Cancer is a rapidly evolving, multifactorial disease that accumulates numerous genetic and epigenetic alterations. This results in molecular and phenotypic heterogeneity within the tumor, the ...complexity of which is further amplified through specific interactions between cancer cells. We aimed to dissect the molecular mechanisms underlying the cooperation between different clones.
We produced clonal cell lines derived from the MDA-MB-231 breast cancer cell line, using the UbC-StarTrack system, which allowed tracking of multiple clones by color: GFP C3, mKO E10 and Sapphire D7. Characterization of these clones was performed by growth rate, cell metabolic activity, wound healing, invasion assays and genetic and epigenetic arrays. Tumorigenicity was tested by orthotopic and intravenous injections. Clonal cooperation was evaluated by medium complementation, co-culture and co-injection assays.
Characterization of these clones in vitro revealed clear genetic and epigenetic differences that affected growth rate, cell metabolic activity, morphology and cytokine expression among cell lines. In vivo, all clonal cell lines were able to form tumors; however, injection of an equal mix of the different clones led to tumors with very few mKO E10 cells. Additionally, the mKO E10 clonal cell line showed a significant inability to form lung metastases. These results confirm that even in stable cell lines heterogeneity is present. In vitro, the complementation of growth medium with medium or exosomes from parental or clonal cell lines increased the growth rate of the other clones. Complementation assays, co-growth and co-injection of mKO E10 and GFP C3 clonal cell lines increased the efficiency of invasion and migration.
These findings support a model where interplay between clones confers aggressiveness, and which may allow identification of the factors involved in cellular communication that could play a role in clonal cooperation and thus represent new targets for preventing tumor progression.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
CTCs have extensively been used for the monitoring and characterization of metastatic prostate cancer, but their application in the clinic is still very scarce. Besides, the resistance mechanisms ...linked to prostate cancer treatment remain unclear. Liquid biopsies represent the most promising alternative due to the complexity of biopsying bone metastasis and the duration of the disease. We performed a prospective longitudinal study in CTCs from 20 castration-resistant prostate cancer patients treated with docetaxel. For that, we used CellSearch® technology and a custom gene expression panel with qRT-PCR using a CTCs negative enrichment approach. We found that CTCs showed a hybrid phenotype during the disease, where epithelial features were associated with the presence of ≥ 5 CTCs/7.5 mL of blood, while high relative expression of the gene
MYCL
was observed preferentially in the set of samples with < 5 CTCs/7.5 mL of blood. At baseline, patients whose CTCs had stem or hybrid features showed a later progression. After 1 cycle of docetaxel, high relative expression of
ZEB1
indicated worse outcome, while
KRT19
and
KLK3
high expression could predisposed the patients to a worse prognosis at clinical progression. In the present work we describe biomarkers with clinical relevance for the prediction of early response or resistance in castration-resistant prostate cancer patients. Besides, we question the utility of targeted isolated CTCs and the use of a limited number of markers to define the CTCs population.
Abstract
Introduction: Cancer is a rapidly evolving, multifactorial disease that accumulates numerous genetic and epigenetic alterations. This results in phenotypic and molecular heterogeneity within ...the tumor, the complexity of which is further amplified through specific interactions between cancer cells and the tumor microenvironment. In this context, cancer may be perceived as an “ecomolecular” disease that involves cooperation between several neoplastic clones and their interactions with other cell types and secreted factors present in the microenvironment. Cancer is therefore analogous to complex ecosystems such as microbial consortia. We are currently aiming at dissecting the molecular mechanisms underlying the cooperation between different clones.
Methods: We produced clonal cell lines derived from the MDA-MB-231 breast cancer cell line, using UbC-StarTrack system, which allows tracking of multiple clones by color. The characteristics of each clone were determined by measuring migration, proliferation and metabolic activity. In vivo analysis by orthotopic breast injection and intravascular tail vein injection was performed to assess the tumorigenic capacity of these clones. We studied potential collaborations between clones by determining the influence of secreted factors on growth rate of different clones by medium complementation with supernatant or exosomes from different clones. In vivo, we used zebrafish as a model system to study the migration of individually or co-injected clones.
