Colocalized targeting of TGF-β and PD-L1 by bintrafusp alfa elicits distinct antitumor responses
Background Bintrafusp alfa (BA) is a bifunctional fusion protein designed for colocalized, simultaneous inhibition of two immunosuppressive pathways, transforming growth factor-β (TGF-β) and programmed death-ligand 1 (PD-L1), within the tumor microenvironment (TME). We hypothesized that targeting PD...
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| Format: | Article |
| Language: | English |
| Published: |
BMJ Publishing Group
2022-07-01
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| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/10/7/e004122.full |
| _version_ | 1828328902437634048 |
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| author | Feng Jiang Hong Wang Yan Lan Guozhong Qin Jin Qi Bo Marelli Huakui Yu Kin-Ming Lo Dong Zhang Hui Huang Hong Ma Tsz-Lun Yeung Ansgar A Wegener Somdutta Saha Mira Toister-Achituv Molly H Jenkins Li-Ya Chiu Adam Lazorchak Ohad Tarcic George Locke Doron Kalimi Alec W Gross Melissa G Derner Maria Soloviev Mathieu Botte Aroop Sircar Vanita D Sood |
| author_facet | Feng Jiang Hong Wang Yan Lan Guozhong Qin Jin Qi Bo Marelli Huakui Yu Kin-Ming Lo Dong Zhang Hui Huang Hong Ma Tsz-Lun Yeung Ansgar A Wegener Somdutta Saha Mira Toister-Achituv Molly H Jenkins Li-Ya Chiu Adam Lazorchak Ohad Tarcic George Locke Doron Kalimi Alec W Gross Melissa G Derner Maria Soloviev Mathieu Botte Aroop Sircar Vanita D Sood |
| author_sort | Feng Jiang |
| collection | DOAJ |
| description | Background Bintrafusp alfa (BA) is a bifunctional fusion protein designed for colocalized, simultaneous inhibition of two immunosuppressive pathways, transforming growth factor-β (TGF-β) and programmed death-ligand 1 (PD-L1), within the tumor microenvironment (TME). We hypothesized that targeting PD-L1 to the tumor by BA colocalizes the TGF-β trap (TGF-βRII) to the TME, enabling it to sequester TGF-β in the tumor more effectively than systemic TGF-β blockade, thereby enhancing antitumor activity.Methods Multiple technologies were used to characterize the TGF-β trap binding avidity. BA versus combinations of anti-PD-L1 and TGF-β trap or the pan-TGF-β antibody fresolimumab were compared in proliferation and two-way mixed lymphocyte reaction assays. Immunophenotyping of tumor-infiltrating lymphocytes (TILs) and RNA sequencing (RNAseq) analysis assessing stromal and immune landscape following BA or the combination therapy were performed in MC38 tumors. TGF-β and PD-L1 co-expression and their associated gene signatures in MC38 tumors and human lung carcinoma tissue were studied with single-cell RNAseq (scRNAseq) and immunostaining. BA-induced internalization, degradation, and depletion of TGF-β were investigated in vitro.Results BA and fresolimumab had comparable intrinsic binding to TGF-β1, but there was an ~80× avidity-based increase in binding affinity with BA. BA inhibited cell proliferation in TGF-β-dependent and PD-L1-expressing cells more potently than TGF-β trap or fresolimumab. Compared with the combination of anti-PD-L1 and TGF-β trap or fresolimumab, BA enhanced T cell activation in vitro and increased TILs in MC38 tumors, which correlated with efficacy. BA induced distinct gene expression in the TME compared with the combination therapy, including upregulation of immune-related gene signatures and reduced activities in TGF-β-regulated pathways, such as epithelial-mesenchymal transition, extracellular matrix deposition, and fibrosis. Regulatory T cells, macrophages, immune cells of myeloid lineage, and fibroblasts were key PD-L1/TGF-β1 co-expressing cells in the TME. scRNAseq analysis suggested BA modulation of the macrophage phenotype, which was confirmed by histological assessment. PD-L1/TGF-β1 co-expression was also seen in human tumors. Finally, BA induced TGF-β1 internalization and degradation in the lysosomes.Conclusion BA more effectively blocks TGF-β by targeting TGF-β trap to the tumor via PD-L1 binding. Such colocalized targeting elicits distinct and superior antitumor responses relative to single agent combination therapy. |
