Analysis Of The Efficacy Of Radiotherapy And Related Factors For Immune Checkpoint Inhibitor-resistant Non-small Cell Lung Cancer

Part one.Analysis of the Efficacy of Radiotherapy and Related Factors for Immune Checkpoint Inhibitor-Resistant Non-Small Cell ung Cancer PurposeThis study evaluated the efficacy and related prognostic factors of radiotherapy after resistant to immune checkpoint inhibitors(ICIs)in non-small-cell lung cancer(NSCLC).To analyze the possibility of radiotherapy as a treatment option for NSCLC patients after resistant to ICIs.Materials and MethodsIn this study,data were collected from patients with NSCLC who received chest radiotherapy after resistant to ICIs at the Department of Radiation Oncology,The First Affiliated Hospital of Guangzhou Medical University from January 1,2019 to December 31,2022.The last follow-up period was up to February 28,2023.The primary endpoint was progression-free survival in patients with NSCLC treated with ICIS-resistant chest radiotherapy.SPSS 24.0 version was used for data analysis.In the second part of this study,mRNA sequencing analysis was conducted on different tumor tissues and parental tumor tissues sensitive to ICIs.Results1.In this study,a retrospective analysis was conducted on 38 patients with NSCLC who received chest radiotherapy after resistant to ICIs.Among them,31 cases(81.6%)are male,7 cases(18.4%)are female,12 cases(31.6%)are adenocarcinoma,22cases(57.9%)are squamous cell carcinoma,4 cases(10.5%)are stage Ⅱ,14 cases(36.8%)are stage Ⅲ and 20 cases(52.6%)are stage Ⅳ.2.In terms of optimal efficacy evaluation,0 patients(0%)achieved complete response,25 patients(21.2%)achieved partial response,13 patients(34.2%)achieved stable disease,and 8 patients(21.1%)achieved disease progression.The overall objective response rate was 44.7%,and the disease control rate was 78.9%.3.The local PFS of patients who continued to use ICIs after radiotherapy was 4.1months(95%CI: 2.3-5.8),which is longer than that of patients who did not continue to use ICIs after radiotherapy(95%CI: 0.0-11.117).The log-Rank test showed no significant difference in survival between the two groups(log rank P> 0.05).4.Univariate Cox regression analysis suggested that age,tumor stage,number of ICIs used before radiotherapy,ICIs used before radiotherapy,gene mutation,type of ICIs used before radiotherapy and other related factors had no correlation with the prognosis of local PFS after resistant to ICIs in patients with NSCLC(P>0.05).Tumor type and chemotherapy regimen used before radiotherapy were associated with the prognosis of local PFS after radiotherapy(P< 0.05).In terms of tumor type,the prognosis of local PFS in the adenocarcinoma group(HR=0.321,95%CI: 0.120-0.862,P< 0.05)was worse than that in the squamous cell carcinoma group.In terms of chemotherapy regimen used before radiotherapy,pemetrexed combined with platinum group(HR=0.119,95%CI: 0.024-0.596,P< 0.05),albuminbinding paclitaxel combined with platinum group(HR=0.116,95%CI: 0.021-0.636,P< 0.05)and other groups(HR=0.072,95%CI: 0.015-0.354,P< 0.05)had better prognosis of local PFS than paclitaxel combined with platinum group.In multivariate Cox regression analysis,chemotherapy regimens used before radiotherapy were associated with the prognosis of radiotherapy local PFS,albumin-binding paclitaxel combined with platinum(HR=0.063,95%CI: 0.010-0.391,P< 0.05)and other groups(HR=0.094,95%CI: 0.017-0.524,P< 0.05)compared with paclitaxel combined with platinum group,the prognosis of local PFS was better.5.After radiotherapy,12 patients(31.6%)didn’t suffer from radiation pneumonia.9 cases(23.7%)had grade I radiation pneumonia,11 cases(28.9%)had grade II radiation pneumonia,5 cases(13.2%)had grade III radiation pneumonia,1case(2.6%)had grade IV radiation pneumonia,and 0 cases(0%)had grade V radiation pneumonia.Conclusions1.Patients with NSCLC who received chest radiotherapy after resistant to ICIs can achieve better short-term efficacy.2.Use of ICIs in patients