Volume 15, Issue 11, Pages (June 2016)

Slides:



Advertisements
Similar presentations
Volume 14, Issue 10, Pages (March 2016)
Advertisements

Volume 17, Issue 5, Pages (May 2015)
Volume 19, Issue 5, Pages (May 2017)
Knockdown of Bone Morphogenetic Proteins Type 1a Receptor (BMPR1a) in Breast Cancer Cells Protects Bone from Breast Cancer-Induced Osteolysis by Suppressing.
Volume 24, Issue 5, Pages (November 2013)
Volume 21, Issue 13, Pages (December 2017)
Volume 23, Issue 4, Pages (April 2018)
Foxf2 in Intestinal Fibroblasts Reduces Numbers of Lgr5+ Stem Cells and Adenoma Formation by Inhibiting Wnt Signaling   Ali Moussavi Nik, Azadeh Reyahi,
Up-Regulation of RFC3 Promotes Triple Negative Breast Cancer Metastasis and is Associated With Poor Prognosis Via EMT  Zhen-Yu He, San-Gang Wu, Fang Peng,
Modulation of K-Ras-Dependent Lung Tumorigenesis by MicroRNA-21
Silencing of Discoidin Domain Receptor-1 (DDR1) Concurrently Inhibits Multiple Steps of Metastasis Cascade in Gastric Cancer  Ryo Yuge, Yasuhiko Kitadai,
Stromal Fibroblast–Specific Expression of ADAM-9 Modulates Proliferation and Apoptosis in Melanoma Cells In Vitro and In Vivo  Anna N. Abety, Jay W. Fox,
CCN2 Expression by Tumor Stroma Is Required for Melanoma Metastasis
Combined PLX3397 and PTX treatment inhibits metastasis in a CD8-dependent manner. Combined PLX3397 and PTX treatment inhibits metastasis in a CD8-dependent.
Pulmonary Fibroblasts Induce Epithelial Mesenchymal Transition and Some Characteristics of Stem Cells in Non-Small Cell Lung Cancer  Yasushi Shintani,
Volume 18, Issue 14, Pages (July 2008)
Volume 6, Issue 5, Pages (March 2014)
Volume 12, Issue 2, Pages (August 2007)
Volume 21, Issue 1, Pages (January 2012)
Regulation of Mammary Luminal Cell Fate and Tumorigenesis by p38α
Fibroblast-Derived MMP-14 Regulates Collagen Homeostasis in Adult Skin
Volume 20, Issue 6, Pages (December 2011)
Manuela Schmidt, Danny Gutknecht, Jan C
Volume 24, Issue 13, Pages e5 (September 2018)
Volume 25, Issue 6, Pages (June 2017)
Volume 29, Issue 4, Pages (April 2016)
LATS2 is a tumor suppressor in a mouse lumB breast cancer model.
TGF-β-Induced (TGFBI) Protein in Melanoma: A Signature of High Metastatic Potential  Laura Lauden, Johan Siewiera, Wahid Boukouaci, Kiran Ramgolam, Samia.
Volume 14, Issue 10, Pages (March 2016)
Volume 133, Issue 4, Pages (May 2008)
Cancer-Associated Fibroblasts Are Activated in Incipient Neoplasia to Orchestrate Tumor-Promoting Inflammation in an NF-κB-Dependent Manner  Neta Erez,
Hosein Kouros-Mehr, Euan M. Slorach, Mark D. Sternlicht, Zena Werb 
Melanoma Suppressor Functions of the Carcinoma Oncogene FOXQ1
Volume 5, Issue 5, Pages (December 2013)
Tumor Self-Seeding by Circulating Cancer Cells
Volume 117, Issue 7, Pages (June 2004)
Volume 16, Issue 2, Pages (July 2016)
Kimberly M. McDermott, Bob Y. Liu, Thea D. Tlsty, Gregory J. Pazour 
Paola Zigrino, Isolde Kuhn, Tobias Bäuerle, Jan Zamek, Jay W
Overexpression of CD109 in the Epidermis Differentially Regulates ALK1 Versus ALK5 Signaling and Modulates Extracellular Matrix Synthesis in the Skin 
IL-22 Promotes Fibroblast-Mediated Wound Repair in the Skin
Volume 24, Issue 4, Pages (July 2018)
Collagen VII Half-Life at the Dermal-Epidermal Junction Zone: Implications for Mechanisms and Therapy of Genodermatoses  Tobias Kühl, Markus Mezger, Ingrid.
14-3-3σ Regulates Keratinocyte Proliferation and Differentiation by Modulating Yap1 Cellular Localization  Sumitha A.T. Sambandam, Ramesh B. Kasetti,
Volume 6, Issue 5, Pages (May 2016)
Rita U. Lukacs, Sanaz Memarzadeh, Hong Wu, Owen N. Witte 
Volume 14, Issue 10, Pages (March 2016)
Volume 8, Issue 6, Pages (December 2010)
Volume 16, Issue 5, Pages (August 2016)
Volume 13, Issue 11, Pages (December 2015)
Volume 9, Issue 4, Pages (October 2017)
Hepsin promotes prostate cancer progression and metastasis
Increased Expression of Wnt2 and SFRP4 in Tsk Mouse Skin: Role of Wnt Signaling in Altered Dermal Fibrillin Deposition and Systemic Sclerosis  Julie Bayle,
Volume 7, Issue 1, Pages (January 2008)
Volume 8, Issue 3, Pages (August 2014)
TAK1 Is Required for Dermal Wound Healing and Homeostasis
Volume 21, Issue 1, Pages (January 2013)
(A) Schematic representation of the conditional Lats2 locus.
Volume 24, Issue 12, Pages (September 2018)
Volume 26, Issue 11, Pages e5 (March 2019)
Volume 11, Issue 3, Pages (April 2015)
Volume 25, Issue 11, Pages e6 (December 2018)
Cellular Heterogeneity in the Mouse Esophagus Implicates the Presence of a Nonquiescent Epithelial Stem Cell Population  Aaron D. DeWard, Julie Cramer,
Volume 8, Issue 4, Pages (April 2017)
Kristiina Airola, Norbert E. Fusenig 
Effective Therapy Using a Liposomal siRNA that Targets the Tumor Vasculature in a Model Murine Breast Cancer with Lung Metastasis  Yu Sakurai, Tomoya.
Volume 15, Issue 4, Pages (April 2016)
Volume 12, Issue 2, Pages (August 2007)
Foxf2 in Intestinal Fibroblasts Reduces Numbers of Lgr5+ Stem Cells and Adenoma Formation by Inhibiting Wnt Signaling   Ali Moussavi Nik, Azadeh Reyahi,
Volume 2, Issue 6, Pages (December 2012)
Presentation transcript:

