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pCDCAR1 GPC3 h(28ζ) (CAR-ZP126)


All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

The vector of anti-GPC3 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target Human GPC3. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-GPC3 antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of HCC.

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Details

  • Target
  • GPC3
  • Targeting Cell Type
  • NK cell
  • Targeting Diseases
  • HCC
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Lentiviral vector
  • Receptor Construction
  • scFv-CD28-CD3ζ
  • Discription of Signaling Cassetes
  • CD28
    CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatory signals required for T cell activation and survival. CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2) proteins which are expressed on antigen-presenting cells (APC). CD28 modulates the primary TCR/CD3ζ signal in a different fashion than the late costimulatory elements OX40 and 4-1BB. CD28 enhances the expression of downstream regulators that impact on T-cell proliferation, death, differentiation, and effector functions. CAR T cell containing the CD28 endodomain showed strikingly enhanced sustained T cell activation, growth, survival. And CD28 results in a brightly expressed, stable receptor as the transmembrane domain. Including CD28 costimulatory domains in CARs led to enhanced anti-malignancy efficacy.
    CD3ζ
    CD3ζ, also known as T-cell receptor zeta, which together with T-cell receptor and CD3γ, δ, ε chain, forms the TCR-CD3 complex. ζ was expressed independently from the complex. The zeta chain plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. CD3-zeta, which contains 3 ITAMs, is the most commonly used endodomain component of CARs. It transmits an activation signal to the T cell after antigen is bound. CD3-zeta may not provide a fully competent activation signal and additional co-stimulatory signaling is needed. For example, chimeric CD28 and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.

Target

  • Clone
  • hu9F2
  • Host
  • Humanized
  • Target Species
  • Human
  • Gene Name
  • glypican 3
  • Synonyms
  • GPC3;GPC3; glypican 3; SGB; DGSX; MXR7; SDYS; SGBS; OCI-5; SGBS1; GTR2-2;

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  • Published Data
Complete CAR data FCM

Fig.1 Surface expression of sFv-ε and CARs on primary human CART cells.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.1 Surface expression of sFv-ε and CARs on primary human CART cells.

Flow cytometry analysis of surface expression of sFv-ε and CARs on primary human CART cells.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data FCM

Fig.2 Surface expression of CD25 and CD69 of UTD or 9F2-CART cells.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.2 Surface expression of CD25 and CD69 of UTD or 9F2-CART cells.

Flow cytometry analysis of surface expression of CD25 and CD69 of UTD or 9F2-CART cells.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.3 Surface expression of CD25 and CD69 of UTD or 9F2-CART cells.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.3 Surface expression of CD25 and CD69 of UTD or 9F2-CART cells.

Figure shows MFI of Flow cytometry analysis of surface expression of CD25 and CD69 of UTD or 9F2-CART cells.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.4 Frequencies of CD45RA+ CCR7+ naive (TN) and CD45RA+ CCR7- terminally differentiated effector memory T cell (TEMRA) subsets in CD8+ T cells.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.4 Frequencies of CD45RA+ CCR7+ naive (TN) and CD45RA+ CCR7- terminally differentiated effector memory T cell (TEMRA) subsets in CD8+ T cells.

Flow cytometry analysis of CD45RA+ CCR7+ naive (TN) and CD45RA+ CCR7- terminally differentiated effector memory T cell (TEMRA) subsets in CD8+ T cells.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.5 Frequencies of CD45RA+ CCR7+ naive (TN) and CD45RA+ CCR7- terminally differentiated effector memory T cell (TEMRA) subsets in CD8+ T cells.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.5 Frequencies of CD45RA+ CCR7+ naive (TN) and CD45RA+ CCR7- terminally differentiated effector memory T cell (TEMRA) subsets in CD8+ T cells.

