Cbl-b inhibitor NX-1607 activates MAPK/ERK signaling pathway and enhances T-cell activation
Backgroud: The E3 ubiquitin ligase casitas B lymphoma-b (Cbl-b) is pivotal in modulating immune responses by attenuating T-cell activation and cytokine production. Inhibiting Cbl-b presents a potential therapeutic strategy in immuno-oncology by enhancing immune activity.Methods: A rapid Homogeneous...
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| Format: | Article |
| Language: | English |
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BMJ Publishing Group
2025-05-01
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| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/13/5/e011180.full |
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| author | Ting Yu Li Jia Wenting Zhu Shan Lu Yongliang Zhang Benjin Liu Shanshan Song Xubin Bao Zehong Miao Jinxue He |
| author_facet | Ting Yu Li Jia Wenting Zhu Shan Lu Yongliang Zhang Benjin Liu Shanshan Song Xubin Bao Zehong Miao Jinxue He |
| author_sort | Ting Yu |
| collection | DOAJ |
| description | Backgroud: The E3 ubiquitin ligase casitas B lymphoma-b (Cbl-b) is pivotal in modulating immune responses by attenuating T-cell activation and cytokine production. Inhibiting Cbl-b presents a potential therapeutic strategy in immuno-oncology by enhancing immune activity.Methods: A rapid Homogeneous Time-Resolved Fluorescence (HTRF) assay was employed to evaluate the inhibitory efficacy of NX-1607 on Cbl-b. The effects of NX-1607 on T cell activation, cytokine production, and proliferation were characterized invitro using primary T cells and Jurkat T cells. A drug combination screening was performed utilizing the CD69 marker via flow cytometry to dentify signaling pathways involved in T cell activation by NX-1607. CRISPR/Cas9 technology was used to knock out PLCG1 and MAPK3/1 in Jurkat T cells, followed by the detection of p-PLCγ1 and p-ERK1/2 though Western blotting. The antitumor efficacy of NX-1607 was assessed in a murine model of A20 B-cell lymphoma using BALB/c mice, with subsequent flow cytometry analysis conducted to examine the phenotype of tumor-infiltrating lymphocytes (TILs).Results: Our data show that NX-1607 effectively inhibits Cbl-b activity at low nanomolar levels, boosting PLCγ1 and HCSL1 phosphorylation, activating MAPK/ERK signaling, and elevating CD69 expression. Inhibiting PLCγ1 and ERK1/2 significantly reduces NX-1607’s effect on T-cell activation. Oral administration of NX-1607 notably decreases tumor growth in the A20 B-cell lymphoma model, with immunophenotyping analyses of tumor-infiltrating lymphocytes revealing increased CD3+, CD4+, and CD8+ T cells in treated tumors. Furthermore, our results demonstrate that treatment with NX-1607 results in increased levels of phosphorylated PLCγ1 and ERK1/2 in circulating T cells.Conclusion: Taken together, these findings imply that the inhibition of Cbl-b by NX-1607 may enhance the activation of the MAPK/ERK signaling pathway, thereby sustaining T-cell activation. This provides compelling evidence for the molecular mechanism of NX-1607, underscoring the pivotal role of Cbl-b in controlling signal strength in T-cell activation after T-cell receptor (TCR) engagement. |
| format | Article |
| id | doaj-art-2601f7e3d3504bcc893cf028eab0f0a4 |
| institution | DOAJ |
| issn | 2051-1426 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj-art-2601f7e3d3504bcc893cf028eab0f0a42025-08-20T03:05:57ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262025-05-0113510.1136/jitc-2024-011180Cbl-b inhibitor NX-1607 activates MAPK/ERK signaling pathway and enhances T-cell activationTing Yu0Li Jia1Wenting Zhu2Shan Lu3Yongliang Zhang4Benjin Liu5Shanshan Song6Xubin Bao7Zehong Miao8Jinxue He93 State Key Laboratory of Chemical Biology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China1 State Key Laboratory of Drug Research, Cancer Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China1 State Key Laboratory of Drug