Professor Zhou Caicun CameL research: The co-expression of different tumor immune microenvironment markers has the predictive value of potential immune combination therapy

*For medical professionals only

The research results of Zhenhe Technology and the team of Professor Zhou Caicun of Shanghai Pulmonary Hospital affiliated to Tongji University were published in "Cancer Communications", IF:15.
283
.
In this study, Zhen and Landis products were used for multiplex immunofluorescence (mIF) detection
.

Guide

PD-1 inhibitors in combination with chemotherapy have become the new standard of the standard for first-line treatment in patients with locally advanced or metastatic non-small cell lung cancer (NSCLC), but nearly half of patients still do not benefit
from this combination therapy.
To further expand the population benefiting from PD-1 inhibitors in combination with chemotherapy, it is important
to find reliable predictive biomarkers.
However, neither PD-L1 expression nor single biomarkers such as TMB predicted the efficacy of
combination therapy.
Therefore, by detecting the expression level of tumor immune microenvironment biomarkers, this study explored the predictive value
of a reasonable combination of different markers on the efficacy of PD-1 inhibitors combined with chemotherapy in patients with advanced NSCLC.

1

study

Overview

Patient condition

A total of 144 patients with stage III.
B-IV.
non-squamous NSCLC who were newly treated were included, of which 77 received the PD-1 inhibitor carrelizumab in combination with chemotherapy and 67 received chemotherapy alone, and the objective response rate (ORR) of carrelizumab combined with chemotherapy was significantly higher than that in the chemotherapy alone group (59.
7% vs 37.
3%, P=0.
007).

Sample testing

Multiplex immunofluorescence (mIF) based on multiplex immunofluorescence (mIF) technology was used to detect and analyze
immunomicroenvironmental biomarkers (including PDL1, CD8, CD68, CD4 and FOXP3) in tissue samples prior to treatment.

Table 1 Clinical baseline characteristics of 144 patients with NSCLC

2

The essential

Study results

This study first evaluated the correlation
between expression levels (positive cell density and percentage of positive cells) and objective responses of single and co-expression markers (positive cell density and percentage of positive cells) in responders (CR+PR patients) and non-responders (SD+PD patients) in the carrelizumab plus chemotherapy group.

Single markers (PD-L1, CD8, CD68, CD4, FOXP3)

The PD-L1-positive cell density (P=0.
013), the percentage of positive cells (P=0.
020), CD8-positive cell density (P=0.
046) and CD68-positive cell percentage (P=0.
048) of the responders were significantly higher than those of the nonresponders
.
The expression levels of CD4 and FOXP3 were not correlated with objective response (Figure 1-B).

Figure 1-B Correlation between single marker expression levels and therapeutic efficacy

Co-expression markers (CD8/PD-L1, CD68/PD-L1, CD4/FOXP3, CD8/CD68/PD-L1, CD4/FOXP3/PD-L1)

The positive cell density and percentage of positive cells in the responders were significantly higher than those in the nonresponders, while the co-expression levels of CD4/FOXP3, CD8/CD68/PD-L1, and CD4/FOXP3/PD-L1 were similar
between the responders and nonresponders.

Figure 1-C Correlation between expression levels of co-expression markers and therapeutic efficacy

In the carrelizumab combined with chemotherapy group, the ORR of patients with high coexpression levels of CD8/PD-L1 or CD68/PD-L1 was significantly higher.
The PFS and OS of patients with high co-expression of CD8/PD-L1 and CD68/PD-L1 were significantly improved
.

Next, this study evaluated the correlation
between different combinations of co-expression markers and clinical outcomes in the carrelizumab plus chemotherapy group.
The results showed that high CD8/PD-L1 and high CD68/PD-L1 co-expression levels had better predictive value
.

CD8/PD-L1 co-expression group

In the carrelizumab combined with chemotherapy group, the ORR of patients in the high CD8/PD-L1 co-expression group (n=39) was significantly higher than that in the low co-expression group (n=38) (71.
8% vs 47.
4%, P = 0.
029; Figure 2-B).
In the chemotherapy alone group, the ORR of CD8/PD-L1 high and low co-expression groups was similar (P > 0.
05; Figure 2-B).

Note: The high co-expression group is the percentage of positive cells ≥ median level of the analyzed population; The low co-expression group is the percentage < median level of positive cells in the population being analyzed<b10>.

