Lentiviral Vector-Mediated Correction of a Mouse Model of Leukocyte Adhesion Deficiency Type I

Leukocyte adhesion deficiency type I (LAD-I) is a primary immunodeficiency caused by mutations in the ITGB2 gene and is characterized by recurrent and life-threatening bacterial infections. These mutations lead to defective or absent expression of β2 integrins on the leukocyte surface, compromising adhesion and extravasation at sites of infection. Three different lentiviral vectors (LVs) conferring ubiquitous or preferential expression of CD18 in myeloid cells were constructed and tested in human and mouse LAD-I cells. All three hCD18-LVs restored CD18 and CD11a membrane expression in LAD-I patient-derived lymphoblastoid cells. Corrected cells recovered the ability to aggregate and bind to sICAM-1 after stimulation. All vectors induced stable hCD18 expression in hematopoietic cells from mice with a hypomorphic Itgb2 mutation (CD18(HYP)), both in vitro and in vivo after transplantation of corrected cells into primary and secondary CD18(HYP) recipients. hCD18(+) hematopoietic cells from transplanted CD18(HYP) mice also showed restoration of mCD11a surface co-expression. The analysis of in vivo neutrophil migration in CD18(HYP) mice subjected to two different inflammation models demonstrated that the LV-mediated gene therapy completely restored neutrophil extravasation in response to inflammatory stimuli. Finally, these vectors were able to correct the phenotype of human myeloid cells derived from CD34(+) progenitors defective in ITGB2 expression. These results support for the first time the use of hCD18-LVs for the treatment of LAD-I patients in clinical trials

Autophagy modulates endothelial junctions to restrain neutrophil diapedesis during inflammation

The migration of neutrophils from the blood circulation to sites of infection or injury is a key immune response and requires the breaching of endothelial cells (ECs) that line the inner aspect of blood vessels. Unregulated neutrophil transendothelial cell migration (TEM) is pathogenic, but the molecular basis of its physiological termination remains unknown. Here, we demonstrated that ECs of venules in inflamed tissues exhibited a robust autophagic response that was aligned temporally with the peak of neutrophil trafficking and was strictly localized to EC contacts. Genetic ablation of EC autophagy led to excessive neutrophil TEM and uncontrolled leukocyte migration in murine inflammatory models, while pharmacological induction of autophagy suppressed neutrophil infiltration into tissues. Mechanistically, autophagy regulated the remodeling of EC junctions and expression of key EC adhesion molecules, facilitating their intracellular trafficking and degradation. Collectively, we have identified autophagy as a modulator of EC leukocyte trafficking machinery aimed at terminating physiological inflammation

A novel genetic variant in PTGS1 affects N-glycosylation of cyclooxygenase-1 causing a dominant-negative effect on platelet function and bleeding diathesis.

During platelet activation, arachidonic acid (AA) is released from membrane phospholipids and metabolized to thromboxane A2 (TXA2) through the actions of cyclooxygenase-1 (COX-1) and TXA2 synthase. Note, TXA2 binds to the platelet TXA2 receptor, causing shape change, secretion and platelet aggregation.1 Also, COX-1 (599aa; 70 kDa) has cyclooxygenase and peroxidase activities and it is functionally active as a homodimer, with each COX-1 monomer consisting of four highly conserved domains: an N-terminal signal peptide, a dimerization domain, a membrane-binding domain (MBD) and a large C-terminal catalytic domain2 (Figure 1A). Irreversible COX-1 inhibition by aspirin is a widely established anti-platelet therapy in cardiovascular disease.Fundación Mutua Madrileña, Grant/Award Number: AP172142019; Fundación Séneca, Grant/Award Number: 19873/GERM/15; Gerencia Regional de Salud, Grant/Award Numbers: 1647/A/17, 2061A/19; Instituto de Salud Carlos III (ISCIII) & Feder, Grant/Award Numbers: CB15/00055, PI17/01966, PI18/00598, PI20/00926, PI17/01311; Junta de Castilla y León; British Heart Foundation, Grant/Award Number: PG/17/40/33028; Ayuda a Grupos de Trabajo en Patología Hemorrágica; Premio López Borrasca 2019; Sociedad Española de Trombosis y Hemostasia

Lentiviral Vector-Mediated Correction of a Mouse Model of Leukocyte Adhesion Deficiency Type I

