Programa y Libro de Abstracts del VI Simposio de Terapias Avanzadas y Tecnologías Biomédicas del ibs.GRANADA

INSCRIPCIONES COMITÉ ORGANIZADOR INFORMACIÓN GENERAL El Área de Terapias Avanzadas y Tecnologías Biomédicas del instituto de Investigación Biosanitaria ibs.GRANADA organiza el VI SIMPOSIO DE TERAPIAS AVANZADAS Y TECNOLOGÍAS BIOMÉDICAS. Tendrá lugar el viernes 28 de noviembre de 2025 en el Salón de Actos la Facultad de Medicina de la Universidad de Granada. OBJETIVOS El objetivo de este simposio es presentar las principales líneas de investigación llevadas a cabo en el área de las Terapias Avanzadas y las Tecnologías Biomédicas por grupos del ibs.GRANADA y grupos afines a estas áreas de conocimiento. El simposio supone una oportunidad para que investigadores y profesionales sanitarios puedan interaccionar y establecer nuevas líneas de colaboración y sinergias en el campo de las Terapias Avanzadas y las Tecnologías Biomédicas. Este simposio es además una oportunidad para difundir estos trabajos y proyectos al resto de profesionales sanitarios, así como al público en general. La asistencia es libre y gratuita con previa inscripción a través del siguiente enlace: https://www.ibsgranada.es/inscripcion-simposio-terapias-avanzadas-tecnologias-biosanitarias-ibsgranada/ El plazo límite pare realizar la inscripción es el 27 de noviembre de 2025. Jesús Chato Astrain. Prof. Departamento de Histología, Facultad de Medicina, Universidad de Granada. TEC03-Ingeniería Tisular. Óscar Darío García García. Prof. Departamento de Histología, Facultad de Medicina, Universidad de Granada. TEC03-Ingeniería Tisular. Francisco Javier Molina Estévez. Investigador del Departamento de Biología Celular. TEC24Grupo de Investigación en Hematología y Terapia Génica: HemaTerGe. COMITÉ CIENTÍFICO Araceli Aguilar González David Sánchez Porras Fabiola Bermejo Casares Fernando Campos Sanchez Francisco Martin Molina Julia Corral Sarasa María Tristán Manzano Miguel Alaminos Mingorance Miguel Angle Martín Piedra Miguel Etayo Escanilla Olimpia Ortiz Arrabal Paula Ávila Fernández El programa y libro de Abstracts del VI Simposio de Terapias Avanzadas y Tecnologías Biomédicas del ibs.GRANADA se encuentra públicamente disponible en la web del ibs.GRANADA y el en repositorio europeo Zenodo (https://zenodo.org/). 08:30-09:00h Reunión de coordinación de los investigadores responsables del Área de Terapias Avanzadas y Tecnologías Biomédicas 09:00-09:20h Inauguración del Simposio • Dr. José Juan Jiménez Moleón. Decano de la Facultad de Medicina. Universidad de Granada • Dr. Indalecio Sánchez-Montesinos García. Delegado territorial de la Consejería de Salud y Consumo de la Junta de Andalucía • Dra. María Ángeles García Rescalvo. Directora Gerente del Universitario Virgen de las Nieves • Dr. Manuel Enrique Reyes Nadal. Director Gerente del Hospital Universitario Clínico San Cecilio • Dra. María José Sánchez Pérez. Directora Científica de ibs.GRANADA • Dr. Óscar Darío García García. Organizador del VI Simposio de Terapias Avanzadas y Tecnologías Biomédicas. 09:20-09:40h Conferencia Invitada: PV-Skin: Un sustituto de piel con cuatro tipos celulares para grandes quemados Dra. Rocío García de la Cruz Valencia Translational Research Manager, Kinderspital Zürich, Suiza 09:40-10:55h Mesa 1: Terapias avanzadas: Ingeniería Tisular • Exosomas derivados de células madre mesenquimales en regeneración tisular. D. Víctor Javier Costela Ruz. Investigador del grupo TEC17 – BIOTEJSALUD ibs.GRANADA • Comunicación mediada por vesículas extracelulares entre plaquetas y células tumorales: rol en la carcinogénesis y trombosis en cáncer de páncreas (Estudio placetro) D. Antonio Palomeque Jiménez. Investigador del grupo TEC13-Cirugía Avanzada ibs.GRANADA PROGRAMA • Skin Cancer Model platform for drug testing Dña. Paula Ávila Fernández. Investigadora del grupo TEC03 – Ingeniería Tisular ibs.GRANADA • In vitro reconstituted ECMs for personalized tumor modelling Dña. Julia López de Andrés. Investigadora del grupo Grupo: TEC16 – Terapias Avanzadas: Diferenciación, Regeneración y Cáncer ibs.GRANADA • Diseño y Caracterización de Hidrogeles Multisensibles basados en Quitosano, Agarosa y Partículas Magnéticas D. Modesto Torcuato López López. Investigador del grupo Grupo TEC05-Física de Interfases y Sistemas Coloidales ibs.GRANADA 10:5511:40 h Mesa 2: Terapias Avanzadas: Terapia Génica y Farmacogenética • Advanced gene editing applied to safer gene therapy and novel Pompe disease models D. Francisco Javier Molina Estévez. Investigador del grupo Grupo: TEC24-Grupo de Investigación en Hematología y Terapia Génica: HemaTerGe ibs.GRANADA • Dual-CRISPR Systems: Advancing Genome Editing and Diagnostics Dña. Araceli Aguilar González. Investigadora del grupo TECE18 – NANOCHEMBIO ibs.GRANADA • Práctica Segura en el Uso de Medicamentos: Integración de la Farmacogenética con GenHUSC App y Desconstrucción de la Vacuna de ARNm-NPLs D. Xando Díaz Villamarín y Pilar Baena Álvarez. Investigadores del grupo TEC01 – Práctica segura en el uso de medicamentos ibs.GRANADA 11:40-12:20h Café y visita a pósteres 12:20-12:30h Conferencia Patrocinada: Innovación tecnológica en Biología Celular y Terapias Avanzadas D. Aitor González Granja Marketing Manager at Izasa by Palex 12:3013:30 h Mesa 3: Salud y Rehabilitación: Estrategias Clínicas y Tecnológicas • Asociación de cambios en la materia gris con los resultados funcionales y de calidad de vida en pacientes con ictus tras cirugía de la extremidad superior espástica. Resultados a seis meses de un ensayo aleatorizado Dña. Patricia Hurtado Olmo. Investigadora del grupo Grupo: TECE21 – Reparación, Regeneración y Sustitución Ósea ibs.GRANADA • Intervención multimodal en dolor lumbar crónico D. Víctor Segura-Jiménez. Investigador del grupo TECE20 – REHABILITA-T: Avances e Innovación en Rehabilitación y Promoción de la Salud ibs.GRANADA • Inflamación y riesgo cardiovascular en psoriasis e hidradenitis D. Manuel Sánchez Díaz. Investigador del grupo TECE19 – Dermatología Clínica y Traslacional ibs.GRANADA • Avances en enfermedad renal hereditaria D. Rafael J Esteban De la Rosa. Investigador del grupo TEC14 – Reproducción Humana y Enfermedades Hereditarias y Complejas ibs.GRANADA 13:30-14:15h Mesa 4: Imagen Médica y Modelado Computacional • PET/TC en inflamación vascular en hipercolesterolemia familiar Dña. María E. Bellón Guardia. Investigadora del grupo TEC15 – Medicina Nuclear y Molecular ibs.GRANADA • Modelado computacional del color dental Dña. María Navidad Tejada Casado. Investigadora del grupo TEC09 – Óptica de Biomateriales y Tejidos ibs.GRANADA • Algoritmo Cervisense TPTL para predicción de parto prematuro Dña. Paqui Molina. Investigadora del grupo TEC12 – Salud Materno Fetal y Elastografía ibs.GRANADA 14:15h Cierre de la jornada y conclusiones LIBRO DE COMUNICACIONES ÍNDICE DE COMUNICACIONES PRESENTADAS EN EL SIMPOSIO ABSTRACT INDEX Integración de la inteligencia artificial para fortalecer la precisión del diagnóstico histopatológico en cáncer de próstata 1 Adhesive performance of novel nanoparticle formulations doped with dexamethasone and doxycycline at the dentin–resin interface 2 Green Synthesis and Characterization of Icariin-Loaded Beeswax Solid Lipid Nanoparticles for Innovative Dental Therapies 3 Del carbón activado a las terapias dirigidas: materiales magneto-activos para liberación controlada y tratamiento localizado 4 Construcción y visualización de un espacio cromático dental difuso mediante espacios conceptuales: PyFCS 5 Does the chromatic ability of one-shaded resin-based composites limited by the surrounding? Preliminary results 6 Herramienta basada en inteligencia artificial para estimar el fracaso en la extracción testicular de espermatozoides en pacientes con azoospermia no obstructiva 7 Caracterización multiómica de células Sertoli adultas como base para entender la disfunción reproductiva masculina 8 Unveiling the immune landscape of Sertoli cell-only testes through single-cell transcriptomics in non-obstructive azoospermia 9 Single-cell transcriptomic profiling uncovers seminal cellular diversity in nonobstructive azoospermia 10 Genetic Evidence from Mendelian Randomization Supports a Causal Role of Immune-Mediated Diseases in Severe Male Infertility 11 Advancing CSC-Targeted Approaches in PDAC Using a Reproducible Preclinical Model of Liver Metastasis 12 Impact of prevalent KRAS mutations in PDAC resistance to FIRINOX 13 Acción de los exosomas derivados de células madre mesenquimales de médula ósea en la modulación de parámetros fibroblásticos asociados con la regeneración tisular 14 Exosomas derivados de células madre mesenquimales de médula ósea promueven la proliferación y migración de osteoblastos humanos 15 Potencial de los extractos fenólicos de alperujo en la modulación de la inflamación y la proliferación celular 16 Análisis comparativo de la Interacción entre dos CARs anti-CD19 mediante herramientas bioinformáticas. 17 Next-generation inducible promoters for safer and more specific CAR-T therapies 18 ALHAMBRA: A Novel Promoter for Activation-Dependent Expression in FourthGeneration CAR-T Cells 19 Exploring Dual-Guide RNA architectures to enhance CRISPR/Cas13-based diagnostics 20 Development of Hybrid Nanoparticles for Dual Activation Cancer Treatment via Magnetic and NIR Fields 21 Nanocatalytic Platforms and Bioorthogonal Prodrug Systems to Enhance CAR-T Cell Immunotherapy in Breast Cancer 22 Fluorescent Copper Nanocatalyst for Tracking Azide-Alkyne Cycloadditions in both solutions and cells 23 A Pd-modified porphyrinic MOF for combined bioorthogonal catalysis and photodynamic therapy 24 Beyond Standard Treatment: Can Exercise and Mindfulness Enhance Disability in Individuals with Chronic Primary Low Back Pain? The BACKFIT project 25 Melatonin Ameliorates Heart Failure by Improving Ventricular Remodeling and Ejection Fraction in Male ZDF Rats 26 Conexiones genéticas entre la depresión mayor y la obesidad: un enfoque desde la secuenciación de nueva generación 27 Dissecting the Molecular Basis of Epigenetic Memory by the H3K9 Methylation System 28 Modulating the epigenome of cancer cells as an innovative therapeutic approach 29 Rewiring the epigenome to change cancer chemosensitivity 30 Relación genética entre la artritis idiopática juvenil y el hipotiroidismo 31 Simulation of protein adsorption on the surface of a graphene field-effect transistor (GFET) sensor 32 Heterochromatin as the guardian of youthful epigenetic memory 33 Analysis of skin differentiation markers in severely burnt patients grafted with the UGRSKIN bioartificial skin substitute generated by tissue engineering 34 5 Construcción y visualización de un espacio cromático dental difuso mediante espacios conceptuales: PyFCS Rafa Vázquez Conejo 1, María Tejada Casado 1 2, Razvan Ghinea 1 2, Luis Javier Herrera 2 3, José Manuel Soto Hidalgo 3 1. Departamento de Óptica, Facultad de Ciencias, Universidad de Granada, Campus de Fuentenueva, s/n 18071 Granada, España 2. Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, España. 