Cancer Stem Cells. Maria M. (Marj) Pe ña, PhD Dept. of Biological Sciences Center for Colon Cancer Research University of South Carolina. Normal stem cells. Rare cells within organs with the ability to self-renew and give rise to all types of cells within the organ to drive organogenesis.
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Maria M. (Marj) Peña, PhD
Dept. of Biological Sciences
Center for Colon Cancer Research
University of South Carolina
Rare cells within organs with the ability to self-renew and give rise to all types of cells within the organ to drive organogenesis
Cancer stem cells
Rare cells within tumors with the ability to self-renew and give rise to the phenotypically diverse tumor cell population to drive tumorigenesis
HSCs can be subdivided into long-term self-renewing HSCs, short-term self-renewing HSCs and multipotent progenitors (red arrows indicate self-renewal). They give rise to common lymphoid progenitors (CLPs; the precursors of all lymphoid cells) and common myeloid progenitors (CMPs; the precursors of all myeloid cells). Both CMPs/GMPs (granulocyte macrophage precursors) and CLPs can give rise to all known mouse dendritic cells. ErP, erythrocyte precursor; MEP, megakaryocyte erythrocyte precursor; MkP, megakaryocyte precursor; NK, natural killer.
Reya et al. 2001 Nature 414:105-111
The epithelium is shaped into crypts and villi (left). The lineage scheme (right) depicts the stem cell, the transit-amplifying cells, and the two differentiated branches. The right branch constitutes the enterocyte lineage; the left is the secretory lineage. Relative positions along the crypt-villus axis correspond to the schematic graph of the crypt in the center.
F. Radtke et al., Science 307, 1904 -1909 (2005)
Schematic representation and section of the crypt-villus unit in the mature small intestine. Proliferative cells reside in the crypts, while differentiated cells occupy the villus. Crypt progenitors migrate up (red arrow) the crypt-villus axis before shedding into the lumen. The process of epithelial renewal takes 3-6 d and is ensured by a small number of asymmetrically dividing stem cells at the bottom of the crypts. Wnt signaling in the adult intestine promotes proliferation of progenitor or transit-amplifying (TA) cells, as well as commitment toward secretory lineages. Wnt signaling may also drive terminal differentiation of certain secretory lineages. Although it is commonly believed that Wnt signaling may promote proliferation and/or differentiation of intestinal stem cells, there is no evidence that formally proves this (see arrows with question marks). In panel A, black arrowheads indicate Ki67 positive transit-amplifying cells, while white arrowheads indicate the Paneth cell compartment.
Alex Gregorieff et al. Genes Dev. 2005; 19: 877-890
Wnt, Shh, and Notch pathways have been shown to contribute to the self-renewal of stem cells and/or progenitors in a variety of organs, including the haematopoietic and nervous systems. When dysregulated, these pathways can contribute to oncogenesis. Mutations of these pathways have been associated with a number of human tumours, including colon carcinoma and epidermal tumours (Wnt), medulloblastoma and basal cell carcinoma (Shh), and T-cell leukaemias (Notch).
1. All cancer cells are potential cancer stem cells but have a low probability of proliferation in clonogenic assays
2. Only a small definable subset of cancer cells are cancer stem cells that have the ability to proliferate indefinitely.
All cells have equal but low probability of
extensive proliferation. Only cells with self
renewal capacity can sustain tumor growth.
Distinct classes of cells exist within a
tumor. Only a small definable subset,
the cancer stem cells can initiate tumor
Chu E. et al., Cancer Chemother Pharmacol (2003) 52 (Suppl 1) S80-S89
Longley, DB et al., Nature Reviews Cancer (2003) 3:330-338
Passegue, Emmanuelle et al. (2003) Proc. Natl. Acad. Sci. USA 100, 11842-11849
(Non-obese diabetic/severe combined immunodeficiency)
(ex: Leukaemia cells
Sublethally irradiated NOD/SCID Mice
Human hematopoietic cells are organized in a hierarchy that is sustained by a small population of self-renewing hematopoietic stem cells (HSCs). HSCs give rise to progressively more lineage-restricted, differentiated progenitors with reduced self-renewal capacity (LTC-ICs, long-term culture-initiating cells; CFU, colony-forming units), which in turn produce functionally mature blood cells.
