Primary Human T Lymphocytes Engineered with a Codon-Optimized IL-15 Gene Resist Cytokine Withdrawal-Induced Apoptosis and Persist Long-Term in the Absence of Exogenous Cytokine

Hsu, Cary; Hughes, Marybeth S.; Zheng, Zhili; Bray, Regina; Rosenberg, Steven A.; Morgan, Richard A.

doi:10.4049/jimmunol.175.11.7226

Cited by 85 publications

(71 citation statements)

References 52 publications

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“…8 This retroviral vector contains the murine stem cell virus LTR and RNA processing signals similar to the MFG-class of retroviral vectors. The anti-MART-1 AIB TCR retroviral vector was used for control transductions.…”

Section: Generation Of the Lc15 Cell Line Vector Transduction And Cmentioning

confidence: 99%

Cytokine-independent growth and clonal expansion of a primary human CD8+ T-cell clone following retroviral transduction with the IL-15 gene

Hsu

Jones

Cohen

et al. 2007

Blood

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100

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Malignancies arising from retrovirally transduced hematopoietic stem cells have been reported in animal models and human gene therapy trials. Whether mature lymphocytes are susceptible to insertional mutagenesis is unknown. We have characterized a primary human CD8 ؉ Tcell clone, which exhibited logarithmic ex vivo growth in the absence of exogenous cytokine support for more than 1 year after transduction with a murine leukemia virus-based vector encoding the T-cell growth factor IL-15. Phenotypically, the clone was CD28 ؊ , CD45RA ؊ , CD45RO ؉ , and CD62L ؊ , a profile consistent with effector memory T lymphocytes. After gene transfer with tumor-antigenspecific T-cell receptors, the clone secreted IFN-␥ upon encountering tumor targets, providing further evidence that they derived from mature lymphocytes. IntroductionHuman gene therapy strategies using replication-incompetent retroviral vectors were implemented cautiously because of many concerns, including the potential for adverse events resulting from vector insertions in the human genome. 1 Theoretical concerns about insertional mutagenesis became reality with the report that 3 of 11 patients with severe combined immunodeficiency-X1 (SCID-X1) developed T-cell leukemia after treatment with hematopoietic stem cells (HSCs) engineered with the common ␥-chain cytokine receptor gene. These leukemias resulted from integration events which activated the LMO2 proto-oncogene. 2 Nevertheless, after more than 17 years of clinical gene therapy trials this remains the only report of overt cancer development resulting from genemodified cell therapy. Our understanding of the pathogenesis of human malignancy resulting from insertional mutagenesis remains limited to this specific clinical scenario.Several observations from the SCID-X1 gene therapy trial fall in line with general principles established through animal models and other human clinical trials investigating retroviral gene transfer. The well-characterized murine leukemia virus (MLV)-based gene transfer vectors integrate preferentially in the vicinity of cellular promoters, 3,4 potentially increasing the probability of activating or inactivating vector insertions which may dysregulate gene expression. 5,6 The target cells most at risk of insertional mutagenesis are believed to be primitive progenitor cells. 7 Potentially, therapeutic transgenes may have leukemogenic properties in the appropriate setting. 6 The interactions between retroviral gene transfer vectors and hematopoietic cells are being investigated intensely.In this report, we have characterized a primary human T-cell clone, which exhibited continuous growth after transduction with an MLV-based retroviral vector encoding the T-cell growth factor, IL-15. This was an unexpected and intriguing finding for several reasons. First, human cells are inherently difficult to immortalize, and the process of transformation in human cells is believed to be considerably more complex than that of experimental animals. Second, the cells that were transduced consisted of...

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Section: Generation Of the Lc15 Cell Line Vector Transduction And Cmentioning

confidence: 99%

Cytokine-independent growth and clonal expansion of a primary human CD8+ T-cell clone following retroviral transduction with the IL-15 gene

Hsu

Jones

Cohen

et al. 2007

Blood

Self Cite

100

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“…Although IL-2 plays a critical role in the stimulation of activated T cells, it is also involved in activationinduced cell death and the establishment of peripheral tolerance. Several studies have indicated that an alternative γ-chain cytokine, IL-15, provides potent homeostatic T-cell survival/proliferative signals, inhibits IL-2-mediated activation-induced cell death, and may be superior to IL-2 in immunotherapy applications (25,26). Recently, IL-15 has been shown to function in establishing the long-term persistence of adoptively transferred central memory T (T CM ) cells in primates, suggesting significant potential in T-cell therapy for cancer (27).…”

Section: A Clinically Relevant T-cell Proliferation System With Drug-mentioning

confidence: 99%

Genetic control of mammalian T-cell proliferation with synthetic RNA regulatory systems

Chen

Jensen

Smolke

2010

Proc. Natl. Acad. Sci. U.S.A.

