3 Zhenhe Technology and 3 Shanghai Pulmonary Hospital affiliated to Tongji University -Professor Zhou Caicun Research results of the team collaboration of Professor Zhou Caicun were published in "Cancer Communications", IF: 15.283. In this study, multiple immunofluorescence (mIF) detection was performed using and rontis products.
PD-1 inhibitor combined with chemotherapy has become a new standard for first-line treatment for patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) , but nearly half of patients still cannot benefit from this combination therapy. To further expand the benefit population of PD-1 inhibitor combined with chemotherapy regimen, it is important to find reliable predictive biomarkers. However, neither PD-L1 expression nor single biomarkers such as TMB could predict the efficacy of combined treatment. Therefore, this study explored the predictive value of the rational combination of different markers on the efficacy of PD-1 inhibitor combined with chemotherapy in patients with advanced NSCLC by detecting the expression levels of tumor immune microenvironment biomarkers.
study overview
Patient situation
Patient situation
A total of 144 patients with stage IIIB-IV non-squamous NSCLC who were initially treated, 77 of which received PD-1 inhibitor carilizumab combined with chemotherapy, and 67 were treated with chemotherapy only. The objective response rate of patients with carilizumab combined with chemotherapy group (ORR) was significantly higher than that of patients with chemotherapy group (59.7% vs 37.3%, P=0.007).
Sample detection
Using Zhenhe Landis product , based on multiple immunofluorescence (mIF) technology, the tissue samples before treatment immunomicroenvironmental biomarkers (including PDL1, CD8, CD68, CD4 and FOXP3) were detected and analyzed.
Table 1 Clinical baseline characteristics of 144 NSCLC patients
Key Study results
Co-expression levels of respondents in the chemotherapy group were significantly higher than those in the non-responders, and no significant difference was found in the chemotherapy group.
This study first evaluated the correlation between the expression levels of single and co-expression markers of (positive cell density and percentage of positive cells) and objective remission in the carrilizumab combined with chemotherapy group.
single marker (PD-L1, CD8, CD68, CD4, FOXP3)
PD-L1 positive cell density (P=0.013) and percentage of positive cells (P=0.020) , CD8 positive cell density (P=0.046) The percentage of and CD68 positive cells (P=0.048) were significantly higher than those who did not respond. The expression levels of CD4 and FOXP3 have no correlation with objective remission (Figure 1-B) .
Figure 1-B Correlation between single marker expression level and therapeutic efficacy
co-expression marker (CD8/PD-L1, CD68/PD-L1, CD4/FOXP3, CD8/CD68/PD-L1, CD4/FOXP3/PD-L1)
responders had significantly higher CD8/PD-L1 and CD68/PD-L1 positive cell density and percentage of positive cells than those of non-responders, while the co-expression levels of CD4/FOXP3, CD8/CD68/PD-L1 and CD4/FOXP3/PD-L1 were similar between responders and non-responders.
Figure 1-C Correlation between co-expression marker expression levels and therapeutic efficacy
3 carelilizumab combined with chemotherapy group, the ORR of patients with high co-expression levels of CD8/PD-L1 or CD68/PD-L1 was significantly higher; the PFS and OS of patients with high co-expression groups of CD8/PD-L1 and CD68/PD-L1 were significantly improved.
Next, this study evaluated the correlation between coexpression markers and clinical outcomes in the carrilizumab combined with chemotherapy group.The results showed that the high CD8/PD-L1 and high CD68/PD-L1 co-expression levels of had better predictive value . In the
CD8/PD-L1 co-expression group
3carelilizumab combined with chemotherapy group, the ORR of patients with high CD8/PD-L1 co-expression group (n=39) was significantly higher than that of (n=38) in the low co-expression group (71.8% vs 47.4%, P = 0.029; Figure 2-B); while in the chemotherapy group, the ORR of the CD8/PD-L1 high and low co-expression group was similar to (P 0.05; Figure 2-B).
Note: The percentage of positive cells in the high co-expression group was ≥median level of the population being analyzed; the percentage of positive cells in the low co-expression group was ≥median level of the population being analyzed.
