Nanomecánica del celulosoma. Implicaciones para la actividad del sistema

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 05-02-2016Esta tesis tiene embargado el acceso al texto completo hasta el 05-02-2017Cellulose is the major biopolymer and carbon source on the biosphere. It is composed of glucose, a sugar that can be processed by several microorganisms to produce added-value chemicals such as biofuels. The major bottleneck towards its utilization in industry is the saccharification step, i. e. the breakdown of the polymer to release the constituent soluble sugars. In nature, this process is carried out by several microorganisms with specialized enzymatic machinery. In particular, some anaerobic bacteria have developed a high molecular weight complex known as the cellulosome. This comprises a large usually non-catalytic protein called scaffoldin, capable of docking several enzymes and targeting them to the cellulosic substrate while binding it to the cell surface. As an adhesion system, we expect that the cellulosome is subjected to mechanical stress since the relative movement of the cell and its substrate would stretch the connecting region: the portion of the scaffoldin located between the two anchoring points. We hypothesized that the mechanical stability, i. e. the resistance to forced unfolding, of this region might be important to understand cellulosome activity since the unfolding of cohesins, which are the scaffoldin modules capable of binding enzymes, might result in enzyme release and activity loss. We have used atomic force microscopy-based single molecule force spectroscopy in combination with molecular dynamics simulations to study the mechanical properties of several cohesins from different cellulosomes. We found that those cohesins located in the connecting region of scaffoldins that bind enzymes are more mechanically stable than those located outside the two anchoring points, in the external region. This principle applies for cohesins of different cellulosomes despite the sequence difference of these modules. We also study the effects of both the linker sequences located between cohesins and enzyme binding on the mechanical stability of cohesins which are elements that are found in natural cellulosomes. We did not observe any effect on cohesin mechanical stability nor on their mechanical clamp positionLa celulosa es el biopolímero y la fuente de carbono más abundante de la biosfera. Está compuesta por glucosa, un azúcar que puede ser procesado por microorganismos para producir productos químicos con valor añadido como los biocombustibles. El mayor cuello de botella para su utilización industrial se encuentra en el paso de la sacarificación, es decir, la degradación de este polímero para liberar los azúcares solubles constituyentes. En la naturaleza este proceso lo llevan a cabo algunos microorganismos que presentan una maquinaria enzimática especializada. En particular, algunas bacterias anaerobias han desarrollado un complejo conocido como celulosoma que comprende una proteína de elevado peso molecular y generalmente no catalítica llamada escafoldina, capaz de unir varias enzimas y dirigirlas al sustrato a la vez que las ancla a la superficie celular. Por el hecho de tratarse de un sistema de adhesión, es de esperar que el celulosoma esté sometido a estrés mecánico dado que el movimiento relativo de la bacteria respecto al sustrato podría estirar la región conectora: la porción de la escafoldina que se encuentra entre los dos puntos de anclaje. Propusimos la hipótesis de que la estabilidad mecánica, es decir, la resistencia al desplegamiento por fuerza, de esta región puede ser importante para entender la actividad del celulosoma. Esto es debido a que el desplegamiento de las cohesinas, los módulos de la escafoldina capaces de unir enzimas, podría conducir a una pérdida de actividad. Hemos empleado espectroscopia de fuerza de moléculas individuales mediante microscopía de fuerza atómica en combinación con simulaciones de dinámica molecular para estudiar las propiedades mecánicas de varios módulos cohesina de distintos celulosomas. Hemos observado que las cohesinas situadas en la región conectora presentan una mayor estabilidad mecánica comparada con la de aquellas que no se espera que estén sometidas a estrés mecánico. Este principio es aplicable a las cohesinas de distintos celulosomas a pesar de las diferencias de secuencia entre ellas. También hemos estudiado el efecto de las secuencias intermodulares y de la unión a enzimas sobre la estabilidad mecánica de las cohesinas, que son elementos que se encuentran en los celulosomas naturales. Hemos visto que ni la estabilidad mecánica de las cohesinas ni la posición de su parche mecánico se ven afectados por estos factore