Volume 15, Issue 11, Pages 2510-2523 (June 2016) The Action of Discoidin Domain Receptor 2 in Basal Tumor Cells and Stromal Cancer- Associated Fibroblasts Is Critical for Breast Cancer Metastasis  Callie A.S. Corsa, Audrey Brenot, Whitney R. Grither, Samantha Van Hove, Andrew J. Loza, Kun Zhang, Suzanne M. Ponik, Yuming Liu, David G. DeNardo, Kevin W. Eliceiri, Patricia J. Keely, Gregory D. Longmore  Cell Reports  Volume 15, Issue 11, Pages 2510-2523 (June 2016) DOI: 10.1016/j.celrep.2016.05.033 Copyright © 2016 The Author(s) Terms and Conditions

Cell Reports 2016 15, 2510-2523DOI: (10.1016/j.celrep.2016.05.033) Copyright © 2016 The Author(s) Terms and Conditions

Figure 1 Generation and Characterization of Modified Mouse DDR2 Alleles (A) Schematic of Ddr2 alleles. Ddr2null mice were crossed with β-actin-Cre mice to generate the Ddr2lacz allele. Ddr2null mice were crossed with FLPO mice to generate the conditional floxed allele (Ddr2fl). (B) Body weights of various mice, calculated as percentage of WT littermate. ∗p < 0.001, ten mice per group. Data are presented as mean ± SD. (C) Representative images confirming the dwarfism phenotype in Ddr2−/− mice. (D) Western blot of extracts from dermal fibroblasts isolated from Ddr2+/+ and Ddr2null/null mice with indicated antibodies. (E) PCR detection of floxed and recombined DDR2 alleles in dermal fibroblasts isolated from indicated mice. (F) Western blot of extracts from dermal fibroblasts isolated from indicated mice, with the indicated antibodies. Cell Reports 2016 15, 2510-2523DOI: (10.1016/j.celrep.2016.05.033) Copyright © 2016 The Author(s) Terms and Conditions

