489f21e70f187d3017462a4d0b847e5f Lymph_Node_Molecular_Analysis.pdf 246d90ea862d778ec7d1af73ea9c8506a792ee8a Lymph_Node_Molecular_Analysis.pdf f5137a686953d04a7461d6ce23669a9a1d4978a64a9bc2d7abe81c9d183cfee9 Lymph_Node_Molecular_Analysis.pdf Title: Lymph Node Molecular Analysis with OSNA Enables the Identification of pT1 CRC Patients at Risk of Recurrence: A Multicentre Study Subject: Early-stage colorectal carcinoma (CRC)—pT1—is a therapeutic challenge and presents some histological features related to lymph node metastasis (LNM). A significant proportion of pT1 CRCs are treated surgically, resulting in a non-negligible surgical-associated mortality rate of 1.5–2%. Among these cases, approximately 6–16% exhibit LNM, but the impact on survival is unclear. Therefore, there is an unmet need to establish an objective and reliable lymph node (LN) staging method to optimise the therapeutic management of pT1 CRC patients and to avoid overtreating or undertreating them. In this multicentre study, 89 patients with pT1 CRC were included. All histological features associated with LNM were evaluated. LNs were assessed using two methods, One-Step Nucleic Acid Amplification (OSNA) and the conventional FFPE plus haematoxylin and eosin (H&E) staining. OSNA is an RT-PCR-based method for amplifying CK19 mRNA. Our aim was to assess the performance of OSNA and H&E in evaluating LNs to identify patients at risk of recurrence and to optimise their clinical management. We observed an 80.9% concordance in LN assessment using the two methods. In 9% of cases, LNs were found to be positive using H&E, and in 24.7% of cases, LNs were found to be positive using OSNA. The OSNA results are provided as the total tumour load (TTL), defined as the total tumour burden present in all the LNs of a surgical specimen. In CRC, a TTL 6000 CK19 m-RNA copies/L is associated with poor prognosis. Three patients had TTL > 6000 copies/L, which was associated with higher tumour budding. The discrepancies observed between the OSNA and H&E results were mostly attributed to tumour allocation bias. We concluded that LN assessment with OSNA enables the identification of pT1 CRC patients at some risk of recurrence and helps to optimise their clinical management. Keywords: pT1 colorectal cancer; lymph node; staging; diagnosis; OSNA Author: Karmele Saez de Gordoa, Maria Teresa Rodrigo-Calvo, Ivan Archilla, Sandra Lopez-Prades, Alba Diaz, Jordi Tarragona, Isidro Machado, Juan Ruiz Martín, Diana Zaffalon, Maria Daca-Alvarez, Maria Pellisé, Jordi Camps and Miriam Cuatrecasas Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Nov 21 08:44:43 2023 CET ModDate: Tue Nov 21 09:03:58 2023 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1913796 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- YSPQJI+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 DUFXDP+VnURWPalladioL Type 1 Custom yes yes no 15 0 FSIKDI+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 WBQRCQ+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 UQPKGM+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 47 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 69 0 OHXQCJ+CMSY10 Type 1 Builtin yes yes no 73 0 DECEIO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 93 0 DECEJA+PalatinoLinotype TrueType WinAnsi yes yes no 96 0 DECEKC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 99 0 FHBIAZ+EURB7 Type 1 Builtin yes yes no 106 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 110 0 DECEIO+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 121 0 DECEIP+PalatinoLinotype CID TrueType Identity-H yes yes yes 124 0 DECEJA+PalatinoLinotype TrueType WinAnsi yes yes no 130 0 DECEKC+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 133 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-03-21 02:36:21 CET RepresentationInformation: Lymph_Node_Molecular_Analysis.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-03-20 14:40:05 CET Size: 1913796 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 416 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 Samples Destination: subsection.2.1 Item: Title: Ethical Considerations Destination: subsection.2.2 Item: Title: Lymph Node Dissection and One-Step Nucleic Acid Amplification (OSNA) Destination: subsection.2.3 Item: Title: Lymph Node pN Staging and OSNA Results Destination: subsection.2.4 Item: Title: Histological Evaluation of pT1 CRC Destination: subsection.2.5 Item: Title: Statistical Analysis Destination: subsection.2.6 Item: Title: Results Destination: section.3 Children: Item: Title: Comparison of Lymph Node Status Assessed with H&E and OSNA Destination: subsection.3.1 Item: Title: Pathological Characteristics Related to Lymph Node Metastases Destination: subsection.3.2 Item: Title: Patient Follow-Up Destination: subsection.3.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.6 Info: Title: Lymph Node Molecular Analysis with OSNA Enables the Identification of pT1 CRC Patients at Risk of Recurrence: A Multicentre Study Author: Karmele Saez de Gordoa, Maria Teresa Rodrigo-Calvo, Ivan Archilla, Sandra Lopez-Prades, Alba Diaz, Jordi Tarragona, Isidro Machado, Juan Ruiz Martín, Diana Zaffalon, Maria Daca-Alvarez, Maria Pellisé, Jordi Camps and Miriam Cuatrecasas Subject: Early-stage colorectal carcinoma (CRC) pT1 is a therapeutic challenge and presents some histological features related to lymph node metastasis (LNM). A significant proportion of pT1 CRCs are treated surgically, resulting in a non-negligible surgical-associated mortality rate of 1.5 2%. Among these cases, approximately 6 16% exhibit LNM, but the impact on survival is unclear. Therefore, there is an unmet need to establish an objective and reliable lymph node (LN) staging method to optimise the therapeutic management of pT1 CRC patients and to avoid overtreating or undertreating them. In this multicentre study, 89 patients with pT1 CRC were included. All histological features associated with LNM were evaluated. LNs were assessed using two methods, One-Step Nucleic Acid Amplification (OSNA) and the conventional FFPE plus haematoxylin and eosin (H&E) staining. OSNA is an RT-PCR-based method for amplifying CK19 mRNA. Our aim was to assess the performance of OSNA and H&E in evaluating LNs to identify patients at risk of recurrence and to optimise their clinical management. We observed an 80.9% concordance in LN assessment using the two methods. In 9% of cases, LNs were found to be positive using H&E, and in 24.7% of cases, LNs were found to be positive using OSNA. The OSNA results are provided as the total tumour load (TTL), defined as the total tumour burden present in all the LNs of a surgical specimen. In CRC, a TTL 6000 CK19 m-RNA copies/L is associated with poor prognosis. Three patients had TTL > 6000 copies/L, which was associated with higher tumour budding. The discrepancies observed between the OSNA and H&E results were mostly attributed to tumour allocation bias. We concluded that LN assessment with OSNA enables the identification of pT1 CRC patients at some risk of recurrence and helps to optimise their clinical management. 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