Estudio semémico del adjetivo «bueno» en «Rinconete y cortadillo» de Cervantes

El estudio de cada una de las ocurrencias de bueno en las diferentes lexías de las que forma parte en el texto de Cervantes, nos dará su valor semémico

Nanomechanics of tip-link cadherins

Hearing and balance rely on the transduction of mechanical stimuli arising from sound waves or head movements into electrochemical signals. This archetypal mechanoelectrical transduction process occurs in the hair-cell stereocilia of the inner ear, which experience continuous oscillations driven by undulations in the endolymph in which they are immersed. The filamentous structures called tip links, formed by an intertwined thread composed of an heterotypic complex of cadherin 23 and protocadherin 15 ectodomain dimers, connect each stereocilium to the tip of the lower sterocilium, and must maintain their integrity against continuous stimulatory deflections. By using single molecule force spectroscopy, here we demonstrate that in contrast to the case of classical cadherins, tip-link cadherins are mechanoresilient structures even at the exceptionally low Ca2+ concentration of the endolymph. We also show that the D101G deafness point mutation in cadherin 23, which affects a Ca2+ coordination site, exhibits an altered mechanical phenotype at the physiological Ca2+ concentration. Our results show a remarkable case of functional adaptation of a protein’s nanomechanics to extremely low Ca2+ concentrations and pave the way to a full understanding of the mechanotransduction mechanism mediated by auditory cadherinsThis work was supported by the BIO2010-22275 grant from the Spanish Ministry of Science and Innovation (MICINN) to M.C.-V

