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An optimized triple modality reporter for quantitative in vivo tumor imaging and therapy evaluation.

Levin RA, Felsen CN, Yang J, Lin JY, Whitney MA, Nguyen QT, Tsien RY - PLoS ONE (2014)

Bottom Line: We present an optimized triple modality reporter construct combining a far-red fluorescent protein (E2-Crimson), enhanced firefly luciferase enzyme (Luc2), and truncated wild type herpes simplex virus I thymidine kinase (wttk) that allows for sensitive, long-term tracking of tumor growth in vivo by fluorescence, bioluminescence, and positron emission tomography.Two human cancer cell lines (MDA-MB-231 breast cancer and HT-1080 fibrosarcoma cancer) were successfully transduced to express this triple modality reporter.This is the first reported use of both fluorescence and bioluminescence signals from a multi-modality reporter construct to measure drug efficacy in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, UCSD School of Medicine, University of California San Diego, La Jolla, California, United States of America.

ABSTRACT
We present an optimized triple modality reporter construct combining a far-red fluorescent protein (E2-Crimson), enhanced firefly luciferase enzyme (Luc2), and truncated wild type herpes simplex virus I thymidine kinase (wttk) that allows for sensitive, long-term tracking of tumor growth in vivo by fluorescence, bioluminescence, and positron emission tomography. Two human cancer cell lines (MDA-MB-231 breast cancer and HT-1080 fibrosarcoma cancer) were successfully transduced to express this triple modality reporter. Fluorescence and bioluminescence imaging of the triple modality reporter were used to accurately quantify the therapeutic responses of MDA-MB-231 tumors to the chemotherapeutic agent monomethyl auristatin E in vivo in athymic nude mice. Positive correlation was observed between the fluorescence and bioluminescence signals, and these signals were also positively correlated with the ex vivo tumor weights. This is the first reported use of both fluorescence and bioluminescence signals from a multi-modality reporter construct to measure drug efficacy in vivo.

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Quantification of the triple reporter optical signals to monitor therapy responses in vivo.(A) Average fluorescence signal [(p/s)/(cm2/sr)] of each MDA-MB-231 triple reporter tumor treatment group over time. (B) Average bioluminescence signal (p/s) of each MDA-MB-231 triple reporter tumor treatment group over time. (C) Average size (mm3) of each MDA-MB-231 triple reporter tumor treatment group over time based on caliper measurements. MMAE or MMAF (0.5 nmol/g) was administered on days 7, 10, 13, 16, 19, and 22. Significant decreases (p<0.005) in the tumor optical signals in the MMAE-treated group compared to the untreated and MMAF-treated groups are indicated by*.
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pone-0097415-g004: Quantification of the triple reporter optical signals to monitor therapy responses in vivo.(A) Average fluorescence signal [(p/s)/(cm2/sr)] of each MDA-MB-231 triple reporter tumor treatment group over time. (B) Average bioluminescence signal (p/s) of each MDA-MB-231 triple reporter tumor treatment group over time. (C) Average size (mm3) of each MDA-MB-231 triple reporter tumor treatment group over time based on caliper measurements. MMAE or MMAF (0.5 nmol/g) was administered on days 7, 10, 13, 16, 19, and 22. Significant decreases (p<0.005) in the tumor optical signals in the MMAE-treated group compared to the untreated and MMAF-treated groups are indicated by*.

Mentions: On day 16, a significant decrease in the tumor fluorescence signal (Figure 4A) was detected in the MMAE-treated group (1.2×108±8.2×107 [(p/s)/(cm2/sr)]) compared to the untreated (3.4×108±1.1×108 [(p/s)/(cm2/sr)]) and MMAF-treated (3.7×108±1.4×108 [(p/s)/(cm2/sr)]) groups (p = 5.0×10−4). On day 19, a significant decrease in the tumor bioluminescence signal (Figure 4B) was detected in the MMAE-treated group (1.7×108±1.3×108 (p/s)) compared to the untreated (1.0×109±5.0×108 (p/s)) and MMAF-treated (8.0×108±4.7×108 (p/s)) groups (p = 2.0×10−4). At no point did the MMAF-treated group show a significant decrease in the tumor fluorescence or bioluminescence signal compared to the untreated group.


