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pCDCAR1 GD2 h(BBζ) (CAR-MZ046)


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

The vector of anti-GD2 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human GD2. The T cells are genetically modified through transduction with a retroviral vector expressing scFv of anti-GD2 antibody linked to 41BB and TCR ζ chain signaling domains. And the vector product was designed for the treatment of GD2-positive tumors.

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Details

  • Target
  • GD2
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • GD2-positive tumors
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Retroviral
  • Receptor Construction
  • scFv-41BB-CD3ζ
  • Discription of Signaling Cassetes
  • 41BB
    CD137 (also known as 4-1BB) is a surface co-stimulatory glycoprotein originally described as present on activated T lymphocytes, which belongs to the tumor necrosis factor (TNF) receptor superfamily. It is expressed mainly on activated CD4+ and CD8+ T cells, and binds to a high-affinity ligand (4-1BBL) expressed on several antigen-presenting cells such as macrophages and activated B cells. On the basis of preclinical observation, this molecule can promote the persistence of antigen-specific and antigen-nonspecific chimeric antigen receptor T-cells to significantly increases antitumor activity.
    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
  • 14g2a
  • Host
  • Mouse
  • Target Species
  • Human
  • Gene Name
  • Graves disease, susceptibility to, 2
  • Synonyms
  • GD2, GRD2

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  • Published Data
CAR scFv data FCM

FIG.1 Relative affinity and specificity of 14.G2a scFv variants toward GD2.

CAR Construction : Latest CAR Construction

FIG.1 Relative affinity and specificity of 14.G2a scFv variants toward GD2.

Representative histograms showing binding of WT and HGlu101Lys variants of scFv (5 g/ml) to tested neuroblastoma cells (dark gray) overlaid with
control staining of the respective cells with FITC-conjugated anti-His tag antibody alone (light gray).

Horwacik, I., Golik, P., Grudnik, P., Kolinski, M., Zdzalik, M., Rokita, H., & Dubin, G. (2015). Structural basis of GD2 ganglioside and mimetic peptide recognition by 14G2a antibody. Molecular & Cellular Proteomics, 14(10), 2577-2590.

CAR scFv data FCM

FIG.2 Relative affinity and specificity of 14.G2a scFv variants toward GD2.

CAR Construction : Latest CAR Construction

FIG.2 Relative affinity and specificity of 14.G2a scFv variants toward GD2.

MFI signals for single cell populations stained with particular scFv and FITC-conjugated anti-His tag antibody, or the latter antibody alone (control).

Horwacik, I., Golik, P., Grudnik, P., Kolinski, M., Zdzalik, M., Rokita, H., & Dubin, G. (2015). Structural basis of GD2 ganglioside and mimetic peptide recognition by 14G2a antibody. Molecular & Cellular Proteomics, 14(10), 2577-2590.

Complete CAR data FuncS

Fig.3 Cell viability calculation of Glioblastoma cell lines (dsRED T98G GD2high, dsRED A172 GD2low) co-cultures at 2:1 E:T ratio with GD2 CAR T and GFP control T cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.3 Cell viability calculation of Glioblastoma cell lines (dsRED T98G GD2high, dsRED A172 GD2low) co-cultures at 2:1 E:T ratio with GD2 CAR T and GFP control T cells.

Co-cultures at 2:1 E:T ratio with GD2 CAR T. After 48 h tumor cell viability is calculated by fluorescence viability assay.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.4 Cell viability calculation of patient-derived primary glioblastoma cells (dsRED C3, dsRED C6, dsRED C10, and dsRED C12) co-cultures at 2:1 E:T ratio with GD2 CAR T and GFP control T cells derived from five PBMC donors.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.4 Cell viability calculation of patient-derived primary glioblastoma cells (dsRED C3, dsRED C6, dsRED C10, and dsRED C12) co-cultures at 2:1 E:T ratio with GD2 CAR T and GFP control T cells derived from five PBMC donors.

Co-cultures at 2:1 E:T ratio with GD2 CAR T. After 48 h tumor cell viability is calculated by fluorescence viability assay.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.5 GD2 CAR T cells exert specific killing in a time- and dose-dependent manner against several GD2high patient-derived primary glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.5 GD2 CAR T cells exert specific killing in a time- and dose-dependent manner against several GD2high patient-derived primary glioblastoma cells.

