Grupos Investigadores

Miembros del Grupo

Coordinador
Nguewa , Paul
(panguewa@unav.es)
Investigadores
Arola Verónica
Burguete Mikeo
Celia
Fernández Rubio
María Esther
Larrea Leoz
Manuel Francisco
Rubio Vallejo
Colaboradores
Miriam
Algarabel Olona
Lydia Grace
Evans
María
Fernández Gómez
José
Peña Guerrero
Eduardo
Sesma Herrero

Líneas de Investigación

  • Búsqueda y ensayos de tratamientos
  • COVID-19
  • Identificación y Estudio de Nuevas dianas terapéuticas
  • Infecciones emergentes
  • Inmunología de los patógenos
  • Mecanismos de virulencia de parásitos
  • Virus y Leishmania (Leishmania RNA Virus, LRV)

Palabras Clave

  • COVID-19
  • Diana terapéutica
  • Infecciones emergentes
  • Leishmania
  • Leishmaniasis
  • LRV (Leishmania RNA Virus)
  • Neglected Tropical Disease (NTD)
  • Patógeno
  • Respuesta inmune
  • Tratamiento
  • Virus

Publicaciones Científicas desde 2018

  • Autores: Rashidi, S.; Vieira, C.; Mansouri, R.; et al.
    Revista: PARASITOLOGY RESEARCH
    ISSN: 0932-0113 Vol.121 N° 7 2022 págs. 1853 - 1865
    Resumen
    Toxoplasma gondii is a pathogenic protozoan parasite belonging to the apicomplexan phylum that infects the nucleated cells of warm-blooded hosts leading to an infectious disease known as toxoplasmosis. Apicomplexan parasites such as T. gondii can display different mechanisms to control or manipulate host cells signaling at different levels altering the host subcellular genome and proteome. Indeed, Toxoplasma is able to modulate host cell responses (especially immune responses) during infection to its advantage through both structural and functional changes in the proteome of different infected cells. Consequently, parasites can transform the invaded cells into a suitable environment for its own replication and the induction of infection. Proteomics as an applicable tool can identify such critical proteins involved in pathogen (Toxoplasma)-host cell interactions and consequently clarify the cellular mechanisms that facilitate the entry of pathogens into host cells, and their replication and transmission, as well as the central mechanisms of host defense against pathogens. Accordingly, the current paper reviews several proteins (identified using proteomic approaches) differentially expressed in the proteome of Toxoplasma-infected host cells (macrophages and human foreskin fibroblasts) and tissues (brain and liver) and highlights their plausible functions in the cellular biology of the infected cells. The identification of such modulated proteins and their related cell impact (cell responses/signaling) can provide further information regarding parasite pathogenesis and biology that might lead to a better understanding of therapeutic strategies and novel drug targets.
  • Autores: Rashidi, S.; Fernández Rubio, Celia; Mansouri, R.; et al.
    Revista: PARASITOLOGY RESEARCH
    ISSN: 0932-0113 Vol.121 N° 1 2022 págs. 49 - 62
    Resumen
    The current drug treatments against protozoan parasitic diseases including Chagas, malaria, leishmaniasis, and toxoplasmosis represent good examples of drug resistance mechanisms and have shown diverse side effects. Therefore, the identification of novel therapeutic strategies and drug compounds against such life-threatening diseases is urgent. According to the successful usage of selenium (Se) compounds-based therapy against some diseases, this therapeutic strategy has been recently further underlined against these parasitic diseases by targeting different parasite's essential pathways. On the other hand, due to the important functions played by parasite selenoproteins in their biology (such as modulating the host immune response), they can be also considered as a novel therapeutic strategy by designing specific inhibitors against these important proteins. In addition, the immunomodulatory potentiality of these compounds to trigger T helper type 1 (Th1) cells and cytokine-mediated immune response for the substantial induction of proinflammatory cytokines, thus, Se, selenoproteins, and parasite selenoproteins could be further investigated to find possible vaccine antigens. Herein, we collect and present the results of some studies regarding Se-based therapy against protozoan parasitic diseases and highlight relevant information and some viewpoints that might be insightful to advance toward more effective studies in the future.
  • Autores: Rashidi, S.; Mansouri, R.; Ali-Hassanzadeh, M.; et al.
    Revista: ACTA TROPICA
    ISSN: 0001-706X Vol.231 2022 págs. 106431
    Resumen
    Micro RNAs (miRNAs), as regulators of gene expression at the post-transcriptional level, can respond to/or interact with cell signaling and affect the pathogenesis of different diseases/infections. The interaction/crosstalk of miRNAs with various cellular signaling networks including mTOR (as a master regulator of signaling relevant to different cellular mechanisms) might lead to the initiation, progression or restriction of certain disease processes. There are numerous studies that have identified the crosstalk between regulatory miRNA expression and the mTOR pathway (or mTOR signaling regulated by miRNAs) in different diseases which has a dual function in pathogenesis. However, the corresponding information in parasitic infections remains scarce. miRNAs have been suggested as specific targets for therapeutic strategies in several disorders such as parasitic infections. Thus, the targeting of miRNAs (as the modulators/regulators of mTOR) by small molecules and RNA-based therapeutics and consequently managing and modulating mTOR signaling and the downstream/related cell signaling/pathways might shed some light on the design of new therapeutic strategies against parasitic diseases, including Leishmaniasis. Accordingly, the present study attempts to highlight the importance of the crosstalk between regulatory miRNAs and mTOR signaling, and to review the relevant insights into parasitic infections by focusing specifically on Leishmania.
  • Autores: Rashidi, S.; Sánchez-Montejo, J.; Mansouri, R.; et al.
