Journal: bioRxiv

Article Title: Evolutionarily Conserved Decline of tRNA Mannosyl-Queuosine Links Translational Regulation to Aging and Is Reversed by Queuine

doi: 10.64898/2026.03.22.713446

Figure Lengend Snippet: Age-associated queuine depletion correlates with cellular senescence. (A) Age-dependent decline of serum queuine in rats. Queuine concentrations in male rats at 6, 24, and 36 months (n = 6/group; *** p < 0.0001 by linear regression). (B) Age-dependent decline of plasma queuine in humans. Queuine levels across age cohorts: 30-40, 40-50, 50-60, and ≥60 years (n = 25/group; *** p < 0.0001 by linear regression). (C) Senescence-associated β -galactosidase (SA- β -gal) activity. Representative images of SA- β -gal staining (blue) in HEK 293T cells cultured in horse serum (HS) vs. fetal bovine serum (FBS) for 72 hr (scale bar: 50 μm). (D) Senescence marker expression. Relative mRNA levels of p16 and p21 in HS-vs. FBS-cultured cells (n = 3; * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Queuine hydrochloride (CAS: 69565-92-0, TRC-Q525000-0.5MG), was purchased from Toronto Research Chemicals.

Techniques: Clinical Proteomics, Activity Assay, Staining, Cell Culture, Marker, Expressing

Journal: bioRxiv

Article Title: Evolutionarily Conserved Decline of tRNA Mannosyl-Queuosine Links Translational Regulation to Aging and Is Reversed by Queuine

doi: 10.64898/2026.03.22.713446

Figure Lengend Snippet: Queuine supplementation attenuates senescence and extends survival across models. (A) Experimental design: Queuine rescue in HS-cultured 2BS cells. Schematic of early-passage (P30) cells treated with: FBS (control), HS (queuine-deficient), or HS + 10 ng/mL queuine (HS+Q). (B) Senescence marker reversal. p16 and p21 mRNA in HS-cultured 2BS cells ± queuine rescue (n=3; ** p <0.01 vs. HS). (C) Experimental design: Queuine in natural aging. 2BS cells (P30) cultured continuously in FBS ± 10 ng/mL queuine. (D) Natural senescence attenuation. Senescence markers in P39 2BS ± queuine (n=3; ** p <0.01 vs. FBS). (E) Dose-response design. 2BS cells treated with 0/10/50 ng/mL queuine in FBS. (F) Dose-dependent effects. Marker expression at different doses (n=3; ** p <0.01 vs. 0 ng/mL). (G) D. melanogaster supplementation protocol. Queuine (50 ng/mL) added to food. (H) D. melanogaster lifespan extension. Survival curve of queuine treatment and control flies (n=200; log-rank p <0.001). (I) Acute mouse aging protocol. Paraquat (50 mg/kg) ± queuine (25/200 μg/kg) in 6-month C57BL/6J males. (J) Acute survival rescue. Percent survival post-paraquat ± queuine (n=6/group; ** p <0.01). (K) Senescence markers. p16 and p21 mRNA in acute model (n=3; * p <0.05 vs. paraquat-only). (L) Chronic mouse aging protocol. Queuine (10 μg/kg) administered orally to 16-month C57BL/6J males. (M) Longevity extension. Kaplan-Meier curve of naturally aged mice (n=30; log-rank p <0.001).

Article Snippet: Queuine hydrochloride (CAS: 69565-92-0, TRC-Q525000-0.5MG), was purchased from Toronto Research Chemicals.

Techniques: Cell Culture, Control, Marker, Expressing

Journal: bioRxiv

Article Title: Evolutionarily Conserved Decline of tRNA Mannosyl-Queuosine Links Translational Regulation to Aging and Is Reversed by Queuine

doi: 10.64898/2026.03.22.713446

Figure Lengend Snippet: Queuine supplementation extends healthspan in D. melanogaster . (A) Schematic of experimental design assessing queuine-mediated healthspan improvement in D. melanogaster . Wild-type flies and flies supplemented with queuine (50 ng per food bottle) were evaluated for weight, exercise capacity, heat stress resistance, olfactory memory, and antioxidant capacity at 10, 20, and 30 days post-eclosion. (B-G) Longitudinal analysis of healthspan parameters in control and queuine-supplemented flies (n=10 cohorts): body weight (B), exercise performance (D), heat stress survival (E), olfactory memory retention (F), and systemic antioxidant activity (G).

