ALK-fusion transcripts can be detected in extracellular vesicles (EVs) from nonsmall cell lung cancer cell lines and patient plasma: Toward EV-based noninvasive testing

Artículo escrito por un elevado número de autores, sólo se referencian el que aparece en primer lugar y los autores pertenecientes a la UAMALK rearrangements are present in 5% of nonsmall cell lung cancer (NSCLC) tumors and identify patients who can benefit from ALK inhibitors. ALK fusions testing using liquid biopsies, although challenging, can expand the therapeutic options for ALK-positive NSCLC patients considerably. RNA inside extracellular vesicles (EVs) is protected from RNases and other environmental factors, constituting a promising source for noninvasive fusion transcript detection. EVs from H3122 and H2228 cell lines, harboring EML4-ALK variant 1 (E13; A20) and variant 3 (E6a/b; A20), respectively, were successfully isolated by sequential centrifugation of cell culture supernatants. EVs were also isolated from plasma samples of 16 ALK-positive NSCLC patients collected before treatment initiation. Purified EVs from cell cultures were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and flow cytometry. Western blot and confocal microscopy confirmed the expression of EV-specific markers as well as the expression of EML4-ALK-fusion proteins in EV fractions from H3122 and H2228 cell lines. In addition, RNA from EV fractions derived from cell culture was analyzed by digital PCR (dPCR) and ALK-fusion transcripts were clearly detected. Similarly, plasma-derived EVs were characterized by NTA, flow cytometry, and the ExoView platform, the last showing that EV-specific markers captured EV populations containing ALK-fusion protein. Finally, ALK fusions were identified in 50% (8/16) of plasma EV-enriched fractions by dPCR, confirming the presence of fusion transcripts in EV fractions. ALK-fusion transcripts can be detected in EV-enriched fractions. These results set the stage for the development of EV-based noninvasive ALK testingThis study has been funded by Instituto de Salud Carlos III through the project “PI17/01977” (co-funded by European Regional Development Fund/European Social Fund “A way to make Europe”/“Investing in your future”). The work presented in this paper also received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No 875160. E. Sa´nchezHerrero was funded by the Consejerıa de Ciencia, Universidades e Innovacio´n of the Comunidad de Madrid (Doctorados Industriales of the Comunidad de Madrid IND2019/BMD-17258). R. Serna-Blasco was funded by the Consejerıa de Educacio´n, Juventud y Deporte of the Comunidad de Madrid and by the Fondo Social Europeo (Programa Operativo de Empleo Juvenil, and Iniciativa de Empleo Juvenil, PEJD2018-PRE/BMD-8640). M. Vales-Go´mez was funded by Spanish Ministry of Science and Innovation (MCIU/AEI/FEDER, EU) grant RTI2018-093569-B-I00 and Madrid Regional Government under grant “IMMUNOTHERCAN” (S2017/BMD-3733-2). This work was supported by GEIVEX, Network of Excellence in the Research and Innovation on Exosomes REDIEX, SAF2015-71231-REDT, Network of Excellence in Translational NeTwork for the CLinical application of Extracellular VesicleS (TeNTaCLES) (RED2018-102411-T). M. Provencio, grants from BristolMyers Squibb, Roche, and AstraZenec

Lenalidomide plus R-GDP (R2-GDP) in relapsed/ refractory diffuse large B-cell lymphoma: final results of the R2-GDP-GOTEL trial and immune biomarker subanalysis

Artículo escrito por un elevado número de autores, solo se referencian el que aparece en primer lugar, el nombre del grupo de colaboración, si le hubiere, y los autores pertenecientes a la UAMPurpose: New therapeutic options are needed in relapsed/refracResults: After a median follow-up of 37 months, ORR was tory diffuse large B-cell lymphoma (R/R DLBCL). Lenalidomide-60.2% [37.1% complete responses (CR) and 23.1% partial based schedules can reverse rituximab refractoriness in lymphoma. responses (PR)]. Median OS was 12 months (47 vs. 6 months Patients and Methods: In the phase II R2-GDP trial, 78 in CR vs. no CR); median PFS was 9 months (34 vs. 5 months in patients unsuitable for autologous stem cell transplant received CR vs. no CR). In the primary refractory population, ORR was treatment with the following schedule: lenalidomide 10 mg Days 45.5% (21.2% CR and 24.3% PR). Most common grade 3–4 (D)1–14, rituximab 375 mg/m2 D1, cisplatin 60 mg/m2 D1, adverse events were thrombocytopenia (60.2%), neutropenia gemcitabine 750 mg/m2 D1 and D8, and dexamethasone 20 mg (60.2%), anemia (26.9%), infections (15.3%), and febrile neutroD1–3, up to 6 cycles (induction phase), followed by lenalidomide penia (14.1%). Complete responses were associated with a sharp 10 mg (or last lenalidomide dose received) D1–21 every 28 days decrease in circulating myeloid-derived suppressor cells and (maintenance phase). Primary endpoint was overall response regulatory T cells. rate (ORR). Secondary endpoints included progression-free Conclusions: R2-GDP schedule is feasible and highly active survival (PFS), overall survival (OS), safety, and monitorization in R/R DLBCL, including the primary refractory population. of key circulating immune biomarkers (EU Clinical Trials Reg-Immune biomarkers showed differences in responders versus ister number: EudraCT 2014-001620-29). progressorsThis research was funded by the Spanish Lymphoma Oncology Group (GOTEL) with the financial support of Celgene (Investigator Initiated Trials Program); no grant numbers applicable. L. Hontecillas-Prieto is supported by the Consejería de Salud y Familias, Junta de Andalucía (RH-0047-2021). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fac

