Quantification of cells expressing mesenchymal stem cell markers in healthy and osteoarthritic synovial membranes

[Abstract] Objective. To quantify cells expressing mesenchymal stem cell (MSC) markers in synovial mem- branes from human osteoarthritic (OA) and healthy joints. Methods. Synovial membranes from OA and healthy joints were digested with collagenase and the isolated cells were cultured. Synovial membrane-derived cells were phenotypically characterized for differentiation experiments using flow cytometry to detect the expression of mesenchymal markers (CD29, CD44, CD73, CD90, CD105, CD117, CD166, and STRO-1) and hematopoietic markers (CD34 and CD45). Chondrogenesis was assessed by staining for proteoglycans and collagen type II, adipogenesis by using a stain for lipids, and osteogenesis by detecting calcium deposits. Coexpression of CD44, CD73, CD90, and CD105 was determined using immunofluorescence. Results. Cells expressing MSC markers were diffusely distributed in OA synovial membranes; in healthy synovial membrane these cells were localized in the subintimal zone. More numerous MSC markers in OA synovial membranes were observed in cells also expressing the CD90 antigen. FACS analysis showed that more than 90% of OA synovial membrane-derived cells were positive for CD44, CD73, and CD90, and negative for CD34 and CD45. OA synovial membrane-derived cells were also positive for CD29 (85.23%), CD117 (72.35%), CD105 (45.5%), and STRO-1 (49.46%). Micropellet analyses showed that the culture of cells with transforming growth factor-ß3 stimulated proteoglycan and collagen type II synthesis. Conclusion. Synovial membranes from patients with OA contain more cells positive for CD44, CD90, and CD105 antigens than those from joints with undamaged cartilage

Bone marrow cells immunomagnetically selected for CD271+ antigen promote in vitro the repair of articular cartilage defects

[Abstract] Objective: The purposes of this project were to quantify the cells expressing the mesenchymal stem cell (MSC) marker CD271 in synovial membranes from human osteoarthritic (OA) and healthy joints, and to determine if those CD271 cells were involved in spontaneous human cartilage repair and were beneficial for the repair of human articular cartilage defects. Methods: The coexpression of CD44/CD271, CD90/CD271, and CD105/CD271 antigens was determined by immunofluorescence in OA and healthy synovial membranes and during spontaneous cartilage repair. Isolated MSCs from the bone marrow of four OA patients (mean age: 64 years) were magnetically separated into MSC CD271+ and MSC CD271 subsets. The separated cell subsets were then implanted into 2 mm focal defects of articular cartilage. These implants were cultured in chondrogenic differentiation medium supplemented with recombinant human transforming growth factor-beta3 for 8 weeks. The repair tissues were analyzed by histochemistry (hematoxylin–eosin and safranin O) and immunohistochemistry for collage types I and II. Results: Cells expressing the CD271 antigen were diffusely distributed in OA synovial membranes and localized in the subintimal zone in healthy synovial membranes. The number of cells expressing MSC markers was higher in OA synovial membranes than in synovia from healthy joints, corresponding to the highest level of coexpression of CD90/CD271 antigens (9.8% vs. 2.6%). Spontaneous repair tissue showed more cells expressing the CD271 antigen (9.9% ± 4.0%). The highest levels of expression were found to be associated with CD44; 64% of positive CD271 cells coexpressed the CD44 antigen. In both implant cell types, the repair tissue morphology resembled articular cartilage, having an extracellular matrix with a hyaline aspect and numerous lacunae containing cells, and was immunopositive for collagen types I and II. Statistical analyses of the repair tissue demonstrated that the implantation of MSC CD271+ provided such benefits as a greater filling of the chondral defect and better integration between the repair tissue and native cartilage. Safranin O staining of repair tissue was negative in implants of MSC CD271- but more positive in implants with MSC CD271+. The overall histologic score for CD271 implants was 9.5 ± 0.89 and 12.19 ± 1.01 for CD271+ implants. Conclusions: Synovial membranes from OA patients contain more cells expressing CD271 antigen than those from healthy joints, and spontaneous cartilage repair tissue contains cells positive for CD271 antigen. These data suggest the involvement of CD271 antigen in spontaneous cartilage repair and indicate that the cell subset MSC CD271+ provides higher quality chondral repair than the CD271- subset.Galicia. Consellería de Economía, Industria e Comercio; PGIDIT05SAN08PRGalicia. Consellería de Economía, Industria e Comercio; PGIDIT04PXIC91602PNGalicia. Consellería de Economía, Industria e Comercio; PGIDIT04PXIC91601PNInstituto de Salud Carlos III; FISCP03/00127Instituto de Salud Carlos III; FIS05/249

