Metadata supporting data files of the related article: Wnt5a Induces ROR1 To Recruit Cortactin To Promote Breast Cancer Migration And Metastasis
Creators: name: Md Kamrul Hasan
name: Widhopf, George F.
name: Suping Zhang
name: Lam, Sharon M
name: Zhouxin Shen
name: Briggs, Steven P.
name: Parker, Barbara A.
name: Kipps, Thomas J.
Publisher: name: figshare
Year: 2019
Subjects: subject: Cancer
subject: Cell Biology
subject: Medicine
subject: 60109 Proteomics and Intermolecular Interactions (excl. Medical Proteomics)
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: 111201 Cancer Cell Biology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: 111299 Oncology and Carcinogenesis not elsewhere classified

description: The study examined whether the Wnt5a protein can stimulate the tyrosine-protein kinase transmembrane receptor ROR1 to complex with cortactin and thereby promote breast cancer-cell migration/metastasis.<br><b>Data access:</b> Datasets Association figure1.xlsx, Activation, migration figure2.xlsx, Migration figure3.xlsx, Metastasis figure4.xlsx and Characteristics table1.xlsx are not publicly available to protect patient privacy, but can be accessed from the corresponding author on request, upon the completion of a Data Usage Agreement. Corresponding author details: <b>Dr. Thomas Kipps</b>, Moores Cancer Center, University of California, San Diego, 3855 Health Sciences Drive, Rm 4307, La Jolla, California 92093-0820, USA.; E-mail: <b>tkipps@ucsd.edu</b>. Raw breast cancer PDX-derived mass spectrometry data (supporting figure 1, and identifying cortactin as the protein that associates with ROR1), are publicly available in the Japan ProteOme STandard Repository (<b>jPOST repository</b>) under the accession ID: <b>JPST000678</b>. All the uncropped Western blots generated during this study are available in Supplementary Figure S11.<br><b>Study approval: </b>Primary breast-tumor specimens were collected from patients, who provided written informed consent on a protocol approved by the Institutional Review Board of the University of California, San Diego (approval number 090401), in accordance with the Declaration of Helsinki. The animal study protocol was approved by the University of California San Diego Institutional Animal Care and Use Committee (approval number S03037).<br><b>Study aims and methodology:</b><b><br></b>The present study aimed to examine a previously unrecognized ROR1/cortactin/ARHGEF1-dependent pathway leading to activation of RhoA in response to Wnt5a in breast cancer cells, that promotes breast cancer-cell migration/metastasis.<br>The following methods are described in detail in the published article: immunoblot analysis, metastasis assay and statistical analysis.The following methods are described in detail in the “supplementary materials and methods” of the related article: mass spectrometry analysis, cell culture of MCF7 cells, MCF7 transfectants and breast cancer patient-derived xenograft (PDX) cells, immunoprecipitation analysis, nucleofection of plasmids and siRNA, RhoGEF nucleotide exchange activity assay, RhoA activation assay, flow cytometry analysis, site-directed mutagenesis, cell migration assay, animal experiments (6-week-old female Rag2-/-γc-/- mice were used in this study) and PDX models. <br><b>Dataset description:</b><b><br></b><b>Data supporting figure 1: </b>Dataset <b>Association figure1.xlsx </b>is in <b>.xlsx</b> file format and includes immunoblot analyses of anti-ROR1 or anti-cortactin immune precipitates in breast-cancer PDX cells, and immunoblot analyses of lysates prepared from serum-starved breast cancer PDX cells that subsequently were treated with Ctrl-IgG or cirmtuzumab, without or with Wnt5a.<br><b>Data supporting figure 2: </b>Dataset <b>Activationmigration figure2.xlsx</b> is in <b>.xlsx</b> file format and includes immunoblot analyses data on the ROR1-dependent phosphorylation of cortactin in breast cancer PDX cells.<br><b>Data supporting figure 3: </b>Dataset <b>Migration figure3.xlsx</b> is in .<b>xlsx</b> file format and includes data on the interaction of ROR1 with cortactin or ARHGEF1 in the presence of Wnt5a by immunoblot analysis of anti-ROR1 immune precipitates from lysates of MCF7-control (Ctrl), MCF7-ΔPRD (ROR1 without its proline rich region), MCF7-wild type (WT) ROR1 or MCF7 cells transfected with each of the various mutated forms of ROR1.<br><b>Data supporting figure 4:</b> Dataset <b>Metastasis figure4.xlsx </b>is in <b>.xlsx</b> file format and includes data on the metastatic capacity of MCF7 cells transfected with different ROR1 constructs (various mutated forms of ROR1).<br><b>Data supporting table 1: </b>Dataset <b>Characteristics table1.xlsx</b> is in <b>.xlsx</b> file format and includes characteristics of the breast cancer patient-derived xenograft (PDX) samples<br>