Clinical Sensitivity Verification Study of Circulating Tumor Cells Gene Mutation Detection From Advanced NSCLC Patients

Abstract Objectives To analyze the clinical relevance of heterogeneous phenotypes of peripheral circulating tumor cells (CTCs) in non-small cell lung cancer (NSCLC). Materials and Methods CTCs in 5 mL venous blood were enriched using the Canpatrol™ CTC technique in 82 NSCLC patients. And then, CTCs were subjected to RNA in situ hybridization with a combination of epithelial (EpCAM and CK8/18/19) and mesenchymal (vimentin and TWIST1) markers. Results According to the fluorescent dots, CTCs were classified into three groups, including epithelial CTCs (E-CTC), hybrid epithelial/mesenchymal phenotypes (E/M-CTCs) and mesenchymal CTCs (M-CTCs). In 82 NSCLC cohort, only 2 patients didn’t detect CTCs, the overall CTCs detection rate was 97.5% (80/82). For 60 treatment naïve NSCLC, only one patient didn’t detect CTCs. The median number of total CTCs, hybrid E/M phenotype CTCs, E-CTCs and M-CTCs per 5 mL blood was 22 (range 1–90), 13 (range 0–83), 1 (range 0–17 and 0–47), respectively. Hybrid E/M CTCs, especially the e = m-CTCs, significantly differed between patients with and without distant metastasis. M-CTCs in advanced NSCLC patients were significantly more than the numbers observed in early stage patients. Patients with pure hybrid E/M-CTCs showed a lower proportion in distant metastasis positive cohort compared to negative ones (7% vs 22%), while patients with E + E/M CTCs (20% vs 9%) and E/M + M CTCs (33% vs 20%) showed a higher proportion. CTCs dynamic changes after treatment in 12 advanced NSCLC patients suggested that hybrid E/M-CTCs were related to the primary tumor size at baseline, while M-CTCs may suggest the progression of NSCLC. Conclusion We concluded that E-CTCs with a hybrid E/M phenotype are associated to metastasis in therapy-naïve NSCLC patients.

Download Full-text

Longitudinal Change of Circulating Tumor Cells During Chemoradiation Therapy and Its Correlation with Prognosis in Advanced Non-Small Cell Lung Cancer Patients

10.21203/rs.3.rs-37672/v1 ◽

2020 ◽

Author(s):

Jun Liu ◽

Yongping Liu ◽

Cheng Gu ◽

Lei Zhang ◽

Xujing Lu

Keyword(s):

Lung Cancer ◽

Small Cell Lung Cancer ◽

Treatment Response ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Cell Lung Cancer ◽

Advanced Nsclc ◽

Small Cell ◽

Small Cell Lung ◽

Nsclc Patients

Abstract Background: This study aimed to investigate the association of circulating tumor cells (CTCs) change during chemoradiation with the treatment response and survival profiles in advanced non-small cell lung cancer (NSCLC) patients.Methods: 58 advanced NSCLC patients underwent concurrent chemoradiation were enrolled, then their peripheral blood samples were collected pre-chemoradiation and at 1 months post-chemoradiation to assess the CTCs using a CTC-Biopsy system. Moreover, CTCs were classified as CTCs positive and CTCs negative according to CTCs’ count, and CTCs’ change was calculated. Additionally, response of chemoradiation was evaluated at 1 months post-chemoradiation, then progression-free survival (PFS) and overall survival (OS) were assessed. Results: Pre-chemoradiation CTCs positive was associated with increased TNM stage, but not correlated with other clinicopathologic characteristics. After chemoradiation, the CTCs’ number (1.0 (0.0-3.0) vs. 4.0 (2.0-10.0)) and the percentage of CTCs positive cases (37.9% vs. 77.6%) were both decreased compared with those prior to chemoradiation. Regarding treatment response, pre-chemoradiation CTCs positive was associated with lower partial response; post-chemoradiation CTCs positive was associated with reduced disease control rate; while CTCs’ change during chemoradiation was not associated with treatment response. Kaplan–Meier curves showed that post-chemoradiation CTCs positive and increased CTCs’ number during chemoradiation were associated with reduced PFS, then multivariate Cox’s regression analysis disclosed that they independently predicted decreased PFS. However, no correlation of CTCs status or CTCs’ change with OS was observed. Conclusions: Longitudinal monitoring of CTCs may provide important reflection for the prognosis in chemoradiation treated advanced NSCLC patients.

Download Full-text

Abstract 1332: High Throughput isolation and expansion of circulating tumor cells (CTCs) from Non-small cell lung cancer (NSCLC) patients for personalized treatments

10.1158/1538-7445.am2019-1332 ◽

2019 ◽

Author(s):

Mina Zeinali ◽

Maggie Lee ◽

Arthi Nadhan ◽

Anvya Mathur ◽

Wei Huang ◽

...

Keyword(s):

Lung Cancer ◽

Small Cell Lung Cancer ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

High Throughput ◽

Cell Lung Cancer ◽

Small Cell ◽

Small Cell Lung ◽

Nsclc Patients

Download Full-text

Abstract 775: Longitudinal investigation of PD-L1 expression on circulating tumor cells (CTCs) in non-small cell lung cancer (NSCLC) patients treated with nivolumab

10.1158/1538-7445.am2020-775 ◽

2020 ◽

Author(s):

Yasuhiro Koh ◽

Mio Ikeda ◽

Shunsuke Teraoka ◽

Koichi Sato ◽

Nahomi Tokudome ◽

...

