Repertorio de linfocitos T y perfiles de expresión génica y mutacional: nuevos biomarcadores en quimio- inmunoterapia neoadyuvante de cáncer de pulmón

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de Lectura: 01-09-2022El cáncer de pulmón no microcítico representa el 80% de los casos de cáncer de pulmón, y un tercio de los pacientes son diagnosticados con un estadio localmente avanzado considerado potencialmente curable y resecable. El tratamiento neoadyuvante busca la reducción del tamaño tumoral para mejorar su resecabilidad además de tratar de manera temprana la enfermedad metastásica. En el ensayo clínico NADIM se presenta un nuevo enfoque terapéutico con la adición de la inmunoterapia a la quimioterapia en el tratamiento neoadyuvante de pacientes con cáncer de pulmón no microcítico en estadios IIIA. Las altas tasas de respuesta patológica completa (CPR) logradas en este ensayo han dado lugar a la búsqueda de biomarcadores que ayuden en la predicción de la respuesta a la intervención. Por tanto, el objetivo principal de este estudio es la identificación de biomarcadores de respuesta a la quimio-inmunoterapia neoadyuvante, así como la caracterización del microambiente tumoral. Para ello, se ha llevado a cabo la determinación de biomarcadores utilizados previamente como es la expresión de PD-L1, así como la determinación de la carga mutacional tumoral en las muestras de tejido basal. Además, proponemos biomarcadores emergentes como son el análisis mediante tecnología NGS del receptor de linfocitos T (TCR), y la expresión génica de una firma inmunitaria en las muestras de tejido pre- y postratamiento. En este trabajo se muestra cómo una mayor expresión de PD-L1 en el tejido pretratamiento se asocia positivamente a la respuesta al tratamiento con inmunoterapia. Además, se ha encontrado una asociación positiva entre un repertorio de TCR más sesgado en las muestras pretratamiento y la CPR. El repertorio de TCR de los tumores CPR en el momento pretratamiento parece estar dominado por un conjunto de clones más frecuentes (top 1%) que ocupa un mayor espacio clonal con respecto al total y que caracteriza a los tumores que responderán de manera completa al tratamiento neoadyuvante. Por otro lado, la firma de expresión génica caracteriza los tumores CPR en el momento pretratamiento, mostrando estos un perfil proinflamatorio dominado por una firma génica de IFNγ. Asimismo, los mayores cambios en la expresión génica producidos tras el tratamiento neoadyuvante se encuentran en los tumores CPR, reflejando una respuesta inmunitaria antitumoral activa que cesa tras la resolución del tumor. En conclusión, los resultados reflejan las diferencias entre las respuestas completas y no completas y le dan valor a la respuesta patológica completa como una entidad con características inmunitarias diferenciadasEste trabajo ha sido posible gracias a la financiación de Bristol-Myers Squibb, Thermo Fisher Scientific, el Instituto de Salud Carlos II (PI19/01652) y Horizonte 202

Terapia regenerativa celular en el tratamiento de la Esclerosis Múltiple

Con la terapia regenerativa se busca la curación completa del paciente que presenta una determinada patología y conociendo el mecanismo por el cual se produce dicha enfermedad, es posible acercarse a su solución. En este caso, se estudia el potencial de las células madre mesenquimales para revertir la esclerosis múltiple. Además, se propone la encapsulación de las células como método de liberación de las mismas al interior del organismo, favoreciendo su potencial terapéutico

The role of metabolism in tumor immune evasion: Novel approaches to improve immunotherapy

The tumor microenvironment exhibits altered metabolic properties as a consequence of the needs of tumor cells, the natural selection of the most adapted clones, and the selfish relationship with other cell types. Beyond its role in supporting uncontrolled tumor growth, through energy and building materials obtention, metabolism is a key element controlling tumor immune evasion. Immunotherapy has revolutionized the treatment of cancer, being the first line of treatment for multiple types of malignancies. However, many patients either do not benefit from immunotherapy or eventually relapse. In this review we overview the immunoediting process with a focus on the metabolism-related elements that are responsible for increased immune evasion, either through reduced immunogenicity or increased resistance of tumor cells to the apoptotic action of immune cells. Finally, we describe the main molecules to modulate these immune evasion processes through the control of the metabolic microenvironment as well as their clinical developmental statusWork in the authors’ laboratories was supported by “Instituto de Salud Carlos III” (ISCIII) PI19/01652 grant, Ministry of Science and Innovation RTC2017-6502-1 INmunoSIGHT and European Union’s Horizon 2020 research and innovation programme, CLARIFY 875160 grant, to M.P. A.C.-B. received a Spanish Lung Cancer Group (SLCG) grant and is supported by a ISCIII-“Sara Borrell” contract CD19/00170. M.C. is supported by PEJD-2019-PRE/BMD-17006 contract granted to A.C.-B. R.L.-B. was supported by PEJ16/MED/AI-1972 and PEJD-2018-PRE/SAL-8641 from European Social Fund and Comunidad de Madrid, both granted to M.

