Clinical relevance of the combined analysis of circulating tumor cells and anti-tumor T-cell immunity in metastatic breast cancer patients

Background: Metastatic breast cancer (mBC) is a heterogeneous disease with varying responses to treatments and clinical outcomes, still requiring the identification of reliable predictive biomarkers. In this context, liquid biopsy has emerged as a powerful tool to assess in real-time the evolving landscape of cancer, which is both orchestrated by the metastatic process and immune-surveillance mechanisms. Thus, we investigated circulating tumor cells (CTCs) coupled with peripheral T-cell immunity to uncover their potential clinical relevance in mBC. Methods: A cohort of 20 mBC patients was evaluated, before and one month after starting therapy, through the following liquid biopsy approaches: CTCs enumerated by a metabolism-based assay, T-cell responses against tumor-associated antigens (TAA) characterized by interferon-γ enzyme-linked immunosorbent spot (ELISpot), and the T-cell receptor (TCR) repertoire investigated by a targeted next-generation sequencing technique. TCR repertoire features were characterized by the Morisita’s overlap and the Productive Simpson Clonality indexes, and the TCR richness. Differences between groups were calculated by Fisher’s, Mann-Whitney or Kruskal-Wallis test, as appropriate. Prognostic data analysis was estimated by Kaplan-Meier method. Results: Stratifying patients for their prognostic level of 6 CTCs before therapy, TAA specific T-cell responses were detected only in patients with a low CTC level. By analyzing the TCR repertoire, the highest TCR clonality was observed in the case of CTCs under the cut-off and a positive ELISpot response (p=0.03). Whereas, at follow-up, patients showing a good clinical response coupled with a low number of CTCs were characterized by the most elevated TCR clonality (p<0.05). The detection of CTCs≥6 in at least one time-point was associated with a lower TCR clonality (p=0.02). Intriguingly, by combining overall survival analysis with TCR repertoire, we highlighted a potential prognostic role of the TCR clonality measured at follow-up (p=0.03). Conclusion: These data, whether validated in a larger cohort of patients, suggest that the combined analysis of CTCs and circulating anti-tumor T-cell immunity could represent a valuable immune-oncological biomarker for the liquid biopsy field. The clinical application of this promising tool could improve the management of mBC patients, especially in the setting of immunotherapy, a rising approach for BC treatment requiring reliable predictive biomarkers

Dysmetabolic circulating tumor cells are prognostic in metastatic breast cancer

Circulating tumor cells (CTCs) belong to a heterogeneous pool of rare cells, and a unequivocal phenotypic definition of CTC is lacking. Here, we present a definition of metabolically-altered CTC (MBA-CTCs) as CD45-negative cells with an increased extracellular acidification rate, detected with a single-cell droplet microfluidic technique. We tested the prognostic value of MBA-CTCs in 31 metastatic breast cancer patients before starting a new systemic therapy (T0) and 3\u20134 weeks after (T1), comparing results with a parallel FDA-approved CellSearch (CS) approach. An increased level of MBA-CTCs was associated with: I) a shorter median PFS pre-therapy (123 days vs. 306; p < 0.0001) and during therapy (139 vs. 266 days; p = 0.0009); ii) a worse OS pre-therapy (p = 0.0003, 82% survival vs. 20%) and during therapy (p = 0.0301, 67% survival vs. 38%); iii) good agreement with therapy response (kappa = 0.685). The trend of MBA-CTCs over time (combining data at T0 and T1) added information with respect to separate evaluation of T0 and T1. The combined results of the two assays (MBA and CS) increased stratification accuracy, while correlation between MBA and CS was not significant, suggesting that the two assays are detecting different CTC subsets. In conclusion, this study suggests that MBA allows detection of both EpCAM-negative and EpCAM-positive, viable and label-free CTCs, which provide clinical information apparently equivalent and complementary to CS. A further validation of proposed method and cut-offs is needed in a larger, separate study

Re-Imagining Resilient Productive Landscapes: Perspectives from Planning History

This book explores how lessons from past urban planning experiences can inform current debates on urban agriculture. Productive landscapes today have been posited as instruments for the positive transformation related to territorial fragility and abandonment, promoting social cohesion, food security and wider environmental and economic benefits. The book will re-map the way in which seeming landscape limitations and challenges can be turned into potential, innovation and a new lease of urban-rural life. It does so by drawing on significant past urban agricultural experiences in planning as vectors for new critical reflections relevant to re-igniting ideas for future envisioning of urban scenarios in which productive landscapes play fundamental transformative roles. The focus is on planning ideas and the roles of key individual planners, all of which have designed agricultural strategies for the city at some point in their careers. It intends to help us today reimagine urban-rural relationships, and the transformation of under or mis-used urban open spaces, peri-urban areas, fringe conditions and in-between spaces

Development of a simulation model of the rotary tube piercing process and fem application to improve the quality of seamless tubes

This research focuses on the understanding and FEM modelling of the phenomena that affect a rotary tube piercing process or Mannesmann process and has been developed in cooperation with a company (TENARIS-Dalmine S.p.A.). The work has been divided into two parts. In the first one the 2D FEM model definition and implementation is presented. In this phase the aspects related to the schematization of a three-dimensional process into a two-dimensional one were discussed with the Company. The second part deals with the simulative analysis of the hole beginning and propagation, and the comparison between simulation and experimental results. This comparison allowed the critical damage value to be evaluated, for the materials used by the Company, and identified the beginning of the bar failure (Mannesmann effect) in the simulations. The material breakage was studied using a customized version of the commercial finite element code DEFORM 2D. The good agreement between simulation results and experimental tests shows the capability of the FEM code to furnish good indications in the optimization of a rotary tube piercing process

Liquid biopsy: a review

The access to tumor material from a simple blood sample is generally called liquid biopsy. In this review, we describe the needs for such a technique, and the potential applications in oncology. We describe different classes of targets of liquid biopsy: circulating tumor cells, circulating tumor DNA, and exosomes, presenting current evidence, clinical meaning and utility of each class. Then, we explore in more depth methods for the characterization of CTC and ctDNA -the two targets actually considered closer to the bedside-, from a genetic, epigenetic and, in case of CTC, proteomic point of view. We compare applications, promises and limitations of CTC and ctDNA in the clinical setting. Finally, we offer a perspective on the clinical evaluation of genetic characterization from liquid biopsy samples, taking into consideration that certain mutations — often considered to be malignant — have been frequently described in healthy subjects

Long Noncoding RNAs as Innovative Urinary Diagnostic Biomarkers

The characterization of circulating tumor cells (CTCs) is now widely studied as a promising source of cancer-derived biomarkers because of their role in tumor formation and progression. However, CTCs analysis presents some limitations and no standardized method for CTCs isolation from urine has been defined so far. In fact, besides blood, urine represents an ideal source of noninvasive biomarkers, especially for the early detection of genitourinary tumors. Besides CTCs, long noncoding RNAs (lncRNAs) have also been proposed as potential noninvasive biomarkers, and the evaluation of the diagnostic accuracy of urinary lncRNAs has dramatically increased over the last years, with many studies being published. Therefore, this review provides an update on the clinical utility of urinary lncRNAs as novel biomarkers for the diagnosis of bladder and prostate cancers