Downregulation of Dickkopf-3 disrupts prostate acinar morphogenesis through TGF-β/Smad signalling

Romero, Diana; Kawano, Yoshiaki; Bengoa, Nora; Walker, Marjorie M.; Maltry, Nicole; Niehrs, Christof; Waxman, Jonathan; Kypta, Robert M.

doi:10.1242/dev.099242

Cited by 9 publications

(21 citation statements)

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“…This study reports the unexpected cleavage of cell signaling molecules such as DKK3, which is implicated in prostate morphogenesis (26, 35) and testis development (30) probably through the regulation of TGF‐β or Wnt/β‐catenin signaling. Investigation of testis morphology in Adamts –/– male mice shows the alteration of the seminiferous tubules leading to the interesting hypothesis that processing of DKK3 by ADAMTS2 could play a role during testis development.…”

Section: Discussionmentioning

confidence: 87%

“…This analysis yielded 8, 17, and 22 potential direct substrates for ADAMTS2, ADAMTS3, and ADAMTS14 (Table 1). Besides fibrillar collagens, ADAMTS2 was found to be likely able to process the beaded filament‐forming type VI collagen, outside the triple‐helical domains; decorin, a proteoglycan interacting with the surface of collagen fibrils; and DKK3, a factor implicated in the regulation of Wnt/β‐catenin and TGF‐β signaling (25, 26). The potential direct substrates of ADAMTS3 were more numerous and included fibril‐forming collagens, other proteins of the ECM, and proteins regulating cell behavior (apoptosis and proliferation) or lipid metabolism, such as clusterin, granulin, or prosaposin (Table 1).…”

Section: Resultsmentioning

confidence: 99%

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Determination of the substrate repertoire of ADAMTS2, 3, and 14 significantly broadens their functions and identifies extracellular matrix organization and TGF‐β signaling as primary targets

et al. 2016

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A disintegrin and metalloproteinase with thrombospondin type I motif (ADAMTS)2, 3, and 14 are collectively named procollagen N-proteinases (pNPs) because of their specific ability to cleave the aminopropeptide of fibrillar procollagens. Several reports also indicate that they could be involved in other biological processes, such as blood coagulation, development, and male fertility, but the potential substrates associated with these activities remain unknown. Using the recently described N-terminal amine isotopic labeling of substrate approach, we analyzed the secretomes of human fibroblasts and identified 8, 17, and 22 candidate substrates for ADAMTS2, 3, and 14, respectively. Among these newly identified substrates, many are components of the extracellular matrix and/or proteins related to cell signaling such as latent TGF-β binding protein 1, TGF-β RIII, and dickkopf-related protein 3. Candidate substrates for the 3 ADAMTS have been biochemically validated in different contexts, and the implication of ADAMTS2 in the control of TGF-β activity has been further demonstrated in human fibroblasts. Finally, the cleavage site specificity was assessed showing a clear and unique preference for nonpolar or slightly hydrophobic amino acids. This work shows that the activities of the pNPs extend far beyond the classically reported processing of the aminopropeptide of fibrillar collagens and that they should now be considered as multilevel regulators of matrix deposition and remodeling.-Bekhouche, M., Leduc, C., Dupont, L., Janssen, L., Delolme, F., Vadon-Le Goff, S., Smargiasso, N., Baiwir, D., Mazzucchelli, G., Zanella-Cleon, I., Dubail, J., De Pauw, E., Nusgens, B., Hulmes, D. J. S., Moali, C., Colige, A. Determination of the substrate repertoire of ADAMTS2, 3, and 14 significantly broadens their functions and identifies extracellular matrix organization and TGF-β signaling as primary targets.

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Section: Discussionmentioning

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Determination of the substrate repertoire of ADAMTS2, 3, and 14 significantly broadens their functions and identifies extracellular matrix organization and TGF‐β signaling as primary targets

et al. 2016

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“…PAH-PVR is characterized by the formation of neointimal growth, the proliferation and hypertrophy of PASMCs, and deposition of ECM, including the elastin, collagen and fibronectin 10,11 . Collagen and fibronectin are the important ECM components of the vascular wall.…”

Section: ■ Discussionmentioning

confidence: 99%

Roles of the ERK1/2 and PI3K/PKB signaling pathways in regulating the expression of extracellular matrix genes in rat pulmonary artery smooth muscle cells

Jia

et al. 2017

Acta Cir. Bras.

