Person:
Peña Fernández, Laura Luisa

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First Name
Laura Luisa
Last Name
Peña Fernández
URI
https://hdl.handle.net/20.500.14352/80626
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Medicina y Cirugía Animal
Area
Medicina y Cirugía Animal
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    Histopathological and immunohistochemical findings in lymphoid tissues of the endangered Iberian lynx (Lynx pardinus)
    (Comparative Immunology, Microbiology and Infectious Diseases, 2006) Peña Fernández, Laura Luisa; García Palencia, María Del Pilar; Jiménez Martínez, María De Los Ángeles; Benito, Alberto; Pérez Alenza, María De Los Dolores; Sánchez Maldonado, María Belén
    The Iberian lynx (Lynx pardinus) is the most threatened wild feline in the world. Little is known about the diseases and pathology that affect this animal. The aim of this study was to evaluate the histopathological status of the peripheral lymphoid tissues and thymus of Iberian lynxes necropsied between 1998 and 2003. Seventeen animals including females (n=8) and males (n=9), age range of 10 months to 16 years, with different causes of death were histopathologically and immunohistochemically (anti-CD3, CD79, MAC387, CD68) studied. Feline immunosuppressive virus laboratorial tests were negative. Five individuals presented neoplasia and/or tuberculosis. All animals presented some degree of both B and T cells depletion in peripheral lymphoid tissues and follicular hyalinosis in the center of depleted follicles. A viral origin of the lymphoid depletion is postulated although other causes (inbreeding, stress, toxic) are not ruled out. The loss of the effectiveness of the immune system increases the vulnerability of the critically endangered Iberian lynx to pathogens
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    Establishment and Characterization of a New Cell Line of Canine Inflammatory Mammary Cancer: IPC-366
    (PLoS ONE, 2015) Cáceres Ramos, Sara Cristina; Peña Fernández, Laura Luisa; Andrés Gamazo, Paloma Jimena De; Illera Del Portal, Josefina María; Lopez, Mirtha S.; Woodward, Wendy A.; Reuben, James M.; Illera Del Portal, Juan Carlos
    Canine inflammatory mammary cancer (IMC) shares epidemiologic, histopathological and clinical characteristics with the disease in humans and has been proposed as a natural model for human inflammatory breast cancer (IBC). The aim of this study was to characterize a new cell line from IMC (IPC-366) for the comparative study of both IMC and IBC. Tumors cells from a female dog with clinical IMC were collected. The cells were grown under adherent conditions. The growth, cytological, ultrastructural and immunohistochemical (IHC) characteristics of IPC-366 were evaluated. Ten female Balb/SCID mice were inoculated with IPC-366 cells to assess their tumorigenicity and metastatic potential. Chromosome aberration test and Karyotype revealed the presence of structural aberration, numerical and neutral rearrangements, demonstrating a chromosomal instability. Microscopic examination of tumor revealed an epithelial morphology with marked anysocytosis. Cytological and histological examination of smears and ultrathin sections by electron microscopy revealed that IPC-366 is formed by highly malignant large round or polygonal cells characterized by marked atypia and prominent nucleoli and frequent multinucleated cells. Some cells had cytoplasmic empty spaces covered by cytoplasmic membrane resembling capillary endothelial cells, a phenomenon that has been related to s vasculogenic mimicry. IHC characterization of IPC-366 was basal-like: epithelial cells (AE1/AE3+, CK14+, vimentin+, actin-, p63-, ER-, PR-, HER-2, E-cadherin, overexpressed COX-2 and high Ki-67 proliferation index (87.15 %). At 2 weeks after inoculating the IPC-366 cells, a tumor mass was found in 100 % of mice. At 4 weeks metastases in lung and lymph nodes were found. Xenograph tumors maintained the original IHC characteristics of the female dog tumor. In summary, the cell line IPC-366 is a fast growing malignant triple negative cell line model of inflammatory mammary carcinoma that can be used for the comparative study of both IMC and IBC.
