Early Change in the Plasma Levels of Circulating Soluble Immune Checkpoint Proteins in Patients with Unresectable Hepatocellular Carcinoma Treated by Lenvatinib or Transcatheter Arterial Chemoembolization

Naoshi Odagiri, Hoang Hai, Le Thi Thanh Thuy, Minh Phuong Dong, Maito Suoh, Kohei Kotani, Atsushi Hagihara, Sawako Uchida-Kobayashi, Akihiro Tamori, Masaru Enomoto, Norifumi Kawada, Naoshi Odagiri, Hoang Hai, Le Thi Thanh Thuy, Minh Phuong Dong, Maito Suoh, Kohei Kotani, Atsushi Hagihara, Sawako Uchida-Kobayashi, Akihiro Tamori, Masaru Enomoto, Norifumi Kawada

Abstract

Immune checkpoint inhibitors, combined with anti-angiogenic agents or locoregional treatments (e.g., transarterial chemoembolization (TACE)), are expected to become standard-of-care for unresectable hepatocellular carcinoma (HCC). We measured the plasma levels of 16 soluble checkpoint proteins using multiplexed fluorescent bead-based immunoassays in patients with HCC who underwent lenvatinib (n = 24) or TACE (n = 22) treatment. In lenvatinib-treated patients, plasma levels of sCD27 (soluble cluster of differentiation 27) decreased (p = 0.040) and levels of sCD40 (p = 0.014) and sTIM-3 (p < 0.001) were increased at Week 1, while levels of sCD27 (p < 0.001) were increased significantly at Weeks 2 through 4. At Week 1 of TACE, in addition to sCD27 (p = 0.028), sCD40 (p < 0.001), and sTIM-3 (soluble T-cell immunoglobulin and mucin domain-3) (p < 0.001), levels of sHVEM (soluble herpesvirus entry mediator) (p = 0.003), sTLR-2 (soluble Toll-like receptor 2) (p = 0.009), sCD80 (p = 0.036), sCTLA-4 (soluble cytotoxic T-lymphocyte antigen 4) (p = 0.005), sGITR (soluble glucocorticoid-induced tumor necrosis factor receptor) (p = 0.030), sGITRL (soluble glucocorticoid-induced TNFR-related ligand) (p = 0.090), and sPD-L1 (soluble programmed death-ligand 1) (p = 0.070) also increased. The fold-changes in soluble checkpoint receptors and their ligands, including sCTLA-4 with sCD80/sCD86 and sPD-1 (soluble programmed cell death domain-1) with sPD-L1 were positively correlated in both the lenvatinib and TACE treatment groups. Our results suggest that there are some limited differences in immunomodulatory effects between anti-angiogenic agents and TACE. Further studies from multicenters may help to identify an effective combination therapy.

Keywords: HCC; TACE; liver cancer; molecular-targeted agent; tyrosine kinase inhibitor.

Conflict of interest statement

Norifumi Kawada has received lecture fees from Eisai Co. Ltd.

