Abstract
PurposePathologic staging of bladder cancer patients remains a challenge. Standard-of-care histology exhibits limited sensitivity in detection of micrometastases, which can increase risk of cancer progression and delay potential adjuvant therapies. Here, we sought to develop a proof of concept novel molecular approach to improve detection of cancer micrometastasis.Experimental DesignWe combined fluorescence activated cell sorting and next-generation sequencing and performed whole-exome sequencing of total cancer cells and cancer cell subpopulations in multiple tumor specimens and regional lymph nodes in a single patient with muscle-invasive urothelial carcinoma of the bladder following radical cystectomy.ResultsMean allele frequency analysis demonstrated a significant correlation between primary tumor cancer cells and cancer cells isolated from the lymph nodes, confirming lymph node disease despite negative pathologic staging. RNA-sequencing revealed intratumoral heterogeneity as well as enrichment for immune system and lipid metabolism gene sets in the micrometastatic cancer cell subpopulations.ConclusionsOur analysis illustrates how next-generation sequencing of cancer cell subpopulations may be utilized to enrich for cancer cell markers and enhance detection of bladder cancer micrometastases to improve pathologic staging and provide insight into cancer cell biology.
Highlights
Latent recurrences despite favorable pathology has been a longstanding conundrum in surgical oncology
Experimental Design: We combined fluorescence activated cell sorting and nextgeneration sequencing and performed whole-exome sequencing of total cancer cells and cancer cell subpopulations in multiple tumor specimens and regional lymph nodes in a single patient with muscle-invasive urothelial carcinoma of the bladder following radical cystectomy
As bladder cancer progresses based on a predictable pattern of lymphatogenous metastasis with recurrence attributed to lymph node micrometastases, improved pathologic staging www.impactjournals.com/oncotarget of lymph nodes may add precision to selecting adjuvant therapy as well as stratifying patients with metastatic disease [4]
Summary
Latent recurrences despite favorable pathology has been a longstanding conundrum in surgical oncology. In patients with muscle-invasive urothelial carcinoma of the bladder, radical cystectomy and pelvic lymph node dissection provides local surgical control. As bladder cancer progresses based on a predictable pattern of lymphatogenous metastasis with recurrence attributed to lymph node micrometastases, improved pathologic staging www.impactjournals.com/oncotarget of lymph nodes may add precision to selecting adjuvant therapy as well as stratifying patients with metastatic disease [4]. Cancer stem cells (CSC) have been implicated as a source of early metastasis and disease progression [5] They represent a self-renewing and pluripotent population that can reconstitute all cell types of an individual tumor. We prospectively purified and characterized total and EPCAM+CD44+CD49f+ subpopulations in fresh, surgicallyresected tissue from multiple regions of a bladder tumor and regional lymph nodes by whole-exome sequencing and RNA-sequencing in a single patient with clinically localized muscle-invasive urothelial bladder carcinoma
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