Biological and clinical perspectives of the actionable gene fusions and amplifications involving tyrosine kinase receptors in lung cancer

MS is supported by a Juan-Rodés contract from the Instituto de Salud Carlos III (JR20/00015).This work was supported by Spanish grant from ICAPEM (ICAPEM becas 2021), to EP, and by Spanish grant form Asociación Española Contra el Cancer (AECC) (grant number GCB14142170MONT) to MSC. We thank M Rey providing language help and O Romero for data analysis support.Identifying molecular oncogenic drivers is crucial for precision oncology. Genetic rearrangements, including gene fusions and gene amplification, involving and activating receptor tyrosine kinases (RTKs) are recurrent in solid tumors, particularly in non-small cell lung cancer. Advances in the tools to detect these alterations have deepened our understanding of the underlying biology and tumor characteristics and have prompted the development of novel inhibitors targeting activated RTKs. Nowadays, druggable oncogenic rearrangements are found in around 15% of lung adenocarcinomas. However, taken separately, each of these alterations has a low prevalence, which poses a challenge to their diagnosis. The identification and characterization of novel targetable oncogenic rearrangements in lung cancer continue to expand, as shown by the recent discovery of the CLIP1-LTK fusion found in 0.4% of lung adenocarcinomas. While tyrosine kinase inhibitors that block the activity of RTKs have represented a breakthrough in the therapeutic landscape by improving the prognosis of this disease, prolonged treatment inevitably leads to the development of acquired resistance. Here, we review the oncogenic fusions and gene amplifications involving RTK in lung cancer. We address the genetic and molecular structure of oncogenic RTKs and the methods to diagnose them, emphasizing the role of next-generation sequencing technologies. Furthermore, we discuss the therapeutic implications of the different tyrosine kinase inhibitors, including the current clinical trials and the mechanisms responsible for acquired resistance. Finally, we provide an overview of the use of liquid biopsies to monitor the course of the disease

MYC activation impairs cell-intrinsic IFNγ signaling and confers resistance to anti-PD1/PD-L1 therapy in lung cancer

Elucidating the adaptive mechanisms that prevent host immune response in cancer will help predict efficacy of anti-programmed death-1 (PD1)/L1 therapies. Here, we study the cell-intrinsic response of lung cancer (LC) to interferon-y (IFNy), a cytokine that promotes immunoresponse and modulates programmed death-ligand 1 (PD-L1) levels. We report complete refractoriness to IFNy in a subset of LCs as a result of JAK2 or IFNGR1 inactivation. A submaximal response affects another subset that shows constitutive low levels of IFNy-stimulated genes (IySGs) coupled with decreased H3K27ac (histone 3 acetylation at lysine 27) depo-sition and promoter hypermethylation and reduced IFN regulatory factor 1 (IRF1) recruitment to the DNA on IFNy stimulation. Most of these are neuroendocrine small cell LCs (SCLCs) with oncogenic MYC/MYCL1/ MYCN. The oncogenic activation of MYC in SCLC cells downregulates JAK2 and impairs IySGs stimulation by IFNy. MYC amplification tends to associate with a worse response to anti-PD1/L1 therapies. Hence alterations affecting the JAK/STAT pathway and MYC activation prevent stimulation by IFNy and may predict anti-PD1/L1 efficacy in LC

