Long Non-Coding RNAs Associated with Heterochromatin Function in Immune Cells in Psychosis
<p><b>Recruitment of PRC2 complex on the heterochromatin assembly.</b> Assembling the PRC2 components on sequence specific DNA mediated by lncRNAs (MEG3, PINT and GAS5). The PRC2 complex is shown with its core components (EZH2, SUZ12, EED and RBBP4/7) and this is responsible for di- and tri-methylation of nucleosomal histone proteins (H3K27me3). JARID2, a regulatory component of PRC2, interacts with the lncRNA MEG3 (blue dashed line) and helps the recruitment of the PRC2 to the chromatin. The lncRNAs (PINT or GAS5 denoted by the brown dotted line) cobble around the core component of PRC2 and help in its recruitment to the chromatin.</p> "> Figure 2
<p><b>Diagnostic difference in the expression of lncRNAs.</b> A difference in the expression of lncRNAs MEG3, PINT and GAS-5 was seen between control participants and participants with psychosis. Participants with psychosis had higher levels of MEG3, but lower levels of PINT and GAS5. Statistical significance is determined by <span class="html-italic">t</span>-test (* <span class="html-italic">p</span> < 0.05; *** <span class="html-italic">p</span> < 0.001). The figures represent Mean ± SEM for the relative expression of lncRNAs. SEM: Standard Error of Mean.</p> "> Figure 3
<p><b>LncRNA expression by illness acuity status.</b> Entire participant sample was used to determine the expression of the lncRNAs relative to illness acuity status. There were significant differences found in regard to inpatient or outpatient status. Statistical significance is determined using ANOVA (* <span class="html-italic">p</span> < 0.05; *** <span class="html-italic">p</span> < 0.001; ns, not significant). The figures represent Mean ± SEM for the relative expression of lncRNAs. SEM: Standard Error of Mean.</p> "> Figure 4
<p><b>LncRNA expression between controls and participants on antipsychotic medication</b>. Yes = Participants with the diagnosis of psychosis and currently on antipsychotic medication, No = Control participants who are not on antipsychotic medication. MEG3 is significantly higher, and PINT and GAS5 lower in those taking antipsychotic medication when compared to those not currently on antipsychotics. Statistical significance is determined by <span class="html-italic">t</span>-test (* <span class="html-italic">p</span> < 0.05; ** <span class="html-italic">p</span> < 0.01; *** <span class="html-italic">p</span> < 0.001; ns, not significant). The figures represent Mean ± SEM for the relative expression of lncRNAs. SEM: Standard Error of Mean.</p> "> Figure 5
<p><b>LncRNA expression between drug naïve psychotic participants and participants on antipsychotic medication.</b> Antipsychotic medication = Participants with the diagnosis of psychosis and on antipsychotic medication. Drug naïve = Participants with a diagnosis of psychosis who are not on antipsychotic medication. MEG3 expression was significantly downregulated in participants currently on antipsychotic medication when compared to drug naïve psychotic participants. There were no differences in PINT and GAS5 expression levels between groups. Statistical significance is determined by one-way ANOVA (* <span class="html-italic">p</span> < 0.05; ns, not significant). The figures represent Mean ± SEM for the relative expression of lncRNAs. SEM: Standard Error of Mean.</p> "> Figure 6
<p><b>LncRNA expression between participants with risperidone treatment and participants on other antipsychotic medications.</b> Risperidone = participants with the diagnosis of psychosis and on risperidone treatment alone. Other antipsychotics = participants with the diagnosis of psychosis who are not on risperidone medication, but on other antipsychotic medication. This group does not include drug naïve participants. Statistical significance is determined by one-way ANOVA and Tukey’s post hoc test (*** <span class="html-italic">p</span> < 0.001; ns, not significant). The figures represent Mean ± SEM for the relative expression of lncRNAs. SEM: Standard Error of Mean.</p> "> Figure 7
<p><b>Comparison of lncRNA MEG3 expression levels from four treatment conditions</b>: controls, risperidone only, currently on any other antipsychotic, and drug naïve. Statistical significance is determined by <span class="html-italic">t</span>-test (** <span class="html-italic">p</span> < 0.01; *** <span class="html-italic">p</span> < 0.001; ns, not significant). The figures represent Mean ± SEM for the relative expression of lncRNAs. SEM: Standard Error of Mean.</p> "> Figure 8
