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Published on: February 21, 2019
Direct Observation of Endocytosis Dynamics of Anti-ErbB Modified Single Nanocargoes
1Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Tsinghua University, Beijing 100084, China.
Abstract:
The ErbB receptor family, including the epidermal growth factor receptor (EGFR) and ErbB2/3/4, regulate cell proliferation, differentiation, apoptosis, motility, etc., and their abnormalities can cause cancer and other diseases. Ligand-induced endocytosis of ErbB receptors is the key to various cancer treatment strategies, and different techniques have been developed to study this important process. Among them, single particle tracking (SPT) can reveal the spatiotemporal heterogeneity of ErbB receptors on the live cell membrane and has been used to characterize the EGFR dimerization process. Herein, we studied the endocytosis dynamics of two different ErbB receptors using dark-field microscopy. With anti-ErbB modified plasmonic gold nanorods (AuNRs) as probes, we compared the trajectories of individual anti-EGFR AuNRs (cAuNRs) and anti-ErbB AuNRs (tAuNRs) interacting with MCF-7 cells in situ in real time. The results revealed that the internalization rate of cAuNRs was faster than that of tAuNRs. Detailed SPT analysis suggests that cAuNRs enter cells through EGFR endocytosis pathway, and multiple intracellular transport modes sort the cAuNRs away from the transmembrane site. In contrast, the endocytosis resistance of ErbB2 slows down the cellular uptake rate of tAuNRs and causes some tAuNRs-ErbB2 complexes to be confined on the membrane with "circular" and "rolling circle" motions for a much longer time. Our results provide insights into the endocytosis process of the ErbB receptor family at the nanometer scale and could be potentially useful to develop cancer treatment strategies.
Insights
Single particle tracking revealed distinct endocytosis dynamics for epidermal growth factor receptor (EGFR) and ErbB2. EGFR internalization was faster, while ErbB2 showed slower uptake and membrane confinement, offering insights for cancer therapy.
Area of Science:
- Cell biology
- Nanotechnology
- Cancer research
Background:
- The ErbB receptor family (EGFR, ErbB2-4) regulates crucial cell functions, and their dysregulation is linked to cancer.
- Ligand-induced endocytosis of ErbB receptors is a critical target for cancer therapies.
- Single particle tracking (SPT) offers high-resolution insights into receptor dynamics on live cell membranes.
Purpose of the Study:
- To investigate and compare the in situ endocytosis dynamics of two ErbB receptors, EGFR and ErbB2, using gold nanorod probes.
- To elucidate the distinct intracellular trafficking pathways and membrane behaviors of EGFR and ErbB2 during endocytosis.
Main Methods:
- Utilized dark-field microscopy and single particle tracking (SPT) with anti-EGFR and anti-ErbB modified gold nanorods (AuNRs) as probes.
- Compared the real-time endocytosis trajectories of individual AuNRs interacting with MCF-7 cells.
- Analyzed nanoparticle movement patterns to infer receptor internalization rates and intracellular transport modes.
Main Results:
- Anti-EGFR AuNRs (cAuNRs) exhibited a significantly faster internalization rate compared to anti-ErbB AuNRs (tAuNRs).
- SPT analysis indicated that cAuNRs followed the EGFR endocytosis pathway with diverse intracellular sorting.
- tAuNRs targeting ErbB2 displayed slower cellular uptake and prolonged membrane confinement with specific motion patterns, suggesting endocytosis resistance.
Conclusions:
- The study reveals distinct nanometer-scale endocytosis dynamics between EGFR and ErbB2, highlighting differences in their internalization rates and intracellular trafficking.
- ErbB2's endocytosis resistance and membrane confinement behavior differ from EGFR's pathway.
- These findings provide valuable insights into ErbB receptor family dynamics and may inform the development of novel cancer treatment strategies targeting receptor endocytosis.
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