Characterization of a carbocyanine derivative as a fluorescent penetration probe

Olive, Peggy L.; Durand, RE

doi:10.1002/cyto.990080607

Cited by 20 publications

(6 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A pioneering study using two consecutively injected perfusion markers, Hoechst 33342 (175,186) and DiOC7(3) (187,188), provided direct evidence of transient changes in blood perfusion in vivo (38), suggesting the presence of temporal fluctuations in tumor oxygenation. It inspired development of double hypoxia marker assays with consecutive injections of two different 2-nitroimidazoles for analysis of changes in hypoxia in individual experimental tumors (153,154,164,(189)(190)(191)(192).…”

Section: Dynamics Of Hypoxia: Consecutive Injection Of Hypoxia Markersmentioning

confidence: 99%

Dynamics of Tumor Hypoxia Measured with Bioreductive Hypoxic Cell Markers

et al. 2007

View full text Add to dashboard Cite

Hypoxic cells are common in tumors and contribute to malignant progression, distant metastasis and resistance to radiotherapy. It is well known that tumors are heterogeneous with respect to the levels and duration of hypoxia. Several strategies, including high-oxygen-content gas breathing, radiosensitizers and hypoxic cytotoxins, have been developed to overcome hypoxia-mediated radioresistance. However, with these strategies, an increased tumor control rate is often accompanied by more severe side effects. Consequently, development of assays for prediction of tumor response and early monitoring of treatment responses could reduce both overand undertreatment, thereby avoiding unnecessary side effects. The purpose of this review is to discuss different assays for measurement of hypoxia that can be used to detect changes in oxygen tension. The main focus is on exogenous bioreductive hypoxia markers (2-nitroimidazoles) such as pimonidazole, CCI-103F, EF5 and F-misonidazole. These are specifically reduced and bind to macromolecules in viable hypoxic cells. A number of these bioreductive drugs are approved for clinical use and can be detected with methods ranging from noninvasive PET imaging (low resolution) to microscopic imaging of tumor sections (high resolution). If the latter are stained for multiple markers, hypoxia can be analyzed in relation to different microenvironmental parameters such as vasculature, proliferation and endogenous hypoxia-related markers, for instance HIF1␣ and CA-IX. In addition, temporal and spatial changes in hypoxia can be analyzed by consecutive injection of two different hypoxia markers. Therefore, bioreductive exogenous hypoxia markers are promising as tools for development of predictive assays or as tools for early treatment monitoring and validation of potential endogenous hypoxia markers. ᭧

show abstract

Section: Dynamics Of Hypoxia: Consecutive Injection Of Hypoxia Markersmentioning

confidence: 99%

Dynamics of Tumor Hypoxia Measured with Bioreductive Hypoxic Cell Markers

et al. 2007

View full text Add to dashboard Cite

show abstract

“…MTS were then washed with cold FACS buffer and dissociated into a single cell suspension using a solution of 0.53 mM EDTA, 5% DMSO in PBS. DiOC 7 (3) is a carbocyanine derivative which penetrates poorly into MTS (Olive and Durand, 1987); thus, cells originating from the outer rim of MTS should pick up more dye and exhibit a stronger signal in the green detection channel (FL1) than cells from the center. Cells were centrifuged for 5 min at 100g, rinsed, and resuspended in cold FACS buffer containing 4 × 10 −8 M OKT9, which is above the saturating concentration for this antibody (Yazdi et al, 1995).…”

Section: Determination Of Okt9 Binding Site Distribution In Mtsmentioning

confidence: 99%

Coupled cellular trafficking and diffusional limitations in delivery of immunotoxins to multicell tumor spheroids

Wenning

Murphy

1999

Biotechnol. Bioeng.

View full text Add to dashboard Cite

Immunotoxins have the potential to be powerful tools for selective cell killing, but their lack of clinical success against solid tumors indicates a need to better understand factors which limit immunotoxin transport in three‐dimensional systems. In this work, a previously developed model which related immunotoxin toxicity to cellular trafficking in a single cell was coupled with a term accounting for diffusive transport of immunotoxin in a solid tumor sphere. This created a mathematical model which is capable of simulating the biological response of multicell tumor spheroids (MTS) to immunotoxin treatment. The model was used to predict the kinetics of protein synthesis inhibition in MTS treated with transferrin receptor‐targeted immunotoxins as a function of immunotoxin concentration and toxin choice. HeLa cells were grown as MTS and treated with immunotoxins constructed from the anti‐transferrin receptor antibody OKT9 and the toxins gelonin or CRM107, and the average protein synthesis inhibition and growth rates were measured. With no fitted parameters, the mathematical model quantitatively predicted the experimental observations. Immunotoxins were generally less effective against MTS than monolayer cells at equivalent conditions; for OKT9–gelonin at high concentrations this decrease in efficacy was attributed primarily to heterogeneous receptor distribution in MTS whereas for OKT9–CRM107 the decrease was caused primarily by a large barrier to penetration of the immunotoxin into the spheroid. The experimentally verified model was used to define the conditions which lead to large penetration barriers. In general, transport barriers in MTS become more important as immunotoxins become more effective against cells grown as monolayers. The proposed model is unique in its ability to predict toxicity in MTS directly, and is an important step toward understanding immunotoxin effect on tumors in vivo. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 62: 562–575, 1999.

