Experimental pathology by intravital microscopy and genetically encoded fluorescent biosensors

Terai, Kenta

doi:10.1111/pin.12925

Cited by 8 publications

(6 citation statements)

References 105 publications

(110 reference statements)

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“…Additional information can be obtained during imaging by visualizing parts of the microenvironment, such as the collagen‐1 fibers through second‐harmonic generation (SHG), the vascular system using fluorescently labeled dextrans, and immune cell subsets using specific fluorescent antibodies (Headley et al., 2016; Ritsma et al., 2012). Moreover, molecular signaling dynamics in the tumor cells can be visualized using fluorescent reporters (Giampieri, Pinner, & Sahai, 2010; Matsuda & Terai, 2020; Muta et al., 2018). Through this approach, alterations in signaling pathways and cellular behavior can be studied at high resolution, in vivo, over time, providing unique insights into the dynamic metastatic process as well as responses to therapies (Margarido, Bornes, Vennin, & van Rheenen, 2019).…”

Section: Commentarymentioning

confidence: 99%

An Experimental Liver Metastasis Mouse Model Suitable for Short and Long‐Term Intravital Imaging

2021

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The liver is a frequent site of cancer metastasis, but current treatment options for cancer patients with liver metastasis are limited, resulting in poor prognosis. Colonization of the liver by cancer cells is a multistep and temporally controlled process. Investigating this process in biological relevant settings in a dynamic manner may lead to new therapeutic avenues. Experimental mouse models of liver metastasis combined with high-resolution microscopy methods can facilitate study of the mechanisms that underlie the outgrowth of cancer cells in the liver. Intravital imaging can provide information on the behavior of tumor cells in their biological setting, in time frames of hours to days. In this unit, we describe the experimental induction of liver metastasis through administration of cancer cells into mice via mesenteric vein injection. The behavior of these injected cells can then be studied using intravital imaging by surgical exposure or through an abdominal imaging window. The approach is described for use with an upright multiphoton microscope, making it widely applicable.

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Section: Commentarymentioning

confidence: 99%

An Experimental Liver Metastasis Mouse Model Suitable for Short and Long‐Term Intravital Imaging

2021

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“…Applying real-time in vivo imaging to assess tumor progression in its local environment has revealed time and spatial dependence of drug targeting vulnerabilities in various cancers (Conway et al, 2017;Ebrahim and Weigert, 2019;Ellenbroek and van Rheenen, 2014;Fruhwirth et al, 2011;Nobis et al, 2018;Scheele et al, 2016). To this end, we and others have employed the use of Fo ¨rster resonance energy transfer (FRET) biosensor mice to explore a wide range of proteins involved in cancer pro-gression and metastasis and their pharmacological inhibition (Festy et al, 2007;Hirata et al, 2015;Hiratsuka et al, 2015;Matsuda and Terai, 2020). For example, we have assessed fluctuations and targeting of the non-receptor tyrosine kinase Src during cancer invasion (Nobis et al, 2013), CDK1 activity in response to chemotherapy and extracellular matrix (ECM) manipulation (Vennin et al, 2017(Vennin et al, , 2019, and Akt activity in response to drug targeting in moving hypoxic regions of live tumors (Conway et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Optimizing metastatic-cascade-dependent Rac1 targeting in breast cancer: Guidance using optical window intravital FRET imaging

Floerchinger¹,

Murphy²,

Latham³

et al. 2023

Preprint

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<p>Assessing drug response within live native tissue provides increased fidelity with regards to optimizing efficacy while minimizing off-target effects. Here, using longitudinal intravital imaging of a Rac1-Forster resonance energy transfer (FRET) biosensor mouse coupled with in vivo photoswitching to track intratumoral movement, we help guide treatment scheduling in a live breast cancer setting to impair metastatic progression. We uncover altered Rac1 activity at the center versus invasive border of tumors and demonstrate enhanced Rac1 activity of cells in close proximity to live tumor vasculature using optical window imaging. We further reveal that Rac1 inhibition can enhance tumor cell vulnerability to fluid-flow-induced shear stress and therefore improves overall anti-metastatic response to therapy during transit to secondary sites such as the lung. Collectively, this study demonstrates the utility of single-cell intravital imaging in vivo to demonstrate that Rac1 inhibition can reduce tumor progression and metastases in an autochthonous setting to improve overall survival. </p>

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“…The invention of the microscope leads the early pioneers to observe living cells in the seventeenth century [ 8 ]. Despite this long history, it was only after the widespread application of a two-photon excitation microscope (2PEM) that the number of studies using IVM started to increase [ 9 , 10 ]. Two-photon excitation arises from the simultaneous absorption of two photons in a single event, which depends on the square of the light intensity.…”

Section: Introductionmentioning

confidence: 99%

In vivo imaging of inflammatory response in cancer research

Konishi

Terai

2023

Inflamm Regener

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Inflammation can contribute to the development and progression of cancer. The inflammatory responses in the tumor microenvironment are shaped by complex sequences of dynamic intercellular cross-talks among diverse types of cells, and recapitulation of these dynamic events in vitro has yet to be achieved. Today, intravital microscopy with two-photon excitation microscopes (2P-IVM) is the mainstay technique for observing intercellular cross-talks in situ, unraveling cellular and molecular mechanisms in the context of their spatiotemporal dynamics. In this review, we summarize the current state of 2P-IVM with fluorescent indicators of signal transduction to reveal the cross-talks between cancer cells and surrounding cells including both immune and non-immune cells. We also discuss the potential application of red-shifted indicators along with optogenetic tools to 2P-IVM. In an era of single-cell transcriptomics and data-driven research, 2P-IVM will remain a key advantage in delivering the missing spatiotemporal context in the field of cancer research.

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Experimental pathology by intravital microscopy and genetically encoded fluorescent biosensors

Cited by 8 publications

References 105 publications

An Experimental Liver Metastasis Mouse Model Suitable for Short and Long‐Term Intravital Imaging

An Experimental Liver Metastasis Mouse Model Suitable for Short and Long‐Term Intravital Imaging

Optimizing metastatic-cascade-dependent Rac1 targeting in breast cancer: Guidance using optical window intravital FRET imaging

In vivo imaging of inflammatory response in cancer research

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