Determination of essential biomarkers in lung cancer: a real-world data study in Spain with demographic, clinical, epidemiological and pathological characteristics

Artículo escrito por un elevado número de autores, sólo se referencia el que aparece en primer lugar y los autores pertenecientes a la UAMThe survival of patients with lung cancer has substantially increased in the last decade by about 15%. This increase is, basically, due to targeted therapies available for advanced stages and the emergence of immunotherapy itself. This work aims to study the situation of biomarker testing in Spain. The Thoracic Tumours Registry (TTR) is an observational, prospective, registry-based study that included patients diagnosed with lung cancer and other thoracic tumours, from September 2016 to 2020. This TTR study was sponsored by the Spanish Lung Cancer Group (GECP) Foundation, an independent, scientific, multidisciplinary oncology society that coordinates more than 550 experts and 182 hospitals across the Spanish territory. Nine thousand two hundred thirty-nine patients diagnosed with stage IV non-small cell lung cancer (NSCLC) between 2106 and 2020 were analysed. 7,467 (80.8%) were non-squamous and 1,772 (19.2%) were squamous. Tumour marker testing was performed in 85.0% of patients with non-squamous tumours vs 56.3% in those with squamous tumours (p-value < 0.001). The global testing of EGFR, ALK, and ROS1 was 78.9, 64.7, 35.6% respectively, in non-squamous histology. PDL1 was determined globally in the same period (46.9%), although if we focus on the last 3 years it exceeds 85%. There has been a significant increase in the last few years of all determinations and there are even close to 10% of molecular determinations that do not yet have targeted drug approval but will have it in the near future. 4,115 cases had a positive result (44.5%) for either EGFR, ALK, KRAS, BRAF, ROS1, or high PDL1. Despite the lack of a national project and standard protocol in Spain that regulates the determination of biomarkers, the situation is similar to other European countries. Given the growing number of different determinations and their high positivity, national strategies are urgently needed to implement next-generation sequencing (NGS) in an integrated and cost-effective way in lung cancerThis study was supported by the European Union Horizon 2020 Research and Innovation Program under grant agreement n° 875160 CLARIFY projec

Genomic analyses of microdissected Hodgkin and Reed-Sternberg cells: mutations in epigenetic regulators and p53 are frequent in refractory classic Hodgkin lymphoma

This work was supported by grants from the Plan Nacional de I + D + I cofinanced by the ISCIII-Subdirección General de Evaluación and the Fondo Europeo de Desarrollo Regional (FEDER), PI12/1832, the Spanish Association for Cancer Research (AECC), and Programas para Grupos de Investigación de la Comunidad Autónoma de Madrid (Biomedicina 2017)

Machine learning estimated probability of relapse in early-stage non-small-cell lung cancer patients with aneuploidy imputation scores and knowledge graph embeddings

Low-stage lung cancer is known to recur unpredictably, and patients receiving various treatment methods like radiation, chemotherapy, and immunotherapies have been seen to respond very differently. Identifying a priori if a patient is going to relapse or not could make a difference in terms of saving lives and personalized care offered. In this work, we provide an answer to the following research question: Is it possible to enhance the machine learning (ML) of the estimated probability of relapse in early-stage non-small-cell lung cancer (NSCLC) patients with aneuploidy imputation scores? To predict recurrence in 1,348 early-stage (I–II) NSCLC patients, we train graph ML models utilizing the Spanish pulmonary cancer group knowledge graph enriched with triples from pathway imputation. ML models trained on Knowledge graph data enriched with triples from pathway score imputation present an 82% Precision and 91% Specificity in predicting relapse over 200 patients from a held-out test set. ML models trained using graphs data could prove useful supplemental tool in the TNM classification systems and improve a lung cancer patient's prognosisThis paper is part of the CLARIFY project funded by the EU’s Horizon 2020 Research and Innovation Program (grant No. 875160

Circulating tumor DNA as a cancer biomarker: An overview of biological features and factors that may impact on ctDNA analysis

Cancer cells release nucleic acids, freely or associated with other structures such as vesicles into body fluids, including blood. Among these nucleic acids, circulating tumor DNA (ctDNA) has emerged as a minimally invasive biomarker for tumor molecular profiling. However, certain biological characteristics of ctDNA are still unknown. Here, we provide an overview of the current knowledge about ctDNA biological features, including size and structure as well as the mechanisms of ctDNA shedding and clearance, and the physio-pathological factors that determine ctDNA levels. A better understanding of ctDNA biology is essential for the development of new methods that enable the analysis of ctDNAES-H was funded by the Consejerıa de Ciencia, Universidades e Innovación of the Comunidad de Madrid (Doctorados Industriales of the Comunidad de Madrid IND2019/BMD17258). RS-B was funded by the Ministerio de Ciencia e Innovación (Programa Estatal de Investigación, Desarrollo e Innovacion Orientada a los Retos de la Sociedad, RTC2019-007359-1