    Revista: ANIMALS
    ISSN: 2076-2615 Vol.12 N° 9 2022 págs. 1098
    Resumen
    Simple Summary The One Health concept to toxoplasmosis highlights that the health of humans is closely related to the health of animals and our common environment. Toxoplasmosis outcomes might be severe and fatal in patients with immunodeficiency, diabetes, and pregnant women and infants. Consequently, the development of effective vaccine and diagnostic strategies is urgent for the elimination of this disease. Proteomics analysis has allowed the identification of key proteins that can be utilized in the development of novel disease diagnostics and vaccines. This work presents relevant proteins found in the proteome of the life cycle-specific stages of Toxoplasma parasites. In fact, it brings together the main functionality key proteins from Toxoplasma parasites coming from proteomic approaches that are most likely to be useful in improving the disease management, and critically proposes innovative directions to finally develop promising vaccines and diagnostics tools. Toxoplasma gondii is a pathogenic protozoan parasite that infects the nucleated cells of warm-blooded hosts leading to an infectious zoonotic disease known as toxoplasmosis. The infection outcomes might be severe and fatal in patients with immunodeficiency, diabetes, and pregnant women and infants. The One Health approach to toxoplasmosis highlights that the health of humans is closely related to the health of animals and our common environment. The presence of drug resistance and side effects, the further improvement of sensitivity and specificity of serodiagnostic tools and the potentiality of vaccine candidates to induce the host immune response are considered as justifiable reasons for the identification of novel targets for the better management of toxoplasmosis. Thus, the identification of new critical proteins in the proteome of Toxoplasma parasites can also be helpful in designing and test more effective drugs, vaccines, and diagnostic tools. Accordingly, in this study we present important proteins found in the proteome of the life cycle-specific stages of Toxoplasma parasites that are potential diagnostic or vaccine candidates. The current study might help to understand the complexity of these parasites and provide a possible source of strategies and biomolecules that can be further evaluated in the pathobiology of Toxoplasma parasites and for diagnostics and vaccine trials against this disease.
  • Autores: Abdel Sater, F.; Younes, M.; Nassar, H.; et al.
    Revista: MOLECULAR BIOLOGY REPORTS
    ISSN: 0301-4851 Vol.49 N° 4 2022 págs. 3365
  • Autores: Rashidi, S.; Vieira, C.; Tuteja, R.; et al.
    Revista: BIOMOLECULES
    ISSN: 2218-273X Vol.12 N° 2 2022 págs. 257
    Resumen
    Human Leukocyte Antigen-G (HLA-G), a polymorphic non-classical HLA (HLA-Ib) with immune-regulatory properties in cancers and infectious diseases, presents both membrane-bound and soluble (sHLA-G) isoforms. Polymorphism has implications in host responses to pathogen infections and in pathogenesis. Differential expression patterns of HLA-G/sHLA-G or its polymorphism seem to be related to different pathological conditions, potentially acting as a disease progression biomarker. Pathogen antigens might be involved in the regulation of both membrane-bound and sHLA-G levels and impact immune responses during co-infections. The upregulation of HLA-G in viral and bacterial infections induce tolerance to infection. Recently, sHLA-G was found useful to identify the prognosis of Coronavirus disease 2019 (COVID-19) among patients and it was observed that the high levels of sHLA-G are associated with worse prognosis. The use of pathogens, such as Plasmodium falciparum, as immune modulators for other infections could be extended for the modulation of membrane-bound HLA-G in COVID-19-infected tissues. Overall, such information might open new avenues concerning the effect of some pathogens such as parasites in decreasing the expression level of HLA-G to restrict pathogenesis in some infections or to influence the immune responses after vaccination among others.
  • Autores: Larrea Leoz, María Esther; Fernández Rubio, Celia; Peña-Guerrero, J.; et al.
    Revista: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
    ISSN: 1422-0067 Vol.23 N° 21 2022 págs. 13203
    Resumen
    Around 15% of cancer cases are attributable to infectious agents. Epidemiological studies suggest that an association between leishmaniasis and cancer does exist. Recently, the homologue of PES1 in Leishmania major (LmjPES) was described to be involved in parasite infectivity. Mammalian PES1 protein has been implicated in cellular processes like cell cycle regulation. Its BRCT domain has been identified as a key factor in DNA damage-responsive checkpoints. This work aimed to elucidate the hypothetical oncogenic implication of BRCT domain from LmjPES in host cells. We generated a lentivirus carrying this BRCT domain sequence (lentiBRCT) and a lentivirus expressing the luciferase protein (lentiLuc), as control. Then, HEK293T and NIH/3T3 mammalian cells were infected with these lentiviruses. We observed that the expression of BRCT domain from LmjPES conferred to mammal cells in vitro a greater replication rate and higher survival. In in vivo experiments, we observed faster tumor growth in mice inoculated with lentiBRCT respect to lentiLuc HEK293T infected cells. Moreover, the lentiBRCT infected cells were less sensitive to the genotoxic drugs. Accordingly, gene expression profiling analysis revealed that BRCT domain from LmjPES protein altered the expression of proliferation- (DTX3L, CPA4, BHLHE41, BMP2, DHRS2, S100A1 and PARP9), survival- (BMP2 and CARD9) and chemoresistance-related genes (DPYD, Dok3, DTX3L, PARP9 and DHRS2). Altogether, our results reinforced the idea ...
  • Autores: El-Dirany, R.; Fernández Rubio, Celia; Peña Guerrero, José; et al.
    Revista: PHARMACEUTICS
    ISSN: 1999-4923 Vol.14 N° 11 2022 págs. 2528
    Resumen
    The lack of safe and cost-effective treatments against leishmaniasis highlights the urgent need to develop improved leishmanicidal agents. Antimicrobial peptides (AMPs) are an emerging category of therapeutics exerting a wide range of biological activities such as anti-bacterial, anti-fungal, anti-parasitic and anti-tumoral. In the present study, the approach of repurposing AMPs as antileishmanial drugs was applied. The leishmanicidal activity of two synthetic anti-lipopolysaccharide peptides (SALPs), so-called 19-2.5 and 19-4LF was characterized in Leishmania major. In vitro, both peptides were highly active against intracellular Leishmania major in mouse macrophages without exerting toxicity in host cells. Then, q-PCR-based gene profiling, revealed that this activity was related to the downregulation of several genes involved in drug resistance (yip1), virulence (gp63) and parasite proliferation (Cyclin 1 and Cyclin 6). Importantly, the treatment of BALB/c mice with any of the two AMPs caused a significant reduction in L. major infective burden. This effect was associated with an increase in Th1 cytokine levels (IL-12p35, TNF-alpha, and iNOS) in the skin lesion and spleen of the L. major infected mice while the Th2-associated genes were downregulated (IL-4 and IL-6). Lastly, we investigated the effect of both peptides in the gene expression profile of the P2X7 purinergic receptor, which has been reported as a therapeutic target in several diseases. The results showed significant repression of P2X7R by both peptides in the skin lesion of L. major infected mice to an extent comparable to that of a common anti-leishmanial drug, Paromomycin. Our in vitro and in vivo studies suggest that the synthetic AMPs 19-2.5 and 19-4LF are promising candidates for leishmaniasis treatment and present P2X7R as a potential therapeutic target in cutaneous leishmaniasis (CL).
