dianas 7(1) > Sánchez-Martín etal

dianas | Vol 7 Num 1 | marzo 2018 | e201803p09

Bases moleculares de la actividad anti-tumoral de nuevos agentes terapéuticos derivados de la hispanolona.

Vanesa Sánchez Martín^a, Sonsoles Hortelano Blanco, Beatriz de las Heras Polo

Instituto de Salud Carlos III / Universidad Complutense de Madrid.

a. vanesa_sanchez_92@hotmail.com

III Congreso de Señalización Celular, SECUAH 2018.
20-22 de marzo, 2018. Universidad de Alcalá. Alcalá de Henares, Madrid. España.
Sesión de paneles

Resumen

Los productos naturales desempeñan un papel fundamental en el descubrimiento de nuevos fármacos, particularmente evidente en áreas como el cáncer, donde alrededor del 60% de los agentes terapéuticos utilizados son de origen natural. Los terpenos son un grupo químico de amplia distribución en la naturaleza a los que se les han atribuido actividades biológicas diversas: antiinflamatoria, antiviral, antiproliferativa y antibacteriana, entre otras. En este proyecto queremos evaluar el potencial anti-tumoral de un diterpeno de tipo labdano derivado de la hispanolona, el α-hispanolol, en células de glioblastoma, uno de los tumores cerebrales más agresivos a día de hoy y para el que todavía no existen terapias eficaces. Nuestros resultados muestran que el α-hispanolol produce cambios morfológicos y disminuye la viabilidad celular de las líneas celulares U87 y U373 de manera dosis- y tiempo-dependiente. Además, el compuesto induce apoptosis y arresta el ciclo celular en la fase G0/G1, acumulándose las células en la fase sub-G0. El α-hispanolol aumenta la actividad de las caspasas y la expresión de proteínas pro-apoptóticas (Bax y Bid) e inhibe la expresión de proteínas anti-apoptóticas (Bcl-2 y Bcl-xl). Además, el compuesto presenta un efecto inhibitorio de la migración y de la invasión, disminuyendo la actividad de las MMPs. Como conclusión, nuestros datos sugieren que el α-hispanolol podría tener potencial terapéutico en el tratamiento de los glioblastomas.

dianas 7(1) > Yllera-García etal

dianas | Vol 7 Num 1 | marzo 2018 | e201803p08

Exosomes as new tools for searching relevant proteins in colorectal cancer metastasis.

Beatriz Yllera García, Vivian de los Rios Benítez, José Ignacio Casal Álvarez

Centro de Investigaciones Biológicas, CSIC.

a. beatriz.yllera@gmail.com

III Congreso de Señalización Celular, SECUAH 2018.
20-22 de marzo, 2018. Universidad de Alcalá. Alcalá de Henares, Madrid. España.
Sesión de paneles

Resumen

Exosomes are extracellular vesicles with a wide range of functions, including metastasis seeding, in most types of cancer. The aim of this study was to find proteins critical for colorectal cancer metastasis. A non-metastatic murine model of colorectal cancer was compared with the metastatic mouse cell line CT-26 to investigate the exosomal protein content. To optimize the recovery of exosomes, cells were incubated in serum-free DMEM for 48h. Colon tissues were incubated for 2, 4 and 8h in serum-free DMEM. The exosomes were isolated by ultracentrifugation and lysated with different buffers: 7M urea/2M thiourea, methanol-chloroform and RIPA buffer. Exosomal lysates were analyzed in the Q-Exactive using a gradient: 5–35% (90min), 35%-100% (4min) and 100% (8min) ACN with 0.1% formic acid. Mass spectra were acquired using a “top 15” method and searched against SwissProt using Sequest HT search engine through Proteome Discoverer. Identified peptides were filtered using Percolator with a q-value threshold of 0.01 at a FDR of 1%. The largest amount of mouse exosomes from colon was recovered at 2h. The 7M urea / 2M thiourea buffer was the most effective lysis method, with 35% more exosomal proteins detected by mass spectrometry. The identified proteins correlated with: remodelling of the ECM, organotropism, angiogenesis and chemo-attraction, EMT, cellular metabolism, etc. Significant differences were observed between metastatic and non-metastatic exosomes. Proteins of metastatic exosomes are largely related to the interaction and modulation of the extracellular matrix and cell migration, including α3 and α5 integrins, in contrast to the non-metastatic exosomes. Laminins α5, β1 and ɣ1, which form laminin-511, are major candidates for further studies. Methods for extracting and lysing exosomes from mouse colon and metastatic cells were optimised. A preliminary protein profile involved in metastasis has been obtained and will be further analysed using functional experiments. Keywords: exosomes, metastasis, colorectal cancer, extracellular matrix, mouse models.

dianas 7(1) > Lázaro etal

dianas | Vol 7 Num 1 | marzo 2018 | e201803p05

Lung adenocarcinoma and sarcomatoid carcinoma mouse model.

