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pCDCAR1 MSLN h(BBζ, FKBP/FRB) (CAR-LC270)


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

The vector of anti-mesothelin chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human mesothelin. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-mesothelin antibody, two CD137 (4-1BB) signaling domains, an FKBP and FRB dimerizer-binding pair, and a CD3-zeta signaling domain. The activation of conditional CAR can be controlled by FKBP-FRB dimerization using rapamycin. And the vector product was designed for the treatment of pancreatic cancer, ovarian cancer, lung cancer, malignant pleural mesothelioma tumors (MPM tumors).

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Details

  • Target
  • Mesothelin
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Pancreatic cancer, Ovarian cancer, Lung cancer, Malignant pleural mesothelioma tumors (MPM tumors)
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Lentiviral
  • Receptor Construction
  • scFv-41BB-FKBP/41BB-FRB-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
  • m912
  • Host
  • Human
  • Target Species
  • Human
  • Gene Name
  • mesothelin
  • Synonyms
  • Mesothelin;MSLN; mesothelin; MPF; SMRP;

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  • Published Data
Complete CAR data Cyt

Fig.1 MSLN-directed CAR T cells showed cytotoxic activity in vitro toward human ovarian cancer cell lines expressing MSLN.

CAR Construction : m912 scfv-CD28-CD3ζ; m912 scfv-41BB-CD3ζ Latest CAR Construction

Fig.1 MSLN-directed CAR T cells showed cytotoxic activity in vitro toward human ovarian cancer cell lines expressing MSLN.

Percentage of lysis of parental and MSLN+-transduced ovarian cancer cell lines (OVCAR3 and SKOV3) by M28z and MBBz CAR T cells measured by LDH release assays (n = 4-9).

Schoutrop, E., El-Serafi, I., Poiret, T., Zhao, Y., Gultekin, O., He, R., ... & Mattsson, J. (2021). Mesothelin-Specific CAR T Cells Target Ovarian CancerEvaluation of MSLN CAR T Cells for Ovarian Cancer Treatment. Cancer research, 81(11), 3022-3035.

Complete CAR data Cyt

Fig.2 MSLN-directed CAR T cells showed cytotoxic activity in vitro toward human ovarian cancer cell lines expressing MSLN.

CAR Construction : m912 scfv-CD28-CD3ζ; m912 scfv-41BB-CD3ζ Latest CAR Construction

Fig.2 MSLN-directed CAR T cells showed cytotoxic activity in vitro toward human ovarian cancer cell lines expressing MSLN.

Frequency of CD4+ and CD8+ T cells producing IFNγ, IL2, and TNF (measured by flow cytometry) following exposure of M28z- and MBBz-transduced T cells with MSLN+ SKOV3 cells (n = 7).

Schoutrop, E., El-Serafi, I., Poiret, T., Zhao, Y., Gultekin, O., He, R., ... & Mattsson, J. (2021). Mesothelin-Specific CAR T Cells Target Ovarian CancerEvaluation of MSLN CAR T Cells for Ovarian Cancer Treatment. Cancer research, 81(11), 3022-3035.

Complete CAR data Funcs

Fig.3 MSLN-directed CAR T-cell infusion augmented survival in an orthotopic mouse model of ovarian cancer.

CAR Construction : m912 scfv-CD28-CD3ζ; m912 scfv-41BB-CD3ζ Latest CAR Construction

Fig.3 MSLN-directed CAR T-cell infusion augmented survival in an orthotopic mouse model of ovarian cancer.

Serial SKOV3 tumor BLI obtained by IVIS of 15 representative mice pre- and posttreatment with CD19z (n = 5), M28z (n = 5) and MBBz (n = 5) CAR T cells. Red crossed images were excluded from the final analysis due to a technical error during BLI measurements.

Schoutrop, E., El-Serafi, I., Poiret, T., Zhao, Y., Gultekin, O., He, R., ... & Mattsson, J. (2021). Mesothelin-Specific CAR T Cells Target Ovarian CancerEvaluation of MSLN CAR T Cells for Ovarian Cancer Treatment. Cancer research, 81(11), 3022-3035.

Complete CAR data Funcs

Fig.4 MSLN-directed CAR T-cell infusion augmented survival in an orthotopic mouse model of ovarian cancer.

CAR Construction : m912 scfv-CD28-CD3ζ; m912 scfv-41BB-CD3ζ Latest CAR Construction

Fig.4 MSLN-directed CAR T-cell infusion augmented survival in an orthotopic mouse model of ovarian cancer.

Kaplan-Meier survival curve.

Schoutrop, E., El-Serafi, I., Poiret, T., Zhao, Y., Gultekin, O., He, R., ... & Mattsson, J. (2021). Mesothelin-Specific CAR T Cells Target Ovarian CancerEvaluation of MSLN CAR T Cells for Ovarian Cancer Treatment. Cancer research, 81(11), 3022-3035.

Complete CAR data FCM

Fig.5 MSLN CAR T cells in mice organs and ex vivo MSLN CAR T-cell functionality.

