Cell And Organoid Sensors For Anti-lung Cancer Drug Evaluation And Individualized Treatment

Lung cancer currently causes the most deaths in the world.It is a common hope for people worldwide to find more effective treatments and develop new anti-lung cancer drugs.However,traditional drug development process takes long period and large cost,while the success rate is low.There are very few drug candidates eventually get the approval and reach the market.In order to improve the success rate of drug development,it is essential to find more bionic models and more efficient methods for drug screening.At the same time,the heterogeneity of lung cancer is extremely obvious.Individual characteristics of different patients are significantly different.The efficacy of universal platinum-based dual-drug chemotherapy has reached a bottleneck.With the development of molecular biology and high-throughput gene sequencing technology,individualized treatment is an inevitable trend in lung cancer treatment in the future.Therefore,it is essential to find a better approach to guide individualized treatment and improve patients’survival.Based on patient-derived models and biosensing technology,this article carried out some innovative researches on cell and organoid sensors for drug screening and individualized treatment.First,an electrochemical cell sensor for cellular reactive oxygen species detection and a multifunctional cell sensor for comprehensively evaluating drug-induced inhibition and cardiotoxicity were designed.Subsequently,lung cancer patient-derived cells and 3D impedance sensors were used to predict drug sensitivities of specific patients.Finally,patient-derived lung cancer organoids,3D myocardial structures,and liver organoids were established.By integrating organoids with organoid sensors,the multi-organoids sensor system was constructed studied for the individualized treatment of lung cancer.The main research contents and innovative work of this paper include:1.Proposed the design method of a electrochemical cell sensor for detecting intracellular reactive oxygen species,which can be used for the screening of anti-lung cancer drugs and related basic researches.Reactive oxygen species（ROS）plays an instrumental role in lung cancer development.Many anti-lung cancer drugs work by targeting intracellular ROS.In this paper,a simple and disposable electrochemical cell sensor for ROS detection was designed and manufactured.Graphene（RGO）and platinum nanoparticles（Pt NPs）were modified on the surface of the screen-printed electrode by electrodeposition method to construct an electrochemical sensor,which can detect H₂O₂ with high sensitivity and selectivity in the range of 1 to 10μM.The detection limit of the electrochemical cell sensor was 0.65μM.Then methacrylic acid anhydride gelatin/graphene hydrogel（Gel MA/RGO）was used to immobilize A549 lung cancer cells on the surface of the sensor.The Gel MA/RGO hydrogel can maintain cell survival on the electrode.The sensor can detect the increase of intracellular ROS induced by drugs（such as f MLP）or be used for screening drugs that target ROS.Results showed that honokiol（a natural polyphenol）induces the death of lung cancer cells A549 by up-regulating the intracellular ROS.Confocal fluorescence microscopy and flow cytometry verify the reliability of the sensor test results,indicating that the electrochemical cell sensor can be applied to related drug screening and basic research.2.Proposed the design method of a multifunctional biosensor,which can be used for comprehensively evaluating drug-induced lung cancer death and cardiotoxicity.In lung cancer therapy,drug-induced cardiotoxicity is a prominent problem.In this paper,a novel multifunctional biosensor was designed and manufactured.The interdigital electrodes were used to measure cell survival and beating while the microelectrodes were used for extracellular field potential.The multifunctional biosensor could measure the drug’s effect on lung cancer viability and cardiomyocyte function comprehensively.Using the novel biosensor,we evaluated the efficacy and cardiotoxicity of AC0010,a novel tyrosine kinase inhibitor targeting the EGFR T790M mutation.The EGFR T790M mutation contributes to the acquired resistance of lung cancer patients who received targeted therapy.Results demonstrated that AC0010 significantly inhibited EGFR T790M mutant lung cancer cells and impaired cardiomyocytes’function.The AC0010 affected the action potential and beating possibly by regulating the function of ion channels and sarcoplasmic reticulum Ca²⁺pump.Increased LDH release and oxygen stress were also detected after AC0010treatment.As AC0010 was still under clinical trials,this paper could guide the trials and applications in the future.3.Proposed a drug sensitivity detection method based on lung cancer patient-derived cells and 3D cellular sensors,which may guide the individualized treatment of lung cancer patients.The heterogeneity of lung cancer leads to serious individual differences.Drug sensitivity tests before treatment can improve the success rate.This paper proposed a drug sensitivity testing approach based on patient-derived cells and 3D cellular sensors.The drug sensitivities to cisplatin,honokiol,and vandetanib of 2D and 3D cultured lung adenocarcinoma patient-derived cells（PDCs）were detected by 2D and3D cellular impedance sensors.Fluorescent staining test and in vivo study were conducted for validations.Results showed that 3D cultured PDC has stronger resistance than 2D cultured cells.2D and 3D cellular impedance sensors can distinguish the sensitivities of three drugs in 2D and 3D culture environments.The drug sensitivity evaluation approach proposed in this paper was expected to guide the individualized treatment of lung cancer patients.Doctors can use 2D cellular impedance sensors for preliminary screening of a large number of drugs,and then use3D cell impedance sensors for secondary screening to select the most promising drug for individualized treatment.4.Proposed the design method of a multi-organoid sensor system by integrating multiple organoids and biosensors,which can predict patient’s response to drugs and guide the individualized treatment.Drugs go through complex metabolic processes in patients’bodies.With the power of reconstructing patients’responses,organoids help doctors make pharmacotherapeutic schemes for individualized treatment.In this paper,a multi-organoid sensor system was designed by combining multiple organoid models（lung cancer organoids,3D myocardial structures,and liver organoids）and multiple sensors（organoid impedance sensors and multifunctional biosensors）.The multiple organoid sensor systems that can reconstruct patients’responses to drugs were expected to guide the individualized treatment of lung cancer patients.The multi-organoid sensor system simulates the liver-heart-lung axis metabolic process of oral drugs in the body by constructing a flow path system that communicates liver organoid,3D myocardial structure,and lung cancer organoid.The biomimetic design can predict drug efficacy,cardiotoxicity,and hepatoxicity for patients.The organoid impedance sensor could measure drug-induced inhibition on lung cancer organoids and liver organoids,and the multifunctional biosensor can measure the cardiotoxicities.The initial results showed that lung cancer organoids have stronger drug resistance to cisplatin,docetaxel,and AC0010,which could be distinguished by the organoid impedance sensor.HCC827 organoids carrying EGFR mutations were more sensitive to the targeted drug AC0010 than wide-type lung cancer organoids,manifesting as a lower IC₅₀.The multi-organoid sensor system can stimulate the in vivo physiological environment.To a certain extent,the system might substitute for animal experiments and guide the individualized treatment of lung cancer patients.