3. Departamento de Arquitectura de Computadores y Tecnología de Computadores, E.T.S.I.I.T. Universidad de Granada, s/n 18071, Granada, España. TEC09-Óptica de Biomateriales y Tejidos Rafael Vázquez Conejo | [email protected] Desarrollar un espacio cromático dental continuo y basado en la percepción humana, utilizando la teoría de los espacios conceptuales y modelos difusos, que ofrezca una representación flexible e interpretable del color, y que permita reflejar la continuidad tonal del color dental y superar las limitaciones de los métodos discretos utilizados en odontología restauradora. El sistema se desarrolló con PyFCS, una biblioteca de código abierto en Python para definir, manipular y visualizar espacios conceptuales difusos, especialmente en dominios perceptuales como el color. Permite construir categorías cromáticas mediante particionamiento tipo Voronoi, definir funciones de pertenencia y analizar imágenes. En este estudio se procesaron 16 muestras de la guía VITA Classical capturadas con cámara hiperespectral (DataColor), extrayendo valores cromáticos representativos en un espacio perceptual continuo. Se definieron funciones de pertenencia para cada tono y se construyó un conjunto difuso que modela su estructura interna. También se generaron visualizaciones 3D interactivas para explorar la geometría cromática, proximidades entre categorías y continuidad tonal. El espacio cromático difuso mostró una organización coherente con la percepción clínica del color dental, revelando transiciones graduales, solapamientos perceptuales y proximidades cromáticas relevantes. Las visualizaciones tridimensionales evidenciaron gradientes y relaciones internas no visibles en modelos discretos, obtenidas directamente de imágenes procesadas con PyFCS, facilitando un análisis detallado de la variabilidad tonal. Los espacios conceptuales difusos son una herramienta sólida y alineada con la percepción humana para representar el color dental. La integración de medidas hiperespectrales de DataColor en PyFCS permite construir espacios reproducibles, ampliables y adaptables a distintas guías clínicas, reforzando la coherencia entre datos instrumentales y percepción visual. Este enfoque sienta bases para sistemas avanzados de selección digital de color, optimización de protocolos clínicos y apoyo al diagnóstico cromático en odontología restauradora y estética. 6 DOES THE CHROMATIC ABILITY OF ONE-SHADED RESIN-BASED COMPOSITES LIMITED BY THE SURROUNDING? PRELIMINARY RESULTS Marina Chacón-Werner 1, Javier Ruiz-López 1 2, Juan C. Cardona 1 2, María M. Pérez 1 2, Razvan Ghinea 1 2, Ana Maria Andreea Ionescu 1 2 1 Department of Optics, Faculty of Science, Campus Fuentenueva, University of Granada, s/n, 18071 Granada, Spain. [email protected]; [email protected]; [email protected]; [email protected]; [email protected]; [email protected]. 2 Grupo Tec-09, Instituto de Investigación Biosanitaria ibs.GRANADA, E18011 Granada, Spain. TEC09-Óptica de Biomateriales y Tejidos Marina Chacón Werner | [email protected] One-shade composite resins are innovative biomaterials used in restorative dentistry that simplify dental shade selection due to their color-matching ability. This study evaluates the color adjustment potential of different one-shade composite resins. Cylindrical single samples (15 mm diameter and 6 mm thick) of conventional composite (Vitapan Excell ®) in three shades (A2, B2, C4) (n=27) and, of one-shade materials (Omnichroma n=3 and Omnichroma Flow n=3) were manufactured. Then, 18 samples in A2, B2 and C4 shades were restored with Omnichroma (n=9) and Omnichroma Flow (n=9), creating dual samples. The color of all single and dual samples was measured separately and under restoration conditions. Color differences CIEDE2000 (∆E00) were analysed between combinations of conventional and one-shade materials, on both type of samples. Color adaptation potential (CAP00) was used to analyzed the chromatic adaptation capability. Statistical analysis was performed using Kruskal-Wallis and Mann-Whitney test with Bonferroni correction (p<0.0001). Significant differences in CAP00 (p<0.0001) were found depending on the shade of the surrounding material, with lower performance in darker shades (C4). Significant differences were also found between one-shade materials (p<0.0001), showing Omnichroma Flow a higher color adaptation ability across all shades and giving its best results on light shades. On the other hand, conventional Omnichroma obtained better results on B2 than on A2. In conclusion, within the limitations of our study, it can be stated that Omnichroma Flow performs better CAP00 than its conventional version, especially in lighter shades. Overall, one-shade materials exhibit lower color adaptation in darker shades. Acknowledgements: This study was funded by research project C-EXP-276-UGR23, financed by Consejería de Universidad, Investigación e Innovación and by the FEDER Andalucía 2021-2027 Programme. The authors would like to thank VITA Zahnfabrik for their collaboration in the sample manufacturing process. The authors declare that they have no conflicts of interest. 7 Herramienta basada en inteligencia artificial para estimar el fracaso en la extracción testicular de espermatozoides en pacientes con azoospermia no obstructiva Sara González Muñoz 1 2, Andrea Guzmán Jiménez 1 3, Alejandra Robles Fernández 1 3, Inmaculada Higueras Serrano 1 3, F. Javier Suárez López 3, Gonzalo Borrego Yaniz 4, Miriam Cerván Martín 4, Marta Molina Romero 1 5, M. Carmen Gonzalvo 1 5, Ana Clavero Gilabert 1 5, José A. Castilla Alcalá 1 6, Nicolás Garrido Puchalt 7, Susana Seixas 8, Joao Gonçalves 9, Alexandra M. Lopes 10, Sara Larriba 11, Rogelio J. Palomino Morales 1 2, Ana Márquez 4, Lara M. Bossini Castillo 1 3, F. David Carmona López 1 3 1 Instituto de Investigación Biosanitaria ibs.GRANADA, TEC14 – Reproducción Humana y Enfermedades Hereditarias y Complejas, Granada, España. 2 Universidad de Granada, Facultad de Ciencias, Departamento de Bioquímica y Biología Molecular I, Granada, España. 3 Universidad de Granada, Facultad de Ciencias, Departamento de Genética, Granada, España. 4 Instituto de Parasitología y Biomedicina López-Neyra, Biología Celular e Inmunología, Granada, España. 5 Unidad de Reproducción UGC Obstetricia y Ginecología, Hospital Universitario Virgen de las Nieves, Granada, España. 6 Universidad de Granada, Facultad de Medicina, Departamento de Anatomía y Embriología Humana, Granada, España. 7 IVI Foundation, Health Research Institute La Fe. Servicio de Urología, Hospital Universitari I Politecnic La Fe e Instituto de Investigación Sanitaria La Fe (IIS La Fe), Valencia, España. 8 Institute of Molecular Pathology and Immunology of the University of Porto IPATIMUP, Genetic Diversity, Porto, Portugal. 9 Instituto Nacional de Saúde Dr. Ricardo Jorge, Departamento de Genética Humana, Lisboa, Portugal. 10 Istitute for Cell and Molecular Biology, Center for Predictive and Preventive Genetics, Porto, Portugal. 11 Bellvitge Biomedical Research Institute IDIBELL, Human Molecular Genetics Group, L’Hospitalet de Llobregat, España. TEC14-Reproducción Humana y Enfermedades Hereditarias y Complejas Sara González Muñoz | [email protected] La azoospermia no obstructiva (NOA), debida a un fallo grave de la espermatogénesis (SPGF), se caracteriza por la ausencia de espermatozoides en el eyaculado. Algunos pacientes pueden obtener espermatozoides mediante su extracción testicular (TESE), aunque esta técnica suele fracasar en aquellos con el síndrome de solo células de Sertoli (SCO, ausencia de línea germinal). Dado que el diagnóstico de SCO requiere de la biopsia testicular, se necesitan herramientas no invasivas que anticipen el resultado de la TESE. El objetivo fue desarrollar un modelo de inteligencia artificial basado en biomarcadores genéticos y hormonales para predecir SCO y, por tanto, el fracaso de la TESE en pacientes con NOA. Se incluyeron 293 varones infértiles con SPGF: 143 con SCO y 150 con otros fenotipos de NOA distintos a SCO. Se compararon tres modelos de machine learning (red neuronal artificial, bosque aleatorio y regresión logística) para predecir el fenotipo SCO. Los modelos incluyeron cuatro variables: (1) puntuaciones de riesgo poligénico asociadas a autoinmunidad, (2) presencia de un aminoácido específico en la proteína HLA-DRβ1, (3) niveles de hormona foliculoestimulante (FSH) y (4) ratio FSH/ hormona luteinizante (LH). El rendimiento se evaluó mediante métricas estándar en conjuntos de entrenamiento y test. La red neuronal mostró el mejor rendimiento (AUC=0,65; precisión=0,61; sensibilidad=0,62; especificidad=0,61). La presencia de serina en la molécula HLA-DRβ1 fue el predictor más relevante, reforzando la hipótesis de una base inmunológica en SCO y de la asociación de la región del HLA de clase II con este fenotipo. Las puntuaciones de riesgo poligénico de autoinmunidad fueron significativamente superiores en pacientes con SCO, y tanto la FSH como la ratio FSH/LH contribuyeron a la predicción, en concordancia con estudios previos que asocian altos niveles en estas hormonas con infertilidad. Aunque prometedor, el modelo requiere ampliación muestral y adición de nuevos biomarcadores para mejorar su robustez. Esta aproximación podría ofrecer una estimación no invasiva del resultado de la TESE y evitar procedimientos innecesarios en pacientes con alta probabilidad de fracaso. 8 Caracterización multiómica de células Sertoli adultas como base para entender la disfunción reproductiva masculina Mª del Pilar Lazúen Moreno 1, Lara María Bossini Castillo 1 2, Sara Fernández Bolívar 1, Miguel Burgos Poyatos 1, Rafael Jiménez Medina 1, Francisco Javier Barrionuevo Jiménez 1 1. Departamento de Genética e Instituto de Biotecnología, Centro de Investigación Biomédica (CIBM), Universidad de Granada 2. Instituto de Investigación Biosanitaria ibs.GRANADA TEC14-Reproducción Humana y Enfermedades Hereditarias y Complejas Mª del Pilar Lazúen Moreno | [email protected] Los trastornos del desarrollo sexual (DSD) constituyen un grupo complejode condiciones con alteraciones en la diferenciación de gónadas o genitales. La displasia campomélica (DC) es una de las formas más graves, asociada a mutaciones en SOX9, un factor de transcripción esencial para la formación esquelética y la diferenciación de células Sertoli, cuya disfunción causa fallo testicular y ambigüedad sexual. Aunque se conocen los efectos fenotípicos de la DC, aún se desconoce cómo se alteran los mecanismos que regulan la diferenciación de Sertoli. Nuestra hipótesis es que la caracterización multiómica single-cell de estas células permitirá definir estados y trayectorias cuya disrupción explica la fisiopatología de DSD como la DC. Se analizó tejido testicular en ratones control mediante tecnología single-cell multiomics (10x Genomics Multiome), que combina transcriptómica (RNA-seq) y epigenómica (ATAC-seq) en células individuales. Se secuenciaron 1,511 células, detectando 33,697 genes y 76,603 picos de accesibilidad cromatínica. El clustering bioinformático permitió identificar subpoblaciones de células Sertoli y definir etapas funcionales y de diferenciación, complementadas con análisis de genes reguladores. La trayectoria celular se reconstruyó con algoritmos de RNA velocity y Monocle3. Se validó la especificidad comparando perfiles epigenómicos con referencias de Sertoli y granulosa. Métodos estadísticos robustos permitieron explorar la correspondencia genotipo-fenotipo y posibles impactos de mutaciones DSD. Se identificaron 774 células de Sertoli en cuatro clusters reflejando un gradiente de diferenciación: inmaduras (SOX9++, TCF7L2+), intermedias (AMH+, CYP11A1+) y especializadas (KL+, EGF+). El análisis epigenómico mostró 83.1% de conservación con células de Sertoli de referencia y 68.2% de similitud con células de granulosa ovárica, sugiriendo mecanismos compartidos en determinación gonadal. Genes maestros como SOX9, GATA4 y factores meióticos se enriquecen diferencialmente entre subpoblaciones, identificando etapas vulnerables a mutaciones asociadas a DSD. Las trayectorias celulares por Monocle3 y RNA velocity presentaron 94% de concordancia, creando un atlas para proyectar perturbaciones génicas. Estos hallazgos permiten mapear correlaciones genotipo-fenotipo y definen marcadores moleculares, mostrando que la integración multiómica explica cómo mutaciones como SOX9 generan el espectro de DC y otros DSD, abriendo la vía a diagnósticos y terapias personalizadas. 