Article Snippet: Queuine hydrochloride (CAS: 69565-92-0, TRC-Q525000-0.5MG), was purchased from Toronto Research Chemicals.

Techniques: Olfactory, Control, Antioxidant Activity Assay

Journal: bioRxiv

Article Title: Evolutionarily Conserved Decline of tRNA Mannosyl-Queuosine Links Translational Regulation to Aging and Is Reversed by Queuine

doi: 10.64898/2026.03.22.713446

Figure Lengend Snippet: Multisystem rejuvenation through queuine supplementation in naturally aging mice. (A) Experimental design: Middle-aged (16-month-old) wild-type male C57BL/6J mice received oral queuine (10 μg/kg bw/day; n=6) or vehicle once every three days. Aging hallmarks, behavioral/cognitive function, molecular biomarkers (telomerase activity, DNA methylation clock, inflammatory cytokines, antioxidant capacity), clinical biochemistry (hematology, liver/kidney function, glucose/lipids, myocardial enzymes), and gut microbiota were assessed at 8, 24, and 40 weeks post-treatment. (B-P) Queuine-mediated improvements in: Molecular aging biomarkers: p16/p21 expression (B), telomerase activity (C), serum IL-6 (D), systemic antioxidant capacity (E). Behavior/cognition: Open Field locomotion (F), physical appearance (G), Novel Object Recognition (H-left) and Object Location Memory (H-right), endurance (I). Epigenetic aging: DNA methylation age (J). Clinical pathology: Hematological indices (K), liver transaminases (L), fasting glucose (M), lipid profile (N), renal biomarkers (O), and cardiac enzymes (P). (Q-T) Gut microbiota modulation by queuine: (Q) Venn diagram of core microbial OTUs in young (6mo), aged (22mo), and aged+queuine (22mo) mice. (R) Phylum-level taxonomic composition, (S) Species significantly enriched in aged+queuine vs. aged mice. (T) Species significantly enriched in aged vs. young mice.

Article Snippet: Queuine hydrochloride (CAS: 69565-92-0, TRC-Q525000-0.5MG), was purchased from Toronto Research Chemicals.

Techniques: Activity Assay, DNA Methylation Assay, Expressing

Journal: bioRxiv

Article Title: Evolutionarily Conserved Decline of tRNA Mannosyl-Queuosine Links Translational Regulation to Aging and Is Reversed by Queuine

doi: 10.64898/2026.03.22.713446

Figure Lengend Snippet: Queuine-dependent tRNA modification manQ deficiency emerges as a conserved epitranscriptomic hallmark of aging. (A) Quality control of rat kidney RNA isolation. Top: Urea-PAGE of large (lanes 1-4) and small RNA (lanes 7-10); Ladders (lane 5: Low range, lane 6: miRNA). Bottom: Small RNA UV spectra (Rat-6M, 2M, 36M, 24M). (B) OPLS-DA score plot of tRNA fragments across age groups (t[1] = 0.527, t[2] = 0.066). (C-D) Age-dependent tRNA fragment alterations: Heatmap (C) and volcano plot (D) comparing 6mo vs 36mo rats. (E) Structural validation of CUC[manQ]UCA[m 5 C]G fragment by UHPLC-QTOF-MS/MS. CID spectrum shows c/y-ion series (mass tolerance ±0.5 Da). (F) Top: EIC chromatograms and quantitation of CUC[manQ]UCA[m 5 C]G fragment (*** p <0.001 vs 6mo). Bottom: Control fragment UAUCCCG (ns). All data normalized to TΨCG peak area. (G) Purification of tRNAᴬˢᵖ from rat kidneys. Left: Urea-PAGE (lane 1: Low range ladder; lane 2: miRNA ladder; lanes 3-4: tRNAᴬˢᵖ from 6mo/36mo rats). Right: UV spectra. (H) tRNAᴬˢᵖ characterization. Top: TIC chromatograms (6mo vs 36mo). Bottom: Deconvoluted MS spectra of tRNA ᴬˢᵖ(GUC) and tRNA ᴬˢᵖ(manQUC) with sequence mapping. (I) UHPLC-QTOF-MS profile of RNase T1-digested tRNAᴬˢᵖ fragments. (J) Nucleoside analysis of purified tRNAᴬˢᵖ by UHPLC-QQQ-MS. (K) Senescence phenotypes in tRNA modification-deficient 2BS cells: SA- β -gal staining (left) and senescence marker expression (right). (L-O) Conservation of manQ deficiency across models: 2BS cells (L), D. melanogaster (M), paraquat-induced aging mice (N), (O) Human leukocyte (* p <0.05, ** p <0.01, *** p <0.001 vs aging/controls).