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    Steroids and receptors in canine mammary cancer
    (Steroids, 2006) Illera Del Portal, Juan Carlos; Pérez Alenza, María De Los Dolores; Nieto Ruiz De Zárate, Ana Isabel; Jiménez, María; Silván Granado, Gema; Dunner Boxberger, Helene Susana; Peña Fernández, Laura Luisa
    The aims of this study were to investigate the serum and tissue content of androgens and estrogens in canine inflammatory mammary carcinomas (IMC) as well as in non-inflammatory malignant mammary tumors (MMT), and assessed the immunoexpression of estrogen and androgen receptors using immunohistochemistry. Profiles for the androgens dehydroepiandrosterone (DHEA), androstenedione (A4), and testosterone (T), and for the estrogens 17β estradiol (E2) and estrone-sulphate (SO4E1) were measured both in tissue homogenates and in serum of MMT and IMC by EIA techniques in 42 non-inflammatory malignant mammary tumors (MMT) and in 14 inflammatory mammary carcinomas (IMC), prospectively collected from 56 female dogs. Androgen receptor (AR) and estrogen receptor alpha (ERα) and beta (ERβ) expression was studied using immunohistochemistry (strepavidin–biotin-peroxidase method) in samples of 32 MMT and 14 IMC, and counted by a computer image analyzer. IMC serum and tissue levels of androgens were significantly higher than MMT levels. Tissue content of estrogens was also significantly higher in IMC than in MMT. Serum values of SO4E1 were significantly higher in IMC, but serum levels of E2 were significantly lower in IMC compared to MMT cases. Medium-high androgen receptor intensity was observed in 64.28% of IMC and 40.62% of MMT. No important differences were found between ERα expression in IMC (100% negative) and MMT (90% negative). ERβ and AR were intensely expressed in highly malignant inflammatory mammary carcinoma cells. To our knowledge, this is the first report relative to AR immunohistochemistry in canine mammary cancer and to estrogens or androgens in serum of dogs with benign or malignant mammary tumors.
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    Transcriptomics of Canine Inflammatory Mammary Cancer Treated with Empty Cowpea Mosaic Virus Implicates Neutrophils in Anti-Tumor Immunity
    (International Journal of Molecular Sciences, 2023) Barreno San Antolín, Lucía; Sevane Fernández, Natalia; Valdivia Lara, Edgar Guillermo; Alonso Miguel, Daniel; Suárez Redondo, María; Alonso Díez, Ángela; Fiering, Steven; Beiss, Veronique; Steinmetz, Nicole F.; Pérez Alenza, María De Los Dolores; Peña Fernández, Laura Luisa
    Canine inflammatory mammary cancer (IMC) is a highly aggressive and lethal cancer in dogs serving as a valuable animal model for its human counterpart, inflammatory breast cancer (IBC), both lacking effective therapies. Intratumoral immunotherapy (IT-IT) with empty cowpea mosaic virus (eCPMV) nanoparticles has shown promising results, demonstrating a reduction in tumor size, longer survival rates, and improved quality of life. This study compares the transcriptomic profiles of tumor samples from female dogs with IMC receiving eCPMV IT-IT and medical therapy (MT) versus MT alone. Transcriptomic analyses, gene expression profiles, signaling pathways, and cell type profiling of immune cell populations in samples from four eCPMV-treated dogs with IMC and four dogs with IMC treated with MT were evaluated using NanoString Technologies using a canine immune-oncology panel. Comparative analyses revealed 34 differentially expressed genes between treated and untreated samples. Five genes (CXCL8, S100A9, CCL20, IL6, and PTGS2) involved in neutrophil recruitment and activation were upregulated in the treated samples, linked to the IL17-signaling pathway. Cell type profiling showed a significant increase in neutrophil populations in the tumor microenvironment after eCPMV treatment. These findings highlight the role of neutrophils in the anti-tumor response mediated by eCPMV IT-IT and suggest eCPMV as a novel therapeutic approach for IBC/IMC.
