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In situ drug-receptor binding kinetics in single cells: a quantitative label-free study of anti-tumor drug resistance.

Wang W, Yin L, Gonzalez-Malerva L, Wang S, Yu X, Eaton S, Zhang S, Chen HY, LaBaer J, Tao N - Sci Rep (2014)

Bottom Line: Many drugs are effective in the early stage of treatment, but patients develop drug resistance after a certain period of treatment, causing failure of the therapy.We further showed that the steric hindrance of Mucin-4, a membrane protein, was responsible for the altered drug-receptor binding.This effect of a third molecule on drug-receptor interactions cannot be studied using traditional purified protein methods, demonstrating the importance of the present intact cell-based binding kinetics analysis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.

ABSTRACT
Many drugs are effective in the early stage of treatment, but patients develop drug resistance after a certain period of treatment, causing failure of the therapy. An important example is Herceptin, a popular monoclonal antibody drug for breast cancer by specifically targeting human epidermal growth factor receptor 2 (Her2). Here we demonstrate a quantitative binding kinetics analysis of drug-target interactions to investigate the molecular scale origin of drug resistance. Using a surface plasmon resonance imaging, we measured the in situ Herceptin-Her2 binding kinetics in single intact cancer cells for the first time, and observed significantly weakened Herceptin-Her2 interactions in Herceptin-resistant cells, compared to those in Herceptin-sensitive cells. We further showed that the steric hindrance of Mucin-4, a membrane protein, was responsible for the altered drug-receptor binding. This effect of a third molecule on drug-receptor interactions cannot be studied using traditional purified protein methods, demonstrating the importance of the present intact cell-based binding kinetics analysis.

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Related in: MedlinePlus

Herceptin interactions with SK-BR3 Herceptin-resistant (H6 and C5) and SK-BR3 Herceptin-sensitive (E8 and C11) clones.A) The average SPR sensorgram of each clone and the corresponding two-constant (H6 and C5) or one-constant (E8 and C11) fitting. B) The resistance index (RI) map of each clone, demonstrating the potency of individual cell being resistant to Herceptin treatment. C) SPR sensorgrams of two individual cells are shown to demonstrate the cell-to-cell variation when interacts with Herceptin. Scale bars, 100 μm.
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f4: Herceptin interactions with SK-BR3 Herceptin-resistant (H6 and C5) and SK-BR3 Herceptin-sensitive (E8 and C11) clones.A) The average SPR sensorgram of each clone and the corresponding two-constant (H6 and C5) or one-constant (E8 and C11) fitting. B) The resistance index (RI) map of each clone, demonstrating the potency of individual cell being resistant to Herceptin treatment. C) SPR sensorgrams of two individual cells are shown to demonstrate the cell-to-cell variation when interacts with Herceptin. Scale bars, 100 μm.

Mentions: The unique capability for determining drug-receptor binding kinetics in intact cells was applied to study Herceptin resistance mechanism by comparing the Herceptin binding kinetics of Herceptin-sensitive with that of Herceptin-resistant cells. Two Herceptin-sensitive (C11 and E8) and two Herceptin-resistant (H6 and C5) SK-BR3 clones were studied (Supplementary Information Section 5) at a Herceptin concentration of 2.1 μg/mL, which is relevant to the drug concentration in the body used in clinical therapy. The sensorgrams of the four clones are shown in Fig. 4A. The Herceptin-resistant clones, H6 and C5, exhibit two distinct dissociation processes, a fast process followed by a slow process (Figs. 4A, H6 and C5). This behavior is completely different from the dissociation processes in the Herceptin-sensitive clones (Figs. 4A, E8 and C11) and in the other Her2-positive cell lines (Figs. 2D, 3A and 3B), which follow the simple single dissociation kinetics.


In situ drug-receptor binding kinetics in single cells: a quantitative label-free study of anti-tumor drug resistance.

Wang W, Yin L, Gonzalez-Malerva L, Wang S, Yu X, Eaton S, Zhang S, Chen HY, LaBaer J, Tao N - Sci Rep (2014)

Herceptin interactions with SK-BR3 Herceptin-resistant (H6 and C5) and SK-BR3 Herceptin-sensitive (E8 and C11) clones.A) The average SPR sensorgram of each clone and the corresponding two-constant (H6 and C5) or one-constant (E8 and C11) fitting. B) The resistance index (RI) map of each clone, demonstrating the potency of individual cell being resistant to Herceptin treatment. C) SPR sensorgrams of two individual cells are shown to demonstrate the cell-to-cell variation when interacts with Herceptin. Scale bars, 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4196117&req=5

f4: Herceptin interactions with SK-BR3 Herceptin-resistant (H6 and C5) and SK-BR3 Herceptin-sensitive (E8 and C11) clones.A) The average SPR sensorgram of each clone and the corresponding two-constant (H6 and C5) or one-constant (E8 and C11) fitting. B) The resistance index (RI) map of each clone, demonstrating the potency of individual cell being resistant to Herceptin treatment. C) SPR sensorgrams of two individual cells are shown to demonstrate the cell-to-cell variation when interacts with Herceptin. Scale bars, 100 μm.
Mentions: The unique capability for determining drug-receptor binding kinetics in intact cells was applied to study Herceptin resistance mechanism by comparing the Herceptin binding kinetics of Herceptin-sensitive with that of Herceptin-resistant cells. Two Herceptin-sensitive (C11 and E8) and two Herceptin-resistant (H6 and C5) SK-BR3 clones were studied (Supplementary Information Section 5) at a Herceptin concentration of 2.1 μg/mL, which is relevant to the drug concentration in the body used in clinical therapy. The sensorgrams of the four clones are shown in Fig. 4A. The Herceptin-resistant clones, H6 and C5, exhibit two distinct dissociation processes, a fast process followed by a slow process (Figs. 4A, H6 and C5). This behavior is completely different from the dissociation processes in the Herceptin-sensitive clones (Figs. 4A, E8 and C11) and in the other Her2-positive cell lines (Figs. 2D, 3A and 3B), which follow the simple single dissociation kinetics.

Bottom Line: Many drugs are effective in the early stage of treatment, but patients develop drug resistance after a certain period of treatment, causing failure of the therapy.We further showed that the steric hindrance of Mucin-4, a membrane protein, was responsible for the altered drug-receptor binding.This effect of a third molecule on drug-receptor interactions cannot be studied using traditional purified protein methods, demonstrating the importance of the present intact cell-based binding kinetics analysis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.

ABSTRACT
Many drugs are effective in the early stage of treatment, but patients develop drug resistance after a certain period of treatment, causing failure of the therapy. An important example is Herceptin, a popular monoclonal antibody drug for breast cancer by specifically targeting human epidermal growth factor receptor 2 (Her2). Here we demonstrate a quantitative binding kinetics analysis of drug-target interactions to investigate the molecular scale origin of drug resistance. Using a surface plasmon resonance imaging, we measured the in situ Herceptin-Her2 binding kinetics in single intact cancer cells for the first time, and observed significantly weakened Herceptin-Her2 interactions in Herceptin-resistant cells, compared to those in Herceptin-sensitive cells. We further showed that the steric hindrance of Mucin-4, a membrane protein, was responsible for the altered drug-receptor binding. This effect of a third molecule on drug-receptor interactions cannot be studied using traditional purified protein methods, demonstrating the importance of the present intact cell-based binding kinetics analysis.

Show MeSH
Related in: MedlinePlus