Article Snippet: Queuine hydrochloride (CAS: 69565-92-0, TRC-Q525000-0.5MG), was purchased from Toronto Research Chemicals.

Techniques: Modification, Control, Isolation, Biomarker Discovery, Tandem Mass Spectroscopy, Quantitation Assay, Purification, Sequencing, Staining, Marker, Expressing

Journal: bioRxiv

Article Title: Evolutionarily Conserved Decline of tRNA Mannosyl-Queuosine Links Translational Regulation to Aging and Is Reversed by Queuine

doi: 10.64898/2026.03.22.713446

Figure Lengend Snippet: Targeted manipulation of manQ tRNA modification regulates cellular senescence (A) Biosynthetic pathway of manQ tRNA modification in humans. (B) Age-dependent dysregulation of manQ pathway enzymes: Left: Relative expression analysis in human leukocytes (young vs aged donors). Right: qPCR validation in rat kidneys (6mo vs 36mo) (C-D) UHPLC-MS quantification of manQ modification: (C) 2BS cells cultured in queuine-supplemented horse serum. (D) Naturally senescent 2BS cells (* p <0.05, ** p <0.01). (E-G) Transcriptomic profiling of queuine-treated senescent 2BS cells: (E) Volcano plot of differentially expressed genes (|log₂FC|>2, FDR<0.05). (F) Top enriched KEGG pathways (bubble size: gene count; color: -log₁₀[p-value]). (G) GSEA showing JAK-STAT pathway suppression (NES=-2.1, FDR=0.002) (H) Human JAK-STAT signaling schematic highlighting queuine’s putative targeting. (I) Molecular docking of queuine in JAK2 kinase domain (binding affinity: −4.61 kcal/mol). (J) RT-qPCR showing JAK2 downregulation in queuine-treated 2BS cells. (K-L) MAN2C1 knockdown consequences: (K) manQ reduction by UHPLC-MS (** p <0.01). (L) Senescence marker induction (p16/p21). (M-N) MAN2C1 overexpression effects: (M) manQ restoration (** p <0.01 vs control). (N) Senescence marker suppression (** p <0.01).

Article Snippet: Queuine hydrochloride (CAS: 69565-92-0, TRC-Q525000-0.5MG), was purchased from Toronto Research Chemicals.

Techniques: Modification, Expressing, Biomarker Discovery, Cell Culture, Binding Assay, Quantitative RT-PCR, Knockdown, Marker, Over Expression, Control

Journal: bioRxiv

Article Title: Evolutionarily Conserved Decline of tRNA Mannosyl-Queuosine Links Translational Regulation to Aging and Is Reversed by Queuine

doi: 10.64898/2026.03.22.713446

Figure Lengend Snippet: Systemic queuine supplementation counteracts metabolic dysregulation in aging. (A-B) Plasma metabolomics profiling: (A) Proportional distribution of metabolite classes, (B) Shared/unique metabolites across groups (young/aged/aged+queuine). (C-D) Dysregulated metabolites in aged vs. queuine-treated mice: (C) Volcano plot (|log 2 FC|>2, FDR<0.05), (D) Hierarchical clustering of altered metabolites. (E-F) Pathway-specific metabolite restoration: (E) Prostaglandins, (F) Kynurenine pathway (* p <0.05,** p <0.01). (G) Integrative model: Queuine-mediated manQ restoration modulates 12 hallmarks of aging.