with immune checkpoint inhibitors-resistant NSCLC after chest radiotherapy does not prolong Lo PFS.Part Two.Analysis of mRNA difference in non-small cell lung cancer issues of mice which have different sensitivities to ICIs PurposeIn this study,the genes and signaling pathways related to resistant to ICIs in nonsmall cell lung cancer were screened and analyzed in mouse experimental animal models.Materials And MethodsIn this study,mRNA sequencing and comparative analysis were conducted on mouse lung non-small cell carcinoma tissues with different sensitivity to ICIs.GO enrichment analysis and KEGG enrichment analysis were performed on the screened differentially expressed mRNA to search for related genes that obvious differentially expressed.Results1.A total of 1049 differentially expressed genes are detected by mRNA sequencing in tumor tissues insensitive to ICIs compared with those sensitive to ICIs,among which 166 differentially expressed genes are up-regulated and 883 differentially expressed genes are up-regulated.The top 10 genes which have increased expression are:Pagr1a,Gcat,Krtap93,Sap25,Gm45713,Cntfr,2210017I01 Rik,Rnase2a,Cck and Gm49450.The top 10 genes which have decreased expression are:Lrp1b,Gm20431,Rsc1a1,Ccdc73,Gm49359,Gm20708 and Gm13304.In GO enrichment analysis,differential genes were mainly concentrated in biological processes such as tissue development,epithelial development,response to cytokines,and response to external stimulus.KEGG enrichment analysis suggested that differential genes were concentrated in cytokine-cytokine receptor interaction signaling pathway,viral protein interaction with cytokine and cytokine receptor signaling pathway,Interleukin-17(IL-7)signaling pathway and other signaling pathways.2.In KEGG enrichment analysis,Genes that are obviously up-regulated expressed in tumor tissues from experimental mice and have an increased expression in tissue which is sensitive to ICIs are:Cxcl12,Cxcl14,Il33,Adora1,Fabp4,Itgal1,Ttn,Ptprf,Slc36a2,Col8a2,Col28a1,Oas2,Cd209 g,Frem1,Prlr,Mpz,Fos,Tgtp1,Plin1 and Ptger3.Thbs4 was the only gene that are obviously up-regulated expressed in tumor tissues from experimental mice and have a decreased expression in tissue which is sensitive to ICIs.These genes possesses vital functions in the cytokine-cytokine receptor signaling pathway,chemokine signaling pathway,NF-kappa B signaling pathway,c AMP signaling pathway,cell adhesion moleculars signaling pathway,interleukin-17 signaling pathway,TNF signaling pathway,Wnt signaling pathway,MAPK signaling pathway,Th17 signaling pathway,Th1 and Th2 signaling pathway,T Cell receptor signaling pathway,PD-L1 expression and PD-1 checkpoint pathway in cancer signaling pathway,Toll-like receptor and other signaling pathways,which may be related to be resistant to ICIs.3.The results of Kaplan-Meier Plotter database verification showed that:1)Lung cancer patients with high expression of Il33,Col28a1,Frem1 and Prlr genes have better prognosis of OS;2)Lung cancer patients with low expression of Adora1 and Mpz genes have better prognosis of OS;3)Tumor patients with high expression of Fabp4,Ttn,Oas2,Mpz,Plin1 and Thbs4 genes can obtain better prognosis of OS from anti-PD-1 therapy;4)Tumor patients with low expression of Adora1,Ptprf,Slc36a2,Col8a2 and Fos genes can obtain better prognosis of OS from anti-PD-1 therapy.ConclusionThere were differences in gene expression in NSCLC tumor tissues with different sensitivity to ICIs,and the differentially expressed genes were mainly related to cytokine-cytokine receptor signaling pathway,chemokine signaling pathway,TNF signaling pathway,Th1 and Th2 signaling pathway,PD-L1 expression and PD-1checkpoint signaling pathway and other signaling pathways.Among the genes with distinct expression differences,some were closely related to immune system,in which Cxcl12,Cxcl14,Il33,Adora1,Fabp4,Itgal1,Ttn,Ptprf and Thbs4 may be closely related to NSCLC’s resistant to ICIS,and further research may be deserved.