ER-stress induce epithelial-to-mesenchymal transition in
expandable lung epithelia derived from pluripotent stem cells

a 2023

ER-stress induce epithelial-to-mesenchymal transition in expandable lung epithelia derived from pluripotent stem cells

PORTAKAL, Türkan, Jarmila HERŮDKOVÁ, Vendula PELKOVÁ, Lukáš MORÁŇ, Veronika SEDLÁKOVÁ et. al.

Základní údaje

Originální název

ER-stress induce epithelial-to-mesenchymal transition in expandable lung epithelia derived from pluripotent stem cells

Autoři

PORTAKAL, Türkan (792 Turecko, domácí), Jarmila HERŮDKOVÁ (703 Slovensko, domácí), Vendula PELKOVÁ (203 Česká republika, domácí), Lukáš MORÁŇ (203 Česká republika, domácí), Veronika SEDLÁKOVÁ (203 Česká republika, domácí), Volodymyr POROKH (804 Ukrajina, domácí), Vítězslav HAVLÍČEK (203 Česká republika, domácí), Hana KOTASOVÁ (203 Česká republika, domácí), Aleš HAMPL (203 Česká republika, domácí) a Petr VAŇHARA (203 Česká republika, garant, domácí)

Vydání

BIOCEV Regeneration III, 2023

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

30402 Technologies involving the manipulation of cells, tissues, organs or the whole organism

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Kód RIV

RIV/00216224:14110/23:00134300

Organizační jednotka

Lékařská fakulta

Klíčová slova anglicky

expandable lung epithelia; lung cell differentiation; ER stress; EMT

Příznaky

Mezinárodní význam

Změněno: 24. 11. 2023 07:53, doc. RNDr. Petr Vaňhara, Ph.D.

Anotace

V originále

Introduction (max. 800 characters): Lung cells are constantly exposed to stress signals, which can contribute to various diseases. The endoplasmic reticulum (ER) plays a crucial role in protein synthesis and cell stress response, when accumulation of misfolded proteins triggers a general stress signaling pathway - unfolded protein stress response (UPR). However, the exact impact of UPR on the growth, differentiation, and regeneration of lung cells is not fully understood. Expandable lung epithelial (ELEP) cells are derived from human embryonic stem cells and provide a unique model for studying lung biology and regeneration. The proposed hypothesis suggests that ER stress induces significant changes in ELEP cells, such as epithelial-mesenchymal transition (EMT), through increased expression of UPR proteins. Materials and methods (max. 1000 characters):* ELEP cells were differentiated from human embryonic stem cell (hESC) lines as described previously. Briefly, hESCs were differentiated into distal foregut endoderm through the activin signaling. Then lung-specific differentiation was then mediated by FGF7, FGF10, BMPs, and WNT signaling pathways. Retinoic acid and dexamethasone were used for the maturation and acquisition of functional characteristics of the ELEP cells. Acquisition of differentiated phenotype of ELEP cells was confirmed by TTF1 expression by flow cytometry and western blotting. Viability was estimated by determination of the conversion rate of the tetrazolium dye (MTT) to the insoluble formazan. mRNA and protein levels were determined by qRT-PCR and western blotting, respectively. Subcellular localization of major regulators was determined by immunofluorescent microscopy. Results and conclusions (max. 700 characters):* ELEP cells showed high sensitivity to accumulation of misfolded proteins in the ER by N-clycosylation inhibition, and high rate of cell death. Induction of ER stress was confirmed by upregulation of major ER chaperon BiP and downstream transcription factor CHOP. In addition, we observed increase of E-cadherin signal in cytoplasm, suggesting accumulation of misfolded protein the ER,while N-cadherin retained the membrane signal, even upon ER stress. Induction of ER stress also induced changes in cell morphology, from the cobblestone, epithelial towards the elongated, mesenchymal phenotype. In summary, we showed that ER stress in ELEP cells induced EMT through the activation of the UPR pathway. Acknowledgements (max. 200 characters):* This research is supported by the Czech Science Foundation, project no. GA23-06675S, and Masaryk University, project no. MUNI/A/1301/2022.