Research, Cancer Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China1 State Key Laboratory of Drug Research, Cancer Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China3 State Key Laboratory of Chemical Biology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China1 State Key Laboratory of Drug Research, Cancer Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China1 State Key Laboratory of Drug Research, Cancer Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China1 State Key Laboratory of Drug Research, Cancer Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China1 State Key Laboratory of Drug Research, Cancer Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China1 State Key Laboratory of Drug Research, Cancer Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, ChinaBackgroud: The E3 ubiquitin ligase casitas B lymphoma-b (Cbl-b) is pivotal in modulating immune responses by attenuating T-cell activation and cytokine production. Inhibiting Cbl-b presents a potential therapeutic strategy in immuno-oncology by enhancing immune activity.Methods: A rapid Homogeneous Time-Resolved Fluorescence (HTRF) assay was employed to evaluate the inhibitory efficacy of NX-1607 on Cbl-b. The effects of NX-1607 on T cell activation, cytokine production, and proliferation were characterized invitro using primary T cells and Jurkat T cells. A drug combination screening was performed utilizing the CD69 marker via flow cytometry to dentify signaling pathways involved in T cell activation by NX-1607. CRISPR/Cas9 technology was used to knock out PLCG1 and MAPK3/1 in Jurkat T cells, followed by the detection of p-PLCγ1 and p-ERK1/2 though Western blotting. The antitumor efficacy of NX-1607 was assessed in a murine model of A20 B-cell lymphoma using BALB/c mice, with subsequent flow cytometry analysis conducted to examine the phenotype of tumor-infiltrating lymphocytes (TILs).Results: Our data show that NX-1607 effectively inhibits Cbl-b activity at low nanomolar levels, boosting PLCγ1 and HCSL1 phosphorylation, activating MAPK/ERK signaling, and elevating CD69 expression. Inhibiting PLCγ1 and ERK1/2 significantly reduces NX-1607’s effect on T-cell activation. Oral administration of NX-1607 notably decreases tumor growth in the A20 B-cell lymphoma model, with immunophenotyping analyses of tumor-infiltrating lymphocytes revealing increased CD3+, CD4+, and CD8+ T cells in treated tumors. Furthermore, our results demonstrate that treatment with NX-1607 results in increased levels of phosphorylated PLCγ1 and ERK1/2 in circulating T cells.Conclusion: Taken together, these findings imply that the inhibition of Cbl-b by NX-1607 may enhance the activation of the MAPK/ERK signaling pathway, thereby sustaining T-cell activation. This provides compelling evidence for the molecular mechanism of NX-1607, underscoring the pivotal role of Cbl-b in controlling signal strength in T-cell activation after T-cell receptor (TCR) engagement.https://jitc.bmj.com/content/13/5/e011180.full |
| spellingShingle | Ting Yu Li Jia Wenting Zhu Shan Lu Yongliang Zhang Benjin Liu Shanshan Song Xubin Bao Zehong Miao Jinxue He Cbl-b inhibitor NX-1607 activates MAPK/ERK signaling pathway and enhances T-cell activation Journal for ImmunoTherapy of Cancer |
| title | Cbl-b inhibitor NX-1607 activates MAPK/ERK signaling pathway and enhances T-cell activation |
| title_full | Cbl-b inhibitor NX-1607 activates MAPK/ERK signaling pathway and enhances T-cell activation |
| title_fullStr | Cbl-b inhibitor NX-1607 activates MAPK/ERK signaling pathway and enhances T-cell activation |
| title_full_unstemmed | Cbl-b inhibitor NX-1607 activates MAPK/ERK signaling pathway and enhances T-cell activation |
| title_short | Cbl-b inhibitor NX-1607 activates MAPK/ERK signaling pathway and enhances T-cell activation |
| title_sort | cbl b inhibitor nx 1607 activates mapk erk signaling pathway and enhances t cell activation |
| url | https://jitc.bmj.com/content/13/5/e011180.full |
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