Journal: bioRxiv

Article Title: Persistent interferon signaling that causes sensory neuron plasticity and pain in arthritis

doi: 10.1101/2025.01.18.633447

Figure Lengend Snippet: ( a ) Expression changes of Ifna and Ifnb mRNA levels at different times (hours and days) after autoantibody injection in mouse DRGs by qPCR. Pan- Ifna primers were used for all Ifna genes. Each time point of arthritis contains 4 mice, whereas control group contains 9 mice, * p < 0.05, ** p < 0.01, *** p < 0.001. ( b ) Representative RNAscope images of Ifna mRNA (top) and Ifnar1 mRNA (bottom) expression in control mouse DRGs. Scale bar = 25 µm. ( c ) Expression levels of type I IFN receptors Ifnar1 and Ifnar2 in different cell types from mouse DRG scRNA-seq dataset. ( d ) Illustration of transcriptional and translational pathways activated by type I interferons. ( e ) Currentclamp responses of cultured mouse DRG neurons to IFNα3 during different current ramp injections. ( f ) Plots of number of spikes recorded from control and IFNα3 stimulated DRG neurons under different ramp currents (control, n = 11; IFNα3, n = 9, * p < 0.05). ( g ) Phosphorylation of eIF4E (Ser209) (normalized to eIF4E) in mouse DRGs by western blotting at different times after antibody injection. Each time point contains 4 mice, except control group (before injection), which includes 6 mice, * p < 0.05, ** p < 0.01, *** p < 0.001. ( h ) Effect on phosphorylation of eIF4E by i.p. injection of IFNAR1 blocking antibody (IFN block) in arthritis mice (con, control mice, n = 4 in each group, * p < 0.05, ** p < 0.01). The fifth control sample labeled with an asterisk on GAPDH band was not included for quantification. ( i ) Representative RNAscope images of human DRGs from female healthy donors showing IFNAR1 mRNA ( TRPV1 / IFNAR1 , left) and IFNAR2 mRNA ( SOX10 / IFNAR2 , right) expression in neurons. Inset for IFNAR2 on the right showed co-localization of SOX10 and IFNAR2 also in satellite glia cells. Scale bar = 100 µm. ( j ) Western blot analysis shows IFNα expression in human DRGs from healthy controls (n = 11), RA patients with pain (Arthritis w pain, n = 3) and RA patients without pain (Arthritis wo pain, n = 5). ( k ) Quantification of IFNα expression by western blotting in ( j ) within healthy control, RA patients with pain (w pain) and RA patients without pain (wo pain), *** p < 0.001. ( l ) mRNA expression fold changes of IFNA, IFNB, IL1B, IL6 and TNF in human DRGs (* p < 0.05, *** p < 0.001). ( m ) mRNA expression fold changes of Ifna, Ifnb, Il1b and Il6 in mouse DRGs from control (n = 5), arthritis (n = 5) and arthritis mice treated with IFNAR1 blocking antibody (IFN block, n = 5), *p < 0.05, **p < 0.01. ( n ) Action potential responses in the current clamp setting following current injections of cultured mouse DRG neurons exposed to IFNα3 with or without MNK1/2 inhibitor eFT508. ( o ) Plots of number of spikes recorded from IFNα3 stimulated DRG neurons in presence or absence of MNK1/2 inhibitor under different ramp currents (IFNα3 + Vehicle, n = 14; IFNα3 + eFT508, n = 15, * p < 0.05). qPCR and western blotting data were analyzed with one-way ANOVA followed by Turkey’s multiple comparisons test, whereas patch clamp analysis was performed with two-way ANOVA followed by Šídák’s multiple comparisons test.