Figure 2-B Comparison between treatment responses between high and low levels of CD8/PD-L1 co-expression patients
In the carrilizumab combined with chemotherapy group, patients with high CD8/PD-L1 co-expression group had significantly prolonged patients with PFS (P=0.002; Figure 2-C) and OS (P=0.006; Figure 2-D) and OS (P=0.006; Figure 2-D) were significantly prolonged in patients with lower CD8/PD-L1 co-expression group. In the high CD8/PD-L1 co-expression group, PFS (P=0.003) and OS (P=0.032) in patients treated with carilizumab combined with chemotherapy were significantly prolonged compared with those in the chemotherapy group. In the low CD8/PD-L1 co-expression group, there was no significant difference in PFS and OS among patients receiving different treatment methods (Figure 2 C-D) .
Figure 2 Comparison of PFS and OS in patients with high and low CD8/PD-L1 co-expression levels
CD68/PD-L1 co-expression group
3 (n=39) patients with high CD68/PD-L1 co-expression (69.2% vs 50.0%, P=0.085; Figure 3-B) , but did not reach statistical significance; in the chemotherapy group, the ORR of CD68/PD-L1 high and low co-expression group was similar to (P0.05; Figure 3-B) .
Figure 3-B Comparison between treatment responses between patients with high and low levels of CD68/PD-L1 coexpression
3 in the carrilizumab combined with chemotherapy group, patients with high CD68/PD-L1 coexpression were significantly prolonged. In the high CD68/PD-L1 co-expression group, PFS (P=0.004; Figure 3-C) and OS (P=0.022; Figure 3-D) were also significantly better than those treated with chemotherapy alone; while in the low CD68/PD-L1 co-expression group, there was no significant difference in PFS and OS in patients receiving different treatment modes (Figure 3 C-D) .
Figure 3 Comparison of PFS and OS in patients with high and low CD68/PD-L1 co-expression levels
In addition, after adjusting the clinical pathological characteristics, the predictive value of CD8/PD-L1 and CD68/PD-L1 co-expression on the PFS and OS in the carrilizumab combined with chemotherapy group is still statistically significant. The proportion of TP53/KRAS co-mutation in the
CD8/PD-L1 or CD68/PD-L1 high co-expression group was relatively high, and rich PI3K and cell cycle pathway mutations were found in the CD8/PD-L1 co-expression positive group.
Finally, this study evaluated tumor mutation characteristics at different co-expression levels of CD8/PD-L1 or CD68/PD-L1. The results showed that the incidence of TP53 or KRAS mutations was similar between patients with high and low co-expression groups of CD8/PD-L1 or CD68/PD-L1, but the proportion of TP53/KRAS co-mutation in the high co-expression group of CD8/PD-L1 or CD68/PD-L1 was significantly higher than that of in the low co-expression group (both 13.0% vs 0.0%, P=0.023; Figure 4-A) . The CNV analysis results showed that chromosomes 5p and 8q were significantly deletions in the CD8/PD-L1 positive group, and chromosomes 5p, 7p and 14q were locally amplified in the CD8/PD-L1 negative group (Figure 4-B) , and the CD68/PD-L1 positive group and negative group were significantly observed on chromosomes 8q, 10q, 12q and 14q. In addition, compared with the negative group, significantly rich PI3K (P=0.012; Figure 4-C) and cell cycle pathway mutations (P=0.021; Figure 4-D) were found in the CD8/PD-L1 positive group, while there was no difference between the two pathways between the CD68/PD-L1 positive group and the negative group (P>0.05).
Figure 4 Tumor mutation characteristics in the positive and negative group of CD8/PD-L1 co-expression
small 3 link
This study mainly explored the predictive value of the expression levels of tumor immune microenvironment biomarkers PD-L1, CD8, CD68, CD4, FOXP3 and their different combinations in locally advanced or metastatic NSCLC patients treated with PD-1 inhibitor combined with chemotherapy. The results of the
study showed that the CD8/PD-L1 or CD68/PD-L1 co-expression levels in patients with
responded to carilizumab combined with chemotherapy were significantly higher.
In the carrilizumab combined with chemotherapy group, high co-expression of CD8/PD-L1 or CD68/PD-L1 was significantly correlated with PFS and OS prolongation. Patients with high co-expression of
CD8/PD-L1 or CD68/PD-L1 had significantly higher co-mutation ratios of TP53/KRAS, and patients with positive CD8/PD-L1 co-expression of mutations were more abundant in PI3K and cell cycle pathways.
In summary, the biological rational combination of tumor immune microenvironment markers, especially the co-expression of CD8/PD-L1 and CD68/PD-L1, may become the predictive biomarker , a first-line treatment of PD-1 inhibitor combined with chemotherapy in patients with locally advanced or metastatic NSCLC. The relevant predictive value needs to be further verified by subsequent studies.
