Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
• Assymetric Division:
– Self renewal
• Tissue-specific normal stem cells must self-renew throughout
the lifetime of the animal to maintain specific organs
• Cancer stem cells undergo self-renewal to maintain tumor
growth
– Differentiation into phenotypically diverse mature cell
types
• Give rise to a heterogeneous population of cells that
compose the organ or the tumor but lack the ability for
unlimited proliferation (hierarchical arrangement of cells)
• Regulated by similar pathways
– Pathways that regulate self-renewal in normal stem
cells are dys-regulated in cancer stem cells
Development of Hematopoietic Stem Cells
Stem Multipotent Oligolineage Mature
Cells Progenitors Progenitors Cells
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)
Adult intestinal homeostasis
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
Pathways involved in self-renewal that are deregulated in cancer cells
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).
Origin of the Theory of Cancer Stem Cells
Only a small subset of cancer cells is capable of
extensive proliferation
Liquid Tumors
In vitro colony forming assays:
- 1 in 10,000 to 1 in 100 mouse myeloma cells obtained from ascites
away from normal hematopoietic cells were able to form colonies
In vivo transplantation assays:
- Only 1-4% of transplanted leukaemic cells could form spleen
colonies
Solid Tumors
- A large number of cells are required to grow tumors in xenograft
models
- 1 in 1,000 to 1 in 5,000 lung cancer, neuroblastoma cells, ovarian
cancer cells, or breast cancer cells can form colonies in soft agar or
in vivo
Two General Models for Cancer Heterogeneity
• Most therapies fail to consider the difference in drug sensitivities of cancer stem cells
compared to their non-tumorigenic progeny.
• Most therapies target rapidly proliferating non-tumorigenic cells and spare the
relatively quiescent cancer stem cells.
Thymidylate synthase
Raltitrexed 5-FU
Passegue, Emmanuelle et al. (2003) Proc. Natl. Acad. Sci. USA 100, 11842-11849
Self-renewal Assay in NOD/SCID Mice
(Non-obese diabetic/severe combined immunodeficiency)
FACS Cell
Sorter
Cancer Cells
(ex: Leukaemia cells
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)
CD44 Expression
FACS Cell
Sorter
Single Cell
Solid Tumor
Suspension CD24 Expression
Mince Surgical
(small
Implantation
pieces)
Stem Cells in the Nervous System
Brain Tumor Stem Cells: CD133+
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.
CD133+
CD133+ CD133-
Al-Hajj, Muhammad et al. (2003) Proc. Natl. Acad. Sci. USA 100, 3983-3988
FUTURE DIRECTIONS