Serial H&E (A,C) and BAP1 IHC (B, D) stained sections of pancreatic ductal adenocarcinomas which demonstrate positive immunohistochemical staining for BAP1.

<p>All the neoplastic and non-neoplastic cell demonstrate diffuse strong nuclear staining (Original magnification A,B 100x, C,D 400x).</p

Clinical and pathological characteristics of 306 pancreatic adenocarcinoma patients.

<p>Clinical and pathological characteristics of 306 pancreatic adenocarcinoma patients.</p

Haematoxylin & eosin (A, C) and BAP1 immunohistochemistry (B, D) stained sections from the sole pancreatic ductal adenocarcinoma patient who expressed loss of nuclear BAP1.

<p>In this case, the exocrine cells (solid arrows) clearly lacked the brown BAP1 staining, and the non-neoplastic endothelial cells serve as the internal positive controls (hollow arrows). Magnifications: A, B 100x; C, D 400x.</p

Kaplan-Meier survival curve for 306 pancreatic ductal adenocarcinoma patients.

<p>Kaplan-Meier survival curve for 306 pancreatic ductal adenocarcinoma patients.</p

Patient demographics.

BackgroundNext generation sequencing (NGS) is increasingly used in standard clinical practice to identify patients with potentially actionable mutations. Stratification of NGS mutation tiers is currently based on the European Society of Medical Oncology (ESMO) Scale for Clinical Actionability of Molecular Targets (ESCAT[E]) Tier I–V & X. Allele frequency is also increasingly recognised as an important prognostic tool in advanced cancer. The aim of this study was to determine the genomic mutations in metastatic colorectal cancer (CRC) in an Australian multicultural population and their influence on survival outcomes.MethodsNext generation sequencing with the 50-gene panel Oncomine Precision Assay™ was used on 180 CRC tissue samples obtained across six Sydney hospitals between June 2021 and March 2022.ResultsFrom 180 samples, 147 (82%) had at least one gene mutation identified with 68 (38%) having two or more concurrent mutations. Tier I variants included RAS wild-type [EI] in 73 (41%) and BRAF V600E [EIA] in 27 (15%). Non-tier I variants include 2 (1%) ERBB2 amplification [EIIB], 26 (15%) PIK3CA hotspot mutations [EIIIA] and 9 (5%) MET focal amplifications [EIIIA]. NGS testing revealed an additional 22% of cases with Tier II & III mutations. 43% of patients also presented with potentially actionable Tier III & IV mutations. Patients with concurrent TP53 and RAS mutations had significantly reduced overall survival (6.1 months versus 21.1 months, p KRAS allele frequency, as defined by those with over 20% variant allele frequency (VAF), also demonstrated reduced overall survival (12.1 months versus 42.9 months, p = 0.04).ConclusionsIn addition to identifying patients with genomic alterations suitable for clinically proven standard of care therapeutic options, the 50 gene NGS panel has significant potential in identifying potentially actionable non-tier 1 mutations and therefore may become future standard clinical practice.</div

Distribution of concurrent mutations identified by next generation sequencing in colorectal cancer.

Individual patients with concurrent mutations in each column. Tiers 1–3 as outlined by the European Society of Medical Oncology Scale for Clinical Actionability of Molecular Targets.</p

Mutations identified using next generation sequencing as stratified by the European society of medical oncology scale for clinical actionability of molecular targets guidelines.

Mutations identified using next generation sequencing as stratified by the European society of medical oncology scale for clinical actionability of molecular targets guidelines.</p

ESMO tiers of mutational actionability.

BackgroundNext generation sequencing (NGS) is increasingly used in standard clinical practice to identify patients with potentially actionable mutations. Stratification of NGS mutation tiers is currently based on the European Society of Medical Oncology (ESMO) Scale for Clinical Actionability of Molecular Targets (ESCAT[E]) Tier I–V & X. Allele frequency is also increasingly recognised as an important prognostic tool in advanced cancer. The aim of this study was to determine the genomic mutations in metastatic colorectal cancer (CRC) in an Australian multicultural population and their influence on survival outcomes.MethodsNext generation sequencing with the 50-gene panel Oncomine Precision Assay™ was used on 180 CRC tissue samples obtained across six Sydney hospitals between June 2021 and March 2022.ResultsFrom 180 samples, 147 (82%) had at least one gene mutation identified with 68 (38%) having two or more concurrent mutations. Tier I variants included RAS wild-type [EI] in 73 (41%) and BRAF V600E [EIA] in 27 (15%). Non-tier I variants include 2 (1%) ERBB2 amplification [EIIB], 26 (15%) PIK3CA hotspot mutations [EIIIA] and 9 (5%) MET focal amplifications [EIIIA]. NGS testing revealed an additional 22% of cases with Tier II & III mutations. 43% of patients also presented with potentially actionable Tier III & IV mutations. Patients with concurrent TP53 and RAS mutations had significantly reduced overall survival (6.1 months versus 21.1 months, p KRAS allele frequency, as defined by those with over 20% variant allele frequency (VAF), also demonstrated reduced overall survival (12.1 months versus 42.9 months, p = 0.04).ConclusionsIn addition to identifying patients with genomic alterations suitable for clinically proven standard of care therapeutic options, the 50 gene NGS panel has significant potential in identifying potentially actionable non-tier 1 mutations and therefore may become future standard clinical practice.</div

S1 Data -

BackgroundNext generation sequencing (NGS) is increasingly used in standard clinical practice to identify patients with potentially actionable mutations. Stratification of NGS mutation tiers is currently based on the European Society of Medical Oncology (ESMO) Scale for Clinical Actionability of Molecular Targets (ESCAT[E]) Tier I–V & X. Allele frequency is also increasingly recognised as an important prognostic tool in advanced cancer. The aim of this study was to determine the genomic mutations in metastatic colorectal cancer (CRC) in an Australian multicultural population and their influence on survival outcomes.MethodsNext generation sequencing with the 50-gene panel Oncomine Precision Assay™ was used on 180 CRC tissue samples obtained across six Sydney hospitals between June 2021 and March 2022.ResultsFrom 180 samples, 147 (82%) had at least one gene mutation identified with 68 (38%) having two or more concurrent mutations. Tier I variants included RAS wild-type [EI] in 73 (41%) and BRAF V600E [EIA] in 27 (15%). Non-tier I variants include 2 (1%) ERBB2 amplification [EIIB], 26 (15%) PIK3CA hotspot mutations [EIIIA] and 9 (5%) MET focal amplifications [EIIIA]. NGS testing revealed an additional 22% of cases with Tier II & III mutations. 43% of patients also presented with potentially actionable Tier III & IV mutations. Patients with concurrent TP53 and RAS mutations had significantly reduced overall survival (6.1 months versus 21.1 months, p KRAS allele frequency, as defined by those with over 20% variant allele frequency (VAF), also demonstrated reduced overall survival (12.1 months versus 42.9 months, p = 0.04).ConclusionsIn addition to identifying patients with genomic alterations suitable for clinically proven standard of care therapeutic options, the 50 gene NGS panel has significant potential in identifying potentially actionable non-tier 1 mutations and therefore may become future standard clinical practice.</div

Overall survival of RAS-mutated metastatic colorectal cancer based on allele frequency.

Kaplan-Meier curve comparing KRAS high variant allele frequent as defined by over 20% (yellow) vs KRAS low variant allele frequent as defined by below 20% (blue). Dotted line represents median (12.1 months versus not reached, HR 2.2, 95%CI 1.3–6.7, p = 0.04).</p