  • Autores: Rashidi, S.; Mansouri, R.; Ali-Hassanzadeh, M.; et al.
    Revista: PARASITOLOGY
    ISSN: 0031-1820 Vol.148 N° 12 2021 págs. 1434 - 1446
    Resumen
    Leishmania parasites, the causative agents of leishmaniasis, are protozoan parasites with the ability to modify the signalling pathway and cell responses of their infected host cells. These parasite strategies alter the host cell environment and conditions favouring their replication, survival and pathogenesis. Since microRNAs (miRNAs) are able to post-transcriptionally regulate gene expression processes, these biomolecules can exert critical roles in controlling Leishmania-host cell interplay. Therefore, the identification of relevant miRNAs differentially expressed in Leishmania parasites as well as in infected cells, which affect the host fitness, could be critical to understand the infection biology, pathogenicity and immune response against these parasites. Accordingly, the current review aims to address the differentially expressed miRNAs in both, the parasite and infected host cells and how these biomolecules change cell signalling and host immune responses during infection. A deep understanding of these processes could provide novel guidelines and therapeutic strategies for managing and treating leishmaniasis.
  • Autores: Rashidi, S.; Fernández Rubio, Celia; Manzano-Roman, R.; et al.
    Revista: PARASITOLOGY
    ISSN: 0031-1820 Vol.148 N° 6 2021 págs. 655 - 671
    Resumen
    The association of leishmaniasis and malignancies in human and animal models has been highlighted in recent years. The misdiagnosis of coexistence of leishmaniasis and cancer and the use of common drugs in the treatment of such diseases prompt us to further survey the molecular biology of Leishmania parasites and cancer cells. The information regarding common expressed proteins, as possible therapeutic targets, in Leishmania parasites and cancer cells is scarce. Therefore, the current study reviews proteins, and investigates the regulation and functions of several key proteins in Leishmania parasites and cancer cells. The up- and down-regulations of such proteins were mostly related to survival, development, pathogenicity, metabolic pathways and vital signalling in Leishmania parasites and cancer cells. The presence of common expressed proteins in Leishmania parasites and cancer cells reveals valuable information regarding the possible shared mechanisms of pathogenicity and opportunities for therapeutic targeting in leishmaniasis and cancers in the future.
  • Autores: Rashidi, S.; Mansouri, R.; Ali-Hassanzadeh, M.; et al.
    Revista: PARASITOLOGY RESEARCH
    ISSN: 0932-0113 Vol.120 N° 4 2021 págs. 1151 - 1166
    Resumen
    The mechanistic (or mammalian) target of rapamycin (mTOR) is considered as a critical regulatory enzyme involved in essential signaling pathways affecting cell growth, cell proliferation, protein translation, regulation of cellular metabolism, and cytoskeletal structure. Also, mTOR signaling has crucial roles in cell homeostasis via processes such as autophagy. Autophagy prevents many pathogen infections and is involved on immunosurveillance and pathogenesis. Immune responses and autophagy are therefore key host responses and both are linked by complex mTOR regulatory mechanisms. In recent years, the mTOR pathway has been highlighted in different diseases such as diabetes, cancer, and infectious and parasitic diseases including leishmaniasis, toxoplasmosis, and malaria. The current review underlines the implications of mTOR signals and intricate networks on pathogen infections and the modulation of this master regulator by parasites. Parasitic infections are able to induce dynamic metabolic reprogramming leading to mTOR alterations in spite of many other ways impacting this regulatory network. Accordingly, the identification of parasite effects and interactions over such a complex modulation might reveal novel information regarding the biology of the abovementioned parasites and might allow the development of therapeutic strategies against parasitic diseases. In this sense, the effects of inhibiting the mTOR pathways are also considered in this context in the light of their potential for the prevention and treatment of parasitic diseases.
  • Autores: Rashidi, S.; Tuteja, R.; Mansouri, R.; et al.
    Revista: JOURNAL OF PROTEOMICS
    ISSN: 1874-3919 Vol.245 2021 págs. 104279
    Resumen
    There are important challenges when investigating individual post-translational modifications (PTMs) or protein interaction network and delineating if PTMs or their changes and cross-talks are involved during infection, disease initiation or as a result of disease progression. Proteomics and in silico approaches now offer the possibility to complement each other to further understand the regulatory involvement of these modifications in parasites and infection biology. Accordingly, the current review highlights key expressed or altered proteins and PTMs are invisible switches that turn on and off the function of most of the proteins. PTMs include phosphorylation, glycosylation, ubiquitylation, palmitoylation, myristoylation, prenylation, acetylation, methylation, and epigenetic PTMs in P. falciparum which have been recently identified. But also other low-abundant or overlooked PTMs that might be important for the parasite's survival, infectivity, antigenicity, immunomodulation and pathogenesis. We here emphasize the PTMs as regulatory pathways playing major roles in the biology, pathogenicity, metabolic pathways, survival, host-parasite interactions and the life cycle of P. falciparum. Further validations and functional characterizations of such proteins might confirm the discovery of therapeutic targets and might most likely provide valuable data for the treatment of P. falciparum, the main cause of severe malaria in human.
  • Autores: Kalantar, K.; Manzano-Roman, R.; Ghani, E.; et al.