SMARCA4 deficient tumours are vulnerable to KDM6A/UTX and KDM6B/JMJD3 blockade

The authors thank Isabel Bartolessis (Cancer Genetics Group) at IJC for technical assistance. This work was supported by the Spanish Ministry of Economy and CompetitivityMINECO (grant number SAF-2017-82186R, to M.S.-C., and grant PI19/01320 to A. Villanueva) and from the Fundacion Cientifica of the Asociacion Espanola Contra el Cancer (AECC) (grant number GCB14142170MONT) to M.S.-C. A. Villanueva is also funded by the Department of Health of the Generalitat de Catalunya (2014SGR364). O.A. R. received a Juan de la Cierva postdoctoral contract (grant No. IJCI-2016-28201, until November 2019) and an AECC research contract (INVES19045ROME from December 2019). A. Vilarrubi, P.L. and A.A. are supported by pre-doctoral contracts from the Spanish MINECO (FPI-fellowship: PRE2018-084624, BES-2015-072204 and FPU17/00067). M.S. was supported by a Rio Hortega contract from the Instituto de Salud Carlos III (CM17/00180). L.F. received a European Union Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie Actions grant agreement, number 799850.Despite the genetic inactivation of SMARCA4, a core component of the SWI/SNF-complex commonly found in cancer, there are no therapies that effectively target SMARCA4-deficient tumours. Here, we show that, unlike the cells with activated MYC oncogene, cells with SMARCA4 inactivation are refractory to the histone deacetylase inhibitor, SAHA, leading to the aberrant accumulation of H3K27me3. SMARCA4-mutant cells also show an impaired transactivation and significantly reduced levels of the histone demethylases KDM6A/UTX and KDM6B/JMJD3, and a strong dependency on these histone demethylases, so that its inhibition compromises cell viability. Administering the KDM6 inhibitor GSK-J4 to mice orthotopically implanted with SMARCA4-mutant lung cancer cells or primary small cell carcinoma of the ovary, hypercalcaemic type (SCCOHT), had strong anti-tumour effects. In this work we highlight the vulnerability of KDM6 inhibitors as a characteristic that could be exploited for treating SMARCA4-mutant cancer patients.Spanish Ministry of Economy and Competitivity-MINECO SAF-2017-82186R PI19/01320Fundacion Cientifica of the Asociacion Espanola Contra el Cancer (AECC) GCB14142170MONTDepartment of Health of the Generalitat de Catalunya 2014SGR364Juan de la Cierva postdoctoral contract IJCI-2016-28201AECC research contract INVES19045ROMESpanish MINECO PRE2018-084624 BES-2015-072204 FPU17/00067Instituto de Salud Carlos III European Commission CM17/00180European Union Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie Actions grant agreement 79985

Efficacy of CDK4/6 inhibitors in preclinical models of malignant pleural mesothelioma

Background There is no effective therapy for patients with malignant pleural mesothelioma (MPM) who progressed to platinum-based chemotherapy and immunotherapy. Methods We aimed to investigate the antitumor activity of CDK4/6 inhibitors using in vitro and in vivo preclinical models of MPM. Results Based on publicly available transcriptomic data of MPM, patients with CDK4 or CDK6 overexpression had shorter overall survival. Treatment with abemaciclib or palbociclib at 100 nM significantly decreased cell proliferation in all cell models evaluated. Both CDK4/6 inhibitors significantly induced G1 cell cycle arrest, thereby increasing cell senescence and increased the expression of interferon signalling pathway and tumour antigen presentation process in culture models of MPM. In vivo preclinical studies showed that palbociclib significantly reduced tumour growth and prolonged overall survival using distinct xenograft models of MPM implanted in athymic mice. Conclusions Treatment of MPM with CDK4/6 inhibitors decreased cell proliferation, mainly by promoting cell cycle arrest at G1 and by induction of cell senescence. Our preclinical studies provide evidence for evaluating CDK4/6 inhibitors in the clinic for the treatment of MPM

PERBEDAAN KEKUATAN RETENSI ANTARA KAITAN MAGNET DAN KAITAN STUD (MDI) PADA OVERDENTURE RAHANG BAWAH (Penelitian Eksperimental Laboratoris)

Overdenture is a partial denture or full denture that covers all the root surface or implant. Overdenture can use tooth, tooth root or implant as an abutment. An attachment can be used to add more retention. Magnet and stud attachment is often used for this purpose. The aim of this research is to know the retentional force difference between overdenture using magnet attachment and stud attachment (MDI). The study was conducted on 10 acrylic plates size 50x10x10 cm for each attachment. Magnet (MAGFIT TM DX) dan stud attachment (MDI IMTEC 3M) were used on lower jaw edentulous model made from acrylic. Magnet was then attached to acrylic plate and the keeper attached to edentulous model, on stud atachment (MDI) the male attachment (implant) was attached to edentulous model and the female attachment (metal housing) attached to acrylic plate. The overdenture was then attached to the modified Van Kampen. Retention was then measured by pulling the acrylic plate which has been attached to edentulous model. Statistical method using independent t-test with significance α = 0.05. The result showed that there was a significant difference between overdenture using stud attachment (MDI) and overdenture using magnet attachmet (p<0.05). In conclusion, the overdenture using stud attachment provide more retention than overdenture using magnet attachment

Satisfacción, salud y estrés laboral del profesorado universitario según su situación contractual

