gómez

Calzada

et al. 2016

Journal of Cell Science

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Detailed immunocytochemical analyses comparing wild-type (WT), GRF1-knockout (KO), GRF2-KO and GRF1/2 double-knockout (DKO) mouse retinas uncovered the specific accumulation of misplaced, 'ectopic' cone photoreceptor nuclei in the photoreceptor segment (PS) area of retinas from GRF2-KO and GRF1/2-DKO, but not of WT or GRF1-KO mice. Localization of ectopic nuclei in the PS area of GRF2-depleted retinas occurred postnatally and peaked between postnatal day (P)11 and P15. Mechanistically, the generation of this phenotype involved disruption of the outer limiting membrane and intrusion into the PS layer by cone nuclei displaying significant perinuclear accumulation of signaling molecules known to participate in nuclear migration and cytoskeletal reorganization, such as PAR3, PAR6 and activated, phosphorylated forms of PAK, MLC2 and VASP. Electroretinographic recordings showed specific impairment of cone-mediated retinal function in GRF2-KO and GRF1/2-DKO retinas compared with WT controls. These data identify defective cone nuclear migration as a novel phenotype in mouse retinas lacking GRF2 and support a crucial role of GRF2 in control of the nuclear migration processes required for proper postnatal development and function of retinal cone photoreceptors.

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Spatial control of Cdc42 signalling by a GM130–RasGRF complex regulates polarity and tumorigenesis

Cited by 84 publications

References 42 publications

Myoepithelial Cell Differentiation Markers in Ductal Carcinoma in Situ Progression

Myoepithelial Cell Differentiation Markers in Ductal Carcinoma in Situ Progression

Loss of the golgin GM130 causes Golgi disruption, Purkinje neuron loss, and ataxia in mice

RasGRF2 controls nuclear migration in postnatal retinal cone photoreceptors

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