    Revista: FUTURE MICROBIOLOGY
    ISSN: 1746-0913 Vol.16 N° 8 2021 págs. 607 - 613
    Resumen
    Apolipoprotein A-I (apo A-I) represents the main component of the Trypanosome lytic factor (TLF) which contributes to the host innate immunity against Trypanosoma and Leishmania. These parasites use complex and multiple strategies such as molecular mimicry to evade or subvert the host immune system. Previous studies have highlighted the adaptation mechanisms of TLF-resistant Trypanosoma species. These data might support the hypothesis that Leishmania parasites (amastigote forms in macrophages) might express apo A-I to bypass and escape from TLF action as a component of the host innate immune responses. The anti-inflammatory property of apo A-I is another mechanism that supports our idea that apo A-I may play a role in Leishmania parasites allowing them to bypass the host innate immune system.
  • Autores: El-Dirany, R.; Shahrour, Hawraa; Dirany Ahmad, Zeinab; et al.
    Revista: BIOMOLECULES
    ISSN: 2218-273X Vol.11 N° 7 2021 págs. 984
    Resumen
    Anti-microbial peptides (AMPs), small biologically active molecules, produced by different organisms through their innate immune system, have become a considerable subject of interest in the request of novel therapeutics. Most of these peptides are cationic-amphipathic, exhibiting two main mechanisms of action, direct lysis and by modulating the immunity. The most commonly reported activity of AMPs is their anti-bacterial effects, although other effects, such as anti-fungal, anti-viral, and anti-parasitic, as well as anti-tumor mechanisms of action have also been described. Their anti-parasitic effect against leishmaniasis has been studied. Leishmaniasis is a neglected tropical disease. Currently among parasitic diseases, it is the second most threating illness after malaria. Clinical treatments, mainly antimonial derivatives, are related to drug resistance and some undesirable effects. Therefore, the development of new therapeutic agents has become a priority, and AMPs constitute a promising alternative. In this work, we describe the principal families of AMPs (melittin, cecropin, cathelicidin, defensin, magainin, temporin, dermaseptin, eumenitin, and histatin) exhibiting a potential anti-leishmanial activity, as well as their effectiveness against other microorganisms.
  • Autores: Algarabel Olona, Miriam; Fernández Rubio, Celia; Musilova, K. ; et al.
    Revista: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
    ISSN: 1422-0067 Vol.22 N° 22 2021 págs. 12592
    Resumen
    Leishmaniasis is a neglected tropical disease caused by Leishmania spp. The improvement of existing treatments and the discovery of new drugs remain ones of the major goals in control and eradication of this disease. From the parasite genome, we have identified the homologue of the human oncogene PES1 in Leishmania major (LmjPES). It has been demonstrated that PES1 is involved in several processes such as ribosome biogenesis, cell proliferation and genetic transcription. Our phylogenetic studies showed that LmjPES encodes a highly conserved protein containing three main domains: PES N-terminus (shared with proteins involved in ribosomal biogenesis), BRCT (found in proteins related to DNA repair processes) and MAEBL-type domain (C-terminus, related to erythrocyte invasion in apicomplexan). This gene showed its highest expression level in metacyclic promastigotes, the infective forms; by fluorescence microscopy assay, we demonstrated the nuclear localization of LmjPES protein. After generating mutant parasites overexpressing LmjPES, we observed that these clones displayed a dramatic increase in the ratio of cell infection within macrophages. Furthermore, BALB/c mice infected with these transgenic parasites exhibited higher footpad inflammation compared to those inoculated with non-overexpressing parasites.
  • Autores: Sater, F. A.; Younes, M.; Nassar, H.; et al.
    Revista: MOLECULAR BIOLOGY REPORTS
    ISSN: 0301-4851 Vol.48 N° 11 2021 págs. 7243 - 7249
    Resumen
    Background The new SARS-CoV-2 variant VOC (202012/01), identified recently in the United Kingdom (UK), exhibits a higher transmissibility rate compared to other variants, and a reproductive number 0.4 higher. In the UK, scientists were able to identify the increase of this new variant through the rise of false negative results for the spike (S) target using a three-target RT-PCR assay (TaqPath kit). Methods To control and study the current coronavirus pandemic, it is important to develop a rapid and low-cost molecular test to identify the aforementioned variant. In this work, we designed primer sets specific to the VOC (202012/01) to be used by SYBR Green-based RT-PCR. These primers were specifically designed to confirm the deletion mutations Delta 69/Delta 70 in the spike and the Delta 106/Delta 107/Delta 108 in the NSP6 gene. We studied 20 samples from positive patients, detected by using the Applied Biosystems TaqPath RT-PCR COVID-19 kit (Thermo Fisher Scientific, Waltham, USA) that included the ORF1ab, S, and N gene targets. 16 samples displayed an S-negative profile (negative for S target and positive for N and ORF1ab targets) and four samples with S, N and ORF1ab positive profile. Results Our results emphasized that all S-negative samples harbored the mutations Delta 69/Delta 70 and Delta 106/Delta 107/Delta 108. This protocol could be used as a second test to confirm the diagnosis in patients who were already positive to COVID-19 but showed false negative results for S-gene. Conclusions This technique may allow to identify patients carrying the VOC (202012/01) or a closely related variant, in case of shortage in sequencing.
  • Autores: Penalvo, J. L. (Autor de correspondencia); Mertens, E.; Ademovic, E.; et al.
    Revista: BMJ OPEN
    ISSN: 2044-6055 Vol.11 N° 11 2021 págs. e055630
    Resumen
    Introduction unCoVer-Unravelling data for rapid evidence-based response to COVID-19-is a Horizon 2020-funded network of 29 partners from 18 countries capable of collecting and using real-world data (RWD) derived from the response and provision of care to patients with COVID-19 by health systems across Europe and elsewhere. unCoVer aims to exploit the full potential of this information to rapidly address clinical and epidemiological research questions arising from the evolving pandemic. Methods and analysis From the onset of the COVID-19 pandemic, partners are gathering RWD from electronic health records currently including information from over 22 000 hospitalised patients with COVID-19, and national surveillance and screening data, and registries with over 1 900 000 COVID-19 cases across Europe, with continuous updates. These heterogeneous datasets will be described, harmonised and integrated into a multi-user data repository operated through Opal-DataSHIELD, an interoperable open-source server application. Federated data analyses, without sharing or disclosing any individual-level data, will be performed with the objective to reveal patients' baseline characteristics, biomarkers, determinants of COVID-19 prognosis, safety and effectiveness of treatments, and potential strategies against COVID-19, as well as epidemiological patterns. These analyses will complement evidence from efficacy/safety clinical trials, where vulnerable, more complex/heterogeneous populations and those most at risk of severe COVID-19 are often excluded. Ethics and dissemination After strict ethical considerations, databases will be available through a federated data analysis platform that allows processing of available COVID-19 RWD without disclosing identification information to analysts and limiting output to data aggregates. Dissemination of unCoVer's activities will be related to the access and use of dissimilar RWD, as well as the results generated by the pooled analyses. Dissemination will include training and educational activities, scientific publications and conference communications.
