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Ethanol-mediated activation of the NLRP3 inflammasome in iPS cells and iPS cells-derived neural progenitor cells.

De Filippis L, Halikere A, McGowan H, Moore JC, Tischfield JA, Hart RP, Pang ZP - Mol Brain (2016)

Bottom Line: Ethanol exposure for 24 hours or 7 days does not affect the proliferation of iPS cells and NPCs, but primes an innate immune-like response by activating the NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway.This leads to an increase of microtubule-associated protein 1A/1B-light chain 3(+) (LC3B(+)) autophagic puncta and impairment of the mitochondrial and lysosomal distribution.In addition, a decrease of mature neurons derived from differentiating NPCs is evident in ethanol pre-exposed compared to control NPCs.

View Article: PubMed Central - PubMed

Affiliation: Child Health Institute of New Jersey, Rutgers University-Robert Wood Johnson Medical School, room 3233D, 89 French Street, New Brunswick, NJ, 08901, USA. ld473@rwjms.rutgers.edu.

ABSTRACT

Background: Alcohol abuse produces an enormous impact on health, society, and the economy. Currently, there are very limited therapies available, largely due to the poor understanding of mechanisms underlying alcohol use disorders (AUDs) in humans. Oxidative damage of mitochondria and cellular proteins aggravates the progression of neuroinflammation and neurological disorders initiated by alcohol abuse.

Results: Here we show that ethanol exposure causes neuroinflammation in both human induced pluripotent stem (iPS) cells and human neural progenitor cells (NPCs). Ethanol exposure for 24 hours or 7 days does not affect the proliferation of iPS cells and NPCs, but primes an innate immune-like response by activating the NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway. This leads to an increase of microtubule-associated protein 1A/1B-light chain 3(+) (LC3B(+)) autophagic puncta and impairment of the mitochondrial and lysosomal distribution. In addition, a decrease of mature neurons derived from differentiating NPCs is evident in ethanol pre-exposed compared to control NPCs. Moreover, a second insult of a pro-inflammatory factor in addition to ethanol preexposure enhances innate cellular inflammation in human iPS cells.

Conclusions: This study provides strong evidence that neuronal inflammation contributes to the pathophysiology of AUDs through the activation of the inflammasome pathway in human cellular models.

No MeSH data available.


Related in: MedlinePlus

Ethanol alters mitochondrial patterns in iPS cells, NPCs, and NPC-derived neurons. Confocal microscopy images showing the mitochondrial pattern (Mitotracker) in iPS cells (a), NPCs (b), and NPC-derived neurons (after 26 days of differentiation) (c) after treatment with ethanol for 24hr or 7d. Nuclei (in blue) are counterstained with DAPI. Inserts: co-immunolabeling with MAP2 shows different colocalization of Mitotracker in neuronal cells. Scale bars: 10 μm
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Fig4: Ethanol alters mitochondrial patterns in iPS cells, NPCs, and NPC-derived neurons. Confocal microscopy images showing the mitochondrial pattern (Mitotracker) in iPS cells (a), NPCs (b), and NPC-derived neurons (after 26 days of differentiation) (c) after treatment with ethanol for 24hr or 7d. Nuclei (in blue) are counterstained with DAPI. Inserts: co-immunolabeling with MAP2 shows different colocalization of Mitotracker in neuronal cells. Scale bars: 10 μm

Mentions: Consistent with previous studies showing cross talk between the inflammasome pathway and lysosomal-mitochondrial machinery [8], we investigated the mitochondrial and lysosomal patterning in iPS cells and NPCs with or without ethanol exposure, as well as in neurons derived from NPCs with or without ethanol pre-exposure. In particular, we investigated the distribution of mitochondria and lysosomes using a Mitotracker Assay™ (Fig. 4) and IHC for the lysosomal marker Lamp1 (Fig. 5), respectively. Both mitochondria (Fig. 4) and lysosomes (Fig. 5) appear to be reduced in iPS cells or NPCs exposed to ethanol. In contrast to the more dispersed mitochondrial distribution in untreated iPS cells, NPCs, or neurons, the mitochondria cluster more prominently in the perinuclear area in treated cells, with a particularly robust effect in the 7d condition (Fig. 4a, b and 5a, b). Similar effects were observed in neurons derived from NPCs that were pre-exposed to ethanol (Fig. 4c and 5c).Fig. 4


Ethanol-mediated activation of the NLRP3 inflammasome in iPS cells and iPS cells-derived neural progenitor cells.

