Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , Schematic workflow of the experiment demonstrating the process of extracting tRNAs from the kidneys of adult and aged mice, followed by digestion with RNase T1 and LC-MS analysis. Proteins were extracted for proteomic analysis. b , Heatmap of tRNA mapping in the kidneys of aged mice compared with adult mice. c , Proteomic analysis of the kidneys of adult and aged mice revealed significant enrichment of pathways such as immune system process and immune response in GO-BP enrichment. d , Volcano plot of selenoproteins level in the kidney from adult and aged mice. e , Enrichment of tRNA half fraction from the kidney of adult and aged mice. f , By comparing the theoretical MS data with the measured data, the RNA sequence corresponding to the ion m/z 2271.4690 was identified as the 5′-tRNA half cleaved at position 34 of tRNA Sec(NCA) . g , The purified RNA was further characterized as 5′-tRNA Sec(NCA) half by RNase T1 digestion and LC-MS/MS analysis. h , 5′-tRNA Sec(NCA) half was particularly detected in humans and mice, but not in C. elegans , D. melanogaster , and yeast. i-j , The 5′-tRNA Sec(NCA) half was significantly accumulated in the kidneys and white blood cells of mice during aging. k , The abundance of 5′-tRNA Sec(NCA) half increased with age in healthy human white blood cell samples. Data presented in i and k are presented as mean ± S.E.M. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001; n.s., non-significant.

Article Snippet: Subsequently, total mRNA were extracted from whole blood by using Magnetic Tissue/Cell/Blood Total RNA Kit (TIANGEN Biotech, China) according to the manufactures’ protocol, followed by RT-qPCR analysis.

Techniques: Liquid Chromatography with Mass Spectroscopy, Sequencing, Purification, Two Tailed Test

Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , CCK-8 assay showed that the proliferation of TCMK-1 and 2BS treated with 5′-tRNA Sec(NCA) half was significantly inhibited compared with the blank transfection reagent group. b . SA-β-gal staining indicated that the SA-β-gal positive rate of TCMK-1 and 2BS cells treated with 5′-tRNA Sec(NCA) half increased significantly, presenting a cellular senescence phenotype. c , The expression levels of senescence markers P21, P16, IL6, and CCL2 were significantly upregulated in cells induced by 5′-tRNA Sec(NCA) half. d , ROS significantly increased in cells treated with 5′-tRNA Sec(NCA) half. e , The telomere length (T/S ratio) of cells treated with 5′-tRNA Sec(NCA) half significantly decreased. f . IFN-β levels significantly upregulated in cells treated with 5′-tRNA Sec(NCA) half, while the level of IFN-γ did not exhibit such huge change. g-h , Liposome nanoparticle (LNP) delivery system encapsulated with 5′-tRNA Sec(NCA) half was intravenously injected into young Balb/c female mice. After 6 days, the senescence markers in the kidneys of mice were significantly upregulated. Data presented in c , d , e , and f are presented as mean ± S.D. Data presented in h are presented as mean ± S.E.M. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001; n.s., non-significant.

Article Snippet: Subsequently, total mRNA were extracted from whole blood by using Magnetic Tissue/Cell/Blood Total RNA Kit (TIANGEN Biotech, China) according to the manufactures’ protocol, followed by RT-qPCR analysis.

Techniques: CCK-8 Assay, Transfection, Staining, Expressing, Injection, Two Tailed Test

Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , Transcriptomic sequencing revealed that dsRNA sensor genes significantly upregulated in cells treated with 5′-tRNA Sec(NCA) half. b , qPCR analysis confirmed that the expression of the above genes and MYD88, a key adaptor protein in the TLR7 signaling pathway, increased significantly. c , In TCMK-1 and 2BS cells treated with 5′-tRNA Sec(NCA) half, the protein level of TLR7 was significantly upregulated in a dose-dependent manner. d , Simulation of the interaction between 5′-tRNA Sec(NCA) half and TLR7 protein predicted by AlphaFold 3. e , RNA pull-down assay demonstrated that 5′-tRNA Sec(NCA) half specifically bound to TLR7. f-g , In TLR7-knockdown (TLR7-KD) cells, co-treatment with 5′-tRNA Sec(NCA) half and siTLR7 led to a significant downregulation of senescence markers and a lower SA-β-gal positive rate compared with treatment with 5′-tRNA Sec(NCA) half alone. Data presented in b , c , and f are presented as mean ± S.D. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001.

