Pytlak 2023 MiP2023

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MiPschool 2023

Pytlak 2023 MiP2023

Mitochondrial defect in human bronchial epithelial cells lacking the BKCa channel.

Link: MiP2023 Obergurgl AT

Pytlak Karolina (2023)

Event: MiP2023 Obergurgl AT

Authors: Pytlak Karolina, Maliszewska – Olejniczak K, Sek Aleksandra, Szewczyk Adam, Bednarczyk Piotr, Kulawiak Bogusz

Human bronchial epithelial (HBE) cells form an external barrier in the airways and are constantly exposed to factors such as urban dust.
Recently, the large conductance calcium-activated potassium (mitoBKCa) channel has been identified in the inner mitochondrial membrane of HBE cells. The pore-forming subunit of the channel is encoded by the KCNMA1 gene, which also encodes plasma membrane BKCa channels. Mitochondrial potassium channels regulate mitochondrial membrane potential, oxygen consumption, mitochondrial volume and reactive oxygen species synthesis. Activation of mitoBKCa induces cytoprotection of cardiac and brain tissue.
In our project, we applied CRISPR/Cas9 technique to disrupt KCNMA1 gene in the HBE cell line (16HBE14o- cells). The newly formed line showed no mitoBKCa channel activity. We also noticed changes related to the deregulation of the cell cycle. The loss of mitoBKCa significantly affected mitochondrial function. We observed a decrease in the rate of mitochondrial respiration. Furthermore, we analyzed the organization of respiratory chain complexes using Blue Native electrophoresis. In addition, analysis of the expression of selected genes encoding mitochondrial proteins showed changes in cells with disrupted KCNMA1 gene. Nevertheless, a thorough understanding of the observed mitochondrial dysfunction requires further study. We conclude that the presence of the mitoBKCa channel in HBE cells is essential for the preservation of mitochondrial function and is important for the proper function of these cells as part of the human airways.

Affiliations and acknowledgements

Pytlak K1, Maliszewska – Olejniczak K2, SΔ™k A1, Szewczyk A1, Bednarczyk P2, Kulawiak B1
  1. Lab Intracellular Ion Channels, Nencki Inst Experimental Biol, Warsaw, Poland
  2. Dept Biophysics, Warsaw Univ Life Sciences, Poland

Funding: This study was supported by a grant 2019/35/B/NZ1/02546 from the National Science Centre in Poland.


Tissue;cell: Lung;gill 

Event: Poster 

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