WNT4 Regulates Cellular Metabolism via Intracellular Activity at the Mitochondria in Breast and Gynecologic Cancers

Wnt ligand WNT4 is critical in female reproductive tissue development, with WNT4 dysregulation linked to related pathologies including breast cancer (invasive lobular carcinoma, ILC) and gynecologic cancers. WNT4 signaling in these contexts is distinct from canonical Wnt signaling yet inadequately understood. We previously identified atypical intracellular activity of WNT4 (independent of Wnt secretion) regulating mitochondrial function, and herein examine intracellular functions of WNT4. We further examine how convergent mechanisms of WNT4 dysregulation impact cancer metabolism. In ILC, WNT4 is co-opted by estrogen receptor α (ER) via genomic binding in WNT4 intron 1, while in gynecologic cancers, a common genetic polymorphism (rs3820282) at this ER binding site alters WNT4 regulation. Using proximity biotinylation (BioID), we show canonical Wnt ligand WNT3A is trafficked for secretion, but WNT4 is localized to the cytosol and mitochondria. We identified DHRS2, mTOR, and STAT1 as putative WNT4 cytosolic/mitochondrial signaling partners. Whole metabolite profiling, and integrated transcriptomic data, support that WNT4 mediates metabolic reprogramming via fatty acid and amino acid metabolism. Further, ovarian cancer cell lines with rs3820282 variant genotype are WNT4-dependent and have active WNT4 metabolic signaling. In protein array analyses of a cohort of 103 human gynecologic tumors enriched for patient diversity, germline rs3820282 genotype is associated with metabolic remodeling. Variant genotype tumors show increased AMPK activation and downstream signaling, with the highest AMPK signaling activity in variant genotype tumors from non-White patients. Taken together, atypical intracellular WNT4 signaling, in part via genetic dysregulation, regulate the distinct metabolic phenotypes of ILC and gynecologic cancers. Significance WNT4 regulates breast and gynecologic cancer metabolism via a previously unappreciated intracellular signaling mechanism at the mitochondria, with WNT4 mediating metabolic remodeling. Understanding WNT4 dysregulation by estrogen and genetic polymorphism offers new opportunities for defining tumor biology, precision therapeutics, and personalized cancer risk assessment.

[1]  Günter P. Wagner,et al.  A SNP affects Wnt4 expression in endometrial stroma, with antagonistic implications for pregnancy, endometriosis and reproductive cancers , 2022, bioRxiv.

[2]  J. Xia,et al.  Using MetaboAnalyst 5.0 for LC–HRMS spectra processing, multi-omics integration and covariate adjustment of global metabolomics data , 2022, Nature Protocols.

[3]  G. Hortobagyi,et al.  Invasive lobular carcinoma: an understudied emergent subtype of breast cancer , 2022, Breast Cancer Research and Treatment.

[4]  Brad T. Sherman,et al.  DAVID: a web server for functional enrichment analysis and functional annotation of gene lists (2021 update) , 2022, Nucleic Acids Res..

[5]  B. Katzenellenbogen,et al.  Targeting metabolic adaptations in the breast cancer–liver metastatic niche using dietary approaches to improve endocrine therapy efficacy , 2021, bioRxiv.

[6]  Kathleen M. Jagodnik,et al.  Gene Set Knowledge Discovery with Enrichr , 2021, Current protocols.

[7]  J. Costello,et al.  Mediator of DNA damage checkpoint 1 (MDC1) is a novel estrogen receptor co-regulator in invasive lobular carcinoma of the breast , 2020, bioRxiv.

[8]  S. Oesterreich,et al.  Estrogen Regulation of mTOR Signaling and Mitochondrial Function in Invasive Lobular Carcinoma Cell Lines Requires WNT4 , 2020, Cancers.

[9]  G. Raj,et al.  Riluzole Suppresses Growth and Enhances Response to Endocrine Therapy in ER+ Breast Cancer , 2020, bioRxiv.

[10]  Sarat Chandarlapaty,et al.  Head-to-Head Evaluation of 18F-FES and 18F-FDG PET/CT in Metastatic Invasive Lobular Breast Cancer , 2020, The Journal of Nuclear Medicine.

[11]  Anup D Shah,et al.  LFQ-Analyst: An easy-to-use interactive web-platform to analyze and visualize label-free proteomics data preprocessed with MaxQuant. , 2020, Journal of proteome research.

[12]  Benjamin G. Bitler,et al.  Wnt family member 4 (WNT4) and WNT3A activate cell-autonomous Wnt signaling independent of porcupine O-acyltransferase or Wnt secretion , 2019, The Journal of Biological Chemistry.

[13]  Paul Flicek,et al.  The International Genome Sample Resource (IGSR) collection of open human genomic variation resources , 2019, Nucleic Acids Res..

[14]  Benjamin G. Bitler,et al.  Activation of Wnt signaling promotes olaparib resistant ovarian cancer , 2019, Molecular carcinogenesis.

[15]  Sky W. Brubaker,et al.  Multi-Omic Approaches Identify Metabolic and Autophagy Regulators Important in Ovarian Cancer Dissemination , 2019, iScience.

[16]  Xiangjian Luo,et al.  DHRS2 mediates cell growth inhibition induced by Trichothecin in nasopharyngeal carcinoma , 2019, Journal of experimental & clinical cancer research : CR.

[17]  O. Frings,et al.  SubCellBarCode: Proteome-wide Mapping of Protein Localization and Relocalization. , 2019, Molecular cell.

