Abstract
Availability of living organisms to mimic key step of amyloidogenesis of human protein has become an indispensable tool for our translation approach aiming at filling the deep gap existing between the biophysical and biochemical data obtained in vitro and the pathological features observed in patients. Human β2-microglobulin (β2-m) causes systemic amyloidosis in haemodialysed patients. The structure, misfolding propensity, kinetics of fibrillogenesis and cytotoxicity of this protein, in vitro, have been studied more extensively than for any other globular protein. However, no suitable animal model for β2-m amyloidosis has been so far reported. We have now established and characterized three new transgenic C. elegans strains expressing wild type human β2-m and two highly amyloidogenic isoforms: P32G variant and the truncated form ΔN6 lacking of the 6 N-terminal residues. The expression of human β2-m affects the larval growth of C. elegans and the severity of the damage correlates with the intrinsic propensity to self-aggregate that has been reported in previous in vitro studies. We have no evidence of the formation of amyloid deposits in the body-wall muscles of worms. However, we discovered a strict correlation between the pathological phenotype and the presence of oligomeric species recognized by the A11 antibody. The strains expressing human β2-m exhibit a locomotory defect quantified with the body bends assay. Here we show that tetracyclines can correct this abnormality confirming that these compounds are able to protect a living organism from the proteotoxicity of human β2-m.
Highlights
Caenorhabditis elegans is an extremely versatile and appropriate animal model for mimicking and recapitulating in vivo the key molecular mechanisms underlying the gene-and tissue-specific protein misfolding and toxicity related to the human pathogenesis [1]
Quantification of the immunoreactive dots indicated that both wild type b2-m (WT) and P32G transgenic strains expressed comparable b2-m levels (0.3260.04 and 0.3460.06 density/mg protein for WT and P32G, respectively) whereas a lower, but not statistically significant, protein content was detected in DN6 animals (0.1960.05 density/mg protein) (Figure 2B)
We report the first model of transgenic C. elegans expressing and directing human b2-m in the muscular system
Summary
Caenorhabditis elegans is an extremely versatile and appropriate animal model for mimicking and recapitulating in vivo the key molecular mechanisms underlying the gene-and tissue-specific protein misfolding and toxicity related to the human pathogenesis [1]. Despite the evolutionary distance from vertebrates, human proteins substantially maintain their structure and function when they are expressed in C. elegans [1]. Many variant proteins associated to human diseases cause a pathological phenotype in worms and this cross-species translation greatly facilitates the study of human diseases in this simple organism. This is true for ‘‘gain of function diseases’’, including Alzheimer, Parkinson and Huntington diseases, caused by self-aggregation of specific peptides [2,3,4]. Transgenic worms expressing human diseaserelevant proteins and peptides represented a rapid and highly informative system for the screening of putative therapeutic medications at the early stages of drug development with particular regard to aging-related diseases [5]. Alavez et al [6]
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