Isolation and Molecular Characterization of Cancer Stem Cells in MMTV‐Wnt‐1 Murine Breast Tumors

In human breast cancers, a phenotypically distinct minority population of tumorigenic (TG) cancer cells (sometimes referred to as cancer stem cells) drives tumor growth when transplanted into immunodeficient mice. Our objective was to identify a mouse model of breast cancer stem cells that could have relevance to the study of human breast cancer. To do so, we used breast tumors of the mouse mammary tumor virus (MMTV)‐Wnt‐1 mice. MMTV‐Wnt‐1 breast tumors were harvested, dissociated into single‐cell suspensions, and sorted by flow cytometry on Thy1, CD24, and CD45. Sorted cells were then injected into recipient background FVB/NJ female syngeneic mice. In six of seven tumors examined, Thy1+CD24+ cancer cells, which constituted approximately 1%–4% of tumor cells, were highly enriched for cells capable of regenerating new tumors compared with cells of the tumor that did not fit this profile (“not‐Thy1+CD24+”). Resultant tumors had a phenotypic diversity similar to that of the original tumor and behaved in a similar manner when passaged. Microarray analysis comparing Thy1+CD24+ tumor cells to not‐Thy1+CD24+ cells identified a list of differentially expressed genes. Orthologs of these differentially expressed genes predicted survival of human breast cancer patients from two different study groups. These studies suggest that there is a cancer stem cell compartment in the MMTV‐Wnt‐1 murine breast tumor and that there is a clinical utility of this model for the study of cancer stem cells.

[1]  Aleksandar Dakic,et al.  Tumor Growth Need Not Be Driven by Rare Cancer Stem Cells , 2007, Science.

[2]  Michael F. Clarke,et al.  Phenotypic characterization of human colorectal cancer stem cells , 2007, Proceedings of the National Academy of Sciences.

[3]  G. Sherlock,et al.  The prognostic role of a gene signature from tumorigenic breast-cancer cells. , 2007, The New England journal of medicine.

[4]  Stephen C. Harris,et al.  Rat toxicogenomic study reveals analytical consistency across microarray platforms , 2006, Nature Biotechnology.

[5]  T. Golub,et al.  Transformation from committed progenitor to leukaemia stem cell initiated by MLL–AF9 , 2006, Nature.

[6]  D. Fearon,et al.  Identifying genes important for spermatogonial stem cell self-renewal and survival. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Christopher J Ormandy,et al.  Key stages in mammary gland development - The alveolar switch: coordinating the proliferative cues and cell fate decisions that drive the formation of lobuloalveoli from ductal epithelium , 2006, Breast Cancer Research.

[8]  Haiyan I. Li,et al.  Purification and unique properties of mammary epithelial stem cells , 2006, Nature.

[9]  François Vaillant,et al.  Generation of a functional mammary gland from a single stem cell , 2006, Nature.

[10]  L. Holmberg,et al.  Gene expression profiling spares early breast cancer patients from adjuvant therapy: derived and validated in two population-based cohorts , 2005, Breast Cancer Research.

[11]  P. Hall,et al.  An expression signature for p53 status in human breast cancer predicts mutation status, transcriptional effects, and patient survival. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Tao Han,et al.  Cross-platform comparability of microarray technology: Intra-platform consistency and appropriate data analysis procedures are essential , 2005, BMC Bioinformatics.

[13]  T. Jacks,et al.  Identification of Bronchioalveolar Stem Cells in Normal Lung and Lung Cancer , 2005, Cell.

[14]  R. Schiff,et al.  Estrogen receptor positivity in mammary tumors of Wnt-1 transgenic mice is influenced by collaborating oncogenic mutations , 2005, Oncogene.

[15]  G. Glinsky,et al.  Microarray analysis identifies a death-from-cancer signature predicting therapy failure in patients with multiple types of cancer. , 2005, The Journal of clinical investigation.

[16]  I. Kola,et al.  Elf5 is essential for early embryogenesis and mammary gland development during pregnancy and lactation , 2005, The EMBO journal.

[17]  R. Henkelman,et al.  Identification of human brain tumour initiating cells , 2004, Nature.

[18]  S. Aaronson,et al.  An autocrine mechanism for constitutive Wnt pathway activation in human cancer cells. , 2004, Cancer cell.