Article Snippet: A single i.p. injection of MNK1/2 inhibitor, Tomivosertib (eFT508/HY-100022, MCE) on day 48 after antibody injection in C57BL/6N mice (1 mg/kg, in DMSO:PEG300:Tween-80:Saline of 5:40:5:50), and then mechanical sensitivity of von Frey withdrawal threshold as well as nocifensive behavior of 2g von Frey and clip squeeze tests were measured around 1.5h and 24h after Tomivosertib administration; and on day 55 sunflower seed assay and inverted screen test were checked around 1.5h and 24h after Tomivosertib administration (n = 10, 5 females and 5 males). eIF4E/eIF4G interaction inhibitor, 4EGI-1 (324517, Sigma) was i.p. injected into antibody-induced arthritis C57BL/6N mice (15 mg/kg, in DMSO:PEG300:Tween-80:Saline of 5:40:5:50, on day 56).

Techniques: Expressing, Injection, Control, RNAscope, Cell Culture, Western Blot, Blocking Assay, Labeling, Patch Clamp

Journal: bioRxiv

Article Title: Persistent interferon signaling that causes sensory neuron plasticity and pain in arthritis

doi: 10.1101/2025.01.18.633447

Figure Lengend Snippet: ( a ) Systemic administration of IFNAR1 mAb (i.p., 40 mg/kg) reversed chronic joint pain, impairment of dexterity and limb function in the squeeze test, sunflower seed assay and inverted screen test (C57BL/6N, n = 5, ** p < 0.01). ( b ) Oral administration of TYK2 inhibitor (15 mg/kg) similarly reversed sensory deficits associated with arthritis (n = 8, * p < 0.05, ** p < 0.01). ( c ) MNK1/2 inhibitor (i.p., eFT508, 1 mg/kg) reversed sensory deficits associated with arthritis (n = 10, * p < 0.05, ** p < 0.01). ( d ) eIF4E inhibitor (i.p., 4EGI, 15 mg/kg) reversed sensory deficits associated with arthritis (n = 6, * p < 0.05, ** p < 0.01). In all graphs show behavior prior to inducing arthritis (BL), at the post-inflammatory phase following antibody-induced arthritis (Arthritis), post inflammatory phase of arthritis 1.5h after administration of compounds (1.5hr) and after washout of compounds (24hr or 48hr). Squeeze test and sunflower seed assay were analyzed with Fridman test followed by Dunn’s multiple comparisons test. The inverted screen test was performed with one-way ANOVA followed by Šídák’s multiple comparisons test.

Article Snippet: A single i.p. injection of MNK1/2 inhibitor, Tomivosertib (eFT508/HY-100022, MCE) on day 48 after antibody injection in C57BL/6N mice (1 mg/kg, in DMSO:PEG300:Tween-80:Saline of 5:40:5:50), and then mechanical sensitivity of von Frey withdrawal threshold as well as nocifensive behavior of 2g von Frey and clip squeeze tests were measured around 1.5h and 24h after Tomivosertib administration; and on day 55 sunflower seed assay and inverted screen test were checked around 1.5h and 24h after Tomivosertib administration (n = 10, 5 females and 5 males). eIF4E/eIF4G interaction inhibitor, 4EGI-1 (324517, Sigma) was i.p. injected into antibody-induced arthritis C57BL/6N mice (15 mg/kg, in DMSO:PEG300:Tween-80:Saline of 5:40:5:50, on day 56).

Techniques:

Journal: Frontiers in Immunology

Article Title: MicroRNA-1985 enhances the redox capability of scallop ( Patinopecten yessoensis ) in response to poly(I:C) stimulation by targeting MNK1

doi: 10.3389/fimmu.2025.1556591

Figure Lengend Snippet: Nucleotide and predicted amino acid sequences of PyMNK1 . The numbers on the left alternate from top to bottom as the nucleotide sequence and the deduced amino acid sequence numbers; the black font indicates the nucleotide sequence, and the red font indicates the amino acid sequence. The domain is shaded in green. The start codon (ATG) is framed by a blue box; the asterisk (*) represents the termination codon (TGA), and the polyadenylation signal (AATAA) is framed by a box.

