Therapeutic Role of a Cysteine Precursor, OTC, in Ischemic Stroke Is Mediated by Improved Proteostasis in Mice

Li, Yanying; Min, Jia-Wei; Feng, Shelley; Subedi, Kalpana; Qiao, Fangfang; Mammenga, Emily; Callegari, Eduardo; Wang, Hongmin

doi:10.1007/s12975-019-00707-w

Cited by 35 publications

(30 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Electrophoresis, transfer, and Western blot analysis of proteins were performed according to previously described methods [ 28 ]. The primary antibodies used were anti-CryAB (1:1000, Abcam; #ab13496), anti-Argonaute 1 (Argo1) (1:1000, Cell Signaling Technology, #5053, Lot 1), anti-Tsg101 (1:1000, Santa Cruz Biotechnology, #sc7964, Lot F0217), anti-ubiquitin (Ub) (1:1000, Cell Signaling Technology, #3936, Lot 14), anti-IL-1β (1:1000, Cell Signaling Technology, #63124), anti-IL-6 (1:1000, Cell Signaling Technology, #12912), anti-TNFα (1:1000, Cell Signaling Technology, #11948), and anti-β-actin (1:1000, Santa Cruz Biotechnology, #sc-1616, Lot G1615).…”

Section: Methodsmentioning

confidence: 99%

Peripherally misfolded proteins exacerbate ischemic stroke-induced neuroinflammation and brain injury

Subedi

Baride

et al. 2021

J Neuroinflammation

Self Cite

View full text Add to dashboard Cite

Background Protein aggregates can be found in peripheral organs, such as the heart, kidney, and pancreas, but little is known about the impact of peripherally misfolded proteins on neuroinflammation and brain functional recovery following ischemic stroke. Methods Here, we studied the ischemia/reperfusion (I/R) induced brain injury in mice with cardiomyocyte-restricted overexpression of a missense (R120G) mutant small heat shock protein, αB-crystallin (CryABR120G), by examining neuroinflammation and brain functional recovery following I/R in comparison to their non-transgenic (Ntg) littermates. To understand how peripherally misfolded proteins influence brain functionality, exosomes were isolated from CryABR120G and Ntg mouse blood and were used to treat wild-type (WT) mice and primary cortical neuron-glia mix cultures. Additionally, isolated protein aggregates from the brain following I/R were isolated and subjected to mass-spectrometric analysis to assess whether the aggregates contained the mutant protein, CryABR120G. To determine whether the CryABR120G misfolding can self-propagate, a misfolded protein seeding assay was performed in cell cultures. Results Our results showed that CryABR120G mice exhibited dramatically increased infarct volume, delayed brain functional recovery, and enhanced neuroinflammation and protein aggregation in the brain following I/R when compared to the Ntg mice. Intriguingly, mass-spectrometric analysis of the protein aggregates isolated from CryABR120G mouse brains confirmed presence of the mutant CryABR120G protein in the brain. Importantly, intravenous administration of WT mice with the exosomes isolated from CryABR120G mouse blood exacerbated I/R-induced cerebral injury in WT mice. Moreover, incubation of the CryABR120G mouse exosomes with primary neuronal cultures induced pronounced protein aggregation. Transduction of CryABR120G aggregate seeds into cell cultures caused normal CryAB proteins to undergo dramatic aggregation and form large aggregates, suggesting self-propagation of CryABR120G misfolding in cells. Conclusions These results suggest that peripherally misfolded proteins in the heart remotely enhance neuroinflammation and exacerbate brain injury following I/R likely through exosomes, which may represent an underappreciated mechanism underlying heart-brain crosstalk.

show abstract

Section: Methodsmentioning

confidence: 99%

Peripherally misfolded proteins exacerbate ischemic stroke-induced neuroinflammation and brain injury

Subedi

Baride

et al. 2021

J Neuroinflammation

Self Cite

View full text Add to dashboard Cite

show abstract

“…Electrophoresis, transfer and western blot analysis of proteins were according to previously described methods 43 . The primary antibodies used were anti-CryAB (1:1000, Abcam; #ab13496), anti-proteasome 20S beta 1i subunit (1:1000, Enzo life sciences, #BML-PW8205; Lot 10071464), anti-proteasome 20S beta 2i subunit (1:1000, Enzo life sciences, #BML-PW8350; Lot 11281405), anti-proteasome 20S beta 5i subunit (1:1000, Cell Signaling Technology, #13726; Lot 1), anti-Argonaute 1 (Argo1) (1:1000, Cell Signaling Technology, #5053, Lot 1), anti-Tsg101 (1:1000, Santa Cruz Biotechnology, #sc7964, Lot F0217), anti-ubiquitin (Ub) (1:1000, Cell Signaling Technology, #3936, Lot 14), anti-K48-linkage specific polyubiquitin (1:1000, Cell Signaling Technology, #8081; Lot 2), and anti-actin (1:1000, Santa Cruz Biotechnology, #sc-1616, Lot G1615).…”

