Técnicas avanzadas de medida de dispersión cromática a frecuencias ópticas

La fibra óptica a pesar de tener buenas propiedades, también presenta limitaciones, una de estas limitaciones es la dispersión. La dispersión es un fenómeno que introduce un retardo diferente a cada una de las componentes espectrales que se transmiten por la fibra óptica. Este proyecto esta basado en el estudio de este fenómeno y la medida del mismo a partir de métodos conocidos y propuestas de mejoras de estos. Se realiza un estudio matemático genérico del esquema básico de medida que tiene en cuenta todos los parámetros que pueden controlarse de forma externa, con el objetivo de identificar nuevos métodos de medida usando el mismo esquema experimental básico. Además, se monta en el laboratorio y se automatiza uno de los procesos básicos para la realización de las medidas, la determinación de la función de transferencia del modulador Mach-Zehnde

Pharmacological modulation of SAMHD1 activity by CDK4/6 inhibitors improves anticancer therapy

Funding: This research was funded by Instituto de Salud Carlos III, Fondo de Investigación Sanitaria (FIS) PI16/00103, PI17/00624 and CP14/00016 cofinanced by FEDER. EB is a research fellow from ISCIII-FIS (CP14/00016). EGV, MP, LG are research fellows from Generalitat de Catalunya AGAUR. RB is a research fellow from PERIS, Generalitat de Catalunya (PERIS SLT002/16/00059). IE is a research fellow from la Caixa Bank Foundation (LCF/BQ/IN18/11660017) cofunded by the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No. 713673.Sterile alpha motif and histidine-aspartic acid domain-containing protein 1 (SAMHD1) is a dNTP triphosphohydrolase involved in the regulation of the intracellular dNTP pool, linked to viral restriction, cancer development and autoimmune disorders. SAMHD1 function is regulated by phosphorylation through a mechanism controlled by cyclin-dependent kinases and tightly linked to cell cycle progression. Recently, SAMHD1 has been shown to decrease the efficacy of nucleotide analogs used as chemotherapeutic drugs. Here, we demonstrate that SAMHD1 can enhance or decrease the efficacy of various classes of anticancer drug, including nucleotide analogues, but also anti-folate drugs and CDK inhibitors. Importantly, we show that selective CDK4/6 inhibitors are pharmacological activators of SAMHD1 that act by inhibiting its inactivation by phosphorylation. Combinations of a CDK4/6 inhibitor with nucleoside or folate antimetabolites potently enhanced drug efficacy, resulting in highly synergic drug combinations (CI < 0.04). Mechanistic analyses reveal that cell cycle-controlled modulation of SAMHD1 function is the central process explaining changes in anticancer drug efficacy, therefore providing functional proof of the potential of CDK4/6 inhibitors as a new class of adjuvants to boost chemotherapeutic regimens. The evaluation of SAMHD1 expression in cancer tissues allowed for the identification of cancer types that would benefit from the pharmacological modulation of SAMHD1 function. In conclusion, these results indicate that the modulation of SAMHD1 function may represent a promising strategy for the improvement of current antimetabolite-based treatment

IRF7 expression correlates with HIV latency reversal upon specific blockade of immune activation

The persistence of latent HIV reservoirs allows for viral rebound upon antiretroviral therapy interruption, hindering effective HIV-1 cure. Emerging evidence suggests that modulation of innate immune stimulation could impact viral latency and contribute to the clearing of HIV reservoir. Here, the latency reactivation capacity of a subclass of selective JAK2 inhibitors was characterized as a potential novel therapeutic strategy for HIV-1 cure. Notably, JAK2 inhibitors reversed HIV-1 latency in non-clonal lymphoid and myeloid in vitro models of HIV-1 latency and also ex vivo in CD4+ T cells from ART+ PWH, albeit its function was not dependent on JAK2 expression. Immunophenotypic characterization and whole transcriptomic profiling supported reactivation data, showing common gene expression signatures between latency reactivating agents (LRA; JAK2i fedratinib and PMA) in contrast to other JAK inhibitors, but with significantly fewer affected gene sets in the pathway analysis. In depth evaluation of differentially expressed genes, identified a significant upregulation of IRF7 expression despite the blockade of the JAK-STAT pathway and downregulation of proinflammatory cytokines and chemokines. Moreover, IRF7 expression levels positively correlated with HIV latency reactivation capacity of JAK2 inhibitors and also other common LRAs. Collectively, these results represent a promising step towards HIV eradication by demonstrating the potential of innate immune modulation for reducing the viral reservoir through a novel pathway driven by IRF7