Article Snippet: Queuine hydrochloride (CAS: 69565-92-0, TRC-Q525000-0.5MG), was purchased from Toronto Research Chemicals.

Techniques: Clinical Proteomics

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Interaction analysis of the control drug Gemcitabine and the proposed nutraceutical therapeutics Queuine and Thiamine with the MAP4K4 binding site. ( A ) 3D binding pose of Gemcitabine (left) and 2D representation (right) of Gemcitabine docked in the MAP4K4 binding site. ( B ) 3D and 2D binding pose of Queuine ( C ) 3D and 2D binding pose of Thiamine. Coloring schemes: hydrogen bonding (purple arrows), pi-pi stacking (green line), and pi-cation interactions (red line)

Article Snippet: Queuine hydrochloride (Santa Cruz) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Control, Binding Assay

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Protein-ligand complex root mean square deviation (RMSD) ( A ) and root mean square fluctuation (RMSF) ( B ) plots for the apo, Gemcitabine, Queuine, and Thiamine bound forms of the MAP4K4 protein

Article Snippet: Queuine hydrochloride (Santa Cruz) was dissolved in dimethyl sulfoxide (DMSO).

Techniques:

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Schematic showing the durability of ligand-protein interactions during MD simulations. ( A ) Queuine-protein contacts prolonged for more than 50% of the simulation time. ( B ) Thiamine has shorter-lived less prolonged contacts

Article Snippet: Queuine hydrochloride (Santa Cruz) was dissolved in dimethyl sulfoxide (DMSO).

Techniques:

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Final snapshots at the end of the 100 ns MD simulations for Queuine ( A ), Thiamine ( B ), and Gemcitabine ( C )

Article Snippet: Queuine hydrochloride (Santa Cruz) was dissolved in dimethyl sulfoxide (DMSO).

Techniques:

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Viability of Panc-1 cells compared with that of the control group after exposure to different concentrations of Queuine ( A ), Thiamine ( B ) and Gemcitabine ( C ) for 48 h. The results represent the average ± SEM of the results from each experiment, which were repeated three times at different times with the same concentration ranges. The statistical significance was evaluated according to a p-value of < 0.05.Compared with the control, *p< \0.05, **p< \0.01

Article Snippet: Queuine hydrochloride (Santa Cruz) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Control, Concentration Assay

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Effects of different concentrations of Queuine and Gemcitabine on MAP4K4 activity. Absorbance values on the y-axis represent the enzyme amount measured using an ELISA kit, with different concentrations of Queuine ( A ) and Gemsitabine ( B ) indicated on the x-axis. The differences between the bar pairs marked with * ( p < 0.05) and *** ( p < 0.001) are significant

Article Snippet: Queuine hydrochloride (Santa Cruz) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Viability of Panc-1 cells compared with that of the control group after exposure to combined treatment of Queuine (QUE) and Gemcitabine (GEM) for 48 h. A ) Queuine (0.25 µM) with gemcitabine combination, B ) Queuine (1.25 µM) with gemcitabine combination

Article Snippet: Queuine hydrochloride (Santa Cruz) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Control

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Synergetic effects of Queuine and Gemcitabine on MAP4K4 activity. Absorbance values represent the enzyme amount measured using an ELISA kit. The difference between bar pairs marked with *** are significant at p < 0.001

Article Snippet: Queuine hydrochloride (Santa Cruz) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of the Turkish Chemical Society Section A: Chemistry

Article Title: Competitive Inhibition and Synergistic Effects of Nutraceutical and Metabolite Molecules on Anti-Acetylcholinesterase Activity

doi: 10.18596/jotcsa.1367877

Figure Lengend Snippet: Figure 2: Binding site interactions of lead compounds Thiamine (A), Queuine (B), and Etoperidone (C) with

Article Snippet: Etoperidone hydrochloride (Catalog No: sc-211494) and Queuine hydrochloride (Catalog No: sc-394021) were obtained from Santa Cruz Biotechnology.

Techniques: Binding Assay