Propagated circulating tumor cells uncovers the role of NFκB and COP1 in metastasis

Circulating tumor cells (CTCs), a population of cancer cells that represents the seeds of metastatic nodules, are a promising model system for studying metastasis. However, expansion of patient-derived CTCs ex vivo is challenging and dependent on the collection of high numbers of CTCs, which are ultra-rare. Here, we report the development of a combined CTC and CTC-derived xenograft (CDX) platform for expanding and studying patient-derived CTCs from metastatic colon, lung, and pancreatic cancers. Propagated CTCs yielded a highly aggressive population of cells that could be used to routinely and robustly establish primary tumors and metastatic lesions in CDXs. Differential gene analysis of the resultant CTC models emphasized a role for NF-kB signaling as a pan-cancer signaling pathway involved in metastasis. Furthermore, metastatic CTCs were identified through a prospective 5-gene signature (BCAR1, COL1A1, IGSF3, RRAD, and TFPI2). Whole-exome sequencing of CDX models and metastases further identified mutations in constitutive photomorphogenesis protein 1 (COP1) as a potential driver of metastasis. These findings illustrate the utility of the combined patient-derived CTC model and provide a glimpse of the promise of CTCs in identifying drivers of cancer metastasis.

[1]  Jia Fan,et al.  Single-cell RNA sequencing reveals spatial heterogeneity and immune evasion of circulating tumor cells , 2021, Cancer biology & medicine.

[2]  C. Isaacs,et al.  Circulating Tumor Cells: Technologies and Their Clinical Potential in Cancer Metastasis , 2021, Biomedicines.

[3]  Xiaochen Bo,et al.  clusterProfiler 4.0: A universal enrichment tool for interpreting omics data , 2021, Innovation.

[4]  C. Lindskog,et al.  A single–cell type transcriptomics map of human tissues , 2021, Science Advances.

[5]  David R. Jones,et al.  Genomic characterization of metastatic patterns from prospective clinical sequencing of 25,000 patients , 2021, Cell.

[6]  W. Wang,et al.  TFPI2 Promotes Perivascular Migration in an Angiotropism Model of Melanoma , 2021, Frontiers in Oncology.

[7]  J. Cueva,et al.  Short-Term Ex Vivo Culture of CTCs from Advance Breast Cancer Patients: Clinical Implications , 2021, Cancers.

[8]  K. Itahana,et al.  Mutant TP53 interacts with BCAR1 to contribute to cancer cell invasion , 2020, British journal of cancer.

[9]  Yanyan Zhang,et al.  NUF2 as an anticancer therapeutic target and prognostic factor in breast cancer. , 2020, International journal of oncology.

[10]  S. Jeffrey,et al.  Detection of EGFR Mutations in cfDNA and CTCs, and Comparison to Tumor Tissue in Non-Small-Cell-Lung-Cancer (NSCLC) Patients , 2020, Frontiers in Oncology.

[11]  Kelly P. Stanton,et al.  Efficient Propagation of Circulating Tumor Cells: A First Step for Probing Tumor Metastasis , 2020, Cancers.

[12]  T. Tan,et al.  Characterization of circulating breast cancer cells with tumorigenic and metastatic capacity , 2020, EMBO molecular medicine.

[13]  S. Cairo,et al.  Genetic characterization of a unique neuroendocrine transdifferentiation prostate circulating tumor cell-derived eXplant model , 2020, Nature Communications.

[14]  T. Lawrence,et al.  Expansion of Circulating Tumor Cells from Patients with Locally Advanced Pancreatic Cancer Enable Patient Derived Xenografts and Functional Studies for Personalized Medicine , 2020, Cancers.

[15]  Hongchi Jiang,et al.  The immunoglobulin superfamily member 3 (IGSF3) promotes hepatocellular carcinoma progression through activation of the NF-κB pathway , 2020, Annals of translational medicine.

[16]  M. Carolan,et al.  Establishment of novel long-term cultures from EpCAM positive and negative circulating tumour cells from patients with metastatic gastroesophageal cancer , 2020, Scientific Reports.

[17]  Joon-Oh Park,et al.  RRAD expression in gastric and colorectal cancer with peritoneal carcinomatosis , 2019, Scientific Reports.

[18]  M. Jücker,et al.  Distinct functions of AKT isoforms in breast cancer: a comprehensive review , 2019, Cell Communication and Signaling.

[19]  Gennady Korotkevich,et al.  Fast gene set enrichment analysis , 2019, bioRxiv.