Figure 2 Ubiquitous Deletion of DDR2 Reduces Breast Cancer Lung Metastasis without Affecting Primary Tumor Growth (A) Primary MMTV-PyMT tumor growth rates in indicated mice, determined by the age at which the largest tumor reached 2 cm in diameter. 10–40 mice per group. Data are presented as mean ± SD. (B) Total tumor burden of indicated mice, determined by the sum of the volume of all tumors for each mouse when the largest tumor reached 2 cm in diameter. Tumor volumes were calculated by V = 0.52 × (W2) × L. 10–40 mice per group. Data are presented as mean ± SD. (C) Quantification of number of lung metastases in indicated mice calculated by the average number of microscopically visible metastases counted from three H&E-stained sections of lung from each mouse taken 200 μm apart and normalized to the total lung area. ∗p < 0.01; N.S., not significant. 10–40 mice per group. Data are presented as mean ± SD. Cell Reports 2016 15, 2510-2523DOI: (10.1016/j.celrep.2016.05.033) Copyright © 2016 The Author(s) Terms and Conditions

Figure 3 DDR2 Expression in Mouse MMTV-PyMT Breast Tumor Development and Progression (A) X-gal staining of mammary gland/tumor sections from Ddr2lacZ/+; MMTV-PyMT mice at multiple times during tumor development. Scale bars, 50 μm. (B) X-gal staining on hyperplastic mammary glands from 5-week-old Ddr2lacZ/+; MMTV-PyMT mice. Scale bars, 50 μm. Arrowhead indicates stromal cell, black arrow indicates luminal epithelial cell, and red arrow indicates basal myoepithelial cell. (C) X-gal staining and immunofluorescence for K14 (yellow), K8 (green), and α-SMA (red) on sequential tissue slices from hyperplastic mammary glands: 5-week-old Ddr2lacZ/+; MMTV-PyMT mice. Scale bars, 50 μm. (D) Dual in situ hybridization for mRNA (left) of 13-week primary breast tumor slices from Ddr2+/+; MMTV-PyMT mice. K14 (red) and DDR2 (green) stain in the top panels and K8 (red) and DDR2 (green) stain in the bottom panels. White arrows on H&E-stained images (right) identify cells expressing both transcripts. Scale bars, 50 μm. Cell Reports 2016 15, 2510-2523DOI: (10.1016/j.celrep.2016.05.033) Copyright © 2016 The Author(s) Terms and Conditions

Figure 4 DDR2 Deletion in Basal, but Not Luminal, Breast Epithelial Cells Inhibit Lung Metastasis (A) Primary tumor growth rates in indicated mice, determined by the age at which the largest tumor reached 2 cm in diameter. 15–35 mice per group. Data are presented as mean ± SD. (B) Total tumor burden of indicated mice, determined by the sum of the volume of all tumors for each mouse when the largest tumor reached 2 cm in diameter. Tumor volumes were calculated by V = 0.52 ⋅ (Wˆ2) ⋅ L. 15–35 mice per group. Data are presented as mean ± SD. (C) Quantification of lung metastases in indicated mice presented as the average number of microscopically visible metastases counted from three H&E-stained sections of lung from each mouse taken 200 μm apart, normalized to the total lung area. ∗p < 0.01; N.S., not significant. 15–35 mice per group. Data are presented as mean ± SD. Cell Reports 2016 15, 2510-2523DOI: (10.1016/j.celrep.2016.05.033) Copyright © 2016 The Author(s) Terms and Conditions

