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Epithelial-to-mesenchymal transition induces cell cycle arrest and parenchymal damage in renal fibrosis.

Lovisa S, LeBleu VS, Tampe B, Sugimoto H, Vadnagara K, Carstens JL, Wu CC, Hagos Y, Burckhardt BC, Pentcheva-Hoang T, Nischal H, Allison JP, Zeisberg M, Kalluri R - Nat. Med. (2015)

Bottom Line: The functional consequence of the EMT program during fibrotic injury is an arrest in the G2 phase of the cell cycle and lower expression of several solute and solvent transporters in TECs.In mouse models of experimentally induced renal fibrosis, conditional deletion of Twist1 or Snai1 in proximal TECs resulted in inhibition of the EMT program and the maintenance of TEC integrity, while also restoring cell proliferation, dedifferentiation-associated repair and regeneration of the kidney parenchyma and attenuating interstitial fibrosis.Thus, inhibition of the EMT program in TECs during chronic renal injury represents a potential anti-fibrosis therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Metastasis Research Center, University of Texas M.D. Anderson Cancer Center, Houston, Texas, USA.

ABSTRACT
Kidney fibrosis is marked by an epithelial-to-mesenchymal transition (EMT) of tubular epithelial cells (TECs). Here we find that, during renal fibrosis, TECs acquire a partial EMT program during which they remain associated with their basement membrane and express markers of both epithelial and mesenchymal cells. The functional consequence of the EMT program during fibrotic injury is an arrest in the G2 phase of the cell cycle and lower expression of several solute and solvent transporters in TECs. We also found that transgenic expression of either Twist1 (encoding twist family bHLH transcription factor 1, known as Twist) or Snai1 (encoding snail family zinc finger 1, known as Snail) expression is sufficient to promote prolonged TGF-β1-induced G2 arrest of TECs, limiting the cells' potential for repair and regeneration. In mouse models of experimentally induced renal fibrosis, conditional deletion of Twist1 or Snai1 in proximal TECs resulted in inhibition of the EMT program and the maintenance of TEC integrity, while also restoring cell proliferation, dedifferentiation-associated repair and regeneration of the kidney parenchyma and attenuating interstitial fibrosis. Thus, inhibition of the EMT program in TECs during chronic renal injury represents a potential anti-fibrosis therapy.

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Inhibition of EMT reduces immune infiltration in kidney fibrosis. (a) Percentages of CD45+, CD3+, CD4+FoxP3− Teff, CD4+FoxP3+ Treg, CD8+, NK cells, CD11c+, γδ+, CD19+, CD11b+Ly6C−Ly6G−, CD11b+Ly6G+, CD11b+Ly6C+ in the indicated experimental groups. Healthy, n = 5; WT contralat., n = 11; TwistcKO contralat., n = 11; WT UUO, n = 11; TwistcKO UUO, n = 11. (b) Representative images (8 visual fields for each tissue analyzed) of immunolabelling for CD3 in the indicated experimental groups. Scale bar, 50 ~m. (c) Quantification of the number of CD3+ cells per visual field in the indicated experimental groups. WT contralat., n = 4 ; TwistcKO contralat., n = 4; WT UUO, n = 4; TwistcKO UUO, n = 4. For panel a data is presented as mean ± SD; for c data is presented as mean ± SEM. One–way ANOVA with Tukey post–hoc analysis was used. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. contralat.: contralateral kidney.
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Figure 4: Inhibition of EMT reduces immune infiltration in kidney fibrosis. (a) Percentages of CD45+, CD3+, CD4+FoxP3− Teff, CD4+FoxP3+ Treg, CD8+, NK cells, CD11c+, γδ+, CD19+, CD11b+Ly6C−Ly6G−, CD11b+Ly6G+, CD11b+Ly6C+ in the indicated experimental groups. Healthy, n = 5; WT contralat., n = 11; TwistcKO contralat., n = 11; WT UUO, n = 11; TwistcKO UUO, n = 11. (b) Representative images (8 visual fields for each tissue analyzed) of immunolabelling for CD3 in the indicated experimental groups. Scale bar, 50 ~m. (c) Quantification of the number of CD3+ cells per visual field in the indicated experimental groups. WT contralat., n = 4 ; TwistcKO contralat., n = 4; WT UUO, n = 4; TwistcKO UUO, n = 4. For panel a data is presented as mean ± SD; for c data is presented as mean ± SEM. One–way ANOVA with Tukey post–hoc analysis was used. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. contralat.: contralateral kidney.

Mentions: Inflammation has been widely described to be a key driver of kidney fibrosis42. Initiated as a protective response to injury, the immune infiltrate evolves and may contribute to fibrotic scarring, leading to the loss of functional parenchyma. We assayed by flow cytometry and immunostaining the differential composition of the immune infiltrate in WT and TwistcKO UUO kidneys compared to contralateral and to kidneys from healthy mice. While contralateral and healthy kidneys showed similar immune infiltrate (Fig. 4a–c), UUO kidneys showed a significant increase in CD45+ leukocyte infiltration (Fig. 4a). The percent CD3+ T cells, both effector (CD4+Foxp3−) and regulator (CD4+Foxp3+) T cells, cytotoxic CD8+ T cells, natural killer cells (NK1.1+), gamma delta (γδ+) T cells, CD11c+ dentritic cells, and CD19+ B cells were significantly increased in fibrotic kidneys compared to contralateral kidneys in WT mice (Fig. 4a).


