The use of cfDNA quantification to predict adenocarcinoma at an early stage in high-risk (aged >50 years and FOBT positive) subjects seems to be promising but needs more sensitive methods to improve cfDNA detection.
FOLFIRINOX, a combination of chemotherapy drugs (Fluorouracil, Oxaliplatin, Irinotecan -FOI), provides the best clinical benefit in pancreatic ductal adenocarcinoma (PDAC) patients. In this study we explore the role of miRNAs (MIR) as modulators of chemosensitivity to identify potential biomarkers of response. We find that 41 and 84 microRNA inhibitors enhance the sensitivity of Capan1 and MiaPaCa2 PDAC cells respectively. These include a MIR1307-inhibitor that we validate in further PDAC cell lines. Chemotherapy-induced apoptosis and DNA damage accumulation are higher in MIR1307 knock-out (MIR1307KO) versus control PDAC cells, while re-expression of MIR1307 in MIR1307KO cells rescues these effects. We identify binding of MIR1307 to CLIC5 mRNA through covalent ligation of endogenous Argonaute-bound RNAs cross-linking immunoprecipitation assay. We validate these findings in an in vivo model with MIR1307 disruption. In a pilot cohort of PDAC patients undergoing FOLFIRONX chemotherapy, circulating MIR1307 correlates with clinical outcome.
Circulating tumour DNAs and non-coding RNAs present in body fluids have been under investigation as tools for cancer diagnosis, disease monitoring, and prognosis for many years. These so-called liquid biopsies offer the opportunity to obtain information about the molecular make-up of a cancer in a minimal invasive way and offer the possibility to implement theranostics for precision oncology. Furthermore, liquid biopsies could overcome the limitations of tissue biopsies in capturing the complexity of tumour heterogeneity within the primary cancer and among different metastatic sites. Liquid biopsies may also be implemented to detect early tumour formation or to monitor cancer relapse of response to therapy with greater sensitivity compared with the currently available protein-based blood biomarkers. Most colorectal cancers are often diagnosed at late stages and have a high mortality rate. Hence, biomolecules as nucleic acids present in liquid biopsies might have prognostic potential and could serve as predictive biomarkers for chemotherapeutic regimens. This review will focus on the role of circulating tumour DNAs and non-coding RNAs as diagnostic, prognostic, and predictive biomarkers in the context of colorectal cancer.
Trastuzumab in combination with chemotherapy represents the standard of care in HER2-positive advanced gastro-esphageal cancer (GOC), but development of resistance limits response. MicroRNAs (miRs) modulate key pathways in GOC. Identification of miRs responsible for resistance to HER2 inhibition may help stratify patients, predict response and define novel strategies to restore drug sensitivity. A high-throughput LNA™ miR-inhibitor screen in HER2-positive NCI-N87 and HER2-negative FLO-1 GOC cell lines was performed in order to identify potential miRs involved in trastuzumab sensitivity/resistance. Cells were treated with a combination of cisplatin, 5FU, and trastuzumab. MiRs were considered significant if LNAs caused >40% reduction in cell viability 72 hours post treatment relative to controls with a t-test p-value of <0.001 across 3 biological replicates. Putative hits with higher baseline miR levels on nCounter® NCI-N87 cell line analysis were validated in two HER2-positive cell lines by calculating the difference (Δ) in cell viability between cells treated with trastuzumab+chemotherapy versus ‘no drug' control. MiR levels in translational samples from ten HER2-positive GOC patients recruited to the FOrMAT trial (NCT02112357) were analysed by ddPCR. MiR levels were categorized as ‘low' or ‘high' relative to median expression and correlated with clinical response. Kaplan-Meier curves for PFS and OS were generated. In NCI-N87 and FLO-1 cell lines, 59 and 37 LNAs respectively caused >40% reduction in cell viability compared to controls (p<0.001). Eighteen miRs were significant exclusively in the NCI-N87 cell line screen, suggesting potential involvement in the HER2-signaling pathway. nCounter® analysis showed 6 of these were elevated at baseline in NCI-N87: