fc35f4c37ddce68874ef299d6cf1cecb 299281.pdf d81a7e7d809d3ced83ba2d3dbef687c5d31d71ff 299281.pdf 324c19af88e2a03c064e227958faa709ecf7243b93badafadce9f119bf83e0bb 299281.pdf Title: Chromogenic LMO2 mRNA ISH Expression Correlates with LMO2 Protein and Gene Expression and Captures Their Survival Impact in Diffuse Large B-Cell Lymphoma, NOS Subject: Background: LMO2 is a relevant gene involved in B-cell ontogeny and a survival predictor of aggressive large B-cell lymphomas (aLBCL). Most studies assessing LMO2 mRNA expression have relied on microarray platforms or qRT-PCR methods, overlooking tissue morphology. In this study, we evaluate LMO2 RNA expression by chromogenic in situ hybridization (CISH) in normal tissue and in a series of 82 aLBCL. Methods: LMO2 CISH was performed in formalin-fixed paraffin-embedded tissues, scored by three different methods, and correlated with a transcriptome panel. Results: We obtained statistically significant results correlating the methods of evaluation with LMO2 protein expression and gene expression results. Normal tonsil tissue showed high levels of LMO2, particularly within the light zone of the germinal center. Conversely, in aLBCL, a notable reduction in LMO2 expression was noted, remarkably in cases carrying MYC rearrangements. Furthermore, significant results were obtained through overall survival and Cox regression survival analysis, incorporating International Prognostic Index data alongside LMO2 expression levels. Conclusions: We show a reliable method to identify LMO2 mRNA expression by CISH, effectively capturing many of the reported biologic features of LMO2. Keywords: LMO2; MYC; CISH; large B-cell lymphoma Author: Natalia Papaleo, Andrea Molina-Alvarez, Ricard Onieva, Diana Fuertes, Blanca Sanchez-Gonzalez, Xenia Riera, David Lopez-Segura, Carmen Lome-Maldonado, Xavier Ara-Mancebo, Jose Yelamos, Marta Salido, Ivonne Vazquez, Xavier Calvo and Luis Colomo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri Jun 28 12:19:39 2024 CEST ModDate: Fri Jun 28 12:22:37 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3934865 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- DNNUWA+VnURWPalladioL Type 1 Custom yes yes yes 10 0 ENFEZW+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 OMLVZU+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 JHTNQW+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 JFQMLV+URWPalladioL-BoldItal Type 1 Custom yes yes yes 32 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 49 0 DVCMRV+CMSY10 Type 1 Builtin yes yes yes 69 0 HLBFOA+DengXian CID TrueType Identity-H yes yes yes 80 0 HLBEIA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 86 0 HLBEIC+PalatinoLinotype TrueType WinAnsi yes yes no 89 0 HLBEIE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 92 0 HLBFHB+PalatinoLinotype TrueType MacRoman yes yes no 95 0 HLBEIA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 110 0 HLBEIB+PalatinoLinotype CID TrueType Identity-H yes yes yes 113 0 HLBEIC+PalatinoLinotype TrueType WinAnsi yes yes no 119 0 HLBEIE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 122 0 HLBFHB+PalatinoLinotype TrueType MacRoman yes yes no 125 0 HLBOEN+Arial,Bold CID TrueType Identity-H yes yes yes 136 0 HLBEIA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 142 0 HLBEIB+PalatinoLinotype CID TrueType Identity-H yes yes yes 145 0 HLBEIC+PalatinoLinotype TrueType WinAnsi yes yes no 151 0 HLBEIE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 154 0 HLBFFB+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 157 0 HLBFHB+PalatinoLinotype TrueType MacRoman yes yes no 160 0 HLBEIA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 172 0 HLBEIB+PalatinoLinotype CID TrueType Identity-H yes yes yes 175 0 HLBEIC+PalatinoLinotype TrueType WinAnsi yes yes no 181 0 HLBEIE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 184 0 HLBFHB+PalatinoLinotype TrueType MacRoman yes yes no 187 0 HLBEIA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 194 0 HLBEIB+PalatinoLinotype CID TrueType Identity-H yes yes yes 197 0 HLBEIC+PalatinoLinotype TrueType WinAnsi yes yes no 203 0 HLBEIE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 206 0 HLBFFB+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 209 0 HLBFHB+PalatinoLinotype TrueType MacRoman yes yes no 212 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-07-25 02:41:10 CEST RepresentationInformation: 299281.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-07-24 19:13:03 CEST Size: 3934865 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 365 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Case Selection Destination: subsection.2.1 Item: Title: Immunohistochemistry Destination: subsection.2.2 Item: Title: Fluorescence and Chromogenic In Situ Hybridization (FISH and CISH) Destination: subsection.2.3 Item: Title: Gene Expression Analysis Destination: subsection.2.4 Item: Title: Statistical Analysis Destination: subsection.2.5 Item: Title: Results Destination: section.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.6 Info: Title: Chromogenic LMO2 mRNA ISH Expression Correlates with LMO2 Protein and Gene Expression and Captures Their Survival Impact in Diffuse Large B-Cell Lymphoma, NOS Author: Natalia Papaleo, Andrea Molina-Alvarez, Ricard Onieva, Diana Fuertes, Blanca Sanchez-Gonzalez, Xenia Riera, David Lopez-Segura, Carmen Lome-Maldonado, Xavier Ara-Mancebo, Jose Yelamos, Marta Salido, Ivonne Vazquez, Xavier Calvo and Luis Colomo Subject: Background: LMO2 is a relevant gene involved in B-cell ontogeny and a survival predictor of aggressive large B-cell lymphomas (aLBCL). Most studies assessing LMO2 mRNA expression have relied on microarray platforms or qRT-PCR methods, overlooking tissue morphology. In this study, we evaluate LMO2 RNA expression by chromogenic in situ hybridization (CISH) in normal tissue and in a series of 82 aLBCL. Methods: LMO2 CISH was performed in formalin-fixed paraffin-embedded tissues, scored by three different methods, and correlated with a transcriptome panel. Results: We obtained statistically significant results correlating the methods of evaluation with LMO2 protein expression and gene expression results. Normal tonsil tissue showed high levels of LMO2, particularly within the light zone of the germinal center. Conversely, in aLBCL, a notable reduction in LMO2 expression was noted, remarkably in cases carrying MYC rearrangements. Furthermore, significant results were obtained through overall survival and Cox regression survival analysis, incorporating International Prognostic Index data alongside LMO2 expression levels. Conclusions: We show a reliable method to identify LMO2 mRNA expression by CISH, effectively capturing many of the reported biologic features of LMO2. 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