Verapamil Inhibits Ser202/Thr205 Phosphorylation of Tau by Blocking TXNIP/ROS/p38 MAPK Pathway

Melone, Mariarosa Anna Beatrice; Dato, Clemente; Paladino, Simona; Coppola, Cinzia; Trebini, Claudia; Giordana, Maria Teresa; Perrone, Lorena

doi:10.1007/s11095-017-2276-2

Cited by 50 publications

(36 citation statements)

References 41 publications

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“…Forty-eight hours later, cells were harvested and processed for TXNIP expression by western blot analysis. Images were analyzed with the software Zen (Zeiss) as previously described (14,19,21).…”

Section: Lentiviral Tools and Lentiviral Transduction In Primary Micrmentioning

confidence: 99%

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Rage-txnip Axis Drives Inflammation in Alzheimer`s by Targeting aβ to Mitochondria in Microglia

Perrone

Abate

Uberti

et al. 2020

Preprint

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Alzheimer’s disease (AD) is the most common form of dementia characterized by progressive memory loss and cognitive decline. Although neuroinflammation and oxidative stress are well-recognized features of AD, their correlations with the early molecular events characterizing the pathology are not yet well clarified. Here we demonstrated the role of RAGE-TXNIP axis in neuroinflammation in relation to amyloid-beta (Aβ) burden in both invivo and in-vitro models. In the hippocampus of 5xFAD mice, microglial activation correlates with increased TXNIP expression, and TXNIP silencing or its pharmacological inhibition prevented the neuroinflammation in those mice. TXNIP increased expression has been found also in human subjects affected by AD and other dementia when plasma samples were analyzed, suggesting TXNIP as a putative biomarker of neurodegeneration and cognitive decline. To better elucidate whether a mechanistic link between RAGE-TXNIP axis and AD exist, we investigated how TXNIP is involved in Aβ intracellular trafficking downstream RAGE. Indeed, TXNIP is an α-arrestin protein involved in endocytosis. We found TXNIP associated to RAGE and Aβ. RAGE-TXNIP axis was required for targeting Aβ into mitochondria, as shown in primed primary microglia exposed to Aβ dimers. Silencing of TXNIP or inhibition of RAGE activation reduced Aβ transport from the cellular surface to mitochondria. Detrimental effect of Aβ in mitochondria has been also confirmed by evaluating increased mitochondrial- and intracellular- oxidative stress. When RAGE-TXNIP axis is down-regulated, mitochondrial functionality was restored and Aβ toxicity was mitigated. Furthermore, Aβ transport into mitochondria and subsequent mitochondrial dysfunction drove inflammation in microglia cells via NLRP3 inflammasome activation, and down-regulation of RAGE-TXNIP axis was able to efficiently mitigate Aβ proinflammatory effect. Overall this data suggest RAGE-TXNIP axis is required for NLRP3 inflammasome activation induced by Aβ in microglia.Our work shed light on novel mechanism of action of RAGE-TXNIP axis in microglia, which is intertwined with Aβ and ultimately causes inflammation, suggesting TXNIP as a druggable target to be better deepened for AD.

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Section: Lentiviral Tools and Lentiviral Transduction In Primary Micrmentioning

confidence: 99%

“…The measurement of IL-1β in extracts from mice hippocampus was performed using the same ELISA kit according to the manufacturer's instruction (BD Biosciences). Protein extracts from mice hippocampus were obtained as previously described (14).…”

Section: Measurement Of Secreted Il-1β and Caspase-1 Activitymentioning

confidence: 99%

Rage-txnip Axis Drives Inflammation in Alzheimer`s by Targeting aβ to Mitochondria in Microglia

Perrone

Abate

Uberti

et al. 2020

Preprint

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“…Verapamil (Sigma, USA) was incorporated into animals' drinking water by a blinded investigator; and adjusted to dose them with 1 mg/kg/day verapamil for 1 month. Verapamil dosage was selected based on the lowest required for TXNIP suppression [17], as addressed in earlier reports without affecting blood pressure (Suppl. Figure 3).…”

Section: Animalsmentioning

confidence: 99%

“…Although no signi cant change were detected by ANOVA, TRX showed tangible upregulation in TXNIP −/− brains compared to wild type aged brains (P = 0.0007), using T-test. For pharmacological modulation of TXNIP, we dosed aged animals with verapamil, an L-type Ca ++ channel blocker, that is well known to suppress TXNIP expression [17]. At this particular dosage (1 mg/kg/day, 1 month), far too low to reduce blood pressure (Suppl.…”

Section: Genetic and Pharmacological Inhibition Of Txnip Prevents Agementioning

confidence: 99%

TXNIP regulates age-associated neuroinflammation: Implication in Alzheimer’s disease