9 Unveiling the immune landscape of Sertoli cell-only testes through singlecell transcriptomics in non-obstructive azoospermia Andrea Guzmán-Jiménez1,2, Sara González-Muñoz2,3, Inmaculada Higueras-Serrano1,2, Alejandra Robles-Fernández1,2, Miriam Cerván-Martín4, Marta Molina2,5, M. Carmen Gonzalvo2,5, María Luisa García-Peña6, Vicente Maldonado6, Miguel Ángel Vilches7, Andrea Espuch-Oliver8, José A. Castilla2,9, Ana Clavero2,5, Rogelio J. Palomino-Morales2,3, Cecilia Domínguez-Conde10, F. David Carmona1,2, Lara Bossini-Castillo1,2 1Departmento de Genética, Universidad de Granada, Granada, Spain. 2Instituto de Investigación Biosanitaria ibs.GRANADA, TEC14-Reproducción Humana y Enfermedades Hereditarias y Complejas,Granada, Spain. 3Departamento de Bioquímica y Biología Molecular I, Universidad de Granada, Granada, Spain. 4Institute of Parasitology and Biomedicine López-Neyra, Spanish National Research Council, Granada, Spain. 5Unidad de Reproducción, UGC Obstetricia y Ginecología, HU Virgen de las Nieves, Granada, Spain. 6Unidad de Reproducción, UGC de Obstetricia y Ginecología, HU de Jaén, Jáen, Spain. 7Ovoclinic & Ovobank, Clínicas de Reproducción Asistida y Banco de óvulos, Marbella, Málaga, Spain. 8Unidad de Reproducción Humana Asistida, Hospital Universitario Torrecárdenas, Almería, Spain. 9Departamento de Anatomía y Embriología Humana, Facultad de Medicina, Universidad de Granada, Granada, Spain. 10Human Technopole, Milan, Italy. TEC14-Reproducción Humana y Enfermedades Hereditarias y Complejas Andrea Guzmán Jiménez | [email protected] The human testis maintains an immune-privileged state that protects germ cells while hosting specialised immune populations. In non-obstructive azoospermia (NOA), particularly in the Sertoli cell-only (SCO) phenotype, this balance appears altered. Genetic associations with HLA class II molecules suggest that immune dysregulation may underlie SCO development, yet the specific immune mechanisms remain unclear. We hypothesised that SCO is driven by altered immune regulation leading to local inflammation and loss of immune privilege. To test this, we used single-cell RNA sequencing (scRNA-seq) to characterise immune and somatic landscapes in NOA testes and identify pathways linked to immune activation in SCO. Testicular biopsies from fourteen NOA patients were analysed, including five with SCO and eight with other histological subtypes. Single-cell suspensions were processed using the 10x Genomics Next GEM platform, generating more than 80,000 single-cell transcriptomes after quality control. Data were integrated and analysed in Scanpy to identify cell clusters and define germ, somatic, and immune populations. Comparative analyses between SCO and non-SCO samples in terms of immune cell enrichment, activation states, and differentially expressed genes were performed. Clustering analyses identified over twenty cellular clusters, including novel immune subpopulations. SCO testes showed marked enrichment of immune cells, notably pro-inflammatory CD8+ T cells and Type 17 helper T cells, indicating increased infiltration and activation. Gene expression patterns reflected effector and inflammatory responses, suggesting a breakdown of testicular immune privilege. These findings support an immune-mediated component in SCO pathogenesis and highlight immune dysregulation as a key factor in severe NOA, offering new perspectives for diagnosis and immunomodulatory therapies in male infertility. 10 Single-cell transcriptomic profiling uncovers seminal cellular diversity in non-obstructive azoospermia Inmaculada Higueras-Serrano 1,2, Alejandra Robles-Fernández 1,2, Sara González-Muñoz 2,3, Andrea Guzmán-Jiménez 1,2, Marta Molina 2,4, M Carmen Gonzalvo 2,4, Ana Clavero 2,4, José A Castilla 2,5, Miguel Burgos 1, Rogelio J Palomino-Morales 2,3, Lara Bossini-Castillo 1,2, F David Carmona FD1,2 1 Departamento de Genética e Instituto de Biotecnología, Centro de Investigación Biomédica (CIBM), Universidad de Granada, Granada, Spain, 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain, 3 Departamento de Bioquímica y Biología Molecular I, Universidad de Granada, Granada, Spain, 4 Unidad de Reproducción, UGC Obstetricia y Ginecología, HU Virgen de las Nieves, Granada, Spain, 5 Departamento de Anatomía y Embriología Humana, Facultad de Medicina, Universidad de Granada, Spain. TEC14-Reproducción Humana y Enfermedades Hereditarias y Complejas Inmaculada Higueras-Serrano | [email protected] Non-obstructive azoospermia (NOA), a severe form of male infertility, is defined by absent sperm in semen due to testicular failure. Based on testicular histology, NOA can present as hypospermatogenesis (HS), maturation arrest (MA), or Sertoli cell-only syndrome (SCO), each with unique testicular cellular compositions and fertility prognoses. Despite advances in assisted reproduction, predicting testicular sperm extraction (TESE) success remains limited. This study uses singlecell RNA sequencing (scRNA-seq) to profile and compare cell populations in semen from different NOA subtypes, aiming to uncover microenvironmental differences between NOA subtypes and to support development of non-invasive predictors for TESE outcomes. scRNA-seq technology was used to profile the transcriptomes of seminal cells collected from 16 Spanish men with NOA, classified as SCO (n=6), MA (n=4), or HS (n=6) based on testicular histology. Clinical TESE outcomes were collected post-biopsy to study links between TESE success and transcriptomic profiles. scRNA-seq libraries were generated using the 10x Genomics Chromium NextGEM platform, allowing high-resolution analysis of cell diversity across NOA subtypes and TESE outcomes. Following quality control, 35,646 single-cell transcriptomes were analyzed and clustered. Cell-type composition differences between TESE outcomes were assessed using permutational multivariate analysis of variance (PERMANOVA). All procedures received ethics approval and informed consent. scRNA-seq analysis of seminal cells from NOA patients revealed marked cellular heterogeneity, identifying germ cells, immune cells, and non-immunological somatic cells. The immune fraction consisted of myeloid (macrophages, monocytes, dendritic cells) and lymphoid cells (B, CD4+, CD8+ T, NK, NKT), indicating a complex local immune environment. Somatic cells included prostatic stem and epithelial cells; while germ cells were rare, in line with spermatogenic impairment. Notably, TESEsamples were enriched in macrophages and monocytes, while TESE+ samples contained more prostatic stem, epithelial, and germ cells (PERMANOVA, p=2.63×10⁻²). These cellular signatures may predict TESE outcomes and improve candidate selection for TESE, potentially sparing patients from unnecessary surgical interventions. 11 Genetic Evidence from Mendelian Randomization Supports a Causal Role of Immune-Mediated Diseases in Severe Male Infertility Alejandra Robles Fernández 1,2, Inmaculada Higueras Serrano 1,2, Andrea Guzmán Jiménez 1,2, Sara González Muñoz 2,16, José A Castilla 2,3, Ana Clavero 2,4, M Carmen Gonzalvo 2,4, Marta Molina 2,4, María Luisa García Peña 5, Vicente Maldonado 5, Miguel Ángel Vilches 6, Andrea Espuch Oliver 7, Nicolás Garrido 8, Susana Seixas 9,10, Joao Gonçalvez 11,12, Alexandra M Lopes 10,13, Elena Albani 14, Sara Larriba 15, Rogelio J Palomino Morales 2,16, F David Carmona1,2, Lara Bossini-Castillo 1,2 1 Departamento de Genética e Instituto de Biotecnología, Centro de Investigación Biomédica (CIBM), Universidad de Granada, Granada, Spain 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain 3 Departamento de Anatomía y Embriología Humana, Facultad de Medicina, Universidad de Granada, Granada, Spain 4 Unidad de Reproducción, UGC Obstetricia y Ginecología, HU Virgen de las Nieves, Granada, Spain 5 UGC de Obstetricia y Ginecología, Complejo Hospitalario de Jaén, Jaén, Spain 6 Ovoclinic & Ovobank, Clínicas de Reproducción Asistida y Banco de óvulos, Marbella, Málaga, Spain 7 Hospital Universitario Torrecárdenas, Unidad de Reproducción Humana Asistida, Almería, Spain 8 IVIRMA Global Research Alliance, IVI Foundation, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Valencia, Spain 9 i3S—Instituto de Investigaçäo e Inovaçäo em Saúde, Universidade do Porto, Porto, Portugal 10 Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal 11 Departamento de Genética Humana, Instituto Nacional de Saúde Dr Ricardo Jorge, Lisbon, Portugal 12 ToxOmics—Centro de Toxicogenómica e Saúde Humana, Nova Medical School, Lisbon, Portugal 13 CGPP-IBMC—Centro de Genética Preditiva e Preventiva, Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal 14 Division of Gynecology and Reproductive Medicine, Department of Gynecology, Fertility Center, Humanitas Research Hospital, IRCCS, Milan, Italy 15 Human Molecular Genetics Group, Bellvitge Biomedical Research Institute (IDIBELL), L’Hospitalet de Llobregat, Barcelona, Spain 16 Departamento de Bioquímica y Biología Molecular I, Universidad de Granada, Granada, Spain TEC14-Reproducción Humana y Enfermedades Hereditarias y Complejas Alejandra Robles Fernández | [email protected] Genome-wide association studies (GWAS) have implicated the major histocompatibility complex (MHC) class II region in the genetic susceptibility to Sertoli cell-only (SCO) syndrome, a severe form of non-obstructive azoospermia (NOA) characterized by a complete absence of germ cells due to profound spermatogenic failure (SPGF). Given the immunerelated genetic background of SCO, we employed a Mendelian randomization (MR) approach to explore potential causal links between autoimmune diseases and SCO, aiming to clarify the immune-mediated mechanisms underlying this extreme form of male infertility. We evaluated immune-mediated traits as potential causal factors for NOA using publicly available GWAS resuts. The NOA dataset included 1,274 men with SPGF, comprising 502 SCO cases. Causal inference was performed through twosample Mendelian randomization (2SMR) using the TwoSampleMR R package. Three complementary methods were applied: random-effects inverse variance weighted (IVW), weighted median, and MR-Egger regression. Genetic variants were selected as instrumental variables at genome-wide suggestive significance (P = 5E-05) and validated for strength (F > 10). Sensitivity analyses assessed pleiotropy via the MR-Egger intercept and heterogeneity using Cochran’s Q test. Causal effects were deemed significant at P ≤ 0.05. Using the IVW method, significant causal associations were identified between SCO and four immune-mediated traits: general autoimmune conditions (P=3.45E-02), celiac disease (P=7E-03), inflammatory bowel disease (P=4.58E-02), and Crohn’s disease (P=9E-04). The positive causal relation between them indicates that a higher genetic predisposition to immune-related disorders increases the risk of SCO, supporting an immune-mediated genetic etiology. The overlap between epithelial barrier dysfunction in celiac and Crohn’s diseases and the blood-testis barrier impairment in SCO could suggests a shared mechanism where barrier disruption exposes germ cells to immune-mediated damage. These findings support an immune-mediated genetic basis for SCO. Identifying markers of immune dysfunction may help predict unfavorable testicular sperm extraction outcomes in these patients, highlighting the clinical relevance of immune pathways for personalized counseling and management of severe male infertility. 12 Advancing CSC-Targeted Approaches in PDAC Using a Reproducible Preclinical Model of Liver Metastasis Giselle Xavier-Reis 1,2,3, Sandra Fernández Mosquera 2, Alba Hidalgo-Ramirez 1,2,3, Mar Isabel Reyes-Alcalde 4, Aitor González-Titos 1,2,3, María Tristán-Manzano 1,5,7,8, Francisco Martín 1,5,6,8, Alfonso Rúbio Navarro 1,2,5,6,9, Juan Antonio Marchal Corrales, 1,2,3,6,9,* , María Paz Zafra 1,2,3,6,9,* 1 Biosanitary Research Institute of Granada (ibs.GRANADA), University Hospitals of Granada, Av. de 30 Madrid 15, 18012 Granada, Spain. 