Figure shows percentage of CD45RA+ CCR7+ naive (TN) and CD45RA+ CCR7- terminally differentiated effector memory T cell (TEMRA) subsets in CD8+ T cells.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data FCM

Fig.6 Surface expression of PD-1 and LAG-3 in UTD or 9F2 CART cells

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.6 Surface expression of PD-1 and LAG-3 in UTD or 9F2 CART cells

Flow cytometry analysis of surface expression of PD-1 and LAG-3 in UTD or 9F2 CART cells

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data FCM

Fig.7 Surface expression of PD-1 and LAG-3 in UTD or 9F2 CART cells

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.7 Surface expression of PD-1 and LAG-3 in UTD or 9F2 CART cells

Figure shows percentage of surface expression of PD-1 and LAG-3 in UTD or 9F2 CART cells

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.8 Characterization of antitumor activity of human 28z CAR, BBz CAR, and sFv-ε T cells

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.8 Characterization of antitumor activity of human 28z CAR, BBz CAR, and sFv-ε T cells

T cells were co-incubated with SK-Hep-1 or PLC/PRF/5 tumor cells at the indicated effector to target (E:T) ratios for 18 h (n = 3-4). Cytotoxic activities were measured using a standard nonradioactive cytotoxicity assay.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.9 Characterization of cytokine production of human 28z CAR, BBz CAR, and sFv-ε T cells

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.9 Characterization of cytokine production of human 28z CAR, BBz CAR, and sFv-ε T cells

T cells were cocultured with PLC/PRF/5 cells with or without monocytes at the ratio of 1:1:1 or 1:1:0 for 48 h. IL6, IFNg, TNFa, and IL2 in co-culture supernatants were quantified by cytometric bead array (CBA) assays, and GM-CSF was examined by ELISA (n = 3).

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.10 CD3+CD8+ and CD3+ CD4+ T cells in peripheral blood were measured with Trucount tubes 7 days after treatment (n = 3).

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.10 CD3+CD8+ and CD3+ CD4+ T cells in peripheral blood were measured with Trucount tubes 7 days after treatment (n = 3).

PLC/PRF/5 tumor cells subcutaneously injected into NPG mice on day 0. On day 13, mice bearing tumors were treated with a single intravenous infusion of CART cells.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.11 Tumor Growth analysis after treatment.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.11 Tumor Growth analysis after treatment.

Growth of PLC/PRF/5 xenografts were monitored over 29 days (n = 5).

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.12 Tumor weights analysis at the end of the experiment.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.12 Tumor weights analysis at the end of the experiment.

Tumor weights at the end of the experiment (n = 5).

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.13 Characterization of cytokine production of human 28z CAR, BBz CAR, and sFv-ε T cells

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.13 Characterization of cytokine production of human 28z CAR, BBz CAR, and sFv-ε T cells

Tumor-bearing mice were intravenously infused with 1.5e7 9F2 CAR-T or UTD T cells. Human IFNg, TNFa, IL2, and murine IL6 in peripheral blood were measured 3 and 6 days after treatment (n = 6 from 3 different mice of each group). Cytokines were also measured in tumor tissues harvested 6 days after treatment (n = 3).

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.14 Characterization of cytokine production of human 28z CAR, BBz CAR, and sFv-ε T cells stimulated with GPC3 peptide.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.14 Characterization of cytokine production of human 28z CAR, BBz CAR, and sFv-ε T cells stimulated with GPC3 peptide.

T cells were stimulated with GPC3 peptide of gradient concentrations for 8 h, and the cytokines in supernatants were quantified.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data Funcs

Fig.15 Characterization of cytokine production of human 28z CAR, BBz CAR, and sFv-ε T cells co-incubated with PLC/PRF/5 cells.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.15 Characterization of cytokine production of human 28z CAR, BBz CAR, and sFv-ε T cells co-incubated with PLC/PRF/5 cells.

T cells were co-incubated with PLC/PRF/5 cells at a 1:1 ratio with or without calcineurin inhibitor FK506 (0.25 nM) for 8 h, and the cytokines in supernatants were measured (n = 3).

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

Complete CAR data FCM

Fig.16 Flow cytometry analysis of intracellular calcium level of human 28z CAR, BBz CAR, and sFv-ε T cells co-incubated with PLC/PRF/5 cells.

CAR Construction : 9F2 scfv-CD3ζ Latest CAR Construction

Fig.16 Flow cytometry analysis of intracellular calcium level of human 28z CAR, BBz CAR, and sFv-ε T cells co-incubated with PLC/PRF/5 cells.

The intracellular calcium level of sFv-ε or CAR T cells stimulated with PLC/PRF/5 cells were analyzed by flow cytometry.

Sun, Y., Dong, Y., Sun, R., Liu, Y., Wang, Y., Luo, H., ... & Li, Z. (2022). Chimeric anti-GPC3 sFv-CD3ε receptor-modified T cells with IL7 co-expression for the treatment of solid tumors. Molecular Therapy-Oncolytics, 25, 160-173.

More Published Data More Published Data

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