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    Project number: PIMCD155/23-24
    Elaboración de un atlas fotográfico en formato digital y físico de órganos sanos como complemento a la docencia de Anatomía Patológica Especial y Rotatorio Clínico
    (2024) Andrés Gamazo, Paloma Jimena De; García Fernández, Rosa Ana; Jiménez Martínez, María De Los Ángeles; García Palencia, María Del Pilar; Sánchez Maldonado, María Belén; Peña Fernández, Laura Luisa; González Huecas, Marta; Tabanera De Lucio, Enrique; Barreno San Antolín, Lucía; Valdivia Lara, Edgar Guillermo; Sánchez Pérez, María De Los Ángeles; Andrés Gamazo, Paloma Jimena De
    El proyecto se centró en crear un atlas fotográfico digital y físico de órganos sanos para estudiantes de Veterinaria, con el objetivo de mejorar la identificación de lesiones durante las necropsias y fomentar el aprendizaje autónomo. A raíz de la pandemia de COVID-19, se incorporaron tecnologías digitales en la docencia, lo que subrayó la necesidad de herramientas didácticas innovadoras en el Grado en Veterinaria, que tiene una elevada carga práctica. Para realizar el atlas se recopilaron y clasificaron imágenes de órganos sanos de diferentes fuentes, se creó un espacio en el Campus Virtual para mostrar el atlas en su versión digital, y paralelamente las imágenes se imprimieron y plastificaron para su uso en la sala de necropsias. Finalmente, se realizó una evaluación de la eficacia del atlas en ambos formatos mediante encuestas anónimas a los estudiantes. Los resultados preliminares fueron difundidos internacionalmente en el VII Congreso Español y I Hispano-Portugués de Docencia Veterinaria organizado por la Asociación Española de Docentes Veterinarios (VetDoc). Los resúmenes de los trabajos, además de en el libro de resúmenes del congreso, serán publicados en la revista Frontiers in Veterinary Science. La recopilación de imágenes resultó desafiante, pero se logró recopilar una suficiente cantidad de imágenes de las especies canina, felina, equina, porcina, bovina y de pequeños rumiantes. La implementación en el Campus Virtual facilitó el acceso continuo al recurso, mientras que la versión física fue esencial para las prácticas en la sala de necropsias. La evaluación del proyecto, a través de encuestas anónimas, mostró que los estudiantes apreciaron y utilizaron el atlas en ambos formatos, destacando su utilidad para otras asignaturas y su efectividad en mejorar el proceso de enseñanza y aprendizaje. La preferencia general fue por el formato digital, aunque el físico también fue muy valorado en las prácticas. En conclusión, el proyecto no solo mejoró la enseñanza de Anatomía Patológica en Veterinaria, sino que también demostró la validez de integrar tecnologías digitales en la educación práctica. La metodología y los resultados pueden servir como referencia para futuros proyectos educativos, contribuyendo al avance de la educación veterinaria.
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    Project number: 278
    Diseño y desarrollo de herramientas para el auto-aprendizaje de Anatomía Patológica General Veterinaria
    (2016) Peña Fernández, Laura Luisa; González Huecas, Marta; Tabanera De Lucio, Enrique; García Fernández, Rosa Ana; Pizarro Díaz, Manuel
    Este proyecto es la continuación de otro anterior de innovación docente: PIMCD 238/2014 “Implantación de un sistema de telepatología digital para la enseñanza práctica de Anatomía Patológica General (asignatura de 2º curso del Grado en Veterinaria)”. Hemos elaborado una colección completa de preparaciones digitalizadas para todas las prácticas de histopatología de APG. Con la colección completa de preparaciones histopatológicas digitalizadas, se han realizado también guías específicas de auto-ayuda para una mejor comprensión y observación de las mismas por parte del alumno, favoreciendo el auto-aprendizaje individualizado y en grupo. Así mismo, hemos realizado un módulo de auto-aprendizaje teórico-práctico en inglés para que el alumno auto-evalúe sus conocimientos de partes de la asignatura y los conocimientos transversales de otras asignaturas del grado en veterinaria. Como complemento se ha producido un juego de “pasapalabra” con contenidos de Anatomía Patológica, con el fin de estimular el aprendizaje mediante el juego. Ambos sistemas se encuentran en el campus virtual UCM para su utilización por los alumnos de veterinaria. Al finalizar el curso se enlazarán en la página web del Dpto.
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    Canine cell line, IPC‐366, as a good model for the study of inflammatory breast cancer
    (Veterinary and Comparative Oncology, 2016) Cáceres Ramos, Sara Cristina; Peña Fernández, Laura Luisa; Lacerda, Lara; Illera Del Portal, Josefina María; Andrés Gamazo, Paloma Jimena De; Larson, Richard; Gao, Hui; Debeb, Bisrat; Woodward, Wendy; Reuben, James; Illera Del Portal, Juan Carlos
    Inflammatory breast cancer (IBC) is an aggressive type of cancer with poor survival in women. Inflammatory mammary cancer (IMC) in dogs is very similar to human IBC and it has been proposed as a good surrogate model for study the human disease. The aim was to determine if IPC-366 shared characteristics with the IBC cell line SUM149. The comparison was conducted in terms of ability to grow (adherent and nonadherent conditions), stem cell markers expression using flow cytometry, protein production using western blot and tumorigenic capacity. Our results revealed that both are capable of forming long-term mammospheres with a grape-like morphology. Adherent and nonadherent cultures exhibited fast growth in vivo. Stem cell markers expressions showed that IPC-366 and SUM149 in adherent and nonadherent conditions has mesenchymal-like characteristics, E-cadherin and N-cadherin, was higher in adherent than in nonadherent cultures. Therefore, this study determines that both cell lines are similar and IPC-366 is a good model for the human and canine disease.