Figure 5 The Action of DDR2 in the Tumor Stroma Is Critical for Metastasis (A and B) Reciprocal transplant of MMTV-PyMT primary tumors cells from the indicated genotypes were transplanted into the breasts of syngeneic (FVBn) hosts of the indicated genotype. Time to the development of a 2-cm breast tumor was scored (A). 8–11 mice per group. Quantification of lung metastases as the average number of microscopically visible metastases counted from three H&E-stained sections of lung from each mouse taken 200 μm apart normalized to the total lung area (B). ∗p < 0.01; N.S., not significant. 8–11 mice per group. Data are presented as mean ± SD. (C) Picrosirius red staining on tumor sections from Ddr2+/+; MMTV-PyMT and ubiquitous Ddr2−/−; MMTV-PyMT mice. Scale bars, 100 μm. (D) Quantification of Picrosirius red staining from (C). Positive staining was calculated as the percentage of red pixels over the total number of pixels in the image. ∗p < 0.001. Data are presented as mean ± SD. (E) Representative second-harmonic-generation images of collagen organization in 10- to 13-week tumors from Ddr2+/+ and ubiquitous Ddr2−/− mice. Curly collagen fibers (left image) were scored as TACS-1, and straight collagen fibers (right image) were scored as TACS-2/3. (F) Percentage of images scored as TACS-1 or TACS-2/3 from Ddr2+/+ PyMT tumors and Ddr2−/− PyMT tumors. Three 13-week and four 10-week mice of each genotype were analyzed. Three or four tumors from each mouse were isolated, and three to four images from each tumor were acquired. Total of 77 images scored. Student’s t test, ∗p < 0.05. Data are presented as mean ± SD. Cell Reports 2016 15, 2510-2523DOI: (10.1016/j.celrep.2016.05.033) Copyright © 2016 The Author(s) Terms and Conditions

Figure 6 The Action of DDR2 in CAFs Regulates Production of Tumor-Associated ECM (A) Western blot for DDR2, SNAIL1, α-SMA, and FSP1 proteins in normal human breast fibroblasts or human breast CAFs. (B) Dual in situ hybridization for mRNA (left) of 13-week primary breast tumor slices from Ddr2+/+; MMTV-PyMT mice. FAP (red) and DDR2 (green). White arrows on H&E-stained images (right) identify cells within the tumor stroma that express both transcripts. White arrowheads identify tumor cells expressing DDR2 mRNA. Scale bars, 50 μm. (C) Immunofluorescence staining for collagen 1α1 matrix produced by mouse breast tumor CAFs. Scale bars, 50 μm. (D) Quantification of global fiber alignment in (C). Data are presented as mean ± SD. (E) Immunofluorescence staining for fibronectin matrix produced by normal human mammary fibroblasts (left), human breast tumor CAFs (control SCR shRNA; middle), or hCAFs depleted of DDR2 by shRNA (right). Scale bars, 50 μm. (F) Quantification of global fiber alignment in (E). Data are presented as mean ± SD. (G) qPCR analysis of mRNA isolated from human CAFs, short hairpin (sh) RNA (shRNA) depleted of DDR2, or transduced with scrambled control (SCR). Multiple replicates of each gene were performed for each experiment. Data are representative of two independent experiments. Data are presented as mean ± SD. ∗p < 0.01. Cell Reports 2016 15, 2510-2523DOI: (10.1016/j.celrep.2016.05.033) Copyright © 2016 The Author(s) Terms and Conditions

Figure 7 Action of DDR2 in K14+ Tumor Epithelial Cells and CAFS Regulate Collective Invasion by Primary Tumor Organoids (A-D) Representative images of tumor organoids scored. (A) Non-invasive tumor organoid, no added CAFs. (B) Invasive WT tumor organoid, no added CAFs. (C) Invasive WT tumor organoid cultured in the presence of WT breast CAFs. (D) Invasive WT tumor organoid cultured in the presence of Ddr2−/− breast CAFs. Scale bars, 100 μm. (E) Quantification of number of invasive tumor organoids as a percentage of total organoids scored. In each well, 30 organoids were scored. All experimental conditions were performed in triplicate. The experiment was performed three separate times. ∗∗p < 0.001; NS, no significant difference. Data are presented as mean ± SD. Cell Reports 2016 15, 2510-2523DOI: (10.1016/j.celrep.2016.05.033) Copyright © 2016 The Author(s) Terms and Conditions