[20]  Atul Padole,et al.  Diffuse Lung Metastases in EGFR-Mutant Non-Small Cell Lung Cancer , 2019, Cancers.

[21]  O. Straume,et al.  Are 90% of deaths from cancer caused by metastases? , 2019, Cancer medicine.

[22]  Wei-Hwa Lee,et al.  Collagen 1A1 (COL1A1) Is a Reliable Biomarker and Putative Therapeutic Target for Hepatocellular Carcinogenesis and Metastasis , 2019, Cancers.

[23]  M. Cristofanilli,et al.  Lymphomagenesis in Circulating Tumor Cell-Derived Xenograft Models , 2019, Biomedical Journal of Scientific & Technical Research.

[24]  Ash A. Alizadeh,et al.  Determining cell-type abundance and expression from bulk tissues with digital cytometry , 2019, Nature Biotechnology.

[25]  S. Volinia,et al.  Heterogeneity in Circulating Tumor Cells: The Relevance of the Stem-Cell Subset , 2019, Cancers.

[26]  I. Roninson,et al.  Role of transcription-regulating kinase CDK8 in colon cancer metastasis , 2019, Oncotarget.

[27]  I. Abdulkader,et al.  CTCs‐derived xenograft development in a triple negative breast cancer case , 2018, International journal of cancer.

[28]  Jacqueline A. Hall,et al.  A framework for the development of effective anti-metastatic agents , 2018, Nature Reviews Clinical Oncology.

[29]  W. Gu,et al.  TFPI-2 suppresses breast cancer cell proliferation and invasion through regulation of ERK signaling and interaction with actinin-4 and myosin-9 , 2018, Scientific Reports.

[30]  Ben S. Wittner,et al.  A Digital RNA Signature of Circulating Tumor Cells Predicting Early Therapeutic Response in Localized and Metastatic Breast Cancer. , 2018, Cancer discovery.

[31]  Yong Teng,et al.  Combating head and neck cancer metastases by targeting Src using multifunctional nanoparticle-based saracatinib , 2018, Journal of Hematology & Oncology.

[32]  Mehmet Toner,et al.  Genomic and Functional Fidelity of Small Cell Lung Cancer Patient-Derived Xenografts. , 2018, Cancer discovery.

[33]  Jinghang Zhang,et al.  COL1A1 promotes metastasis in colorectal cancer by regulating the WNT/PCP pathway , 2018, Molecular medicine reports.

[34]  M. Pitman,et al.  Author Correction: Pancreatic cancer: Circulating Tumor Cells and Primary Tumors show Heterogeneous KRAS Mutations , 2017, Scientific Reports.

[35]  C. Dive,et al.  Circulating tumor cells and CDX models as a tool for preclinical drug development. , 2017, Translational lung cancer research.

[36]  D. Weitz,et al.  An RNA-based signature enables high specificity detection of circulating tumor cells in hepatocellular carcinoma , 2017, Proceedings of the National Academy of Sciences.

[37]  De-sheng Wang,et al.  RRAD inhibits aerobic glycolysis, invasion, and migration and is associated with poor prognosis in hepatocellular carcinoma , 2016, Tumor Biology.

[38]  P. Steeg,et al.  Targeting metastasis , 2016, Nature Reviews Cancer.

[39]  C. Dive,et al.  Tumourigenic non-small-cell lung cancer mesenchymal circulating tumour cells: a clinical case study , 2016, Annals of oncology : official journal of the European Society for Medical Oncology.

[40]  N. Dhomen,et al.  Application of Sequencing, Liquid Biopsies, and Patient-Derived Xenografts for Personalized Medicine in Melanoma. , 2016, Cancer discovery.

[41]  P. O’Donnell,et al.  Saracatinib as a metastasis inhibitor in metastatic castration‐resistant prostate cancer: A University of Chicago Phase 2 Consortium and DOD/PCF Prostate Cancer Clinical Trials Consortium Study , 2016, The Prostate.

[42]  C. Sonnenschein,et al.  Cancer Metastases: So Close and So Far. , 2015, Journal of the National Cancer Institute.

[43]  M. Gazouli,et al.  Mutational Analysis of Circulating Tumor Cells from Colorectal Cancer Patients and Correlation with Primary Tumor Tissue , 2015, PloS one.

[44]  Theresa Zhang,et al.  Personalized genomic analyses for cancer mutation discovery and interpretation , 2015, Science Translational Medicine.

