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Blood vessel hyperpermeability and pathophysiology in human tumour xenograft models of breast cancer: a comparison of ectopic and orthotopic tumours.

Ho KS, Poon PC, Owen SC, Shoichet MS - BMC Cancer (2012)

Bottom Line: For these results to be meaningful, the hyperpermeable vasculature and reduced lymphatic drainage associated with tumour pathophysiology must be replicated in the model.Dextran accumulation and immunostaining results suggest that small MFP tumours best replicate the vascular permeability required to observe the EPR effect in vivo.A more predictable growth profile and the absence of ulcerated skin lesions further point to the MFP model as a strong choice for long term treatment studies that initiate after a target tumour size has been reached.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Chemical Engineering & Applied Chemistry, 200 College Street, Toronto, ON M5S 3E5, Canada.

ABSTRACT

Background: Human tumour xenografts in immune compromised mice are widely used as cancer models because they are easy to reproduce and simple to use in a variety of pre-clinical assessments. Developments in nanomedicine have led to the use of tumour xenografts in testing nanoscale delivery devices, such as nanoparticles and polymer-drug conjugates, for targeting and efficacy via the enhanced permeability and retention (EPR) effect. For these results to be meaningful, the hyperpermeable vasculature and reduced lymphatic drainage associated with tumour pathophysiology must be replicated in the model. In pre-clinical breast cancer xenograft models, cells are commonly introduced via injection either orthotopically (mammary fat pad, MFP) or ectopically (subcutaneous, SC), and the organ environment experienced by the tumour cells has been shown to influence their behaviour.

Methods: To evaluate xenograft models of breast cancer in the context of EPR, both orthotopic MFP and ectopic SC injections of MDA-MB-231-H2N cells were given to NOD scid gamma (NSG) mice. Animals with matched tumours in two size categories were tested by injection of a high molecular weight dextran as a model nanocarrier. Tumours were collected and sectioned to assess dextran accumulation compared to liver tissue as a positive control. To understand the cellular basis of these observations, tumour sections were also immunostained for endothelial cells, basement membranes, pericytes, and lymphatic vessels.

Results: SC tumours required longer development times to become size matched to MFP tumours, and also presented wide size variability and ulcerated skin lesions 6 weeks after cell injection. The 3 week MFP tumour model demonstrated greater dextran accumulation than the size matched 5 week SC tumour model (for P < 0.10). Immunostaining revealed greater vascular density and thinner basement membranes in the MFP tumour model 3 weeks after cell injection. Both the MFP and SC tumours showed evidence of insufficient lymphatic drainage, as many fluid-filled and collagen IV-lined spaces were observed, which likely contain excess interstitial fluid.

Conclusions: Dextran accumulation and immunostaining results suggest that small MFP tumours best replicate the vascular permeability required to observe the EPR effect in vivo. A more predictable growth profile and the absence of ulcerated skin lesions further point to the MFP model as a strong choice for long term treatment studies that initiate after a target tumour size has been reached.

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CD31 and αSMA co-staining. Representative images of pericytes (αSMA, violet) that are not associated with blood vessels (CD31, brown) in: A 3 week MFP, B 4 week MFP, and C 5 week SC tumours. Several blood vessels are highlighted with black arrows; blue staining represents cell nuclei. D shows that pericytes are exclusively associated with blood vessels in healthy liver control tissue. Scale bars represent 200 μm.
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Figure 4: CD31 and αSMA co-staining. Representative images of pericytes (αSMA, violet) that are not associated with blood vessels (CD31, brown) in: A 3 week MFP, B 4 week MFP, and C 5 week SC tumours. Several blood vessels are highlighted with black arrows; blue staining represents cell nuclei. D shows that pericytes are exclusively associated with blood vessels in healthy liver control tissue. Scale bars represent 200 μm.

Mentions: CD31 was also co-stained with αSMA to visualize differences in pericyte association with blood vessels. Pericytes are important blood vessel support cells that help to regulate blood flow and vessel permeability, but are often detached in tumour pathophysiology. The observed staining patterns suggest that this was the case across all tumour models (Figure 4A-C). Pericytes (violet) were distributed throughout tumour tissue instead of associating exclusively with blood vessels (brown) and forming uniform layers around the endothelial cell layer, as observed in healthy liver tissue (Figure 4D).


