Data_Sheet_1_Deciphering Risperidone-Induced Lipogenesis by Network Pharmacology and Molecular Validation.PDF

BackgroundRisperidone is an atypical antipsychotic that can cause substantial weight gain. The pharmacological targets and molecular mechanisms related to risperidone-induced lipogenesis (RIL) remain to be elucidated. Therefore, network pharmacology and further experimental validation were undertaken to explore the action mechanisms of RIL.MethodsRILs were systematically analyzed by integrating multiple databases through integrated network pharmacology, transcriptomics, molecular docking, and molecular experiment analysis. The potential signaling pathways for RIL were identified and experimentally validated using gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) analysis.ResultsRisperidone promotes adipocyte differentiation and lipid accumulation through Oil Red O staining and reverse transcription-polymerase chain reaction (RT-PCR). After network pharmacology and GO analysis, risperidone was found to influence cellular metabolism. In addition, risperidone influences adipocyte metabolism, differentiation, and lipid accumulation-related functions through transcriptome analysis. Intersecting analysis, molecular docking, and pathway validation analysis showed that risperidone influences the adipocytokine signaling pathway by targeting MAPK14 (mitogen-activated protein kinase 14), MAPK8 (mitogen-activated protein kinase 8), and RXRA (retinoic acid receptor RXR-alpha), thereby inhibiting long-chain fatty acid β-oxidation by decreasing STAT3 (signal transducer and activator of transcription 3) expression and phosphorylation.ConclusionRisperidone increases adipocyte lipid accumulation by plausibly inhibiting long-chain fatty acid β-oxidation through targeting MAPK14 and MAPK8.</p

DataSheet1_Pathway Association Studies Reveal Gene Loci and Pathway Networks that Associated With Plasma Cystatin C Levels.docx

As a marker for glomerular filtration, plasma cystatin C level is used to evaluate kidney function. To decipher genetic factors that control the plasma cystatin C level, we performed genome-wide association and pathway association studies using United Kingdom Biobank data. One hundred fifteen loci yielded p values less than 1 × 10−100, three genes (clusters) showed the most significant associations, including the CST8-CST9 cluster on chromosome 20, the SH2B3-ATXN2 gene region on chromosome 12, and the SHROOM3-CCDC158 gene region on chromosome 4. In pathway association studies, forty significant pathways had FDR (false discovery rate) and or FWER (family-wise error rate) ≤ 0.001: spermatogenesis, leukocyte trans-endothelial migration, cell adhesion, glycoprotein, membrane lipid, steroid metabolic process, and insulin signaling pathways were among the most significant pathways that associated with the plasma cystatin C levels. We also performed Genome-wide association studies for eGFR, top associated genes were largely overlapped with those for cystatin C.</p

Induction of D-Lactate. Described the variation tendency of D-Lactate in two groups.

<p>Induction of D-Lactate. Described the variation tendency of D-Lactate in two groups.</p

Data of amylase, Cr (endogenous creatinine), TB (total bilirubin) and TNF-alpha for groups at each time point.

<p>In ANP and SO, ^a<i>P</i><0.05 vs 0 h;^b<i>P</i><0.05 vs 3 h; ^c<i>P</i><0.05 vs 6 h; ^d<i>P</i><0.05 vs 12 h.</p><p>There were significantly differences between ANP and SO at each time point.</p><p>Cr: endogenous creatinine, TB: total bilirubin.</p><p>ANP 3 h group n = 10, ANP 6 h group n = 8, ANP 12 h group n = 8, ANP 24 h group n = 6.</p><p>Data for each group at each time point (mean (SEM))</p

Induction of DAO. Described the variation tendency of DAO in two groups.

<p>Induction of DAO. Described the variation tendency of DAO in two groups.</p

Table_1_Deciphering Risperidone-Induced Lipogenesis by Network Pharmacology and Molecular Validation.docx

BackgroundRisperidone is an atypical antipsychotic that can cause substantial weight gain. The pharmacological targets and molecular mechanisms related to risperidone-induced lipogenesis (RIL) remain to be elucidated. Therefore, network pharmacology and further experimental validation were undertaken to explore the action mechanisms of RIL.MethodsRILs were systematically analyzed by integrating multiple databases through integrated network pharmacology, transcriptomics, molecular docking, and molecular experiment analysis. The potential signaling pathways for RIL were identified and experimentally validated using gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) analysis.ResultsRisperidone promotes adipocyte differentiation and lipid accumulation through Oil Red O staining and reverse transcription-polymerase chain reaction (RT-PCR). After network pharmacology and GO analysis, risperidone was found to influence cellular metabolism. In addition, risperidone influences adipocyte metabolism, differentiation, and lipid accumulation-related functions through transcriptome analysis. Intersecting analysis, molecular docking, and pathway validation analysis showed that risperidone influences the adipocytokine signaling pathway by targeting MAPK14 (mitogen-activated protein kinase 14), MAPK8 (mitogen-activated protein kinase 8), and RXRA (retinoic acid receptor RXR-alpha), thereby inhibiting long-chain fatty acid β-oxidation by decreasing STAT3 (signal transducer and activator of transcription 3) expression and phosphorylation.ConclusionRisperidone increases adipocyte lipid accumulation by plausibly inhibiting long-chain fatty acid β-oxidation through targeting MAPK14 and MAPK8.</p

Induction of IAP. Described the variation tendency of IAP in two groups.

<p>Induction of IAP. Described the variation tendency of IAP in two groups.</p

Data of weight of ascites, mortality, pathological scores of intestine and pancreas for groups at each time point.

<p>In ANP and SO, ^a<i>P</i><0.05 vs 0 h;^b<i>P</i><0.05 vs 3 h; ^c<i>P</i><0.05 vs 6 h; ^d<i>P</i><0.05 vs 12 h.</p><p>There were significantly differences between ANP and SO at each time point.</p><p>Data for each group at each time point (mean (SEM))</p

Construction of SDCBP silenced breast cancer cells.

<p>(<b>A</b>) Syndecan binding protein (SDCBP) and negative control (NC) short-hairpin RNA (shRNA) expression constructs were transiently transfected into human embryonic kidney 293T cells and the holoproteins in cell lysates were western-blotted for SDCBP shRNA selection. Digits 397, 523, 611 and 987 represent initiation site of four candidate target sequences selected in SDCBP mRNA (NM_001007067) and (*) represent the target sequence we selected as SDCBP shRNA. (<b>B</b>) MDA-MB-231 cells with SDCBP silenced were selected by Western Blot analysis. (*) represented the clone we selected as the MDA-MB-231-SDCBP shRNA cell. (<b>C</b>) BT-549 cells with SDCBP silenced were selected by Western Blot analysis. (*) represented the clone we selected as the BT-549-SDCBP shRNA cell.</p

Syndecan binding protein expression in normal breast and breast cancer tissue.

*<p>, the <i>P</i> value was calculated by Mann–Whitney <i>U</i>-test.</p