Figure 2-B Comparison between treatment response in patients with high and low levels of CD8/PD-L1 co-expression

In the carrelizumab combined with chemotherapy group, PFS (P=0.
002; Figure 2-C) and OS (P=0.
006; Figure 2-D) were significantly prolonged
in the high CD8/PD-L1 co-expression group.

In the high CD8/PD-L1 co-expression group, PFS (P=0.
003) and OS (P=0.
032) were significantly longer
in patients receiving carrelizumab combined with chemotherapy than those in the chemotherapy alone.
In the low CD8/PD-L1 co-expression group, there was no significant difference in PFS and OS among patients receiving different treatment modalities (Figure 2 C-D).

Fig.
2 Comparison of PFS and OS in patients with high and low CD8/PD-L1 co-expression levels in C-D

CD68/PD-L1 co-expression group

Patients with high CD68/PD-L1 co-expression (n=39) also achieved better ORR (69.
2% vs 50.
0%, P=0.
085; Figure 3-B) in the carrelizumab combined with chemotherapy group, but did not achieve statistical significance.
In the chemotherapy alone group, the ORR of CD68/PD-L1 high and low co-expression was similar (P>0.
05; Figure 3-B).

Figure 3-B Comparison of treatment responses between patients with high and low CD68/PD-L1 co-expression

In the carrelizumab combined with chemotherapy group, PFS (P=0.
024; Figure 3-C) and OS (P=0.
026; Figure 3-D) and low CD68/PD-L1 co-expression patients with high CD68/PD-L1 co-expression 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) of patients receiving carrelizumab combined with chemotherapy were also significantly better than those receiving chemotherapy alone.
In the low CD68/PD-L1 co-expression group, there was no significant difference in PFS and OS among patients receiving different treatment modalities (Figure 3 C-D).

Fig.
3 Comparison of PFS and OS in patients with high and low CD68/PD-L1 co-expression levels in C-D

In addition, after adjusting the clinicopathological features, the predictive value of CD8/PD-L1 and CD68/PD-L1 co-expression in the carrelizumab combined with chemotherapy group PFS and OS was still statistically significant
.

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 among patients with high and low CD8/PD-L1 or CD68/PD-L1 co-expression groups, but the TP53/KRAS co-mutation ratio in the CD8/PD-L1 or CD68/PD-L1 high co-expression group was significantly higher than that in the low co-expression group (both 13.
0% vs 0.
0%, P=0.
023; Figure 4-A).

The results of CNV analysis showed significant deletion of CD8/PD-L1 positive chromosomes 5p and 8q, local amplification of CD8/PD-L1-negative chromosomes 5p, 7p and 14q (Figure 4-B), and significant CNV
was observed on chromosomes 8q, 10q, 12q and 14q in the CD68/PD-L1 positive and negative groups.

In addition, compared with the negative group, a significant abundant 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 CD68/PD-L1 positive and negative groups (P>0.
05).

Fig.
4 Tumor mutation characteristics of CD8/PD-L1 co-expression positive and negative groups

3

small

knot

This study mainly explores the predictive value of the expression levels of tumor immune microenvironment biomarkers PD-L1, CD8, CD68, CD4, FOXP3 and their different combinations in patients with locally advanced or metastatic NSCLC treated with PD-1 inhibitors and chemotherapy
.

• Patients who responded to carrelizumab combined with chemotherapy had significantly higher
  levels of CD8/PD-L1 or CD68/PD-L1 co-expression.
  

• In the carrelizumab plus chemotherapy group, high co-expression of CD8/PD-L1 or CD68/PD-L1 was significantly associated
  with PFS and OS prolongation.
  

• Patients with high CD8/PD-L1 or CD68/PD-L1 co-expression had significantly higher TP53/KRAS co-mutation ratios, and patients with positive CD8/PD-L1 co-expression had more abundant
  mutations in PI3K and cell cycle pathways.
  

Original research results:

Multiplexed imaging of tumor immune microenvironmental markers in locally advanced or metastatic non-small-cell lung cancer characterizes the features of response to PD-1 blockade plus chemotherapy; 

doi:10.
1002/cac2.
12383

Note: All images in the text are from Wu F y et al .
Cancer Commun (Lond).
2022 Nov 4.