Engineering and Therapeutic Application of Single-Chain Bivalent TGF-β Family Traps

Zwaagstra, John C.; Sulea, Traian; Baardsnes, Jason; Lenferink, Anne E.G.; Collins, Cathy; Cantin, Christiane; Paul-Roc, Béatrice; Grothé, Suzanne; Hossain, Sorif; Richer, Louis-Philippe; L’Abbé, Denis; Tom, Roseanne; Cass, Brian; Durocher, Yves; O’Connor-McCourt, Maureen D.

doi:10.1158/1535-7163.mct-12-0060

Cited by 17 publications

(21 citation statements)

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“…We previously found that TGF-β plays a role in negatively regulating NK cell function by antagonizing positive signaling pathways (Yu et al, 2006) and, subsequently, other groups found that TGF-β also acts as an extrinsic negative regulator in NK cell development (Marcoe et al, 2012). Of note, inhibition of TGF-β signaling with neutralizing antibodies and other approaches has been used to treat cancer (Bouquet et al, 2011; Liu et al, 2012; Zwaagstra et al, 2012). …”

Section: Discussionmentioning

confidence: 99%

Transcription Factor Foxo1 Is a Negative Regulator of Natural Killer Cell Maturation and Function

et al. 2015

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SUMMARY Little is known about the role of negative regulators in controlling natural killer (NK) cell development and effector functions. Foxo1 is a multifunctional transcription factor of the forkhead family. Using a mouse model of conditional deletion in NK cells, we found that Foxo1 negatively controlled NK cell differentiation and function. Immature NK cells expressed abundant Foxo1 and little Tbx21 relative to mature NK cells, but these two transcription factors reversed their expression as NK cells proceeded through development. Foxo1 promoted NK cell homing to lymph nodes through upregulating CD62L expression, and impaired late-stage maturation and effector functions by repressing Tbx21 expression. Loss of Foxo1 rescued the defect in late-stage NK cell maturation in heterozygous Tbx21+/− mice. Collectively, our data reveal a regulatory pathway by which the negative regulator Foxo1 and the positive regulator Tbx21 play opposing roles in controlling NK cell development and effector functions.

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Section: Discussionmentioning

confidence: 99%

Transcription Factor Foxo1 Is a Negative Regulator of Natural Killer Cell Maturation and Function

et al. 2015

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“…3B). While the tandem TβRII-ECD TGF-β trap and sTβRII have been shown to neutralize TGF-β1 to similar degrees (21), sTβRII molecules consist of two TβRII-ECD domains linked by mouse IgG 2a in a chimeric fashion. In our in vitro and in vivo systems where Fc-receptor expressing cells are present, sTβRII may be internalized and rendered ineffective, compared to FIST15, which contains no such domains.…”

Section: Discussionmentioning

confidence: 99%

“…We found that the addition of the IL15Rα- sushi domain to IL15 in the FIST15 fusion protein enhances its biological activity and rescues its signaling activity compared to a fusion of IL15 to the TGF-β ligand trap without the IL15Rα- sushi domain (FIST15Δsushi). The use of two tandem TGF-β receptor ectodomains (type II; TβRII-ECD) recently described (21) and modified for use in FIST15, allows it to effectively neutralize TGF-β signaling. FIST15 treatment enhanced the capacity of NK and CD8 + T cells to produce critical anti-tumor cytokines, such as IFNγ and TNFα, as well as their expression of cytolytic effector molecules.…”

Section: Introductionmentioning

confidence: 99%

Stimulation of Natural Killer Cell–Mediated Tumor Immunity by an IL15/TGFβ–Neutralizing Fusion Protein