CtDNA from body fluids is an adequate source for EGFR biomarker testing in advanced lung adenocarcinoma

Objectives: Epidermal growth factor receptor (EGFR) biomarker testing using blood-based liquid biopsies remains challenging due to the low concentration of circulating tumor DNA (ctDNA) in certain plasma samples. The aim of this study is to evaluate the usefulness for EGFR biomarker testing of ctDNA from pleural effusions, cerebrospinal fluids, ascites and pericardial effusions obtained during the clinical management of lung adenocarcinoma patients. Methods: For comparison purposes, 23 paired plasma and body fluid samples were collected from 17 patients with EGFR-positive lung adenocarcinoma. After circulating free DNA (cfDNA) isolation, samples were evaluated for the initial EGFR-sensitizing mutation and the p.T790M resistance mutation by array-based digital PCR (dPCR). Results: Body fluids had more cfDNA than plasma samples (1.90 vs. 0.36 ng/µL; p=0.0130), and more samples tested positive for EGFR mutations (21 vs. 16 samples), with a total of 28 vs. 22 variants detected. Furthermore, mutant allele frequencies (MAFs) observed in body fluids were significantly higher than those assessed in the paired plasma samples for EGFR-sensitizing mutations (median MAFs = 15.8 vs. 0.8%; p=0.0004) as well as for the p.T790M resistance mutation (median MAFs = 8.69 vs. 0.16%; p=0.0390). Importantly, two patients who had progressed on first-generation EGFR-tyrosine kinase inhibitors with a dubious result for p.T790M plasma (MAFs = 0.11%) had an indisputably positive result in their respective body fluid samples (MAFs = 10.25 and 9.66%). Conclusions: ctDNA derived from body fluids is an informative source for EGFR biomarker testing, with greater sensitivity than plasma samples.MB is supported by an i-PFIS predoctoral fellowship (Grant Number IFI18/00051) from ISCIII. ES is funded by the Consejería de Ciencia, Universidades e Innovación de la Comunidad de Madrid (Doctorados Industriales de la Comunidad de Madrid IND2019/BMD-17258), Spain

First-Line Biological Agents Plus Chemotherapy in Older Patients with Metastatic Colorectal Cancer: A Retrospective Pooled Analysis

Artículo escrito por un elevado número de autores, sólo se referencian el que aparece en primer lugar, el nombre del grupo de colaboración, si le hubiere, y los autores pertenecientes a la UAMBackground Biologicals, in combination with chemotherapy, are recommended as first-line treatment of metastatic colorectal cancer (mCRC); however, evidence guiding the appropriate management of older patients with mCRC is limited. Objective This study was undertaken to compare the efficacy and safety outcomes in older versus younger patients with mCRC who received first-line biological therapy. Methods This retrospective analysis used pooled data from five trials undertaken by the Spanish Cooperative Group for the Treatment of Digestive Tumours. All were studies of adults with advanced CRC who received first-line treatment with chemotherapy plus bevacizumab, cetuximab or panitumumab, stratified by age (≥ 65 vs. < 65 years). Endpoints included progression-free survival (PFS), overall survival (OS), overall response rate (ORR) and safety. Results In total, 999 patients from five studies were included in the analysis: 480 (48%) were aged ≥ 65 years, and 519 (52%) were aged < 65 years. Median PFS did not differ significantly between patients aged ≥ 65 and < 65 years (9.9 vs. 9.4 months; hazard ratio [HR] 1.01; 95% confidence interval [CI] 0.88–1.17). Median OS was significantly shorter in older than in younger patients (21.3 vs. 25.0 months; HR 1.21; 95% CI 1.04–1.41). There was no significant difference between older and younger patients in ORR (59 vs. 62%). Patients aged ≥ 65 years experienced significantly more treatment-related grade 3 or higher adverse events (61.67%) than did patients aged < 65 years (45.86%). Conclusions Biologicals plus chemotherapy is an effective first-line treatment option for selected patients aged ≥ 65 years with mCRC and has a manageable safety profile and efficacy comparable to that observed in younger patientsThis study was funded by the Spanish Cooperative Group for the Treatment of Digestive Tumours (TTD) collaborative group. All medical writing assistance was funded by the Spanish Cooperative TTD and Roche Farma S

Old and New Insights in the Treatment of Thyroid Carcinoma

Thyroid cancer is the endocrine tumor that bears the highest incidence with 33 550 new cases per year. It bears an excellent prognosis with a mortality of 1530 patients per year (Jemal et al.; 2007). We have been treating patients with thyroid carcinoma during many years without many innovations. Recently, we have assisted to the development of new agents for the treatment of this disease with unexpected good results. Here we present a review with the old and new methods for the treatment of this disease