2 Biopathology and Regenerative Medicine Institute (IBIMER), Centre for Biomedical Research (CIBM), University of Granada, 18100, Granada, Spain. 3 Department of Human Anatomy and Embryology, Faculty of Medicine, University of Granada, 18016, Granada, Spain. 4 Department of Ecology, Animal & Vegetal Biology, University of Jaen, Campus of Lagunillas, 23071, Jaen, Spain. 5 Department of Biochemistry and Molecular Biology III and Immunology, Faculty of Medicine, University of Granada, Av. de la Investigación, 11, 18006 Granada, Spain. 6 Excellence Research Unit “Modeling Nature” (MNat), University of Granada, 18016, 34 Granada, Spain. 7 LentiStem Biotech, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain. 8 Department of Genomic Medicine, Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO), PTS, Av. de la Ilustración 114, 18016 Granada, Spain. 9 BioFab i3D Lab-Biofabrication and 3D (bio)printing Singular Laboratory, University of Granada, Granada, Spain. * Corresponding authors TEC16-Terapias Avanzadas: Diferenciación, Regeneración y Cáncer Giselle Xavier-Reis | [email protected] Pancreatic Cancer(PC) is a multifactorial disease with various endocrine and non-endocrine subtypes, the most common and aggressive being pancreatic ductal adenocarcinoma(PDAC), accounting for over 92% of cases. More than 50% of PDAC patients present metastatsis in the liver, making >70% inoperable. This late diagnosis results in a 5-year overall survival rate of only 2% for metastatic PC(mPC), with a median survival of one year. Available treatments are limited to chemotherapy and palliative care. Metastasis requires tumor cells with self-renewal, plasticity, dormancy, and differentiation capabilities—key traits of cancer stem cells(CSC). Given these properties, CSC involvement in metastasis is likely. Here, we analyze CSC impact on metastatic PDAC(mPDAC) to the liver. We selected a batch of four PDAC cell lines (ASPC-1, MIAPaCa-2, BxPC-3 and PANC-1) with varying levels of aggressiveness and distinct genetic profiles to better simulate the diversity observed in human patients. For tracking purposes, all cell lines stably expressed a luciferase reporter. Following a well-established protocol, CSCs were generated from the 2D cell lines and subsequently characterized by flow cytometry. Different cell concentrations were then selected for intrasplenic injections, followed by splenectomy to properly develop liver metastasis. Initial analysis and characterization were performed using quantitative PCR, flow cytometry, and immunohistochemistry. In vitro, CSCs derived from PDAC cell lines exhibited distinct expression profiles of CD44v6, CD24 and ALDH1 confirming their heterogeneous stemness and metastatic potential. In vivo, ASPC-1–derived CSCs demonstrated the highest liver metastatic capacity compared to PANC-1, MIAPaCa-2, and BxPC-3, correlating with their aggressiveness and molecular phenotype. Detailed phenotyping of metastatic lesions revealed differential expression of CD44v6, Vim, CXCR4, and ALDH1, underscoring the relevance of CSC plasticity in driving metastatic adaptation and disease progression. Collectively, these results validate the robustness of our intrasplenic injection model to faithfully reproduce PDAC liver metastasis and provide a functional framework to investigate CSC-driven mechanisms. This model represents a valuable preclinical platform for identifying novel antimetastatic targets and developing CSC-focused precision therapies to improve outcomes in metastatic PDAC. 13 Impact of prevalent KRAS mutations in PDAC resistance to FIRINOX A. Hidalgo-Ramirez1,2, M. Reyes-Alcalde3, G. Xavier-Reis1,2, W. Sisso4,5, L. Dow4,6, A. RubioNavarro1,2,7, J. Marchal8,2,9, M. Zafra8,2,9 1Instituto de Investigación Biosanitaria ibs.GRANADA, University Hospitals of GranadaUniversity of Granada, Granada, Spain 2Centre for Biomedical Research (CIBM), University of Granada, Granada, Spain 3University of Jaen, Department of Ecology, Animal & Vegetal Biology, Jaen, Spain 4Meyer Cancer Center, Weill Cornell Medicine, New York, United States 5Weill Cornell Graduate School of Medical Science, Weill Cornell Medicine, New York, United States 6Weill Cornell Medicine, Department of Medicine, New York, United States 7University of Granada, Department of Biochemistry and Molecular Biology III and Immunology, Faculty of Medicine, Granada, Spain 8Instituto de Investigación Biosanitaria ibs.GRANADA, University of Granada, Granada, Spain 9University of Granada, Department of Human Anatomy and Embryology, Faculty of Medicine, Granada, Spain TEC16-Terapias Avanzadas: Diferenciación, Regeneración y Cáncer Alba Hidalgo Ramírez | [email protected] Pancreatic cancer is a disease with a poor prognosis due to its late diagnosis. Once diagnosed, most patients receive chemotherapy regimens as FOLFIRINOX (leucovorin, 5-fluorouracil (5-FU), irinotecan and oxaliplatin). Regrettably, resistance mechanisms are recurrent causes of treatment failure. A strategy to overcome these resistance phenotypes is to dissect them in genetically-defined model systems that take into account the heterogenous genetic landscape of these tumors.KRAS is an oncogene, which is mutated in more than 95% of patients with pancreatic ductal adenocarcinoma(PDAC). The main objective is to evaluate the mechanisms underlying acquired resistance to the combination of 3 cytostatic agents: 5-FU, SN-38 and Oxaliplatin in PDAC in the context of distinct KRAS mutations. KPOs were previously generated by transfecting LSL-KrasMUT organoids (G12D, G12V and G12R) with a plasmid expressing Cre recombinase and a sgRNA targeting Tp53. We have chosen three prevalent mutations in PDAC as KRAS-G12D (40%), KRAS-G12V (32,5%) and KRAS-G12R (18%), to then, induced chemoresistance in the different KPOs by continuously treating them with increasing concentrations of FIRINOX regimen. To assess the acquisition of chemoresistance, cell viability was measured. Then, chemoresistant (chr) and non-chemoresistant KPOs were transplanted orthotopically into the tail of the pancreas of syngeneic mice (C57Bl/6N). Harvested tumors have been profiled by immunohistochemistry and flow cytometry at final time point. We have already induced chemoresistance to FIRINOX in 3 KPO-KRASG12D, 3 KPO-KRASG12V and 3 KPO-KRASG12R lines. Preliminary data shows that in vitro chr-KPO lines tolerate a higher concentration of FIRINOX regimen. The initial characterization in vivo exhibits phenotypic changes among chr and non-chr-tumors expressing specific KRAS mutations. This preliminary data is the seed to continue our research strategy, that seeks to unravel the connection between specific-KRAS mutations, tumor heterogeneity and treatment resistance. In the near future, we hope to identify new vulnerable targets to design new therapy approaches depending on the specific KRAS mutation in the context of chemoresistance. 14 Acción de los exosomas derivados de células madre mesenquimales de médula ósea en la modulación de parámetros fibroblásticos asociados con la regeneración tisular Víctor J. Costela Ruiz 1. Facultad de Ciencias de la Salud. Departamento de Enfermería. Universidad de Granada. 2. Instituto de Investigación Biosanitaria ibs.GRANADA. TEC17-Biotejsalud Víctor Javier Costela Ruiz | [email protected] En el proceso de cicatrización los fibroblastos son esenciales, ya que sintetizan factores cruciales en la formación de la matriz extracelular. Las heridas crónicas son aquellas que presentan un proceso de inflamación prolongada, aumentando el tiempo de cicatrización y presentando un mayor riesgo de infección. Entre los abordajes novedosos en regeneración tisular, destacan terapias avanzadas como el uso de células madre mesenquimales (MSC), así como el uso de los exosomas liberados por las mismas como biomoléculas cruciales en la comunicación intercelular y la regeneración tisular. Por otro lado, compuestos fenólicos como oleocantal y oleuropeína, poseen propiedades que mejoran la regeneración tisular al estimular a los fibroblastos y promover la síntesis de colágeno. Se obtuvieron exosomas derivados de células madre mesenquimales de médula ósea sin condicionar (ExoBMMSC), y condicionados con oleocantal (OCExoBMMSC) a dosis 25M y oleuropeina (OPExoBMMSC) a dosis 100M. La caracterización incluyó técnicas como microscopía electrónica, Western blot y mediciones de Zeta Potencial. Se realizaron ensayos de tratamiento con ExoBMMSC (dosis 1,2 y 3g/mL), OCExoBMMSC (dosis 1,2 y 3g/mL) y OPExoBMMSC (dosis 1,2 y 3g/mL) sobre fibroblastos dérmicos humanos para evaluar la proliferación celular, la cicatrización in vitro mediante scratch, perfil genético de los genes COL-I, VEGF y TGF-, así como la medición de citoquinas y la proliferación un medio inflamado. Los datos se analizaron estadísticamente con ANOVA. Los resultados pusieron de manifiesto como los diferentes tratamientos mostraron una tendencia al alza en las técnicas de proliferación, migración, inhibición de la inflamación en medio inflamado in vitro, así como en la expresión de marcadores genéticos importantes en el proceso de reparación de tejidos. A destacar el efecto significativo a nivel estadístico de los tratamientos con OCExoBMMSC y OPExoBMMSC, siendo especialmente relevante en las dosis de 1 y 3g/mL respectivamente. Estos datos abren la puerta al uso de este tipo de terapia en el abordaje de lesiones tisulares de difícil cicatrización, aunque sigue siendo necesaria una mayor investigación en este sentido para determinar las vías de acción de estas nanopartículas, tanto condicionadas como sin condicionar. 21 Development of Hybrid Nanoparticles for Dual Activation Cancer Treatment via Magnetic and NIR Fields Federica Castellone1,2,3, M. Carmen Ortega-Liebana2,4,5, Arianna Bartolomei1,2,3 , Victoria CanoCortes1,2,4,5 , Stefania Nardecchia3, Alejandro R. Barroso3, Jose R. Morillas3, Juan De Vicente3, Rosario M. Sánchez-Martín1,2,4,5 1 NANOGETIC S.L., Parque Tecnológico de la salud de Granada, Avda. de la Innovación 1, 18016 Granada, Spain 2 GENYO, Centre for Genomics and Oncological Research, Pfizer/University of Granada/Andalusian Regional Government, PTS Granada, Avda. Ilustración 114, 18016 Granada, Spain 3 Department of physics, Faculty of Sciences, University of Granada, Fuente Nueva s/n 18071 Granada Spain 4 Department of Medicinal & Organic Chemistry & Excellence Research Unit of “Chemistry applied to Biomedicine and the Environment”, Faculty of Pharmacy, University of Granada, Campus de Cartuja s/n, Granada, Spain 5 Instituto de investigación biosanitaria de Granada (ibs.GRANADA), University Hospitals of Granada-University of Granada, Granada, Spain TECe18-Nanochembio Federica Castellone | [email protected] Current cancer therapies often lack the capability to selectively target tumor tissues without demaging healthy cells. Additionally, many drug delivery systems exhibit poor control over release kinetics and biodistribution. Furthermore, monotherapy approaches frequently fail to address the heterogeneity and adaptability of tumor microenvironments. Therefore, there is a critical need for integrative solutions that combine diagnostic and therapeutic functions within a single, controllable system. Our project, CoCoGel, aims to develop a nanotechnology-based platform to overcome current cancer therapy challenges. Our first approach is focused on the synthesis of hybrid nanoparticles composed of gold (Au) anchored on magnetic iron oxide (Fe₃O₄) cores, encapsulated within a silica (SiO₂) shell. Gold offers excellent biocompatibility and strong optical properties (plasmonic resonance), useful for photothermal therapies. Magnetite (Fe₃O₄) is superparamagnetic at the nanoscale, allowing magnetic guidance and heat generation under alternating magnetic fields (magnetic hyperthermia). The silica shell ensures nanoparticle stability and provides a versatile surface for functionalization with tumor-targeting ligands. A second approach utilizes gold nanorods (AuNRs) coated with a silica (SiO₂) shell and with magnetite (Fe₃O₄) nanoparticles functionalized on the external surface. Currently, we have developed several hybrid nanosystems that have been thoroughly characterized. They exhibit good biocompatibility and can be effectively activated using magnetic fields and near-infrared (NIR) radiation. By integrating multiple therapeutic modalities into a single platform, this work establishes a basis for the development of next-generation smart biomaterials for localized, effective, and minimally invasive cancer treatment. This integrative system holds promise for improved precision and efficacy in cancer treatment. 