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CaMKIIδ, Stabilized by RNA N6-Methyladenosine Reader IGF2BP2, Boosts Coxsackievirus B3-Induced Myocardial Inflammation via Interacting with TIRAP

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Abstract

Calcium/calmodulin-dependent protein kinase II (CaMKII) has been demonstrated to be aberrantly activated in viral myocarditis (VMC), but the role of its subtype CaMKIIδ in VMC remains unclear.

VMC mice and cardiomyocytes models were induced by Coxsackievirus B3 (CVB3) treatment. Mice that underwent sham surgery and saline-treated cardiomyocytes served as controls. Body weight, survival, left ventricular ejection fraction (LVEF), and fractional shortening (LVFS) were measured, and HE staining was performed to evaluate heart function in VMC mice model and sham control. Inflammation factors in serum or cell supernatant were detected by ELISA. Expressions of CaMKIIδ, Toll/interleukin-1 receptor domain containing adaptor protein (TIRAP), insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2), nuclear factor NF-kappaB (NF-κB) signals, and inflammation factors were examined by quantitative real time polymerase chain reaction (qRT-PCR) or western blot. CCK-8, EdU, and flow cytometry were used to evaluate cell behaviors. Co-immunoprecipitation (Co-IP), RNA immunoprecipitation (RIP), and RNA pull-down were utilized to validate molecule interaction. Methylated RNA immunoprecipitation (MeRIP) was performed to measure N6-methyladenosine (m6A) level of specific molecule.

CaMKIIδ was upregulated in VMC mice and CVB3-treated primary cardiomyocytes, of which knockdown improved cell viability, proliferation, and suppressed cell apoptosis in vitro, thereby alleviating myocarditis in vivo. The stability of CaMKIIδ was attributed to the presence of IGF2BP2 through m6A modification. Loss of CaMKIIδ repressed NF-κB pathway via negatively and directly regulating TIRAP to be involved in inflammatory damage.

CaMKIIδ, stabilized by m6A reader IGF2BP2, modulated NF-κB pathway via interacting with TIRAP to alter cell viability, proliferation, and apoptosis, thereby affecting VMC outcome.

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Data Availability

All data collected and analyzed during the current study are available from the corresponding author upon reasonable request.

Abbreviations

VMC:

viral myocarditis

CaMKII:

calcium/calmodulin-dependent protein kinase II

CVB3:

Coxsackievirus B3

LVEF:

left ventricular ejection fraction

LVFS:

left ventricular fractional shortening

TIRAP:

Toll/interleukin-1 receptor domain containing adaptor protein (TIRAP)

IGF2BP2:

insulin-like growth factor 2 mRNA binding protein 2

NF-κB:

nuclear factor NF-kappaB

qRT-PCR:

quantitative real-time polymerase chain reaction

Co-IP:

co-immunoprecipitation

RIP:

RNA immunoprecipitation

MeRIP:

methylated RNA immunoprecipitation

m6A:

N6-methyladenosine

AMI:

acute myocardial infarction

STAT3:

signal transducer and activator of transcription 3

IL:

interleukin

PBS:

phosphate-buffered saline

METTL3:

methyltransferase 3

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Funding

The work was supported by Natural Science Foundation of Jiangxi Province (No. 20232BAB206015).

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Authors and Affiliations

  1. Department of Respiratory Medicine, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, 330006, Jiangxi Province, People’s Republic of China

    Qingping Xiao

  2. Department of Cardiology, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, No. 17, Yongwaizheng Street, Donghu District, Nanchang, 330006, Jiangxi Province, People’s Republic of China

    Lijuan Liu, Wei Qian, Ting Kang, Ru Ying & Jungang Nie

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  1. Qingping Xiao
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  2. Lijuan Liu
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  3. Wei Qian
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  5. Ru Ying
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  6. Jungang Nie
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Contributions

Conception and design of study: JN; Acquisition of data: QX, WQ, RY; Analysis and interpretation of data: LL, TK; Drafting the manuscript: QX; Revising the manuscript critically for important intellectual content: JN.

Corresponding author

Correspondence to Jungang Nie.

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Ethics Approval

All the animal procedures were performed in accordance with the Guiding Principles in the Use and Care of Animals and approved by the Ethical Committee of The First Affiliated Hospital of Nanchang University.

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The authors declare no competing interests.

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Associate Editor Joost Sluijter oversaw the review of this article

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Highlights

1. Knockdown of CaMKIIδ alleviates apoptosis and inflammation of cardiomyocytes in the cell model of VMC.

2. IGF2BP2 stabilizes CaMKIIδ by m6A modification.

3. CaMKIIδ interacts with and positively regulates TIRAP.

4. Knockdown of CaMKIIδ alleviates VMC through TIRAP inhibition in vivo.

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Xiao, Q., Liu, L., Qian, W. et al. CaMKIIδ, Stabilized by RNA N6-Methyladenosine Reader IGF2BP2, Boosts Coxsackievirus B3-Induced Myocardial Inflammation via Interacting with TIRAP. J. of Cardiovasc. Trans. Res. (2024). https://doi.org/10.1007/s12265-023-10478-3

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