Blood vessel hyperpermeability and pathophysiology in human tumour xenograft models of breast cancer: a comparison of ectopic and orthotopic tumours.

Ho KS, Poon PC, Owen SC, Shoichet MS - BMC Cancer (2012)

CD31 and αSMA co-staining. Representative images of pericytes (αSMA, violet) that are not associated with blood vessels (CD31, brown) in: A 3 week MFP, B 4 week MFP, and C 5 week SC tumours. Several blood vessels are highlighted with black arrows; blue staining represents cell nuclei. D shows that pericytes are exclusively associated with blood vessels in healthy liver control tissue. Scale bars represent 200 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3539979&req=5

Figure 4: CD31 and αSMA co-staining. Representative images of pericytes (αSMA, violet) that are not associated with blood vessels (CD31, brown) in: A 3 week MFP, B 4 week MFP, and C 5 week SC tumours. Several blood vessels are highlighted with black arrows; blue staining represents cell nuclei. D shows that pericytes are exclusively associated with blood vessels in healthy liver control tissue. Scale bars represent 200 μm.
Mentions: CD31 was also co-stained with αSMA to visualize differences in pericyte association with blood vessels. Pericytes are important blood vessel support cells that help to regulate blood flow and vessel permeability, but are often detached in tumour pathophysiology. The observed staining patterns suggest that this was the case across all tumour models (Figure 4A-C). Pericytes (violet) were distributed throughout tumour tissue instead of associating exclusively with blood vessels (brown) and forming uniform layers around the endothelial cell layer, as observed in healthy liver tissue (Figure 4D).

Bottom Line: For these results to be meaningful, the hyperpermeable vasculature and reduced lymphatic drainage associated with tumour pathophysiology must be replicated in the model.Dextran accumulation and immunostaining results suggest that small MFP tumours best replicate the vascular permeability required to observe the EPR effect in vivo.A more predictable growth profile and the absence of ulcerated skin lesions further point to the MFP model as a strong choice for long term treatment studies that initiate after a target tumour size has been reached.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Chemical Engineering & Applied Chemistry, 200 College Street, Toronto, ON M5S 3E5, Canada.

ABSTRACT

Background: Human tumour xenografts in immune compromised mice are widely used as cancer models because they are easy to reproduce and simple to use in a variety of pre-clinical assessments. Developments in nanomedicine have led to the use of tumour xenografts in testing nanoscale delivery devices, such as nanoparticles and polymer-drug conjugates, for targeting and efficacy via the enhanced permeability and retention (EPR) effect. For these results to be meaningful, the hyperpermeable vasculature and reduced lymphatic drainage associated with tumour pathophysiology must be replicated in the model. In pre-clinical breast cancer xenograft models, cells are commonly introduced via injection either orthotopically (mammary fat pad, MFP) or ectopically (subcutaneous, SC), and the organ environment experienced by the tumour cells has been shown to influence their behaviour.

Methods: To evaluate xenograft models of breast cancer in the context of EPR, both orthotopic MFP and ectopic SC injections of MDA-MB-231-H2N cells were given to NOD scid gamma (NSG) mice. Animals with matched tumours in two size categories were tested by injection of a high molecular weight dextran as a model nanocarrier. Tumours were collected and sectioned to assess dextran accumulation compared to liver tissue as a positive control. To understand the cellular basis of these observations, tumour sections were also immunostained for endothelial cells, basement membranes, pericytes, and lymphatic vessels.

Results: SC tumours required longer development times to become size matched to MFP tumours, and also presented wide size variability and ulcerated skin lesions 6 weeks after cell injection. The 3 week MFP tumour model demonstrated greater dextran accumulation than the size matched 5 week SC tumour model (for P < 0.10). Immunostaining revealed greater vascular density and thinner basement membranes in the MFP tumour model 3 weeks after cell injection. Both the MFP and SC tumours showed evidence of insufficient lymphatic drainage, as many fluid-filled and collagen IV-lined spaces were observed, which likely contain excess interstitial fluid.

Conclusions: Dextran accumulation and immunostaining results suggest that small MFP tumours best replicate the vascular permeability required to observe the EPR effect in vivo. A more predictable growth profile and the absence of ulcerated skin lesions further point to the MFP model as a strong choice for long term treatment studies that initiate after a target tumour size has been reached.

Show MeSH
Related in: MedlinePlus