Small protein blockers of human IL-6 receptor alpha inhibit proliferation and migration of cancer cells

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F24%3A00586070" target="_blank" >RIV/61388971:_____/24:00586070 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/86652036:_____/24:00586070 RIV/61989592:15110/24:73627272 RIV/44555601:13440/24:43898632 RIV/00216208:11110/24:10482400 RIV/00098892:_____/24:10158624

Výsledek na webu

<a href="https://biosignaling.biomedcentral.com/articles/10.1186/s12964-024-01630-w" target="_blank" >https://biosignaling.biomedcentral.com/articles/10.1186/s12964-024-01630-w</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1186/s12964-024-01630-w" target="_blank" >10.1186/s12964-024-01630-w</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Small protein blockers of human IL-6 receptor alpha inhibit proliferation and migration of cancer cells

Popis výsledku v původním jazyce

Background Interleukin-6 (IL-6) is a multifunctional cytokine that controls the immune response, and its role has been described in the development of autoimmune diseases. Signaling via its cognate IL-6 receptor (IL-6R) complex is critical in tumor progression and, therefore, IL-6R represents an important therapeutic target.Methods An albumin-binding domain-derived highly complex combinatorial library was used to select IL-6R alpha (IL-6R alpha)-targeted small protein binders using ribosome display. Large-scale screening of bacterial lysates of individual clones was performed using ELISA, and their IL-6R alpha blocking potential was verified by competition ELISA. The binding of proteins to cells was monitored by flow cytometry and confocal microscopy on HEK293T-transfected cells, and inhibition of signaling function was examined using HEK-Blue IL-6 reporter cells. Protein binding kinetics to living cells was measured by LigandTracer, cell proliferation and toxicity by iCELLigence and Incucyte, cell migration by the scratch wound healing assay, and prediction of binding poses using molecular modeling by docking.Results We demonstrated a collection of protein variants called NEF ligands, selected from an albumin-binding domain scaffold-derived combinatorial library, and showed their binding specificity to human IL-6R alpha and antagonistic effect in HEK-Blue IL-6 reporter cells. The three most promising NEF108, NEF163, and NEF172 variants inhibited cell proliferation of malignant melanoma (G361 and A2058) and pancreatic (PaTu and MiaPaCa) cancer cells, and suppressed migration of malignant melanoma (A2058), pancreatic carcinoma (PaTu), and glioblastoma (GAMG) cells in vitro. The NEF binders also recognized maturation-induced IL-6R alpha expression and interfered with IL-6-induced differentiation in primary human B cells.Conclusion We report on the generation of small protein blockers of human IL-6R alpha using directed evolution. NEF proteins represent a promising class of non-toxic anti-tumor agents with migrastatic potential.

Název v anglickém jazyce

Small protein blockers of human IL-6 receptor alpha inhibit proliferation and migration of cancer cells

Popis výsledku anglicky

Background Interleukin-6 (IL-6) is a multifunctional cytokine that controls the immune response, and its role has been described in the development of autoimmune diseases. Signaling via its cognate IL-6 receptor (IL-6R) complex is critical in tumor progression and, therefore, IL-6R represents an important therapeutic target.Methods An albumin-binding domain-derived highly complex combinatorial library was used to select IL-6R alpha (IL-6R alpha)-targeted small protein binders using ribosome display. Large-scale screening of bacterial lysates of individual clones was performed using ELISA, and their IL-6R alpha blocking potential was verified by competition ELISA. The binding of proteins to cells was monitored by flow cytometry and confocal microscopy on HEK293T-transfected cells, and inhibition of signaling function was examined using HEK-Blue IL-6 reporter cells. Protein binding kinetics to living cells was measured by LigandTracer, cell proliferation and toxicity by iCELLigence and Incucyte, cell migration by the scratch wound healing assay, and prediction of binding poses using molecular modeling by docking.Results We demonstrated a collection of protein variants called NEF ligands, selected from an albumin-binding domain scaffold-derived combinatorial library, and showed their binding specificity to human IL-6R alpha and antagonistic effect in HEK-Blue IL-6 reporter cells. The three most promising NEF108, NEF163, and NEF172 variants inhibited cell proliferation of malignant melanoma (G361 and A2058) and pancreatic (PaTu and MiaPaCa) cancer cells, and suppressed migration of malignant melanoma (A2058), pancreatic carcinoma (PaTu), and glioblastoma (GAMG) cells in vitro. The NEF binders also recognized maturation-induced IL-6R alpha expression and interfered with IL-6-induced differentiation in primary human B cells.Conclusion We report on the generation of small protein blockers of human IL-6R alpha using directed evolution. NEF proteins represent a promising class of non-toxic anti-tumor agents with migrastatic potential.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10601 - Cell biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Cell communication and signaling : CCS

ISSN

1478-811X

e-ISSN

1478-811X

Svazek periodika

22

Číslo periodika v rámci svazku

1

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

26

Strana od-do

261

Kód UT WoS článku

001215466500002

EID výsledku v databázi Scopus

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