ICU-acquired pneumonia is associated with poor health post-COVID-19 syndrome

Some patients previously presenting with COVID-19 have been reported to develop persistent COVID-19 symptoms. While this information has been adequately recognised and extensively published with respect to non-critically ill patients, less is known about the incidence and factors associated with the characteristics of persistent COVID-19. On the other hand, these patients very often have intensive care unit-acquired pneumonia (ICUAP). A second infectious hit after COVID increases the length of ICU stay and mechanical ventilation and could have an influence on poor health post-COVID 19 syndrome in ICU-discharged patients. Methods: This prospective, multicentre, and observational study was carrid out across 40 selected ICUs in Spain. Consecutive patients with COVID-19 requiring ICU admission were recruited and evaluated three months after hospital discharge. Results: A total of 1255 ICU patients were scheduled to be followed up at 3 months; however, the final cohort comprised 991 (78.9%) patients. A total of 315 patients developed ICUAP (97% of them had ventilated ICUAP). Patients requiring invasive mechanical ventilation had more persistent post-COVID-19 symptoms than those who did not require mechanical ventilation. Female sex, duration of ICU stay, development of ICUAP, and ARDS were independent factors for persistent poor health post-COVID-19. Conclusions: Persistent post-COVID-19 symptoms occurred in more than two-thirds of patients. Female sex, duration of ICU stay, development of ICUAP, and ARDS all comprised independent factors for persistent poor health post-COVID-19. Prevention of ICUAP could have beneficial effects in poor health post-COVID-19.Financial support was provided by the Instituto Carlos III de Madrid (COV20/00110, ISCIII) and by the Centro de Investigación Biomedica En Red—Enfermedades Respiratorias (CIBERES). DdGC has received financial support from Instituto de Salud Carlos III (Miguel Servet 2020: CP20/00041), co-funded by European Social Fund (ESF)/ “Investing in your future”Peer ReviewedArticle signat per 53 autors/es: Ignacio Martin-Loeches (1,2,3), Anna Motos (1,3), Rosario Menéndez (1,4), Albert Gabarrús (1,4), Jessica González (5,6), Laia Fernández-Barat (1,3), Adrián Ceccato (1,3), Raquel Pérez-Arnal (7), Dario García-Gasulla (7), Ricard Ferrer (1,8), Jordi Riera (1,8), José Ángel Lorente (1,9), Óscar Peñuelas (1,9), Jesús F. Bermejo-Martin (1,10,11), David de Gonzalo-Calvo (5,6), Alejandro Rodríguez (12), Ferran Barbé (5,6), Luciano Aguilera (13), Rosario Amaya-Villar (14), Carme Barberà (15), José Barberán (16), Aaron Blandino Ortiz (17), Elena Bustamante-Munguira (18), Jesús Caballero (19), Cristina Carbajales (20), Nieves Carbonell (21),Mercedes Catalán-González (22), Cristóbal Galbán (23), Víctor D. Gumucio-Sanguino (24), Maria del Carmen de la Torre (25), Emili Díaz (26), Elena Gallego (27), José Luis García Garmendia (28), José Garnacho-Montero (29), José M. Gómez (30), Ruth Noemí Jorge García (31), Ana Loza-Vázquez (32), Judith Marín-Corral (33), Amalia Martínez de la Gándara (34), Ignacio Martínez Varela (35), Juan Lopez Messa (36), Guillermo M. Albaiceta (37,38), Mariana Andrea Novo (39), Yhivian Peñasco (40), Pilar Ricart (41), Luis Urrelo-Cerrón (42), Angel Sánchez-Miralles (43), Susana Sancho Chinesta (44), Lorenzo Socias (45), Jordi Solé-Violan (1,46), Luis Tamayo Lomas (47), Pablo Vidal (48) and Antoni Torres (1,3)*, on behalf of CIBERESUCICOVID Project (COV20/00110 and ISCIII) // (1) CIBER of Respiratory Diseases (CIBERES), Institute of Health Carlos III, 28029 Madrid, Spain; (2) Pulmonary Department, Hospital Clinic, Universitat de Barcelona, IDIBAPS, 08036 Barcelona, Spain; (3) Department of Intensive Care Medicine, St. James’s Hospital, Multidisciplinary Intensive Care Research Organization (MICRO), James’s Street, D08 NHY1 Dublin, Ireland; (4) Pulmonary Department, University and Polytechnic Hospital La Fe, 46026 Valencia, Spain; (5) Translational Research in Respiratory Medicine Group (TRRM), Lleida Biomedical Research Institute (IRBLleida), 25198 Lleida, Spain; (6) Pulmonary