CAR Construction : m912 scfv-CD28-CD3ζ; m912 scfv-41BB-CD3ζ Latest CAR Construction

Fig.5 MSLN CAR T cells in mice organs and ex vivo MSLN CAR T-cell functionality.

PD-1, LAG3, and TIM3 expression in CAR T cells between different organs in M28z and MBBz mice.

Schoutrop, E., El-Serafi, I., Poiret, T., Zhao, Y., Gultekin, O., He, R., ... & Mattsson, J. (2021). Mesothelin-Specific CAR T Cells Target Ovarian CancerEvaluation of MSLN CAR T Cells for Ovarian Cancer Treatment. Cancer research, 81(11), 3022-3035.

Complete CAR data FCM

Fig.6 MSLN CAR T cells in mice organs and ex vivo MSLN CAR T-cell functionality.

CAR Construction : m912 scfv-CD28-CD3ζ; m912 scfv-41BB-CD3ζ Latest CAR Construction

Fig.6 MSLN CAR T cells in mice organs and ex vivo MSLN CAR T-cell functionality.

PD-1, LAG3, and TIM3 expression by M28z and MBBz CAR T cells rested for one day after collection from the primary ovarian tumor.

Schoutrop, E., El-Serafi, I., Poiret, T., Zhao, Y., Gultekin, O., He, R., ... & Mattsson, J. (2021). Mesothelin-Specific CAR T Cells Target Ovarian CancerEvaluation of MSLN CAR T Cells for Ovarian Cancer Treatment. Cancer research, 81(11), 3022-3035.

Complete CAR data Funcs

Fig.7 MSLN CAR T cells in mice organs and ex vivo MSLN CAR T-cell functionality.

CAR Construction : m912 scfv-CD28-CD3ζ; m912 scfv-41BB-CD3ζ Latest CAR Construction

Fig.7 MSLN CAR T cells in mice organs and ex vivo MSLN CAR T-cell functionality.

Lysis of MSLN+ SKOV3 by M28z and MBBz CAR T cells isolated from tumor, lungs, and spleen after sacrifice and ex vivo resting, measured by a luciferase-based assay.

Schoutrop, E., El-Serafi, I., Poiret, T., Zhao, Y., Gultekin, O., He, R., ... & Mattsson, J. (2021). Mesothelin-Specific CAR T Cells Target Ovarian CancerEvaluation of MSLN CAR T Cells for Ovarian Cancer Treatment. Cancer research, 81(11), 3022-3035.

Complete CAR data Funcs

Fig.8 MSLN CAR T cells in mice organs and ex vivo MSLN CAR T-cell functionality.

CAR Construction : m912 scfv-CD28-CD3ζ; m912 scfv-41BB-CD3ζ Latest CAR Construction

Fig.8 MSLN CAR T cells in mice organs and ex vivo MSLN CAR T-cell functionality.

Levels of IFNγ and TNF in supernatant following ex vivo assays of M28z (n = 8) and MBBz (n = 4) CAR T cells cocultured with MSLN+ SKOV3 cells.

Schoutrop, E., El-Serafi, I., Poiret, T., Zhao, Y., Gultekin, O., He, R., ... & Mattsson, J. (2021). Mesothelin-Specific CAR T Cells Target Ovarian CancerEvaluation of MSLN CAR T Cells for Ovarian Cancer Treatment. Cancer research, 81(11), 3022-3035.

Complete CAR data Funcs

Fig.9 In vitro antitumor efficacy of MSLN CAR transduced T cells

CAR Construction : m912 scfv-CD3ζ; m912 scfv-CD28-CD3ζ Latest CAR Construction

Fig.9 In vitro antitumor efficacy of MSLN CAR transduced T cells

Antigen specific effector function of MSLN CAR-transduced T cells as shown by lysis of MSLN-expressing, target cells measured by chromium-release assays. M912-CAR see M28z (Blue Line)

Adusumilli, P. S., Cherkassky, L., Villena-Vargas, J., Colovos, C., Servais, E., Plotkin, J., ... & Sadelain, M. (2014). Regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science translational medicine, 6(261), 261ra151-261ra151.

Complete CAR data Funcs

Fig.10 Tumor BLI of NOD/SCID/null mice bearing pleural tumor with MSLN CAR transduced T cells.

CAR Construction : m912 scfv-CD3ζ; m912 scfv-CD28-CD3ζ Latest CAR Construction

Fig.10 Tumor BLI of NOD/SCID/null mice bearing pleural tumor with MSLN CAR transduced T cells.

Tumor-bearing mice were treated with either 1E5 (1x) or 3E6 (30x) M28z T cells intravenously (E/T 1:3000 or 1:100, respectively), compared with 1E5 (1x) or 3E5 (3x) M28z T cells intrapleurally (E/T 1:3000 or 1:1000, respectively). (D and F) (M912-CAR see M28z (Blue Line))

Adusumilli, P. S., Cherkassky, L., Villena-Vargas, J., Colovos, C., Servais, E., Plotkin, J., ... & Sadelain, M. (2014). Regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science translational medicine, 6(261), 261ra151-261ra151.

Complete CAR data Funcs

Fig.11 Kaplan-Meier survival analysis of NOD/SCID/null mice bearing pleural tumor with MSLN CAR transduced T cells.

