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Modelling TFE renal cell carcinoma in mice reveals a critical role of WNT signaling

View Article: PubMed Central - PubMed

ABSTRACT

TFE-fusion renal cell carcinomas (TFE-fusion RCCs) are caused by chromosomal translocations that lead to overexpression of the TFEB and TFE3 genes (Kauffman et al., 2014). The mechanisms leading to kidney tumor development remain uncharacterized and effective therapies are yet to be identified. Hence, the need to model these diseases in an experimental animal system (Kauffman et al., 2014). Here, we show that kidney-specific TFEB overexpression in transgenic mice, resulted in renal clear cells, multi-layered basement membranes, severe cystic pathology, and ultimately papillary carcinomas with hepatic metastases. These features closely recapitulate those observed in both TFEB- and TFE3-mediated human kidney tumors. Analysis of kidney samples revealed transcriptional induction and enhanced signaling of the WNT β-catenin pathway. WNT signaling inhibitors normalized the proliferation rate of primary kidney cells and significantly rescued the disease phenotype in vivo. These data shed new light on the mechanisms underlying TFE-fusion RCCs and suggest a possible therapeutic strategy based on the inhibition of the WNT pathway.

Doi:: http://dx.doi.org/10.7554/eLife.17047.001

No MeSH data available.


Related in: MedlinePlus

Characterization of cyst origin in Cdh16Cre::Tfebfs and Cdh16CreErt2::Tfebfs mice.IHC staining of megalin, THP and AQP2 at different time points. Insets are enlargements of representative areas of interest. Larger cysts (denoted by an asterisk) are negative for all the markers tested. DTcy = Distal Tubules cysts; CDcy = Collecting Ducts cysts.DOI:http://dx.doi.org/10.7554/eLife.17047.005
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fig1s3: Characterization of cyst origin in Cdh16Cre::Tfebfs and Cdh16CreErt2::Tfebfs mice.IHC staining of megalin, THP and AQP2 at different time points. Insets are enlargements of representative areas of interest. Larger cysts (denoted by an asterisk) are negative for all the markers tested. DTcy = Distal Tubules cysts; CDcy = Collecting Ducts cysts.DOI:http://dx.doi.org/10.7554/eLife.17047.005

Mentions: At sacrifice, kidneys from adult Cdh16Cre::Tfebfs and tamoxifen-treated Cdh16CreErt2::Tfebfs mice completely filled the abdominal cavity (Figure 1A). An increase in kidney size from Cdh16Cre::Tfebfs mice was observed starting at P12, with a sensible increase in size detected at P30 (Figure 1B). A striking increase in the Kidney to Body Weight (KW/BW) ratio was also observed at this stage (Figure 1C). A severe enlargement of the kidneys and a significant increase in the Kidney to Body Weight (KW/BW) ratio were also observed in Cdh16CreErt2::Tfebfs mice induced with tamoxifen at several developmental stages (P12, P14, P30) (Figure 1—figure supplement 2A and B). These abnormalities were less severe in mice induced at P30 (Figure 1—figure supplement 2B). Survival time of Cdh16Cre::Tfebfs mice was approximately 3 months (Figure 1D). Interestingly, a late induction of Tfeb overexpression in Cdh16CreErt2::Tfebfs mice resulted in a slower development of the phenotype, with less severe kidney enlargement and overall increase in the survival rate (Figure 1D). Renal function from Cdh16Cre::Tfebfs and Cdh16CreErt2::Tfebfs mice was severely affected, as observed by the strong increase in blood urea and albuminuria (Figure 1—figure supplement 2C). High-frequency ultrasound and histological analysis of kidneys from both Cdh16Cre::Tfebfs and Cdh16CreErt2::Tfebfs mice revealed the presence of a severe cystic disease (Figure 1E, Figure 1—figure supplement 2D and E). In Cdh16Cre::Tfebfs mice, small cysts arose mainly from the cortex and outer medulla at P12 and became significantly enlarged at P30. At P90, kidney architecture was completely disrupted by cysts (Figure 1F). Cdh16CreErt2::Tfebfs mice induced at P12 with tamoxifen and sacrificed at P90 showed a higher number of smaller cysts in both cortex and outer medulla (Figure 1F). Cysts were also observed in Cdh16CreErt2::Tfebfs induced at P14 and, to a lesser extent, at P30 (Figure 1—figure supplement 2E). Tubular epithelial cells lining the cysts showed high levels of cadherin 16, indicating the presence of Cdh16Cre-mediated Tfeb overexpression in these cells (Figure 1G). Histological analysis revealed that cysts from Cdh16Cre::Tfebfs mice were positive for AQP2 and THP and negative for megalin, indicating that they originate from collecting ducts and distal tubules and not from proximal tubules. Notably, the largest cysts were almost completely negative to all tubular markers, suggesting that they became undifferentiated. Conversely, cysts from Cdh16CreErt2::Tfebfs mice were positive to megalin and THP, indicating that they arose from proximal and distal tubules (Figure 1H, Figure 1—figure supplement 3). These differences in cyst origin have already been described in other polycystic kidney disease mouse models and have been attributed to intrinsic differences of specific renal segments at different developmental stages (Lantinga-van Leeuwen et al., 2007; Happé et al., 2009; Leonhard et al., 2016; Piontek et al., 2007).10.7554/eLife.17047.002Figure 1.Tfeb overexpressing mice display cystic kidneys.