De Filippis L, Halikere A, McGowan H, Moore JC, Tischfield JA, Hart RP, Pang ZP - Mol Brain (2016)

Ethanol alters mitochondrial patterns in iPS cells, NPCs, and NPC-derived neurons. Confocal microscopy images showing the mitochondrial pattern (Mitotracker) in iPS cells (a), NPCs (b), and NPC-derived neurons (after 26 days of differentiation) (c) after treatment with ethanol for 24hr or 7d. Nuclei (in blue) are counterstained with DAPI. Inserts: co-immunolabeling with MAP2 shows different colocalization of Mitotracker in neuronal cells. Scale bars: 10 μm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4862119&req=5

Fig4: Ethanol alters mitochondrial patterns in iPS cells, NPCs, and NPC-derived neurons. Confocal microscopy images showing the mitochondrial pattern (Mitotracker) in iPS cells (a), NPCs (b), and NPC-derived neurons (after 26 days of differentiation) (c) after treatment with ethanol for 24hr or 7d. Nuclei (in blue) are counterstained with DAPI. Inserts: co-immunolabeling with MAP2 shows different colocalization of Mitotracker in neuronal cells. Scale bars: 10 μm
Mentions: Consistent with previous studies showing cross talk between the inflammasome pathway and lysosomal-mitochondrial machinery [8], we investigated the mitochondrial and lysosomal patterning in iPS cells and NPCs with or without ethanol exposure, as well as in neurons derived from NPCs with or without ethanol pre-exposure. In particular, we investigated the distribution of mitochondria and lysosomes using a Mitotracker Assay™ (Fig. 4) and IHC for the lysosomal marker Lamp1 (Fig. 5), respectively. Both mitochondria (Fig. 4) and lysosomes (Fig. 5) appear to be reduced in iPS cells or NPCs exposed to ethanol. In contrast to the more dispersed mitochondrial distribution in untreated iPS cells, NPCs, or neurons, the mitochondria cluster more prominently in the perinuclear area in treated cells, with a particularly robust effect in the 7d condition (Fig. 4a, b and 5a, b). Similar effects were observed in neurons derived from NPCs that were pre-exposed to ethanol (Fig. 4c and 5c).Fig. 4

Bottom Line: Ethanol exposure for 24 hours or 7 days does not affect the proliferation of iPS cells and NPCs, but primes an innate immune-like response by activating the NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway.This leads to an increase of microtubule-associated protein 1A/1B-light chain 3(+) (LC3B(+)) autophagic puncta and impairment of the mitochondrial and lysosomal distribution.In addition, a decrease of mature neurons derived from differentiating NPCs is evident in ethanol pre-exposed compared to control NPCs.

View Article: PubMed Central - PubMed

Affiliation: Child Health Institute of New Jersey, Rutgers University-Robert Wood Johnson Medical School, room 3233D, 89 French Street, New Brunswick, NJ, 08901, USA. ld473@rwjms.rutgers.edu.

ABSTRACT

Background: Alcohol abuse produces an enormous impact on health, society, and the economy. Currently, there are very limited therapies available, largely due to the poor understanding of mechanisms underlying alcohol use disorders (AUDs) in humans. Oxidative damage of mitochondria and cellular proteins aggravates the progression of neuroinflammation and neurological disorders initiated by alcohol abuse.

Results: Here we show that ethanol exposure causes neuroinflammation in both human induced pluripotent stem (iPS) cells and human neural progenitor cells (NPCs). Ethanol exposure for 24 hours or 7 days does not affect the proliferation of iPS cells and NPCs, but primes an innate immune-like response by activating the NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway. This leads to an increase of microtubule-associated protein 1A/1B-light chain 3(+) (LC3B(+)) autophagic puncta and impairment of the mitochondrial and lysosomal distribution. In addition, a decrease of mature neurons derived from differentiating NPCs is evident in ethanol pre-exposed compared to control NPCs. Moreover, a second insult of a pro-inflammatory factor in addition to ethanol preexposure enhances innate cellular inflammation in human iPS cells.

Conclusions: This study provides strong evidence that neuronal inflammation contributes to the pathophysiology of AUDs through the activation of the inflammasome pathway in human cellular models.

No MeSH data available.


Related in: MedlinePlus