In vivo measurement of gene expression, angiogenesis and physiological function in tumors using multiphoton laser scanning microscopy

Abstract

Intravital microscopy coupled with chronic animal window models has provided stunning insight into tumor pathophysiology, including gene expression, angiogenesis, cell adhesion and migration, vascular, interstitial and lymphatic transport, metabolic microenvironment and drug delivery. However, the findings to date have been limited to the tumor surface (< 150 microm). Here, we show that the multiphoton laser-scanning microscope can provide high three-dimensional resolution of gene expression and function in deeper regions of tumors. These insights could be critical to the development of novel therapeutics that target not only the tumor surface, but also internal regions.

Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice

“…Therefore, they may constitute a useful preclinical model for testing angiogenesis inhibitors for treatment of cancers, macular degeneration, diabetic retinopathy, and chronic inflammatory diseases involving neovascularization. In combination with intravital microscopy, which can provide high resolution in three dimensions in deeper tissues of the live animal (Brown et al, 2001;Jain et al, 2002), the Flk1::H2B-EYFP mouse could provide a readout in real time in a number of preclinical models of human cancer. These transgenic mice will, therefore, create new possibilities for the study of normal and aberrant vascular development from embryo to adult and in models for human disease.…”

Dynamic imaging of cancer growth and invasion: a modified skin-fold chamber model

“…Invasion promoting factors reside in the migratory capability of cancer cells, the release of invasion-enhancing factors from the adjacent tumor stroma, and the metabolic and perfusion state of the lesion (Brown et al 2001;Condeelis and Segall 2003;Gaggioli et al 2007;Nakamura et al 2007). The tumor-stroma cross-talk and its eVect on cancer invasion are only incompletely recapitulated by in vitro models, which often lack the anatomy and cellular composition of the tumor environment ).…”

“…To directly visualize cancer progression, including cell invasion, interaction with the adjacent stroma and neovessel formation, multiphoton microscopy allows long-term imaging at acceptable penetration depth (few hundred micrometer), and limited photobleaching, and -toxicity (Brown et al 2001;Condeelis and Segall 2003;Helmchen and Denk 2005). The dorsal skinfold chamber is a widely used in vivo model for preclinical cancer research for drug testing, angiogenesis and tumor progression studies, as well as vascular leakage and intratumoral pressure measurements (Asaishi et al 1981;Boucher et al 1996;Guba et al 2002;Leunig et al 1992;ReyesAldasoro et al 2008).…”

Role of p53 and Rb in Ovarian Cancer

“…MPM has been used to help answer diverse biological questions such as how gene expression correlates with metastasis, whether senile plaques change size in a mouse model of Alzheimer's disease and the role of sensory deprivation in cortical plasticity (Brown et al, 2001;Christie et al, 2001;Lendvai et al, 2000;Wang et al, 2002). In addition to low phototoxicity, MPM can allow analysis of individual cell migration and motility in a time-lapse manner (FleskenNikitin et al, 2005), while long-term, repeated imaging procedures may be carried out by performing MPM during several rounds of survival surgery (Christie et al, 2001), allowing one to closely follow development of EOC from the very earliest stages of carcinogenesis.…”