CAR Construction : m912 scfv-CD3ζ; m912 scfv-CD28-CD3ζ Latest CAR Construction

Fig.11 Kaplan-Meier survival analysis of NOD/SCID/null mice bearing pleural tumor with MSLN CAR transduced T cells.

Survival curves were analyzed with log-rank test. (M912-CAR see M28z (Blue Line))

Adusumilli, P. S., Cherkassky, L., Villena-Vargas, J., Colovos, C., Servais, E., Plotkin, J., ... & Sadelain, M. (2014). Regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science translational medicine, 6(261), 261ra151-261ra151.

Complete CAR data Funcs

Fig.12 Intrapleurally administered M28z+ T cells display efficient systemic trafficking and accumulation in extrapleural tumor in an antigen-specific
manner.

CAR Construction : m912 scfv-CD3ζ; m912 scfv-CD28-CD3ζ Latest CAR Construction

Fig.12 Intrapleurally administered M28z+ T cells display efficient systemic trafficking and accumulation in extrapleural tumor in an antigen-specific manner.

Mice with established ffLuc+ MSLN+ tumor in the right flank and pleural cavity and MSLN- tumor in the left flank received Gaussia luciferase+ M28z T cells intrapleurally.

Adusumilli, P. S., Cherkassky, L., Villena-Vargas, J., Colovos, C., Servais, E., Plotkin, J., ... & Sadelain, M. (2014). Regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science translational medicine, 6(261), 261ra151-261ra151.

Complete CAR data Funcs

Fig.13 Intrapleurally administered M28z+ T cells display efficient systemic trafficking and accumulation in extrapleural tumor in an antigen-specific
manner.

CAR Construction : m912 scfv-CD3ζ; m912 scfv-CD28-CD3ζ Latest CAR Construction

Fig.13 Intrapleurally administered M28z+ T cells display efficient systemic trafficking and accumulation in extrapleural tumor in an antigen-specific manner.

Intrapleurally administered M28z+ T cells show early and robust accumulation in MSLN+ intraperitoneal tumor, compared with intravenously administered T cells.

Adusumilli, P. S., Cherkassky, L., Villena-Vargas, J., Colovos, C., Servais, E., Plotkin, J., ... & Sadelain, M. (2014). Regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science translational medicine, 6(261), 261ra151-261ra151.

Complete CAR data Funcs

Fig.14 Intrapleurally administered M28z+ T cells display efficient systemic trafficking and accumulation in extrapleural tumor in an antigen-specific manner.

CAR Construction : m912 scfv-CD3ζ; m912 scfv-CD28-CD3ζ Latest CAR Construction

Fig.14 Intrapleurally administered M28z+ T cells display efficient systemic trafficking and accumulation in extrapleural tumor in an antigen-specific manner.

Quantification of the fold increase in signal intensity of the peritoneal cavity in tumor-bearing mice displays enhanced T cell accumulation with intrapleural administration, compared with intravenous administration (n = 3 per group)

Adusumilli, P. S., Cherkassky, L., Villena-Vargas, J., Colovos, C., Servais, E., Plotkin, J., ... & Sadelain, M. (2014). Regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science translational medicine, 6(261), 261ra151-261ra151.

Complete CAR data Funcs

Fig.15 Intrapleurally administered M28z T cells eradicate pleural tumor and establish long-term CD4+ predominant persistence.

CAR Construction : m912 scfv-CD3ζ; m912 scfv-CD28-CD3ζ Latest CAR Construction

Fig.15 Intrapleurally administered M28z T cells eradicate pleural tumor and establish long-term CD4+ predominant persistence.

s. In total, 1E5CAR+ Mz, M28z, or P28z (negative control) T cells were intrapleurally administered into mice bearing established tumors. Left: Tumor burden. Right: Kaplan-Meier survival curve.

Adusumilli, P. S., Cherkassky, L., Villena-Vargas, J., Colovos, C., Servais, E., Plotkin, J., ... & Sadelain, M. (2014). Regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science translational medicine, 6(261), 261ra151-261ra151.

Complete CAR data Funcs

Fig.16 Intrapleurally administered M28z T cells eradicate pleural tumor and establish long-term CD4+ predominant persistence.

CAR Construction : m912 scfv-CD3ζ; m912 scfv-CD28-CD3ζ Latest CAR Construction

Fig.16 Intrapleurally administered M28z T cells eradicate pleural tumor and establish long-term CD4+ predominant persistence.

CD28 costimulation enhances CAR+ T cell persistence. Absolute CAR+ T cell counts (per milliliter of peripheral blood) at 50 days after intrapleural administration of 3 × 106 CAR+ T cells.

Adusumilli, P. S., Cherkassky, L., Villena-Vargas, J., Colovos, C., Servais, E., Plotkin, J., ... & Sadelain, M. (2014). Regional delivery of mesothelin-targeted CAR T cell therapy generates potent and long-lasting CD4-dependent tumor immunity. Science translational medicine, 6(261), 261ra151-261ra151.

More Published Data More Published Data

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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