| first_indexed | 2024-04-13T20:16:21Z |
| format | Article |
| id | doaj.art-6670ff6236be4d8e8fdcf07fa04a27b4 |
| institution | Directory Open Access Journal |
| issn | 2051-1426 |
| language | English |
| last_indexed | 2024-04-13T20:16:21Z |
| publishDate | 2022-07-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj.art-6670ff6236be4d8e8fdcf07fa04a27b42022-12-22T02:31:42ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262022-07-0110710.1136/jitc-2021-004122Colocalized targeting of TGF-β and PD-L1 by bintrafusp alfa elicits distinct antitumor responsesFeng Jiang0Hong Wang1Yan Lan2Guozhong Qin3Jin Qi4Bo Marelli5Huakui Yu6Kin-Ming Lo7Dong Zhang8Hui Huang9Hong Ma10Tsz-Lun Yeung11Ansgar A Wegener12Somdutta Saha13Mira Toister-Achituv14Molly H Jenkins15Li-Ya Chiu16Adam Lazorchak17Ohad Tarcic18George Locke19Doron Kalimi20Alec W Gross21Melissa G Derner22Maria Soloviev23Mathieu Botte24Aroop Sircar25Vanita D Sood26Department of Epidemiology, School of Public Health, Fudan University, Shanghai, ChinaDepartment of Anesthesiology, The First Medical Center of Chinese PLA General Hospital, Beijing, ChinaAff2 grid.481568.6EMD Serono Research & Development Institute, Inc. Billerica MA USAAff168 grid.467308.e0000 0004 0412 6436EMD Serono Billerica MA USAAff168 grid.467308.e0000 0004 0412 6436EMD Serono Billerica MA USAAff168 grid.467308.e0000 0004 0412 6436EMD Serono Billerica MA USAAff168 grid.467308.e0000 0004 0412 6436EMD Serono Billerica MA USAAff168 grid.467308.e0000 0004 0412 6436EMD Serono Billerica MA USA4 Department of Neurosurgery, Beijing Tiantan Hospital, Beijing, ChinaDepartment of Ophthalmology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, ChinaDepartment of Cardiovascular Medicine, Key Laboratory on Assisted Circulation, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, ChinaEMD Serono, Billerica, MA USA, Billerica, MA, USADepartment of Discovery and Development Technologies, Merck Healthcare KGaA, Darmstadt, GermanyDepartment of Translational Medicine, EMD Serono Research and Development Institute, Billerica, Massachusetts, USADepartment of Discovery and Development Technologies, Merck Healthcare KGaA, Yavne, IsraelDepartment of TIP OIO, EMD Serono Research and Development Institute, Billerica, Massachusetts, USADepartment of TIP OIO, EMD Serono Research and Development Institute, Billerica, Massachusetts, USADepartment of TIP OIO, EMD Serono Research and Development Institute, Billerica, Massachusetts, USADepartment of Discovery and Development Technologies, Merck Healthcare KGaA, Yavne, IsraelDepartment of Translational Medicine, EMD Serono Research and Development Institute, Billerica, Massachusetts, USADepartment of Discovery and Development Technologies, Merck Healthcare KGaA, Yavne, IsraelDepartment of Discovery Development Technologies, EMD Serono Research and Development Institute, Billerica, Massachusetts, USADepartment of TIP OIO, EMD Serono Research and Development Institute, Billerica, Massachusetts, USADepartment of Discovery Development Technologies, EMD Serono Research and Development Institute, Billerica, Massachusetts, USALeadXPro AG, Villigen, SwitzerlandDepartment of TIP OIO, EMD Serono Research and Development Institute, Billerica, Massachusetts, USADepartment of Discovery Development Technologies, EMD Serono Research and Development Institute, Billerica, Massachusetts, USABackground Bintrafusp alfa (BA) is a bifunctional fusion protein designed for colocalized, simultaneous inhibition of