Article Snippet: Subsequently, total mRNA were extracted from whole blood by using Magnetic Tissue/Cell/Blood Total RNA Kit (TIANGEN Biotech, China) according to the manufactures’ protocol, followed by RT-qPCR analysis.

Techniques: Sequencing, Expressing, Pull Down Assay, Knockdown, Two Tailed Test

Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , 5′-tRNA Sec(NCA) half significantly increased in D-galactose-induced TCMK-1 aged cells and 2BS naturally aged cells. b , CCK-8 assay indicated that the proliferation of aged cells treated with ASO-5′-tRNA Sec(NCA) half significantly increased compared with the liposome-treated controls. c . Treatment with ASO-5′-tRNA Sec(NCA) half resulted in a significant decrease in the number of SA-β-gal-positive cells in aged cells. d , ROS levels were markedly reduced in aged cells treated with ASO-5′-tRNA Sec(NCA) half. e-f . ASO-5′-tRNA Sec(NCA) half significantly downregulated aging markers and extended telomere length in aged cells. g , LNP delivery of ASO-5′-tRNA Sec(NCA) half significantly extended the lifespan of paraquat-induced acute aged mice in a dose-dependent manner, as well as dysregulating TLR7, P21 and IL6 in multiple organs ( h ). Data presented in a , b , d , e , f and h are presented as mean ± S.D. Data presented in g are presented as log-rank test and the Gehan-Breslow-Wilcoxon. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; *** P <0.001; **** P <0.0001; n.s., non-significant.

Article Snippet: Subsequently, total mRNA were extracted from whole blood by using Magnetic Tissue/Cell/Blood Total RNA Kit (TIANGEN Biotech, China) according to the manufactures’ protocol, followed by RT-qPCR analysis.

Techniques: CCK-8 Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , Experimental design of anti-aging test of ASO-5′-tRNA Sec(NCA) half from 16-month-old mice. b , Image of 20-month-old mice received LNP encapsulated ASO-5′-tRNA Sec(NCA) half treatment or not. Survival curves ( c ) and weight curve ( d ) of aged mice received LNP encapsulated ASO-5′-tRNA Sec(NCA) half treatment (n=20) or not (n=20). e , Bone mineral density measured by 3D X-ray imaging. f-g , Mice treated with ASO-5′-tRNA Sec(NCA) half had a significantly higher lean percent and a lower fatty percent. h , Movement of mice received ASO-5′-tRNA Sec(NCA) half treatment or not in the open-field test. i , Alteration rates of ASO-5′-tRNA Sec(NCA) half-treated mice or not in the Y maze test. Grip strength test ( j ) and rotarod test ( k ) of ASO-5′-tRNA Sec(NCA) half-treated mice or not. l , There was no significant difference in the immobility time between the two groups of mice. m , RRBS sequencing of DNA methylation age showed that 20-month-old aged mice treated with ASO-5′-tRNA Sec(NCA) half were approximately 18.6 weeks younger than the LNP control group. Aging markers expressions ( n ) and telomere length ( o ) from the major organs of ASO-5′-tRNA Sec(NCA) half-treated mice or not. p , H&E staining and IHC staining of P16, P21, and IL6 in major organs from mice treated with ASO-5′-tRNA Sec(NCA) half or not. Data presented in d , e , f , g , h , i , j , k , l , m , n , and o are presented as mean ± S.E.M. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; *** P <0.001; n.s., non-significant. Data presented in c are presented as the log-rank test in orange and the Gehan-Breslow-Wilcoxon in purple.