Department, Hospital Universitari Arnau de Vilanova and Santa Maria, 25198 Lleida, Spain; (7) Barcelona Supercomputing Centre (BSC), 08034 Barcelona, Spain; (8) Intensive Care Department, Vall d’Hebron Hospital Universitari, SODIR Research Group, Vall d’Hebron Institut de Recerca (VHIR), 08035 Barcelona, Spain; (9) Hospital Universitario de Getafe, 28905 Madrid, Spain; (10) Hospital Universitario Río Hortega de Valladolid, 47012 Valladolid, Spain; (11) Instituto de Investigación Biomédica de Salamanca (IBSAL), Gerencia Regional de Salud de Castilla y León, 47007 Valladolid, Spain; (12) Critical Care Department, Hospital Joan XXIII, 43005 Tarragona, Spain; (13) Anestesia, Reanimación y Terapia del Dolor, Hospital Universitario de Basurto, 48013 Bilbao, Spain; (14) Intensive Care Clinical Unit, Hospital Universitario Virgen de Rocío, 41013 Sevilla, Spain; (15) Hospital Santa Maria, IRBLleida, 25198 Lleida, Spain; (16) Critical Care Department, Hospital Universitario HM Montepríncipe, Universidad San Pablo-CEU, 28660 Madrid, Spain; (17) Servicio de Medicina Intensiva, Hospital Universitario Ramón y Cajal, 28034 Madrid, Spain; (18) Department of Intensive Care Medicine, Hospital Clínico Universitario Valladolid, 47003 Valladolid, Spain; (19) Critical Care Department, Hospital Universitari Arnau de Vilanova, IRBLleida, 25198 Lleida, Spain; (20) Hospital Álvaro Cunqueiro, 36213 Vigo, Spain; (21) Intensive Care Unit, Hospital Clínico y Universitario de Valencia, 46010 Valencia, Spain; (22) Department of Intensive Care Medicine, Hospital Universitario 12 de Octubre, 28041 Madrid, Spain, (23) Department of Medicine, CHUS, Complejo Hospitalario Universitario de Santiago, 15076 Santiago de Compostela, Spain; (24) Department of Intensive Care, Hospital Universitari de Bellvitge, L’Hospitalet de Llobregat, 08907 Barcelona, Spain; (25) Hospital de Mataró de Barcelona, 08301 Mataró, Spain; (26) Department of Medicine, Universitat Autònoma de Barcelona (UAB), Critical Care Department, Corpo-Ració Sanitària Parc Taulí, Sabadell, 08208 Barcelona, Spain; (27) Unidad de Cuidados Intensivos, Hospital San Pedro de Alcántara, 10003 Cáceres, Spain; (28) Intensive Care Unit, Hospital San Juan de Dios del Aljarafe, 41930 Sevilla, Spain; (29) Intensive Care Clinical Unit, Hospital Universitario Virgen Macarena, 41009 Seville, Spain; (30) Hospital General Universitario Gregorio Marañón, 28009 Madrid, Spain; (31) Intensive Care Department, Hospital Nuestra Señora de Gracia, 50009 Zaragoza, Spain; (32) Unidad de Medicina Intensiva, Hospital Universitario Virgen de Valme, 41014 Sevilla, Spain; (33) Critical Care Department, Hospital del Mar-IMIM, 08003 Barcelona, Spain; (34) Department of Intensive Medicine, Hospital Universitario Infanta Leonor, 28031 Madrid, Spain; (35) Critical Care Department, Hospital Universitario Lucus Augusti, 27003 Lugo, Spain; (36) Critical Care Department, Complejo Asistencial Universitario de Palencia, 34005 Palencia, Spain; (37) Departamento de Biología Funcional, Instituto Universitario de Oncología del Principado de Asturias, Universidad de Oviedo, 33011 Oviedo, Spain; (38) Instituto de Investigación Sanitaria del Principado de Asturias, Hospital Central de Asturias, 33011 Oviedo, Spain; (39) Servei de Medicina Intensiva, Hospital Universitari Son Espases, Palma de Mallorca, 07120 Illes Balears, Spain; (40) Servicio de Medicina Intensiva, Hospital Universitario Marqués de Valdecilla, 39008 Santander, Spain; (41) Servei de Medicina Intensiva, Hospital Universitari Germans Trias, 08916 Badalona, Spain; (42) Hospital Verge de la Cinta, 08916 Tortosa, Spain; (43) Hospital de Sant Joan d’Alacant, 03550 Alacant, Spain; (44) Servicio de Medicina Intensiva, Hospital Universitario y Politécnico La Fe, 46026 Valencia, Spain; (45) Intensive Care Unit, Hospital Son Llàtzer, Palma de Mallorca, 07198 Illes Balears, Spain; (46) Critical Care Department, Hospital Dr. Negrín., 35019 Las Palmas de GC, Spain; (47) Critical Care Department, Hospital Universitario Río Hortega de Valladolid, 47102 Valladolid, Spain; (48) Intensive Care Unit, Complexo Hospitalario Universitario de Ourense, 32005 Ourense, Spain.Postprint (published version