two immunosuppressive pathways, transforming growth factor-β (TGF-β) and programmed death-ligand 1 (PD-L1), within the tumor microenvironment (TME). We hypothesized that targeting PD-L1 to the tumor by BA colocalizes the TGF-β trap (TGF-βRII) to the TME, enabling it to sequester TGF-β in the tumor more effectively than systemic TGF-β blockade, thereby enhancing antitumor activity.Methods Multiple technologies were used to characterize the TGF-β trap binding avidity. BA versus combinations of anti-PD-L1 and TGF-β trap or the pan-TGF-β antibody fresolimumab were compared in proliferation and two-way mixed lymphocyte reaction assays. Immunophenotyping of tumor-infiltrating lymphocytes (TILs) and RNA sequencing (RNAseq) analysis assessing stromal and immune landscape following BA or the combination therapy were performed in MC38 tumors. TGF-β and PD-L1 co-expression and their associated gene signatures in MC38 tumors and human lung carcinoma tissue were studied with single-cell RNAseq (scRNAseq) and immunostaining. BA-induced internalization, degradation, and depletion of TGF-β were investigated in vitro.Results BA and fresolimumab had comparable intrinsic binding to TGF-β1, but there was an ~80× avidity-based increase in binding affinity with BA. BA inhibited cell proliferation in TGF-β-dependent and PD-L1-expressing cells more potently than TGF-β trap or fresolimumab. Compared with the combination of anti-PD-L1 and TGF-β trap or fresolimumab, BA enhanced T cell activation in vitro and increased TILs in MC38 tumors, which correlated with efficacy. BA induced distinct gene expression in the TME compared with the combination therapy, including upregulation of immune-related gene signatures and reduced activities in TGF-β-regulated pathways, such as epithelial-mesenchymal transition, extracellular matrix deposition, and fibrosis. Regulatory T cells, macrophages, immune cells of myeloid lineage, and fibroblasts were key PD-L1/TGF-β1 co-expressing cells in the TME. scRNAseq analysis suggested BA modulation of the macrophage phenotype, which was confirmed by histological assessment. PD-L1/TGF-β1 co-expression was also seen in human tumors. Finally, BA induced TGF-β1 internalization and degradation in the lysosomes.Conclusion BA more effectively blocks TGF-β by targeting TGF-β trap to the tumor via PD-L1 binding. Such colocalized targeting elicits distinct and superior antitumor responses relative to single agent combination therapy.https://jitc.bmj.com/content/10/7/e004122.full |
| spellingShingle | Feng Jiang Hong Wang Yan Lan Guozhong Qin Jin Qi Bo Marelli Huakui Yu Kin-Ming Lo Dong Zhang Hui Huang Hong Ma Tsz-Lun Yeung Ansgar A Wegener Somdutta Saha Mira Toister-Achituv Molly H Jenkins Li-Ya Chiu Adam Lazorchak Ohad Tarcic George Locke Doron Kalimi Alec W Gross Melissa G Derner Maria Soloviev Mathieu Botte Aroop Sircar Vanita D Sood Colocalized targeting of TGF-β and PD-L1 by bintrafusp alfa elicits distinct antitumor responses Journal for ImmunoTherapy of Cancer |
| title | Colocalized targeting of TGF-β and PD-L1 by bintrafusp alfa elicits distinct antitumor responses |
| title_full | Colocalized targeting of TGF-β and PD-L1 by bintrafusp alfa elicits distinct antitumor responses |
| title_fullStr | Colocalized targeting of TGF-β and PD-L1 by bintrafusp alfa elicits distinct antitumor responses |
| title_full_unstemmed | Colocalized targeting of TGF-β and PD-L1 by bintrafusp alfa elicits distinct antitumor responses |
| title_short | Colocalized targeting of TGF-β and PD-L1 by bintrafusp alfa elicits distinct antitumor responses |
| title_sort | colocalized targeting of tgf β and pd l1 by bintrafusp alfa elicits distinct antitumor responses |
| url | https://jitc.bmj.com/content/10/7/e004122.full |
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