242

245

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RNA molecules perform diverse regulatory functions in natural biological systems, and numerous synthetic RNA-based control devices that integrate sensing and gene-regulatory functions have been demonstrated, predominantly in bacteria and yeast. Despite potential advantages of RNA-based genetic control strategies in clinical applications, there has been limited success in extending engineered RNA devices to mammalian gene-expression control and no example of their application to functional response regulation in mammalian systems. Here we describe a synthetic RNA-based regulatory system and its application in advancing cellular therapies by linking rationally designed, drug-responsive, ribozymebased regulatory devices to growth cytokine targets to control mouse and primary human T-cell proliferation. We further demonstrate the ability of our synthetic controllers to effectively modulate T-cell growth rate in response to drug input in vivo. Our RNAbased regulatory system exhibits unique properties critical for translation to therapeutic applications, including adaptability to diverse ligand inputs and regulatory targets, tunable regulatory stringency, and rapid response to input availability. By providing tight gene-expression control with customizable ligand inputs, RNA-based regulatory systems can greatly improve cellular therapies and advance broad applications in health and medicine. T he ability to control functional responses in mammalian cells with customizable and compact regulatory systems in vivo addresses a critical need in diverse clinical applications, particularly in cellular therapies (1). As an example, adoptive T-cell therapy seeks to harness the precision and efficacy of the immune system against diseases that escape the body's natural surveillance. The adoptive transfer of antigen-specific T cells can reconstitute immunity to viruses and mediate tumor regression (2-4). T cells engineered to express tumor-specific T-cell antigen receptors can achieve highly refined target recognition (5), thus minimizing toxic off-target effects associated with conventional chemotherapy. However, considerable research has shown that the persistence of transferred T cells in vivo is both central to therapeutic success and elusive to current technology (6, 7). The efficacy of adoptive immunotherapy in humans is often limited by the failure of transferred T cells to survive in the host (8, 9).Clonal expansion of T cells is a critical component of T-cell activation mediated by cytokines such as IL-2 and IL-15, which activate JAK-STAT signaling pathways and lead to the expression of genes involved in growth modulation (10) (Fig. 1A). Sustaining the survival and proliferation of Tcells following adoptive transfer is challenging because of the limited availability of homeostatic cytokines (IL-15/IL-7) and stimulatory antigen presenting cells. State-of-the-art strategies for improving the persistence of adoptively transferred lymphocytes require that patients be subjected to myeloablative total body irradiation/chemother...

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“…We can increase homing to tumor sites by expressing chemokine receptors specific for the molecules the tumors produce, while T cell expansion and persistence in the hostile tumor microenvironment can be increased by expressing transgenes that confer resistance to apoptosis, senescence, or inhibitory cytokines, or that provide autocrine T cell growth. 38 Any genetic strategy that enhances the survival of T cells, however, runs the risk of producing unwanted lymphoproliferation. Although protocols transferring genes to T cells have been safe to date, this strategy is being implemented with caution.…”

Section: Modification Of Effector T Cellsmentioning

confidence: 99%

Progress and Prospects: Gene Therapy Clinical Trials (Part 2)

Griesenbach

2007

Gene Ther

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Primary Human T Lymphocytes Engineered with a Codon-Optimized IL-15 Gene Resist Cytokine Withdrawal-Induced Apoptosis and Persist Long-Term in the Absence of Exogenous Cytokine

Cited by 85 publications

References 52 publications

Cytokine-independent growth and clonal expansion of a primary human CD8+ T-cell clone following retroviral transduction with the IL-15 gene

Cytokine-independent growth and clonal expansion of a primary human CD8+ T-cell clone following retroviral transduction with the IL-15 gene

Genetic control of mammalian T-cell proliferation with synthetic RNA regulatory systems

Progress and Prospects: Gene Therapy Clinical Trials (Part 2)

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