The nanomechanics of neurotoxina proteins reveals common features at the start of the neurodegeneration cascade.

1 pags. -- 56th Annual Meeting of the Biophysical-Society, FEB 25-29, 2012, San Diego, CAAmyloidogenic neurodegenerative diseases are incurable conditions caused by specific largely disordered proteins. However, the underlying molecular mechanism remains elusive. A favored hypothesis postulates that a critical conformational change in the monomer (an ideal therapeutic target) in these ‘‘neurotoxic proteins’’ triggers the pathogenic cascade. Using force spectroscopy with unequivocal singlemolecule identification we demonstrate a rich conformational polymorphism at their monomer level. This polymorphism strongly correlates with amyloidogenesis and neurotoxicity: it is absent in a fibrillization-incompetent mutant, favored by familial-disease mutations and diminished by a surprisingly promiscuous inhibitor of the monomeric b-conformational change and neurodegeneration. The demonstrated ability to inhibit the conformational heterogeneity of these proteins by a single pharmacological agent reveals common features in the monomer and suggests a common pathway to diagnose, prevent, halt or reverse multiple neurodegenerative disease

Risk factors associated with mortality among elderly patients with COVID-19: Data from 55 intensive care units in Spain

On behalf of CIBERESUCICOVID Project (COV20/00110ISCIII).Introduction and objectives: Critically-ill elderly ICU patients with COVID-19 have poor outcomes. We aimed to compare the rates of in-hospital mortality between non-elderly and elderly critically-ill COVID-19 ventilated patients, as well as to analyze the characteristics, secondary outcomes and independent risk factors associated with in-hospital mortality of elderly ventilated patients. Patients and Methods: We conducted a multicentre, observational cohort study including consecutive critically-ill patients admitted to 55 Spanish ICUs due to severe COVID-19 requiring mechanical ventilation (non-invasive respiratory support [NIRS; include non-invasive mechanical ventilation and high-flow nasal cannula] and invasive mechanical ventilation [IMV]) between February 2020 and October 2021. Results: Out of 5,090 critically-ill ventilated patients, 1,525 (27%) were aged =70 years (554 [36%] received NIRS and 971 [64%] received IMV. In the elderly group, median age was 74 years (interquartile range 72–77) and 68% were male. Overall in-hospital mortality was 31% (23% in patients <70 years and 50% in those =70 years; p<0.001). In-hospital mortality in the group =70 years significantly varied according to the modality of ventilation (40% in NIRS vs. 55% in IMV group; p<0.001). Factors independently associated with in-hospital mortality in elderly ventilated patients were age (sHR 1.07 [95%CI 1.05–1.10], p<0.001); previous admission within the last 30 days (sHR 1.40 [95%CI 1.04–1.89], p = 0.027); chronic heart disease (sHR 1.21 [95%CI 1.01–1.44], p = 0.041); chronic renal failure (sHR 1.43 [95%CI 1.12- 1.82], p = 0.005); platelet count (sHR 0.98 [95% CI 0.98–0.99], p<0.001); IMV at ICU admission (sHR 1.41 [95% CI 1.16- 1.73], p<0.001); and systemic steroids (sHR 0.61 [95%CI 0.48- 0.77], p<0.001). Conclusions: Amongst critically-ill COVID-19 ventilated patients, those aged =70 years presented significantly higher rates of in-hospital mortality than younger patients. Increasing age, previous admission within the last 30 days, chronic heart disease, chronic renal failure, platelet count, IMV at ICU admission and systemic steroids (protective) all comprised independent factors for in-hospital mortality in elderly patientsThis study was supported by the Instituto de Salud Carlos III de Madrid (COV20/00110, ISCIII); Fondo Europeo de Desarrollo Regional (FEDER); "Una manera de hacer Europa"; and Centro de Investigacion Biomédica En Red - Enfermedades Respiratorias (CIBERES). DdGC has received