The Chromosome 11q13.3 Amplification Induces Hyperprogression To PD-1 Blockade Via PD-1~+Treg Cells

Background and ObjectionA revolutionary cancer therapy targeting the inhibitory immune-checkpoint receptors,such as monoclonal antibodies against programmed cell death 1(PD-1)and PD-1 ligand(PD-L1),have been introduced as a breakthrough in the treatment of a large number of cancer types and dramatically modified the established treatment modality for several advanced cancers.However,more than half of the patients still failed to benefit from Immune checkpoint inhibitors(ICIs),and 4-29%patients even had the "hyperprogressive"(Hyperprogression Disease,HPD)phenomenon that the tumor did not shrink but accelerated abnormally after ICIs treatment.HPD is connected with an inferior outcome and seriously affects the clinical application of immunotherapy.Previous clinical cases reported that the amplification of MDM2/MDM4,EGFR or CCND1/FGF3/FGF4/FGF19 genes in chromosome 11q13.3 may be related to hyperprogression after immunotherapy.However,these findings were not confirmed by reliable experimental evidence,and need further exploration of effective solution to treat patients with HPD.Therefore,it is critical to develop biomarker and therapeutic strategies for patients with drug resistance of tumor immunotherapy,especially with HPD.MethodsTo analyze the prognostic impact of the amplification of CCND1/FGF3/FGF4/FGF19 in chromosome 11q13.3 to immunotherapy-treated patients,we performed whole exon sequencing(WES)of the intrinsic resistant case with accelerated progression to anti-PD-1 monoclonal antibody(Pembrolizumab)and integrated analysis of the correlation between copy number variation(CNV)and clinical immunotherapeutic efficacy in clinical database with public clinical trial.Meanwhile,two murine cancer cell lines with high immunogenicity,Hepa1-6 and MC38,were retroviral transfected and then subcutaneously injected in immunocompetent mice to establish mouse model in response to anti-PD-1 monoclonal antibody.In order to in-depth investigate the molecular mechanism of HPD,co-culture system in vitro and multiple subcutaneous murine models bearing Heap1-6/co-Amp tumors,including immunodeficient mice,Treg cells-depleted in syngeneic C57BL/6 mice by anti-CD25 mAb and CD4+T cells deficiency syngeneic C57BL/6 mice(CD4-/-mice),were established.Then using flow cytometry analysis to exam whether CCND1/FGF3/FGF4/FGF19 co-amplification have an impact on tumor immune microenvironment,and performing transcriptome analysis to determine the molecular mechanism of hyperprogression induced by CCND1/FGF3/FGF4/FGF19 co-amplification.ResultsIt was found that co-amplification of CCND1/FGF3/FGF4/FGF19 in chromosome 11q13.3 is significantly deteriorated the prognosis of immunotherapy-treated patients across many types of cancers and induces HPD to anti-PD-1 antibody in murine models.We found increased proportion of Treg cells and terminally exhausted T cells,decreased infiltration and activity of cytotoxic lymphocytes in the co-Amp tumors.Anti-PD-1 antibody treatment further increased the proportion of PD-1+ Treg cells,which exhibit higher proliferative and immunosuppressive activity,in tumor infiltrating lymphocytes.And the lack of Treg cells completely abrogated the hyperprogression in co-Amp tumors sustained by anti-PD-1 mAb.We preliminary confirmed that the up-regulated amino acids transporter SLC7A5 on tumor cells,which may be responsible for tumor cells avidly consumed amino acids and outcompeted cytotoxic T cells for amino acids,and the reduced pro-inflammatory cytokines and chemokines such as IL6,CCL2,CXCL11 and CXCL14,were regulated by Wnt/β-catenin/c-Myc signaling pathway.The pan-FGFR inhibitor erdafitinib act synergistically with anti-PD-1 mAb on improving anti-tumor response in vitro,and combination therapy also led to inhibition of tumor growth and a survival advantage in CCND1/FGF3/FGF4/FGF19 co-amplification tumor cells-bearing mice by restore the PD-1+ Treg-mediated immunosuppressive tumor microenvironment.In addition,combined therapy significantly rescued the expression of IL-6,CCL2,CXCL11 and CXCL14 and suppressed SLC7A5 expression in Hepal-6/co-Amp tumors,consistent with the synergistic effect on reversing the immunosuppressive TME.Conclusions(1)Co-amplification of CCND1/FGF3/FGF4/FGF19 due to chromosome 11q13.3 amplification induced tumor hyperprogression to anti-PD-1 therapy.(2)The hyperprogression in co-Amp tumors sustained by anti-PD-1 mAb,most likely dependent on PD-1+ Treg cells in tumor microenvironment.(3)CCND1/FGF3/FGF4/FGF15 co-amplification promotes the expression of SLC7A5,outcompeting cytotoxic T cells for amino acids,and reduces the expression of IL-6,CCL2,CXCL11 and CXCL14 to develop immune-suppressive TME via Wnt/β-catenin signaling.(4)Combination therapy using pan-FGFR inhibitor and anti-PD-1 mAb might be useful to overcame 11q13.3 amplification-induced HPD.