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    Aqueous dispersions of oleic acid nanodroplets for thymol encapsulation
    (Colloids and Surfaces A, 2024) Paula Gutiérrez-González; Peña Fernández, Laura Luisa; Alejandro Lucia; Ortega Gómez, Francisco; González Rubio, Ramón; Guzmán Solís, Eduardo
    This study focuses on the use of oleic acid to disperse thymol as nanodroplets, stabilized by a mixture of alkylpolyglucoside and lecithin, in an aqueous environment. This approach aims to develop innovative platforms for the encapsulation and release of poorly water-soluble molecules such as thymol, useful for drug delivery and insecticide systems. The results highlight the critical role of controlling the content and concentration of the oil phase (thymol-oleic acid mixture) in achieving optimal thymol dispersion and nanodispersion stability. The interplay between the ability of oleic acid to inhibit thymol crystallization and the maximum dispersible oil amount is crucial. It affects the dispersion of thymol within the nanodroplets and influences coalescence and Ostwald ripening phenomena. The balance between oleic acid and thymol content is key: while oleic acid stabilizes dispersions, higher thymol content increases droplet size, potentially triggering destabilization. The uneven distribution of thymol within the droplets, revealed by fluorescence spectroscopy, suggests that up to three different chemical environments exist. This investigation may pave the way for the development of efficient platforms to improve access to biologically relevant, poorly soluble molecules.
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    Neoadjuvant Intratumoral Immunotherapy with Cowpea Mosaic Virus Induces Local and Systemic Antitumor Efficacy in Canine Mammary Cancer Patients
    (Cells, 2023) Valdivia Lara, Edgar Guillermo; Alonso Miguel, Daniel; Pérez Alenza, María De Los Dolores; Zimmermann, Anna Barbara Emilia; Schaafsma, Evelien; Kolling IV, Fred W.; Barreno San Antolín, Lucía; Alonso Díez, Ángela; Beiss, Veronique; Affonso de Oliveira, Jessica Fernanda; Suárez Redondo, María; Fiering, Steven; Steinmetz, Nicole F.; Vom Berg, Johannes; Peña Fernández, Laura Luisa; Arias Pulido, Hugo
    The lack of optimal models to evaluate novel agents is delaying the development of effective immunotherapies against human breast cancer (BC). In this prospective open label study, we applied neoadjuvant intratumoral immunotherapy with empty cowpea mosaic virus-like particles (eCPMV) to 11 companion dogs diagnosed with canine mammary cancer (CMC), a spontaneous tumor resembling human BC. We found that two neoadjuvant intratumoral eCPMV injections resulted in tumor reduction in injected tumors in all patients and in noninjected tumors located in the ipsilateral and contralateral mammary chains of injected dogs. Tumor reduction was independent of clinical stage, tumor size, histopathologic grade, and tumor molecular subtype. RNA-seq-based analysis of injected tumors indicated a decrease in DNA replication activity and an increase in activated dendritic cell infiltration in the tumor microenvironment. Immunohistochemistry analysis demonstrated significant intratumoral increases in neutrophils, T and B lymphocytes, and plasma cells. eCPMV intratumoral immunotherapy demonstrated antitumor efficacy without any adverse effects. This novel immunotherapy has the potential for improving outcomes for human BC patients.
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    In vitro and in vivo effect of flutamide on steroid hormone secretion in canine and human inflammatory breast cancer cell lines
    (Veterinary and Comparative Oncology, 2017) Cáceres Ramos, Sara Cristina; Monsalve, Beatriz; Peña Fernández, Laura Luisa; Andrés Gamazo, Paloma Jimena De; Alonso‐Diez, Ángela; Illera Del Portal, Josefina María; Woodward, Wendy; Reuben, James; Silván Granado, Gema; Illera Del Portal, Juan Carlos
    The aim was to study the effects of flutamide on cell proliferation, in vivo tumour growth andsteroid production in canine and human IBC cell lines. IPC-366 and SUM149 cell cultures wereexposed to flutamide concentrations for 72 hours. Additionally, IPC-366 and SUM149 xeno-transplanted mice were treated subcutaneously with flutamide 3 times a week for 2 weeks.Steroid hormones determination in culture media, serum and tumour homogenates (pregneno-lone, progesterone, androstenedione, testosterone, dihydrotestosterone, 17β-oestradiol andoestrone sulphate) were assayed by EIA. in vitro cell proliferation percentages showed adecrease in all flutamide dosages in IPC-366 and SUM149. in vivo flutamide reduced tumoursize by 55% to 65%, and metastasis rates decreased. In treated groups, androgen levels in cul-ture media, serum and tumour homogenates were increased as oestrogen levels decreased. These results suggest that flutamide treatment inhibits cell proliferation and promotes tumourreduction by increasing androgen levels and also support future therapy approaches