A transgene-encoded cell surface polypeptide for selection, in vivo tracking, and ablation of engineered cells

• Published in: Blood Vol. 118; no. 5; pp. 1255 - 1263
• Main Authors: Wang, Xiuli, Chang, Wen-Chung, Wong, ChingLam W., Colcher, David, Sherman, Mark, Ostberg, Julie R., Forman, Stephen J., Riddell, Stanley R., Jensen, Michael C.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 04.08.2011; Americain Society of Hematology; American Society of Hematology
• Subjects: Animals; Antigens, Surface - genetics; Apoptosis - genetics; Biological and medical sciences; Cell Separation - methods; Cell Tracking - methods; Cells - metabolism; Cells - pathology; Cells, Cultured; Gene Therapy; Genes, erbB-1; Genes, Reporter; Genes, Transgenic, Suicide - physiology; Hematologic and hematopoietic diseases; Humans; Interleukin Receptor Common gamma Subunit - genetics; Medical sciences; Mice; Mice, Inbred NOD; Mice, SCID; Mice, Transgenic; Peptides - genetics; Tissue Engineering - methods; Transgenes - genetics; In vivo; Hematology; Polypeptide; Cell surface; Transgene; Selection
• ISSN: 0006-4971; 1528-0020; 1528-0020
• DOI: 10.1182/blood-2011-02-337360

An unmet need in cell engineering is the availability of a single transgene encoded, functionally inert, human polypeptide that can serve multiple purposes, including ex vivo cell selection, in vivo cell tracking, and as a target for in vivo cell ablation. Here we describe a truncated human EGFR polypeptide (huEGFRt) that is devoid of extracellular N-terminal ligand binding domains and intracellular receptor tyrosine kinase activity but retains the native amino acid sequence, type I transmembrane cell surface localization, and a conformationally intact binding epitope for pharmaceutical-grade anti-EGFR monoclonal antibody, cetuximab (Erbitux). After lentiviral transduction of human T cells with vectors that coordinately express tumor-specific chimeric antigen receptors and huEGFRt, we show that huEGFRt serves as a highly efficient selection epitope for chimeric antigen receptor+ T cells using biotinylated cetuximab in conjunction with current good manufacturing practices (cGMP)-grade anti-biotin immunomagnetic microbeads. Moreover, huEGFRt provides a cell surface marker for in vivo tracking of adoptively transferred T cells using both flow cytometry and immunohistochemistry, and a target for cetuximab-mediated antibody-dependent cellular cytotoxicity and in vivo elimination. The versatility of huEGFRt and the availability of pharmaceutical-grade reagents for its clinical application denote huEGFRt as a significant new tool for cellular engineering.