Targeting stromal remodeling and cancer stem cell plasticity to overcome chemoresistance in triple negative breast cancer

The cellular and molecular basis of stromal cell recruitment, activation and crosstalk in carcinomas is poorly understood, limiting the development of targeted anti-stromal therapies. In mouse models of triple negative breast cancer (TNBC), Hh ligand produced by neoplastic cells reprogrammed cancer-associated fibroblast (CAF) gene expression, driving tumor growth and metastasis. Hh-activated CAFs upregulated expression of FGF5 and production of fibrillar collagen, leading to FGFR and FAK activation in adjacent neoplastic cells, which then acquired a stem-like, drug-resistant phenotype. Treatment with smoothened inhibitors (SMOi) reversed these phenotypes. Stromal treatment of TNBC patient-derived xenograft (PDX) models with SMOi downregulated the expression of cancer stem cell markers and sensitized tumors to docetaxel, leading to markedly improved survival and reduced metastatic burden. In the phase I clinical trial EDALINE, 3 of 12 patients with metastatic TNBC derived clinical benefit from combination therapy with the SMOi Sonidegib and docetaxel chemotherapy, with one patient experiencing a complete response. Markers of pathway activity correlated with response. These studies identify Hh signaling to CAFs as a novel mediator of cancer stem cell plasticity and an exciting new therapeutic target in TNBC. SIGNIFICANCE Compared to other breast cancer subtypes, TNBCs are associated with significantly worse patient outcomes. Standard of care systemic treatment for patients with non-BRCA1/2 positive TNBC is cytotoxic chemotherapy. However, the failure of 70% of treated TNBCs to attain complete pathological response reflects the relative chemoresistance of these tumors. New therapeutic strategies are needed to improve patient survival and quality of life. Here, we provide new insights into the dynamic interactions between heterotypic cells within a tumor. Specifically, we establish the mechanisms by which CAFs define cancer cell phenotype and demonstrate that the bidirectional CAF-cancer cell crosstalk can be successfully targeted in mice and humans using anti-stromal therapy.

[1]  J. Erler,et al.  ISDoT: in situ decellularization of tissues for high-resolution imaging and proteomic analysis of native extracellular matrix , 2017, Nature Medicine.

[2]  W. Birchmeier,et al.  Cancer Stem Cells Regulate Cancer-Associated Fibroblasts via Activation of Hedgehog Signaling in Mammary Gland Tumors. , 2017, Cancer research.

[3]  Max Nobis,et al.  Transient tissue priming via ROCK inhibition uncouples pancreatic cancer progression, sensitivity to chemotherapy, and metastasis , 2017, Science Translational Medicine.

[4]  N. Turner,et al.  Advances and challenges in targeting FGFR signalling in cancer , 2017, Nature Reviews Cancer.

[5]  D. Tuveson,et al.  Br Ief Definitive Repor T , 2022 .

[6]  C. Bult,et al.  Patient-derived xenograft (PDX) models in basic and translational breast cancer research , 2016, Cancer and Metastasis Reviews.

[7]  D. Radisky,et al.  Tissue Stiffness and Hypoxia Modulate the Integrin-Linked Kinase ILK to Control Breast Cancer Stem-like Cells. , 2016, Cancer research.

[8]  S. Joost,et al.  Stromal Hedgehog signalling is downregulated in colon cancer and its restoration restrains tumour growth , 2016, Nature Communications.

[9]  V. Weaver,et al.  Force Matters: Biomechanical Regulation of Cell Invasion and Migration in Disease. , 2016, Trends in cell biology.

[10]  I. Ng,et al.  Cancer-Associated Fibroblasts Regulate Tumor-Initiating Cell Plasticity in Hepatocellular Carcinoma through c-Met/FRA1/HEY1 Signaling. , 2016, Cell reports.

[11]  C. Streuli,et al.  Raised mammographic density: causative mechanisms and biological consequences , 2016, Breast Cancer Research.

[12]  Christine A Iacobuzio-Donahue,et al.  Genotype tunes pancreatic ductal adenocarcinoma tissue tension to induce matricellular-fibrosis and tumor progression , 2016, Nature Medicine.

[13]  D. V. Hoff,et al.  Hedgehog Pathway Inhibition , 2016, Cell.