Tumor microenvironment gene expression profiles associated to complete pathological response and disease progression in resectable NSCLC patients treated with neoadjuvant chemoimmunotherapy

Gene expression profiling; Lung neoplasms; Tumor biomarkersPerfil d'expressió gènica; Neoplàsies pulmonars; Biomarcadors tumoralsPerfil de expresión génica; Neoplasias pulmonares; Biomarcadores tumoralesBackground Neoadjuvant chemoimmunotherapy for non-small cell lung cancer (NSCLC) has improved pathological responses and survival rates compared with chemotherapy alone, leading to Food and Drug Administration (FDA) approval of nivolumab plus chemotherapy for resectable stage IB-IIIA NSCLC (AJCC 7th edition) without ALK or EGFR alterations. Unfortunately, a considerable percentage of tumors do not completely respond to therapy, which has been associated with early disease progression. So far, it is impossible to predict these events due to lack of knowledge. In this study, we characterized the gene expression profile of tumor samples to identify new biomarkers and mechanisms behind tumor responses to neoadjuvant chemoimmunotherapy and disease recurrence after surgery. Methods Tumor bulk RNA sequencing was performed in 16 pretreatment and 36 post-treatment tissue samples from 41 patients with resectable stage IIIA NSCLC treated with neoadjuvant chemoimmunotherapy from NADIM trial. A panel targeting 395 genes related to immunological processes was used. Tumors were classified as complete pathological response (CPR) and non-CPR, based on the total absence of viable tumor cells in tumor bed and lymph nodes tested at surgery. Differential-expressed genes between groups and pathway enrichment analysis were assessed using DESeq2 and gene set enrichment analysis. CIBERSORTx was used to estimate the proportions of immune cell subtypes. Results CPR tumors had a stronger pre-established immune infiltrate at baseline than non-CPR, characterized by higher levels of IFNG, GZMB, NKG7, and M1 macrophages, all with a significant area under the receiver operating characteristic curve (ROC) >0.9 for CPR prediction. A greater effect of neoadjuvant therapy was also seen in CPR tumors with a reduction of tumor markers and IFNγ signaling after treatment. Additionally, the higher expression of several genes, including AKT1, BST2, OAS3, or CD8B; or higher dendritic cells and neutrophils proportions in post-treatment non-CPR samples, were associated with relapse after surgery. Also, high pretreatment PD-L1 and tumor mutational burden levels influenced the post-treatment immune landscape with the downregulation of proliferation markers and type I interferon signaling molecules in surgery samples. Conclusions Our results reinforce the differences between CPR and non-CPR responses, describing possible response and relapse immune mechanisms, opening the possibility of therapy personalization of immunotherapy-based regimens in the neoadjuvant setting of NSCLC.Work in the authors’ laboratories was supported by '‘Instituto de Salud Carlos III’' (ISCIII) PI19/01652 grant cofunded by European Regional Development Fund (ERDF), Bristol-Myers Squibb (BMS), Ministry of Science and Innovation RTC2017-6502-1 'INmunoSIGHT', RTC2019-007359-1 'BLI-O' and European Union’s Horizon 2020 research and innovation programme, CLARIFY 875160 grant, to MP. ThermoFisher provided reagents for RNA sequencing. AC-B received a Spanish Lung Cancer Group (SLCG) grant and is supported by a ISCIII-“Sara Borrell” contract CD19/00170. MCa is supported by PEJD-2019-PRE/BMD-17006 contract granted to AC-B

Pretreatment Tissue TCR Repertoire Evenness Is Associated with Complete Pathologic Response in Patients with NSCLC Receiving Neoadjuvant Chemoimmunotherapy