financial support from the Instituto de Salud Carlos III (Miguel Servet 2020:CP20/00041), co-funded by European Social Fund (ESF)/ “Investing in your future”. CC received a grant from the Fondo de Investigacion Sanitaria ( PI19/00207), Instituto de Salud Carlos III, co-funded by the European Union.Peer ReviewedCIBERESUCICOVID Project Investigators: Víctor D. Gumucio- Sanguino, Rafael Manez: Hospital Universitario de Bellvitge, Barcelona. Jordi Sole-Violan, Felipe Rodríguez de Castro: Hospital Dr. Negrín, Las Palmas. Fernando SuarezSipmann: Hospital Universitario La Princesa, Madrid. Ruth Noemí Jorge García, María Mora Aznar: Hospital Nuestra Senora de Gracia, Zaragoza. Mateu Torres, María Martinez, Cynthia Alegre, Jordi Riera, Sofía Contreras: Hospital Universitari Vall d’Hebron, Barcelona. Jesus Caballero, Javier Trujillano, Montse Vallverdu, Miguel Leon, Mariona Badía, Begona Balsera, Lluís Servia, Judit Vilanova, Silvia Rodríguez, Neus Montserrat, Silvia Iglesias, Javier Prados, Sula Carvalho, Mar Miralbes, Josman Monclou, Gabriel Jimenez, Jordi Codina, Estela Val, Pablo Pagliarani, Jorge Rubio, Dulce Morales, Andres Pujol, Angels Furro, Beatriz García, Gerard Torres, Javier Vengoechea, David de Gonzalo-Calvo, Jessica Gonzalez, Silvia Gomez: Hospital Universitari Arnau de Vilanova, Lleida. Jose M. Gomez: Hospital General Universitario Gregorio Marañon, Madrid. Nieves Franco: Hospital Universitario de Mostoles, Madrid. Jose Barberan: Hospital Universitario HM Montepríncipe. Guillermo M Albaiceta, Lorena Forcelledo Espina, Emilio García Prieto, Paula Martín Vicente, Cecilia del Busto Martínez: Hospital Universitario Central de Asturias, Oviedo. Pablo Vidal: Complexo Hospitalario Universitario de Ourense, Ourense. Jose Luis García Garmendia, María Aguilar Cabello, Carmen Eulalia Martínez Fernandez: Hospital San Juan de Dios del Aljarafe, Sevilla. Nieves Carbonell, María Luisa Blasco Cortes, Ainhoa Serrano Lazaro, Mar Juan Díaz: Hospital Clínic Universitari de Valencia, Valencia. Aaron Blandino Ortiz:Hospital Universitario Ramon y Cajal, Madrid. Rosario Menendez: Hospital La Fe de Valencia. Luis Jorge Valdivia: Hospital Universitario de Leon, Leon. María Victoria Boado: Hospital Universitario de Cruces, Barakaldo. Susana Sancho Chinesta: Hospital Universitario y Politecnico La Fe, Valencia. Maria del Carmen de la Torre: Hospital de Mataro. Ignacio Martínez Varela, María Teresa Bouza Vieiro, Ines Esmorís Arij on: Hospital Universitario Lucus Augusti, Lugo. David Campi Hermoso., Rafaela Nogueras Salinas., Teresa Farre Monjo., Ramon Nogue Bou., Gregorio Marco Naya., Carme Barbera, Nuria Ramon Coll: Hospital Universitari de Santa Maria, Lleida. Mercedes Catalan-Gonzalez, Juan Carlos Montejo-Gonzalez: Hospital Universitario 12 de Octubre, Madrid. Gloria Renedo SanchezGiron, Juan Bustamante-Munguira, Elena Bustamante-Munguira, Ramon Cicuendez Avila, Nuria Mamolar Herrera: Hospital Clínico Universitario, Valladolid. Raquel Almansa: Instituto de Investigacion Biomedica de Salamanca (IBSAL). Víctor Sagredo: Hospital Universitario de Salamanca, Salamanca. Jose Anon, Alexander Agrifoglio, Lucia Cachafeiro, Emilio Maseda: Hospital Universitario La Paz-Carlos III, Madrid. Lorenzo Socias, Mariana Andrea Novo, Albert Figueras, Maria Teresa Janer, Laura Soliva, Marta Ocon, Luisa Clar, J Ignacio Ayestaran: Hospital Universitario Son Espases, Palma de Mallorca. Yhivian Penasco, Sandra Campos Fernandez: Hospital Universitario Marques de Valdecilla, Santander. Mireia Serra-Fortuny, Eva Forcadell-Ferreres, Immaculada Salvador-Adell, Neus Bofill, Berta Adell-Serrano, Josep Pedregosa Díaz, Nuria Casacuberta-Barbera, Luis Urrelo-Cerron, Angels Piñol-Tena, Ferran Roche-Campo: Hospital Verge de la Cinta de Tortosa, Tortosa. Amalia Martínez de la Gandara, Pablo Ryan Murua, Covadonga Rodríguez Ruíz, Laura Carrion García, Juan I Lazo Alvarez: Hospital Universitario Infanta Leonor, Madrid. Jose Angel Lorente: