e2accf28a59bc92c1311fc96412513a9 can_a2021m6v13n11p2663.pdf 04de5223c593fb875306ab02ca0d58fc642318dc can_a2021m6v13n11p2663.pdf 747084f0cc01103093edcf8b586cc700de57114d6c08b3049d6401fff7608a8f can_a2021m6v13n11p2663.pdf Title: Immune System-Related Changes in Preclinical GL261 Glioblastoma under TMZ Treatment: Explaining MRSI-Based Nosological Imaging Findings with RT-PCR Analyses Subject: Glioblastomas (GB) are brain tumours with poor prognosis even after aggressive therapy. Previous work suggests that magnetic resonance spectroscopic imaging (MRSI) could act as a biomarker of efficient immune system attack onto GB, presenting oscillatory changes. Glioma-associated microglia/macrophages (GAMs) constitute the most abundant non-tumour cell type within the GB and can be polarised into anti-tumour (M1) or pro-tumour (M2) phenotypes. One of the mechanisms to mediate immunosuppression in brain tumours is the interaction between programmed cell death-1 ligand 1 (PD-L1) and programmed cell death-1 receptor (PD-1). We evaluated the subpopulations of GAMs in responding and control GB tumours to correlate PD-L1 expression to GAM polarisation in order to explain/validate MRSI-detected findings. Mice were evaluated by MRI/MRSI to assess the extent of response to treatment and with qPCR for GAMs M1 and M2 polarisation analyses. M1/M2 ratios and PD-L1 expression were higher in treated compared to control tumours. Furthermore, PD-L1 expression was positively correlated with the M1/M2 ratio. The oscillatory change in the GAMs prevailing population could be one of the key causes for the differential MRSI-detected pattern, allowing this to act as immune system activity biomarker in future work. Keywords: glioblastoma; orthotopic immunocompetent tumours; immune-enhancing metronomic schedule; TMZ; magnetic resonance spectroscopic imaging; immune system activity imaging biomarker; cancer immune cycle; glioma-associated microglia/macrophages; PD-L1 Author: Pilar Calero-Pérez, Shuang Wu, Carles Arús and Ana Paula Candiota Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Jun 7 02:47:18 2021 CEST ModDate: Tue Jul 13 17:56:42 2021 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1408206 bytes Optimized: yes PDF version: 1.6 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- MQFYIK+VnURWPalladioL-Bold Type 1 Custom yes yes no 482 0 NQWDWO+VnURWPalladioL Type 1 Custom yes yes no 483 0 CDXJMD+URWPalladioL-Ital Type 1 Custom yes yes no 484 0 QIHDIK+URWPalladioL-Bold Type 1 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yes yes no 118 0 DMBMOH+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 119 0 DMBMOD+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 134 0 DMBMOF+PalatinoLinotype TrueType WinAnsi yes yes no 135 0 DMBMOH+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 136 0 DMBMOD+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 163 0 DMBMOF+PalatinoLinotype TrueType WinAnsi yes yes no 164 0 DMBMOH+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 165 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2021-07-14 02:29:31 CEST RepresentationInformation: can_a2021m6v13n11p2663.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2021-07-13 18:13:44 CEST Size: 1408206 Format: PDF Version: 1.6 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf Profile: Linearized PDF PDFMetadata: Objects: 545 FreeObjects: 1 IncrementalUpdates: 1 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Children: Item: Title: GL261 GB Preclinical Model Generation and Treatment Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: In Vivo MRI and MRSI Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Children: Item: Title: MRI Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: MRSI Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: MRI and MRSI Processing and Post-Processing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Tumour Responding index (TRI) Calculations Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Animal Euthanasia Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: RNA Isolation, cDNA Synthesis and qPCR Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Children: Item: Title: Follow up of GL261 Tumour-Bearing Mice and Endpoint Criteria Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Item: Title: Microglia/Macrophage Global Population, As Well As M1 and M2 Subtypes, Are Increased in IMS-TMZ-Treated Tumours Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2 Item: Title: Assessing Different Macrophage Population Subtypes Regarding Global GAMs Values Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@491cc5c9 Item: Title: PD-L1 Gene Is Highly Expressed in IMS-TMZ-Treated Tumours, and These Increases May Be Correlated with the Polarisation State of Microglia/Macrophage Population Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@74ad1f1f Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6a1aab78 Children: Item: Title: Immune System Populations Change during Response to Therapy Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@462d5aee Item: Title: Not All GAMs Are Equal (I): Polarisation of Microglia/Macrophages and Its Role in Therapy Response in GB Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@69b0fd6f Item: Title: Not All GAMs Are Equal (II): The Polarisation Status of Microglia/Macrophages Affects Their Metabolomic Pattern Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@757942a1 Item: Title: PD-L1 Gene Expression in GL261 GB Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4a87761d Item: Title: Wrap-up: Incoporating the Measured Gene Expression Results into the Explanation of the Oscillatory Behaviour of the MRSI-Based Biomarker of Response Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@66d1af89 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@8646db9 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@37374a5e Info: Title: Immune System-Related Changes in Preclinical GL261 Glioblastoma under TMZ Treatment: Explaining MRSI-Based