Disruption of pathways regulating self-renewal and differentiation through the acquisition of transforming mutations generates leukemic stem cells (LSCs) capable of sustaining growth of the leukemic clone in vivo. LSCs possess an altered differentiation program, as demonstrated by aberrant expression of some cell-surface markers (indicated in blue) and give rise to an aberrant developmental hierarchy that retains aspects of its normal counterpart.
In vivo reconstitution assays using immune-deficient mouse recipients enable detection of HSCs and LSCs as SCID-repopulating cells (SRCs) and SCID leukemia-initiating cells (SL-ICs), respectively.
Wang and Dick 2005 Trends in Cell Biology 15:494-501
Acute myeloid leukemia
(AML) –CD34+ CD38-
Leukaemic Mouse Models:
chronic myelomonocytic leukaemia (CMML) MRP8-BCL-2
acute myeloid leukaemia (AML)MRP8-BCL2Xlpr/lpr
chronic myeloid leukaemia (CML)/BlastMRP8-PML-RARα
acute promyelocytic leukaemia (APML)77MRP8-BCRablXBCL-2
Self-renewal is a key property of both normal and leukemic stem cells. Fewer mutagenic changes are required to transform stem cells in which the self-renewal machinery is already active (a), as compared with committed progenitors in which self-renewal must be activated ectopically (b). In addition, self-renewing stem cells are long-lived; thus, there is an increased chance for genetic changes to accumulate in individual stem cells in comparison with more mature, short-lived progenitors. If a committed progenitor with limited life span acquires a genetic mutation that does not confer increased self-renewal (c), that cell will likely die or undergo terminal differentiation before enough mutations occur to propagate a full leukemogenic program.
Self-renewal Assay in NOD/SCID Mice
For solid tumors: surgical orthotopic implantation (SOI)
CD133 – neuronal stem cell marker
Brain tumor stem cells were identified from human brain tumor samples by in vitro neurosphere assays normally used to isolate normal neural stem cells
GFAP = glial fibrillary acidic protein
Singh et. al 2003 Cancer Research 63: 5821-5828.
Brain tumor stem cells were identified by intracranial transplantation of CD133+ cells into adult NOD/SCID mouse forebrain.
Singh et al. 2004 Nature 432: 396-401
CD44 and CD24 – adhesion molecules
B38.1 – breast/ovarian cancer-specific marker
ESA – epithelial specific antigen
Al-Hajj, Muhammad et al. (2003) Proc. Natl. Acad. Sci. USA 100, 3983-3988
Vascularization of primary tumor
Tumor grows through the synthesis and secretion of pro-angiogenic factors by the tumor and surrounding stroma
Invasion of the organ stroma through enhanced expression of enzymes (MMP9)
Invasion of the lymphatic or vascular channels (may grow in these places)
Tumors cells enter circulation
Must survive turbulence of circulation and evade both immune and non-immune mechanisms
Tumors are biologically heterogeneous and contain subpopulations of cells with different angiogenic, invasive, and metastatic properties.
MDA-MB-231 Breast Cancer Cell Line
Isolate Single Clonal Populations (SCPs)
Introduce Luciferase Bioluminescent Marker and GFP Fluorescence Marker
Introduce into Nude Mice by intracardiac Injection
Minn, A. J. et al. J. Clin. Invest. 2005;115:44-55
Noninvasive BLI to monitor the development of osteolytic metastases from the same mouse
Verification of macroscopic and microscopic metastases by fluorescence histology
to metastasize to the lung
Metastases is a selective process for cells that succeed in invasion, embolization, survival in the circulation, arrest in a distal capillary bed, extravastion into the distant organ, and survival and proliferation in the distant organ.
SCPs from MDA-MB-231 cells have a poor-prognosis gene expression signature
Minn, A. J. et al. J. Clin. Invest. 2005;115:44-55
CXCR4 – bone homing chemokine receptor
CTGF – connective tissue growth factor
IL-11 – activator of osteoclast differentiation (mediators of bone resorption in bone metastases)
MMP1 – matrix metalloproteinase/collagenase, promotes osteolysis by cleaving
a specific peptide bond in the collagen of bone matrix
OPN – osteopontin (consistently overexpressed in metastatic cells)
The outcome of metastasis depends on multiple interactions (“cross-talk”) between the metastatic subpopulation in the primary tumor and the host organ microenvironment.
Fodde, R et al., Nat Rev Cancer (2001) 1:57-67