Plasma proteomic profiling identifies CD33 as a marker of HIV control in natural infection and after therapeutic vaccination

Altres ajuts: National Institutes of Health (NIH), P01-AI131568Biomarkers predicting the outcome of HIV-1 virus control in natural infection and after therapeutic interventions in HIV-1 cure trials remain poorly defined. The BCN02 trial (NCT02616874), combined a T-cell vaccine with romidepsin (RMD), a cancer-drug that was used to promote HIV-1 latency reversal and which has also been shown to have beneficial effects on neurofunction. We conducted longitudinal plasma proteomics analyses in trial participants to define biomarkers associated with virus control during monitored antiretroviral pause (MAP) and to identify novel therapeutic targets that can improve future cure strategies. BCN02 was a phase I, open-label, single-arm clinical trial in early-treated, HIV infected individuals. Longitudinal plasma proteomes were analyzed in 11 BCN02 participants, including 8 participants that showed a rapid HIV-1 plasma rebound during a monitored antiretroviral pause (MAP-NC, 'non-controllers') and 3 that remained off ART with sustained plasma viremia <2000 copies/ml (MAP-C, 'controllers'). Inflammatory and neurological proteomes in plasma were evaluated and integration data analysis (viral and neurocognitive parameters) was performed. Validation studies were conducted in a cohort of untreated HIV-1+ individuals (n = 96) and in vitro viral replication assays using an anti-CD33 antibody were used for functional validation

Heterogeneous Infectivity and Pathogenesis of SARS-CoV-2 Variants Beta, Delta and Omicron in Transgenic K18-hACE2 and Wildtype Mice

Altres ajuts: Fundació La Marató de TV3 202126-30-21The emerging SARS-CoV-2 variants of concern (VOCs) may display enhanced transmissibility, more severity and/or immune evasion; however, the pathogenesis of these new VOCs in experimental SARS-CoV-2 models or the potential infection of other animal species is not completely understood. Here we infected K18-hACE2 transgenic mice with B.1, B.1.351/Beta, B.1.617.2/Delta and BA.1.1/Omicron isolates and demonstrated heterogeneous infectivity and pathogenesis. B.1.351/Beta variant was the most pathogenic, while BA.1.1/Omicron led to lower viral RNA in the absence of major visible clinical signs. In parallel, we infected wildtype (WT) mice and confirmed that, contrary to B.1 and B.1.617.2/Delta, B.1.351/Beta and BA.1.1/Omicron can infect them. Infection in WT mice coursed without major clinical signs and viral RNA was transient and undetectable in the lungs by day 7 post-infection. In silico modeling supported these findings by predicting B.1.351/Beta receptor binding domain (RBD) mutations result in an increased affinity for both human and murine ACE2 receptors, while BA.1/Omicron RBD mutations only show increased affinity for murine ACE2

Complex multiple risk intervention to promote healthy behaviours in people between 45 to 75 years attended in primary health care (EIRA study): study protocol for a hybrid trial

CDATA[CDATA[Background: Health promotion is a key process of current health systems Primary Health Care (PHC) is the ideal setting for health promotion but multifaceted barriers make its integration difficult in the usual care. The majority of the adult population engages two 01 more risk behaviours, that is why a multiple intervention might be more effective and efficient The primary objectives are to evaluate the effectiveness, the cost effectiveness and an implementation strategy of a complex multiple risk intervention to promote healthy behaviours in people between 45 to 75 years attended in PHC. CDATA[CDATA[Methods: This study is a cluster randomised controlled hybrid type 2 trial with two parallel groups comparing a complex multiple risk behaviour intervention with usual care It will be carried out in 26 PHC centres in Spam The study focuses on people between 45 and 75 years who carry out two or more of the following unhealthy behaviours tobacco use, low adherence to the Mediterranean dietary pattern or insufficient physical activity level The intervention is based on the Transtheoretical Model and it will be made by physicians and nurses in the routine care of PHC practices according to the conceptual framework of the ''5A''s" It will have a maximum duration of 12 months and it will be carried out to three different levels (individual, group and community) Incremental cost per quality adjusted life year gamed measured by the tanffs of the EuioQo! 5D questionnaire will be estimated. The implementation strategy is based on the ''Consolidated Framework for Implementation Research, a set of discrete implementation strategies and an evaluation framework. CDATA[CDATA[Discussion: EIRA study will determine the effectiveness and cost effectiveness of a complex multiple risk intervention and will provide a better understanding of implementation processes of health promotion interventions in PHC setting. It may contribute to increase knowledge about the individual and structural barriers that affect implementation of these interventions and to quantify the contextual factors that moderate the effectiveness of implementation