Section: Methodsmentioning

confidence: 99%

“…Primary mouse cortical neurons were prepared from wild-type C57BL/6J mice as previously described 43 . The cells were cultured in poly-ornithine-coated 12-well plates for 7 days and then treated with exosomes (10 µg/ml) isolated either from the Ntg or CryAB R120G mouse blood and after 24 hours, the exosome-treated neuronal cultures were stained for protein aggregates using a Proteostat Aggresome Detection Kit (Enzo Life Sciences, #ENZ-51035) by following the manufacturer provided protocol.…”

Section: Methodsmentioning

confidence: 99%

Peripherally expressed misfolded proteins remotely disrupt brain function and aggravate stroke-induced brain injury

Subedi

Baride

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

Impaired proteostasis has been linked to various diseases, whereas little is known about the impact of peripherally misfolded proteins on the brain. We here studied the brain of mice with cardiomyocyte-restricted overexpression of a missense (R120G) mutant small heat shock protein, α B-crystallin (CryAB R120G ). At baseline, the CryAB R120G mice showed impaired cognitive and motor functions, aberrant protein aggregates, neuroinflammation, impaired bloodbrain barrier, and reduced proteasome activity in the brain compared with their non-transgenic (Ntg) littermates. Ischemic stroke dramatically exacerbated these pathological alterations and caused more severe brain dysfunction in CryAB R120G mice than in the Ntg mice. Intravenously injecting the exosomes isolated from CryAB R120G mouse blood into wild-type mice caused the similar phenotypes seen from CryAB R120G mice. Importantly, the CryAB R120G protein showed the prion-like properties. These results suggest that peripherally misfolded proteins in the heart remotely disrupt brain function through prion-like neuropathology, which may represent an underappreciated mechanism underlying heart-brain crosstalk.

show abstract

“…GSH is an important antioxidant synthesized in cells, which detoxifies ROS. However, the therapeutic effect of OTC diminishes in Ubqln1 knockout mice after cerebral stroke, suggesting that the therapeutic potential of OTC involves both antioxidative property and proteostasis enhancement [113]. Another study from the same group also demonstrated that the administration of nialamide (a non-selective monoamine oxidase inhibitor) in the ischemic stroke mouse model can also upregulate Ubqln1, and in turn reduces oxidative stress as well as neuroinflammation ( Figure 6) [114].…”

Section: Er Stress Components and Ich Therapeutic Strategiesmentioning

confidence: 98%

A Role for Endoplasmic Reticulum Stress in Intracerebral Hemorrhage

Thangameeran

Tsai

Hung

et al. 2020

Cells

View full text Add to dashboard Cite

The endoplasmic reticulum (ER) is an intracellular organelle that performs multiple functions, such as lipid biosynthesis, protein folding, and maintaining intracellular calcium homeostasis. Thus, conditions wherein the ER is unable to fold proteins is defined as ER stress, and an inbuilt quality control mechanism, called the unfolded protein response (UPR), is activated during ER stress, which serves as a recovery system that inhibits protein synthesis. Further, based on the severity of ER stress, the response could involve both proapoptotic and antiapoptotic phases. Intracerebral hemorrhage (ICH) is the second most common subtype of cerebral stroke and many lines of evidence have suggested a role for the ER in major neurological disorders. The injury mechanism during ICH includes hematoma formation, which in turn leads to inflammation, elevated intracranial pressure, and edema. a proper understanding of the injury mechanism(s) is required to effectively treat ICH and closing the gap between our current understanding of ER stress mechanisms and ICH injury can lead to valuable advances in the clinical management of ICH.Cells 2020, 9, 750 2 of 20 because of cardiovascular conditions, with hypertension playing a major role. PBI is characterized by mechanical injury followed by a mass effect with the initial ictus causing physical tissue disruption that then leads to pathophysiological conditions in the brain.A sudden rise in intracranial hematoma volume causes an increase in barotrauma and reduces blood flow to the area of the ictus [4,5]. Typically, PBI is followed by SBI, which is considered as a devastating stage after ICH, and the severity of SBI depends on the rate of recovery and location of the ICH. SBI involves the neuroinflammatory response to the triggering of the coagulation cascade through activation of the immune system. The inflammation size is based on the volume and position of the hematoma [5][6][7]. The various pathological factors responsible for SBI include the host immune response, release of thrombin, release of clot components (iron and heme)

show abstract

Therapeutic Role of a Cysteine Precursor, OTC, in Ischemic Stroke Is Mediated by Improved Proteostasis in Mice

Cited by 35 publications

References 46 publications

Peripherally misfolded proteins exacerbate ischemic stroke-induced neuroinflammation and brain injury

Peripherally misfolded proteins exacerbate ischemic stroke-induced neuroinflammation and brain injury

Peripherally expressed misfolded proteins remotely disrupt brain function and aggravate stroke-induced brain injury

A Role for Endoplasmic Reticulum Stress in Intracerebral Hemorrhage

Contact Info

Product

Resources

About