  • Autores: Peña Guerrero, José; Fernández Rubio, Celia; Burguete-Mikeo, A.; et al.
    Revista: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
    ISSN: 1422-0067 Vol.22 N° 19 2021 págs. 10493
    Resumen
    Since many of the currently available antileishmanial treatments exhibit toxicity, low effectiveness, and resistance, search and validation of new therapeutic targets allowing the development of innovative drugs have become a worldwide priority. This work presents a structure-based drug discovery strategy to validate the Lmj_04_BRCT domain as a novel therapeutic target in Leishmania spp. The structure of this domain was explored using homology modeling, virtual screening, and molecular dynamics studies. Candidate compounds were validated in vitro using promastigotes of Leishmania major, L. amazonensis, and L. infantum, as well as primary mouse macrophages infected with L. major. The novel inhibitor CPE2 emerged as the most active of a group of compounds against Leishmania, being able to significantly reduce the viability of promastigotes. CPE2 was also active against the intracellular forms of the parasites and significantly reduced parasite burden in murine macrophages without exhibiting toxicity in host cells. Furthermore, L. major promastigotes treated with CPE2 showed significant lower expression levels of several genes (alpha-tubulin, Cyclin CYCA, and Yip1) related to proliferation and treatment resistance. Our in silico and in vitro studies suggest that the Lmj_04_BRCT domain and its here disclosed inhibitors are new potential therapeutic options against leishmaniasis.</p>
  • Autores: Mansouri, R.; Ali-Hassanzadeh, M.; Shafiei, R.; et al.
    Revista: PARASITOLOGY
    ISSN: 0031-1820 Vol.147 N° 12 2020 págs. 1255 - 1262
    Resumen
    Plasmodium falciparumis the main cause of severe malaria in humans that can lead to death. There is growing evidence of drug-resistance inP. falciparumtreatment, and the design of effective vaccines remains an ongoing strategy to control the disease. On the other hand, the recognition of specific diagnostic markers forP. falciparumcan accelerate the diagnosis of this parasite in the early stages of infection. Therefore, the identification of novel antigenic proteins especially by proteomic tools is urgent for vaccination and diagnosis ofP. falciparum. The proteome diversity of the life cycle stages ofP. falciparum, the altered proteome ofP. falciparum-infected human sera and altered proteins inP. falciparum-infected erythrocytes could be proposed as appropriate proteins for the aforementioned aims. Accordingly, this review highlights and proposes different proteins identified using proteomic approaches as promising markers in the diagnosis and vaccination ofP. falciparum.It seems that most of the candidates identified in this study were able to elicit immune responses in theP. falciparum-infected hosts and they also played major roles in the life cycle, pathogenicity and key pathways of this parasite.
  • Autores: Vacas Oleas, Andrés Fernando; Fernández Rubio, Celia; Larrea Leoz, María Esther; et al.
    Revista: BIOMEDICINES
    ISSN: 2227-9059 Vol.8 N° 11 2020 págs. 452
    Resumen
    A novel serine/threonine protein kinase, LmjF.22.0810, was recently described in Leishmania major. After generating an L. major cell line overexpressing LmjF.22.0810 (named LmJ3OE), the ability of this novel protein to modulate the Th2-type immune response was analyzed. Our results suggest that the protein kinase LmjF.22.0810 might be involved in leishmaniasis outcomes. Indeed, our study outlined the LmJ3OE parasites infectivity in vitro and in vivo. Transgenic parasites displayed lower phagocytosis rates in vitro, and their promastigote forms exhibited lower expression levels of virulence factors compared to their counterparts in control parasites. In addition, LmJ3OE parasites developed significantly smaller footpad swelling in susceptible BALB/c mice. Hematoxylin-eosin staining allowed the observation of a lower inflammatory infiltrate in the footpad from LmJ3OE-infected mice compared to animals inoculated with control parasites. Gene expression of Th2-associated cytokines and effectors revealed a dramatically lower induction in interleukin (IL)-4, IL-10, and arginase 1 (ARG1) mRNA levels at the beginning of the swelling; no expression change was found in Th1-associated cytokines except for IL-12. Accordingly, such results were validated by immunohistochemistry studies, illustrating a weaker expression of ARG1 and a similar induction for inducible NO synthase (iNOS) in footpads from LmJ3OE-infected mice compared to control L. major infected animals. Furthermore, the parasite burden was lower in footpads from LmJ3OE-infected mice. Our analysis indicated that such significant smaller footpad swellings might be due to an impairment of the Th2 immune response that subsequently benefits Th1 prevalence. Altogether, these studies depict LmjF.22.0810 as a potential modulator of host immune responses to Leishmania. Finally, this promising target might be involved in the modulation of infection outcome.
  • Autores: Rashidi, S.; Kalantar, K.; Fernández Rubio, Celia; et al.
    Revista: PATHOGENS AND DISEASE
    ISSN: 2049-632X Vol.78 N° 1 2020 págs. ftaa007
    Resumen
    Leishmaniasis includes a broad spectrum of pathological outcomes in humans caused by protozoan parasites from the genus Leishmania. In recent years, proteomic techniques have introduced novel proteins with critical functions in Leishmania parasites. Based on our report of a Chitin binding protein (CBP) in our previous immunoproteomic study, this article suggests that CBP might be an RNA binding protein (RBP) in Leishmania parasites. RBPs, as key regulatory factors, have a role in post-transcriptional gene regulation. The presence of RBPs in Leishmania parasites has not been considered so far; however, this study aims to open a new venue regarding RBPs in Leishmania parasites. Confirming CBP as an RBP in Leishmania parasites, exploring other RBPs and their functions might lead to interesting issues in leishmaniasis. In fact, due to the regulatory role of RBPs in different diseases including cancers and their further classification as therapeutic targets, the emerging evaluation of CBP and RBPs from Leishmania parasites may allow the discovery of novel and effective drugs against leishmaniasis.
