MicroRNAs modulate CaMKIIα/SIRT1 signaling pathway as a biomarker of cognitive ability in adolescents

Abstract

The dynamic regulation of synaptic plasticity underlies memory formation, involving intricate signaling pathways with both facilitatory and inhibitory roles. MicroRNAs are emerging modulators of memory processes through their fine-tuning of gene expression. To explore the influence of miRNAs on adolescent cognitive function, we investigated the association between academic performance, cognitive ability as measured by the Inquiry for Scientific Thinking, Analytics, and Reasoning test, and plasma miRNA profiling in 486 senior high school students. Our analysis identified 38 differentially expressed miRNAs between students with high and low academic performance. Notably, miR-219 b/548e/628/885 and miR-30a/30c-1/195/204 potentially targeted genes associated with the CaMKII/SIRT1 signaling pathway, a crucial facilitator of memory consolidation. Collectively, our findings suggest that specific plasma miRNAs, particularly the CaMKII/SIRT1-related miR-30a/30c-1/195/204 cluster, potentially serve as promising biomarkers for cognitive function in adolescents. Our findings further support the proposed interaction between NF-kB activity and CaMKIIα in regulating synaptic plasticity. Under hypomethylation conditions, increased NF-kB activity, a key component of inflammation and neural plasticity, influences learning and memory. This biological pathway, representing the initiation of epigenetic memory, demonstrates significant predictive power for both cognitive ability and academic performance.

Keywords: Adolescents; CaMKIIα/SIRT1 signaling pathway; Cognitive ability; NF-kB; miRNA.

Fig. 1

Differential expressed miRNAs assessed by three tests. This figure depicts a Venn diagram illustrating the overlap of differentially expressed miRNAs identified by three independent tests. The circles represent the sets of differentially expressed miRNAs identified by each test. Scientific Reasoning Ability in (SCIA) (light yellow); Comprehensive Assessment program (light blue) and iSTAR (yellow green); Male (Green) and Female (Red). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 2

Phosphorylation of pMeCP2 and pCREB is consistent with the expression of pCaMKIIα. Total soluble protein from HEK293-derived cells were harvested at 2, 4, and 6 d after the induction of CaMKIIα expression with Dox. (a) Representative immunoblots displaying CaMKIIα, phosphor-CaMKIIα(T286), pCREB, MeCP2 and phosphs-MeCP2. H3.3 B was considered a loading control. (b, c) Quantification of relative protein expression levels. Data are presented as mean ± SD values. ∗P < 0.05, n = 5, Student's t-test.

Fig. 3

CaMKIIα phosphorylation increasing the acetylation of H3K9ac and inhibiting the expression of SIRT1. Total soluble protein from HEK293-derived cells were harvested at 2, 4, and 6 d after the induction of CaMKIIα expression with Dox. (a) Representative immunoblots displaying CaMKIIα, phosphor-CaMKIIα(T286), SIRT1, BDNF, and H3K9ac expression. α-Tubulin was considered loading control. (b, c) Quantification of relative protein expression levels. Data are presented as mean ± SD values. ∗P < 0.05, n = 5, Student's t-test.

Fig. 4

Overexpression of miRNAs inducing pro-inflammatory NF-κB and acetylated H3K9. (a) Representative immunoblotting showing the expression of CaMKIIα, and pCaMKIIα, in HEK293 cells transfected with the miR-548e, miR-885, and miRNA-291b-1, and -2) 4 days after the induction of CaMKIIα by Dox. Total soluble protein from HEK293-derived cells was harvested at day 6; (b, c) The relative protein levels were normalized to α-tubulin protein levels. The graph represents the average of three independent experiments with error bars indicating standard deviation (SD; mean ± SD). Statistical significance was determined using Student's t-test (∗p < 0.05). The predicated binding sites for miR-548e/885 and miR-219 b within the 3′ UTR of CaMKIIα were identified using TargetScan.

Fig. 5

Research framework. The diagram depicts the role of Histone modifications in memory formation. Histone modifications, such as methylation and acetylation, lead to chromatin remodeling. In responding to calcium influx through synaptic NMDA receptors triggers long-term potentiation (LTP). Acetylated histones loosen the promoters of genes crucial for memory formation, making it more accessible for transcription machinery and increase genes expression. MicroRNAs that target CaMKIIα and SIRT1 are likely to be involved in the interplay between LTP and LTD for memory consolidation by affecting the balance of the processes. Quantitative and qualitative cognitive ability assessments will be supported by iSTAR test, which emphasize three areas of core skills that support scientific inquiry, including control of variables (COV), data analytics (DA), and causal decision making (CDM).

Fig. 6

A plausible role of microRNAs, NF-kB and SIRT1 in regulating synaptic plasticity by targeting SIRT1, CaMKIIα, and BDNF. Neuronal activity leads to an influx of calcium (Ca²⁺) ions into the postsynaptic neuron. Ca²⁺ binds to calmodulin (CaM), which in turn activates CaMKIIα by inducing a conformational change. This CaM-Ca²⁺-dependent activation allows CaMKIIα to autophosphorylate at a threonine-286), creating an autonomous and persisted CaM-independent activity state which actives GluA1 at excitatory synapses and MeCP2-CREB-BDNF signaling cascades. The negative regulation under hypomethylation conditions, we identified hypomethylation and elevated NF-κB activity, decreased SIRT1, and downregulation of CaMKIIα expression. We postulate that CaMKIIα-mediated synaptic plasticity can be modulated by the synergistic action of miRNAs (particularly miR-30a/30c-1/195/204), NF-kB and SiRT1.