Dataset Information

De-2-hydroxyisobutyrylation of clusterin attenuates neuropathology and memory deficits in an Alzheimer's disease mouse model

ABSTRACT: Backgroud Alzheimer's disease (AD) is a progressive and irreversible neurodegenerative disorder for which there is currently no cure, representing a serious clinical challenge. Lysine 2-hydroxyisobutyrylation (Khib) is a newly discovered post-translational modification that has been found to be widespread in a variety of organisms in recent years and is involved in important biometabolic processes that regulate protein function. However, it is unclear whether and how protein Khib modifications are involved in AD. This study aimed to explore the involvement of Khib in AD and its underlying mechanisms. Methods We compared a series of lysine acylation modification patterns between 9-month-old APP/PS1 mice and wild-type (WT) control mice by Western blot assays. Quantitative Khib proteomic analysis identified Khib modification proteins and key sites associated with AD. Using stereotactic brain injection, we delivered adeno- associated viruse (AAV) overexpressing Clusterin (CLU) K436R (lysine to arginine mutation at site 436, mimicking Khib deficiency) into the bilateral lateral ventricles of 9-month-old WT and APP/PS1 mice. Additionally, C8-D1A cells were infected with lentivirus carrying CLU K436 mutation. These approaches enabled functional assessment of CLU K436 khib modification in AD pathogenesis. Furthermore, co-immunoprecipitation analyses revealed that CLU K436 khib modification modulates the interaction between CLU and Aβ. Results The level of Khib was significantly increased in 9-month-old APP/PS1 mice. Quantitative Khib analysis identified Clusterin (CLU) as a differentially modified protein, with K436 serving as its primary Khib modification site. Further investigation revealed that CLU K436 Khib levels was upregulated in APP/PS1 AD model mice. Loss of K436 Khib in CLU K436R mutant APP/PS1 mice notably ameliorated β-amyloid plaque, gliosis, and neuroinflammation, consequently mitigating memory deficits in vivo. The lentiviral vector carrying CLU K436R mutant in C8-D1A astrocyte cell enhanced glial cell activation and cell survival in vitro. Mechanistically, CLU K436 Khib modification may affect Aβ metabolism by impacting the interaction between CLU and Aβ. Conclusions Our results indicated the role of Khib in AD, elucidating the function and potential molecular mechanism of CLU K436 Khib modification in AD. These findings offer novel perspectives on therapeutic approaches for AD.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain

SUBMITTER: Xiaoping Peng

LAB HEAD: Xiaoping Peng

PROVIDER: PXD066261 | Pride | 2026-03-04

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	KA212LP2o_A1_Slot1-41_1_2142.d.zip	Other
	KA212LP2o_A2_Slot1-42_1_2144.d.zip	Other
	KA212LP2o_A3_Slot1-43_1_2146.d.zip	Other
	KA212LP2o_C1_Slot1-38_1_2136.d.zip	Other
	KA212LP2o_C2_Slot1-39_1_2138.d.zip	Other

Items per page:

1 - 5 of 15

Similar Datasets

Project description:Astrocytes, one of the most resilient cells in the brain, transform into reactive astrocytes in response to toxic proteins such as amyloid beta (Aβ) in Alzheimer’s disease (AD). However, reactive astrocyte-mediated non-cell autonomous neuropathological mechanism is not fully understood yet. We aimed our study to find out whether Aβ-induced proteotoxic stress affects the expression of autophagy genes and the modulation of autophagic flux in astrocytes, and if yes, how Aβ-induced autophagy-associated genes are involved Aβ clearance in astrocytes of animal model of AD. Whole RNA sequencing (RNA-seq) was performed to detect gene expression patterns in Aβ-treated human astrocytes in a time-dependent manner. To verify the role of astrocytic autophagy in an AD mouse model, we developed AAVs expressing shRNAs for MAP1LC3B/LC3B (LC3B) and Sequestosome1 (SQSTM1) based on AAV-R-CREon vector, which is a Cre recombinase-dependent gene-silencing system. Also, the effect of astrocyte-specific overexpression of LC3B on the neuropathology in AD (APP/PS1) mice was determined. Neuropathological alterations of AD mice with astrocytic autophagy dysfunction were observed by confocal microscopy and transmission electron microscope (TEM). Behavioral changes of mice were examined through novel object recognition test (NOR) and novel object place recognition test (NOPR). Here, we show that astrocytes, unlike neurons, undergo plastic changes in autophagic processes to remove Aβ. Aβ transiently induces expression of LC3B gene and turns on a prolonged transcription of SQSTM1 gene. The Aβ-induced astrocytic autophagy accelerates urea cycle and putrescine degradation pathway. Pharmacological inhibition of autophagy exacerbates mitochondrial dysfunction and oxidative stress in astrocytes. Astrocyte-specific knockdown of LC3B and SQSTM1 significantly increases Aβ plaque formation and hypertrophic reactive astrocytes in APP/PS1 mice, along with a significant reduction of neuronal marker and cognitive function. In contrast, astrocyte-specific overexpression of LC3B reduced Ab aggregates in the brain of APP/PS1 mice An increase of LC3B and SQSTM1 protein is found in astrocytes of the hippocampus in AD patients. Taken together, our data indicates that Aβ-induced astrocytic autophagic plasticity is an important cellular event to modulate Aβ clearance and maintain cognitive function in AD mice.

Dataset Information

De-2-hydroxyisobutyrylation of clusterin attenuates neuropathology and memory deficits in an Alzheimer's disease mouse model

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets