Manfred Schwab
`Gri Keye
`
`Miltenyi Ex. 1010 Page1
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`Encyclopedia of
`Serle
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`VOLUME 1
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`AE
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`Y) Saeeran
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`combined with cytokine, such as immunocytokine, to
`deliver co-stimulatory signal
`through endogenous
`cytokine receptors. One of the major advantages of the
`strategy infusing CAR* T cells lies in the modular
`composition of the CAR molecule that combines an
`antigen-binding domain with signaling domains for
`effector-cell activation. This allows investigators to
`swap the CAR exodomain with a scFv or ligand that
`recognizes or binds to a desired cell-surface antigen.
`Future experiments will build on the catalog of antigen-
`targeting receptors to evaluate expression level, density,
`and stability of the CAR expression on the T-cell
`surface, as well as affinity of the binding domain for
`antigen in the context of tumor targets with varying
`antigen density. These studies will impact CAR-mediated
`immunotherapy since
`low-density antigen-positive
`tumorcells or low affinity CAR maylead to emergence
`oftumorescape.Just as genetic engineering can be used
`to alter the CAR exodomain, so the transmembrane, or
`endodomain may be altered to provide a fully-
`competent antigen-dependent T-cell activation signal.
`The majority of the CARs generated harbor CD3-C€
`signaling chain which is currently considered more
`efficient in activating T cells for cytolysis, compared to
`chimeric FceRI-y. Other CARs have been generated
`activating T cells through syk, Ick, but the full-effect of
`these receptors with respect to cellular activation and
`stability ofreceptor expression onthe cell surface has yet
`to be determined. Similarly the CAR endodomain has
`been altered to express a co-stimulatory signaling
`molecule,
`such as chimeric CD28,
`to provide a
`coordinated signal with CD3-¢ to provide a more
`complete T-cell activation signal
`than achieved by
`signaling through CD3-{ alone. Because different types
`of co-stimulation may result in different patterns of
`cellular activation,it will likely be beneficial to explore
`alternative co-stimulatory pathways in CAR-mediated
`T-cell activation. Increasingly, investigators are devel-
`oping the tools to genetically modify T cells using
`techniques that cause minimal manipulation of the
`productleaving intact the full range ofT-cell homing and
`proliferation potentials. Finally, genetic modification is
`being used to accomplish more than redirect T-cell
`specificity. For example, experiments are underway to
`render the T cells resistant to the anti-inflammatory and
`deleterious effects ofiatrogenic glucocorticoids and TGF-
`B secreted by tumorcell.
`
`Conclusion
`Althoughlimitations remain, genetic modifications enable
`investigators to engineer T cells with augmented
`
`References
`
`1. Eshhar Z, Waks T, Bendavid A et al. (2001) Functional
`expression of chimeric receptor genes in humanT cells. J
`Immunol Methods 248(1—2):67—76
`2. Rossig C, Brenner MK (2003) Chimeric T-cell receptors
`for the targeting of cancer cells. Acta Haematol 110
`(203):154-159
`3. Cooper L, Topp MS, Serrano LM et al. (2003) T-cell
`clones can be rendered specific for CD19:
`selective augmentation of the graft-versus-B-lineage
`leukemia effect. Blood 101:1637-1644
`4. Morgan RA, Dudley ME, Wunderlich JR et al. (2006)
`Cancer regression in patients after transfer of genetically
`engineered lymphocytes. Science 314:126-—129
`5. Park JR, Digiusto DL, Slovak M etal. (2007) Adoptive
`transfer of chimeric antigen receptor re-directed cytolytic
`T lymphocyte clones in patients with neuroblastoma. Mol
`Ther 15(4):825-833
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`Chimeric Genes
`
`>» Fusion Genes
`
`Chimeric Oncogenes
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`» Fusion Genes
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`Chimeric Oncoproteins
`
`Definition
`Chimeric proteins generated by » chromosometranslo-
`cations of two cellular genes in which the functional
`domainsoftwo separate genes are fused together and/or
`alter regulation of gene expression. Many chimeric
`oncoproteins characterized to date appear to act as
`aberrant
`transcription factors,
`likely functioning in
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