selective prmt4 inhibitors  (MethylGene Inc)

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: A flowchart summarises groups of animals used in different experiments and the schedule of nose-poke operant conditioning. The animals were habituated to the operant chamber (Days 1 to 4) and divided into 5 experiments and 13 groups. Experiment 1: non-conditioned animals (Group 1) were subjected to an operant chamber, but no pellets were dispensed. The conditioned animals (Group 2) were presented with sucrose pellet in response to poking at the active port. The Group 2 rats were trained for sucrose pellet self-administration for 15 min twice/day from the 5th to the 11th day. On the 12th day, one group of rats was subjected to OFT followed by a probe trial, whilst the other group was subjected to LDB followed by a probe trial. These rats were killed 15 min after the probe trial, and one cohort was used for molecular biology, whereas the other was perfused for immunofluorescence analysis. Experiments 2–4: after habituation, animals were operated for cannula implantation and were allowed to recover (5th to 11th day). The cannulated animals were subjected to operant conditioning for 15 min/day twice a day till a steady baseline was achieved (12th to 18th). Experiment 2: the PRMT4 siRNA (Groups 4, 5) and scrambled siRNA (Group 3) were infused in the basolateral amygdala of the conditioned animals on the 18th day after the training session. Experiment 3: PRMT4 (CARM1) inhibitor and aCSF were administered on the 18th day after the training session in rats belonging to Groups 7, 8 and 6, respectively. Animals belonging to Groups 3, 4, 6 and 7 were tested (OFT, LDB and probe trial) on the 19th day (24 h post-infusion) and were killed 15 min after the probe trial. Group 5 and 8 rats were tested for nose poking from the 19th to the 22nd day. On the 23rd day,OFT, LDB and probe trials were conducted, and rats were sacrificed 15 min after the probe trial. Experiment 4: the PRMT4 siRNA/PRMT4 inhibitor/aCSF-treated animals belonging to Groups 9–11 were subsequently treated with NPY peptide on the 19th day, and OFT, LDB, followed by probe trial, were conducted 1 h after NPY infusion. Brains were isolated for the molecular analyses immediately after the probe trial in Groups 1–11. *Separate groups of rats were generated for immunohistochemistry. Experiment 5: cannulated animals were divided into two groups before training. Rats belonging to Group 12 were infused with scrambled siRNA, whereas Group 13 was administered with PRMT4 siRNA on the 11th day and sacrificed on the 12th day

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Immunofluorescence, Isolation, Generated, Immunohistochemistry

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: Reward conditioning alters the expression of PRMTs and NPY in the amygdala. A Nose-poke activity of the rats trained for sucrose pellet self-administration and non-conditioned controls for 6 days (from Days 6 to 11 in Experiment 1). B Nose-poke activity of the rats in active port during 5 min probe trial (* p < 0.05; ** p < 0.01; *** p < 0.001 versus non-conditioned). C Schematic representation of the amygdala region used for the analysis. Effect of conditioning on the D mRNA levels of different PRMTs, E , F protein levels of PRMT4 and c-FOS in the amygdala. Graph representing G mRNA and H protein expression of NPY in the conditioned and non-conditioned rats (** p < 0.01; *** p < 0.001 versus non-conditioned). Values ( n = 5/group) are represented as means (± SEM)

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Expressing, Activity Assay

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: PRMT4 and CBP-mediated histone modifications are enriched at the NPY promoter of conditioned rats. A Schematic representation of the NPY promoter. Effect of reward conditioning on B PRMT4 , C H3R17me2a levels at NPY promoter. D Co-immunoprecipitation (Co-IP) and reverse immunoprecipitation assays showing interactions of PRMT4 with CBP. Effect of reward conditioning on E CBP occupancy, F H3K14ac and G PRMT4-CBP co-occupancy at NPY promoter (* p < 0.05, ** p < 0.01, *** p < 0.001 versus non-conditioned). Values ( n = 4–5/group) are represented as means (± SEM)

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: Reward conditioning enhances the expression of PRMT4 and NPY in the neurons of the basolateral amygdala (BLA). A Representative confocal images (20 × ) show PRMT4 (green) and NPY-positive cells (red), DAPI (blue), and their colocalization. The merged panels indicate NPY-positive cells co-expressing PRMT4 (arrows). PRMT4 predominantly colocalised with DAPI. The graph shows C PRMT4 immunoreactive area, and D the percentage of NPY-positive area in the BLA of conditioned and non-conditioned rats. The percentage of PRMT4-positive cells increased in the BLA of conditioned rats as compared to non-conditioned controls. Values ( n = 5/group) are represented as means (± SEM) and *** p < 0.001 versus non-conditioned

