Molecular dissection of the interactions of an antitumor interleukin‐2‐derived mutein on a phage display‐based platform

Rojas, Gertrudis; Carmenate, Tania; León, Kalet

doi:10.1002/jmr.2440

Cited by 7 publications

(3 citation statements)

References 41 publications

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“…Although the initial reports of phage-displayed biologically active IL-2 were published more than 20 years ago 27,28 , this platform has only recently been exploited to map the interactions of hundreds of IL-2-derived variants 29–31 , and IL-2 engineering has been dominated by yeast display 11,32 . Here we report directed molecular evolution of phage-displayed IL-2 resulting in the discovery of single mutations that increase display levels, enhance secretion by human host cells and diminish IL-2 aggregation.…”

Section: Introductionmentioning

confidence: 99%

Directed evolution of super-secreted variants from phage-displayed human Interleukin-2

Rojas

Carmenate

Santo-Tomás

et al. 2019

Sci Rep

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Selection from a phage display library derived from human Interleukin-2 (IL-2) yielded mutated variants with greatly enhanced display levels of the functional cytokine on filamentous phages. Introduction of a single amino acid replacement selected that way (K35E) increased the secretion levels of IL-2-containing fusion proteins from human transfected host cells up to 20-fold. Super-secreted (K35E) IL-2/Fc is biologically active in vitro and in vivo, has anti-tumor activity and exhibits a remarkable reduction in its aggregation propensity- the major manufacturability issue limiting IL-2 usefulness up to now. Improvement of secretion was also shown for a panel of IL-2-engineered variants with altered receptor binding properties, including a selective agonist and a super agonist that kept their unique properties. Our findings will improve developability of the growing family of IL-2-derived immunotherapeutic agents and could have a broader impact on the engineering of structurally related four-alpha-helix bundle cytokines.

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Section: Introductionmentioning

confidence: 99%

Directed evolution of super-secreted variants from phage-displayed human Interleukin-2

Rojas

Carmenate

Santo-Tomás

et al. 2019

Sci Rep

Self Cite

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“…By introducing a particular set of mutations along the binding interface of IL-2 with the alpha subunit it is possible to disrupt their interaction. This new IL-2 mutein can, therefore, cause a decrease in CD25+ cells while still expanding CD8+ and Natural Killer cells 12 . A highly efficient strategy for creating this new muteins is through the recently awarded Novel Prize technique, Phage Display.…”

Section: Il2 R α Regulationmentioning

confidence: 99%

Therapeutics and prospects of Interleukin 2

Lovato

Padilla

2019

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Interleukin-2 was discovered back in 1983 as an autocrine growth factor for cultured T cells and was the first biological product created through the use of recombinant DNA. IL-2 tumor immunotherapy performed the first historical clinical demonstration of the possibility to cause an effective anticancer immune reaction, mediated by cytotoxic lymphocytes activated from IL-2 stimulation. The Interleukin 2 receptor is a heterotrimeric protein that is composed of three peptide chains: the alpha chain, the beta chain and the gamma chain of the common cytokine receptor. There are 3 majors’ ways of interfering with the IL-2/IL-2R to use it as treatments: Antibodies, Aptamers, and punctual mutagenesis. Recent studies have shown, that Il-2 therapies for cancer, specifically targets restoring the individual’s natural antitumor immune response. HIV directed treatments have demonstrated the necessity of introducing the IL-2 complemented with the patient’s antiretroviral therapy.

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“…31 Engagement of this dimeric receptor by IL-2 also triggers activation and proliferation signals. [32][33][34] A human IL-2 mutant whose interaction with CD25 (IL-2Rα chain) is significantly disrupted, 35 that behaves as an IL-2R signaling agonist which expands preferentially CD8 + T lymphocytes and NK cells over Tregs, has been developed at the Center of Molecular Immunology (CIM). 28,36 This IL-2 variant, termed no-alpha mutein, has demonstrated a higher anti-metastatic effect than IL-2 in 3LL-D122 and B16 tumor models.…”

Section: Introductionmentioning

confidence: 99%

A rationally-engineered IL-2 improves the antitumor effect of anti-CD20 therapy

et al. 2020

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Anti-CD20 treatment represents a therapeutic benefit for patients with B-cell lymphomas, although more efficient therapies are needed for refractory or relapsing patients. Among them, the combination of anti-CD20 and IL-2 that induces T cell response has been hampered by the expansion of FoxP3 + Tregs that strongly express the high affinity IL-2 receptor (IL-2R αβγ). We explore here the anti-tumor effect of an anti-CD20 antibody combined with a mutated IL-2 (no-alpha mutein) which has a disrupted affinity for the IL-2R αβγ. We demonstrate that anti-CD20/no-alpha mutein combination significantly augments the survival rate of mice challenged with huCD20 + cells as compared to animals treated with anti-CD20 ± IL-2. Moreover, the combination with no-alpha mutein but not IL-2 provokes an increase of granzyme B and perforin in splenic NK and CD8 + T cells, a reduction of Tregs and an increase in activated macrophages. The former combination also induces a T helper profile different from that obtained with IL-2, with an earlier polarization to Th1 and no increase in Th17. The therapeutic effect of anti-CD20/noalpha mutein was accompanied by an expansion of peripheral central (TCM) and effector (TEM) memory CD8 + T cell compartments. Last, as opposed to IL-2, no-alpha mutein administered at the beginning of anti-CD20 treatment did not dampen the long-term protection of surviving mice after tumor rechallenge. Thus, this study shows that the combination of anti-tumor antibodies and no-alpha mutein is a promising approach to improve the therapeutic effect of these antibodies by potentiating NK/macrophagemediated innate immunity and the adaptive T-cell response.

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Molecular dissection of the interactions of an antitumor interleukin‐2‐derived mutein on a phage display‐based platform

Cited by 7 publications

References 41 publications

Directed evolution of super-secreted variants from phage-displayed human Interleukin-2

Directed evolution of super-secreted variants from phage-displayed human Interleukin-2

Therapeutics and prospects of Interleukin 2

A rationally-engineered IL-2 improves the antitumor effect of anti-CD20 therapy

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