Glioblastoma cells (dsRED C3, dsRED C6, dsRED C10, and dsRED C12) co-cultures with GD2 CAR T and GFP control T cells at three different E:T ratios. After 24, 48, and 72 h tumor viability is calculated by fluorescence viability assay.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.6 GD2 CAR T cells generate clusters to eradicate patient-derived primary glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.6 GD2 CAR T cells generate clusters to eradicate patient-derived primary glioblastoma cells.

Representative fluorescence micrographs highlight a robust cluster activation and killing activity of GD2 CAR T population against primary glioblastoma cells.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.7 Autologous GD2 CAR T cells kill glioblastoma in vitro.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.7 Autologous GD2 CAR T cells kill glioblastoma in vitro.

Glioblastoma cells (dsRED C6) co-cultures with either autologous or allogeneic GD2 CAR T and GFP control T cells at 5:1 E:T ratios. After 48 and 72 h tumor viability is calculated by fluorescence assay.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.8 Autologous GD2 CAR T cells kill glioblastoma in vitro.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.8 Autologous GD2 CAR T cells kill glioblastoma in vitro.

Representative fluorescence micrographs show clusters of activation and killing activity of GD2 CAR T cells. GFP T cell population show an absence of reactivity of autologous cytotoxic lymphocytes.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.9 Granzyme release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.9 Granzyme release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 7days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.10 MIP-1α release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.10 MIP-1α release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 8days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.11 IFNγ release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.11 IFNγ release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 9days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.12 TNFα release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.12 TNFα release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 10days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.13 sFAS-Ligand analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.13 sFAS-Ligand analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 11days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.14 TRAIL analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.14 TRAIL analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 12days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.15 GM-CSF release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.15 GM-CSF release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 13days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.16 IL-6 release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.16 IL-6 release analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 14days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.17 CD137 analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.17 CD137 analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 15days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.18 TGF β1 analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.18 TGF β1 analysis during GD2 CAR T cells in co-culture with glioblastoma cells.

Co-culture supernatants of glioblastoma cell lines and primary glioblastoma cells with either GD2 CAR T (light gray) or GFP control T cells (light black) at 2:1 E:T ratio are assessed for cytokine release at 48 h and 16days.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.19 Survival curve of Intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=2:1.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.19 Survival curve of Intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=2:1.

Kaplan-Meier survival curve and log-rank tests are used to measure differences among groups.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.20 Mean total flux values of Intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=2:1.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.20 Mean total flux values of Intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=2:1.

Mean total flux values (photons/s) of either GD2 CAR T or GFP T control-treated mice and control animals monitored by IVIS system. Data are shown as a mean ± SD; p values are calculated by unpaired two-tailed t-test.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.21 Tumor progression monitored with intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=2:1.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.21 Tumor progression monitored with intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=2:1.

Tumor progression is monitored weekly by a luminescence signal with IVIS imaging measuring. DPI day post-injection.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.22 Survival curve of Intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=5:1.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.22 Survival curve of Intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=5:1.

Kaplan-Meier survival curve and log-rank tests are used to measure differences among groups.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.23 Mean total flux values of Intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=5:1.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.23 Mean total flux values of Intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=5:1.

Mean total flux values (photons/s) of either GD2 CAR T or GFP T control-treated mice and control animals monitored by IVIS system.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

Complete CAR data FuncS

Fig.24 Tumor progression monitored with intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=5:1.

CAR Construction : 14.G2a/14g2a-CD3ζ Latest CAR Construction

Fig.24 Tumor progression monitored with intracerebral GD2 CAR T cell treatment in the orthotopic glioblastoma model with E:T=5:1.

Tumor progression is monitored weekly by a luminescence signal with IVIS imaging measuring. DPI day post-injection.

Prapa, M., Chiavelli, C., Golinelli, G., Grisendi, G., Bestagno, M., Di Tinco, R., ... & Dominici, M. (2021). GD2 CAR T cells against human glioblastoma. NPJ precision oncology, 5(1), 1-14.

More Published Data More Published Data

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