[45]  Udo Schumacher,et al.  Establishment and characterization of a cell line from human circulating colon cancer cells. , 2015, Cancer research.

[46]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[47]  Hans Clevers,et al.  Organoid Cultures Derived from Patients with Advanced Prostate Cancer , 2014, Cell.

[48]  Sridhar Ramaswamy,et al.  Circulating Tumor Cell Clusters Are Oligoclonal Precursors of Breast Cancer Metastasis , 2014, Cell.

[49]  Sridhar Ramaswamy,et al.  Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility , 2014, Science.

[50]  S. Jeffrey,et al.  Single cell mutational analysis of PIK3CA in circulating tumor cells and metastases in breast cancer reveals heterogeneity, discordance, and mutation persistence in cultured disseminated tumor cells from bone marrow , 2014, BMC Cancer.

[51]  Mahmood Ayub,et al.  Tumorigenicity and genetic profiling of circulating tumor cells in small-cell lung cancer , 2014, Nature Medicine.

[52]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[53]  M. Rugge,et al.  Retaining the long-survive capacity of Circulating Tumor Cells (CTCs) followed by xeno-transplantation: not only from metastatic cancer of the breast but also of prostate cancer patients , 2013, Oncoscience.

[54]  Jiahuai Han,et al.  COP1 and GSK3β cooperate to promote c-Jun degradation and inhibit breast cancer cell tumorigenesis. , 2013, Neoplasia.

[55]  Peter Ulz,et al.  Complex tumor genomes inferred from single circulating tumor cells by array-CGH and next-generation sequencing. , 2013, Cancer research.

[56]  Tim Holland-Letz,et al.  Identification of a population of blood circulating tumor cells from breast cancer patients that initiates metastasis in a xenograft assay , 2013, Nature Biotechnology.

[57]  Wei Yin,et al.  The Identification and Characterization of Breast Cancer CTCs Competent for Brain Metastasis , 2013, Science Translational Medicine.

[58]  K. Kinzler,et al.  Cancer Genome Landscapes , 2013, Science.

[59]  Sridhar Ramaswamy,et al.  Circulating Breast Tumor Cells Exhibit Dynamic Changes in Epithelial and Mesenchymal Composition , 2013, Science.

[60]  C. V. Jongeneel,et al.  Exome sequencing identifies recurrent somatic MAP2K1 and MAP2K2 mutations in melanoma , 2011, Nature Genetics.

[61]  N. Sethi,et al.  Notch signalling in cancer progression and bone metastasis , 2011, British Journal of Cancer.

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

[63]  K. Isselbacher,et al.  Isolation of circulating tumor cells using a microvortex-generating herringbone-chip , 2010, Proceedings of the National Academy of Sciences.

[64]  W. Gerald,et al.  Genes that mediate breast cancer metastasis to the brain , 2009, Nature.

[65]  Cole Trapnell,et al.  Ultrafast and memory-efficient alignment of short DNA sequences to the human genome , 2009, Genome Biology.

[66]  K. Pienta,et al.  Circulating Tumor Cells Predict Survival Benefit from Treatment in Metastatic Castration-Resistant Prostate Cancer , 2008, Clinical Cancer Research.

[67]  Mehmet Toner,et al.  Detection of mutations in EGFR in circulating lung-cancer cells. , 2008, The New England journal of medicine.

[68]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[69]  Andy J. Minn,et al.  Genes that mediate breast cancer metastasis to lung , 2005, Nature.

[70]  Xavier Matias-Guiu,et al.  NF-kB in development and progression of human cancer , 2005, Virchows Archiv.

[71]  I. Fidler,et al.  Metastasis results from preexisting variant cells within a malignant tumor. , 1977, Science.

[72]  J. Lorente,et al.  Circulating Tumor Cells: Markers and Methodologies for Enrichment and Detection. , 2017, Methods in molecular biology.

[73]  J. Mesirov,et al.  The Molecular Signatures Database (MSigDB) hallmark gene set collection. , 2015, Cell systems.

[74]  T. Seyfried,et al.  On the origin of cancer metastasis. , 2013, Critical reviews in oncogenesis.

[75]  Emmanuel A Theodorakis,et al.  Anti-TNF-alpha therapies: the next generation. , 2003, Nature reviews. Drug discovery.

[76]  Hiroyuki Ogata,et al.  KEGG: Kyoto Encyclopedia of Genes and Genomes , 1999, Nucleic Acids Res..