An optimized triple modality reporter for quantitative in vivo tumor imaging and therapy evaluation.

Levin RA, Felsen CN, Yang J, Lin JY, Whitney MA, Nguyen QT, Tsien RY - PLoS ONE (2014)

Quantification of the triple reporter optical signals to monitor therapy responses in vivo.(A) Average fluorescence signal [(p/s)/(cm2/sr)] of each MDA-MB-231 triple reporter tumor treatment group over time. (B) Average bioluminescence signal (p/s) of each MDA-MB-231 triple reporter tumor treatment group over time. (C) Average size (mm3) of each MDA-MB-231 triple reporter tumor treatment group over time based on caliper measurements. MMAE or MMAF (0.5 nmol/g) was administered on days 7, 10, 13, 16, 19, and 22. Significant decreases (p<0.005) in the tumor optical signals in the MMAE-treated group compared to the untreated and MMAF-treated groups are indicated by*.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4016317&req=5

pone-0097415-g004: Quantification of the triple reporter optical signals to monitor therapy responses in vivo.(A) Average fluorescence signal [(p/s)/(cm2/sr)] of each MDA-MB-231 triple reporter tumor treatment group over time. (B) Average bioluminescence signal (p/s) of each MDA-MB-231 triple reporter tumor treatment group over time. (C) Average size (mm3) of each MDA-MB-231 triple reporter tumor treatment group over time based on caliper measurements. MMAE or MMAF (0.5 nmol/g) was administered on days 7, 10, 13, 16, 19, and 22. Significant decreases (p<0.005) in the tumor optical signals in the MMAE-treated group compared to the untreated and MMAF-treated groups are indicated by*.
Mentions: On day 16, a significant decrease in the tumor fluorescence signal (Figure 4A) was detected in the MMAE-treated group (1.2×108±8.2×107 [(p/s)/(cm2/sr)]) compared to the untreated (3.4×108±1.1×108 [(p/s)/(cm2/sr)]) and MMAF-treated (3.7×108±1.4×108 [(p/s)/(cm2/sr)]) groups (p = 5.0×10−4). On day 19, a significant decrease in the tumor bioluminescence signal (Figure 4B) was detected in the MMAE-treated group (1.7×108±1.3×108 (p/s)) compared to the untreated (1.0×109±5.0×108 (p/s)) and MMAF-treated (8.0×108±4.7×108 (p/s)) groups (p = 2.0×10−4). At no point did the MMAF-treated group show a significant decrease in the tumor fluorescence or bioluminescence signal compared to the untreated group.

Bottom Line: We present an optimized triple modality reporter construct combining a far-red fluorescent protein (E2-Crimson), enhanced firefly luciferase enzyme (Luc2), and truncated wild type herpes simplex virus I thymidine kinase (wttk) that allows for sensitive, long-term tracking of tumor growth in vivo by fluorescence, bioluminescence, and positron emission tomography.Two human cancer cell lines (MDA-MB-231 breast cancer and HT-1080 fibrosarcoma cancer) were successfully transduced to express this triple modality reporter.This is the first reported use of both fluorescence and bioluminescence signals from a multi-modality reporter construct to measure drug efficacy in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, UCSD School of Medicine, University of California San Diego, La Jolla, California, United States of America.

ABSTRACT
We present an optimized triple modality reporter construct combining a far-red fluorescent protein (E2-Crimson), enhanced firefly luciferase enzyme (Luc2), and truncated wild type herpes simplex virus I thymidine kinase (wttk) that allows for sensitive, long-term tracking of tumor growth in vivo by fluorescence, bioluminescence, and positron emission tomography. Two human cancer cell lines (MDA-MB-231 breast cancer and HT-1080 fibrosarcoma cancer) were successfully transduced to express this triple modality reporter. Fluorescence and bioluminescence imaging of the triple modality reporter were used to accurately quantify the therapeutic responses of MDA-MB-231 tumors to the chemotherapeutic agent monomethyl auristatin E in vivo in athymic nude mice. Positive correlation was observed between the fluorescence and bioluminescence signals, and these signals were also positively correlated with the ex vivo tumor weights. This is the first reported use of both fluorescence and bioluminescence signals from a multi-modality reporter construct to measure drug efficacy in vivo.

Show MeSH
Related in: MedlinePlus