[18]  Damian Szklarczyk,et al.  STRING v11: protein–protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets , 2018, Nucleic Acids Res..

[19]  A. D’Alessandro,et al.  Inhibition of Amino Acid Metabolism Selectively Targets Human Leukemia Stem Cells. , 2018, Cancer cell.

[20]  B. Van Houten,et al.  Key regulators of lipid metabolism drive endocrine resistance in invasive lobular breast cancer , 2018, Breast cancer research : BCR.

[21]  B. Győrffy,et al.  Integrated molecular analysis of Tamoxifen-resistant invasive lobular breast cancer cells identifies MAPK and GRM/mGluR signaling as therapeutic vulnerabilities , 2018, Molecular and Cellular Endocrinology.

[22]  Adrian V. Lee,et al.  Invasive lobular and ductal breast carcinoma differ in immune response, protein translation efficiency and metabolism , 2018, Scientific Reports.

[23]  Xiaonan Liu,et al.  An AP-MS- and BioID-compatible MAC-tag enables comprehensive mapping of protein interactions and subcellular localizations , 2018, Nature Communications.

[24]  Daniel E. Miller,et al.  Genetic Associations With Gestational Duration and Spontaneous Preterm Birth , 2018, Obstetric Anesthesia Digest.

[25]  Phillip G. Montgomery,et al.  Defining a Cancer Dependency Map , 2017, Cell.

[26]  G. Hospers,et al.  18F-FES PET Has Added Value in Staging and Therapy Decision Making in Patients With Disseminated Lobular Breast Cancer. , 2017, Clinical nuclear medicine.

[27]  S. Oesterreich,et al.  Distinct Pattern of Metastases in Patients with Invasive Lobular Carcinoma of the Breast , 2017, Geburtshilfe und Frauenheilkunde.

[28]  J. Koo,et al.  Expression of Lipid Metabolism-Related Proteins Differs between Invasive Lobular Carcinoma and Invasive Ductal Carcinoma , 2017, International journal of molecular sciences.

[29]  Adrian V. Lee,et al.  WNT4 mediates estrogen receptor signaling and endocrine resistance in invasive lobular carcinoma cell lines , 2016, Breast Cancer Research.

[30]  M. Gönen,et al.  Initial Results of a Prospective Clinical Trial of 18F-Fluciclovine PET/CT in Newly Diagnosed Invasive Ductal and Invasive Lobular Breast Cancers , 2016, The Journal of Nuclear Medicine.

[31]  R. Bernards,et al.  Integration of genomic, transcriptomic and proteomic data identifies two biologically distinct subtypes of invasive lobular breast cancer , 2016, Scientific Reports.

[32]  T. Harris,et al.  The Signal Transducer and Activator of Transcription 1 (STAT1) Inhibits Mitochondrial Biogenesis in Liver and Fatty Acid Oxidation in Adipocytes , 2015, PloS one.

[33]  Steven J. M. Jones,et al.  Comprehensive Molecular Portraits of Invasive Lobular Breast Cancer , 2015, Cell.

[34]  K. Hess,et al.  Progesterone and Overlooked Endocrine Pathways in Breast Cancer Pathogenesis. , 2015, Endocrinology.

[35]  Yurii B. Shvetsov,et al.  Identification of six new susceptibility loci for invasive epithelial ovarian cancer , 2015, Nature Genetics.

[36]  W. Jung,et al.  Expression of metabolism-related proteins in invasive lobular carcinoma: comparison to invasive ductal carcinoma , 2014, Tumor Biology.

[37]  D. Dabbs,et al.  Invasive lobular carcinoma cell lines are characterized by unique estrogen-mediated gene expression patterns and altered tamoxifen response. , 2014, Cancer research.

[38]  A. Larner,et al.  Toward a new STATe: the role of STATs in mitochondrial function. , 2014, Seminars in immunology.

[39]  R. Knight,et al.  Signal transducer and activator of transcription-1 localizes to the mitochondria and modulates mitophagy , 2013, JAK-STAT.

[40]  Amber L. Couzens,et al.  The CRAPome: a Contaminant Repository for Affinity Purification Mass Spectrometry Data , 2013, Nature Methods.

[41]  C. Alexander,et al.  Wnt signaling in mammary glands: plastic cell fates and combinatorial signaling. , 2012, Cold Spring Harbor perspectives in biology.

[42]  R. López-López,et al.  Proteomic approach to ETV5 during endometrial carcinoma invasion reveals a link to oxidative stress. , 2009, Carcinogenesis.

[43]  H. Tsuda,et al.  Clinicopathological and prognostic relevance of uptake level using 18F-fluorodeoxyglucose positron emission tomography/computed tomography fusion imaging (18F-FDG PET/CT) in primary breast cancer. , 2008, Japanese journal of clinical oncology.

[44]  R. Elledge,et al.  Infiltrating lobular carcinoma of the breast: tumor characteristics and clinical outcome , 2004, Breast Cancer Research.

[45]  T. Gansler,et al.  Increased expression of fatty acid synthase (OA-519) in ovarian neoplasms predicts shorter survival. , 1997, Human pathology.

[46]  T. Werner,et al.  Utility of 18 F-FDG PET/CT in pre-surgical risk stratification of patients with breast cancer. , 2019, Hellenic journal of nuclear medicine.

[47]  G. Montgomery,et al.  Fine mapping of variants associated with endometriosis in the WNT4 region on chromosome 1p36. , 2013, International journal of molecular epidemiology and genetics.