Identification and characterization of novel latency-reversing agents to clear HIV-1 viral reservoir

La terapia antirretroviral ha cambiado la perspectiva sobre la infección por VIH-1 de enfermedad letal a crónica. Aun así, el reservorio latente del VIH-1 es una de las mayores barreras para lograr una cura. La estrategia “shock and kill” se basa en inducir la transcripción viral del provirus latente del VIH-1 seguido de la muerte selectiva de las células reactivadas. Aunque muchos agentes reversores de la latencia (LRAs) han sido identificados y testados, ninguno ha logrado erradicar eficazmente dicho reservorio. Dada la necesidad de nuevos agentes y estrategias capaces de eliminar el reservorio latente, hemos propuesto moduladores de la respuesta inmune innata o del ciclo celular para dicho fin. El estudio de moduladores de la inmunidad innata puede representar una alternativa dadas sus funciones intrínsecas, i. e., protección y eliminación de infecciones. Acitretina, regulador de la inmunidad innata aprobado contra la psoriasis, ha sido propuesto como inductor de la reactivación del VIH-1 y la muerte de las células infectadas. A pesar de ello, el efecto de acitretina en la reactivación fue muy modesto en la mayoría de modelos celulares que testamos, aunque se detectó la activación de la vía de RIG-I, y una ligera inducción de la reactivación viral en un modelo no clonal de células T. Además, acitretina tampoco promovió la eliminación de las células infectadas. Los compuestos anti-cáncer también han sido propuestos como estrategia contra el reservorio, debido a la habilidad de ciertos agentes para modificar la transcripción génica o promover la apoptosis. El cribado de una librería de compuestos anti-cancer reportó varias dianas cuya inhibición reactivó el VIH-1, incluyendo histona deacetilasas (HDACs), Janus quinasas (JAKs), IκB quinasas (IKKs) y proteínas “heat-shock” (HSPs). Entre los nuevos LRAs identificados, los inhibidores de Aurora kinasas (AURKi) representaron la mayor familia de compuestos, no descritos como LRAs, que mostraron capacidad de reactivación del VIH-1 de forma significativa y consistente. Los AURKi mejoraron la reactivación mediada por los HDACi, sugiriendo la habilidad para reactivar distintos provirus insertados. Curiosamente, AURKi restringió la replicación aguda del VIH-1, insinuando un papel dual para dichos compuestos en la infección por VIH-1. Midostaurin, un inhibidor de multi-quinasas aprobado contra la leucemia, también se identificó como LRA. Midostaurin reactivó el VIH-1, tanto por si solo como en combinación con otros LRAs, corroborando previos reportes que asociaron esa actividad con la activación de la vía de NF-κB. Además, también se observó una inhibición de la infección aguda del VIH-1 en células primarias dependiente de SAMHD1 no descrita. El hecho de que los AURKi y midostaurina mejoren la reactivación del VIH-1 en combinación con otros LRAs, corrobora la idea de que distintos compuestos pueden ser necesarios para reactivar todos los provirus integrados, presentando así distintas especificidades para la reactivación del provirus que dependan de su lugar de integración en el genoma. Estas observaciones plantean dudas sobre los modelos usados para estudiar la latencia del VIH-1, pues los modelos clonales podrían ser inadecuados por la falta de heterogeneidad de lugares de integración. En conjunto, nuestros resultados sugieren que la modulación de la inmunidad innata y ciclo celular podrían incluirse en el desarrollo de futuros LRAs para la estrategia “shock and kill”; aun así, investigaciones adicionales siguen siendo necesarias con tal de avanzar hacia la cura del VIH-1.Current antiretroviral therapy has changed the perspective of HIV-1 infection from a lethal illness to a chronic disease. However, the HIV-1 latent reservoir is a major hurdle to achieve a cure for HIV-1. The “shock and kill” strategy is based on inducing viral transcription of latent HIV-1 provirus followed by the selective killing of reactivated cells. Although several latency-reversing agents (LRAs) have been identified and tested, none of them has been able to efficiently eradicate the HIV-1 latent reservoir. Based on the need of novel agents and strategies to efficiently clear the latent reservoir, we evaluated compounds developed as modulators of the innate immune response or designed to modulate the cell cycle progression as novel agents able to purge the viral reservoir. The study of innate immune modulators as agents able to clear the HIV-1 reservoir might represent an alternative due to its intrinsic functions, i. e., protection and clearance of infections. The innate immune regulator acitretin, an FDA-approved compound for psoriasis, has been proposed to induce HIV-1 reactivation and selective killing of the infected cells. However, the effect of acitretin on HIV-1 reactivation was negligible in the vast majority of models tested, albeit activation of RIG-I pathway was detected and a mild induction of viral reactivation was observed in a non-clonal T cell model. Moreover, acitretin treatment did not induce the selective killing of the infected cells. Anti-cancer compounds have also been proposed as candidate therapies targeting the latent reservoir, mainly due to the ability of certain agents to modify gene transcription or to promote cell apoptosis. The assessment of the HIV-1 reactivation potential of an anti-cancer compound library reported several molecular targets whose inhibition promoted HIV-1 latency reversal, including the histone deacetylases (HDAC), Janus kinases (JAK), IκB kinases (IKKs) and heat shock proteins (HSPs). Among the new identified LRAs, Aurora kinases inhibitors (AURKi) represented the largest family of compounds not previously described as LRA that significantly and consistently showed HIV-1 reactivation capacity. AURKi were able to enhance the HDACi-mediated reactivation, suggesting that AURKi are able to target a distinct set of integrated provirus than that reactivated by the well-described HDAC inhibitors. Interestingly, AURKi restricted acute HIV-1 infection, suggesting a dual role for these compounds on HIV-1 infection. Midostaurin, a multi-kinase inhibitor approved for leukemia treatment, was also identified as an LRA. Midostaurin induced HIV-1 latency reactivation, either alone or in combination with other LRAs, consistent with previous reports that associated this activity with the activation of the innate immune NF-κB pathway. Moreover, we also observed a non-yet-reported and SAMHD1-dependent inhibitory effect of HIV-1 replication in primary cells. The enhanced capacity to promote HIV-1 reactivation of AURKi and midostaurin in combination with other LRAs supports the idea that different agents are needed to reactivate all latent provirus, presenting different specificities towards HIV-1 provirus reactivation depending on its integration site in the host genome. Furthermore, these observations also raise concerns on the models used to study HIV-1 latency, as clonal models might not be suitable due to the lack of heterogeneity in proviral insertion site, characteristic of non-clonal models. Altogether, our results suggest that modulation of innate immunity and cell cycle may be taken into account for the design of future LRAs for the “shock and kill” strategy; however, further research is still necessary before it can lead to an HIV-1 cure