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Expressing

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: PRMT4-NPY axis in the basolateral amygdala is essential for the nose-poke activity of conditioned rats. A Schematic representation (left) and photomicrograph (right) of the coronal section of rat brain showing the position of the cannula targeted towards Basolateral Amygdala (BLA) at coordinates AP: − 3.14 mm, ML: ± 5 mm, DV: 4 mm (Paxinos and Watson ). Effect of intra-BLA infusion of B PRMT4 siRNA, C PRMT4 inhibitor on nose-poke activity of the reward conditioned rats (*** p < 0.001 versus non-conditioned; ### p < 0.001 versus conditioned + scrambled siRNA-24 h or conditioned + aCSF-24 h; $$$ p < 0.001 versus conditioned + PRMT4 siRNA or conditioned + PRMT4 inhibitor), D effect of NPY administration on PRMT4 siRNA/inhibitor-treated rats ( $$$ p < 0.001 versus conditioned + PRMT4 siRNA-24 h or conditioned + PRMT4 inhibitor-24 h, the number of nose pokes shown in the figure was recorded on the 19th day, 1 h after NPY administration during a 5-min probe trial session). Graphs representing the effect of intra-BLA infusion of scrambled siRNA, aCSF, PRMT4 siRNA, PRMT4 inhibitor, NPY peptide, and PRMT4 siRNA + NPY and PRMT4 inhibitor + NPY on E anxiety-like behaviours; L time spent in light compartment and D time spent in dark compartment, and F locomotion of conditioned rats (*** p < 0.001 versus conditioned + scrambled siRNA-24 h or conditioned + aCSF-24 h). Values ( n = 5–8/group) are represented as means (± SEM)

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Activity Assay

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: NPY expression in the amygdala of conditioned rats declined with the reduction in PRMT4 function by siRNA or inhibitor. Effect of A PRMT4 siRNA, B PRMT4 inhibitor and C PRMT4 siRNA/inhibitor co-administered with NPY peptide on PRMT4 mRNA expression in the amygdala. D Representative photomicrograph of western blot and E relative quantification of PRMT4 protein levels in conditioned rats infused with scrambled siRNA and PRMT4 siRNA. Graph representing the effect of F PRMT4 siRNA, G PRMT4 inhibitor and H PRMT4 siRNA/inhibitor co-administered with NPY peptide on NPY mRNA levels in the amygdala. I Graph representing NPY protein levels in PRMT4 siRNA-treated animals as estimated by ELISA. The intra-BLA infusion of PRMT4 siRNA/PRMT4 inhibitor caused a reduction in the PRMT4 and NPY expression, which was recovered in 5 days (** p < 0.01, *** p < 0.001 versus non-conditioned; ## p < 0.01, ### p < 0.001 versus Conditioned + aCSF/scrambled siRNA-24 h; $$$ p < 0.001 versus Conditioned + NPYp)

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Expressing, Western Blot, Quantitative Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: Effect of PRMT4 siRNA and PRMT4 inhibitor on the PRMT4 immunoreactivity in the BLA. Representative images (× 20) with PRMT4 (green) and NeuN-positive cells (red), and their colocalization (yellow). The merged panels indicate NeuN-positive cells co-expressing PRMT4 (arrows). PRMT4 predominantly colocalised with NeuN (C) Graph represents the intensity of PRMT4 immunofluorescence in different treatment groups (** p < 0.01 versus non-conditioned; ### p < 0.001 versus conditioned + scrambled siRNA-24 h)

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Expressing, Immunofluorescence

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: PRMT4-mediated histone arginine methylation regulates the NPY promoter. Effect of intra-BLA administration of scrambled siRNA, PRMT4 siRNA, NPY peptide and PRMT4 siRNA + NPY peptide on levels of A PRMT4, B H3R17me2a and C CBP occupancy at the NPY promoters. Values ( n = 5/group) are represented as means (± SEM) and (* p < 0.05, *** p < 0.001 versus non-conditioned; ### p < 0.001 versus conditioned + scrambled siRNA-24 h)

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Methylation

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: Effect of intra-BLA administration of PRMT4 siRNA in the naïve control rats on the levels of NPY mRNA ( A ), PRMT4 mRNA ( B ) and protein ( C and D ), and H3R17me2a ( E ) and PRMT4 ( F ) occupancy at the NPY promoter. Naïve rats infused with PRMT4 siRNA demonstrated a reduction in levels of PRMT4 and H3R17me2a at the NPY promoter, resulting in low NPY expression

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Control, Expressing

Journal: Cellular and Molecular Neurobiology

Article Title: Histone Arginine Methylation Regulates Neuropeptide Y Expression in the Basolateral Amygdala to Promote Reward-Seeking Behaviour

doi: 10.1007/s10571-025-01614-5

Figure Lengend Snippet: Schematic representation of NPY gene regulation in the amygdala and its effect on reward and reinforcement. Reward conditioning upregulates PRMT4, which in turn methylates 17th arginine in histone 3 (H3R17me2a) at the NPY promoter. PRMT4 cooperates with CBP and augments histone acetylation (H3K14ac) at the NPY promoter. These events remodel the NPY promoter to facilitate the NPY expression and promote reward-seeking activity. The PRMT4 siRNA infusion reversed the NPY promoter remodelling due to reward conditioning, attenuated NPY levels, and reduced nose-poke activity. NPY peptide administration following the siRNA infusion normalised the positive reinforcement. Therefore, PRMT4 in association with CBP seems to play an essential role in NPY expression within the framework of basolateral amygdala and reward processing

Article Snippet: The stock solution for PRMT4 inhibitor (217531, Merck) was prepared in DMSO to make 1.8 mM and was further diluted in aCSF to 100 μM and infused at the dosage of 25 μM/0.5 μl (IC 50 = 50.7 μM, Supplementary Fig. S2).

