title: Chimeric receptor mRNA transfection as a tool to generate antineoplastic lymphocytes.
abstract: mRNA transfection is a useful approach for temporal cell reprogramming with minimal risk of transgene-mediated mutagenesis. We applied this to redirect lymphocyte cytotoxicity toward malignant cells. Using the chimeric immune receptor (CIR) constructs anti-CD19 CIR and 8H9 CIR, we achieved uniform expression of CIRs on virtually the entire population of lymphocytes. We reprogrammed CD3+ CD8+, CD3+ CD4+, and natural killer (NK ) cells toward autologous and allogeneic targets such as B cells, Daudi lymphoma, primary melanoma, breast ductal carcinoma, breast adenocarcinoma, and rhabdomyosarcoma. The reprogramming procedure is fast. Although most of the experiments were performed on lymphocytes obtained after 7-day activation, only 1-day activation of T cells with anti-CD3, anti-CD28 antibodies, and interleukin-2 is sufficient to develop both lymphocyte cytotoxicity and competence for mRNA transfer. The entire procedure, which includes lymphocyte activation and reprogramming, can be completed in 2 days. The efficiency of mRNA-modified human T cells was tested in a murine xenograft model. Human CD3+CD8+ lymphocytes expressing anti-CD19 CIR mRNA inhibited Daudi lymphoma growth in NOD=SCID mice. These results demonstrate that a mixed population of cytotoxic lymphocytes, including T cells together with NK cells, can be quickly and simultaneously reprogrammed by mRNA against autologous malignancies. With relatively minor modifications the described method of lymphocyte reprogramming can be scaled up for cancer therapy.
authors.affiliation: Department of Genetics, Yale University School of Medicine, New Haven, CT 06520, USA.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Rabinovich
authors.forename: Marina E
authors.initials: ME
authors.lastname: Komarovskaya
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Wrzesinski
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Alderman
authors.forename: Tulin
authors.initials: T
authors.lastname: Budak-Alpdogan
authors.forename: Alexander
authors.initials: A
authors.lastname: Karpikov
authors.forename: Hongfen
authors.initials: H
authors.lastname: Guo
authors.forename: Richard A
authors.initials: RA
authors.lastname: Flavell
authors.forename: Nai-Kong
authors.initials: NK
authors.lastname: Cheung
authors.forename: Sherman M
authors.initials: SM
authors.lastname: Weissman
authors.forename: Erkut
authors.initials: E
authors.lastname: Bahceci
mesh_terms: D000818:Animals; D018941:Antigens, CD19 / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D005260:Female; D006801:Humans; D008213:Lymphocyte Activation; D008223:Lymphoma / Q000235:genetics / Q000276:immunology*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection; D019076:Transgenes; D023041:Xenograft Model Antitumor Assays
chemical_list: D018941:Antigens, CD19; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins