Light-enhanced cytotoxicity and intracellular trafficking of the PD-L1-targeting photoimmunoconjugate EITC-atezolizumab

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11160%2F25%3A10513891" target="_blank" >RIV/00216208:11160/25:10513891 - isvavai.cz</a>

Výsledek na webu

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=8bznW3fKcK" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=8bznW3fKcK</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.biopha.2025.118550" target="_blank" >10.1016/j.biopha.2025.118550</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Light-enhanced cytotoxicity and intracellular trafficking of the PD-L1-targeting photoimmunoconjugate EITC-atezolizumab

Popis výsledku v původním jazyce

Background: Photodynamic therapy (PDT) uses light to generate reactive oxygen species (ROS) that initiate cytotoxic responses in cancer cells. Photoimmunotherapy (PIT) improves PDT specificity by linking photosensitizers (PS) to monoclonal antibodies. A promising monoclonal antibody (mAb) for immunoconjugates is atezolizumab (Tecentriq), an EMA/FDA-approved PD-L1 immune checkpoint inhibitor for treating non-small cell lung cancer (NSCLC). Objective: Our goal was to: Optimize the cytotoxic efficacy of a photoimmunoconjugate of eosin-5-isothiocyanate and atezolizumab (EITC-atezolizumab) in NSCLC cells; Study the uptake and intracellular transport of atezolizumab; Evaluate EITC-atezolizumab as a candidate for photochemical internalization (PCI) of the ribosome-inactivating protein gelonin. Methods: Cytotoxic efficacy of EITC-atezolizumab was evaluated in NSCLC cell lines H1975 (PD-L1-high) and A549 (PD-L1-low) using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Confocal microscopy was employed to observe the uptake and intracellular localization of AlexaFluor647-atezolizumab and its colocalization with either LysoTracker(TM) Blue or an anti-CD63 antibody labelled with Alexa488. Results: EITC-atezolizumab PIT significantly enhanced cytotoxicity, showing much higher sensitivity in H1975 than A549 NSCLC cells. PIT with EITC-atezolizumab did not enhance gelonin efficacy. Confocal microscopy of H1975 cells revealed near-plasmamembrane puncta fluorescence of Alexa647-atezolizumab, with fractions clustering in CD63+ organelles peaking at 6 h. In A549 cells, AlexaFluor647-atezolizumab showed sustained accumulation in LysoTracker(TM) Blue+ vesicles for up to 24 h. Conclusion: To the best of our knowledge, this is the first documentation demonstrating that atezolizumab is transported to CD63-positive organelles, thereby enhancing our understanding of its intracellular trafficking. Our study also strengthens the concept of PIT and atezolizumab-based targeting of PD-L1+ NSCLCs.

Název v anglickém jazyce

Light-enhanced cytotoxicity and intracellular trafficking of the PD-L1-targeting photoimmunoconjugate EITC-atezolizumab

Popis výsledku anglicky

Background: Photodynamic therapy (PDT) uses light to generate reactive oxygen species (ROS) that initiate cytotoxic responses in cancer cells. Photoimmunotherapy (PIT) improves PDT specificity by linking photosensitizers (PS) to monoclonal antibodies. A promising monoclonal antibody (mAb) for immunoconjugates is atezolizumab (Tecentriq), an EMA/FDA-approved PD-L1 immune checkpoint inhibitor for treating non-small cell lung cancer (NSCLC). Objective: Our goal was to: Optimize the cytotoxic efficacy of a photoimmunoconjugate of eosin-5-isothiocyanate and atezolizumab (EITC-atezolizumab) in NSCLC cells; Study the uptake and intracellular transport of atezolizumab; Evaluate EITC-atezolizumab as a candidate for photochemical internalization (PCI) of the ribosome-inactivating protein gelonin. Methods: Cytotoxic efficacy of EITC-atezolizumab was evaluated in NSCLC cell lines H1975 (PD-L1-high) and A549 (PD-L1-low) using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Confocal microscopy was employed to observe the uptake and intracellular localization of AlexaFluor647-atezolizumab and its colocalization with either LysoTracker(TM) Blue or an anti-CD63 antibody labelled with Alexa488. Results: EITC-atezolizumab PIT significantly enhanced cytotoxicity, showing much higher sensitivity in H1975 than A549 NSCLC cells. PIT with EITC-atezolizumab did not enhance gelonin efficacy. Confocal microscopy of H1975 cells revealed near-plasmamembrane puncta fluorescence of Alexa647-atezolizumab, with fractions clustering in CD63+ organelles peaking at 6 h. In A549 cells, AlexaFluor647-atezolizumab showed sustained accumulation in LysoTracker(TM) Blue+ vesicles for up to 24 h. Conclusion: To the best of our knowledge, this is the first documentation demonstrating that atezolizumab is transported to CD63-positive organelles, thereby enhancing our understanding of its intracellular trafficking. Our study also strengthens the concept of PIT and atezolizumab-based targeting of PD-L1+ NSCLCs.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30104 - Pharmacology and pharmacy

Projekt

—

Návaznosti

S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

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

Název periodika

Biomedicine &amp; Pharmacotherapy

ISSN

0753-3322

e-ISSN

1950-6007

Svazek periodika

191

Číslo periodika v rámci svazku

October

Stát vydavatele periodika

FR - Francouzská republika

Počet stran výsledku

11

Strana od-do

118550

Kód UT WoS článku

999

EID výsledku v databázi Scopus

2-s2.0-105015598905