879b9b5f81e4bc2d6caf5667d365a536 ijms-25-06437.pdf fb6488bfa53a993c5b8f83b3c8d67538aec85cec ijms-25-06437.pdf 1634600f958c514346aa0360f950a81d7e1ad0306bf709296fe07cb753d8fa12 ijms-25-06437.pdf Title: Integrated miRNA–mRNA Analysis Reveals Critical miRNAs and Targets in Diet-Induced Obesity-Related Glomerulopathy Subject: This study aimed to investigate obesity-related glomerulopathy (ORG) at cellular, structural, and transcriptomic levels. Thirty Wistar rats were randomized into two groups: 15 rats were fed with a standard diet (SD-rats), and 15 rats were fed with a high-fat diet (HFD-rats). After 10 weeks, the weight, kidney function, histological features, and transcriptomic changes were assessed. HFD-rats gained significantly more weight (55.8% vs. 29.2%; p < 0.001) and albuminuria (10,384.04 ng/mL vs. 5845.45 ng/mL; p < 0.001) compared to SD-rats. HFD-rats exhibited early stages of ORG, with predominant mesangial matrix increase and podocyte hypertrophy (PH). These lesions correlated with differentially expressed (DE) genes and miRNAs. Functional analysis showed that miR-205, which was DE in both the kidneys and urine of HFD-rats, negatively regulated the PTEN gene, promoting lipid endocytosis in podocytes. The downregulation of PTEN was proved through a higher PTEN/nephrin ratio in the SD-rats and the presence of lipid vacuoles in HFD-podocytes. This study has found a specific targetome of miRNAs and gene expression in early stages of ORG. Also, it emphasizes the potential value of miR-205 as a urinary biomarker for detecting podocyte injury in ORG, offering a tool for early diagnosis, and opening new avenues for future therapeutic research of obesity-related glomerulopathy. Keywords: microRNA; biomarker; targetome; obesity-related glomerulopathy; mesangial matrix increase; podocyte hypertrophy; Wistar rats Author: Marina López-Martínez, Maria Pilar Armengol, Irina Pey, Xavier Farré, Paula Rodríguez-Martínez, Mireia Ferrer, Esteban Porrini, Sergio Luis-Lima, Laura Díaz-Martín, Ana Elena Rodríguez-Rodríguez, Coriolano Cruz-Perera, Marta Alcalde and Maruja Navarro-Díaz Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Tue Jun 11 15:19:50 2024 CEST ModDate: Tue Jun 11 15:24:42 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 21 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 7434408 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 OTYBJA+URWPalladioL-Bold Type 1 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Destination: subsection.2.1 Item: Title: Renal Function Destination: subsection.2.2 Item: Title: Study of Renal Morphological Changes Destination: subsection.2.3 Item: Title: Diet-Induced Transcriptomic Changes and Validation of DE RNAs in Rat Kidneys Destination: subsection.2.4 Item: Title: Lesion-Related Transcriptomic Changes and DE RNAs in Rat Kidneys Destination: subsection.2.5 Item: Title: Small RNA Profile Changes According to Diet and Weight Destination: subsection.2.6 Item: Title: Functional Analysis of Pathways and Leading-Edge Genes Targeted by DE miRNAs Destination: subsection.2.7 Item: Title: Lesion-Related Transcriptomic Changes in Rat Kidneys (small RNA) Destination: subsection.2.8 Item: Title: Diet-Dependent Expressions of Identified Small RNAs in Rat Urine Samples Destination: subsection.2.9 Item: Title: Lesion-Related Transcriptomic Changes in Urine Destination: subsection.2.10 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Establishment of the Obesity-Induced Animal Model Destination: subsection.4.1 Item: Title: Organ and Urine Harvesting Destination: subsection.4.2 Item: Title: Measurement of Renal Function: Albuminuria and Iohexol Clearance Destination: subsection.4.3 Item: Title: Morphometry, Histology, Immunohistochemistry, Immunofluorescence, and Transmission Electron Microscopy (TEM) Destination: subsection.4.4 Item: Title: Total RNA Isolation and miRNA Enrichment Destination: subsection.4.5 Item: Title: Library Preparation, RNA Sequencing, and Data Analysis Destination: subsection.4.6 Item: Title: Library Preparation, smallRNA Sequencing, and Data Analysis Destination: subsection.4.7 Item: Title: Predicted and Validated Targetome for miRNAs Destination: subsection.4.8 Item: Title: Gene ontology and Pathway Analyses Destination: subsection.4.9 Item: Title: Reverse Transcription and Quantitative Reverse Transcription Polymerase Chain Reaction Validation Destination: subsection.4.10 Item: Title: Statistical Analysis Destination: subsection.4.11 Item: Title: Study Approval Destination: subsection.4.12 Item: Title: References Destination: appendix.A. Info: Title: Integrated miRNA mRNA Analysis Reveals Critical miRNAs and Targets in Diet-Induced Obesity-Related Glomerulopathy Author: Marina López-Martínez, Maria Pilar Armengol, Irina Pey, Xavier Farré, Paula Rodríguez-Martínez, Mireia Ferrer, Esteban Porrini, Sergio Luis-Lima, Laura Díaz-Martín, Ana Elena Rodríguez-Rodríguez, Coriolano Cruz-Perera, Marta Alcalde and Maruja Navarro-Díaz Subject: This study aimed to investigate obesity-related glomerulopathy (ORG) at cellular, structural, and transcriptomic levels. Thirty Wistar rats were randomized into two groups: 15 rats were fed with a standard diet (SD-rats), and 15 rats were fed with a high-fat diet (HFD-rats). After 10 weeks, the weight, kidney function, histological features, and transcriptomic changes were assessed. HFD-rats gained significantly more weight (55.8% vs. 29.2%; p < 0.001) and albuminuria (10,384.04 ng/mL vs. 5845.45 ng/mL; p < 0.001) compared to SD-rats. HFD-rats exhibited early stages of ORG, with predominant mesangial matrix increase and podocyte hypertrophy (PH). These lesions correlated with differentially expressed (DE) genes and miRNAs. Functional analysis showed that miR-205, which was DE in both the kidneys and urine of HFD-rats, negatively regulated the PTEN gene, promoting lipid endocytosis in podocytes. The downregulation of PTEN was proved through a higher PTEN/nephrin ratio in the SD-rats and the presence of lipid vacuoles in HFD-podocytes. This study has found a specific targetome of miRNAs and gene expression in early stages of ORG. Also, it emphasizes the potential value of miR-205 as a urinary biomarker for detecting podocyte injury in ORG, offering a tool for early diagnosis, and opening new avenues for future therapeutic research of obesity-related glomerulopathy. Keywords: microRNA; biomarker; targetome; obesity-related glomerulopathy; mesangial matrix increase; podocyte hypertrophy; Wistar rats Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Tue Jun 11 15:19:50 CEST 2024 ModDate: Tue Jun 11 15:24:42 CEST 2024 ID: 0xe07d4ebca23cdaf05acc0bf1b7849af4, 0xe07d4ebca23cdaf05acc0bf1b7849af4 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: XUDAJP+VnURWPalladioL FontSubset: true FirstChar: 128 LastChar: 243 FontDescriptor: FontName: XUDAJP+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: OTYBJA+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 150 FontDescriptor: FontName: OTYBJA+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: BBSSAI+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 241 FontDescriptor: 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