Identification and characterization of novel latency-reversing agents to clear HIV-1 viral reservoir

La terapia antirretroviral ha cambiado la perspectiva sobre la infección por VIH-1 de enfermedad letal a crónica. Aun así, el reservorio latente del VIH-1 es una de las mayores barreras para lograr una cura. La estrategia “shock and kill” se basa en inducir la transcripción viral del provirus latente del VIH-1 seguido de la muerte selectiva de las células reactivadas. Aunque muchos agentes reversores de la latencia (LRAs) han sido identificados y testados, ninguno ha logrado erradicar eficazmente dicho reservorio. Dada la necesidad de nuevos agentes y estrategias capaces de eliminar el reservorio latente, hemos propuesto moduladores de la respuesta inmune innata o del ciclo celular para dicho fin. El estudio de moduladores de la inmunidad innata puede representar una alternativa dadas sus funciones intrínsecas, i. e., protección y eliminación de infecciones. Acitretina, regulador de la inmunidad innata aprobado contra la psoriasis, ha sido propuesto como inductor de la reactivación del VIH-1 y la muerte de las células infectadas. A pesar de ello, el efecto de acitretina en la reactivación fue muy modesto en la mayoría de modelos celulares que testamos, aunque se detectó la activación de la vía de RIG-I, y una ligera inducción de la reactivación viral en un modelo no clonal de células T. Además, acitretina tampoco promovió la eliminación de las células infectadas. Los compuestos anti-cáncer también han sido propuestos como estrategia contra el reservorio, debido a la habilidad de ciertos agentes para modificar la transcripción génica o promover la apoptosis. El cribado de una librería de compuestos anti-cancer reportó varias dianas cuya inhibición reactivó el VIH-1, incluyendo histona deacetilasas (HDACs), Janus quinasas (JAKs), IκB quinasas (IKKs) y proteínas “heat-shock” (HSPs). Entre los nuevos LRAs identificados, los inhibidores de Aurora kinasas (AURKi) representaron la mayor familia de compuestos, no descritos como LRAs, que mostraron capacidad de reactivación del VIH-1 de forma significativa y consistente. Los AURKi mejoraron la reactivación mediada por los HDACi, sugiriendo la habilidad para reactivar distintos provirus insertados. Curiosamente, AURKi restringió la replicación aguda del VIH-1, insinuando un papel dual para dichos compuestos en la infección por VIH-1. Midostaurin, un inhibidor de multi-quinasas aprobado contra la leucemia, también se identificó como LRA. Midostaurin reactivó el VIH-1, tanto por si solo como en combinación con otros LRAs, corroborando previos reportes que asociaron esa actividad con la activación de la vía de NF-κB. Además, también se observó una inhibición de la infección aguda del VIH-1 en células primarias dependiente de SAMHD1 no descrita. El hecho de que los AURKi y midostaurina mejoren la reactivación del VIH-1 en combinación con otros LRAs, corrobora la idea de que distintos compuestos pueden ser necesarios para reactivar todos los provirus integrados, presentando así distintas especificidades para la reactivación del provirus que dependan de su lugar de integración en el genoma. Estas observaciones plantean dudas sobre los modelos usados para estudiar la latencia del VIH-1, pues los modelos clonales podrían ser inadecuados por la falta de heterogeneidad de lugares de integración. En conjunto, nuestros resultados sugieren que la modulación de la inmunidad innata y ciclo celular podrían incluirse en el desarrollo de futuros LRAs para la estrategia “shock and kill”; aun así, investigaciones adicionales siguen siendo necesarias con tal de avanzar hacia la cura del VIH-1.Current antiretroviral therapy has