Deng

Chinnadurai

et al. 2016

Cancer Research

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The clinical efficacy of immune cytokines used for cancer therapy is hampered by elements of the immunosuppressive tumor microenvironment such as TGF-ß. Here we demonstrate that FIST15, a recombinant chimeric protein composed of the T cell stimulatory cytokine IL-15, the sushi domain of IL15Rα and a TGF-β ligand trap, can overcome immunosuppressive TGF-β to effectively stimulate the proliferation and activation of natural killer (NK) and CD8+ T cells with potent antitumor properties. FIST15-treated NK and CD8+ T cells produced more IFNγ and TNFα compared to treatment with IL-15 and a commercially available TGF-β receptor-Fc fusion protein (sTβRII) in the presence of TGF-β. Murine B16 melanoma cells which overproduce TGF-β were lysed by FIST15-treated NK cells in vitro at doses ~10-fold lower than NK cells treated with IL-15 and sTβRII. Melanoma cells transduced to express FIST15 failed to establish tumors in vivo in immunocompetent murine hosts and could only form tumors in beige mice lacking NK cells. Mice injected with the same cells were also protected from subsequent challenge by unmodified B16 melanoma cells. Lastly, mice with pre-established B16 melanoma tumors responded to FIST15 treatment more strongly compared to tumors treated with control cytokines. Taken together, our results offer a preclinical proof of concept for the use of FIST15 as a new class of biological therapeutics that can coordinately neutralize the effects of immunosuppressive TGF-β in the tumor microvironment while empowering tumor immunity.

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“…S1) and (ii) a subset of the Abs elicited in response to immunization competed with soluble dimeric type II TGF-β receptor ectodomain [(TβRII) 2 ; Zwaagstra et al, 2012] for TGF-β3 binding, since binding of (TβRII) 2 to TGF-β3 was diminished in the presence of serum from the TGF-β3-immunized llama ( Supplementary Fig. S2).…”

Section: Resultsmentioning

confidence: 99%

“…Specifically, we constructed a phage-displayed sdAb library from the lymphocytes of a llama immunized with TGF-β3 and panned the library in a single round against TGF-β3. We eluted sdAbs competitively with either soluble TGF-β receptor ectodomain (Zwaagstra et al,2012 ) or an irrelevant competitor, used NGS of panning outputs to identify sdAbs specifically displaced by the former biomolecule, and characterized the resulting sdAbs using binding and in vitro cytokine neutralization assays (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

Isolation of TGF-β-neutralizing single-domain antibodies of predetermined epitope specificity using next-generation DNA sequencing

Henry

Hussack

Collins

et al. 2016

Protein Engineering, Design and Selection

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Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n'arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. Questions? Contact the NRC Publications Archive team atPublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. If you wish to email the authors directly, please see the first page of the publication for their contact information. NRC Publications Archive Archives des publications du CNRCThis publication could be one of several versions: author's original, accepted manuscript or the publisher's version. / La version de cette publication peut être l'une des suivantes : la version prépublication de l'auteur, la version acceptée du manuscrit ou la version de l'éditeur. NRC Publications Record / Notice d'Archives des publications de CNRC:http://nparc.cisti-icist.nrc-cnrc.gc.ca/eng/view/object/?id=646f2540-63b6-4b0e-a7ae-731f5c2a6082 http://nparc.cisti-icist.nrc-cnrc.gc.ca/fra/voir/objet/?id=646f2540-63b6-4b0e-a7ae-731f5c2a6082 Protein Engineering, Design & Selection, 2016, pp. 1-5 doi: 10.1093/protein/gzw043 AbstractThe epitope specificity of therapeutic antibodies is often critical to their efficacy and mode of action. Here, we report the isolation of single-domain antibodies (sdAbs) against a pre-specified epitope of TGF-β3: namely, the site of interaction between the cytokine and its cell-surface type II receptor. By panning a phage-displayed immune llama VHH library against TGF-β3 using competitive elution with soluble dimeric type II receptor ectodomain in tandem with next-generation DNA sequencing, we identified several sdAbs that competed with the receptor for TGF-β3 binding and neutralized TGF-β3i nin vitro cellular assays. In contrast, all other sdAbs identified using conventional panning approaches (i.e., without regard to epitope specificity) did not target the site of receptor:cytokine interaction. We expect this strategy to be generally applicable for identifying epitope-specific sdAbs when binding reagents directed against the epitope of interest are available. The sdAbs identified here are of potential interest as cancer immunotherapeutics.

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Engineering and Therapeutic Application of Single-Chain Bivalent TGF-β Family Traps

Cited by 17 publications

References 50 publications

Transcription Factor Foxo1 Is a Negative Regulator of Natural Killer Cell Maturation and Function

Transcription Factor Foxo1 Is a Negative Regulator of Natural Killer Cell Maturation and Function

Stimulation of Natural Killer Cell–Mediated Tumor Immunity by an IL15/TGFβ–Neutralizing Fusion Protein

Isolation of TGF-β-neutralizing single-domain antibodies of predetermined epitope specificity using next-generation DNA sequencing

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