High-throughput Drug Screen For Patient Derived Xenografts/cells Of Head And Neck Cancer Based On Date Annotation

PURPOSE:Head and neck squamous cell carcinoma(HNSCC)is the major type of head and neck cancer and it is the sixth most common cancer worldwide.Despite improved survival,especially through the introduction of targeted agents,there are still no ideal preclinical models and clear biomarkers used as guides for clinical targeted drugs or chemotherapy drugs.Therefore,the phenomenon of blind medication emerges.Cetuximab is the only drugs approved by the FDA for HNSCC,but it has a certain effect in only about 10%-15%of patients.In recent years,clinical drug resistance of cetuximab has become more and more obvious.More importantly,most of the early clinical trial tests for drugs failed.Although tumor cell lines are an important tool for evaluating preclinical drugs,none of these cell lines can effectively predict clinical outcomes.This research aims to establish patient-derived xenograft(PDX)and patient-derived cell(PDC)annotated with a comprehensive data of genetic and clinical information.We provide more alternative treatments for head and neck squamous cell carcinoma through in vitro targeted and chemotherapeutic drugscreen of a variety of PDCs and preclinical PDX validation.METHODS:1.Tumor specimens of Head and Neck Squamous Carcinoma were collected and subcutaneously implanted into the immune-compromised nude mice.PDX of HNSCC was constructed by serial passage and PDC was isolated by the patient’s primary tissue or the patient’s corresponding original three generations of PDX.The stability of the constructed PDX/PDC was evaluated by histopathology and bioinformatics comparison2.Appropriate drug candidates were picked out by screening for PDC.Through in vitro cell-based drug test,we observed the differences in sensitivity of different patient-derived cells to the same drugand its effect on changes in phenotypes.3.Using the constructed PDX models to verify the candidate drugs screened by the PDC models,and observe the effect of the inhibitor on the transplanted tumor by immunohistochemical staining and tunnel fluorescent staining.RESULTS:1.Through primary tumor implantation and sequencing passage,we successfully constructed 22 head and neck squamous cell carcinoma xenograft models and corresponding patient-derived cell lines.the stability of PDX/PDC was evaluated by histopathology and bioinformatics,we found that PDX/PDC retained the biological behavior and genetic characteristics of the matchedprimary tumor.2.We found CHK1inhibitor PY34 which was more sensitive than other agents in major patient derived tumor cellsusing high-throughput drug screening in vitro.We selected two relatively sensitive PDC models and found that the PY34 could significantly inhibit the formation of cell clonal formation,increase apoptotic cells and inhibit downstream signaling pathway at low concentrations.On the contrary,it has been found that the above-mentioned phenomenon can only be formed under the action of a high concentration of PY34 within insensitive PDCs.Similarly,we observed the same phenomenon in relatively sensitive and insensitive traditional commercial cell lines of HNSCC.3.We used the PDX model corresponding to the selected cells for in vivo verification,and found that the No.173 PDX model corresponding to PDC relatively sensitive to CHK1 inhibitor showed significant inhibition on tumor growth,downregulatd tumor proliferation and induced apotosis cells.The No.044 PDX model corresponding to PDC,which was relatively insensitive in vitro,had no obvious tumor inhibition effect under the action of CHK1 inhibitor.It is also obvious that there was no statistical difference in tumor proliferation and induced apotosis cells in the relatively insensitive samples.CONCLUSIONS:Using high-throughput drug screen for PDC of HNSCC with comprehensive stability assessment by histopathology and bioinformatics,we found a new suitable candidate targeted drug CHK1 inhibitor PY34 and verified the mechanism of action of PY34.The corresponding PDX model was used to evaluate the efficacy of the drug in vivo,providing more treatment options for HNSCC and propose new ideas for the development of other oncology drugs.