Neoadjuvant Chemoimmunotherapy; Lung cancer; T-cell receptorQuimioimmunoteràpia neoadjuvant; Càncer de pulmó; Receptor de cèl·lules TQuimioinmunoterapia neoadyuvante; Cáncer de pulmón; Receptor de células TPurpose: Characterization of the T-cell receptor (TCR) repertoire may be a promising source for predictive biomarkers of pathologic response to immunotherapy in locally advanced non–small cell lung cancer (NSCLC). Experimental Design: In this study, next-generation TCR sequencing was performed in peripheral blood and tissue samples of 40 patients with NSCLC, before and after neoadjuvant chemoimmunotherapy (NADIM clinical trial, NCT03081689), considering their complete pathologic response (CPR) or non-CPR. Beyond TCR metrics, tissue clones were ranked by their frequency and spatiotemporal evolution of top 1% clones was determined. Results: We have found a positive association between an uneven TCR repertoire in tissue samples at diagnosis and CPR at surgery. Moreover, TCR most frequently ranked clones (top 1%) present in diagnostic biopsies occupied greater frequency in the total clonal space of CPR patients, achieving an AUC ROC to identify CPR patients of 0.967 (95% confidence interval, 0.897–1.000; P = 0.001), and improving the results of PD-L1 tumor proportion score (TPS; AUC = 0.767; P = 0.026) or tumor mutational burden (TMB; AUC = 0.550; P = 0.687). Furthermore, tumors with high pretreatment top 1% clonal space showed similar immune cell populations but a higher immune reactive gene expression profile. Finally, the selective expansion of pretreatment tissue top 1% clones in peripheral blood of CPR patients suggests also a peripheral immunosurveillance, which could explain the high survival rate of these patients. Conclusions: We have identified two parameters derived from TCR repertoire analysis that could outperform PD-L1 TPS and TMB as predictive biomarkers of CPR after neoadjuvant chemoimmunotherapy, and unraveled possible mechanisms of CPR involving enhanced tumor immunogenicity and peripheral immunosurveillance

Hacia la creación de una colección de referencia de la fauna española de kinorrincos (Scalidophora: Kinorhyncha) en el Museo Nacional de Ciencias Naturales de Madrid (MNCN-CSIC)

The Invertebrate Collection is part of the 17 collections from the National Museum of Natural Science of Madrid (MNCN-CSIC). Up to now, this collection included specimens of 27 animal phyla. The present contribution determines the donation of 44 specimens belonging to 25 species, 13 genera and six families of the phylum Kinorhyncha, increasing the number of represented phyla of the aforementioned collection to 28. For each species, locality, date and project of origin are provided. Furthermore, the species Pycnophyes giganteus is reported for the first time in Balearic waters.La Colección de Invertebrados es una de las 17 colecciones científicas del Museo Nacional de Ciencias Naturales de Madrid (MNCN-CSIC). Hasta la fecha, dicha colección comprendía ejemplares pertenecientes a 27 filos animales. El presente trabajo informa de la donación de 44 ejemplares pertenecientes a 25 especies, 13 géneros y seis familias del filo Kinorhyncha, lo que amplía hasta 28 el número de filos representados en la citada colección. Para cada especie se aportan datos del lugar, fecha y proyecto de procedencia. Además, se reporta por primera vez la especie Pycnophyes giganteus en aguas baleares

Tumor microenvironment gene expression profiles associated to complete pathological response and disease progression in resectable NSCLC patients treated with neoadjuvant chemoimmunotherapy

Background Neoadjuvant chemoimmunotherapy for non-small cell lung cancer (NSCLC) has improved pathological responses and survival rates compared with chemotherapy alone, leading to Food and Drug Administration (FDA) approval of nivolumab plus chemotherapy for resectable stage IB-IIIA NSCLC (AJCC 7th edition) without ALK or EGFR alterations. Unfortunately, a considerable percentage of tumors do not completely respond to therapy, which has been associated with early disease progression. So far, it is impossible to predict these events due to lack of knowledge. In this study, we characterized the gene expression profile of tumor samples to identify new biomarkers and mechanisms behind tumor responses to neoadjuvant chemoimmunotherapy and disease recurrence after surgery. Methods Tumor bulk RNA sequencing was performed in 16 pretreatment and 36 post-treatment tissue samples from 41 patients with resectable stage IIIA NSCLC treated with neoadjuvant chemoimmunotherapy from NADIM trial. A panel targeting 395 genes related to immunological processes was used. Tumors were classified as complete pathological response (CPR) and non-CPR, based on the total absence of viable tumor cells in tumor bed and lymph nodes tested at surgery. Differential-expressed genes between groups and pathway enrichment analysis were assessed using DESeq2 and gene set enrichment analysis. CIBERSORTx was used to estimate the proportions of immune cell subtypes. Results CPR tumors had a stronger pre-established immune infiltrate at baseline than non-CPR, characterized by higher levels of IFNG, GZMB, NKG7, and M1 macrophages, all with a significant area under the receiver operating characteristic curve (ROC) >0.9 for CPR prediction. A greater effect of neoadjuvant therapy was also seen in CPR tumors with a reduction of tumor markers and IFN gamma signaling after treatment. Additionally, the higher expression of several genes, including AKT1, BST2, OAS3, or CD8B; or higher dendritic cells and neutrophils proportions in post-treatment non-CPR samples, were associated with relapse after surgery. Also, high pretreatment PD-L1 and tumor mutational burden levels influenced the post-treatment immune landscape with the downregulation of proliferation markers and type I interferon signaling molecules in surgery samples. Conclusions Our results reinforce the differences between CPR and non-CPR responses, describing possible response and relapse immune mechanisms, opening the possibility of therapy personalization of immunotherapy-based regimens in the neoadjuvant setting of NSCLC

