RNA interference (RNAi) for the silencing of extracellular serine proteases genes in Acanthamoeba: Molecular analysis and effect on pathogenecity

Lorenzo‐Morales, Jacob; Ortega-Rivas, Antonio; Foronda, Pilar; Abreu-Acosta, Néstor; Ballart, David; Martı́nez, Enrique; Valladares, Basilio

doi:10.1016/j.molbiopara.2005.07.001

Cited by 41 publications

(40 citation statements)

References 24 publications

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“…Recently, the application of siRNA in Acanthamoeba species (Lorenzo-Morales et al, 2005;2010) has opened a novel approach for the progress of future therapies based on siRNAs alone or in combination with chemical compounds. Also the use of RNAi moleculaes could be very powerful for the identification of novel drug targets and metabolic pathways in these pathogens that could be exploited for the development of new therapeutic agents.…”

Section: Novel Therapeutic Approachesmentioning

confidence: 99%

Encephalitis Due to Free Living Amoebae: An Emerging Issue in Human Health

Lorenzo‐Morales¹,

Martín-Navarro²,

Martínez-Carretero³

et al. 2011

Non-Flavivirus Encephalitis

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Section: Novel Therapeutic Approachesmentioning

confidence: 99%

Encephalitis Due to Free Living Amoebae: An Emerging Issue in Human Health

Lorenzo‐Morales¹,

Martín-Navarro²,

Martínez-Carretero³

et al. 2011

Non-Flavivirus Encephalitis

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“…RNAi appears promising for silencing gene expression in parasitic pathogens, such as protozoans and helminths, as well as disease vectors, by specific target mRNA interference (9,10,18,20). Analysis of gene functions in pathogens of infectious diseases and their vectors is important for research in drug development, and the silencing effects may be directly employed to control parasite transmission and development.…”

mentioning

confidence: 99%

“…Since it was first applied in the genus Acanthamoeba (9), a few genes involved in Acanthamoeba pathogenesis and the formation of mature cysts have been silenced using siRNA in order to identify them as possible targets for chemotherapy and gene therapy or as key items in cellular functions (4,9,10,13,14). In previous studies, extracellular serine proteases were silenced using siRNAs, resulting in the Acanthamoeba organisms not being able to degrade human corneal cells (9).…”

mentioning

confidence: 99%

“…In previous studies, extracellular serine proteases were silenced using siRNAs, resulting in the Acanthamoeba organisms not being able to degrade human corneal cells (9). More recently, it was established by using siRNAs that serine proteases also play an important role in trophozoite differentiation and reemergence during excystment (4).…”

mentioning

confidence: 99%

“…The soaking method, previously and successfully applied to Acanthamoeba gene silencing (9,10), was used in the study. Regarding the siRNAs, two different molecules that were designed and validated in previous studies (9,10) were used in this work. These molecules target the catalytic domains of extracellular serine proteases (SPsiRNA) and the glycogen phosphorylase gene in Acanthamoeba (GP-siRNA).…”

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confidence: 99%

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Therapeutic Potential of a Combination of Two Gene-Specific Small Interfering RNAs against Clinical Strains of Acanthamoeba

Lorenzo‐Morales

Martín-Navarro

López-Arencibia

et al. 2010

Antimicrob Agents Chemother

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Pathogenic strains of the genus Acanthamoeba are causative agents of severe infections, such as fatal encephalitis and a sight-threatening amoebic keratitis. Antimicrobial therapy for these infections is generally empirical, and patient recovery is often problematic, due to the existence of a highly resistant cyst stage in these amoebae. In previous studies, small interfering RNAs (siRNAs) against the catalytic domains of extracellular serine proteases and glycogen phosphorylase from Acanthamoeba were designed and evaluated for future therapeutic use. The silencing of proteases resulted in Acanthamoeba failing to degrade human corneal cells, and silencing of glycogen phosphorylase caused amoebae to be unable to form mature cysts. After the siRNA design and concentration were optimized in order to avoid toxicity problems, cultures of Acanthamoeba were treated with a combination of both siRNAs, and cells were evaluated under an inverted microscope. This siRNA-based treatment dramatically affected the growth rate and cellular survival of the amoebae. These results were observed less than 48 h after the initiation of the treatment. In order to check possible toxic effects of the siRNA combination, three eukaryotic cell lines (HeLa, murine macrophages, and osteosarcoma cells) were treated with the same molecules, and cytotoxicity was examined by measuring lactate dehydrogenase release. The future use of the combination of these siRNAs is proposed as a potential therapeutic approach against pathogenic strains of Acanthamoeba.

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Guidelines for the purification and characterization of extracellular vesicles of parasites

Fernandez‐Becerra,

Xander,

Alfandari

et al. 2023

J of Extracellular Bio

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Parasites are responsible for the most neglected tropical diseases, affecting over a billion people worldwide (WHO, 2015) and accounting for billions of cases a year and responsible for several millions of deaths. Research on extracellular vesicles (EVs) has increased in recent years and demonstrated that EVs shed by pathogenic parasites interact with host cells playing an important role in the parasite's survival, such as facilitation of infection, immunomodulation, parasite adaptation to the host environment and the transfer of drug resistance factors. Thus, EVs released by parasites mediate parasite‐parasite and parasite‐host intercellular communication. In addition, they are being explored as biomarkers of asymptomatic infections and disease prognosis after drug treatment. However, most current protocols used for the isolation, size determination, quantification and characterization of molecular cargo of EVs lack greater rigor, standardization, and adequate quality controls to certify the enrichment or purity of the ensuing bioproducts. We are now initiating major guidelines based on the evolution of collective knowledge in recent years. The main points covered in this position paper are methods for the isolation and molecular characterization of EVs obtained from parasite‐infected cell cultures, experimental animals, and patients. The guideline also includes a discussion of suggested protocols and functional assays in host cells

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RNA interference (RNAi) for the silencing of extracellular serine proteases genes in Acanthamoeba: Molecular analysis and effect on pathogenecity

Cited by 41 publications

References 24 publications

Encephalitis Due to Free Living Amoebae: An Emerging Issue in Human Health

Encephalitis Due to Free Living Amoebae: An Emerging Issue in Human Health

Therapeutic Potential of a Combination of Two Gene-Specific Small Interfering RNAs against Clinical Strains of Acanthamoeba

Guidelines for the purification and characterization of extracellular vesicles of parasites

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