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Un biosensore electroquímico para la detección in situ de calreticulina para evaluar la muerte celular inmunogénica
Zhenguo Zhang1, Dongfang Ren1, Shushen Li1
1Center for Cell Structure and Function, Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Sciences, Shandong Normal University, Jinan, Shandong 250014, China.
Insights
Los investigadores desarrollaron un biosensore electroquímico que utiliza eZIF-8@MB y rGO-MWCNT para detectar cambios en la calreticulina (CRT), lo que permite un análisis preciso de las etapas de muerte celular inmunogénica (ICD) para la evaluación de la terapia contra el cáncer.
Área de la Ciencia:
- Ingeniería Biomédica
- Nanotecnología
- Investigación sobre el Cáncer
Sus antecedentes:
- Las inmunoterapias que inducen la muerte celular inmunogénica (ICD) son prometedoras para el tratamiento del cáncer.
- La detección de la progresión de la ICD es crucial para evaluar la eficacia terapéutica.
- Los métodos actuales para analizar las etapas de la ICD son limitados.
Objetivo del estudio:
- Desarrollar un biosensore electroquímico novedoso para detectar cambios en la calreticulina (CRT) durante la ICD.
- Analizar diferentes etapas de la ICD en modelos celulares y in vivo.
- Proporcionar una herramienta para evaluar las terapias contra el cáncer basadas en la ICD.
Principales métodos:
- Fabricación de un nanocomposite de marco de imidazolato de zinc etado (eZIF-8@MB) y óxido de grafeno reducido-nanotubo de carbono multi-pared (rGO-MWCNT).
- Desarrollo de un biosensore electroquímico para detectar la CRT de la superficie celular.
- Establecimiento y confirmación de modelos celulares y de ratón de ICD.
- Caracterización de la morfología y el rendimiento del nanocomposite.
- Voltamperometría de pulso diferencial para la detección de CRT.
Principales resultados:
- El nanocomposite eZIF-8@MB/rGO-MWCNT exhibió un excelente rendimiento.
- El biosensore demostró una detección rápida, sensible y estable de CRT.
- Se observó una relación lineal para la detección de CRT en el rango de 10^2 a 10^6 células/mL.
- El biosensore detectó con éxito etapas diferenciales de CRT en un modelo de ratón de ICD.
Conclusiones:
- El biosensore electroquímico desarrollado permite un análisis preciso de la progresión de la ICD.
- Esta tecnología ofrece una herramienta novedosa para evaluar las inmunoterapias contra el cáncer basadas en la ICD.
- El estudio facilita el cribado de agentes farmacéuticos dirigidos a la ICD.
Abstract:
Immunotherapies centered on inducing immunogenic cell death (ICD) hold promise for overcoming the limitations of current conventional cancer treatments and enable the maximal eradication of residual tumor cells within patients. In this study, by using the unique performance of a porous etched zeolitic imidazolate framework-8 incorporating a methylene blue (eZIF-8@MB) probe and a reduced graphene oxide-multiwalled carbon nanotube (rGO-MWCNT) nanocomposite, we established an electrochemical biosensor that can specifically detect the changes of calreticulin (CRT) on living cells and analyze different stages of ICD. The ICD cellular model was established and confirmed via various conventional techniques. The morphology and performance of the nanocomposite were thoroughly characterized. Differential pulse voltammetry revealed the rapid, sensitive, and stable responses of CRT with a good linear relationship within the cell concentration range of 102 to 106 cells mL-1. We successfully constructed the ICD mouse model and applied the biosensor to detect differential CRT stages across cancer tissues. This study facilitates the applications of electrochemical sensing techniques in the precise analysis of the progress of ICD, thereby offering a novel method and tool for evaluating ICD-based cancer therapies and screening related pharmaceutical agents.
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