changed the perspective of HIV-1 infection from a lethal illness to a chronic disease. However, the HIV-1 latent reservoir is a major hurdle to achieve a cure for HIV-1. The “shock and kill” strategy is based on inducing viral transcription of latent HIV-1 provirus followed by the selective killing of reactivated cells. Although several latency-reversing agents (LRAs) have been identified and tested, none of them has been able to efficiently eradicate the HIV-1 latent reservoir. Based on the need of novel agents and strategies to efficiently clear the latent reservoir, we evaluated compounds developed as modulators of the innate immune response or designed to modulate the cell cycle progression as novel agents able to purge the viral reservoir. The study of innate immune modulators as agents able to clear the HIV-1 reservoir might represent an alternative due to its intrinsic functions, i. e., protection and clearance of infections. The innate immune regulator acitretin, an FDA-approved compound for psoriasis, has been proposed to induce HIV-1 reactivation and selective killing of the infected cells. However, the effect of acitretin on HIV-1 reactivation was negligible in the vast majority of models tested, albeit activation of RIG-I pathway was detected and a mild induction of viral reactivation was observed in a non-clonal T cell model. Moreover, acitretin treatment did not induce the selective killing of the infected cells. Anti-cancer compounds have also been proposed as candidate therapies targeting the latent reservoir, mainly due to the ability of certain agents to modify gene transcription or to promote cell apoptosis. The assessment of the HIV-1 reactivation potential of an anti-cancer compound library reported several molecular targets whose inhibition promoted HIV-1 latency reversal, including the histone deacetylases (HDAC), Janus kinases (JAK), IκB kinases (IKKs) and heat shock proteins (HSPs). Among the new identified LRAs, Aurora kinases inhibitors (AURKi) represented the largest family of compounds not previously described as LRA that significantly and consistently showed HIV-1 reactivation capacity. AURKi were able to enhance the HDACi-mediated reactivation, suggesting that AURKi are able to target a distinct set of integrated provirus than that reactivated by the well-described HDAC inhibitors. Interestingly, AURKi restricted acute HIV-1 infection, suggesting a dual role for these compounds on HIV-1 infection. Midostaurin, a multi-kinase inhibitor approved for leukemia treatment, was also identified as an LRA. Midostaurin induced HIV-1 latency reactivation, either alone or in combination with other LRAs, consistent with previous reports that associated this activity with the activation of the innate immune NF-κB pathway. Moreover, we also observed a non-yet-reported and SAMHD1-dependent inhibitory effect of HIV-1 replication in primary cells. The enhanced capacity to promote HIV-1 reactivation of AURKi and midostaurin in combination with other LRAs supports the idea that different agents are needed to reactivate all latent provirus, presenting different specificities towards HIV-1 provirus reactivation depending on its integration site in the host genome. Furthermore, these observations also raise concerns on the models used to study HIV-1 latency, as clonal models might not be suitable due to the lack of heterogeneity in proviral insertion site, characteristic of non-clonal models. Altogether, our results suggest that modulation of innate immunity and cell cycle may be taken into account for the design of future LRAs for the “shock and kill” strategy; however, further research is still necessary before it can lead to an HIV-1 cure