Figures

Figure A1
Figure A1
Changes in soluble immune checkpoint protein levels after 1 week of lenvatinib treatment. Box plots display the sBTLA, sLAG-3, sPD-1, sPD-L1, sICOS, sCD28, sCD86, sCD80, sTLR-2, sGITR, sGITRL, and sCTLA-4 levels in HCC patients at baseline and Week 1 of treatment. The vertical lengths of the boxes indicate the interquartile ranges and the lines in the boxes suggest the median values, while the error bars show the minimum and maximum values (range). The Wilcoxon signed-rank test was used.
Figure A2
Figure A2
Changes in soluble immune checkpoint protein levels after two to four weeks of lenvatinib treatment. Box plots display the sBTLA, sLAG-3, sPD-1, sPD-L1, sICOS, sCD28, sCD86, sCD80, sTLR-2, sGITR, sGITRL, and sCTLA-4 levels in HCC patients at baseline and Week 2 through 4 of treatment. The vertical lengths of the boxes indicate the interquartile ranges and the lines in the boxes show the median values, while the error bars reveal the minimum and maximum values (range). The Wilcoxon signed-rank test was used.
Figure A3
Figure A3
Changes in soluble immune checkpoint protein levels after 1 week of TACE treatment. Box plots display the sBTLA, sLAG-3, sPD-1, sICOS, sCD28, and sCD86 levels in HCC patients at baseline and Week 1 of treatment. The vertical lengths of the boxes reveal the interquartile ranges and the lines in the boxes show the median values, while the error bars suggest the minimum and maximum values (range). The Wilcoxon signed-rank test was used.
Figure A4
Figure A4
The relationships between fold-changes in soluble immune checkpoint protein levels in plasma at two to four weeks of lenvatinib treatment are seen. (a) A positive correlation was found between the fold-changes of soluble checkpoint receptors and their ligands, including sCTLA-4 with sCD80/sCD86 and sPD-1 with sPD-L1, respectively. (b) The fold-change of sCD27 was positively correlated with those of sCD86, sPD-1, and sPD-L1. Spearman’s rank correlation test was used. A p-value of less than 0.05 was considered to be statistically significant.
Figure 1
Figure 1
Changes in soluble immune checkpoint protein levels in plasma after 1 week of lenvatinib treatment. Box plots show the sCD27, sCD40, sTIM-3, and sHVEM levels in HCC patients at baseline and Week 1 of treatment. The vertical lengths of the boxes indicate the interquartile ranges and the lines in the boxes suggest the median values, while the error bars show the minimum and maximum values (in a range). The Wilcoxon signed-rank test was used. HCC, hepatocellular carcinoma; sTIM-3, soluble T-cell immunoglobulin and mucin domain–3; sHVEM, soluble herpesvirus entry mediator; sCD, soluble cluster of differentiation.
Figure 2
Figure 2
Changes in soluble immune checkpoint protein levels in plasma after 2 to 4 weeks of lenvatinib treatment. Box plots demonstrate the sCD27, sCD40, sTIM-3, and sHVEM levels in HCC patients at baseline and Weeks 2 through 4 of treatment. The vertical lengths of the boxes indicate the interquartile ranges and the lines in the boxes display the median values, while the error bars show the minimum and maximum values (in a range). The Wilcoxon signed-rank test was used.
Figure 3
Figure 3
Changes in soluble immune checkpoint protein levels in plasma after 1 week of TACE treatment. Box plots display the sCD27, sCD40, sTIM-3, sHVEM, sTLR-2, sCD80, sCTLA-4, sGITR, sGITRL, and sPD-L1 levels in HCC patients at baseline and Week 1 of treatment. The vertical lengths of the boxes indicate the interquartile ranges and the lines in the boxes suggest the median values, while the error bars show the minimum and maximum values (range). The Wilcoxon signed-rank test was used. TACE, transarterial chemoembolization; sTLR-2, soluble Toll-like receptor 2; sCTLA-4, soluble cytotoxic T-lymphocyte antigen 4; sGITR, soluble glucocorticoid-induced tumor necrosis factor receptor; sGITRL, soluble glucocorticoid-induced TNFR-related ligand; sPD-L1, soluble programmed death-ligand 1.
Figure 4
Figure 4
The relationships between fold-changes in soluble immune checkpoint protein levels in plasma at 1 week of lenvatinib treatment are presented. (a) A positive correlation was found between fold-changes in soluble checkpoint receptors and their ligands, including sCTLA-4 with sCD80/sCD86 and sPD-1 with sPD-L1, respectively. (b) sCD40 was positively correlated with some proteins in fold-changes, including sCD86, sPD-1, and sPD-L1. Spearman’s rank correlation test was used. A p-value of less than 0.05 was considered to be statistically significant.
Figure 5
Figure 5
The relationships between fold-changes in soluble immune checkpoint protein levels in plasma at 1 week of receiving TACE are shown. (a) A positive correlation was found between the fold-changes of soluble checkpoint receptors and their ligands, including sCTLA-4 with sCD80/sCD86 and sPD-1 with sPD-L1. (b) Separately, a strong correlation was observed between the fold-changes in sCD80 and sTLR-2 levels, sCD40 and sHVEM levels, and sHVEM and sTIM-3 levels. Spearman’s rank correlation test was used. A p-value of less than 0.05 was considered to be statistically significant.

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