Article Snippet: Subsequently, total mRNA were extracted from whole blood by using Magnetic Tissue/Cell/Blood Total RNA Kit (TIANGEN Biotech, China) according to the manufactures’ protocol, followed by RT-qPCR analysis.

Techniques: Imaging, Sequencing, DNA Methylation Assay, Control, Staining, Immunohistochemistry, Two Tailed Test

Journal: bioRxiv

Article Title: Targeted intervention of senescence induced by a 5′ half fragment of tRNA Seca(NCA) with antisense oligonucleotide extends healthspan and lifespan in mice

doi: 10.1101/2025.03.24.644848

Figure Lengend Snippet: a , Heatmap of interferon-associated genes in aged cells treated with ASO-5′-tRNA Sec(NCA) half or not by transcriptomic sequencing. b , Volcano plot revealed that BCAT1 dysregulated in ASO-5′-tRNA Sec(NCA) half-treated TCMK-1 cells. c , Putative binding sites of ASO-5′-tRNA Sec(NCA) half on the 3′UTR region of BCAT1 mRNA of mice and humans. d , The dual-luciferase reporter assay demonstrated that ASO-5′-tRNA Sec(NCA) half remarkably decreased the luciferase activity of mice BCAT1-WT reporter vector, while no significant alteration was detected in the luciferase activity of mice BCAT1-MUT reporter gene vector. e , qPCR analysis confirmed that ASO-5′-tRNA Sec(NCA) half significantly inhibited BCAT1 expression in aged cells. f-g , In paraquat-induced mice treated with ASO-5′-tRNA Sec(NCA) half, BCAT1 was downregulated at both the mRNA and protein levels in the kidneys. h , Major organs including liver, spleen, lung, kidney, and muscle from naturally aged mice receiving ASO-5′-tRNA Sec(NCA) half treatment exhibited remarkably lower mRNA levels of BCAT1. i , IHC staining of BCAT1 in major organs from naturally aged mice. Data presented in d and e are presented as mean ± S.D. Data presented in f , g , and h are presented as mean ± S.E.M. Statistical test: two-tailed unpaired t -test. * P <0.05; ** P <0.01; **** P <0.0001; n.s., non-significant.

Article Snippet: Subsequently, total mRNA were extracted from whole blood by using Magnetic Tissue/Cell/Blood Total RNA Kit (TIANGEN Biotech, China) according to the manufactures’ protocol, followed by RT-qPCR analysis.

Techniques: Sequencing, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Plasmid Preparation, Expressing, Immunohistochemistry, Two Tailed Test

Journal: Asia Pacific Allergy

Article Title: Relationship between changes in the 7-day urticaria activity score after treatment with omalizumab and the responsiveness of basophils to FcεRI stimulation in patients with chronic spontaneous urticaria

doi: 10.5415/apallergy.2020.10.e12

Figure Lengend Snippet: Comparison of the serum concentrations of anti-IgE AAbs (A) and anti-FcεRIα AAbs (B), and the FcεRI-crosslinking ability of these AAbs (C, D) among nonatopic control (NC) subjects (n = 20), BAT-negative patients (n = 9), and BAT-positive patients (n = 13). The luciferase expression levels are represented as the fold increase in light units as compared to the results from the nonstimulated cells. Statistical analyses were performed using the Mann-Whitney U test. AAbs, autoantibodies; BAT, basophil activation test. BAT-Neg. Pt., BAT-negative patients; BAT-Pos. Pt., BAT-positive patients.

Article Snippet: Heparinized whole blood was incubated with anti-FcεRI antibody (clone CRA1, BioAcademia, Osaka, Japan; 3 μg/mL), anti-IgE antibody (clone E124-2-8D, Beckman Coulter Inc.; 10 μg/mL), N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) (Sigma-Aldrich, Darmstadt, Germany; 1 μg/mL) or the negative control substance (phosphate-buffer saline), as indicated, for 15 minutes.

Techniques: Comparison, Control, Luciferase, Expressing, MANN-WHITNEY, Activation Assay