Keyword(s):

Lung Cancer ◽

Small Cell Lung Cancer ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Cell Lung Cancer ◽

Small Cell ◽

Small Cell Lung ◽

Longitudinal Investigation ◽

Nsclc Patients

Download Full-text

Determination of PD-L1 Expression in Circulating Tumor Cells of NSCLC Patients and Correlation with Response to PD-1/PD-L1 Inhibitors

Cancers ◽

10.3390/cancers11060835 ◽

2019 ◽

Vol 11 (6) ◽

pp. 835 ◽

Cited By ~ 28

Author(s):

Melanie Janning ◽

Franca Kobus ◽

Anna Babayan ◽

Harriet Wikman ◽

Janna-Lisa Velthaus ◽

...

Keyword(s):

Tumor Cells ◽

Circulating Tumor Cells ◽

Clinical Utility ◽

Small Cell Lung ◽

Cellsearch System ◽

Independent System ◽

Nsclc Patients ◽

System 1 ◽

Key Questions

Circulating tumor cells (CTCs) hold great potential to answer key questions of how non-small cell lung cancer (NSCLC) evolves and develops resistance upon anti-PD-1/PD-L1 treatment. Currently, their clinical utility in NSCLC is compromised by a low detection rate with the established, Food and Drug Administration (FDA)-approved, EpCAM-based CellSearch® System. We tested an epitope-independent method (ParsortixTM system) and utilized it to assess PD-L1 expression of CTCs from NSCLC patients. We prospectively collected 127 samples, 97 of which were analyzed with the epitope-independent system in comparison to the CellSearch system. CTCs were determined by immunocytochemistry as intact, nucleated, CD45−, pankeratins (K)+ cells. PD-L1 status of CTCs was evaluated from 89 samples. With the epitope-independent system, ≥1 CTC per blood sample was detected in 59 samples (61%) compared to 31 samples (32%) with the EpCAM-based system. Upon PD-L1 staining, 47% of patients harbored only PD-L1+CTCs, 47% had PD-L1+ and PD-L1−CTCs, and only 7% displayed exclusively PD-L1−CTCs. The percentage of PD-L1+CTCs did not correlate with the percentage of PD-L1+ in biopsies determined by immunohistochemistry (p = 0.179). Upon disease progression, all patients showed an increase in PD-L1+CTCs, while no change or a decrease in PD-L1+CTCs was observed in responding patients (n = 11; p = 0.001). Our data show a considerable heterogeneity in the PD-L1 status of CTCs from NSCLC patients. An increase of PD-L1+CTCs holds potential to predict resistance to PD-1/PD-L1 inhibitors.

Download Full-text

Antibody-independent ApoStream technology isolates folate receptor alpha (FRA)–positive circulating tumor cells from blood of non-small cell lung adenocarcinoma patients.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.15_suppl.e21028 ◽

2012 ◽

Vol 30 (15_suppl) ◽

pp. e21028-e21028

Author(s):

Darren W. Davis ◽

Christopher Neal ◽

Vishal Gupta ◽

Vladislava O. Melnikova ◽

Jacky Woo ◽

...

Keyword(s):

Lung Adenocarcinoma ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Folate Receptor ◽

Small Cell ◽

Normal Donor ◽

Small Cell Lung ◽

Folate Receptor Alpha ◽

Receptor Alpha ◽

Nsclc Patients

e21028 Background: The ability to enrich and interrogate circulating tumor cells (CTCs) from blood may allow for the analysis of metastatic dissemination and potential use of CTCs as surrogates for monitoring drug efficacy. We developed ApoStream, a dieletrophoresis field-flow fractionation based platform, for antibody-independent CTC isolation. We demonstrated that ApoStream successfully isolates non-small cell lung adenocarcinoma (NSCLC) CTCs that express Folate Receptor alpha (FRA), a GPI-anchored receptor, which has emerged as a cancer biomarker and potential therapeutic target in multiple cancer types. Methods: ApoStream technology was used to enrich CTCs from NSCLC patients’ blood. CTC enrichment by ApoStream was compared to that of the FDA cleared CellSearch CTC kit. A multiplexed immunofluorescent assay was developed to enable CTC enumeration (Cytokeratin+/CD45-/DAPI+ cells) and analysis of FRA expression (detection by murine antibody clone 26B3) using single cell quantitative laser scanning cytometry (LSC). Results: In a side-by-side comparison with the CellSearch CTC kit, ApoStream isolated significantly higher numbers of CTCs in 9 metastatic adenocarcinoma NSCLC patients (Apostream: mean =139, range 3-487 per 7.5 mL of blood, versus CellSearch kit: mean =2, range 0-8 per 7.5 mL of blood, n= 9, p=0.041). All patients were found to be CTC-positive by ApoStream, while only 3 of 9 (33%) patients were CTC-positive based on the CellSearch kit. LSC analysis demonstrated that 8-33% of all CTCs isolated expressed FRA and that FRA expression was confined to CTCs only. No false positive CTCs and no FRA-expressing cells were isolated by ApoStream from normal donor blood (n=15). Conclusions: All NSCLC adenocarcinoma patients analyzed had FRA-positive CTCs, suggesting that FRA may play a key role in metastasis and that screening of patients with the ApoStream CTC isolation system may identify patients who could benefit from FRA-targeted therapy.

Download Full-text