Udział kanału VDAC w patogenezie choroby Huntingtona w oparciu o badania modeli komórkowych
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2017
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VDAC contribution to pathogenesis of Huntington disease revealed by studies of the disease cell models
Abstract
Celem niniejszej pracy było sprawdzenie czy białko VDAC jest istotne dla rozwoju choroby Huntingtona i zbadanie znaczenia obecności izoform białka VDAC dla obserwowanych efektów ekspresji Htt i mHtt. W badaniach stosowano dwa modele HD: linie komórkowe PC12 i model oparty na komórkach S. cerevisiae. Oba te modele umożliwiają kontrolowaną ekspresję Htt i mHtt, przy czym drugi z nich umożliwia także badanie wpływu tej ekspresji w obecności bądź przy braku danej izoformy ludzkiego białka VDAC. Modele te wykorzystano dla zbadania wpływu ekspresji Htt i mHtt na przeżywalność komórek, funkcje bioenergetyczne mitochondriów, a także właściwości kanałowe białek VDAC.
Otrzymane wyniki wskazują, że obecność białka VDAC ma znaczenie dla analizowanych efektów ekspresji Htt i mHtt. Co więcej, znaczenie ma także izoforma badanego białka VDAC. Stwierdzono na przykład, że ekspresja mHtt w komórkach PC12, w których dominuje ilościowo izoforma VDAC1, jak i w komórkach modelu drożdżowego z ekspresją VDAC1 człowieka skutkuje zmniejszeniem przeżywalności tych komórek, przy czym w przypadku komórek drożdży efekt ten wymaga obecności stresu temperaturowego (tj. 37°C). Co ciekawe, ów stres temperaturowy może mieć wpływ na poziom agregacji mHtt. Otrzymane wyniki uzyskane dla komórek modelu drożdżowego z ekspresją hVDAC1 wydają się potwierdzać kluczowe znaczenie tej izoformy VDAC dla efektu mHtt na funkcjonowanie mitochondriów. Pozostaje to w związku z wynikami uzyskanymi dla preparatów VDAC izolowanych z komórek PC12, w przypadku których wykazano, że ekspresja mHtt może prowadzić do zmian aktywności kanałowej białka VDAC1.
The aim of this study was to check whether VDAC protein is important for Htt and/or mHtt effects on cells, which in turn would indicate VDAC participation in the pathogenesis of HD. The contribution of VDAC isoforms to the observed effects of Htt and mHtt expression was also evaluated. Two models of this disease were used in the study: PC12 and Saccharomyces cerevisiae cells. Both of these models allow for controlled expression of Htt and mHtt, while the second one also allows for determination of the effect of Htt/mHtt expression in the presence or absence of human VDAC isoforms. These models were used to investigate the effect of Htt and mHtt expression on cell survival, bioenergetic functions as well as channel properties of VDAC proteins. The obtained results indicate that the presence of VDAC protein is important for the analyzed effects of Htt and mHtt expression. Moreover, the isoform of VDAC protein also matters. For example, the expression of mHtt in PC12 cells (in which VDAC1 isoform is quantitatively dominant), as well as in the yeast HD model cells expressing human VDAC1 (hVDAC1) results in decreased survival of these cells. In the case of the yeast cells this effect requires the presence of temperature stress (37oC), which may affect the level of mHtt aggregation. Moreover, data obtained for hVDAC1 expressing cells of the yeast HD model appear to confirm the key importance of this VDAC isoform for mHtt effect on mitochondrial function. This is in line with results obtained for VDAC preparations isolated from PC12 cells as it was observed that mHtt expression may lead to changes in VDAC1 channel activity.
The aim of this study was to check whether VDAC protein is important for Htt and/or mHtt effects on cells, which in turn would indicate VDAC participation in the pathogenesis of HD. The contribution of VDAC isoforms to the observed effects of Htt and mHtt expression was also evaluated. Two models of this disease were used in the study: PC12 and Saccharomyces cerevisiae cells. Both of these models allow for controlled expression of Htt and mHtt, while the second one also allows for determination of the effect of Htt/mHtt expression in the presence or absence of human VDAC isoforms. These models were used to investigate the effect of Htt and mHtt expression on cell survival, bioenergetic functions as well as channel properties of VDAC proteins. The obtained results indicate that the presence of VDAC protein is important for the analyzed effects of Htt and mHtt expression. Moreover, the isoform of VDAC protein also matters. For example, the expression of mHtt in PC12 cells (in which VDAC1 isoform is quantitatively dominant), as well as in the yeast HD model cells expressing human VDAC1 (hVDAC1) results in decreased survival of these cells. In the case of the yeast cells this effect requires the presence of temperature stress (37oC), which may affect the level of mHtt aggregation. Moreover, data obtained for hVDAC1 expressing cells of the yeast HD model appear to confirm the key importance of this VDAC isoform for mHtt effect on mitochondrial function. This is in line with results obtained for VDAC preparations isolated from PC12 cells as it was observed that mHtt expression may lead to changes in VDAC1 channel activity.
Description
Wydział Biologii
Sponsor
Badania zrealizowano ze środków:
NCN 2011/01/B/NZ3/00359
GDWB-06/2015
KNOW RNA Research Centre in Poznan No.01/KNOW2/2014
Keywords
choroba Huntingtona, Huntington disease, mitochondria, VDAC