Técnicas avanzadas de medida de dispersión cromática a frecuencias ópticas

La fibra óptica a pesar de tener buenas propiedades, también presenta limitaciones, una de estas limitaciones es la dispersión. La dispersión es un fenómeno que introduce un retardo diferente a cada una de las componentes espectrales que se transmiten por la fibra óptica. Este proyecto esta basado en el estudio de este fenómeno y la medida del mismo a partir de métodos conocidos y propuestas de mejoras de estos. Se realiza un estudio matemático genérico del esquema básico de medida que tiene en cuenta todos los parámetros que pueden controlarse de forma externa, con el objetivo de identificar nuevos métodos de medida usando el mismo esquema experimental básico. Además, se monta en el laboratorio y se automatiza uno de los procesos básicos para la realización de las medidas, la determinación de la función de transferencia del modulador Mach-Zehnde

Identification and characterization of novel latency-reversing agents to clear HIV-1 viral reservoir

La terapia antirretroviral ha cambiado la perspectiva sobre la infección por VIH-1 de enfermedad letal a crónica. Aun así, el reservorio latente del VIH-1 es una de las mayores barreras para lograr una cura. La estrategia "shock and kill" se basa en inducir la transcripción viral del provirus latente del VIH-1 seguido de la muerte selectiva de las células reactivadas. Aunque muchos agentes reversores de la latencia (LRAs) han sido identificados y testados, ninguno ha logrado erradicar eficazmente dicho reservorio. Dada la necesidad de nuevos agentes y estrategias capaces de eliminar el reservorio latente, hemos propuesto moduladores de la respuesta inmune innata o del ciclo celular para dicho fin. El estudio de moduladores de la inmunidad innata puede representar una alternativa dadas sus funciones intrínsecas, i. e., protección y eliminación de infecciones. Acitretina, regulador de la inmunidad innata aprobado contra la psoriasis, ha sido propuesto como inductor de la reactivación del VIH-1 y la muerte de las células infectadas. A pesar de ello, el efecto de acitretina en la reactivación fue muy modesto en la mayoría de modelos celulares que testamos, aunque se detectó la activación de la vía de RIG-I, y una ligera inducción de la reactivación viral en un modelo no clonal de células T. Además, acitretina tampoco promovió la eliminación de las células infectadas. Los compuestos anti-cáncer también han sido propuestos como estrategia contra el reservorio, debido a la habilidad de ciertos agentes para modificar la transcripción génica o promover la apoptosis. El cribado de una librería de compuestos anti-cancer reportó varias dianas cuya inhibición reactivó el VIH-1, incluyendo histona deacetilasas (HDACs), Janus quinasas (JAKs), IκB quinasas (IKKs) y proteínas "heat-shock" (HSPs). Entre los nuevos LRAs identificados, los inhibidores de Aurora kinasas (AURKi) representaron la mayor familia de compuestos, no descritos como LRAs, que mostraron capacidad de reactivación del VIH-1 de forma significativa y consistente. Los AURKi mejoraron la reactivación mediada por los HDACi, sugiriendo la habilidad para reactivar distintos provirus insertados. Curiosamente, AURKi restringió la replicación aguda del VIH-1, insinuando un papel dual para dichos compuestos en la infección por VIH-1. Midostaurin, un inhibidor de multi-quinasas aprobado contra la leucemia, también se identificó como LRA. Midostaurin reactivó el VIH-1, tanto por si solo como en combinación con otros LRAs, corroborando previos reportes que asociaron esa actividad con la activación de la vía de NF-κB. Además, también se observó una inhibición de la infección aguda del VIH-1 en células primarias dependiente de SAMHD1 no descrita. El hecho de que los AURKi y midostaurina mejoren la reactivación del VIH-1 en combinación con otros LRAs, corrobora la idea de que distintos compuestos pueden ser necesarios para reactivar todos los provirus integrados, presentando así distintas especificidades para la reactivación del provirus que dependan de su lugar de integración en el genoma. Estas observaciones plantean dudas sobre los modelos usados para