<p>Antipsychotic response on induced M2<sup>tol</sup> macrophages. MEG3 was significantly upregulated in response to LPS treatment, an effect compounded by Risperidone co-treatment, a pattern also seen with GAS-5. PINT was upregulated in response to LPS, but this effect was ablated with Risperidone treatment. The experiment was conducted in duplicate (<span class="html-italic">n</span> = 2), and M2<sup>tol</sup> levels are compared to M0 set arbitrarily as the baseline. * <span class="html-italic">p</span> < 0.05; ** <span class="html-italic">p</span> < 0.01; *** <span class="html-italic">p</span> < 0.001; ns, not significant, as determined by ANOVA (<span class="html-italic">n</span> = 2).</p> "> Figure 9
<p><b>Sample selection for analysis of lncRNA expression</b>. This diagram shows the total sample of study participants. First the total population was grouped by diagnosis. Then the psychosis group was further split by participants who received antipsychotic treatment and those participants who were drug naïve. The antipsychotic treated group was further sorted into those who received risperidone alone and participants who received all other antipsychotics except risperidone.</p> ">
Abstract
:1. Introduction
2. Results
2.1. Demographics
2.2. Diagnostic Differences
2.3. Co-Expression of lncRNAs
2.4. Differences in the Expression of lncRNAs Based on Antipsychotic Treatment Status in the Clinical Sample
2.5. Effect of Antipsychotics on the expression of lncRNAs in the M0 and M2 macrophages
3. Discussion
4. Materials and Methods
4.1. Participant Information and Clinical Measures
4.2. Sample Collection, Processing and Quantitative real-time PCR (qRT-PCR)
4.3. Primary Clinical Measures
4.4. THP1-Derived M2tol Polarized Macrophages and Risperidone Treatment
4.5. Statistical Analyses
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Control | Psychosis | Group Differences | ||||
---|---|---|---|---|---|---|
Total (n) | 44 (33.8%) | 86 (66.2%) | ||||
Sex | Female (n) | 25 (41.0%) | 36 (59.0%) | χ2(3) = 2.615, p = 0.106 | ||
Male (n) | 19 (27.5%) | 50 (72.5%) | ||||
Race | Caucasian, non-Hispanic (n) | 12 (54.5%) | 10 (45.5%) | χ2(3) = 22.494, p < 0.001 | ||
Black, non-Hispanic (n) | 18 (22.5%) | 62 (77.5%) | ||||
Asian or other Pacific Islander (n) | 10 (83.3%) | 2 (16.7%) | ||||
Hispanic (n) | 4 (25.0%) | 12 (75.0%) | ||||
Antipsychotic Use | Yes (n) | 0 | 77 | |||
No (n) | 44 | 9 | ||||
Other medication * | Yes (n) | 4 | 9 | |||
No (n) | 40 | 77 | ||||
Smoking | Yes(n) | 7 | 39 | χ2(3) = 11.034, p = 0.001 | ||
No (n) | 37 | 47 | ||||
Age (Mean ± SD) | 38.77 ± 12.82 | 40.21 ± 12.98 | t(128) = −0.600, p = 0.55 | |||
BMI (Mean ± SD) | 28.93 ± 8.1 | 30.8 ± 8.15 | t(128) = 1.241, p = 0.217 | |||
Mean | SD | Mean | SD | |||
Age at onset of illness | ~ | ~ | 21.32 | 7.88 | ||
Duration of untreated psychosis | ~ | ~ | 2.97 | 5.85 | ||
Duration of illness | ~ | ~ | 19.33 | 14.35 |
N = 130 | Control (n = 44) | Psychosis (n = 86) | Group Differences | |||||
---|---|---|---|---|---|---|---|---|
Inpatient(n = 33) | Outpatient(n = 53) | |||||||
PANSS Five-Factor Scores | Mean ± SD | Range | Mean ± SD | Range | Mean ± SD | Range | Inpatient and Outpatient † | Control, Inpatient and Outpatient ‡ |
Positive | 4.11 ± 0.49 | 4–7 | 18.27 ± 2.39 | 13–23 | 13.49 ± 4.40 | 5–21 | t(84) = 5.73 *** | F(2,127) = 218.20 *** |
Negative | 6.55 ± 7.56 | 6–15 | 20.76 ± 7.56 | 6–36 | 15.60 ± 5.91 | 6–27 | t(84) = 3.58 *** | F(2,127) = 68.56 *** |
Cognitive | 5.48 ± 1.07 | 5–10 | 17.34 ± 4.09 | 8–25 | 12.00 ± 4.01 | 5–21 | t(84) = 5.95 *** | F(2,127) = 121.25 *** |
Excitement | 5.66 ± 1.98 | 4–13 | 10.91 ± 3.41 | 6–28 | 9.34 ± 2.78 | 4–16 | t(84) = 2.33 * | F(2,127) = 39.35 *** |
Depression | 7.55 ± 2.71 | 5–14 | 15.69 ± 2.80 | 10–23 | 13.83 ± 3.33 | 6–22 | t(84) = 2.68 ** | F(2,127) = 83.173 *** |
PINT | GAS5 | |
---|---|---|
MEG3 | * r = 0.237 p = 0.015 | * r = 0.080 p = 0.422 |
PINT | * r = 0.222 p = 0.016 |
GAPDH | 5′-CGAGATCCCTCCAAAATCAA-3′ | 5′-TTCACACCCATGACGAACAT-3′ |
ACTB | 5′s-TGAAGGTAGTTTCGTGGATGC-3′ | 5′-TCCCTGGAGAAGAGCTACGA-3′ |
MEG3 | 5′-GCGGAGAGCAGAGAGGGA-3′ | 5′-AGGAGAGACCCGGGTGAG-3′ |
PINT | 5′-CCATCTGGAGTTTCTCTGCCT-3′ | 5′-GGTAAGACTCTGTCTTCAGCTGTTA-3′ |
GAS-5 | 5′-TCCTGTGAGGTATGGTGCTGG-3′ | 5-AACTTGCCTGGACCAGCTTA-3′ |
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Sudhalkar, N.; Rosen, C.; Melbourne, J.K.; Park, M.R.; Chase, K.A.; Sharma, R.P. Long Non-Coding RNAs Associated with Heterochromatin Function in Immune Cells in Psychosis. Non-Coding RNA 2018, 4, 43. https://doi.org/10.3390/ncrna4040043
Sudhalkar N, Rosen C, Melbourne JK, Park MR, Chase KA, Sharma RP. Long Non-Coding RNAs Associated with Heterochromatin Function in Immune Cells in Psychosis. Non-Coding RNA. 2018; 4(4):43. https://doi.org/10.3390/ncrna4040043
Chicago/Turabian StyleSudhalkar, Niyati, Cherise Rosen, Jennifer K. Melbourne, Mi Rae Park, Kayla A. Chase, and Rajiv P. Sharma. 2018. "Long Non-Coding RNAs Associated with Heterochromatin Function in Immune Cells in Psychosis" Non-Coding RNA 4, no. 4: 43. https://doi.org/10.3390/ncrna4040043