Nosological Imaging Findings with RT-PCR Analyses Author: Pilar Calero-Pérez, Shuang Wu, Carles Arús and Ana Paula Candiota Subject: Glioblastomas (GB) are brain tumours with poor prognosis even after aggressive therapy. Previous work suggests that magnetic resonance spectroscopic imaging (MRSI) could act as a biomarker of efficient immune system attack onto GB, presenting oscillatory changes. Glioma-associated microglia/macrophages (GAMs) constitute the most abundant non-tumour cell type within the GB and can be polarised into anti-tumour (M1) or pro-tumour (M2) phenotypes. One of the mechanisms to mediate immunosuppression in brain tumours is the interaction between programmed cell death-1 ligand 1 (PD-L1) and programmed cell death-1 receptor (PD-1). We evaluated the subpopulations of GAMs in responding and control GB tumours to correlate PD-L1 expression to GAM polarisation in order to explain/validate MRSI-detected findings. Mice were evaluated by MRI/MRSI to assess the extent of response to treatment and with qPCR for GAMs M1 and M2 polarisation analyses. M1/M2 ratios and PD-L1 expression were higher in treated compared to control tumours. Furthermore, PD-L1 expression was positively correlated with the M1/M2 ratio. The oscillatory change in the GAMs prevailing population could be one of the key causes for the differential MRSI-detected pattern, allowing this to act as immune system activity biomarker in future work. Keywords: glioblastoma; orthotopic immunocompetent tumours; immune-enhancing metronomic schedule; TMZ; magnetic resonance spectroscopic imaging; immune system activity imaging biomarker; cancer immune cycle; glioma-associated microglia/macrophages; PD-L1 Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Jun 07 02:47:18 CEST 2021 ModDate: Tue Jul 13 17:56:42 CEST 2021 ID: 0x916f5d754454df66d646e0c64ab1f5b1, 0x0ae3c63f95f1f44394bb50b941bd3c36 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: MQFYIK+VnURWPalladioL-Bold FontSubset: true FirstChar: 178 LastChar: 243 FontDescriptor: FontName: MQFYIK+VnURWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: NQWDWO+VnURWPalladioL FontSubset: true FirstChar: 160 LastChar: 243 FontDescriptor: FontName: NQWDWO+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: PQVFRT+CMSY10 FontSubset: true FirstChar: 0 LastChar: 20 FontDescriptor: FontName: PQVFRT+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true Font: BaseFont: CDXJMD+URWPalladioL-Ital FontSubset: true FirstChar: 39 LastChar: 151 FontDescriptor: FontName: CDXJMD+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: IDSVYB+VnURWPalladioL-Italic FontSubset: true FirstChar: 145 LastChar: 226 FontDescriptor: FontName: IDSVYB+VnURWPalladioL-Italic Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: QIHDIK+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 122 FontDescriptor: FontName: QIHDIK+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: FTPKOQ+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: FTPKOQ+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: OVNEXW+EURM10 FontSubset: true FirstChar: 22 LastChar: 22 FontDescriptor: FontName: OVNEXW+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: JHYTSG+CMR10 FontSubset: true FirstChar: 61 LastChar: 61 FontDescriptor: FontName: JHYTSG+CMR10 Flags: Symbolic FontBBox: -40, -250, 1009, 750 FontFile: true Font: BaseFont: MPUDFL+PazoMath FontSubset: true FirstChar: 68 LastChar: 68 FontDescriptor: FontName: MPUDFL+PazoMath Flags: Symbolic FontBBox: -40, -283, 878, 946 FontFile: true XMP: 2021-06-07T08:47:18+08:00 LaTeX with hyperref 2021-07-13T17:56:42+02:00 2021-07-13T17:56:42+02:00 application/pdf Glioblastomas (GB) are brain tumours with poor prognosis even after aggressive therapy. Previous work suggests that magnetic resonance spectroscopic imaging (MRSI) could act as a biomarker of efficient immune system attack onto GB, presenting oscillatory changes. Glioma-associated microglia/macrophages (GAMs) constitute the most abundant non-tumour cell type within the GB and can be polarised into anti-tumour (M1) or pro-tumour (M2) phenotypes. One of the mechanisms to mediate immunosuppression in brain tumours is the interaction between programmed cell death-1 ligand 1 (PD-L1) and programmed cell death-1 receptor (PD-1). We evaluated the subpopulations of GAMs in responding and control GB tumours to correlate PD-L1 expression to GAM polarisation in order to explain/validate MRSI-detected findings. Mice were evaluated by MRI/MRSI to assess the extent of response to treatment and with qPCR for GAMs M1 and M2 polarisation analyses. M1/M2 ratios and PD-L1 expression were higher in treated compared to control tumours. Furthermore, PD-L1 expression was positively correlated with the M1/M2 ratio. The oscillatory change in the GAMs prevailing population could be one of the key causes for the differential MRSI-detected pattern, allowing this to act as immune system activity biomarker in future work. Pilar Calero-Pérez, Shuang Wu, Carles Arús and Ana Paula Candiota Immune System-Related Changes in Preclinical GL261 Glioblastoma under TMZ Treatment: Explaining MRSI-Based Nosological Imaging Findings with RT-PCR Analyses glioblastoma; orthotopic immunocompetent tumours; immune-enhancing metronomic schedule; TMZ; magnetic resonance spectroscopic imaging; immune system activity imaging biomarker; cancer immune cycle; glioma-associated microglia/macrophages; PD-L1 pdfTeX-1.40.21 uuid:92d57cc3-6592-4a5b-86ef-c42d28578097 uuid:86051a5e-1d74-4f1c-8a39-1d390cb21000 Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 35, 759, 153, 795 Annotation: Subtype: Link Rect: 515, 759, 560, 795 Annotation: Subtype: Link Rect: 273, 641, 284, 654 Annotation: Subtype: Link Rect: 422, 641, 433, 654 Annotation: Subtype: Link Rect: 107, 346, 154, 355 Annotation: Subtype: Link Rect: 35, 334, 140, 343 Annotation: Subtype: Link Rect: 35, 162, 97, 184 Annotation: Subtype: Link Rect: 153, 89, 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