22 Nanocatalytic Platforms and Bioorthogonal Prodrug Systems to Enhance CAR-T Cell Immunotherapy in Breast Cancer Rocio Lopez-Gonzalez1,2, Maria Tristan Manzano1, Francisco Martin1, Rosario M. Sanchez-Martin1,2,3, M. Carmen Ortega-Liebana1,2,3* 1 GENYO, Pfizer/University of Granada/Andalusian Regional Government, Avda. Ilustración 114, 18016 Granada, Spain. 2 Department of Medicinal and Organic Chemistry and Unit of Excellence in Chemistry Applied to Biomedicine and Environment, Faculty of Pharmacy, Campus Cartuja s/n, University of Granada, 18071 Granada, Spain. 3 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain. TECe18-Nanochembio Rocío López González | [email protected]gr.es Chimeric antigen receptor (CAR)-T cell therapy has revolutionized cancer treatment by harnessing patients’ own immune cells to recognize and eliminate tumor-associated antigens through localized cytotoxic responses. Although highly effective against B-cell malignancies, its efficacy against solid tumors—such as breast cancer—is limited by challenges including antigen loss, an immunosuppressive tumor microenvironment and T-cell exhaustion. To overcome these hurdles, innovative strategies are urgently needed to enhance the antitumor activity of CAR-T cells directly at the tumor site. Our project integrates nanotechnology, bioorthogonal catalysis, and chemical biology to engineer precise biomedical solutions. Key objectives include designing and characterizing functionalized metal-based nanosystems for robust binding to CAR-T cells. The characterization and confirmation of this nano-decoration process were performed using flow cytometry and confocal microscopy imaging. Concurrently, a library of prodyes designed for rapid activation via nanosystem-mediated bioorthogonal catalysis was developed. We have successfully synthesized first-generation nanosystems and demonstrated their stable and selective surface binding to CAR-T cells without impairing viability. Confocal imaging and flow cytometry confirm efficient immune cell decoration, while preliminary prodye activation assays validate the catalytic activity of the platform. This work introduces a novel approach to augment CAR-T cell immunotherapy by imparting catalytic functions, enabling in situ drug activation. This strategy offers a promising means to overcome therapeutic resistance in solid tumors, advancing targeted drug delivery towards clinical translation and establishing a new paradigm for CAR-T-based smart nanomedicines. 23 Fluorescent Copper Nanocatalyst for Tracking Azide-Alkyne Cycloadditions in both solutions and cells Francisco Javier López Delgado 1 *, Mónica Rodríguez Segura 2 3 4 *, María Victoria Cano Cortés 2 3 4 *, Juan Jose Díaz Mochón 2 3 4 and Rosario María Sánchez Martín 2 3 4 1 DESTINA Genómica S.L., Avenida de la Innovación 1, Edificio BIC, Armilla 18100, Spain. 2 Department of Medicinal and Organic Chemistry, Excellence Research Unit of Chemistry Applied to Biomedicine and the Environment, School of Pharmacy, University of Granada, Campus Cartuja s/n, Granada 18071, Spain. 3 GENYO, Centre for Genomics and Oncological Research Pfizer, University of Granada and Andalusian Regional Government, PTS Granada, Avenida de la Ilustración, 114, Granada, 18016, Spain. 4 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada 18012, Spain. * These author have contributed equally to this work. TECe18-Nanochembio Francisco Javier López Delgado | [email protected] Since the introduction of click chemistry by Sharpless and Meldal and its expansion to living systems through Bertozzi’s bioorthogonal chemistry, these reactions have transformed chemical biology. Among them, the copper-catalyzed azide–alkyne cycloaddition (CuAAC) is the most prominent example, recognized with the 2022 Nobel Prize in Chemistry for its efficiency and selectivity in forming stable triazoles. Despite its potential, the biological use of CuAAC is hindered by copper toxicity, oxidation state instability, and reactive oxygen species (ROS) formation, which limit its biocompatibility. Although copper-free click reactions exist, many alkyne-containing precursors would still benefit from CuAAC chemistry if non-toxic copper systems were available. Here, we report the design and synthesis of a tracker nanocatalyst, Cu@BTTAA-Cy5-NPs, composed of copper metallofluorescent nanoparticles stabilized by the BTTAA ligand and functionalized with the fluorescent dye Cy5. These monodisperse nanoparticles display robust CuAAC catalytic activity and intrinsic fluorescence, enabling real-time monitoring of nanoparticle localization and catalytic progress within living cells. A comprehensive study revealed that Cu@BTTAA-Cy5-NPs exhibit high catalytic efficiency under mild conditions, excellent recyclability, and broad substrate scope with high yields and short reaction times. Moreover, these nanocatalysts demonstrate outstanding biocompatibility, efficient cellular uptake, and negligible toxicity, representing a new generation of non-biotoxic heterogeneous copper catalysts that combine superior catalytic performance with fluorescence tracking for intracellular and bioorthogonal CuAAC applications. 24 A Pd-modified porphyrinic MOF for combined bioorthogonal catalysis and photodynamic therapy Iria Alonso-Alves 1 2, Carolina Carrillo-Carrión 3, Belén Rubio-Ruiz 1 2 4 1 Department of Medicinal & Organic Chemistry, Faculty of Pharmacy, University of Granada, Campus Cartuja s/n, 18071, Granada, Spain. 2 GENYO, Centre for Genomics and Oncological Research, Pfizer/University of Granada/Andalusian Regional Government, PTS Granada, Avda.Ilustración 114, 18016, Granada, Spain. 3 Institute for Chemical Research (IIQ), CSIC-University of Seville, Avda. Américo Vespucio 49, 41092, Sevilla, Spain. 4 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain. TECe18-Nanochembio Iria Alonso Alves | [email protected] Standard chemotherapeutics have several limitations, including systemic side effects, poor bioavailability, and short halflives. Therefore, the development of improved anticancer strategies is mandatory. One promising approach involves the use of abiotic transition-metal catalysts (e.g. Pd) to convert inactive prodrugs into cytotoxic drugs at the tumour site. In parallel, light-triggered therapies such as photodynamic therapy (PDT) offer spatial and temporal control by generating ROS through photosensitizers, with porphyrin-based structures among the most studied. Our aim is to develop a Pd-functionalized porphyrin-based metal–organic framework (MOF) that combines the in situ depropargylation of an inactive derivative of 5-fluorouracil (Pro-5FU) and PDT under visible light irradiation. Two architectures were synthesised using PCN-222 as framework: Pd@PCN, featuring Pd(II) coordinated to the porphyrinic rings, and PCN/Pd, bearing Pd(0) nanoparticles on the MOF surface. Activation experiments were carried out under physiological conditions using Pro-Resorufin as a Pd-sensitive probe. Cellular assays were performed in BxPC3 cells, with viability measured after 5 days of treatment using PrestoBlue® reagent. Pd@PCN showed the highest catalytic activity with an 80% conversion of the off-on sensor at 24 h and demonstrated minimal toxicity even at the highest Pd concentration tested (30 μg/mL). Pd@PCN efficiently activated Pro-5FU, as confirmed by HPLC, with complete conversion to 5FU. Additionally, cells incubated with Pd@PCN and irradiated exhibited time-dependent phototoxicity. A combined chemo–PDT experiment revealed that co-treatment with Pd@PCN and Pro-5FU under irradiation produced greater inhibition of cell proliferation than individual treatments, establishing this system as a promising dual-activity nanoplatform for cancer treatment. 25 Beyond Standard Treatment: Can Exercise and Mindfulness Enhance Disability in Individuals with Chronic Primary Low Back Pain? The BACKFIT project Gavriella Tsiarleston 1 2 3, Rocío Pozuelo-Calvo 1 2 4, Pablo Molina-García, Víctor Segura-Jiménez 1,2,5,6 1 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada Spain. 2 UGC Medicina Física y Rehabilitación, Hospital Universitario Virgen de las Nieves of Granada, Granada, Spain. 3 Physical Activity for Health Promotion (PA-HELP) Research Group, Department of Physical Education and Sports, Faculty of Sport Sciences, University of Granada, Granada, Spain. 4 Department of Radiology and Physical Medicine, University of Granada, Granada, Spain. 5 GALENO research group, Department of Physical Education, Faculty of Education Sciences, University of Cádiz, Cádiz, Spain. 6 Instituto de Investigación e Innovación Biomédica de Cádiz (INiBICA), Cádiz, Spain. TECe20-Rehabilita-T: Avances e innovación en rehabilitación y promoción de la salud Gavriella Tsiarleston | [email protected] Individuals with chronic primary low back pain (CPLBP) usually present intense pain which is often accompanied by disability. The aim of this study was to compare the effects of exercise and a multimodal approach (exercise+mindfulness) with a usual rehabilitation care group on disability due to pain in individuals with CPLBP. Of 105 randomized participants, 53 individuals with CPLBP aged 52±9 years attended >65% of the program and were analyzed: control group (CG, n=19), exercise group (EG, n=14) and exercise+mindfulness group (EMG, n=20). All groups completed an 8-week intervention twice weekly (45 minutes/session). The CG received usual care (stretching and motor control exercises). The EG and EMG performed supervised muscle-strengthening and motor control exercise. Additionally, the EMG attended weekly mindfulness session (2.5 hours/session). Disability due to pain was assessed with the Oswestry Disability Index. Assessments were performed preand postintervention and after a 2month detraining period (re). Repeated measures (Wilcoxon test) showed that EG (median (Me)=-12.00; Interquartile range (Q1-Q3)= -16.44, -5.00; p=0.003) and EMG (Me= -3.11; Q1-Q3= -12.00, 0.00; p=0.005) reduced disability due to pain after the intervention. CG did not show improvements (Me= -8.00; Q1-Q3= -11.11, -2.00; p>0.05). After the detraining period, no significant intra-group differences were found compared to the baseline (all, p>0.05). Kruskal-Wallis test showed no significant between-group differences in post-intervention and follow-up changes from baseline for any of the outcomes (all, p>0.05). To conclude, both exercise alone and exercise and mindfulness approach were effective compared to usual rehabilitation care in reducing disability due to pain in individuals with CPLBP. Future research is warranted to explore whether multimodal interventions incorporating higher exercise intensity, yield superior effects in this population. 