YES1 drives lung cancer growth and progression and predicts sensitivity to dasatinib

Rationale: The characterization of new genetic alterations is essential to assign effective personalized therapies in non–small cell lung cancer (NSCLC). Furthermore, finding stratification biomarkers is essential for successful personalized therapies. Molecular alterations of YES1, a member of the SRC (proto-oncogene tyrosine-protein kinase Src) family kinases (SFKs), can be found in a significant subset of patients with lung cancer. Objectives: To evaluate YES1 (v-YES-1 Yamaguchi sarcoma viral oncogene homolog 1) genetic alteration as a therapeutic target and predictive biomarker of response to dasatinib in NSCLC. Methods: Functional significance was evaluated by in vivo models of NSCLC and metastasis and patient-derived xenografts. The efficacy of pharmacological and genetic (CRISPR [clustered regularly interspaced short palindromic repeats]/Cas9 [CRISPR-associated protein 9]) YES1 abrogation was also evaluated. In vitro functional assays for signaling, survival, and invasion were also performed. The association between YES1 alterations and prognosis was evaluated in clinical samples. Measurements and Main Results: We demonstrated that YES1 is essential for NSCLC carcinogenesis. Furthermore, YES1 overexpression induced metastatic spread in preclinical in vivo models. YES1 genetic depletion by CRISPR/Cas9 technology significantly reduced tumor growth and metastasis. YES1 effects were mainly driven by mTOR (mammalian target of rapamycin) signaling. Interestingly, cell lines and patient-derived xenograft models with YES1 gene amplifications presented a high sensitivity to dasatinib, an SFK inhibitor, pointing out YES1 status as a stratification biomarker for dasatinib response. Moreover, high YES1 protein expression was an independent predictor for poor prognosis in patients with lung cancer. Conclusions: YES1 is a promising therapeutic target in lung cancer. Our results provide support for the clinical evaluation of dasatinib treatment in a selected subset of patients using YES1 status as predictive biomarker for therapy

Objetivos de desarrollo sostenible y Derechos Humanos: paz, justicia e instituciones sólidas / Derechos Humanos y empresas