show abstract

“…The bisbenzamide Hoechst 33342 (H33342, Molecular Probes, Eugene, OR, USA) was injected into the tail vein 1 min before tumour freezing, at a concentration of 7.5 mg kg Ð1 in saline (0.05 ml). This DNA-binding stain emits blue fluorescence and preferentially stains tumour cells adjacent to blood vessels (Olive and Durand, 1987;Trotter et al, 1989a). H33342 is removed very rapidly from the circulation (halflife of 2 minutes) and has been shown to be very stable once bound to DNA (Smith et al, 1988).…”

Section: Injection Of Fluorescent Stains and Ef5 Hypoxic Markermentioning

confidence: 99%

Quantification of tumour vasculature and hypoxia by immunohistochemical staining and HbO2 saturation measurements

et al. 1999

View full text Add to dashboard Cite

Tumour oxygenation has been shown to play an important role in radiotherapeutic response both in experimental tumours and in the clinic. To eliminate microregions of hypoxia, i.e. low oxygenation, numerous strategies have been used, generally aiming either to increase oxygen delivery to the tumour (Horsman et al, 1994) or to directly target the hypoxic cells using radiosensitizers or hypoxic cell cytotoxic agents (Brown and Giaccia, 1994). Several recent reports have also suggested the use of angiogenesis inhibitors (Klauber et al, 1997;OÕReilly et al, 1997) or thrombocytic agents (Huang et al, 1997) to directly target the tumour vasculature. A major shortcoming in evaluating the use of any of these approaches, however, is the inability to adequately quantify and understand the accompanying changes in tumour physiology.The current study presents a method for combining several sophisticated techniques to obtain a comprehensive two-dimensional mapping of the relationships among tumour vascular configuration, oxygen transport and hypoxic development. First, tumour oxygen availability is spatially defined by measuring intravascular blood oxygen saturations (HbO 2 ) cryospectrophotometrically in a frozen tumour block. Second, hypoxic development, in relation to the intravascular oxygen availability, is quantified in an adjacent histological section, using immunohistochemical detection of a recently developed nitroheterocyclic hypoxia marker (EF5). Third, a combination of fluorescent and immunohistological stains is used to define the distribution of distances from the viable tumour cells to the nearest anatomical or perfused blood vessel. MATERIALS AND METHODS Mice and tumour modelsThe KHT tumour, a sarcoma maintained in vivo, was passaged approximately every 2 weeks by i.m. inoculation of single-cell suspensions prepared by a mechanical dissociation procedure (Thomson and Rauth, 1974). Using 6-to 8-week-old female C3H/HeJ mice (Jackson Laboratories, Bar Harbor, ME, USA), 2×10 5 KHT cells were inoculated i.m. into the hind limbs. Tumours were selected for analysis when they reached volumes of between 320 and 1100 mm 3 (as measured by callipers (volume = πqdiam-eter 3 /6). Guidelines for the humane treatment of animals were followed as approved by the University Committee on Animal Resources. Injection of fluorescent stains and EF5 hypoxic markerLocalized areas of tumour hypoxia were assessed in frozen tissue sections by immunohistochemical identification of sites of 2-nitroimidazole metabolism. A pentafluorinated derivative (EF5) of etanidazole (synthesized by Dr R Vishnuvajjala of the National Cancer Institute) was injected i.v., 1 h before tumour freezing (0.2 ml of 10 mM EF5). Protein conjugates of EF5 have been used previously to immunize mice from which monoclonal antibodies were developed (Lord et al, 1993). These antibodies are extremely specific for the EF5 drug adducts that form when the drug is incorporated by Summary Despite the possibility that tumour hypoxia may limit radiotherapeutic response, the under...

show abstract

Characterization of a carbocyanine derivative as a fluorescent penetration probe

Cited by 20 publications

References 10 publications

Dynamics of Tumor Hypoxia Measured with Bioreductive Hypoxic Cell Markers

Dynamics of Tumor Hypoxia Measured with Bioreductive Hypoxic Cell Markers

Coupled cellular trafficking and diffusional limitations in delivery of immunotoxins to multicell tumor spheroids

Quantification of tumour vasculature and hypoxia by immunohistochemical staining and HbO2 saturation measurements

Contact Info

Product

Resources

About