26 Melatonin Ameliorates Heart Failure by Improving Ventricular Remodeling and Ejection Fraction in Male ZDF Rats José Juan Ángel Morillo 1, Diego Salagre Simón 1, Silvia López Fernández 2, Ahmad Agil Abdalla 1 1 Department of Pharmacology, BioHealth Institute Granada (ibs.GRANADA), Neuroscience Institute (CIBM), School of Medicine, University of Granada, Granada, Spain. 2 Cardiology Department, Heart Failure Unit, Virgen de las Nieves University Hospital, BioHealth Institute Granada (ibs.GRANADA), Granada, Spain. MP-19 Nutrición, Metabolismo, Crecimiento y Desarrollo José Juan Ángel Morillo | jj[email protected]aen.es Heart failure (HF) is a clinical syndrome in which the heart is unable to pump sufficient blood to meet the metabolic demands of the body. The prevalence of HF is higher in men, and numerous studies have reported marked sex-related differences in both cardiac structure and function. Melatonin is a hormone that regulates circadian rhythms and exhibits pleiotropic cardioprotective properties. This molecule mitigates cardiac injury and physiological disturbances through its potent antioxidant, anti-inflammatory, and anti-apoptotic effects. In addition, several studies using muscle samples have demonstrated that melatonin modulates mitochondrial dynamics, thereby contributing to the restoration of mitochondrial function and energy production. In this study, 9-week-old male obese diabetic Zücker rats (ZDF) and their lean littermates (ZL) were divided into four subgroups (n=4): control (C), melatonin-treated (M), dapagliflozin-treated (D), and dapagliflozin and melatonin-treated (DM). Treatments were administered orally at a dose of 10 mg/kg/day for 17 weeks. Cardiac function was assessed by echocardiography and plethysmography. Western blot analysis were performed on the left ventricle (LV) to study mitochondrial dynamics and calcium metabolism. Histological stains (H&E, Masson's trichrome, picrosirius red, and oil red O) were used to analyze remodeling, fibrosis, inflammation, hypertrophy, and lipid accumulation. Echocardiography revealed increased ejection fraction and ventricular volumes in M, D, and DM ZDF rats, while plethysmography showed reduced systolic and diastolic pressures across all treatments. Histological analyses indicated that melatonin and dapagliflozin decreased myocardial fibrosis, lipid accumulation, and cardiomyocyte hypertrophy, obtaining synergistic effects in DM ZDF rats. Cardiac metabolism and mitochondrial function were also improved in all groups, with enhanced outcomes in DM ZDF rats. Treatments also reduced mitochondrial fission and promoted fusion, with M ZDF rats yielding greater effects in fusion promotion. The combined treatment further reduced mitochondrial fission and enhanced fusion. In conclusion, melatonin plays an important cardioprotective role by reducing ventricular remodeling and mitochondrial dysfunction, and its combination with dapagliflozin may represent a novel and more effective therapeutic strategy for the management of heart failure. 27 Conexiones genéticas entre la depresión mayor y la obesidad: un enfoque desde la secuenciación de nueva generación José Luis Ruiz 1,2,3, Esther Molina 4,2,3, Jorge Antonio Cervilla 5,2,3, Blanca Gutiérrez 5,2,3, Esperanza de Santiago 6, Maria Luisa Cañadas 7, Santiago Posik 8,5, Coral del Val 9,3, Margarita Rivera 1,2,3 1 Departamento de Bioquímica y Biología Molecular II - Facultad de Farmacia - Universidad de Granada 2 Instituto de Neurociencias Federico Olóriz - Centro de Investigación Biomédica (CIBM) - Parque Tecnológico de la Salud (PTS) - Granada 3 Instituto de Investigación Biosanitaria ibs.GRANADA 4 Departamento de Enfermería, Facultad de Ciencias de la Salud - Universidad de Granada, 5 Departamento de Psiquiatría, Facultad de Medicina - Universidad de Granada 6 Centro Pfizer - Universidad de Granada - Junta de Andalucía de Genómica e Investigación Oncológica (GENyO) - Unidad de Genómica. 7 Queen's University Belfast - The University of Bristol, University of Bristol 8 Hospital Clínico Universitario SAN Cecilio - Granada. 9 Departamento de Ciencias De La Computación e Inteligencia Artificial - Instituto Andaluz Interuniversitario En Ciencia De Datos e Inteligencia Computacional (DaSCI) – Granada E05-Psiquiatría Bioambiental José Luis Ruiz Moreno | [email protected] La depresión mayor (DM) y la obesidad son trastornos complejos, altamente prevalentes y con un impacto significativo en la salud pública. Diversas evidencias epidemiológicas y genéticas respaldan una relación bidireccional entre ambas condiciones, probablemente mediada por mecanismos compartidos de disfunción neuroendocrina, inflamación crónica y vulnerabilidad genética. Sin embargo, los correlatos genéticos específicos de este fenotipo comórbido permanecen poco caracterizados. La secuenciación de exoma completo (WES) constituye un enfoque de nueva generación ideal para identificar variantes genéticas comunes, raras o novedosas asociadas a la DM, la obesidad y su fenotipo comórbido. En este proyecto se realizará un análisis WES en una submuestra de 654 individuos del estudio PISMA-ep. la submuestra PISMA-ep de 654 individuos se divide en cuatro grupos: i) individuos con DM sin obesidad; ii) con obesidad sin DM; iii) con fenotipo comórbido; iv) controles sanos. Los datos de secuenciación se procesarán con un pipeline estandarizado que incluirá control de calidad, alineamiento y refinamiento, anotación funcional y filtrado de variantes raras y posteriormente se optimizará. Se realizarán análisis de rutas biológicas para ver los procesos implicados en las variantes identificadas y análisis funcionales a nivel de variante y gen. Posteriormente se validarán los resultados con bases de datos públicas y validación experimental. Se espera que la identificación y caracterización de perfiles genéticos de susceptibilidad contribuya a reconocer a la población en riesgo y desarrollar estrategias de prevención dirigidas y personalizadas, lo que no solo mejorará la calidad de vida de los pacientes y reducirá comorbilidades, sino que también optimizará la calidad asistencial y generará un impacto positivo en la eficiencia del Sistema de Salud. Se prevé identificar rutas metabólicas y de señalización no descritas previamente en relación con la enfermedad. Este hallazgo podría ampliar el conocimiento sobre los mecanismos fisiopatológicos subyacentes y revelar nuevos puntos de intervención terapéutica, abriendo la posibilidad de reposicionar fármacos ya existentes o diseñar terapias dirigidas. Asimismo, se plantea el desarrollo de un pipeline computacional reproducible que integre las principales fases del análisis de datos ómicos, facilitando su reutilización por otros grupos. 28 Dissecting the Molecular Basis of Epigenetic Memory by the H3K9 Methylation System Jiajun Xie 1 2 3, Joan Reinés Domingo 1 3, David Landeira Frías 1 2 3 1 Centro de Genómica e Investigación Oncológica (GENyO) 2 Facultad de Farmacia, Universidad de Granada 3 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain Ae21-Epigenética en Envejecimiento y Cáncer Jiajun Xie | [email protected] Loss of cell identity caused by epigenetic instability is emerging as a fundamental hallmark of ageing and disease. As therapeutic manipulation of the epigenome becomes a frontier in modern medicine, understanding how cells preserve transcriptional memory is essential, yet the molecular basis of this process remains elusive. Methylation of histone H3 at lysine 9 constitutes a key repressive chromatin mark associated with heterochromatin formation and transcriptional silencing of genes and repetitive elements. The faithful transmission of these repressive states through cell division requires epigenetic memory mechanisms that ensure the inheritance and maintenance of specific expression and silencing programs across cell generations. In this work, we examined multiple components of the H3K9 methylation machinery using diverse molecular and cellular approaches to dissect their contributions to the maintenance and inheritance of epigenetic repression. Our findings provide new insights into how H3K9-dependent mechanisms safeguard transcriptional states during cell division, potentially informing future strategies to counteract epigenetic dysfunctions underlying ageing and disease. 29 Modulating the epigenome of cancer cells as an innovative therapeutic approach Andrea Serrano-Prados 1 2 3 *, Adrián Castillo-Linde 1 2 3, Rita Caracuel-Peramos 1, Iván FernándezRengel 1 2 3, Efres Belmonte-Reche 1 2 3, María Alcázar-Fabra 1 2 3, Amador Gallardo 1 2 3, María Del Carmen Plaza-Calonge 1, Juan Carlos Rodríguez-Manzaneque 1, Lourdes López-Onieva 1 3 4 and David Landeira 1 2 3 1 Centre for Genomics and Oncological Research (GENYO), Avenue de la Ilustración 114, 18016 Granada, Spain 2 Department of Biochemistry and Molecular Biology II, Faculty of Pharmacy, University of Granada, Granada, Spain 3 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain 4 Department of Biochemistry and Molecular Biology I, Faculty of Science, University of Granada, Granada, Spain Ae21-Epigenética en Envejecimiento y Cáncer Andrea Serrano Prados | [email protected] Current cancer treatments primarily aim to remove malignant cells or induce their cytotoxicity through surgery, radio-, chemo-, or immunotherapy. However, these interventions often cause substantial toxicity in healthy tissues and incomplete therapeutic responses, leading to patient distress and economic burden on public health system. Epigenetic factors have emerged as key druggable targets in cancer and small-molecule inhibitors of epigenetic regulators (epidrugs) are being developed or approved for clinical use. Nonetheless, their clinical success has so far been limited, likely in part due to ineffective therapeutic design. Hence, innovative approaches are required to fully harness the potential of epigenetic therapy in cancer. Here, we provide new insights toward achieving this goal. By combining transcriptomics, epigenomics, cancer cell lines, and mouse xenograft models of non-small cell lung cancer, melanoma, multiple myeloma, and acute myeloid leukaemia, we aim to analyse the downstream effect of treating cancer cells with small-molecule clinically approved epidrugs on their epigenome, transcriptome, and tumorigenic capacity. We exhibit the results of new posible experimental designs with epidrugs to treat cancer cells and fully take advantage of the potential of those small-molecule inhibitors, which could even be combined with current therapies. 30 Rewiring the epigenome to change cancer chemosensitivity Adrián Castillo-Linde 1 2 3, Andrea Serrano-Prados 1 2 3, Rita Caracuel-Peramos 1, Iván FernándezRengel 1 2 3, Efres Belmonte-Reche 1 2 3, María Alcázar Fabra 1 2 3, Amador Gallardo 1 2 3, María Del Carmen Plaza-Calonge 1, Juan Carlos Rodríguez-Manzaneque 1, Lourdes López-Onieva 1 3 4 and David Landeira 1 2 3 1 Centre for Genomics and Oncological Research (GENYO), Avenue de la Ilustración 114, 18016 Granada, Spain 2 Department of Biochemistry and Molecular Biology II, Faculty of Pharmacy, University of Granada, Granada, Spain 3 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain 4 Department of Biochemistry and Molecular Biology I, Faculty of Science, University of Granada, Granada, Spain Ae21-Epigenética en Envejecimiento y Cáncer Adrián Castillo Linde | [email protected] Cancers represent the second leading cause of mortality occurring worldwide, accounting for up to 10 million deaths in 2020. This situation has driven the development of numerous therapeutic strategies that, while initially effective, may lose efficacy over time due to the emergence of resistance, progressively narrowing the range of drugs available for the treatment of a given tumor. Emerging evidence reveals epigenetics plays a significant role in cancer behavior, controlling processes such as chemoresistance or even acting as a major driver of disease. Therefore, and although epigenetic therapy was first designed seeking cytotoxic activity, we believe it has the potential, through changes in the epigenome, to affect how cancer respond to other cytotoxic drugs and synergize with them. Using epigenomic analysis and mouse xenograft assays we aim to study the impact of the epigenome on drug response. Our results suggest that treatment with epigenetic drugs may induce changes in how cancer cells respond to different chemotherapeutic agents. 