  • Autores: RodrIguez-Losada, N.; de la Rosa Fernández-Pacheco, Francisco Javier; Larriva Hormigos, María; et al.
    Revista: JOURNAL OF ADVANCED RESEARCH
    ISSN: 2090-1232 Vol.23 2020 págs. 37 - 45
    Resumen
    Alpha-Synuclein (aSyn) is a chameleon-like protein. Its overexpression and intracellular deposition defines neurodegenerative alpha-synucleinopathies including Parkinson's disease. Whether aSyn upregulation is the cause or the protective reaction to alpha-synucleinopathies remains unresolved. Remarkably, the accumulation of aSyn is involved in cancer. Here, the neuroblastoma SH-SY5Y cell line was genetically engineered to overexpress aSyn at low and at high levels. aSyn cytotoxicity was assessed by the MIT and vital-dye exclusion methods, observed at the beginning of the sub-culture of low-aSyn overexpressing neurons when cells can barely proliferate exponentially. Conversely, high-aSyn overexpressing cultures grew at high rates while showing enhanced colony formation compared to low-aSyn neurons. Cytotoxicity of aSyn overexpression was indirectly revealed by the addition of pro-oxidant rotenone. Pretreatment with partially reduced graphene oxide, an apoptotic agent, increased toxicity of rotenone in low-aSyn neurons, but, it did not in high-aSyn neurons. Consistent with their enhanced proliferation, high-aSyn neurons showed elevated levels of SMP30, a senescence-marker protein, and the mitosis Ki-67 marker.
  • Autores: Pires, V. C. ; Magalhaes, C. P. ; Ferrante, M.; et al.
    Revista: ACTA TROPICA
    ISSN: 0001-706X Vol.211 2020
    Resumen
    17-N-allylamino-17-demethoxygeldanamycin (17-AAG, tanespimycin) is an inhibitor of heat shock protein 90 (Hsp90), which has been studied in the treatment of cancer such as leukemia or solid tumors. Alternatively, 17-AAG may represent a promising therapeutic agent against leishmaniasis. However, the delivery of 17-AAG is difficult due to its poor aqueous solubility. For exploring the therapeutic value of 17-AAG, we developed solid lipid nanoparticles (SLN) by double emulsion method. SLN exhibited similar to 100 nm, PDI < 0.2 and zeta potential similar to 20 mV. In addition, SLN were morphologically spherical with negligible aggregation. The entrapment efficiency of 17-AAG into the lipid matrix reached at nearly 80%. In a separate set of experiments, fluorescent SLN (FITC-labeled) showed a remarkable macrophage uptake, peaking within 2 h of incubation by confocal microscopy. Regarding the drug internalization as critical step for elimination of intracellular Leishmania, this finding demonstrates an important feature of the developed SLN. Collectively, these data indicate the feasibility of developing SLN as potential delivery systems for 17-AAG in leishmaniasis chemotherapy.
  • Autores: Rashidi, S.; Nguewa, Paul (Autor de correspondencia); Mojtahedi, Z.; et al.
    Revista: EASTERN MEDITERRANEAN HEALTH JOURNAL
    ISSN: 1020-3397 Vol.26 N° 12 2020 págs. 1548 - 1555
    Resumen
    Background: In the Mediterranean region, Leishmania infantum is the main cause of visceral leishmaniasis. Dogs with canine visceral leishmaniasis are an important reservoir of visceral leishmaniasis. Control of canine visceral leishmaniasis could disrupt transmission of visceral leishmaniasis to humans. The secreted antigens of Leishmania promastigotes are potential stimuli of the host immune system. Proteomic techniques facilitate the identification of new protein markers. Aims: This study aimed to identify immunoreactive proteins in the secretions of L. infantum promastigotes which could be possible targets for the diagnosis and treatment of canine visceral leishmaniasis and the development of vaccines against the disease. Methods: Secretions of L. infantum promastigotes were obtained from the cultivation of 6 x 10(9) promastigotes in serum-free RPMI-1640 medium during a period of 72 h. After deionization and lyophilization, two-dimensional gel electrophoresis was used for protein separation followed by Western blotting. Thirteen common and repeatable immunoreactive spots were analysed by mass spectrometry. Results: Nine proteins were identified by spectrometry. Most of these proteins were involved in metabolism pathways, survival and pathogenicity of Leishmania parasites. Phospholipase C, immune inhibitor A, chitin-binding protein and a single peptide match to chain A crystal structure of selenomethionine were observed in the secretions of L. infantum promastigotes. Conclusions: The proteins identified in metabolism pathways, survival and pathogenicity of Leishmania parasites are possible targets that could be used for the diagnosis and treatment of canine visceral leishmaniasis and the development of vaccines against the disease in the future.
  • Autores: Rashidi, S.; Kalantar, K.; Nguewa, Paul (Autor de correspondencia); et al.
    Revista: TRANSACTIONS OF THE ROYAL SOCIETY OF TROPICAL MEDICINE AND HYGIENE
    ISSN: 0035-9203 Vol.114 N° 7 2020 págs. 541 - 544
    Resumen
    Optimum Levels of selenoproteins are essential for starting and managing the host immune responses against pathogens. According to the expression of selenoproteins in Leishmania parasites, and since high Levels of selenoproteins Lead to adverse effects on immune cells and their functions, Leishmania parasites might then express selenoproteins such as selenomethionine in their structure and/or secretions able to challenge the host immune system. Finally, this adaptation may Lead to evasion of the parasite from the host immune system. The expression of selenoproteins in Leishmania parasites might then induce the development of infection. We therefore suggest these molecules as new therapeutic candidates for the treatment of leishmaniasis.
  • Autores: Ojo, O. E.; Adebayo, A. S.; Awobode, H. O.; et al.