Nota previa / Cástor Miguel Díaz Barrado (pp. 7-9). -- Unión Europea: Derechos Humanos y desarrollo sostenible / Araceli Mangas Martín (pp. 13 - 26). -- Paz, seguridad y gobernanza: el ODS 16 y la Agenda 2030 de desarrollo sostenible / José Antonio Sanahuja (pp. 27 - 54). -- La aplicación extraterritorial de los Derechos Humanos por acciones de empresas / Pablo Antonio Fernández Sánchez (pp. 57 - 60). -- Hacia un futuro tratado internacional sobre las empresas y los Derechos Humanos / Eugenia López-Jacoiste (pp. 61 - 73). -- La importancia del enfoque de los Derechos Humanos en los objetivos de desarrollo del sostenible / Diana M. Verdiales López (pp. 75 - 90). -- La perspectiva de género en la resolución de conflictos armados: las mujeres como agentes de paz / Cristina del Prado Higuera (pp. 93 - 106). -- La mutilación genital femenina desde una perspectiva integral y multidisciplinar / Cristina Hermida del Llano (pp. 107 - 120). -- Poner fin al maltrato, la explotación, la trata y todas las formas de violencia y tortura contra los niños: la trata de niños / María Ángeles Cano Linares (pp. 123 - 136). -- Participación juvenil en el fortalecimiento de las instituciones y el establecimiento de la paz: programas de participación juvenil del sistema de Naciones Unidas / Enrique Hernández Díez (pp. 137 - 160). -- Estado de la violencia criminal en la sociedad internacional: respuestas de la comunidad internacional para avanzar hacia el objetivo 16, Agenda 2030 / Sagrario Morán Blanco (pp. 163 - 178). -- Fortalecer la recuperación y devolución de bienes robados: justicia y reparación del daño causado a las víctimas y la sociedad como metas del objetivo 16 de los objetivos de desarrollo sostenible / Francisco Jiménez García (pp. 179 - 192). -- El derecho internacional, los ODS y la lucha contra el crimen transnacional / Juan Manuel Rodríguez Barrigón (pp. 193 - 216). -- El derecho internacional frente a la criminalidad financiera transnacional: la prevención y sanción del blanqueo de capitales / Jorge Urbaneja Cillán (pp. 217 - 232). -- Acceso a la justicia y objetivos del desarrollo sostenible / Florabel Quispe Remón (pp. 235 - 248). -- Paz y objetivos de desarrollo sostenible: la contribución del objetivo 16 / Elena C. Díaz Galán (pp. 249 - 262). -- Seguridad sanitaria y empresas / Ana Cristina Gallego Hernández (pp. 265 - 272). -- El acaparamiento de tierras por empresas multinacionales y la seguridad alimentaria / Adriana Fillol Mazo (pp. 273 - 288). -- Importancia y características de los planes nacionales de empresas y Derechos Humanos en relación con el desarrollo sostenible / Alberto Jiménez-Piernas García (pp. 289 - 301). -- Objetivo de desarrollo sostenible 16. Principios rectores y espacio iberoamericano: el caso Berta Cáceres / Ana Manero Salvador (pp. 305 - 315). -- Derechos de los pueblos indígenas: marcos de protección en la Agenda 2030 y en los principios rectores sobre empresas y Derechos Humanos / Juan Daniel Oliva Martínez y Adriana Sánchez Lizama (pp. 317 - 331

Identification of novel biomarkers and therapeutic targets for diagnosis and clinical managment of patients with head and neck squamour cell carcinomas

Tesis con mención internacionalEl objetivo global de esta tesis ha consistido en investigar alteraciones genéticas/moleculares en carcinomas epidermoides de cabeza y cuello (CECC) y su potencial aplicación clínica como marcadores de diagnóstico, pronóstico y/o nuevas dianas terapéuticas. Hemos demostrado la relevancia clínica de cortactina (CTTN) y la quinasa de adhesión focal (FAK) como potentes predictores de riesgo de cáncer de laringe, superiores a la clasificación histológica, actual estándar en la práctica clínica. Además el bloqueo farmacológico o funcional de FAK inhibe eficazmente el crecimiento celular, la transformación in vitro y la invasión. Esta tesis también demuestra el impacto diferencial de la expresión de la quinasa Src en la supervivencia de pacientes con CECC dependiendo de la localización del tumor y, más importante, revela sorprendentes efectos nocivos de los inhibidores de Src dasatinib y saracatinib promoviendo las propiedades stem, lo cual proporciona una posible explicación para la falta de eficacia clínica de ambos fármacos en pacientes con CECC. Finalmente, un nuevo método de screening de genoma completo utilizando el sistema CRISPR/Cas9 ha permitido identificar un número de genes esenciales y susceptibilidades genéticas en CECC, que podrían constituir la base para el desarrollo de estrategias terapéuticas personalizadas