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Nanobiosensor Reports on CDK1 Kinase Activity in Tumor Xenografts in Mice
Carmen Mihaela Tîlmaciu1, Bhimareddy Dinesh2, Morgan Pellerano1
1Institut des Biomolécules Max Mousseron-CNRS, UMR5247, Université de Montpellier, Montpellier, 34093, France.
Abstract:
Probing the dynamics and quantifying the activities of intracellular protein kinases that coordinate cell growth and division and constitute biomarkers and pharmacological targets in hyperproliferative and pathological disorders remain a challenging task. Here engineering and characterization of a nanobiosensor of the mitotic kinase CDK1, through multifunctionalization of carbon nanotubes with a CDK1-specific fluorescent peptide reporter, are described. This original reporter of CDK1 activity combines the sensitivity of a fluorescent biosensor with the unique physico-chemical and biological properties of nanotubes for multifunctionalization and efficient intracellular penetration. The functional versatility of this nanobiosensor enables implementation to quantify CDK1 activity in a sensitive and dose-dependent fashion in complex biological environments in vitro, to monitor endogenous kinase in living cells and directly within tumor xenografts in mice by fluorescence imaging, thanks to a ratiometric quantification strategy accounting for response relative to concentration in space and in time.
Insights
Researchers developed a novel nanobiosensor for the mitotic kinase CDK1, enabling sensitive detection and monitoring of its activity in cells and tumors. This tool aids in understanding cell division and disease progression.
Area of Science:
- Biotechnology
- Molecular Biology
- Nanomedicine
Background:
- Intracellular protein kinases, like CDK1, regulate cell growth and division.
- Dysregulated kinase activity is implicated in hyperproliferative disorders and cancer.
- Quantifying kinase activity in vivo remains a significant challenge.
Purpose of the Study:
- To engineer and characterize a nanobiosensor for detecting the activity of the mitotic kinase CDK1.
- To develop a tool for sensitive and dose-dependent quantification of CDK1 activity in complex biological systems.
- To enable real-time monitoring of endogenous CDK1 activity in living cells and tumor xenografts.
Main Methods:
- Multifunctionalization of carbon nanotubes with a CDK1-specific fluorescent peptide reporter.
- Development of a ratiometric quantification strategy for accurate measurements.
- Application of the nanobiosensor for in vitro assays, live-cell imaging, and in vivo fluorescence imaging in mice.
Main Results:
- Successful engineering and characterization of a novel nanobiosensor for CDK1 activity.
- Demonstrated sensitive and dose-dependent quantification of CDK1 in vitro.
- Enabled monitoring of endogenous CDK1 activity in living cells and tumor xenografts via fluorescence imaging.
Conclusions:
- The developed nanobiosensor offers a sensitive and versatile platform for probing CDK1 dynamics.
- This technology facilitates the study of cell cycle regulation and kinase-targeted therapies.
- The nanobiosensor holds potential for biomarker discovery and pharmacological target validation in cancer.