Ismael¹,

Nasoohi²,

Yoo³

et al. 2020

Preprint

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Background Immune system hypersensitivity with aging is believed to contribute to mental frailty in elderlies. This is postulated to arise from accumulation of oxidative molecular patterns. Solid evidences delineates thioredoxin interacting protein (TXNIP), an inducible protein involved in oxidative stress, is essential for NOD-like receptor pyrin domain containing-3 (NLRP3)-inflammasome activation which intimately connects “inflammaging” to senile cognitive decline. This study aims to fundamentally explore the plausible involvement of TXNIP/NLRP3 inflammasome pathway in senile dementia and the typical Alzhemier’s disease. Methods In experimental studies cerebral samples from gender-matched mice were compared for TXNIP/NLRP3 inflammasome activation and klotho depletion, through immunoblotting and immunostaining in different life span points. In aged males, genetic or pharmacological ablation of TXNIP were then used to determine effects on cognitive decline and sensorimotor frailty in morris water maze, novel object recognition test and gait control analysis. Immunoblotting/staining experiments were also performed on human postmortem aged hippocampal specimens and 5XFAD transgenic mice, to ultimately address Alzheimer’s disease (AD) as the most age related dementia. Results According to our preclinical studies, cerebral TXNIP was significantly upregulated in aged animals, paralleled by the NLRP3-inflammasome over-activity in both sexes, and closely associated klotho depletion in aged males. TXNIP knock-out reversed age-related NLRP3-hyperactivity and enhanced thioredoxin (TRX) levels in aged brains. Further, pharmacological TXNIP inhibition replicated the TXNIP/NLRP3-inflammasome downregulation in aged animals, with FOXO-1 and mTOR upregulation. These alterations concurred with substantial improvements in both cognitive and sensorimotor abilities. Moreover, our immunostaining shows a significant increase of TXNIP in transgenic 5XFAD mice brain and TXNIP/NLRP3-inflammasome activity in AD human postmortem hippocampal specimens, in proximity of p-tau tangles and β-amyloid plaques. Conclusion Together, these findings substantiate the pivotal role of TXNIP to drive inflammging in parallel with klotho depletion and functional decline. TXNIP co-localization with hallmarks of AD pathology is further supportive of potential mechanistic links between TXNIP and AD. Unraveling new information on upstream pathways, these data support modulating thioredoxin system as a potential approach to decelerate senile frailty.

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“…Verapamil is a calcium channel blocker that has been shown to downregulate the expression of the NLRP3 binding protein TXNIP, 18,19 and thus has been used in several studies to assess a role for this critical structural component of the NLRP3 inflammasome in DAMP pathways mediated by ROS. 20,21 As shown in Figure 3A, a dose of 150 μM Verapamil was sufficient to completely inhibit CPPD-mediated caspase-1 activation (IC 50 =100 μM).…”

Section: Verapamil Inhibits Caspase-1 Activationmentioning

confidence: 99%

<p>Calcium Pyrophosphate And Monosodium Urate Activate The NLRP3 Inflammasome Within Bladder Urothelium Via Reactive Oxygen Species And TXNIP</p>

Harper

Leidig

Hughes

et al. 2019

RRU

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Objective: To investigate the in vitro activation of the NLRP3 inflammasome within bladder urothelium by stone-forming components. Further, to describe the contributions of reactive oxygen species (ROS) and thioredoxin-interacting protein (TXNIP), an important structural component of the inflammasome, to this activation. Methods: Urothelial cells were harvested and incubated overnight. For agonist studies, cells were treated with varying concentrations of calcium pyrophosphate (CPPD) and monosodium urate (MSU). For inhibitor studies, cells were treated with either N-acetylcysteine (NAC) (1 hr) or Verapamil (4 hrs) prior to incubation with either CPPD (62.5 ug/mL) or MSU (1.25 ug/mL) for 24 hrs. Untreated controls were incubated with ATP (1.25 mM) for 1 hr to maximally stimulate NLRP3 inflammasome activity (measured as caspase-1 cleavage of the fluorogenic substrate Ac-YVAD-AFC). Results are reported as a percentage of maximum ATP response. Results: CPPD and MSU activate caspase-1 in urothelial cells in a dose-dependent manner, reaching~50% and~25% of the ATP response, respectively. Pre-treatment with the general ROS scavenger NAC reduces this activation in a dose-dependent manner. Additionally, activation was suppressed through treatment with Verapamil, a known downregulator of TXNIP expression. Conclusion: The stone components CPPD and MSU activate NLRP3 in an ROS and TXNIP-dependent manner in bladder urothelium. These findings demonstrate the importance of ROS and TXNIP, and suggest that targeting either may be a way to decrease stonedependent NLRP3 inflammation within the bladder.

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Verapamil Inhibits Ser202/Thr205 Phosphorylation of Tau by Blocking TXNIP/ROS/p38 MAPK Pathway

Abstract: Our study unveil a novel pathway involved in AD progression that is inhibited by Verapamil, shedding new light on the understanding of the therapeutic potential of Verapamil in AD.

Cited by 50 publications

References 41 publications

Rage-txnip Axis Drives Inflammation in Alzheimer`s by Targeting aβ to Mitochondria in Microglia

Rage-txnip Axis Drives Inflammation in Alzheimer`s by Targeting aβ to Mitochondria in Microglia

TXNIP regulates age-associated neuroinflammation: Implication in Alzheimer’s disease

<p>Calcium Pyrophosphate And Monosodium Urate Activate The NLRP3 Inflammasome Within Bladder Urothelium Via Reactive Oxygen Species And TXNIP</p>

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