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Anti-EphA2 (1F12)-FcRɣ+PI3K CAR-MA (CARMA-W0654)

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

The vector of anti-EphA2 chimeric antigen receptor (CAR) is constructed for the engineering of macrophage cells to target human EphA2 and induce target cell phagocytosis. The macrophage cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-EphA2 antibody (1F12) linked to FcRɣ+PI3K signaling domains. The vector product was designed for the treatment of GBM, glioma.

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Details

  • Target
  • EphA2
  • Cell Type
  • Macrophage
  • Targeting Diseases
  • GBM, glioma
  • Vector Name
  • pCDCAR1
  • Vector length
  • ~8kb
  • Vector Type
  • Lentiviral vector
  • Size
  • 10 µg
  • Storage
  • Store at -20°C for long term.

CAR Components

  • Promoter
  • EF1a
  • scFv
  • 1F12
  • Hinge
  • CD8
  • TM
  • CD8
  • ICD
  • FcRɣ+PI3K
  • Marker
  • GFP or N/A
  • Receptor Construction
  • scFv-hinge(CD8)-TM(CD8)-ICD(FcRɣ+PI3K)
  • Discription of Signaling Cassetes
  • FcRɣ
    FcRɣ is used as the CAR intracellular signaling domain. As a phagocytosis receptor, FcRɣ has cytosolic Immunoreceptor Tyrosine-based Activation Motifs (ITAMs) that are phosphorylated by Src family kinases. Based on this mechanism, FcRɣ signaling can induce macrophage phagocytosis which has been demonstrated in the published article.
    Fc gamma receptors belong to the group of non-catalytic tyrosine-phosphorylated receptors which share a similar signalling pathway involving phosphorylation of tyrosine residues. The receptors generate signals within their cells through an important activation motif known as an immunoreceptor tyrosine-based activation motif (ITAM). An ITAM is present in the intracellular tail of FcγRIIA, and its phosphorylation induces phagocytosis in macrophages. FcγRI and FcγRIIIA do not have an ITAM but can transmit an activating signal to their phagocytes by interacting with another protein that does. This adaptor protein is called the Fcγ subunit and, like FcγRIIA, contains the two YXXL sequences that are characteristic of an ITAM.

    PI3K
    Previous work demonstrated that PI3K signaling is important for internalization of large targets. To increase PI3K recruitment to the CAR-P, we fused the portion of the CD19 cytoplasmic domain (amino acids 500 to 534) that recruits the p85 subunit of PI3K to the CAR. A CAR-P containing the p85 recruitment motif alone (CAR-P-PI3K) was able to induce some whole cell engulfment. Assembling an array of motifs designed to recruit distinct phagocytic effectors can increase CAR-P activity.

Target

  • Target
  • EphA2
  • Target Species
  • Human
  • Clone
  • 1F12
  • Host
  • Human
  • Gene Name
  • EPH receptor A2
  • Introduction
  • This gene belongs to the ephrin receptor subfamily of the protein-tyrosine kinase family. EPH and EPH-related receptors have been implicated in mediating developmental events, particularly in the nervous system. Receptors in the EPH subfamily typically have a single kinase domain and an extracellular region containing a Cys-rich domain and 2 fibronectin type III repeats. The ephrin receptors are divided into 2 groups based on the similarity of their extracellular domain sequences and their affinities for binding ephrin-A and ephrin-B ligands. This gene encodes a protein that binds ephrin-A ligands. Mutations in this gene are the cause of certain genetically-related cataract disorders.
  • Alternative Names
  • ECK; CTPA; ARCC2; CTPP1; CTRCT6; EPHA2; EPH receptor A2; Erythropoetin producing hepatocellular carcinoma A2

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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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