Cytosolic delivery of monobodies using the bacterial type III secretion system inhibits oncogenic BCR: ABL1 signaling

Cell Communication and Signaling, Journal Year: 2024, Volume and Issue: 22(1)

Published: Oct. 16, 2024

The inability of biologics to pass the plasma membrane prevents their development as therapeutics for intracellular targets. To address lack methods cytosolic protein delivery, we used type III secretion system (T3SS) Y. enterocolitica, which naturally injects bacterial proteins into eukaryotic host cells, deliver monobody cancer cells. Monobodies are small synthetic binding that can inhibit oncogene signaling in cells with high selectivity upon expression. Here, engineered monobodies targeting BCR::ABL1 tyrosine kinase efficient delivery by T3SS, quantified and target engagement monitored inhibition signaling.

Language: Английский

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Multifunctional sorafenib-loaded MXene for enhanced cancer therapy: In vitro and in vivo study based on chemotherapy/photothermal therapy approach

International Journal of Pharmaceutics, Journal Year: 2025, Volume and Issue: unknown, P. 125492 - 125492

Published: March 1, 2025

Language: Английский

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Development of mirror-image monobodies targeting the oncogenic BCR::ABL1 kinase

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Dec. 23, 2024

Abstract Mirror-image proteins, composed of d -amino acids, are an attractive therapeutic modality, as they exhibit high metabolic stability and lack immunogenicity. Development mirror-image binding proteins is achieved through chemical synthesis -target phage display library selection l -binders (mirror-image) that consequently bind the physiological -targets. Monobodies well-established synthetic ( -)binding their small size (~90 residues) endogenous cysteine residues make them particularly accessible to synthesis. Here, we develop monobodies with nanomolar affinities against -SH2 domain leukemic tyrosine kinase BCR::ABL1. Two crystal structures heterochiral monobody-SH2 complexes reveal targeting pY pocket by unconventional mode. We then prepare potent -monobodies either ligating two chemically synthesized -peptides or self-assembly without ligation. Their proper folding determined high-affinity shown. protease-resistant, show long-term plasma stability, inhibit BCR::ABL1 activity in cell lysates permeabilized cells. Hence, demonstrate functional can be developed readily. Our work represents important step towards possible future use when combined emerging methods enable cytoplasmic delivery monobodies.

Language: Английский

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Development of mirror-image monobodies targeting the oncogenic BCR::ABL1 kinase

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: April 26, 2024

Mirror-image proteins, which are composed of d-amino acids, an attractive therapeutic modality, as they exhibit high metabolic stability and lack immunogenicity. Development mirror-image binding proteins is achieved through chemical synthesis the d-target protein, phage display library selection l-binder (mirror-image) d-binder that consequently binds physiological l-target. Monobodies among most well-established synthetic (l-)binding their small size (~90 residues) endogenous cysteine residues make them particularly accessible to synthesis. Here we developed monobodies with nanomolar affinities against d-SH2 domain leukemic tyrosine kinase BCR::ABL1. Two crystal structures heterochiral monobody-SH2 complexes revealed targeting pY pocket by unconventional mode. We then prepared stable potent d-monobodies either ligating two chemically synthesized d-peptides or self-assembly without ligation. Their proper folding were determined affinity l-target was shown. protease-resistant, showed long-term plasma stability, inhibited BCR::ABL1 activity bound in cells. Hence, demonstrate functional can be readily, enabling use future d-protein therapeutics target a broad spectrum protein-protein interactions.

Language: Английский

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Cytosolic delivery of monobodies using the bacterial type III secretion system inhibits oncogenic BCR::ABL1 signaling

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 5, 2024

Background The inability of biologics to pass the plasma membrane prevents their development as therapeutics for intracellular targets. To address lack methods cytosolic protein delivery, we used type III secretion system (T3SS) Y. enterocolitica, which naturally injects bacterial proteins into eukaryotic host cells, deliver monobody cancer cells. Monobodies are small synthetic binding that can inhibit oncogene signaling in cells with high selectivity upon expression. Here, engineered monobodies targeting BCR::ABL1 tyrosine kinase efficient delivery by T3SS, quantified and target engagement monitored inhibition signaling. Methods In vitro assays were performed characterize destabilized (thermal shift assay isothermal titration calorimetry) assess T3SS. Immunoblot study translocation different cell lines determine concentration after translocation. Split-Nanoluc understand degradation kinetics evaluate Phospho flow cytometry apoptosis functional effects BCR:ABL1-expressing leukemia Results enable stable mutant retained affinity efficiently injected lines. After injection, concentrations reached mid-micromolar considerably exceeding affinity. We found selectively engaged cytosol. resulted oncogenic specifically induced BCR::ABL1-dependent consistent phenotype when same was intracellularly expressed. Conclusion Hence, establish T3SS enterocolitica a highly method enabling selective pathways providing foundation future therapeutic application against

Language: Английский

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Non-immunoglobin scaffold binders as efficient affinity ligands for purification of broad-spectrum serum albumins

Talanta, Journal Year: 2024, Volume and Issue: 285, P. 127262 - 127262

Published: Nov. 22, 2024

Language: Английский

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