Results: Characterization of these clones in vitro revealed clear differences in proliferation, cell metabolic activity and morphology among them. In vivo, all the individually injected clones were able to form tumors but the growth rates differed among them. Injection of an equal mix of clones led to the formation of tumors where some clones displayed a growth or survival advantage. In vitro the complementation of growth medium with medium from other clone increased the proliferation rate of the other clones. Co-injection of clones in zebrafish increased the efficiency of migration.
Conclusions: These results confirm that even in stable cell lines heterogeneity is present. Malignant properties were enhanced when some clones were combined or treated with exosomes or medium from other clones. These results clearly support our hypothesis that tumor clones cooperate in cancer progression and that this cooperation is mediated by secreted factors. Finally, we are performing expression arrays of mRNA, microRNas and lncRNAs and determining epigenetic state by methylation arrays, in order to identify potential factors that are differentially expressed among clones and are therefore bona fide candidates for clonal cooperation promoting factors. It is anticipated that this knowledge will facilitate the design of new and more effective therapeutic approaches that are directed to the tumor ecosystem as a sum of different clones.
Citation Format: Ana Martín-Pardillos, Angeles Valls-Chiva, Eva Bejar Serrano, Roberto Piñeiro Cid, Pablo Hurtado Blanco, Angel Días-Lagares, María Magdalena Castro, Juan Antonio Juan Antonio Cámara Serrano, Santiago Ramon y Cajal. Clonal cooperation in cancer progression: A new paradigm in cancer 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 2183.
Cancer of Unknown Primary (CUP) is a metastatic cancer with confirmed histology of which the primary origin is unknown after to work up initial evaluation through a pathological clinical study, the ...analytical and imaging study.
The diagnostic process includes the early obtaining of quality biopsy material. It includes its histological and immunohistochemical analysis: a study to determine the tumor line, an analysis of cytokeratins and a large battery of antibodies to confirm the specific origin of each possible tumor type.
The development of molecular platforms has allowed improving the diagnosis of CUP, increasing the number of patients who can benefit from treatment with specific therapy, significantly increasing the survival and reducing the toxicity. However, the updated guidelines (NICE, ESMO, NCCN) emphasize that the impact on the clinical benefit of the specific treatment according to the results of the molecular platforms is still controversial.
El Cáncer de Origen Desconocido (COD) es un cáncer metastásico con histología confirmada del cual se desconoce el origen primario después de realizar un
estudio diagnóstico inicial mediante el estudio clínico patológico, el estudio analítico y de imagen.
El estudio diagnóstico incluye la obtención precoz de material de biopsia de calidad. Incluye su análisis histológico e inmunohistoquímico: un estudio para
determinar la estirpe tumoral, análisis de citoqueratinas y una batería amplia de anticuerpos para confirmar el origen específico.
El desarrollo de plataformas moleculares ha mejorado su diagnóstico, incrementando el número de pacientes que se benefician del tratamiento con
terapia específica, aumentando su supervivencia y reduciendo la toxicidad. Sin embargo, las guías actualizadas (NICE, ESMO, NCCN) destacan que el impacto
en el beneficio clínico del tratamiento específico según los resultados de las plataformas moleculares es todavía controvertido.
Antecedentes: El cáncer de células pequeñas (CPCP) representa el 13-15% del total de neoplasias primarias de pulmón. Se caracteriza por su rapidez en la tasa de crecimiento y en el desarrollo de ...metástasis a distancia. Objetivos: Orientar y estandarizar el tratamiento del CPCP, enfermedad limitada, en México, basado en evidencia clínica nacional e internacional. Material y métodos: Este documento se desarrolló como una colaboración entre el Instituto Nacional de Cancerología y la Sociedad Mexicana de Oncología en cumplimiento con estándares internacionales. Se integró un grupo conformado por oncólogos médicos, cirujanos oncólogos, cirujanos de tórax, radio-oncólogos y metodólogos con experiencia en revisiones sistemáticas de la literatura y guías de práctica clínica. Resultados: Se consensuaron, por el método Delphi y en reuniones a distancia, las recomendaciones en CPCP enfermedad limitada, producto de las preguntas de trabajo. Se identificó y evaluó críticamente la evidencia científica que responde a cada una de dichas preguntas clínicas, antes de incorporarla a la guía. Conclusión: Esta guía proporciona recomendaciones clínicas para el manejo de la enfermedad limitada del CPCP y durante el proceso de toma de decisiones de los clínicos involucrados con su manejo en nuestro país para mejorar la calidad de la atención clínica en estos pacientes.
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