37 Prevascularized fibrin-agarose skin substitutes with mesenchymal stem cells for enhanced biointegration of skin models Chato-Astrain J1,2, Ávila-Fernández P1,2, Sánchez-Porras D1,2, García-García OD1,2, Ortiz-Arrabal O1,2, Bermejo-Casares F1, Crespo PV1,2, Alaminos M1,2, Garzón I1,2, Martín-Piedra MA1,2 1 Tissue Engineering Group, Department of Histology, University of Granada, Spain 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain TEC-03 Ingeniería Tisular Jesús Chato Astrain | [email protected] Background: Rapid and stable vascularization is critical for the success of bioartificial skin substitutes in severely burned patients, as it ensures nutrient supply, oxygenation, and integration with host tissue (1). UGRSKIN, based on fibrin-agarose hydrogels, has demonstrated clinical efficacy in restoring the epithelial barrier (2), but its biointegration could be further improved by incorporating prevascularized cellular components. Mesenchymal stem cells (MSC) are known to promote angiogenesis, modulate inflammation, and support extracellular matrix remodeling. Combining MSC with endothelial cells within the scaffold under pro-angiogenic conditions may accelerate neovascularization and enhance graft survival. Methods: Fibrin-agarose scaffolds were fabricated and combined with endothelial cells and MSC (bone marrow and adipose-derived), either undifferentiated or pre-differentiated toward vascular phenotypes. Constructs were cultured using pro-angiogenic conditions and induction media before implantation. Biointegration was assessed by histology and immunohistochemistry at 7 days post-grafting in experimental models. Results and Conclusions: Prevascularized fibrin-agarose skin substitutes showed accelerated neovascularization and improved integration compared to standard UGRSKIN. MSC promoted the formation of branched vascular networks and enhanced extracellular matrix deposition. These findings suggest that incorporating MSC into fibrin-agarose scaffolds represents a promising strategy to develop next-generation skin substitutes with superior regenerative potential for burn patients. Acknowledgements: Supported by grants C-CTS-032-UGR23 and DGP_PIDI_2024_00361, programa operativo FEDER Andalucía 2021–2027 and Consejería de Universidad, Investigación e Innovación, Junta de Andalucía and Plan Propio de Investigación y Transferencia de la Universidad de Granada 2023. Supported by Instituto de Salud Carlos III, Ministry of Science, Innovation and Universities, Grant FIS PI25/00002, and co-funded by the European Union. References: 1. Rademakers T, Horvath JM, van Blitterswijk CA, LaPointe VLS. Oxygen and nutrient delivery in tissue engineering: Approaches to graft vascularization. J Tissue Eng Regen Med. 2019 Oct;13(10):1815-1829 2. Martin-Piedra MA, Carmona G, Campos F, Carriel V, Fernández-González A, Campos A, Cuende N, Garzón I, Gacto P, Alaminos M. Histological assessment of nanostructured fibrin-agarose skin substitutes grafted in burnt patients. A timecourse study. Bioeng Transl Med. 2023 7;8(6):e10572 38 Signaling pathways orchestrating ex vivo maturation of human bioengineered dermo-epidermal skin substitutes Ávila-Fernández P1,2, Etayo-Escanilla M1,2, Campos F1,2, Sánchez-Porras D1,2, García-García OD1,2, Martín-Piedra MA1,2, Garzón I1,2, Campos A1,2, Alaminos M1,2, Chato-Astrain J1,2 1 Tissue Engineering Group, Department of Histology, University of Granada, Spain 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain TEC-03 Ingeniería Tisular Paula Ávila Fernández | [email protected] Background: Previous reports demonstrated that the fibrin-agarose bioartificial skin substitute UGRSKIN may be clinically useful for the treatment of severely burnt patients (1). However, the molecular mechanisms governing ex vivo maturation of this skin substitute remain poorly understood. Determining signaling dynamics during culture is essential to guide strategies enhancing biomimicry and predicting graft performance. Methods: Two human skin substitutes were analyzed: an epithelial substitute (ESS) and a stromal-epithelial substitute containing fibroblasts (SESS). Constructs were cultured under air-liquid conditions for 14 days. Protein arrays were used for profiling key functional (proliferation, epidermal/dermal differentiation, stress/apoptosis, inflammation) and signaling pathways (MAPK, AKT, JAK/STAT, NF-κB, TGF-β). Results and Discussion: SESS promoted a more balanced and physiological maturation compared to ESS models. Stromalepithelial constructs activated MAPK and AKT pathways (ERK1/2, RAF-1, PDK1, GSK3β), supporting proliferation and tissue organization, while late TGF-β signaling (SMAD1, c-Jun) enhanced dermal remodeling and epidermal differentiation. Furthermore, SESS attenuated excessive inflammatory and stress responses linked to JAK/STAT (STAT3) and NF-κB, creating a signaling environment favorable for structural stability and functional maturation. This coordinated signaling environment in stromal-epithelial models suggests that fibroblast-derived cues are essential for guiding epidermal differentiation and structural stability. Conclusions: Mapping signaling pathways during ex vivo development provides actionable criteria for optimizing UGRSKIN manufacturing which could improve predictability and clinical outcomes of human bioengineered skin for use in burnt patients. Acknowledgements: Supported by grants C-CTS-032-UGR23 and DGP_PIDI_2024_00361, programa operativo FEDER Andalucía 2021–2027 and Consejería de Universidad, Investigación e Innovación, Junta de Andalucía and Plan Propio de Investigación y Transferencia de la Universidad de Granada 2023. Supported by Instituto de Salud Carlos III, Ministry of Science, Innovation and Universities, Grant FIS PI25/00002, and co-funded by the European Union. References 1. Martin-Piedra MA, et al. Histological assessment of nanostructured fibrin-agarose skin substitutes grafted in burnt patients. A time-course study. Bioeng Transl Med. 2023;8(6):e10572 39 Ex vivo extracellular matrix maturation of UGRSKIN fibrin-agarose skin substitutes resembles the development process of the native skin Ávila-Fernández P1,2, Campos F1,2, García-García OD1,2, Ortiz-Arrabal O1,2, Sánchez-Porras D1,2, Etayo-Escanilla M1,2, Bermejo-Casares F1, España-López A3, Alaminos M1,2, Chato-Astrain J1,2 1 Tissue Engineering Group, Department of Histology, University of Granada, Spain 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain 3 Department of Stomatology, University of Granada, Spain TEC-03 Ingeniería Tisular Paula Ávila Fernández | [email protected] Background: The extracellular matrix (ECM) plays a fundamental role in supporting skin structure and function, influencing cell adhesion, signaling, and tissue regeneration (1). Several models of human bioartificial skin substitutes have been developed, including UGRSKIN, based on fibrin-agarose hydrogels, which proved to be safe and clinically useful for the treatment of burnt patients. However, understanding the complex process of ECM maturation that takes place during ex vivo development of this artificial tissue is essential to optimize graft quality and integration. In the present work, we carried out an ex vivo analysis of the ECM at different time points to characterize the development process of UGRSKIN. Methods: UGRSKIN substitutes were generated using a cellular stromal layer based on fibrin-agarose biomaterials with fibroblasts within and a stratified epithelium on top. This artificial tissue was cultured under controlled conditions and sequentially analyzed at different time points. ECM maturation was analyzed by immunohistochemistry and a molecular profiling of key components, including collagens (I, IV, VII), proteoglycans (decorin, agrin, perlecan), and basement membrane proteins (fibronectin, nidogen) was carried out. Activation of signaling pathways such as TGF-β was also assessed. Results and Conclusions: UGRSKIN dermo-epidermal skin models exhibited a progressive and organized deposition of ECM molecules resembling the native dermis, with early basement membrane formation and increased expression of structural proteins over time. These findings confirm that epithelial-stromal interaction is critical for ECM development and suggest that ex vivo maturation of UGRSKIN enhances its molecular similarity to normal skin, potentially improving biointegration and clinical performance in burn care. These results contribute to understanding the positive effects of UGRSKIN applied to severely burnt patients. Acknowledgements: Supported by grants C-CTS-032-UGR23 and DGP_PIDI_2024_00361, programa operativo FEDER Andalucía 2021–2027 and Consejería de Universidad, Investigación e Innovación, Junta de Andalucía and Plan Propio de Investigación y Transferencia de la Universidad de Granada 2023. Supported by Instituto de Salud Carlos III, Ministry of Science, Innovation and Universities, Grant FIS PI25/00002, and co-funded by the European Union. References: 1. Park E, Kim HG, Un Y, Oh ES. Spatial and Functional Roles of Syndecans in Skin Wound Healing. Int J Mol Sci. 2025 Oct 30;26(21):10571 40 Optimizing clinical-grade MSC expansion: differential efficacy of platelet lysate and human plasma in adipose and bone marrow stem cells Pérez-Ortiz G1, Rejón-Camacho A1, Ávila-Fernández P1,2, Etayo-Escanilla M1,2, Aguilar-Pérez S1, González-Gallardo C1,2,3, Campos F1,2, Chato-Astrain J1,2, Alaminos M1,2, Martín-Piedra MA1,2, Garzón I 1,2 1 Tissue Engineering Group, Department of Histology, University of Granada, Spain 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain 3 Division of Ophthalmology, University Hospital Clínico San Cecilio, Granada, Spain TEC-03 Ingeniería Tisular Ingrid Garzón | [email protected] Background: Human mesenchymal stem cells (MSC) are increasingly used in tissue engineering protocols, due to their differentiation potential and proliferation capacity (1). For use in advanced therapies, bioartificial tissues must be fabricated using good manufacturing practices (GMP) guidelines, which strongly recommend the use of non-xenogeneic materials and reagents. However, current cell culture and tissue engineering protocols are largely dependent on the use of fetal bovine serum (FBS) supplementation. The use of this bovine product harbors several drawbacks, such as batch-to-batch variability, risk of disease transmission and potential immune reactions. One of the possible alternatives to the use of FBS is supplementation of the culture media with human-derived components, such as the human plasma (HP) and platelet lysates (PL). In this study, we compared the efficacy of HP and PL applied to the culture and expansion of two types of human MSC commonly used in tissue engineering for the generation of heterotypical models of the human oral mucosa, palate, cornea and skin by tissue engineering. Methods: First, two types of human MSC were isolated from human tissue biopsies using enzymatic methods, as previously reported (2). Then, cells were cultured for several cell passages using DMEM medium supplemented with different concentrations of HP or PL, using the same medium with FBS as a control gold-standard group. Results were analyzed to determine the effects of each culture