Article Snippet: Specific small interfering RNAs (siRNAs) targeting PyMNK1 (si MNK1 ) and RNA interference negative controls (NC-RNAi) were designed and synthesized by Sangon Biotech (Shanghai, China; ).

Techniques: Sequencing

Journal: Frontiers in Immunology

Article Title: MicroRNA-1985 enhances the redox capability of scallop ( Patinopecten yessoensis ) in response to poly(I:C) stimulation by targeting MNK1

doi: 10.3389/fimmu.2025.1556591

Figure Lengend Snippet: Sequence and CpG island predictions of the SOD promotor, binding site identification, and verification of the PyMNK1 and SOD promotor. (A) The sequence of the SOD promoter. The transcription initiation site (TSS) is in the red font. The blue box represents the start codon (ATG). (B) CpG island prediction of the SOD promotor from Patinopecten yessoensis and five other species. (C) Schematic representation of putative Py MNK1 targeting sites and regulatory elements in the SOD promotor. (D) An analysis of relative luciferase activity. Different lower-case letters (e.g., a, b) represent the significance levels between different groups ( P < 0.05). ** represent P < 0.01 vs . SOD WT1 binding sites group. ## represent P < 0.01 vs . SOD WT2 binding sites group. pGL3-basic, promoter vector; pGL3-control, positive control; WT, wild type.

Article Snippet: Specific small interfering RNAs (siRNAs) targeting PyMNK1 (si MNK1 ) and RNA interference negative controls (NC-RNAi) were designed and synthesized by Sangon Biotech (Shanghai, China; ).

Techniques: Sequencing, Binding Assay, Luciferase, Activity Assay, Plasmid Preparation, Control, Positive Control

Journal: Frontiers in Immunology

Article Title: MicroRNA-1985 enhances the redox capability of scallop ( Patinopecten yessoensis ) in response to poly(I:C) stimulation by targeting MNK1

doi: 10.3389/fimmu.2025.1556591

Figure Lengend Snippet: Spatial expression pattern, binding site identification, and verification of miR-1985 and PyMNK1 . (A) Spatial expression pattern of miR1985 and PyMNK1. mRNA. HP: hepatopancreas; GO: gonads; MU: muscles; GI: gills; MA: mantles; HE: hemolymph. (B) Spatial expression pattern of Py MNK1 protein. (C, D) Schematic representation of the putative miR-1985 targeting site in PyMNK1 mRNA and an analysis of relative luciferase activity. Different lower-case letters (e.g., a or a’) indicate significant differences between different tissues ( P < 0.05). “**” represents an extremely significant difference between miR-1985 and Py MNK1 mRNA in the same tissue or between NC + PyMNK1 -WT and miR-1985 + PyMNK1 -WT ( P < 0.01).

Article Snippet: Specific small interfering RNAs (siRNAs) targeting PyMNK1 (si MNK1 ) and RNA interference negative controls (NC-RNAi) were designed and synthesized by Sangon Biotech (Shanghai, China; ).

Techniques: Expressing, Binding Assay, Muscles, Luciferase, Activity Assay

Journal: Frontiers in Immunology

Article Title: MicroRNA-1985 enhances the redox capability of scallop ( Patinopecten yessoensis ) in response to poly(I:C) stimulation by targeting MNK1

doi: 10.3389/fimmu.2025.1556591

Figure Lengend Snippet: Effects of PyMNK1 and miR-1985 on mRNA expression, ROS content, and total SOD and CAT enzymatic activity changes in gills of Patinopecten yessoensis . (A, B) Effects of PyMNK1 and miR-1985 on PyMNK1 mRNA and protein expression. (C) Effects of PyMNK1 and miR-1985 on ROS content. (D-F) Effects of PyMNK1 and miR-1985 on SOD mRNA expression and enzymatic activity of SOD and CAT. I: negative control of PyMNK1 silencing group (NC-RNAi); II: PyMNK1 silencing group (RNAi); III: negative control of miR-1985 overexpression group (NC-Agomir); IV: miR-1985 overexpression group (Agomir); V: negative control of miR-1985 inhibition group (NC-Antagomir); VI: miR-1985 inhibition group (Antagomir). * represents P < 0.05; ** represents P < 0.01.