Návaznosti

GA23-06675S, projekt VaV

Název: Plicní stres a regenerace

Investor: Grantová agentura ČR, Plicní stres a regenerace

MUNI/A/1301/2022, interní kód MU

Název: Zdroje pro tkáňové inženýrství 13

Investor: Masarykova univerzita, Zdroje pro tkáňové inženýrství 13

Citovat

PORTAKAL, Türkan, Jarmila HERŮDKOVÁ, Vendula PELKOVÁ, Lukáš MORÁŇ, Veronika SEDLÁKOVÁ, Volodymyr POROKH, Vítězslav HAVLÍČEK, Hana KOTASOVÁ, Aleš HAMPL a Petr VAŇHARA. ER-stress induce epithelial-to-mesenchymal transition in expandable lung epithelia derived from pluripotent stem cells. In BIOCEV Regeneration III. 2023.

@proceedings{2342319,
   author = {Portakal, Türkan and Herůdková, Jarmila and Pelková, Vendula and Moráň, Lukáš and Sedláková, Veronika and Porokh, Volodymyr and Havlíček, Vítězslav and Kotasová, Hana and Hampl, Aleš and Vaňhara, Petr},
   booktitle = {BIOCEV Regeneration III},
   keywords = {expandable lung epithelia; lung cell differentiation; ER stress; EMT},
   language = {eng},
   title = {ER-stress induce epithelial-to-mesenchymal transition in expandable lung epithelia derived from pluripotent stem cells},
   year = {2023}
}

TY  - CONF
ID  - 2342319
AU  - Portakal, Türkan - Herůdková, Jarmila - Pelková, Vendula - Moráň, Lukáš - Sedláková, Veronika - Porokh, Volodymyr - Havlíček, Vítězslav - Kotasová, Hana - Hampl, Aleš - Vaňhara, Petr
PY  - 2023
TI  - ER-stress induce epithelial-to-mesenchymal transition in expandable lung epithelia derived from pluripotent stem cells
KW  - expandable lung epithelia
KW  - lung cell differentiation
KW  - ER stress
KW  - EMT
N2  - Introduction (max. 800 characters): Lung cells are constantly exposed to stress signals, which can contribute to various diseases. The endoplasmic reticulum (ER) plays a crucial role in protein synthesis and cell stress response, when accumulation of misfolded proteins triggers a general stress signaling pathway - unfolded protein stress response (UPR). However, the exact impact of UPR on the growth, differentiation, and regeneration of lung cells is not fully understood. Expandable lung epithelial (ELEP) cells are derived from human embryonic stem cells and provide a unique model for studying lung biology and regeneration. The proposed hypothesis suggests that ER stress induces significant changes in ELEP cells, such as epithelial-mesenchymal transition (EMT), through increased expression of UPR proteins. Materials and methods (max. 1000 characters):* ELEP cells were differentiated from human embryonic stem cell (hESC) lines as described previously. Briefly, hESCs were differentiated into distal foregut endoderm through the activin signaling. Then lung-specific differentiation was then mediated by FGF7, FGF10, BMPs, and WNT signaling pathways. Retinoic acid and dexamethasone were used for the maturation and acquisition of functional characteristics of the ELEP cells. Acquisition of differentiated phenotype of ELEP cells was confirmed by TTF1 expression by flow cytometry and western blotting. Viability was estimated by determination of the conversion rate of the tetrazolium dye (MTT) to the insoluble formazan. mRNA and protein levels were determined by qRT-PCR and western blotting, respectively. Subcellular localization of major regulators was determined by immunofluorescent microscopy. Results and conclusions (max. 700 characters):* ELEP cells showed high sensitivity to accumulation of misfolded proteins in the ER by N-clycosylation inhibition, and high rate of cell death. Induction of ER stress was confirmed by upregulation of major ER chaperon BiP and downstream transcription factor CHOP. In addition, we observed increase of E-cadherin signal in cytoplasm, suggesting accumulation of misfolded protein the ER,while N-cadherin retained the membrane signal, even upon ER stress. Induction of ER stress also induced changes in cell morphology, from the cobblestone, epithelial towards the elongated, mesenchymal phenotype. In summary, we showed that ER stress in ELEP cells induced EMT through the activation of the UPR pathway. Acknowledgements (max. 200 characters):* This research is supported by the Czech Science Foundation, project no. GA23-06675S, and Masaryk University, project no. MUNI/A/1301/2022.
ER  -

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