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Cancer stem cells:  implications in the  clinical setting  Malcolm Alison; m.alison@qmul.ac.uk 4 th  April 2011
 
 
 
Time Initiator Promoter Initiators  &   promoters * Tumours Tumours No tumours DMBA TPA
 
SVZ astrocytic ribbon
 
Cancer Stem Cells The idea is not new
Hamburger and Salmon 1977, Science The plating efficiency of most 1 0  tumours is <1% PE =  macroscopic colonies formed number of tumour cells plated
The tumour battleground
Markers Cancer stem cells
 
 
Flow Cytometry
Fluorescence Activation Process  (or Immunofluorescence) FITC FITC Antibodies recognize specific molecules in the surface of some cells But not others When the cells are analyzed by flow cytometry the cells expressing the marker for which the antibody is specific will manifest fluorescence. Cells who lack the marker will not manifest fluorescence Antibodies are artificially conjugated to fluorochromes Antibodies FITC FITC FITC FITC
Fluorescence intensity 10 1 10 4 10 3 10 2 Relative fluorescence intensity Number of Events FITC FITC FITC FITC FITC FITC FITC FITC FITC FITC
Y X Z Y Z X Cells are presented to the laser using principles of hydrodynamic focusing  Laser optics Laser Beam Flow chamber Sheath Sample
2 Parameter Histogram FITC FL PE FL Negative Population Single Positive FITC Population Single Positive PI Population Double Positive Population
 
Xenografting
 
 
 
 
The pre-eminence of ALDH as  a cancer stem cell marker  Responsible for the oxidation (detoxification) of  cytotoxic aldehydes to carboxylic acids RCHO + NAD +  + H 2 0  RCOOH + NADH + H + : a major cellular defence mechanism
 
Vitamin A Differentiation
 
 
Charafe-Jauffret et al.
Only 21 of 203 cases were ALDH1 +
 
 
Victoria L Bautch
How do you increase CSC numbers
 
p53 MDM2 =
The cancer stem cells bandwagon
Malignant melanoma
 
 
 
In Ptc +/-  mice, Math1 + /CD15 +  (SSEA1) selects for TPCs
E-Cadherin
 
 
 
Options for therapy •  Irradiation •  Self-renewal pathways (Wnts, Notch, Hedgehog, PTEN-Akt) •  Aldehyde dehydrogenase activity •  EMT •  ABC transporters •  miRNAS and HDACi •  Targeting the niche
Cancer stem cells are very  chemoresistant and radioresistant
Front line treatment of pancreatic cancer enriches for CSCs In vitro In vivo
Colorectal xenograft treated with cyclophosphamide but  Dylla et al.
 
 
Time Selection pressures: nutrition/oxygen/immunity/therapy Clonal co-existence Clone 1 Clone 3 Clone 2
 
A more complex subclonal architecture in ALL Does genetic diversity affect the nature of tumour propagating cells? Yes, the potency, but  not  the phenotype (CD34 + CD38 -/low CD19 + )
 
Enrich Less apoptosis Enhanced DDR
Targeting L1CAM with shRNA sensitises GSCs to irradiation
 
Induce differentiation = reduced sphere formation Inhibit ALDH = increased  sphere formation
Dissociated CK20 colonosphere  cells treated  in vitro  for 6 days
 
 
 
 
 
Hedgehog signalling
Keeping things stem-like Mdm2
 
Hedgehog inhibitor: Cyclopamine Corn Lilly Cyclopia Poor oral bioavailability; short half-life
 
 
 
Reduces Gli expression Reduces ALDH bri  % * Blocked metastasis
 
 
 
GFs  PI3K  PIP3  Akt PTEN  ABCG2
Knock-down of PTEN increases efficiency of mammosphere formation
Combination therapy works
 
 
 
 
Sulforaphane
 
Related to a large reduction in ALDEFLUOR+ cells
also inhibits xenograft growth
•  Cancer is a stem cell disease •  Tumours have stem cells: ‘cancer stem cells’ •  Therapy should target cancer stem cells
 
 

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MON 2011 - Slide 16 - M.R. Alison - Joint medics and nurses spotlight session Recent developments in the field of cancer stem cells and implications in the clinical setting

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