46ca7961b2e4e999b3a9a999aa7e0fb8 pmc_10037622.pdf bf2de2fcf19448231184da8803e6c3510d31617c pmc_10037622.pdf df97d1c59b61a27971f5e5c8aef1ec90d062d193e7fd406927b1146dbbc6839e pmc_10037622.pdf Title: Immunoregulatory Biomarkers of the Remission Phase in Type 1 Diabetes: miR-30d-5p Modulates PD-1 Expression and Regulatory T Cell Expansion Subject: The partial remission (PR) phase of type 1 diabetes (T1D) is an underexplored period characterized by endogenous insulin production and downmodulated autoimmunity. To comprehend the mechanisms behind this transitory phase and develop precision medicine strategies, biomarker discovery and patient stratification are unmet needs. MicroRNAs (miRNAs) are small RNA molecules that negatively regulate gene expression and modulate several biological processes, functioning as biomarkers for many diseases. Here, we identify and validate a unique miRNA signature during PR in pediatric patients with T1D by employing small RNA sequencing and RT-qPCR. These miRNAs were mainly related to the immune system, metabolism, stress, and apoptosis pathways. The implication in autoimmunity of the most dysregulated miRNA, miR-30d-5p, was evaluated in vivo in the non-obese diabetic mouse. MiR-30d-5p inhibition resulted in increased regulatory T cell percentages in the pancreatic lymph nodes together with a higher expression of CD200. In the spleen, a decrease in PD-1+ T lymphocytes and reduced PDCD1 expression were observed. Moreover, miR-30d-5p inhibition led to an increased islet leukocytic infiltrate and changes in both effector and memory T lymphocytes. In conclusion, the miRNA signature found during PR shows new putative biomarkers and highlights the immunomodulatory role of miR-30d-5p, elucidating the processes driving this phase. Keywords: microRNA; type 1 diabetes; biomarker; honeymoon; partial remission; childhood; immune regulation; epigenetics; islet autoimmunity; regulatory T cell Author: Laia Gomez-Muñoz, David Perna-Barrull, Marta Murillo, Maria Pilar Armengol, Marta Alcalde, Marti Catala, Silvia Rodriguez-Fernandez, Sergi Sunye, Aina Valls, Jacobo Perez, Raquel Corripio and Marta Vives-Pi Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Feb 28 09:46:16 2023 CET ModDate: Tue Feb 28 10:03:52 2023 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 27 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 6483195 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- DUFXDP+VnURWPalladioL Type 1 Custom yes yes no 10 0 SQGFTX+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 EEYOKW+URWPalladioL-Roma Type 1 Custom yes yes no 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WinAnsi yes yes no 283 0 IMCNAB+PalatinoLinotype CID TrueType Identity-H yes yes yes 286 0 IMCNAC+PalatinoLinotype TrueType WinAnsi yes yes no 292 0 IMCNAE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 295 0 IMCMPA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 310 0 IMCNAB+PalatinoLinotype CID TrueType Identity-H yes yes yes 313 0 IMCNAC+PalatinoLinotype TrueType WinAnsi yes yes no 319 0 IMCNAE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 322 0 XIRVNA+EURB10 Type 1 Builtin yes yes no 351 0 CRUZTT+VnURWPalladioL-Italic Type 1 Custom yes yes no 370 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-08-18 02:29:08 CEST RepresentationInformation: pmc_10037622.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-07-26 02:15:09 CEST Size: 6483195 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 662 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Distinct miRNA Signature during the Partial Remission Phase of Pediatric Patients with Type 1 Diabetes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Validation of Differentially Expressed miRNAs during the Partial Remission Phase Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Potential Enriched Biological Functions and Pathways under the Regulation of Differentially Expressed miRNAs during the Partial Remission Phase Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: In Vivo Inhibition of miR-30d-5p Potentiates Treg Expansion by Increasing CD200 Levels Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: In Vivo Inhibition of miR-30d-5p Reduces PD-1 Expression on Splenocytes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: In Vivo Inhibition of miR-30d-5p Displays Changes in Additional T Cell Differentiation