[19]  S Miyano,et al.  Open source clustering software. , 2004, Bioinformatics.

[20]  Michael F. Clarke,et al.  Applying the principles of stem-cell biology to cancer , 2003, Nature Reviews Cancer.

[21]  C. Boulanger,et al.  Mammary epithelial stem cells: transplantation and self‐renewal analysis , 2003, Cell proliferation.

[22]  I. Weissman,et al.  Expression of BCR/ABL and BCL-2 in myeloid progenitors leads to myeloid leukemias , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[23]  G. Sauvageau,et al.  Bmi-1 determines the proliferative capacity of normal and leukaemic stem cells , 2003, Nature.

[24]  S. Morrison,et al.  Prospective identification of tumorigenic breast cancer cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Irving L. Weissman,et al.  Prospective isolation of human clonogenic common myeloid progenitors , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[26]  T. Ley,et al.  PML/RARα and FLT3-ITD induce an APL-like disease in a mouse model , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[27]  T. Graubert,et al.  Sca-1(pos) cells in the mouse mammary gland represent an enriched progenitor cell population. , 2002, Developmental biology.

[28]  Mina J Bissell,et al.  Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. , 2002, Genes & development.

[29]  T. Ley,et al.  PML/RARalpha and FLT3-ITD induce an APL-like disease in a mouse model. , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Hans Clevers,et al.  The beta-catenin/TCF-4 complex imposes a crypt progenitor phenotype on colorectal cancer cells. , 2002, Cell.

[31]  Van,et al.  A gene-expression signature as a predictor of survival in breast cancer. , 2002, The New England journal of medicine.

[32]  I. Weissman,et al.  Stem cells, cancer, and cancer stem cells , 2001, Nature.

[33]  R. Spang,et al.  Predicting the clinical status of human breast cancer by using gene expression profiles , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[34]  G H Smith,et al.  Mammary epithelial stem cells , 2001, Microscopy research and technique.

[35]  R. Parsons,et al.  APC truncation and increased β-catenin levels in a human breast cancer cell line , 2000 .

[36]  W. Weis,et al.  Structural basis of the Axin–adenomatous polyposis coli interaction , 2000, The EMBO journal.

[37]  R. Parsons,et al.  APC truncation and increased beta-catenin levels in a human breast cancer cell line. , 2000, Carcinogenesis.

[38]  E. Fuchs,et al.  A common human skin tumour is caused by activating mutations in beta-catenin. , 1999, Nature genetics.

[39]  D. Botstein,et al.  Cluster analysis and display of genome-wide expression patterns. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[40]  E. Fuchs,et al.  De Novo Hair Follicle Morphogenesis and Hair Tumors in Mice Expressing a Truncated β-Catenin in Skin , 1998, Cell.

[41]  C. Raffel,et al.  Sporadic Medulloblastomas Contain Oncogenic β-Catenin Mutations , 1998 .

[42]  C. Raffel,et al.  Sporadic medulloblastomas contain oncogenic beta-catenin mutations. , 1998, Cancer research.

[43]  R. Nusse,et al.  Wnt signaling: a common theme in animal development. , 1997, Genes & development.

[44]  J. Dick,et al.  Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell , 1997, Nature Medicine.

[45]  T. Austin,et al.  A role for the Wnt gene family in hematopoiesis: expansion of multilineage progenitor cells. , 1997, Blood.

[46]  D. Gallahan,et al.  The mouse mammary tumor associated gene INT3 is a unique member of the NOTCH gene family (NOTCH4) , 1997, Oncogene.

[47]  Hiromitsu Nakauchi,et al.  Long-Term Lymphohematopoietic Reconstitution by a Single CD34-Low/Negative Hematopoietic Stem Cell , 1996, Science.

[48]  I. Weissman,et al.  The biology of hematopoietic stem cells. , 1995, Annual review of cell and developmental biology.

[49]  M. Caligiuri,et al.  A cell initiating human acute myeloid leukaemia after transplantation into SCID mice , 1994, Nature.

[50]  Bert Vogelstein,et al.  APC mutations occur early during colorectal tumorigenesis , 1992, Nature.

[51]  D. Cox,et al.  Mode of proviral activation of a putative mammary oncogene (int-1) on mouse chromosome 15 , 1984, Nature.