Objetivo Analizar los efectos de la estabilidad contractual de los profesores universitarios en las dimensiones psicosociales de satisfacción, salud y estrés.Método Estudio descriptivo cuasi-experimental, tomando como muestra a 145 profesores de siete universidades públicas catalanas. El estudio se realizó durante el mes de abril de 2014. Se utilizó como base el cuestionario ISTAS21, versión media, en lengua castellana, del Cuestionario Psicosocial de Copenhaguen. De las 21 escalas incluidas, sólo se han tenido en cuenta las siete escalas relacionadas con los factores evaluados. Se realizaron una serie de análisis de varianza (ANOVA) considerando las puntuaciones obtenidas en las tres dimensiones psicosociales evaluadas como variables dependientes y la estabilidad laboral (permanencia y dedicación laboral) como variable independiente.Resultados Los profesores de tiempo completo con contratos laborales temporales muestran una peor salud física y mental —a pesar de ser el grupo más joven—, mayores síntomas de estrés y una menor satisfacción laboral, en comparación con profesores a tiempo completo y contrato estable; o profesores de tiempo parcial.Conclusiones La estabilidad laboral tiene repercusiones en la salud y calidad de vida laboral de los profesores que se encuentran en situación de inestabilidad, teniendo ello además repercusiones en la docencia y la investigación.Objective There’s a lot of literature about teachers’ work conditions and the effects of such conditions on them. The goal of this research focuses on the analysis of the effects derived from the type of labour relationship with the University on the psycho-social dimensions of  satisfaction, stress and perceived health.Methods Quasi-experimental study on a sample of 145 university professors from the seven public universities from Catalonia (Spain). The data was obtained in April 2014, by means of ISTAS21 questionnaire, intermediate version, in Spanish. From the 21 subscales in the instrument, only seven were used for data-gathering since they encompassed the dimensions of interest. A series of ANOVA analyses were performed, taking the three psycho-social dimensions as dependent variables and work stability and dedication (e.g. full- vs. partial-time) as independent variables.Results Professors with full-time work and unstable contracts showed worse health (both physical and mental) despite being the youngest, as well as larger symptoms of stress and lower job-satisfaction, compared with full-time professors with stable contracts and even compared part-time-jobs professors.Conclusion Job-stability seems to be a central variable in work-quality and health for the professors whose labour situation is unstable, something that influences both their teaching and research quality

Modulation of aberrant NF1 pre-mRNA splicing by kinetin treatment

Neurofibromatosis type 1 is one of the most common neurocutaneous autosomal dominant disorders. It is caused by mutations in the neurofibromatosis type 1 (NF1) gene and approximately 30–40% of them affect the correct splicing of NF1 pre-mRNA. In this report, we evaluate the effect of five different drugs, previously found to modify splicing in several genetic disorders, on the splicing of mutated NF1 alleles. For this purpose, cell lines derived from patients bearing 19 different NF1-splicing defects were used. Our results showed that kinetin partially corrects the splicing defect in four of the studied mutations (c.910C>T, c.3113G>A, c.6724C>T and c.6791dupA). Our study is a valuable contribution to the field because it identifies new exon-skipping events that can be reversed by kinetin treatment and provides new information about kinetin splicing modulation. However, owing to the nature of mutations in our patients, kinetin treatment could not be used as a therapeutic agent in these cases

Deep analysis of acquired resistance to FGFR1 inhibitor identifies MET and AKT activation and an expansion of AKT1 mutant cells

The development of acquired resistance (AR) to tyrosine kinase inhibitors (TKIs) of FGFR1 activation is currently not well understood. To gain a deeper insight into this matter in lung cancer, we used the FGFR1-amplified DMS114 cell line and generated multiple clones with AR to an FGFR1-TKI. We molecularly scrutinized the resistant cells, using whole-exome sequencing, RNA sequencing and global DNA methylation analysis. Our results show a de novo activation of AKT and ERK, and a reactivation of mTOR. Furthermore, the resistant cells exhibited strong upregulation and activation of MET, indicating crosstalk between the FGFR1 and MET axes. The resistant cells also underwent a global decrease in promoter hypermethylation of the CpG islands. Finally, we observed clonal expansion of a pre-existing change in AKT1, leading to S266L substitution, within the kinase domain of AKT. Our results demonstrate that AR to FGFR1-TKI involves deep molecular changes that promote the activation of MET and AKT, coupled with common gene expression and DNA methylation profiles. The expansion of a substitution at AKT1 was the only shared genetic change, and this may have contributed to the AR.This work was supported by Spanish grants SAF2014-54571-R (to M Sanchez-Cespedes), AGAUR (Agency for Management of University and Research Grants) (2014SGR641) (to M Sanchez-Cespedes), and PTA2014-09515 (to M Dabad). The CNAG-CRG laboratory is a member of the Spanish National Bioinformatics Institute (INB), PRB2-ISCIII and is supported by grants PT13/0001, from the PE I+D+i 2013-2016, funded by ISCIII and FEDER (to A Esteve-Codina)