Assessment of the Royal Decree 244/2019 in the Spanish Electrical Regulatory Framework considering power quality issues related to harmonic distortion associated to nonlinear loads in grid-connected microgrids

Postprint (published version

Tumor innate immunity primed by specific interferon-stimulated endogenous retroviruses.

Mesenchymal tumor subpopulations secrete pro-tumorigenic cytokines and promote treatment resistance1-4. This phenomenon has been implicated in chemorefractory small cell lung cancer and resistance to targeted therapies5-8, but remains incompletely defined. Here, we identify a subclass of endogenous retroviruses (ERVs) that engages innate immune signaling in these cells. Stimulated 3 prime antisense retroviral coding sequences (SPARCS) are oriented inversely in 3' untranslated regions of specific genes enriched for regulation by STAT1 and EZH2. Derepression of these loci results in double-stranded RNA generation following IFN-γ exposure due to bi-directional transcription from the STAT1-activated gene promoter and the 5' long terminal repeat of the antisense ERV. Engagement of MAVS and STING activates downstream TBK1, IRF3, and STAT1 signaling, sustaining a positive feedback loop. SPARCS induction in human tumors is tightly associated with major histocompatibility complex class 1 expression, mesenchymal markers, and downregulation of chromatin modifying enzymes, including EZH2. Analysis of cell lines with high inducible SPARCS expression reveals strong association with an AXL/MET-positive mesenchymal cell state. While SPARCS-high tumors are immune infiltrated, they also exhibit multiple features of an immune-suppressed microenviroment. Together, these data unveil a subclass of ERVs whose derepression triggers pathologic innate immune signaling in cancer, with important implications for cancer immunotherapy

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

MET and epithelial to mesenchymal transition as novel targets in small cell lung cancer

Small cell lung carcinoma (SCLC) is a highly lethal disease due to its chemorefractory nature after first line treatment. The mechanisms to overcome this resistance have remained elusive to tackle up to date. MET is a transmembrane tyrosine kinase receptor and its activation is associated with increased motility, migration and invasion in cancer cells. MET is activated in several tumour types and has been linked to patient prognosis. MET activation by its natural ligand Hepatocyte Growth Factor (HGF) has been involved in Epithelial to Mesenchymal Transition (EMT), a process by which cells decrease adhesion, lose polarity, acquire the ability to migrate and invade surrounding tissue. EMT is also associated with resistance to anticancer agents. We hypothesized that HGF-induced EMT would explain resistance to chemotherapy in SCLC and that MET inhibitors could revert this phenomenon. In our work we demonstrated that HGF-induced MET activation in SCLC cells resulted in a more aggressive phenotype. In human SCLC we demonstrated in two independent series that MET phosphorylation was associated with poor prognosis. In preclinical models we observed that MET activation by HGF induced EMT that resulted in chemoresistance in vitro and in vivo. MET inhibition was able to block or reverse this process and resensitized cells to chemotherapy. Mesenchymal markers in human SCLC specimens were associated with MET activation, predicted a worse survival and were upregulated in chemorefractory disease. Finally, increased HGF serum levels in SCLC patients correlated with higher risk of death. These results suggest that the use of MET inhibitors in combination with chemotherapy as a therapeutic approach in the MET-activated subpopulation of SCLC merits further investigation.El Cáncer de Pulmón de Célula Pequeña (CPCP) es una enfermedad altamente letal debido a su naturaleza quimiorefractaria después del tratamiento de primera línea. Los mecanismos para derrotar estar quimioresistencia han fracasado hasta la fecha. MET es un receptor de membrana tirosina cinasa y su activación está asociada a una incrementada motilidad, migración e invasión en células tumorales. MET está activado en diversos tipos tumorales y está asociado al pronóstico de los pacientes. La activación de MET mediante su ligando natural “Factor de Crecimiento Hepatocitario” (HGF) ha estado involucrado en la Transición Epitelio- Mesenquimal (EMT), un proceso mediante el cual las células disminuyen su adhesión, pierden la polaridad, adquieren la habilidad de migrar e invaden los tejidos adyacentes. La EMT está también asociada a la resistencia a agentes antitumorales. Nuestra hipótesis es que la EMT inducida por HGF explicaría la resistencia a la quimioterapia en el CPCP y que los inhibidores de MET podrían revertir este fenómeno. En nuestro trabajo demostramos que la activación de MET inducida por HGF en líneas celulares de CPCP dio lugar a un fenotipo más agresivo. En dos series independientes de CPCP humano demostramos que la fosforilación de MET estaba asociada a un peor pronóstico. En modelos preclínicos observamos que la activación de MET mediante HGF indujo la EMT, dando lugar a quimioresistencia in vitro e in vivo. La inhibición de MET fue capaz de bloquear o revertir este proceso, sensibilizando las células tumorales a la quimioterapia. Los marcadores mesenquimales en muestras humanas de CPCP se asociaron a activación de MET, prediciendo una peor supervivencia. Además, la expresión de estos marcadores estaba incrementada en la enfermedad quimiorefractaria. Estos resultados sugieren que el uso de inhibidores de MET en combinación con quimioterapia como una aproximación terapéutica en la subpoblación de pacientes de CPCP con activación de MET merece una investigación más extensa

