Molecular analysis of archival diagnostic prostate cancer biopsies identifies genomic similarities in cases with progression post-radiotherapy, and those with de novo metastatic disease.

Charlton, P.; O'Reilly, D.; Philippou, Y.; Rao, S. R.; Lamb, A. D.; Mills, I. G.; Higgins, G.; Hamdy, F. C.; Verrill, C.; Buffa, F. M.; Bryant, R. J.

doi:10.1101/2023.09.04.555868

bioRxiv · 10.1101/2023.09.04.555868

Molecular analysis of archival diagnostic prostate cancer biopsies identifies genomic similarities in cases with progression post-radiotherapy, and those with de novo metastatic disease.

Charlton, P.·O'Reilly, D.·Philippou, Y.·Rao, S. R.·Lamb, A. D.·Mills, I. G.·Higgins, G.·Hamdy, F. C.·Verrill, C.·Buffa, F. M.·Bryant, R. J.

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Abstract

PurposeIt is important to identify molecular features that improve prostate cancer (PCa) risk stratification before radical treatment with curative intent. Molecular analysis of historical diagnostic formalin-fixed paraffin-embedded (FFPE) prostate biopsies from cohorts with post-radiotherapy (RT) long-term clinical follow-up has been limited. Utilizing parallel sequencing modalities, we performed a proof-of-principle sequencing analysis of historical diagnostic FFPE prostate biopsies. We compared patients with i) stable PCa post-primary or salvage RT (sPCa), ii) progressing PCa post-RT (pPCa), and iii) de novo metastatic PCa (mPCa). Experimental DesignA cohort of 19 patients with diagnostic prostate biopsies (n=6 sPCa, n=5 pPCa, n=8 mPCa) and mean 4 years 10 months follow-up (diagnosed 2009-2016) underwent nucleic acid extraction from demarcated malignancy. Samples underwent 3RNA sequencing (3RNAseq) (n=19), nanoString analysis (n=12) and Illumina 850k methylation (n=8) sequencing. Bioinformatic analysis was performed to coherently identify differentially expressed genes (DEGs) and methylated genomic regions (MGRs). Results18 of 19 samples provided useable 3RNAseq data. Principal Component Analysis (PCA) demonstrated similar expression profiles between pPCa and mPCa cases, versus sPCa. Coherently differentially methylated probes between these groups identified [~]600 differentially MGRs. The top 50 genes with increased expression in pPCa patients were associated with reduced progression-free survival post-RT (p<0.0001) in an external cohort. Conclusions3RNAseq, nanoString and 850K-methylation analyses are each achievable from historical FFPE diagnostic pre-treatment prostate biopsies, unlocking the potential to utilize large cohorts of historic clinical samples. Profiling similarities between individuals with pPCa and mPCa suggests biological similarities and historical radiological staging limitations, which warrant further investigation.

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Charlton, P., O'Reilly, D., Philippou, Y., Rao, S. R., Lamb, A. D., Mills, I. G., Higgins, G., Hamdy, F. C., Verrill, C., Buffa, F. M., Bryant, R. J.. 2023-09-06. Molecular analysis of archival diagnostic prostate cancer biopsies identifies genomic similarities in cases with progression post-radiotherapy, and those with de novo metastatic disease.. https://doi.org/10.1101/2023.09.04.555868

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