Adquisición de competencias profesionales en los estudiantes del Grado de Trabajo Social. La argumentación y elaboración en el diagnóstico social a través de diferentes instrumentos

Se presentan los resultados obtenidos en el Proyecto Nº 21 Adquisición de competencias profesionales en los estudiantes del Grado de Trabajo Social. La argumentación y elaboración en el diagnóstico social a través de diferentes instrumentos, acerca de reforzar la argumentación en el alumnado del Grado de Trabajo Social a través de instrumentos, que refuercen su aprendizaje en la elaboración del diagnóstico social, donde se visualiza la calidad y eficacia de la intervención profesional

The genus setaphyes (kinorhyncha, pycnophyidae) in european waters: Redescription of Setaphyes dentatus ( ) and Setaphyes kielensis ( ), including notes on morphometrics, sexually dimorphic features and reproduction of the genus

Six of the eight species of the genus Setaphyes have been described from Europe. In the present contribution, the European Setaphyes species are revised and two of them, S. dentatus (Reinhard, 1881) and S. kielensis (Zelinka, 1928), are redescribed following the current approaches of Kinorhyncha taxonomy. For this purpose, material from 17 localities throughout the Atlantic Ocean and the Mediterranean and Black Seas was investigated. The redescriptions of two of the most common pycnophyid species in Europe include detailed information on intraspecific variation among populations. Setaphyes dentatus is characterized by the presence of middorsal elevations on segments 2–6, and middorsal processes on segments 1, 7–9, unpaired paradorsal setae on segments 2–9; laterodorsal setae on segments 3, 5 and 7; ventromedial setae on segments 4–5, 7–8 in males and 3–5, 7–9 in females; anterior cuticular, reticulate ridges on the tergal plate of segment 1 and conspicuous patch of laterodorsal and ventrolateral longitudinal, cuticular ridges on segment 10. Setaphyes kielensis is characterized by middorsal elevations on segments 1–9; paired paradorsal setae on segments 2–9 (unpaired on segment 8); laterodorsal setae on segments 2–9; ventromedial setae on segments 3–9. Furthermore, morphometric features of the European Setaphyes are analysed. Specifically, the morphometric analyses revealed sex-specific variability in the LTS/TL proportion, with females of all the European species of Setaphyes showing smaller LTS/TL. Thus, the LTS/TL ratio unveils a conspicuous sexual dimorphism in the genus

The genus setaphyes (kinorhyncha, pycnophyidae) in european waters: Redescription of Setaphyes dentatus (Reinhard, 1881) and Setaphyes kielensis (Zelinka, 1928), including notes on morphometrics, sexually dimorphic features and reproduction of the genus

Six of the eight species of the genus Setaphyes have been described from Europe. In the present contribution, the European Setaphyes species are revised and two of them, S. dentatus (Reinhard, 1881) and S. kielensis (Zelinka, 1928), are redescribed following the current approaches of Kinorhyncha taxonomy. For this purpose, material from 17 localities throughout the Atlantic Ocean and the Mediterranean and Black Seas was investigated. The redescriptions of two of the most common pycnophyid species in Europe include detailed information on intraspecific variation among populations. Setaphyes dentatus is characterized by the presence of middorsal elevations on segments 2–6, and middorsal processes on segments 1, 7–9, unpaired paradorsal setae on segments 2–9; laterodorsal setae on segments 3, 5 and 7; ventromedial setae on segments 4–5, 7–8 in males and 3–5, 7–9 in females; anterior cuticular, reticulate ridges on the tergal plate of segment 1 and conspicuous patch of laterodorsal and ventrolateral longitudinal, cuticular ridges on segment 10. Setaphyes kielensis is characterized by middorsal elevations on segments 1–9; paired paradorsal setae on segments 2–9 (unpaired on segment 8); laterodorsal setae on segments 2–9; ventromedial setae on segments 3–9. Furthermore, morphometric features of the European Setaphyes are analysed. Specifically, the morphometric analyses revealed sex-specific variability in the LTS/TL proportion, with females of all the European species of Setaphyes showing smaller LTS/TL. Thus, the LTS/TL ratio unveils a conspicuous sexual dimorphism in the genus