condition on cell survival, as determined by LIVE/DEAD analysis and cell proliferation. Results: In general, all culture conditions allowed the efficient growth and proliferation of the MSC cultures, with more than 90% cell viability in all cases, suggesting that the use of HP and PL might support cell viability in a comparable manner to FBS. Cell proliferation varied among groups, with PL showing the highest rates of cell proliferation, supporting cell passaging without any detectable alterations. Conclusions: The use of LP and PH is a safe promising alternative to FBS for the clinical-grade expansion of human MSC, fulfilling the requirements for tissue engineering translational products. Future studies should determine the real potential of these xenogeneic-free methods to generate bioartificial substitutes of the human oral mucosa, palate, cornea and skin generated with alternative MSC cell sources. Acknowledgeents: Supported by Instituto de Salud Carlos III (ISCIII), Ministry of Science, Innovation and Universities grants FIS PI23/00335, FIS 24/00006, FIS PI25/00002, ICI19/00024-BIOCLEFT and ICI21/00010-NANOULCOR, cofunded by the European Union. Supported by DGP_PIDI_2024_00361, grant DGP_PIDI_2024_01347 and C-CTS-032UGR23, programa operativo FEDER Andalucía 2021–2027, Consejería de Universidad, Investigación e Innovación, Junta de Andalucía and University of Granada, Spain. References: 1. Garzon I, Chato-Astrain J, Campos F, Fernandez-Valades R, Sanchez-Montesinos I, Campos A, Alaminos M, D'Souza RN, Martin-Piedra MA. Expanded Differentiation Capability of Human Wharton's Jelly Stem Cells Toward Pluripotency: A Systematic Review. Tissue Eng Part B Rev. 2020 Aug;26(4):301-312 2. Vico M, Rodríguez-Morata A, Garzón I, Campos F, Jaimes-Parra B, Pérez-Köhler B, Buján J, Alaminos M, SánchezQuevedo MC. Cell viability evaluation of transdifferentiated endothelial-like cells by quantitative electron-probe X-ray microanalysis for tissue engineering. Histol Histopathol. 2015 Nov;30(11):1333-40 41 Development of bioartificial tissues functionalized with fresh human bone particles for oral mucosa and palate tissue engineering applications Martín-Piedra MA1,2, España-López A3, Etayo-Escanilla M1,2, Pérez-Ortiz G1, Ávila-Fernández P1,2, Sánchez-Porras D1,2, Ortiz-Arrabal O1,2, García-García OD1,2, Fernández-Valadés R1,2,4, Campos F1,2, Garzón I1,2 1 Tissue Engineering Group, Department of Histology, University of Granada, Spain 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain 3 Department of Stomatology, University of Granada, Spain 4 Division of Pediatric Surgery, University Hospital Virgen de las Nieves, Granada, Spain TEC-03 Ingeniería Tisular Ingrid Garzón | [email protected] Introduction: Tissue engineering is focused on the design and development of biomimetic substitutes capable of promoting tissue regeneration in complex environments such as the oral cavity and, specifically, in those situations where a bone defect is present (1), such as cleft palate (2). This research aimed to develop and characterize an artificial oral mucosa substitute based on fibrin-agarose hydrogels functionalized with living human bone particles able to partially reproduce the structure of the oral mucosa in contact with palatal bone, such as the human hard palate, and to evaluate these functionalized oral mucosa substitutes at different levels. Methods: A substitute of the human oral mucosa lamina propria was generated using fibrin-agarose hydrogels with oral fibroblasts immersed within. These substitutes were functionalized with small fragments of the human maxillary bone obtained from patients subjected to dental implants. These artificial tissues were kept in culture under controlled conditions for three weeks and evaluated at different times. Histological analyses were performed to assess morphological structure using hematoxylin-eosin and the presence of of key extracellular matrix (ECM) molecules was assessed by histochemistry. Results and conclusions: Functionalized human oral mucosa substitutes showed a progressive colonization of cells that migrated from the bone particles to the biomaterial after two weeks of culture, suggesting that the cells in bone particles remains viable and were able to proliferate and migrate ex vivo. These artificial tissues also showed some non-fibrillar components of the ECM at the extracellular compartment, suggesting that the cells in the artificial tissue were able to synthesize and release key ECM components, although the artificial lamina propria was still immature and not fully biomimetic to the native tissue. These findings support the potential of fibrin-agarose oral mucosa substitutes functionalized with bone particles as a promising strategy for short-term development of constructs with improved potential to integrate at bone surfaces, such as the hard palate, although extended maturation times may be required to achieve full matrix organization suitable for clinical application in oral regenerative therapies. Acknowledgements: Supported by Instituto de Salud Carlos III, Ministry of Science, Innovation and Universities, Grant FIS PI24/00006, and co-funded by the European Union. References: 1. Zhao G, Wang G, Paka Lubamba G, Bao M, Li C, Gao N. Review of Hard Palate Lipoma and Retrospective Clinicopathological Analysis of Patients With Oral Lipomas. J Craniofac Surg. 2025 May 1;36(3):943-946 2. Martín-Del-Campo M, Rosales-Ibañez R, Rojo L. Biomaterials for Cleft Lip and Palate Regeneration. Int J Mol Sci. 2019 May 2;20(9):2176 42 Development of cryopreservation protocols for long-term storage of human artificial tissues generated by tissue engineering Aguilar-Pérez S1, Etayo-Escanilla M1,2, González-Gallardo C1,2,3, Rejón-Camacho A1, Ávila-Fernández P1,2, Ortiz-Arrabal O1,2, Campos F1,2, Alaminos M1,2, Chato-Astrain J1,2, Garzón I1,2, Martín-Piedra MA1,2 1 Tissue Engineering Group, Department of Histology, University of Granada, Spain 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain 3 Division of Ophthalmology, University Hospital Clínico San Cecilio, Granada, Spain TEC-03 Ingeniería Tisular Miguel Ángel Martín Piedra | [email protected] Introduction: Severe conditions affecting human tissues with limited regeneration potential are very difficult to manage, and the use of tissue grafts is often necessary. Tissue engineering offers an alternative strategy based on the generation of tissues consisting of an artificial stroma and an overlying epithelium. However, the half-life of these artificial tissues is very short. In this scenario, cryopreservation becomes an appealing option (1), as it may enable large-scale production, long-term storage and rapid clinical deployment of engineered constructs in patients with severe tissue defects. The present study analyses the efficiency of different cryopreservation protocols and how these protocols may influence the metabolic activity of artificial tissues, emphasising on their proliferative potential and their ability of matrix remodelling. Methods: Bilayered artificial tissues containing stroma and epithelial layers were engineered using fibrin-agarose scaffolds. These artificial tissues were then cryopreserved in different cryoprotective agents containing DMSO, glycerol, trehalose or other cryopreservation compounds, and stored at three temperature conditions (4ºC; -20ºC; -80ºC) for increasing periods of time. Two control groups were included: non-cryopreserved fresh artificial tissues (CTR+) and artificial tissues cryopreserved in PBS (CTR-). After thawing, immunohistochemical assays were performed to quantify total cell content and the percentage of cells showing positive expression for proteins related to cell proliferation and tissue remodelling. Results and conclusions: In general, all cryopreservation agents were partially successful in terms of tissue preservation and cell proliferation, although none of them were fully functional. We observed trehalose showed appropriate results at 4º C. Nevertheless, at lower temperatures (-20 and -80ºC), other reagents were more efficient and kept the proliferative status of the cells. Regarding matrix remodelling, we found a decrease at the longest periods of time. These findings suggest that cryopreservation could be a valuable option, and optimization of the protocols showing the most favorable preliminary results could contribute to develop more efficient cryopreservation methods for long-term storage of human bioartificial tissues. Acknowledgements: Supported by Instituto de Salud Carlos III (ISCIII), Ministry of Science, Innovation and Universities, grants FIS PI23/00335, FIS 24/00006, FIS PI25/00002, ICI19/00024-BIOCLEFT and ICI21/00010-NANOULCOR, cofunded by the European Union. Supported by DGP_PIDI_2024_00361, grant DGP_PIDI_2024_01347 and C-CTS-032UGR23, programa operativo FEDER Andalucía 2021–2027, Consejería de Universidad, Investigación e Innovación, Junta de Andalucía and University of Granada, Spain. References: 1. Budharaju H, Sundaramurthi D, Sethuraman S. Biofabrication & cryopreservation of tissue engineered constructs for on-demand applications. Biofabrication. 2024 Sep 23;16(4) 43 Characterization of novel natural biomaterials for corneal tissue engineering Alaminos M1,2, Pereira-Martínez J1, Muñoz-Hurtado J1, Tejeda-Casado M2,3, Ionescu A2,3, Cardona JC2,3, Rodríguez-Escalera EA1, Pérez-Gómez MM2,3, Garzón I1,2 1 Tissue Engineering Group, Department of Histology, University of Granada, Spain 2 Instituto de Investigación Biosanitaria ibs.GRANADA, Granada, Spain 3 Department of Optics, University of Granada, Spain TEC-03 Ingeniería Tisular Miguel Ángel Martín Piedra | [email protected] Background: Different types of biomaterials have been used to reproduce the structure of the human cornea by tissue engineering. Among others, natural biomaterials typically show adequate biocompatibility both ex vivo and in vivo (1) and are excellent candidates for use in corneal tissue engineering. In the present work, we have evaluated a transparent material obtained from marine invertebrates used as a scaffold biomaterial for cornea tissue engineering. Methods: First, we obtained a natural biomaterial from the internal organs of marine invertebrates. This biomaterial was washed several times in PBS and evaluated histologically. Then, corneal epithelial cells were cultured on top of the biomaterial to determine the ex vivo biocompatibility of this biomaterial, and immunohistochemical analyses were carried out after 4 days of culture to determine cell phenotype. Results and conclusions: We first found that the natural biomaterials were very transparent, and let most of the incoming visible light go through the biomaterial without alterations. Then, histological analyses found that the biomaterial was very homogeneous, and consisted of multiple parallel layers of material containing limited amounts of collagen. When cells were cultured on these biomaterials, we found a good integration, with cells forming an epithelial cell layer on top with positive expression of corneal crystallins, with cells showing high viability. In general, these results suggest that these marine-derived natural biomaterials may have putative usefulness for use in corneal tissue engineering. Acknowledgements: This work was supported by Instituto de Salud Carlos III, Spanish Ministry of Science, Innovation and Universities, Grants FIS PI23/00335 and ICI21/00010 (NANOULCOR), and co-funded by the European Union. References: 1. Majid QA, Fricker ATR, Gregory DA, Davidenko N, Hernandez Cruz O, Jabbour RJ, Owen TJ, Basnett P, Lukasiewicz B, Stevens M, Best S, Cameron R, Sinha S, Harding SE, Roy I. Natural Biomaterials for Cardiac Tissue Engineering: A Highly Biocompatible Solution. Front Cardiovasc Med. 2020 Oct 23;7:554597