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Roles of the ERK1/2 and PI3K/PKB signaling pathways in regulating the expression of extracellular matrix genes in rat pulmonary artery smooth muscle cells The pulmonary artery was then extracted, the fibrous tunica externa dissected and the runica intima stripped. The myogenous tissues of the vascular middle layer of the artery were then cut into 1 mm³ tissue blocks, and seeded into the culture flasks with the density as 1cm -2 . After 2 h wall-adherent cultivation at 37°C and 5% CO 2 , M199 medium containing 10% fetal bovine serum (FBS) was added. A week later, cells had grown outwards from the tissue blocks; when the growth covered 70% of the area of the base of the vessel, the culture was passaged. The 3rd-generation cells were taken for slice culturing and an immunohistochemical method was performed to detect the expression of cellular α-SMA and to confirm the purity of the PASMCs.The PASMCs were then divided into five groups and cultured in M199 medium containing different additional components, and for 48 h and 72 h. The control group was cultivated in M199 medium with 10% FBS. Indeed, in this study, the control group is negative control. The CTGF group with 10% FBS + 50 ng/ml CTGF (PeproTech, USA), the CP group with 10% FBS + 50 ng/ml CTGF + 20 μmol/L PD98059 (Sigma, USA), the CL group with 10% FBS + 50 ng/ml CTGF + 10 μmol/L LY294002 (Sigma, USA) and the CPL group with 10% FBS + 50 ng/ml CTGF + 20 μmol/L PD98059 + 10 μmol/L LY294002. RT-PCRFor the RT-PCR, the cDNA sequences of rat collagen III, fibronectin and β-actin were downloaded from NCBI gene database, then the Primer Premier 5 software was used to design appropriate PCR primers. The primer sequences were in the Table 1.

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“…The DKK3 gene product, Dickkopf-3 (Dkk-3), plays a role in prostate gland architecture [ 17 ] and displays potent tumor suppressor activity [ 18 ]. Ectopic expression of DKK3 in PCa cells leads to apoptosis [ 13 , 19 ] and inhibits proliferation, migration [ 15 , 20 ], and metastasis [ 19 ]. Moreover, injection of a DKK3-expressing adenovirus into the tumors of a patient with incurable PCa reduced metastatic tumor growth [ 21 ], and an early stage clinical trial found that intra-prostatic injection of DKK3 adenovirus significantly improved survival of patients with high-risk localized PCa [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Mediated Reactivation of DKK3 Expression Attenuates TGF-β Signaling in Prostate Cancer

Kardooni

González-Gualda

Stylianakis

et al. 2018

Cancers

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The DKK3 gene encodes a secreted protein, Dkk-3, that inhibits prostate tumor growth and metastasis. DKK3 is downregulated by promoter methylation in many types of cancer, including prostate cancer. Gene silencing studies have shown that Dkk-3 maintains normal prostate epithelial cell homeostasis by limiting TGF-β/Smad signaling. While ectopic expression of Dkk-3 leads to prostate cancer cell apoptosis, it is unclear if Dkk-3 has a physiological role in cancer cells. Here, we show that treatment of PC3 prostate cancer cells with the DNA methyltransferase (DNMT) inhibitor decitabine demethylates the DKK3 promoter, induces DKK3 expression, and inhibits TGF-β/Smad-dependent transcriptional activity. Direct induction of DKK3 expression using CRISPR-dCas9-VPR also inhibited TGF-β/Smad-dependent transcription and attenuated PC3 cell migration and proliferation. These effects were not observed in C4-2B cells, which do not respond to TGF-β. TGF-β signals can regulate gene expression directly via SMAD proteins and indirectly by increasing DNMT expression, leading to promoter methylation. Analysis of genes downregulated by promoter methylation and predicted to be regulated by TGF-β found that DKK3 induction increased expression of PTGS2, which encodes cyclooxygenase-2. Together, these observations provide support for using CRISPR-mediated induction of DKK3 as a potential therapeutic approach for prostate cancer and highlight complexities in Dkk-3 regulation of TGF-β signaling.

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Downregulation of Dickkopf-3 disrupts prostate acinar morphogenesis through TGF-β/Smad signalling

Cited by 9 publications

References 1 publication

Determination of the substrate repertoire of ADAMTS2, 3, and 14 significantly broadens their functions and identifies extracellular matrix organization and TGF‐β signaling as primary targets

Determination of the substrate repertoire of ADAMTS2, 3, and 14 significantly broadens their functions and identifies extracellular matrix organization and TGF‐β signaling as primary targets

Roles of the ERK1/2 and PI3K/PKB signaling pathways in regulating the expression of extracellular matrix genes in rat pulmonary artery smooth muscle cells

CRISPR-Mediated Reactivation of DKK3 Expression Attenuates TGF-β Signaling in Prostate Cancer

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