miR-1260, miR-148-3p, miR-28-3p, miR-331-3p, miR-7-5p and miR-365. Inhibition of miR-148a-3p increased sensitivity to trastuzumab + 5FU+cisplatin in both the NCI-N87 cell line (Δ 67% versus Δ 55% in the control) and OACP4C cell line (Δ 50% versus Δ 47% in the control). Patients who responded to trastuzumab + chemotherapy in FOrMAT had a trend towards lower average miR-148a-3p plasma levels than those who failed to respond. Median PFS for plasma miR-148a-3p ‘low' subgroup was 146 days versus 154 days in ‘high' subgroup. Median OS for plasma miR-148a-3p ‘low' subgroup was 357 days versus 226 days for miR-148a-3p ‘high' subgroup. Our screen identified a panel of miRs associated with GOC resistance to HER2 inhibitors in combination with chemotherapy. Inhibition of these miRs affects GOC cell viability and restores sensitivity. MiR-148a-3p emerged as a potential biomarker of resistance and further evaluation is underway in the PLATFORM trial (NCT02678182). Citation Format: Hazel Lote, Andrea Lampis, George Vlachogiannis, Jens Hahne, Sing-Yu Moorcraft, Michael Davidson, Carolin Fong, Ruwaida Begum, Matteo Fassan, Sheela Rao, David Watkins, Naureen Starling, Ian Chau, David Cunningham, Nicola Valeri. MicroRNAs as biomarkers of resistance to HER2 inhibition in combination with chemotherapy in gastro-esophageal cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 258.
ntroduction: FOLFIRINOX-regime is a combination-chemotherapy that provides the best clinical benefit in pancreatic cancer (PC) patients, but is associated with severe toxicity. Aim of this study is to explore the role of miRNAs (MIR) as modulators of chemosensitivity and their potential as biomarkers of sensitivity to FOLFIRINOX-chemotherapy. Methods: High-throughput-screening (HTS) of 997 LNA-MIR-inhibitors was performed in PC cell lines (Capan1, MiaPaCa2) treated with a combination of Fluorouracil (F), Oxaliplatin (O) and Irinotecan (I) that mimics FOLFIRINOX-regime. Cell viability was monitored by CellTiter-Blue assay. Validation experiments were carried out with miRvana probes. MIR expression was assessed by TaqMan-assay. Apoptosis was measured by Flow-cytometry and western-blotting. Knock-out of microRNA was acheived by CRISPR-CAS9 in MiaPaca2 cells (MIR1307KO). Results: 41 and 84 miRNA-inhibitors enhanced FOI activity by >30% (p<0.001) in Capan1 and MiaPaCa2. These included MIR1307-inhibitor that was validated in Capan1, MiaPaCa2, Panc1, AspC1, BxPC3, and Su86.86 cell lines. The proportion of cells killed by FOI in comparison to DMSO was greater in cells transfected with MIR1307 inhibitor when compared to NEG CTRL, making the effect of this MIR specific for chemotherapy. MIR1307 was over-expressed in tumour compared to matched-adjacent tissue in 40/60 human PC cases, confirming clinical relevance. MIR1307KO cells were more sensitive to FOI than WT cells. Chemotherapy-induced apoptosis was higher in MIR1307KO cells (caspase 3/7 activity and annexin-V positivity). We observed significant upregulation of different markers of DNA damage (pH2AX2, 8OHdG, DNA breaks in Comet assay) in MIR1307KO cells treated with FOI in comparison to WT treated cells. Re-expression of MIR1307 in MIR1307KO cells could increase resistance to FOI chemotherapy and protected from FOI-induced DNA damage. Bioinformatics analysis identified MIR1307 binding-sites within a number of genes involved in the DNA-repair pathway (p<0.001, folding energy value greater than -12 Kcal/mol). Conclusions: We identified miR-1307 as a potential modulator of sensitivity to FOI-chemotherapy in PC. We showed that miR-1307 inhibition impairs the ability of PC cells to recover from chemotherapy damage and therefore enhances its activity. The assessemnt of it potential as predictive biomarker of response in PC patients is ongoing. Citation Format: Pietro Carotenuto, Domenico Zito, Maria C. Previdi, Maya Raj, Matteo Fassan, Andrea Lampis, Francesco Scalafani, Andrea Lanese, Ian Said-Huntingford, Jens C. Hahne, Kate Young, Ruwaida Begum, Zakaria Ethiar, Andrew Wotherspoon, Naureen Starling, Anguraj Sadanandam, David Cunningham, Ian Chau, Paul Workman, Rajesh Chopra, Nicola Valeri, Chiara Braconi. MIR1307 mediates pancreatic cancer resistance to FOLFIRINOX chemotherapy by affecting response to DNA damage [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4977.
