Abstract 13P
Background
HER2DX is a genomic test for early HER2+ breast cancer clinically available since January 2022 as a Laboratory Developed Test. Based on the expression of 27 genes (nCounter) plus tumor and nodal staging, it predicts the risk of relapse (risk-score), the probability of achieving a pathological complete response (pCR-score), and the individual levels of ERBB2 expression (ERBB2-score). Here, we report the analytical validation of HER2DX performed in a central laboratory (CLab) having as a reference the development lab (DLab).
Methods
Intra- and inter- labs HER2DX scores were compared by testing a set of FFPE samples previously analyzed at DLab. All scores ranged from 0 to 100 and pre-defined cut-offs were used to get HER2DX-groups. Repeatability and reproducibility were evaluated from different tissue sections, RNA, or FFPE blocks. Simulations (n=1·106) were used to calculate diagnostic values (sensitivity, specificity, positive and negative predictive values, and accuracy). Robustness was measured by evaluating the interference of non-tumor tissue and by using different RNA quantities. Differences due to the nCounter instrument, Tagset lot, and Tagset defrost cycles were also evaluated. HER2DX performance using a RNAseq platform (Illumina Exome Panel) was compared to nCounter.
Results
Repeatability analysis within CLab in 10 samples showed a maximal standard error among the 3 scores of 0.94 (scale 0-100). Reproducibility starting from RNA was analyzed in 20 samples, and the probabilities of +/- 5 units difference in the risk-score, pCR-score, and ERBB2-scores were 0.5%, 5.2%, and 0.2%, respectively. Starting from FFPE blocks, correlation of scores between labs were >0.97. In the simulation, all diagnostic values were >90% for the three scores, showing a high association in HER2DX-group classification between labs. Robustness at low tumor content samples (10%) or low RNA quantity (100 ng) was acceptable for the assay. No significant differences were observed across different nCounter instruments, Tagset lots and defrost cycles at CLab. Data from RNAseq and nCounter platforms were highly correlated in 30 analyzed RNA samples.
Conclusions
Analytical validation of HER2DX has proven it to be suitable for its intended purpose.
Legal entity responsible for the study
Reveal Genomics, S.L.
Funding
Reveal Genomics, S.L.
Disclosure
M. Marín-Aguilera: Financial Interests, Institutional, Full or part-time Employment, Product Manager: Reveal Genomics, S.L. G. Villacampa: Financial Interests, Personal, Invited Speaker, Invited speaker in a course: MSD; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Invited Speaker, Invited speaker in an internal training: Pierre Fabre, GSK; Financial Interests, Personal, Invited Speaker, Internal discussion about the interpretation of some published results: Pfizer; Financial Interests, Personal, Other, Collaborations with specific projects: Reveal Genomics. F. Brasó-Maristany: Financial Interests, Personal, Invited Speaker: Reveal Genomics. J.A. Puig-Butille: Financial Interests, Personal, Invited Speaker: Thermo Fisher; Financial Interests, Personal, Funding: CareDX. A. Vivancos: Financial Interests, Personal, Advisory Board: Roche, Bristol Meyers Squibb, Guardant Health, Bayer, Incyte; Financial Interests, Personal, Stocks/Shares: Reveal Genomics; Financial Interests, Institutional, Research Grant, Preclinical Research Grant: Bristol Meyers Squibb, Roche, Incyte. J. Matito: Financial Interests, Personal, Full or part-time Employment: Reveal Genomics. C.M. Perou: Financial Interests, Personal, Invited Speaker: GeneCentric Therapeutics; Financial Interests, Personal, Stocks/Shares: GeneCentric Therapeutics, Bioclassifier LLC, Reveal Genomics; Financial Interests, Personal, Royalties, from PAM50 licensing: Bioclassifier LLC; Financial Interests, Personal, Royalties: GeneCentric Therapeutics; Non-Financial Interests, Advisory Role: GeneCentric Therapeutics, Bioclassifier LLC, Reveal Genomics. P. Villagrasa Gonzalez: Financial Interests, Personal and Institutional, Member of the Board of Directors: Reveal Genomics; Financial Interests, Personal and Institutional, Proprietary Information: Reveal Genomics. A. Prat: Financial Interests, Personal, Invited Speaker: Roche; Financial Interests, Personal, Invited Speaker, Lecture fees: Novartis, Daiichi Sankyo; Financial Interests, Personal, Advisory Board, Advisory role/consultancy: Novartis, Pfizer, Oncolytics Biotech, Guardant Health, Peptomyc; Financial Interests, Institutional, Invited Speaker, Clinical trials: Daiichi Sankyo; Financial Interests, Institutional, Other, Contracted research: Boehringer, Medica Scientia inno. Research; Financial Interests, Personal, Advisory Board: AstraZeneca; Financial Interests, Personal, Invited Speaker, Leadership role: Reveal Genomics, SL.; Financial Interests, Personal, Stocks/Shares: Reveal Genomics, Oncolytics Biotech; Financial Interests, Personal, Royalties: Reveal Genomics; Financial Interests, Institutional, Invited Speaker: Roche, AstraZeneca, Novartis; Financial Interests, Personal and Institutional, Invited Speaker: Daiichi Sankyo; Non-Financial Interests, Institutional, Other, Leadership roles: Patronage committee: SOLTI Foundation, Actitud Frente al Cáncer Foundation; Non-Financial Interests, Personal, Other, Asociación Española de Investigación sobre el Cáncer: ASEICA. J. Parker: Financial Interests, Personal, Other, Advisory: Reveal Genomics, Bristol Myers Squibb; Financial Interests, Personal, Full or part-time Employment: LifeEdit Therapeutics; Financial Interests, Personal, Ownership Interest: Reveal Genomics, GeneCentric; Financial Interests, Personal, Royalties: Veracyte. L. Pare Brunet: Financial Interests, Institutional, Full or part-time Employment: Reveal Genomics S.L.; Financial Interests, Institutional, Other, Patent pending: In vitro method for the prognosis of patients suffering from HER2-positive breast cancer - 905333: Reveal Genomics S.L. All other authors have declared no conflicts of interest.