Detail of Fig4A_Tg
Project
Title
In vivo bioluminescence images of transplanted E0771 tumors in HVA-transgenic mice
Description
In vivo bioluminescence images of transplanted E0771 tumors in hypoxia response element-Venus-Akaluc (HVA) transgenic (HVA-Tg) mice. Luminescence from the reporter in HVA-Tg mice allows detection of HIF activity resulting from tumor hypoxic microenvironment or inflammation. The substrate of the luciferace (AkaLumine (Aka)-HCl) was i.p. injected 10 minutes before observation. Data of three HVA-Tg mice (TgMouse1, TgMouse2, TgMouse3) are posted. The images were obtained 0 days, 8 days, 19 days, 26 days after transplantation. In the images of Day 0, Day 8 and Day 19, the leftmost mouse is a non transgenic (Non-Tg) mouse, and right three animals are HVA-Tg mice. The data of Day26 are separeted into three images.
Release, Updated
2024-12-14
License
CC BY-NC
Kind
Image data
File Formats
.TIF
Data size
11.6 MB
Organism
Mus musculus
(
NCBITaxon:10090
)
Strain(s)
C57BL/6J, B6N-Tyr^{cBrd}/BrdCrCrl
Cell Line
E0771
Datatype
-
Molecular Function (MF)
Biological Process (BP)
detection of hypoxia
(
GO:0070483
)
Cellular Component (CC)
Biological Imaging Method
luminescence detection
X scale
-
Y scale
-
Z scale
-
T scale
-
Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-
Summary of Methods
See details in Miyabara H, et. al. Cancer Sci. 2023 Oct;114(10):3935-3945.
Related paper(s)
Hitomi Miyabara, Ryuichiro Hirano, Shigeaki Watanabe, John Clyde Co Soriano, Hitomi Watanabe, Takahiro Kuchimaru, Nobuo Kitada, Tetsuya Kadonosono, Shojiro A Maki, Gen Kondoh, Shinae Kizaka-Kondoh (2023) In vivo optical imaging of tumor stromal cells with hypoxia-inducible factor activity., Cancer science
Published in 2023 Jul 22 (Electronic publication in July 22, 2023, midnight )
- doi: 10.1111/cas.15907
-
pmid: 37482942
(Abstract) Tumors contain various stromal cells, such as immune cells, endothelial cells, and fibroblasts, which contribute to the development of a tumor-specific microenvironment characterized by hypoxia and inflammation, and are associated with malignant progression. In this study, we investigated the activity of intratumoral hypoxia-inducible factor (HIF), which functions as a master regulator of the cellular response to hypoxia and inflammation. We constructed the HIF activity-monitoring reporter gene hypoxia-response element-Venus-Akaluc (HVA) that expresses the green fluorescent protein Venus and modified firefly luciferase Akaluc in a HIF activity-dependent manner, and created transgenic mice harboring HVA transgene (HVA-Tg). In HVA-Tg, HIF-active cells can be visualized using AkaBLI, an ultra-sensitive in vivo bioluminescence imaging technology that produces an intense near-infrared light upon reaction of Akaluc with the D-luciferin analog AkaLumine-HCl. By orthotopic transplantation of E0771, a mouse triple negative breast cancer cell line without a reporter gene, into HVA-Tg, we succeeded in noninvasively monitoring bioluminescence signals from HIF-active stromal cells as early as 8 days after transplantation. The HIF-active stromal cells initially clustered locally and then spread throughout the tumors with growth. Immunohistochemistry and flow cytometry analyses revealed that CD11b(+) F4/80(+) macrophages were the predominant HIF-active stromal cells in E0771 tumors. These results indicate that HVA-Tg is a useful tool for spatiotemporal analysis of HIF-active tumor stromal cells, facilitating investigation of the roles of HIF-active tumor stromal cells in tumor growth and malignant progression.
Contact
Shinae Kizaka-Kondoh
, Tokyo Institute of Technology
, School of Life Science and Technology
Contributors
Hitomi Miyabara , Shigeaki Watanabe, Hitomi Watanabe, Gen Kondoh, Takahiro Kuchimaru
OMERO Project
Source