[14]  B. Maček,et al.  Differential Rac1 signalling by guanine nucleotide exchange factors implicates FLII in regulating Rac1-driven cell migration , 2016, Nature Communications.

[15]  Angela E. Leek,et al.  Increased peri-ductal collagen micro-organization may contribute to raised mammographic density , 2016, Breast Cancer Research.

[16]  Natasha Kolesnikoff,et al.  A Negative Regulatory Mechanism Involving 14-3-3ζ Limits Signaling Downstream of ROCK to Regulate Tissue Stiffness in Epidermal Homeostasis. , 2015, Developmental cell.

[17]  G. Wahl,et al.  Sox10 Regulates Stem/Progenitor and Mesenchymal Cell States in Mammary Epithelial Cells. , 2015, Cell reports.

[18]  Chih-Yang Wang,et al.  Single-cell analysis reveals a stem-cell program in human metastatic breast cancer cells , 2015, Nature.

[19]  Astrid Magenau,et al.  High mammographic density is associated with an increase in stromal collagen and immune cells within the mammary epithelium , 2015, Breast Cancer Research.

[20]  Evan Z. Macosko,et al.  Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets , 2015, Cell.

[21]  F. Sotgia,et al.  Chemotherapy induces the cancer-associated fibroblast phenotype, activating paracrine Hedgehog-GLI signalling in breast cancer cells , 2015, Oncotarget.

[22]  Erik Sahai,et al.  Intravital Imaging Reveals How BRAF Inhibition Generates Drug-Tolerant Microenvironments with High Integrin β1/FAK Signaling , 2015, Cancer cell.

[23]  J. McCaffrey,et al.  The fate of chemoresistance in triple negative breast cancer (TNBC) , 2015, BBA clinical.

[24]  D. Sahoo,et al.  Hedgehog signaling restrains bladder cancer progression by eliciting stromal production of urothelial differentiation factors. , 2014, Cancer cell.

[25]  S. Ran,et al.  Paclitaxel therapy promotes breast cancer metastasis in a TLR4-dependent manner. , 2014, Cancer research.

[26]  C. Streuli,et al.  FGF ligands of the postnatal mammary stroma regulate distinct aspects of epithelial morphogenesis , 2014, Development.

[27]  S. Dudoit,et al.  Normalization of RNA-seq data using factor analysis of control genes or samples , 2014, Nature Biotechnology.

[28]  J. Willmann,et al.  Stromal response to Hedgehog signaling restrains pancreatic cancer progression , 2014, Proceedings of the National Academy of Sciences.

[29]  Stephen A. Sastra,et al.  Stromal elements act to restrain, rather than support, pancreatic ductal adenocarcinoma. , 2014, Cancer cell.

[30]  Ker-Chau Li,et al.  Cancer-associated fibroblasts regulate the plasticity of lung cancer stemness via paracrine signalling , 2014, Nature Communications.

[31]  S. Araújo,et al.  Hedgehog Is a Positive Regulator of FGF Signalling during Embryonic Tracheal Cell Migration , 2014, PloS one.

[32]  A. Iavarone,et al.  The ID proteins: master regulators of cancer stem cells and tumour aggressiveness , 2014, Nature Reviews Cancer.

[33]  K. Sukhdeo,et al.  Chemotherapy activates cancer-associated fibroblasts to maintain colorectal cancer-initiating cells by IL-17A , 2013, The Journal of experimental medicine.

[34]  M. Frydenberg,et al.  Hedgehog signaling is active in human prostate cancer stroma and regulates proliferation and differentiation of adjacent epithelium , 2013, The Prostate.

[35]  Zainab Jagani,et al.  Unraveling the therapeutic potential of the Hedgehog pathway in cancer , 2013, Nature Medicine.

[36]  Corbin E. Meacham,et al.  Tumour heterogeneity and cancer cell plasticity , 2013, Nature.

[37]  M. Olson,et al.  Mechanotransduction pathways promoting tumor progression are activated in invasive human squamous cell carcinoma. , 2013, The American journal of pathology.

[38]  Robert A. Weinberg,et al.  Poised Chromatin at the ZEB1 Promoter Enables Breast Cancer Cell Plasticity and Enhances Tumorigenicity , 2013, Cell.

