mHTT Aggregation/Lowering Assay

Huntington's disease (HD) is a common genetic neurodegenerative disorder with a characterization of uncontrolled chorea hyperkinesia, cognitive impairment, and emotional deficits. Much evidence suggests that the pathogenesis of HD is caused by mutations in the Huntingtin (HTT) gene and abnormal expansion and aggregation of the HTT protein. With the service of mHTT aggregation/lowering assay, Creative Biolabs is your best choice for an in-depth study of the pathogenic mechanism of mHTT and to advance your research further.

mHTT Aggregation

As the most prominent pathological phenomenon of HD, the misfolding and abnormal aggregation of mHTT in neurons may be an important factor leading to the pathogenesis and even driving the disease process. Expansion of CAG repeats in polyglutamine at the N-terminus of HTT produces a longer polyQ derivative that self-assembles with mHTT fragments into highly stable, β-sheet-rich protein aggregates. This aggregate has a fibrillar morphology and is highly prone to intracellular aggregation, resulting in significant damage and cell loss in regions of the brain such as the striatum, cortex, etc.

mHTT Lowering

Modulating mHTT levels in brain regions or reducing intracellular mHTT accumulation levels may be the most promising approaches for HD treatment development. Various mHTT lowering agents have been developed. Antisense oligonucleotides and small interfering RNAs were shown to effectively inhibit mHTT expression, and adeno-associated virus-mediated miRNA delivery also significantly reduced aggregate levels. A relatively new area is the application of zinc-finger arrestin proteins that selectively target mutant HTT, which also reduces mHTT aggregate content and improves behavioral deficits in several mouse models. In addition, some chemical agents and derivatives have also been developed to reduce the total amount of mHTT in cells or the aggregate content in neurons by regulating phosphatidylinositol balance or increasing autophagy levels.

Fig 1. Potential toxic species in HD and mechanisms for HTT lowering. (Sarah, et al., 2019)

mHTT Aggregation/Lowering Assay Services at Creative Biolabs

Several in vitro and in vivo models have been developed for mHTT research, we currently offer animal models including R6/2, zQ175 knock-in and BACHD mouse models, as well as commonly used cell models like fibroblasts, StHdh cells, PC12 cells, etc. In addition to inducing genetic mutations, aggregation of mHTT can also be achieved by introducing an MBP tag at the N-terminus of the HTT construct. The introduction of MBP can keep the protein in a soluble state, and then the thrombin protease cleaves the MBP-HTT junction site in solution to directly cause HTT aggregation. Commonly used methods for the quantitative determination of HTT include enzyme-linked immunosorbent assay platforms or methods developed on the basis of electrochemiluminescence that are dependent/independent of the length of the polyglutamine. In addition to the above methods, we also provide common immunological assays, fluorescent staining analysis, and mass spectrometry testing services.

Fig 2. Fluorescent staining analysis of mHTT aggregation. (Perez-Arancibia, et al., 2021)

The single-gene disease nature of HD makes the study of mHTT aggregation/lowering more attractive. The aggregation of HTT protein can manufact a thorough and realistic model of HD, and operations to alleviate protein misfolding or lowering misfolded proteins often cumber the disease progression in HD models. Creative Biolabs can provide accurate, comprehensive and repeatable mHTT aggregation/lowering assays. If you want to advance your experiments or need HD in vitro models and perform tests in order to solve the mystery of HD, please contact us for our professional and efficient R&D team.

References

1. Sarah, J.T.; et al. Huntingtin Lowering Strategies for Disease Modification in Huntington’s Disease. Neuron. 2019, 101: 801-819.
2. Perez-Arancibia, R.; et al. A phenolic-rich extract from Ugni molinae berries reduces abnormal protein aggregation in a cellular model of Huntington’s disease. PLoS ONE. 2021 16(1): e0254834.