Leukocyte adhesion deficiency type I (LAD-I) is a primary immunodeficiency caused by mutations in the ITGB2 gene and is characterized by recurrent and life-threatening bacterial infections. These mutations lead to defective or absent expression of β2 integrins on the leukocyte surface, compromising adhesion and extravasation at sites of infection. Three different lentiviral vectors (LVs) conferring ubiquitous or preferential expression of CD18 in myeloid cells were constructed and tested in human and mouse LAD-I cells. All three hCD18-LVs restored CD18 and CD11a membrane expression in LAD-I patient-derived lymphoblastoid cells. Corrected cells recovered the ability to aggregate and bind to sICAM-1 after stimulation. All vectors induced stable hCD18 expression in hematopoietic cells from mice with a hypomorphic Itgb2 mutation (CD18(HYP)), both in vitro and in vivo after transplantation of corrected cells into primary and secondary CD18(HYP) recipients. hCD18(+) hematopoietic cells from transplanted CD18(HYP) mice also showed restoration of mCD11a surface co-expression. The analysis of in vivo neutrophil migration in CD18(HYP) mice subjected to two different inflammation models demonstrated that the LV-mediated gene therapy completely restored neutrophil extravasation in response to inflammatory stimuli. Finally, these vectors were able to correct the phenotype of human myeloid cells derived from CD34(+) progenitors defective in ITGB2 expression. These results support for the first time the use of hCD18-LVs for the treatment of LAD-I patients in clinical trials

XV International Congress of Control Electronics and Telecommunications: "The role of technology in times of pandemic and post-pandemic: innovation and development for strategic social and productive sectors"

La anterior selección, motivados por la aseveración de Manuel Castells -hace casi 20 años ya- que la innovación y la difusión de la tecnología parecía ser la herramienta apropiada para el desarrollo en la era de la información. Este 2020, sin embargo, ante la situación disruptiva que aquejó y aqueja a la sociedad red como una estructura social emergente de la Era de la Información basada en redes de producción, energizadas por el poder y la experiencia; falló y debe reencontrar su rumbo. Es así que los problemas acuciantes, ahora, fueron: la atención sanitaria y la superación de la epidemia de Sars Cov 2; tomó forma la, hasta entonces, visión irrealista de Castells que … no podemos avanzar con nuestros modelos de desarrollo actual, destruyendo nuestro entorno y excluyendo a la mayor parte de la humanidad de los beneficios de la revolución tecnológica más extraordinaria de la historia, sin sufrir una devastadora reacción por parte de la sociedad y la naturaleza. Fue así que el Cuarto Mundo, específicamente, donde la suficiencia de recurso humano, de capital, trabajo, información y mercado -vinculados todos a través de la tecnología- supuso que atendería eficazmente a través de la población que podía por su capacidad hacer uso racional y profesional del conocimiento, las necesidades de la mayoritaria población vulnerable y vulnerada. Por lo anterior, poner en el centro a las personas, en entornos de tarea y trabajo globales hiperconectados combinando espacios físicos, corrientes de información con canales de conexión expeditos, y formando profesionales del conocimiento que asuman y afronten los retos derivados de la transformación digital de empresas, universidades, y organizaciones, pero en condiciones de equidad y sujetos de prosperidad, será el desafío en los escenarios presentes y futuros inmediatos.The previous selection, motivated by the assertion of Manuel Castells -almost 20 years ago- that innovation and diffusion of technology seemed to be the appropriate tool for development in the information age. This 2020, however, in the face of the disruptive situation that afflicted and continues to afflict the network society as an emerging social structure of the Information Age based on production networks, energized by power and experience; He failed and must find his way again. Thus, the pressing problems now were: health care and overcoming the Sars Cov 2 epidemic; Castells' until then unrealistic vision took shape that... we cannot advance with our current development models, destroying our environment and excluding the majority of humanity from the benefits of the most extraordinary technological revolution in history, without suffering a devastating reaction from society and nature. It was thus that the Fourth World, specifically, where the sufficiency of human resources, capital, work, information and market - all linked through technology - meant that it would serve effectively through the population that could, due to its capacity, make rational use. and knowledge professional, the needs of the majority vulnerable and vulnerable population. Therefore, putting people at the center, in hyperconnected global task and work environments, combining physical spaces, information flows with expedited connection channels, and training knowledge professionals who assume and face the challenges derived from the digital transformation of companies, universities, and organizations, but in conditions of equality and subject to prosperity, will be the challenge in the present and immediate future scenarios.Bogot