High circulating hepatocyte growth factor levels associate with epithelial to mesenchymal transition and poor outcome in small cell lung cancer patients

We have previously shown that Met activation through the hepatocyte growth factor (HGF) increases tumorogenesis, induces epithelial-to-mesenchymal transition (EMT) and chemoresistance in SCLC. We sought to evaluate circulating HGF levels in SCLC patients and assess correlation with outcome and EMT features in the tumor. Serum samples from patients with SCLC were prospectively obtained at diagnosis, response evaluation and progression. HGF serum (sHGF) was quantified by ELISA. EMT markers and p-Met/Met were assayed by immunohistochemistry in tumor samples. Clinical data were prospectively recorder. One-hundred twelve patients were included. High baseline levels of sHGF were associated with shorter overall survival (p=0.007) and remained independently associated with survival in the multivariate analysis (p=0.016). For stage IV patients, an increase of sHGF levels at response evaluation (p=0.042) and at progression (p=0.003) were associated with poor outcome. sHGF levels were associated (p<0.05) with a mesenchymal phenotype in the tumor. In conclusion, high sHGF at diagnosis and increases during the course of the disease predict for poor outcome in SCLC patients and associate with EMT in the tumor. These data provide novel evidence on a role of sHGF in the adverse clinical behavior of SCLC and support testing Met inhibitors in patients with high sHGF

MET expression and copy number heterogeneity in nonsquamous non-small cell lung cancer (nsNSCLC).

OBJECTIVE: We aimed to assess MET intratumoral heterogeneity and its potential impact on biomarker-based patient selection as well as potential surrogate biomarkers of MET activation. METHODS: Our study included 120 patients with non-squamous Non-small-cell Lung Cancer (nsNSCLC), of which 47 were incorporated in tissue microarrays (TMA). Four morphologically distinct tumor areas were selected to assess MET heterogeneity. MET positivity by immunohistochemistry (IHC) was defined as an above-median H-score and by +2/+3 staining intensity in >50% of tumor cells (Metmab criteria). MET FISH positivity was defined by MET/CEP7 ratio ≥ 2.0 and/or MET ≥ 5.0. MET staining pattern (cytoplasmic vs. membranous) and mesenchymal markers were investigated as surrogates of MET activation. RESULTS: Median MET H-score was 140 (range 0-400) and 47.8% of patients were MET positive by Metmab criteria. Eight cases (6.8%) were MET FISH positive and showed higher H-scores (p = 0.021). MET positivity by IHC changed in up to 40% of cases among different tumor areas, and MET amplification in 25-50%. Cytoplasmic MET staining and positivity for vimentin predicted poor survival (p = 0.042 and 0.047, respectively). CONCLUSIONS: MET status is highly heterogeneous among different nsNSCLC tumor areas, hindering adequate patient selection for MET-targeted therapies. MET cytoplasmic staining and vimentin might represent surrogate markers for MET activation.This work was supported by RD12/0036/0051 /FEDER, PI13/00140/FEDER, a grant from Fundacio Marato de TV3. Ref.666/C/2013 and 2014 SGR 740. the “Xarxa de Bancs de tumors sponsored by Pla Director d’Oncologia de Catalunya (XBTC).” JA is recipient of intensification programme ISCIII

The First-in-class Anti-EGFR Antibody Mixture Sym004 Overcomes Cetuximab Resistance Mediated by EGFR Extracellular Domain Mutations in Colorectal Cancer.

PURPOSE: Approved anti-EGFR antibodies cetuximab and panitumumab provide significant clinical benefit in patients with metastatic colorectal cancer (MCRC). However, patients ultimately develop disease progression, often driven by acquisition of mutations in the extracellular domain (ECD) of EGFR. Sym004 is a novel 1:1 mixture of two nonoverlapping anti-EGFR mAbs that recently showed promising clinical activity in a phase I trial in MCRC. Our aim was to determine the efficacy of Sym004 to circumvent cetuximab resistance driven by EGFR ECD mutations. EXPERIMENTAL DESIGN: Functional studies were performed to assess drug-receptor binding as well as ligand-dependent activation of individual EGFR mutants in the presence of cetuximab, panitumumab, and Sym004. Cell viability and molecular effects of the drugs were assayed in cetuximab-resistant cell lines and in tumor xenograft models. Efficacy of Sym004 was evaluated in patients progressing to cetuximab that harbored EGFR mutation in the post-cetuximab tumor sample. RESULTS: Contrary to cetuximab and panitumumab, Sym004 effectively bound and abrogated ligand-induced phosphorylation of all individual EGFR mutants. Cells resistant to cetuximab harboring mutations in EGFR maintained sensitivity to Sym004, which was consistent with an effective suppression of EGFR downstream signaling, translating into profound and sustained tumor regression in the xenograft model. As proof-of-principle, a patient with a tumor harboring an EGFR mutation (G465R) following cetuximab therapy benefited from Sym004 therapy. CONCLUSIONS: Sym004 is an active drug in MCRC resistant to cetuximab/panitumumab mediated by EGFR mutations. EGFR mutations are potential biomarkers of response to Sym004 to be evaluated in ongoing large clinical trials.This work was supported by PI12/00989, PI15/00457, DTS15/00048, RD12/0036/0051, 2014SGR740, PIE15/00008, and by the Xarxa de Banc de Tumors de Cataluny