Background: AE-mABs (cetuximab and/or panitumumab) have been approved for the treatment of RAS wild-type (WT) metastatic colorectal cancer (mCRC) patients (pts). Indeed, previous studies have identified mutations (MTs)/amplifications in RAS/RAF/MEK kinase pathway as the main genetic events promoting primary and acquired resistance to these mABs. RAS status is frequently established on archival material, as tumour re-biopsy may not be always feasible. PROSPECT-C is a prospective trial aiming to define novel and known mechanisms of resistance to AE-mABs by obtaining repeated biopsies and sequential bloods from pts receiving AE-mABs for chemo-refractory mCRC. Here we present the preliminary results of the PROSPECT-C liquid biopsy study where the goals were to: 1) confirm the ability of ctDNA to determine RAS status prior to treatment; 2) determine any discordance of RAS status between archival material and ctDNA; 3) confirm the ability of ctDNA to track emerging MTs in the RAS pathway. Materials and Methods: Plasma was collected at baseline (BL), every 4 weeks and at progression (PD). ctDNA was isolated using the QIAamp Circulating Nucleic Acid Kit (Qiagen) and analyzed by digital droplet PCR (QX200 Bio-Rad) for MTs in KRAS-G12D, KRAS-G13D, KRAS-G12V, KRAS-Q61HA>T, KRAS-Q61HA>C, and BRAF-V600E. In the next steps, plasma samples with no MTs in the first series of hotspots will be tested for the remaining KRAS hotspots, NRAS, EGFR, PIK3CA MTs and KRAS, c-MET and HER-2 amplifications (Data to be presented). Results: Twenty-four pts (all treated with AE-mAB monotherapy) with KRAS WT tumors (from archival tissue) have been treated on this ongoing study; 16/24 had ctDNA analysis (mean age 62 years, 62.5% males) so far. All pts were heavily pre-treated; 58.3%, 31.3% and 12.5% received 2, 3 and 4 lines of prior therapies respectively. Of 15 pts with BL samples, 5 had MTs at BL; 2 with KRAS-G12D, 1 with KRAS Q61HA>T and 2 pts with BRAF-V600E (in the latter case concordance was found with the archival material). Best response was found to be PD in 4/5 pts and stable disease in 1/5pt with BL MTs; 2 with BRAF MTs progressed in <2 months (mo). The median progression free survival (PFS) for pts with BL MTs was 1.8 mo. Five pts with no MT at BL developed one or more MTs later; those included 4 with KRAS-G12D, 3 with KRAS-G13D and 2 with KRASQ61HA>T (median PFS = 4.8 mo). 3/15 (20%) pts showed discordance in RAS MTs between archival material (WT) and baseline bloods (MT); all of them progressed within 3 mo. Conclusions: Liquid biopsies can be a useful tool in determining and tracking RAS MTs in pts undergoing anti-EGFR therapy for mCRC. ctDNA analysis to assess RAS mutational status prior to receiving AE-mABs in our series showed 20% discordance between the archival solid and BL liquid biopsies, which may account for resistance to these therapies. Citation Format: Khurum Khan, George Vlachogianis, David Cunningham, Jens Hahne, Mahnaz Darvish-Damavandi, Sarah Barton, Francesco Trevisani, Giulia Mentrast, Clare Peckitt, Andrea Lampis, Chiara Braconi, Nasir Khan, Ruwaida Begum, Naureen Starling, Sheela Rao, David Watkins, Annette Bryant, Ian Chau, Nicola Valeri. Validation of the role of circulating tumor DNA (ctDNA) in tracking mechanisms of resistance to anti-EGFR monoclonal antibodies (AE-mABs): preliminary results of the PROSPECT-C prospective trial. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3589. doi:10.1158/1538-7445.AM2015-3589
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.