Hospital Universitario de Getafe. Ana Loza-Vazquez, Desire Macias Guerrero: Hospital Universitario Virgen de Valme, Sevilla. Arturo Huerta, Daniel Tognetti: Clinica Sagrada Familia, Barcelona. Carlos García Redruello, David Mosquera Rodríguez, Eva María Menor Fernandez, Sabela Vara Adrio, Vanesa Gomez Casal, Marta Segura Pensado, María Digna Rivas Vilas, Amaia García Sagastume: Hospital de Vigo, Vigo. Raul de Pablo Sanchez, David Pesta na Laguna, Tommaso Bardi: Hospital Universitario Ramon y Cajal, Madrid. Rosario Amaya Villar, Carmen Gomez Gonzalez, Maria Luisa Gascon Castillo: Hospital Universitario Virgen del Rocio, Sevilla. Jose Garnacho-Montero, María Luisa Canton-Bulnes: Hospital Universitario Virgen Macarena, Sevilla. Judith Marin-Corral, Cristina Carbajales Perez: Hospital Alvaro Cunqueiro, Vigo. Joan Ramon Masclans, Ana Salazar Degracia, Judit Bigas, Rosana Munoz-Bermudez, Clara Vila-Vilardel, Francisco Parrilla, Irene Dot, Ana Zapatero, Yolanda Díaz, María Pilar Gracia, Purificacion Perez, Andrea Castellví, Cristina Climent: Hospital del Mar, Barcelona. Lidia Serra, Laura Barbena, Iosune Cano: Consorci Sanitari del Maresme, Barcelona. Pilar Ricart, Alba Herraiz, Pilar Marcos, Laura Rodríguez, Maria Teresa Sarinena, Ana Sanchez: Hospital Universitari Germans Trias i Pujol, Badalona. Alejandro Ubeda: Hospital Punta de Europa, Algeciras. María Cruz Martin Delgado: Hospital Universitario Torrejon-Universidad Francisco de Vitoria, Madrid. Elena Gallego, Juan Fernando Masa Jimenez: Hospital Universitario San Pedro de Alcantara, Caceres. Gemma Goma, Emi Díaz: Hospital Parc Taulí, Sabadell. Mercedes Ibarz, Diego De Mendoza: Hospital Universitari Sagrat Cor, Bacelona. Enric Barbeta, Victoria Alcaraz-Serrano, Joan Ramon Badia, Manuel Castella, Leticia Bueno, Adrian Ceccato, Andrea Palomeque, Laia Fernandez Barat, Catia Cilloniz, Pamela Conde, Javier Fernandez, Albert Gabarrus, Karsa Kiarostami, Alexandre Lopez- Gavín, Cecilia L Mantellini, Carla Speziale, Nil Vazquez, Hua Yang, Minlan Yang, Carlos Ferrando, Pedro Castro, Marta Arrieta, Jose Maria Nicolas, Rut Andrea: Hospital Clinic, Barcelona. Marta Barroso, Raquel Perez, Sergio Alvarez, Dario Garcia-Gasulla, Adrian Tormos: Barcelona supercomputing Center, Barcelona. Luis Tamayo Lomas, Cesar Aldecoa, Ruben Herran-Monge, Jose Angel Berezo García, Pedro Enríquez Giraudo: Hospital Rio Hortega, Valladolid. Pablo Cardinal Fernandez, Alberto Rubio Lopez, Orville Baez Pravia: Hospitales HM, Madrid. Juan Lopez Messa, Leire Perez Bastida, Antonjo Alvarez Ruiz: Complejo Asistencial Universitario de Palencia, Palencia. Jose Trenado, Anna Parera Pous: Hospital Universitari MutuaTerrassa, Terrassa. Cristobal Galban, Ana Lopez Lago, Eva Saborido Paz, Patricia Barral Segade: Hospital de Santiago de Compostela, Santiago. Ana Balan Marino, Manuel Valledor Mendez: Hospital San Agustin, Aviles. Raul de Frutos, Luciano Aguilera: Hospital Basurto, Basurto. Felipe Perez-García, Esther Lopez-Ramos, Angela Leonor Ruiz-García, Belen Betere: Hospital Universitario Principe Asturias, Alcala de Henares. Rafael Blancas: Hospital Universitario del Tajo, Aranjuez. Cristina Dolera, Gloria Perez Planelles, Enrique Marmol Peis, Maria Dolores Martinez Juan, Miriam Ruiz Miralles, Eva Perez Rubio, Maria Van der Hofstadt MartinMontalvo, Angel Sanchez-Miralles, Tatiana Villada Warrington: Hospital Universitario Sant Joan d’Alacant, Alicante. Juan Carlos Pozo-Laderas: Hospital Universitario Reina Sofia. Angel Estella, Sara Guadalupe Moreno Cano: Hospital de Jerez, Jerez. Federico Gordo: Hospital Universitario del Henares, Coslada. Basilisa Martinez Palacios: Hospital Universitario Infanta Cristina, Parla. Maite Nieto, Maria Teresa Nieto: Hospital de Segovia, Segovia. Sergio Ossa: Hospital de Burgos, Burgos. Ana Ortega: Hospital Montecelo, Pontevedra. Miguel Sanchez: Hospital Clinico, Madrid. Bitor Santacoloma: Hospital Galdakao, Galdakao.Postprint (published version