[39]  Benjamin E. Gross,et al.  Integrative Analysis of Complex Cancer Genomics and Clinical Profiles Using the cBioPortal , 2013, Science Signaling.

[40]  Sandra A O'Toole,et al.  The Hedgehog signalling pathway in breast development, carcinogenesis and cancer therapy , 2013, Breast Cancer Research.

[41]  Joseph M. Negri,et al.  The role of tumour–stromal interactions in modifying drug response: challenges and opportunities , 2013, Nature Reviews Drug Discovery.

[42]  S. Hilsenbeck,et al.  Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function , 2013, Stem cells.

[43]  F. Rösel,et al.  Mammary fibroblasts regulate morphogenesis of normal and tumorigenic breast epithelial cells by mechanical and paracrine signals. , 2012, Cancer letters.

[44]  T. Wilgus,et al.  Growth Factor-Extracellular Matrix Interactions Regulate Wound Repair. , 2012, Advances in wound care.

[45]  Norbert Perrimon,et al.  Signaling mechanisms controlling cell fate and embryonic patterning. , 2012, Cold Spring Harbor perspectives in biology.

[46]  Steven J. M. Jones,et al.  Comprehensive molecular portraits of human breast tumors , 2012, Nature.

[47]  Jane Fridlyand,et al.  Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors , 2012, Nature.

[48]  T. Golub,et al.  Tumor microenvironment induces innate RAF-inhibitor resistance through HGF secretion , 2012, Nature.

[49]  Wenjun Guo,et al.  Slug and Sox9 Cooperatively Determine the Mammary Stem Cell State , 2012, Cell.

[50]  J. Schneider,et al.  Hedgehog signaling inhibition blocks growth of resistant tumors through effects on tumor microenvironment. , 2012, Cancer research.

[51]  G. Wahl,et al.  A mammary stem cell population identified and characterized in late embryogenesis reveals similarities to human breast cancer. , 2012, Cell stem cell.

[52]  Yue Meng,et al.  The enhancement of cancer stem cell properties of MCF-7 cells in 3D collagen scaffolds for modeling of cancer and anti-cancer drugs. , 2012, Biomaterials.

[53]  Hong Peng,et al.  Interactions between cancer stem cells and their niche govern metastatic colonization , 2011, Nature.

[54]  Mark T. W. Ebbert,et al.  Tumor grafts derived from women with breast cancer authentically reflect tumor pathology, growth, metastasis and disease outcomes , 2011, Nature Medicine.

[55]  P. Timpson,et al.  Actomyosin-mediated cellular tension drives increased tissue stiffness and β-catenin activation to induce epidermal hyperplasia and tumor growth. , 2011, Cancer cell.

[56]  P. Schofield,et al.  Hedgehog overexpression is associated with stromal interactions and predicts for poor outcome in breast cancer. , 2011, Cancer research.

[57]  Helga Thorvaldsdóttir,et al.  Molecular signatures database (MSigDB) 3.0 , 2011, Bioinform..

[58]  S. Hilsenbeck,et al.  Keratin 6a marks mammary bipotential progenitor cells that can give rise to a unique tumor model resembling human normal-like breast cancer , 2011, Oncogene.

[59]  D. Hanahan,et al.  Hallmarks of Cancer: The Next Generation , 2011, Cell.

[60]  E. Lander,et al.  Estrogen expands breast cancer stem-like cells through paracrine FGF/Tbx3 signaling , 2010, Proceedings of the National Academy of Sciences.

[61]  T. Yau,et al.  A subpopulation of CD26+ cancer stem cells with metastatic capacity in human colorectal cancer. , 2010, Cell stem cell.

[62]  Mikala Egeblad,et al.  Matrix Crosslinking Forces Tumor Progression by Enhancing Integrin Signaling , 2009, Cell.

[63]  Davis J. McCarthy,et al.  edgeR: a Bioconductor package for differential expression analysis of digital gene expression data , 2009, Bioinform..

[64]  Riccardo Cicchi,et al.  Scoring of collagen organization in healthy and diseased human dermis by multiphoton microscopy , 2009, Journal of biophotonics.

