Abstract 158P
Background
Single-cell RNA sequencing (scRNA-seq) has the ability to unveil uncommon cell populations. However, due to the high demand for tissue quality and cell viability, currently most scRNA-seq for pancreatic cancer was performed by surgical specimen or biopsy from metastatic sites. This study aims to establish a practical experience to help scientists perform primary pancreatic tumor scRNA-seq using EUS-FNB samples in real-world practice.
Methods
Two punctures from the same lesion using the same needle, without applying suction (Non-suction group) and with a negative pressure of 5 ml (Suction group) were evaluated. Single cell RNA sequencing libraries were prepared with Chromium Single cell 5’ Reagent Kits v2 (10X Genomics, USA) following the manufacturer’s protocol.
Results
A total of 20 patients were enrolled. The median age was 65.1 years old (range 46.6-83.2). Suction group achieved a cell preparation success rate of 80% (16/20) which was significantly higher than the 10% (2/20) success rate in the non-suction group (p<0.001). After the establishment of cell preparation protocol, we generate single-cell RNA transcriptomes for four patients, including two early stage (9,632 cells) and two late stage (4,592 cells). After quality control, 11,950 single cells were feasible for downstream analysis. Overall, 66% of cells (7,842) belonged to early stage and 34% (4,108) belonged to late stage. 12 major cell subtypes were identified across early and late stage. The proportion of cancer cells cluster-4 was significantly higher in late stage. Differentially expressed genes analysis showed UBE2C is the most highly expressed gene in cancer cells cluster-4. As external validation, in TCGA PAAD dataset, we found UBE2C high expression pancreatic cancer had significantly poor survival.
Conclusions
EUS-FNB with a negative pressure of 5 ml is feasible for single-cell sequencing in daily practice. A UBE2C high-expression subclone exists in early-stage pancreatic cancer and correlates with poor prognosis, potentially becoming a new therapeutic target in future studies.
Clinical trial identification
Protocol number: NCT05767697 Release date: 02 March, 2023.
Editorial acknowledgement
Legal entity responsible for the study
The authors.
Funding
National Health Research Institutes.
Disclosure
All authors have declared no conflicts of interest.
Resources from the same session
561P - Mechanisms of osimertinib resistance using circulating tumor DNA analyses for EGFR-mutated non-small cell lung cancer, results from ELUCIDATOR: A prospective observational multicenter study
Presenter: Daijiro Harada
Session: Poster Display
Resources:
Abstract
562P - First-line (1L) osimertinib (osi) ± platinum-pemetrexed in patients (pts) with EGFRm advanced NSCLC: FLAURA2 China cohort
Presenter: Yan Yu
Session: Poster Display
Resources:
Abstract
563P - Real-world effectiveness and safety of first-line osimertinib for EGFR-mutated advanced NSCLC in China (FLOURISH study)
Presenter: Jianya Zhou
Session: Poster Display
Resources:
Abstract
564P - Co-occurring EGFR p.E709X mutation affects the treatment response to the third-generation EGFR-TKIs in EGFR p.G719X-mutant patients with advanced NSCLC
Presenter: Wen Feng Fang
Session: Poster Display
Resources:
Abstract
565P - Genome-guided targeted therapy combination improves survival in patients with advanced EGFR mutation positive NSCLC failing osimertinib
Presenter: Molly Li
Session: Poster Display
Resources:
Abstract
566P - Safety of tepotinib + osimertinib in EGFR-mutant NSCLC with MET amplification after first-line osimertinib
Presenter: Chong Kin Liam
Session: Poster Display
Resources:
Abstract
567P - Furmonertinib in combination with bevacizumab and intrathecal chemotherapy as later-line re-challenge treatment in EGFR –mutated NSCLC patients with leptomeningeal metastasis after third-generation EGFR-TKIs treatment failure
Presenter: Fang Cun
Session: Poster Display
Resources:
Abstract
568P - First-line (1L) osimertinib + platinum-pemetrexed in EGFR-mutated (EGFRm) advanced NSCLC: Updated FLAURA2 safety run-in (SRI) results
Presenter: David Planchard
Session: Poster Display
Resources:
Abstract
569P - Whole-transcriptome sequencing of transformed small-cell lung cancer from EGFR-mutated lung adenocarcinoma reveals LUAD–like and SCLC–like subsets
Presenter: Chan-Yuan Zhang
Session: Poster Display
Resources:
Abstract
570P - First-line osimertinib for patients with advanced NSCLC harboring EGFR mutations: A real-world study
Presenter: Wenxiang Ji
Session: Poster Display
Resources:
Abstract