    Revista: SCIENTIFIC AFRICAN
    ISSN: 2468-2276 Vol.6 2019 págs. e00186
    Resumen
    Malaria and schistosomiasis continue to contribute a big burden to infectious disease prevalence in the tropical areas, mainly in sub Saharan African countries. We previously reported high levels of schistosome specific antibody IgG3 in children coinfected with malaria and schistosomiasis. The aim of the current study was to examine the current co-infection rates of these diseases in Nigeria. Published and unpublished studies on co-infection of human urogenital schistosomiasis and malaria carried out in Nigeria between 2001 and August 2018 were retrieved through literature searches in PubMed, Google Scholar, AJOL, and university theses repositories. The filtered and relevant articles were reviewed and combined in a meta-analysis. Studies involving children reported higher rates of coinfection. The fourteen research articles involving 6,559 individuals were combined in a meta-analysis. Our analyses revealed an estimated 15% co-infection for the country, though with wide variability depending on location. In addition, there are few and well-designed research publications in Nigeria on prevalence and mechanism of malaria and schistosomiasis coinfection.
  • Autores: Fernandez-Rubio, C.; Larrea Leoz, María Esther; Peña Guerrero, J.; et al.
    Revista: ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
    ISSN: 0066-4804 Vol.63 N° 2 2019 págs. e00904-18
    Resumen
    Conventional chemotherapy against leishmaniasis includes agents exhibiting considerable toxicity. In addition, reports of drug resistance are not uncommon. Thus, safe and effective therapies are urgently needed. Isoselenocyanate compounds have recently been identified with potential antitumor activity. It is well known that some antitumor agents demonstrate effects against Leishmania In this study, the in vitro leishmanicidal activities of several organo-selenium and organo-sulfur compounds were tested against Leishmania major and Leishmania amazonensis parasites, using promastigotes and intracellular amastigote forms. The cytotoxicity of these agents was measured in murine peritoneal macrophages and their selectivity indexes were calculated. One of the tested compounds, the isoselenocyanate derivative NISC-6, showed selectivity indexes 2- and 10-fold higher than those of the reference drug amphotericin B when evaluated in L. amazonensis and L. major, respectively. The American strain (L. amazonensis) was less sensitive to NISC-6 than L. major, showing a trend similar to that observed previously for amphotericin B. In addition, we also observed that NISC-6 significantly reduced the number of amastigotes per infected macrophage. On the other hand, we showed that NISC-6 decreases expression levels of Leishmania genes involved in the cell cycle, such as topoisomerase-2 (TOP-2), PCNA, and MCM4, therefore contributing to its leishmanicidal activity. The effect of this compound on cell cycle progression was confirmed by flow cytometry. We observed a significant increase of cells in the G1 phase and a dramatic reduction of cells in the S phase compared to untreated cells. Altogether, our data suggest that the isoselenocyanate NISC-6 may be a promising candidate for new drug development against leishmaniasis.
  • Autores: Vacas Oleas, Andrés Fernando; Fernández Rubio, Celia; Algarabel Olona, Miriam; et al.
    Revista: BIOMOLECULES
    ISSN: 2218-273X Vol.9 N° 11 2019
    Resumen
    The identification and clarification of the mechanisms of action of drugs used against leishmaniasis may improve their administration regimens and prevent the development of resistant strains. Herein, for the first time, we describe the structure of the putatively essential Ser/Thr kinase LmjF.22.0810 from Leishmania major. Molecular dynamics simulations were performed to assess the stability of the kinase model. The analysis of its sequence and structure revealed two druggable sites on the protein. Furthermore, in silico docking of small molecules showed that aminoglycosides preferentially bind to the phosphorylation site of the protein. Given that transgenic LmjF.22.0810-overexpressing parasites displayed less sensitivity to aminoglycosides such as paromomycin, our predicted models support the idea that the mechanism of drug resistance observed in those transgenic parasites is the tight binding of such compounds to LmjF.22.0810 associated with its overexpression. These results may be helpful to understand the complex machinery of drug response in Leishmania.
  • Autores: Vacas Oleas, Andrés Fernando; Fernández Rubio, Celia; Larrea Leoz, María Esther; et al.
    Revista: AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE
    ISSN: 0002-9637 Vol.101 N° Supl. 5 2019 págs. 171
  • Autores: Canovas-García, N.; Díaz Díaz, Roque; López Fidalgo, Jesús Fernando; et al.
    Revista: AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE
    ISSN: 0002-9637 Vol.101 N° Supl. 5 2019 págs. 361 - 361
  • Autores: Leiva León, José; Fernández Alonso, Mirian; Rubio Vallejo, Manuel Francisco; et al.
    Revista: MEDICINE (ELSEVIER)
    ISSN: 0304-5412 Vol.12 N° 50 2018 págs. 2941 - 2951
  • Autores: Fernández Alonso, Mirian; Reina González, Gabriel; Rubio Vallejo, Manuel Francisco; et al.
    Revista: MEDICINE (ELSEVIER)
    ISSN: 0304-5412 Vol.12 N° 49 2018 págs. 2901 - 2909
    Resumen
    Corynebacterium, Listeria and Bacillus are ubiquitous gram-positive aerobic bacilli, which colonize environment, animals and human body. Corynebacterium diphtheriae produces diphtheria either cutaneous or respiratory forms, a disease controlled by vaccination only in some countries and with increased incidence due to immigration and vaccine rejection. Listeria monocytgenes is associated with bacteremia and meningitis in pregnant women, neonates, elderly and immunosuppressed patients, although its incidence at a global level remains unknown. Bacillus anthracis is considered a potential bioterrorism agent because of its lethality. Considering that all cutaneous, gastrointestinal and respiratory symptoms of anthrax could be unspecific, diagnostic procedures that allow their detection with greater safety in case of suspicion have been developed. Toxin production, invasiveness and intracellular survival, resistance in the environment and lytic enzymes production are mechanisms of virulence of these species. Other Corynebacterium or Bacillus species are considered opportunistic pathogens and their clinical relevance has increased along the last decades, with the increase of immunosuppressed patients and the use of medical devices and prostheses susceptible of colonization.
  • Autores: Reina González, Gabriel; Leiva León, José; Rubio Vallejo, Manuel Francisco; et al.