[65]  W. Bushman,et al.  The Sonic Hedgehog Pathway Stimulates Prostate Tumor Growth by Paracrine Signaling and Recaptures Embryonic Gene Expression in Tumor Myofibroblasts , 2009, Oncogene.

[66]  G. Smyth,et al.  ELDA: extreme limiting dilution analysis for comparing depleted and enriched populations in stem cell and other assays. , 2009, Journal of immunological methods.

[67]  S. Fox,et al.  Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers , 2009, Nature Medicine.

[68]  J. Visvader,et al.  The mammary progenitor marker CD61/beta3 integrin identifies cancer stem cells in mouse models of mammary tumorigenesis. , 2008, Cancer research.

[69]  J. Visvader,et al.  Cancer stem cells in solid tumours: accumulating evidence and unresolved questions , 2008, Nature Reviews Cancer.

[70]  Z. Werb,et al.  Genetic mosaic analysis reveals FGF receptor 2 function in terminal end buds during mammary gland branching morphogenesis. , 2008, Developmental biology.

[71]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[72]  Wenjun Guo,et al.  The Epithelial-Mesenchymal Transition Generates Cells with Properties of Stem Cells , 2008, Cell.

[73]  Susan G Hilsenbeck,et al.  Intrinsic resistance of tumorigenic breast cancer cells to chemotherapy. , 2008, Journal of the National Cancer Institute.

[74]  Hua Tian,et al.  A paracrine requirement for hedgehog signalling in cancer , 2008, Nature.

[75]  F. Pépin,et al.  Stromal gene expression predicts clinical outcome in breast cancer , 2008, Nature Medicine.

[76]  J. Lieberman,et al.  let-7 Regulates Self Renewal and Tumorigenicity of Breast Cancer Cells , 2007, Cell.

[77]  C. Heeschen,et al.  Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. , 2007, Cell stem cell.

[78]  Z. Werb,et al.  Candidate regulators of mammary branching morphogenesis identified by genome‐wide transcript analysis , 2006, Developmental dynamics : an official publication of the American Association of Anatomists.

[79]  J. Mesirov,et al.  From the Cover: Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005 .

[80]  M. Scott,et al.  Incredible journey: how do developmental signals travel through tissue? , 2004, Genes & development.

[81]  A. McMahon,et al.  Feedback control of mammalian Hedgehog signaling by the Hedgehog-binding protein, Hip1, modulates Fgf signaling during branching morphogenesis of the lung. , 2003, Genes & development.

[82]  Zena Werb,et al.  Stromal Effects on Mammary Gland Development and Breast Cancer , 2002, Science.

[83]  C. Sugnet,et al.  The Gli2 transcription factor is required for normal mouse mammary gland development. , 2001, Developmental biology.

[84]  C. Murre,et al.  Regulation of the helix-loop-helix proteins, E2A and Id3, by the Ras-ERK MAPK cascade , 2001, Nature Immunology.

[85]  Abdul Kader Sagar,et al.  Second-harmonic generation imaging of cancer. , 2014, Methods in cell biology.

[86]  Steven J. M. Jones,et al.  Comprehensive molecular portraits of human breast tumours , 2013 .

[87]  P. Keely,et al.  Bi-directional signaling: extracellular matrix and integrin regulation of breast tumor progression. , 2013, Critical reviews in eukaryotic gene expression.

[88]  Alexandra Naba,et al.  Overview of the matrisome--an inventory of extracellular matrix constituents and functions. , 2012, Cold Spring Harbor perspectives in biology.

[89]  Tzong-Shiue Yu,et al.  A restricted cell population propagates glioblastoma growth after chemotherapy , 2012 .

[90]  L. Schwartz,et al.  New response evaluation criteria in solid tumours: revised RECIST guideline (version 1.1). , 2009, European journal of cancer.

[91]  David Cameron,et al.  A stroma-related gene signature predicts resistance to neoadjuvant chemotherapy in breast cancer , 2009, Nature Medicine.

[92]  Jeffrey E. Green,et al.  Development and Characterization of a Progressive Series of Mammary Adenocarcinoma Cell Lines Derived from the C3(1)/SV40 Large T-antigen Transgenic Mouse Model , 2004, Breast Cancer Research and Treatment.