Matrix Metalloproteinase-9 Expression Is Associated with the Absence of Response to Neoadjuvant Chemotherapy in Triple-Negative Breast Cancer Patients

Triple-negative breast cancer (TNBC) is particularly challenging due to the weak or absent response to therapeutics and its poor prognosis. The effectiveness of neoadjuvant chemotherapy (NAC) response is strongly influenced by changes in elements of the tumor microenvironment (TME). This work aimed to characterize the residual TME composition in 96 TNBC patients using immunohistochemistry and in situ hybridization techniques and evaluate its prognostic implications for partial responders vs. non-responders. Compared with non-responders, partial responders containing higher levels of CD83+ mature dendritic cells, FOXP3+ regulatory T cells, and IL-15 expression but lower CD138+ cell concentration exhibited better OS and RFS. However, along with tumor diameter and positive nodal status at diagnosis, matrix metalloproteinase-9 (MMP-9) expression in the residual TME was identified as an independent factor associated with the impaired response to NAC. This study yields new insights into the key components of the residual tumor bed, such as MMP-9, which is strictly associated with the lack of a pathological response to NAC. This knowledge might help early identification of TNBC patients less likely to respond to NAC and allow the establishment of new therapeutic targets

Prognostic Implications of the Residual Tumor Microenvironment after Neoadjuvant Chemotherapy in Triple-Negative Breast Cancer Patients without Pathological Complete Response

Simple Summary Triple-negative breast cancer (TNBC) is currently in the clinical research spotlight because of the tumor's aggressive and invasive nature and the scarcity of therapeutic targets. Despite recent advances in identifying reliable prognostic biomarkers in the tumor microenvironment (TME), rigorous evaluation of their predictive capacity remains challenging. We describe the immune cellular and genetic profile of the residual tumor of TNBC that does not achieve a pathological complete response (pCR) after neoadjuvant chemotherapy (NAC). A high concentration of lymphocytes and dendritic cells, as well as genetic TME markers such as MUC-1 and CXCL13 in the residual tumor, are valuable prognostic factors of survival and relapse in TNBC patients. From a clinical health perspective, a thorough understanding of the composition of the TME and its prognostic implications might yield relevant immunological information and reveal key predictive biomarkers. This could ultimately help substantially improve the outcomes of residual cancer-burdened TNBC patients after NAC. With a high risk of relapse and death, and a poor or absent response to therapeutics, the triple-negative breast cancer (TNBC) subtype is particularly challenging, especially in patients who cannot achieve a pathological complete response (pCR) after neoadjuvant chemotherapy (NAC). Although the tumor microenvironment (TME) is known to influence disease progression and the effectiveness of therapeutics, its predictive and prognostic potential remains uncertain. This work aimed to define the residual TME profile after NAC of a retrospective cohort with 96 TNBC patients by immunohistochemical staining (cell markers) and chromogenic in situ hybridization (genetic markers). Kaplan-Meier curves were used to estimate the influence of the selected TME markers on five-year overall survival (OS) and relapse-free survival (RFS) probabilities. The risks of each variable being associated with relapse and death were determined through univariate and multivariate Cox analyses. We describe a unique tumor-infiltrating immune profile with high levels of lymphocytes (CD4, FOXP3) and dendritic cells (CD21, CD1a and CD83) that are valuable prognostic factors in post-NAC TNBC patients. Our study also demonstrates the value of considering not only cellular but also genetic TME markers such as MUC-1 and CXCL13 in routine clinical diagnosis to refine prognosis modelling