    Revista: MEDICINE (ELSEVIER)
    ISSN: 0304-5412 Vol.12 N° 51 2018 págs. 2991 - 2999
    Resumen
    Introduction.: Tetanus and botulism constitute two life-threating infections caused by spore-forming bacteria, Clostridium tetani and Clostridium botulinum, respectively. Tetanus.: Tetanus is caused by tetanospasmine toxin, which inhibits GABA and glicine neurotransmitters release, causing spastic paralysis followed by respiratory failure and severe impairment of autonomic nervous system. Mortality rate is approximately 8-60%, therefore a prompt clinical diagnosis is essential to transfer the patient to intensive care unit and establish supportive care. In addition, the toxin must be neutralized with specific immunoglobulin, spasms must be controlled with benzodiazepines and cardiovascular instability must be managed with labetalol and magnesium sulphate. Botulism.: Botulism is caused by a thermolabile neurotoxin resulting in flaccid paralysis and respiratory failure, with an observed mortality of 5-10%. There are eight different toxins (A-H) produced by several Clostridium specie, responsible for botulism. Infection can occur following food poisoning with preformed toxin (food-borne botulism), by ingestion of food contaminated with Clostridium botulinum spores (infant botulism), by wound contamination or bacteria inhalation if used as a bioweapon. Early respiratory and airway support must be established together with additional measures (antitoxin and antibiotic).
  • Autores: Peña Guerrero, José; Algarabel Olona, Miriam; Fernández Rubio, Celia; et al.
    Revista: AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE
    ISSN: 0002-9637 Vol.99 N° 4 2018 págs. 176 - 177
  • Autores: Peña Guerrero, José; Puig Rigall, Joan; González Gaitano, Gustavo; et al.
    Libro: Applications of Nanobiotechnology for Neglected Tropical Diseases
    ISSN: 978-0-12-821100-7 2021 págs. 253 - 290
    Resumen
    Leishmaniasis is a neglected tropical disease caused by intracellular parasites from the genus Leishmania. This neglected tropical disease is responsible for significant morbidity and mortality worldwide. One of the most important challenges for the treatment of leishmaniasis is the oral administration of drugs. Such need combined to the predicted high bioavailability of miltefosine prompted its validation. Its antileishmanial activity was better than that of the intravenous sodium stibogluconate. In 2002, it was approved as the first oral drug against leishmaniasis. Miltefosine is being an attractive drug for leishmaniasis due to its cost-effective and oral administration. On the other hand, nanocarriers drug delivery systems used to enhance activity and reduce side effects of antileishmanial drugs. This chapter reviews the most common nanocarriers used for miltefosine, focusing on polymeric micelles, characterized for their core-shell structure. Micelles formed with PEO-PPO block copolymers (Pluronics and Tetronics) and TPGS (PEGylated derivative of vitamin E) are described, as well as their composition, biological activity, and interaction with miltefosine. Promising systems, based on this drug, are reported by the combination of several copolymers, forming polymeric mixed micelles, and the further organization of some PEO-PPO block copolymers micelles into hydrogels.

Proyectos desde 2018

  • Título: Incorporación de fármacos en fibras poliméricas para liberación dérmica: aplicación al tratamiento de leishmaniasis cutánea e infecciones de la piel
    Código de expediente: PID2020-112713RB-C21
    Investigador principal: GUSTAVO GONZALEZ GAITANO, PAUL ALAIN NGUEWA KAMSU.
    Financiador: AGENCIA ESTATAL DE INVESTIGACION
    Convocatoria: 2020 AEI PROYECTOS I+D+i (incluye Generación del conocimiento y Retos investigación)
    Fecha de inicio: 01-01-2021
    Fecha fin: 31-12-2023
    Importe concedido: 84.700,00€
    Otros fondos: -
  • Título: Ayuda para contratar técnologos.
    Código de expediente: 0011-1307-2018-000048
    Investigador principal: PAUL ALAIN NGUEWA KAMSU.
    Financiador: GOBIERNO DE NAVARRA
    Convocatoria: 2018 GN TECNÓLOGOS
    Fecha de inicio: 27-09-2018
    Fecha fin: 26-09-2019
    Importe concedido: 17.615,17€
    Otros fondos: -
  • Título: BÚSQUEDA DE NUEVAS DIANAS Y ESTRATEGIAS TERAPÉUTICAS EN UN MODELO DE LEISHMANIASIS
    Código de expediente: 12/2017
    Investigador principal: PAUL ALAIN NGUEWA KAMSU.
    Financiador: GOBIERNO DE NAVARRA
    Convocatoria: 2017 GN SALUD
    Fecha de inicio: 01-05-2018
    Fecha fin: 30-04-2020
    Importe concedido: 40.020,00€
    Otros fondos: Fondos FEDER
  • Título: Glicoconjugación "click" de nanopartículas inmunoestimulantes y evaluación in vivo del "targeting" selectivo y re-polarización de macrófagos infectados por Leishmania spp.
    Código de expediente: 0011-1383-2018-000005 PI045
    Investigador principal: MARIA SOCORRO ESPUELAS MILLAN.
    Financiador: GOBIERNO DE NAVARRA
    Convocatoria: 2018 GN Centros
    Fecha de inicio: 01-02-2018
    Fecha fin: 30-11-2018
    Importe concedido: 90.422,60€
    Otros fondos: -
  • Título: Validación de nuevas terapias frente a la Leishmaniasis, una patología emergente en Navarra.
    Código de expediente: 0011-1383-2018-000005 PI042
    Investigador principal: PAUL ALAIN NGUEWA KAMSU.
    Financiador: GOBIERNO DE NAVARRA
    Convocatoria: 2018 GN Centros
    Fecha de inicio: 01-02-2018
    Fecha fin: 30-11-2018
    Importe concedido: 96.542,50€
    Otros fondos: -
  • Título: Unravelling Data for Rapid Evidence-Based Response to COVID-19
    Código de expediente:
    Investigador principal: PAUL ALAIN NGUEWA KAMSU
    Financiador: COMISIÓN EUROPEA
    Convocatoria: SC1-PHE-CORONAVIRUS-2020-2E: Networking of existing EU and international cohorts of relevance to COVID-19
    Fecha de inicio: 15-11-2020
    Fecha fin: 14-11-2022
    Importe concedido: 161.750,00€
    Otros fondos: -