Sara Lázaro, Miriam Pérez, Ana Belén Enguita, Laura Gómez, Jesús M. Paramio, Mirentxu Santos

CIEMAT.

a. sara.lazaro@ciemat.es

III Congreso de Señalización Celular, SECUAH 2017.
20-22 de marzo, 2018. Universidad de Alcalá. Alcalá de Henares, Madrid. España.
Sesión

dianas 7(1) > Ruiz-López etal

dianas | Vol 7 Num 1 | marzo 2018 | e201803p04

Virtual Biopsy: development of non-invasive immunotargeted imaging agents for the diagnosis of glioblastoma.

Eduardo Ruiz-López, Ruth González-Gómez, Beatriz Torres-Herrero, Sara Naya-Forcano, Natalia Magro, Héctor Tejero, Fátima Al-Shahrour, Luis Oriol, Miguel Ángel Morcillo, Alberto Jiménez Schuhmacher

Instituto de Investigación Sanitaria de Aragón (IIS Aragón).

a. educalata21@gmail.com

III Congreso de Señalización Celular, SECUAH 2017.
20-22 de marzo, 2018. Universidad de Alcalá. Alcalá de Henares, Madrid. España.
Sesión

Resumen

Glioblastoma (GBM) is the most common and aggressive brain tumor. Patient survival has improved marginally during the past 3 decades, averaging 14.6 months with only 3-5% of patients surviving more than 3 years. Current diagnosis by Magnetic Resonance Imaging (MRI) provides morphological information, which is sometimes inaccurate, and the final diagnosis of GBM often requires a brain biopsy. These biopsies may not depict the high intratumoral heterogeneity present in GBM and may favor its recurrence. These disadvantages highlight the importance of looking for other molecular imaging based technologies that allow efficient and safe diagnosis. Positron Emission Tomography (PET) is an alternative but unfortunately, the most widely used tracer 18F-Fluorodeoxyglucose (18F-FDG) is ineffective in GBM due to the high consumption of glucose by the brain. Other radiotracers for GBM, mostly interrogating the metabolism, are under evaluation. An innovative option is termed immunotargeted imaging. This approach combines the target selectivity and specificity of antibodies and engineered fragments against a given tumor cell surface marker with the main characteristics (high spatial resolution, sensitivity, quantitative capabilities) of imaging techniques. ImmunoPET could be considered a virtual biopsy, comparable to conduct an in vivo, integrated, quantifiable, 3D, full body immunohistochemistry allowing the diagnosis and monitoring of patients over time in a non-invasive manner. Applying bioinformatics on patient datasets, we have identified ATP Binding Cassette Subfamily C Member 3 (ABCC3) as a molecular target for the development of an immunoPET agent for GBM, due to the high levels of ABCC3 protein expression in GBM and negligable levels in normal brain. Importantly, ABCC3 is located in the plasma membrane and the increase of its expression is associated with tumor grade, poor survival and impaired response to the most currently used chemotherapy. We propose to develope a novel immunoPET agent against ABCC3. First, we will validate ABCC3 as a suitable target for immunoimaging using a murine monoclonal antibody labeled with 89Zirconium in xenograft mice and avatar/tumorgraft models. For further clinical development, we will need to reduce the radioactivity exposure and optimize the blood brain barrier permeability of the tracer. To this end, we will generate different antibody fragments with faster clearance and pharmacokinetics. As matching of the physical half-life of the positron-emitting radionuclide with the biological half-life of the antibody or fragment is crucial for immunoPET, these smaller derivatives will be labeled with PET isotopes of shorter half-lives, such as 68Galium, which advantageously can be produced in a generator rather than a cyclotron. Finally, we will use click chemistry by biorthogonal reactions in vivo in order to match other antibodies/fragments with tracers and thereby diminishing the exposure to radioactivity and ensuring a better signal-to-noise ratio. Click chemistry also allows to label multiple imaging tracers (MRI-tracers such as (Gd)-complexes or SPIO nanoparticles) to the same pretargeted molecule as well as multi-modal, multifunctional imaging and teragnostics. The successful development of these tracers has the potential to improve diagnoses, and may have a major impact on the survival of patients with GBM.