Modelling TFE renal cell carcinoma in mice reveals a critical role of WNT signaling
Characterization of cyst origin in Cdh16Cre::Tfebfs and Cdh16CreErt2::Tfebfs mice.IHC staining of megalin, THP and AQP2 at different time points. Insets are enlargements of representative areas of interest. Larger cysts (denoted by an asterisk) are negative for all the markers tested. DTcy = Distal Tubules cysts; CDcy = Collecting Ducts cysts.DOI:http://dx.doi.org/10.7554/eLife.17047.005
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fig1s3: Characterization of cyst origin in Cdh16Cre::Tfebfs and Cdh16CreErt2::Tfebfs mice.IHC staining of megalin, THP and AQP2 at different time points. Insets are enlargements of representative areas of interest. Larger cysts (denoted by an asterisk) are negative for all the markers tested. DTcy = Distal Tubules cysts; CDcy = Collecting Ducts cysts.DOI:http://dx.doi.org/10.7554/eLife.17047.005
Mentions: At sacrifice, kidneys from adult Cdh16Cre::Tfebfs and tamoxifen-treated Cdh16CreErt2::Tfebfs mice completely filled the abdominal cavity (Figure 1A). An increase in kidney size from Cdh16Cre::Tfebfs mice was observed starting at P12, with a sensible increase in size detected at P30 (Figure 1B). A striking increase in the Kidney to Body Weight (KW/BW) ratio was also observed at this stage (Figure 1C). A severe enlargement of the kidneys and a significant increase in the Kidney to Body Weight (KW/BW) ratio were also observed in Cdh16CreErt2::Tfebfs mice induced with tamoxifen at several developmental stages (P12, P14, P30) (Figure 1—figure supplement 2A and B). These abnormalities were less severe in mice induced at P30 (Figure 1—figure supplement 2B). Survival time of Cdh16Cre::Tfebfs mice was approximately 3 months (Figure 1D). Interestingly, a late induction of Tfeb overexpression in Cdh16CreErt2::Tfebfs mice resulted in a slower development of the phenotype, with less severe kidney enlargement and overall increase in the survival rate (Figure 1D). Renal function from Cdh16Cre::Tfebfs and Cdh16CreErt2::Tfebfs mice was severely affected, as observed by the strong increase in blood urea and albuminuria (Figure 1—figure supplement 2C). High-frequency ultrasound and histological analysis of kidneys from both Cdh16Cre::Tfebfs and Cdh16CreErt2::Tfebfs mice revealed the presence of a severe cystic disease (Figure 1E, Figure 1—figure supplement 2D and E). In Cdh16Cre::Tfebfs mice, small cysts arose mainly from the cortex and outer medulla at P12 and became significantly enlarged at P30. At P90, kidney architecture was completely disrupted by cysts (Figure 1F). Cdh16CreErt2::Tfebfs mice induced at P12 with tamoxifen and sacrificed at P90 showed a higher number of smaller cysts in both cortex and outer medulla (Figure 1F). Cysts were also observed in Cdh16CreErt2::Tfebfs induced at P14 and, to a lesser extent, at P30 (Figure 1—figure supplement 2E). Tubular epithelial cells lining the cysts showed high levels of cadherin 16, indicating the presence of Cdh16Cre-mediated Tfeb overexpression in these cells (Figure 1G). Histological analysis revealed that cysts from Cdh16Cre::Tfebfs mice were positive for AQP2 and THP and negative for megalin, indicating that they originate from collecting ducts and distal tubules and not from proximal tubules. Notably, the largest cysts were almost completely negative to all tubular markers, suggesting that they became undifferentiated. Conversely, cysts from Cdh16CreErt2::Tfebfs mice were positive to megalin and THP, indicating that they arose from proximal and distal tubules (Figure 1H, Figure 1—figure supplement 3). These differences in cyst origin have already been described in other polycystic kidney disease mouse models and have been attributed to intrinsic differences of specific renal segments at different developmental stages (Lantinga-van Leeuwen et al., 2007; Happé et al., 2009; Leonhard et al., 2016; Piontek et al., 2007).10.7554/eLife.17047.002Figure 1.Tfeb overexpressing mice display cystic kidneys.

View Article: PubMed Central - PubMed

ABSTRACT

TFE-fusion renal cell carcinomas (TFE-fusion RCCs) are caused by chromosomal translocations that lead to overexpression of the TFEB and TFE3 genes (Kauffman et al., 2014). The mechanisms leading to kidney tumor development remain uncharacterized and effective therapies are yet to be identified. Hence, the need to model these diseases in an experimental animal system (Kauffman et al., 2014). Here, we show that kidney-specific TFEB overexpression in transgenic mice, resulted in renal clear cells, multi-layered basement membranes, severe cystic pathology, and ultimately papillary carcinomas with hepatic metastases. These features closely recapitulate those observed in both TFEB- and TFE3-mediated human kidney tumors. Analysis of kidney samples revealed transcriptional induction and enhanced signaling of the WNT β-catenin pathway. WNT signaling inhibitors normalized the proliferation rate of primary kidney cells and significantly rescued the disease phenotype in vivo. These data shed new light on the mechanisms underlying TFE-fusion RCCs and suggest a possible therapeutic strategy based on the inhibition of the WNT pathway.

Doi:: http://dx.doi.org/10.7554/eLife.17047.001

No MeSH data available.


Related in: MedlinePlus