estudiar la latencia del VIH-1, pues los modelos clonales podrían ser inadecuados por la falta de heterogeneidad de lugares de integración. En conjunto, nuestros resultados sugieren que la modulación de la inmunidad innata y ciclo celular podrían incluirse en el desarrollo de futuros LRAs para la estrategia "shock and kill"; aun así, investigaciones adicionales siguen siendo necesarias con tal de avanzar hacia la cura del VIH-1.Current antiretroviral therapy has changed the perspective of HIV-1 infection from a lethal illness to a chronic disease. However, the HIV-1 latent reservoir is a major hurdle to achieve a cure for HIV-1. The "shock and kill" strategy is based on inducing viral transcription of latent HIV-1 provirus followed by the selective killing of reactivated cells. Although several latency-reversing agents (LRAs) have been identified and tested, none of them has been able to efficiently eradicate the HIV-1 latent reservoir. Based on the need of novel agents and strategies to efficiently clear the latent reservoir, we evaluated compounds developed as modulators of the innate immune response or designed to modulate the cell cycle progression as novel agents able to purge the viral reservoir. The study of innate immune modulators as agents able to clear the HIV-1 reservoir might represent an alternative due to its intrinsic functions, i. e., protection and clearance of infections. The innate immune regulator acitretin, an FDA-approved compound for psoriasis, has been proposed to induce HIV-1 reactivation and selective killing of the infected cells. However, the effect of acitretin on HIV-1 reactivation was negligible in the vast majority of models tested, albeit activation of RIG-I pathway was detected and a mild induction of viral reactivation was observed in a non-clonal T cell model. Moreover, acitretin treatment did not induce the selective killing of the infected cells. Anti-cancer compounds have also been proposed as candidate therapies targeting the latent reservoir, mainly due to the ability of certain agents to modify gene transcription or to promote cell apoptosis. The assessment of the HIV-1 reactivation potential of an anti-cancer compound library reported several molecular targets whose inhibition promoted HIV-1 latency reversal, including the histone deacetylases (HDAC), Janus kinases (JAK), IκB kinases (IKKs) and heat shock proteins (HSPs). Among the new identified LRAs, Aurora kinases inhibitors (AURKi) represented the largest family of compounds not previously described as LRA that significantly and consistently showed HIV-1 reactivation capacity. AURKi were able to enhance the HDACi-mediated reactivation, suggesting that AURKi are able to target a distinct set of integrated provirus than that reactivated by the well-described HDAC inhibitors. Interestingly, AURKi restricted acute HIV-1 infection, suggesting a dual role for these compounds on HIV-1 infection. Midostaurin, a multi-kinase inhibitor approved for leukemia treatment, was also identified as an LRA. Midostaurin induced HIV-1 latency reactivation, either alone or in combination with other LRAs, consistent with previous reports that associated this activity with the activation of the innate immune NF-κB pathway. Moreover, we also observed a non-yet-reported and SAMHD1-dependent inhibitory effect of HIV-1 replication in primary cells. The enhanced capacity to promote HIV-1 reactivation of AURKi and midostaurin in combination with other LRAs supports the idea that different agents are needed to reactivate all latent provirus, presenting different specificities towards HIV-1 provirus reactivation depending on its integration site in the host genome. Furthermore, these observations also raise concerns on the models used to study HIV-1 latency, as clonal models might not be suitable due to the lack of heterogeneity in proviral insertion site, characteristic of non-clonal models. Altogether, our results suggest that modulation of innate immunity and cell cycle may be taken into account for the design of future LRAs for the "shock and kill" strategy; however, further research is still necessary before it can lead to an HIV-1 cure