f38be360510696127c43546a207072fd pmc_9913321.pdf e8f8fb552c996cebd8d93122044a6cfb5855f96d pmc_9913321.pdf ae2c922528f836c46e4b78a72d3c465c1f124b5a7ed0ae360fea398228499e91 pmc_9913321.pdf Title: Tumour Cell Seeding to Lymph Nodes from In Situ Colorectal Cancer Subject: Lymph node (LN) metastasis is an important prognostic factor in colorectal cancer (CRC). We aimed to demonstrate the presence of lymphatic vessels (LV) in the mucosa of in-situ (pTis) CRC, and of detectable tumour burden in regional LNs. This is an observational retrospective study of 39 surgically resected in situ CRCs. The number of LVs was evaluated in both pTis and normal mucosa using D2-40 immunostains. All LNs were assessed with both H&E and the One Step Nucleic Acid Amplification (OSNA) assay, and the results were correlated with clinicopathological features. D2-40 immunohistochemisty revealed LVs in the lamina propria of all pTis CRC (100%), being absent in normal mucosa. A median of 16 LNs were freshly dissected per patient, and all cases were pN0 with H&E. Molecular LN analysis with OSNA revealed the presence of low amounts of tumour burden in 11/39 (28%) cases (range 400 to 4270 CK19 mRNA copies/L), which had no clinical consequences. This study demonstrates the presence of LVs in the lamina propria in 100% of pTis CRC, as well as the presence of low amounts of tumour burden in regional LNs, only detected by molecular methods. Given the prognostic value of LN tumour burden, its molecular quantification may help a patient’s clinical management. Keywords: in situ colorectal cancer; lymph node; staging; diagnosis; OSNA Author: Maria Teresa Rodrigo-Calvo, Karmele Saez de Gordoa, Sandra Lopez-Prades, Ivan Archilla, Alba Diaz, Mario Berrios, Jordi Camps, Eva Musulen and Miriam Cuatrecasas Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Jan 30 04:55:51 2023 CET ModDate: Mon Jan 30 05:00:47 2023 CET 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: 1709321 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- YFNERV+VnURWPalladioL Type 1 Custom yes yes no 10 0 RFECCF+EURM10 Type 1 Builtin yes yes no 15 0 GIFMJF+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 FETICY+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 AGQWCB+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 47 0 VRITEV+CMSY10 Type 1 Builtin yes yes no 72 0 CNDCIH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 89 0 CNDCII+PalatinoLinotype CID TrueType Identity-H yes yes yes 92 0 CNDCIJ+PalatinoLinotype TrueType WinAnsi yes yes no 98 0 CNDCIL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 101 0 CNDEHC+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 104 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 111 0 CNDCIH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 122 0 CNDCII+PalatinoLinotype CID TrueType Identity-H yes yes yes 125 0 CNDCIJ+PalatinoLinotype TrueType WinAnsi yes yes no 131 0 CNDCIL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 134 0 CNDEHC+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 137 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-01-19 04:07:00 CET RepresentationInformation: pmc_9913321.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-30 02:16:06 CEST Size: 1709321 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 292 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: Patients and Tumours Destination: subsection.2.1 Item: Title: Ethical Considerations Destination: subsection.2.2 Item: Title: Immunohistochemistry Staining Destination: subsection.2.3 Item: Title: Lymphatic Vessel Assessment Destination: subsection.2.4 Item: Title: Intratumour Budding Evaluation Destination: subsection.2.5 Item: Title: Lymph Node Dissection and Analysis Destination: subsection.2.6 Item: Title: One Step Nucleic Acid Amplification (OSNA) Assay Destination: subsection.2.7 Item: Title: Lymph Node Staging Destination: subsection.2.8 Item: Title: Statistical Analysis Destination: subsection.2.9 Item: Title: Results Destination: section.3 Children: Item: Title: Study Sample Destination: subsection.3.1 Item: Title: The Normal Colonic Mucosa Is Devoid of Lymphatic Vessels Destination: subsection.3.2 Item: Title: Presence of Lymphatic Vessels in the Mucosa of In Situ CRC Destination: subsection.3.3 Item: Title: Intratumoural Tumour Budding in pTis Destination: subsection.3.4 Item: Title: Detection of Tumour Burden in Regional LNs with the OSNA Assay Destination: subsection.3.5 Item: Title: Patient’s Follow-Up Destination: subsection.3.6 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: Tumour Cell Seeding to Lymph Nodes from In Situ Colorectal Cancer Author: Maria Teresa Rodrigo-Calvo, Karmele Saez de Gordoa, Sandra Lopez-Prades, Ivan Archilla, Alba Diaz, Mario Berrios, Jordi Camps, Eva Musulen and Miriam Cuatrecasas Subject: Lymph node (LN) metastasis is an important prognostic factor in colorectal cancer (CRC). We aimed to demonstrate the presence of lymphatic vessels (LV) in the mucosa of in-situ (pTis) CRC, and of detectable tumour burden in regional LNs. This is an observational retrospective study of 39 surgically resected in situ CRCs. The number of LVs was evaluated in both pTis and normal mucosa using D2-40 immunostains. All LNs were assessed with both H&E and the One Step Nucleic Acid Amplification (OSNA) assay, and the results were correlated with clinicopathological features. D2-40 immunohistochemisty revealed LVs in the lamina propria of all pTis CRC (100%), being absent in normal mucosa. A median of 16 LNs were freshly dissected per patient, and all cases were pN0 with H&E. Molecular LN analysis with OSNA revealed the presence of low amounts of tumour burden in 11/39 (28%) cases (range 400 to 4270 CK19 mRNA copies/L), which had no clinical consequences. This study demonstrates the presence of LVs in the lamina propria in 100% of pTis CRC, as well as the presence of low amounts of tumour burden in regional LNs, only detected by molecular methods. Given the prognostic value of LN tumour burden, its molecular quantification may help a patient’s clinical management. 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