Techniques: Expressing, Activity Assay

Journal: Journal of cellular physiology

Article Title: Protein arginine methyltransferase 4 modulates nitric oxide synthase uncoupling and cerebral blood flow in Alzheimer's disease.

doi: 10.1002/jcp.30858

Figure Lengend Snippet: FIGURE 1 Aged female 3xTg‐AD mice exhibit differential expression of PRMT4 which can be reversed by TP‐064. Relative mRNA levels and protein of PRMTs in the cortex and hippocampus were measured by quantitative RT‐PCR and capillary‐based immunoassay, respectively. (a) Relative mRNA levels of PRMTs 1−9 were measured in brain tissue from aged, female C57 and 3xTg mice. (b, c) Relative protein expression of protein arginine methyltransferase 6 (PRMT6) (~45 kDA) (b) and PRMT4 (~63 kDA) (c) were measured via ProteinSimple capillary‐based immunoassay in cortical and hippocampal lysates from aged, female C57 and 3xTg animals. (d) Computer generated pseudo‐blot images depcting representative bands for PRMT4 and PRMT6. (e) PRMT4 inhibitorTP‐064 reduced PRMT4 protein expression in a dose‐dependent manner. Results are expressed as mean ± SEM, n indicates number of animals used. *p < 0.05, **p < 0.02, ***p < 0.01, v. control or untreated 3xTg, evaluated by one‐way ANOVA with Bonferroni (1a)/Tukey's (1e) post‐hoc analysis respectively and Student's t‐test as appropriate, (n = 3−6). PRMT4, protein arginine methyltransferase 4.

Article Snippet: Treatment consisted of specific PRMT4 inhibitor TP‐064 (Cat. No. 6008; Tocris) (Zhang et al., 2021; Zhong et al., 2018).

Techniques: Quantitative Proteomics, Quantitative RT-PCR, Expressing, Generated, Control

Journal: Journal of cellular physiology

Article Title: Protein arginine methyltransferase 4 modulates nitric oxide synthase uncoupling and cerebral blood flow in Alzheimer's disease.

doi: 10.1002/jcp.30858

Figure Lengend Snippet: FIGURE 5 PRMT4 inhibition via TP‐064 enhances regional cortical cerebral blood flow. Representative flux image of cortical vasculature via laser speckle contrast imaging (a). Aged 3xTg‐AD female mice had impaired cortical regional cerebral blood flow as compared to age/sex‐ matched control C57 mice, while treatment with TP‐064 (30 mg/kg, 7 days, IP) enhanced cerebral blood flow (b). Results are expressed as mean ± SEM, n indicates number of animals used, *p < 0.05 v. C57, #p < 0.05 v. 3xTg and C57 ± TP‐064 evaluated by two‐way ANOVA with Tukey's post hoc analyses. PRMT4, protein arginine methyltransferase 4.

Article Snippet: Treatment consisted of specific PRMT4 inhibitor TP‐064 (Cat. No. 6008; Tocris) (Zhang et al., 2021; Zhong et al., 2018).

Techniques: Inhibition, Imaging, Control

Journal:

Article Title: Effects of a Novel Arginine Methyltransferase Inhibitor on T Helper Cell Cytokine Production

doi: 10.1111/j.1742-4658.2010.07623.x

Figure Lengend Snippet: Table 1

Article Snippet: Recently, both Methylgene and Bristol-Myers Squibb have reported high potency (picomolar IC 50 ) and selective PRMT4 inhibitors, though the Methylgene compound was not active in cellular assays and no cellular data was reported for the Bristol-Myers Squibb compounds [ 28 – 30 ].

Techniques: Methylation

Journal:

Article Title: Effects of a Novel Arginine Methyltransferase Inhibitor on T Helper Cell Cytokine Production

doi: 10.1111/j.1742-4658.2010.07623.x

Figure Lengend Snippet: Table 1

Article Snippet: Recently, both Methylgene and Bristol-Myers Squibb have reported high potency (picomolar IC 50 ) and selective PRMT4 inhibitors, though the Methylgene compound was not active in cellular assays and no cellular data was reported for the Bristol-Myers Squibb compounds [ 28 – 30 ].

Techniques: Methylation