Epithelial-to-mesenchymal transition induces cell cycle arrest and parenchymal damage in renal fibrosis.

Lovisa S, LeBleu VS, Tampe B, Sugimoto H, Vadnagara K, Carstens JL, Wu CC, Hagos Y, Burckhardt BC, Pentcheva-Hoang T, Nischal H, Allison JP, Zeisberg M, Kalluri R - Nat. Med. (2015)

Inhibition of EMT reduces immune infiltration in kidney fibrosis. (a) Percentages of CD45+, CD3+, CD4+FoxP3− Teff, CD4+FoxP3+ Treg, CD8+, NK cells, CD11c+, γδ+, CD19+, CD11b+Ly6C−Ly6G−, CD11b+Ly6G+, CD11b+Ly6C+ in the indicated experimental groups. Healthy, n = 5; WT contralat., n = 11; TwistcKO contralat., n = 11; WT UUO, n = 11; TwistcKO UUO, n = 11. (b) Representative images (8 visual fields for each tissue analyzed) of immunolabelling for CD3 in the indicated experimental groups. Scale bar, 50 ~m. (c) Quantification of the number of CD3+ cells per visual field in the indicated experimental groups. WT contralat., n = 4 ; TwistcKO contralat., n = 4; WT UUO, n = 4; TwistcKO UUO, n = 4. For panel a data is presented as mean ± SD; for c data is presented as mean ± SEM. One–way ANOVA with Tukey post–hoc analysis was used. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. contralat.: contralateral kidney.
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Figure 4: Inhibition of EMT reduces immune infiltration in kidney fibrosis. (a) Percentages of CD45+, CD3+, CD4+FoxP3− Teff, CD4+FoxP3+ Treg, CD8+, NK cells, CD11c+, γδ+, CD19+, CD11b+Ly6C−Ly6G−, CD11b+Ly6G+, CD11b+Ly6C+ in the indicated experimental groups. Healthy, n = 5; WT contralat., n = 11; TwistcKO contralat., n = 11; WT UUO, n = 11; TwistcKO UUO, n = 11. (b) Representative images (8 visual fields for each tissue analyzed) of immunolabelling for CD3 in the indicated experimental groups. Scale bar, 50 ~m. (c) Quantification of the number of CD3+ cells per visual field in the indicated experimental groups. WT contralat., n = 4 ; TwistcKO contralat., n = 4; WT UUO, n = 4; TwistcKO UUO, n = 4. For panel a data is presented as mean ± SD; for c data is presented as mean ± SEM. One–way ANOVA with Tukey post–hoc analysis was used. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. contralat.: contralateral kidney.
Mentions: Inflammation has been widely described to be a key driver of kidney fibrosis42. Initiated as a protective response to injury, the immune infiltrate evolves and may contribute to fibrotic scarring, leading to the loss of functional parenchyma. We assayed by flow cytometry and immunostaining the differential composition of the immune infiltrate in WT and TwistcKO UUO kidneys compared to contralateral and to kidneys from healthy mice. While contralateral and healthy kidneys showed similar immune infiltrate (Fig. 4a–c), UUO kidneys showed a significant increase in CD45+ leukocyte infiltration (Fig. 4a). The percent CD3+ T cells, both effector (CD4+Foxp3−) and regulator (CD4+Foxp3+) T cells, cytotoxic CD8+ T cells, natural killer cells (NK1.1+), gamma delta (γδ+) T cells, CD11c+ dentritic cells, and CD19+ B cells were significantly increased in fibrotic kidneys compared to contralateral kidneys in WT mice (Fig. 4a).

Bottom Line: The functional consequence of the EMT program during fibrotic injury is an arrest in the G2 phase of the cell cycle and lower expression of several solute and solvent transporters in TECs.In mouse models of experimentally induced renal fibrosis, conditional deletion of Twist1 or Snai1 in proximal TECs resulted in inhibition of the EMT program and the maintenance of TEC integrity, while also restoring cell proliferation, dedifferentiation-associated repair and regeneration of the kidney parenchyma and attenuating interstitial fibrosis.Thus, inhibition of the EMT program in TECs during chronic renal injury represents a potential anti-fibrosis therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Metastasis Research Center, University of Texas M.D. Anderson Cancer Center, Houston, Texas, USA.

ABSTRACT
Kidney fibrosis is marked by an epithelial-to-mesenchymal transition (EMT) of tubular epithelial cells (TECs). Here we find that, during renal fibrosis, TECs acquire a partial EMT program during which they remain associated with their basement membrane and express markers of both epithelial and mesenchymal cells. The functional consequence of the EMT program during fibrotic injury is an arrest in the G2 phase of the cell cycle and lower expression of several solute and solvent transporters in TECs. We also found that transgenic expression of either Twist1 (encoding twist family bHLH transcription factor 1, known as Twist) or Snai1 (encoding snail family zinc finger 1, known as Snail) expression is sufficient to promote prolonged TGF-β1-induced G2 arrest of TECs, limiting the cells' potential for repair and regeneration. In mouse models of experimentally induced renal fibrosis, conditional deletion of Twist1 or Snai1 in proximal TECs resulted in inhibition of the EMT program and the maintenance of TEC integrity, while also restoring cell proliferation, dedifferentiation-associated repair and regeneration of the kidney parenchyma and attenuating interstitial fibrosis. Thus, inhibition of the EMT program in TECs during chronic renal injury represents a potential anti-fibrosis therapy.

Show MeSH
Related in: MedlinePlus