Subsets Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: In Vivo Inhibition of miR-30d-5p Tends to Increase Leukocyte Islet Infiltration Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Children: Item: Title: Human Sample Collection and T1D Remission Follow-Up Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Clinical and Laboratory Testing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Total RNA Isolation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: RNA Library Preparation, Sequencing, and Data Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Gene Targets for miRNAs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Gene Ontology and Pathway Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Quantitative RT-qPCR Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: In Vivo miR-30d-5p Inhibitor Administration Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Insulitis Score Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Flow Cytometry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Children: Item: Title: Leukocyte Isolation from Spleen and PLN Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Viability and Cell Counting Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Immunophenotype Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Ethics Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Children: Item: Title: Human Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Item: Title: Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@491cc5c9 Info: Title: Immunoregulatory Biomarkers of the Remission Phase in Type 1 Diabetes: miR-30d-5p Modulates PD-1 Expression and Regulatory T Cell Expansion Author: Laia Gomez-Muñoz, David Perna-Barrull, Marta Murillo, Maria Pilar Armengol, Marta Alcalde, Marti Catala, Silvia Rodriguez-Fernandez, Sergi Sunye, Aina Valls, Jacobo Perez, Raquel Corripio and Marta Vives-Pi Subject: The partial remission (PR) phase of type 1 diabetes (T1D) is an underexplored period characterized by endogenous insulin production and downmodulated autoimmunity. To comprehend the mechanisms behind this transitory phase and develop precision medicine strategies, biomarker discovery and patient stratification are unmet needs. MicroRNAs (miRNAs) are small RNA molecules that negatively regulate gene expression and modulate several biological processes, functioning as biomarkers for many diseases. Here, we identify and validate a unique miRNA signature during PR in pediatric patients with T1D by employing small RNA sequencing and RT-qPCR. These miRNAs were mainly related to the immune system, metabolism, stress, and apoptosis pathways. The implication in autoimmunity of the most dysregulated miRNA, miR-30d-5p, was evaluated in vivo in the non-obese diabetic mouse. MiR-30d-5p inhibition resulted in increased regulatory T cell percentages in the pancreatic lymph nodes together with a higher expression of CD200. In the spleen, a decrease in PD-1+ T lymphocytes and reduced PDCD1 expression were observed. Moreover, miR-30d-5p inhibition led to an increased islet leukocytic infiltrate and changes in both effector and memory T lymphocytes. In conclusion, the miRNA signature found during PR shows new putative biomarkers and highlights the immunomodulatory role of miR-30d-5p, elucidating the processes driving this phase. Keywords: microRNA; type 1 diabetes; biomarker; honeymoon; partial remission; childhood; immune regulation; epigenetics; islet autoimmunity; regulatory T cell Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Feb 28 09:46:16 CET 2023 ModDate: Tue Feb 28 10:03:52 CET 2023 ID: 0x0fda363c2236020c19f227d61aff3daf, 0x0fda363c2236020c19f227d61aff3daf Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: MPUDFL+PazoMath FontSubset: true FirstChar: 68 LastChar: 68 FontDescriptor: FontName: MPUDFL+PazoMath Flags: Symbolic FontBBox: -40, -283, 878, 946 FontFile: true Font: BaseFont: CRUZTT+VnURWPalladioL-Italic FontSubset: true FirstChar: 189 LastChar: 189 FontDescriptor: FontName: CRUZTT+VnURWPalladioL-Italic Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: EEYOKW+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: EEYOKW+URWPalladioL-Roma Flags: Symbolic 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Type: CRC32 Checksum: 46ca7961b2e4e999b3a9a999aa7e0fb8 Type: MD5 Checksum: bf2de2fcf19448231184da8803e6c3510d31617c Type: SHA-1