Article Snippet: Specific small interfering RNAs (siRNAs) targeting PyMNK1 (si MNK1 ) and RNA interference negative controls (NC-RNAi) were designed and synthesized by Sangon Biotech (Shanghai, China; ).

Techniques: Expressing, Activity Assay, Negative Control, Over Expression, Inhibition

Journal: Frontiers in Immunology

Article Title: MicroRNA-1985 enhances the redox capability of scallop ( Patinopecten yessoensis ) in response to poly(I:C) stimulation by targeting MNK1

doi: 10.3389/fimmu.2025.1556591

Figure Lengend Snippet: Poly(I:C) stimulation altered miR-1985, Py MNK1 (mRNA and protein), and SOD mRNA expression, ROS content, and total SOD and CAT enzymatic activity in gills of Patinopecten yessoensis . (A-C) Relative expression of miR-1985 and Py MNK1 (mRNA and protein) in gills of P. yessoensis after poly(I:C) stimulation. (D–F) Changes in the ROS content and total SOD and CAT enzymatic activity in gills of P. yessoensis after poly(I:C) stimulation. (G) Relative expression of SOD mRNA in gills of P. yessoensis after poly(I:C) stimulation. (H) Relative expression of miR-1985, PyMNK1 mRNA, and SOD mRNA after poly(I:C) stimulation. * represents P < 0.05 vs. 0 hps; ** represent P < 0.01 vs. 0 hps.

Article Snippet: Specific small interfering RNAs (siRNAs) targeting PyMNK1 (si MNK1 ) and RNA interference negative controls (NC-RNAi) were designed and synthesized by Sangon Biotech (Shanghai, China; ).

Techniques: Expressing, Activity Assay

Journal: Frontiers in Immunology

Article Title: MicroRNA-1985 enhances the redox capability of scallop ( Patinopecten yessoensis ) in response to poly(I:C) stimulation by targeting MNK1

doi: 10.3389/fimmu.2025.1556591

Figure Lengend Snippet: Effects of miR-1985 expression and PyMNK1 silencing on PyMNK1 (mRNA and protein) expression, ROS content, and total enzymatic activity of SOD and CAT in gills of Patinopecten yessoensis after poly(I:C) stimulation. (A–C) Effects of miR-1985 expression and PyMNK1 silencing on ROS content, total SOD and CAT enzyme activity in gills of P. yessoensis after poly(I:C) stimulation. (D–E) Effects of miR-1985 expression and PyMNK1 silencing on Py MNK1 (mRNA and protein) expression in gills of P. yessoensis after poly(I:C) stimulation. (F) Effects of miR-1985 expression and PyMNK1 silencing on SOD mRNA expression in gills of P. yessoensis after poly(I:C) stimulation. (G) Schematic diagrams of miR-1985 and PyMNK1 involved in responding to poly(I:C) stimulation in P. yessoensis . Schematic images were drawn by Figdraw ( http://www.figdraw.com ). NC-RNAi, negative control of PyMNK1 silencing group; RNAi, PyMNK1 silencing group; NC-Agomir, negative control of miR-1985 overexpression group; Agomir, miR-1985 overexpression group; NC-Antagomir, negative control of miR-1985 inhibition group; Antagomir, miR-1985 inhibition group. * represents P < 0.05; ** represents P < 0.01.

Article Snippet: Specific small interfering RNAs (siRNAs) targeting PyMNK1 (si MNK1 ) and RNA interference negative controls (NC-RNAi) were designed and synthesized by Sangon Biotech (Shanghai, China; ).

Techniques: Expressing, Activity Assay, Negative Control, Over Expression, Inhibition