Amyotrophic Lateral Sclerosis Biomarker Assays

Hyperexcitation Assay ALS iPSC Cells

Imaging

We are continuously testing and developing new biomarker assays in the field of amyotrophic lateral sclerosis to support:

our validation of our disease models,
for additional counter-screening assays,
to improve predictivity in screening campaigns,
and to develop single assays to prescreen at lower costs.

We have performed a wide range of assays:

Viability Assays

Viability assays can be used to detect the potential capacity of a compound to prolong the survivability of cells. These assays can be combined with toxic agents or cells, such as pathological astrocytes, to amplify effects.

Toxilight

The Toxilight assay measures the adenylate kinase of damaged cells in supernatants. This is an easy and simple marker that can emphasize your results. We perform this assay in most projects as a control experiment. This assay can easily be combined with the MEA readout.

CellTiterGlo

The Cell Titer Glo assay measures the quantity of ATP in lysates and correlates with the number of living cells. This assay can be performed with cells sourced from a MEA experiment.

Live Dead Assay

With the live-dead assay, the ratio of living cells to dead cells can be precisely determined. This assay requires well plates capable of imaging.

Inflammation Assays

Inflammation is supposed to play a role in amyotrophic lateral sclerosis. In co-culture experiments with astrocytes and/or microglia, inflammatory mechanisms can be modeled.

IL1β

The blockade of IL1β is a potential target to stop ALS. The assay can be combined with MEA experiments.

Neurofilament Light Chain Assay

Neurofilament light chain is a diagnostic biomarker for ALS with blood samples. Therefore, an NF-L assay is an important biomarker that mimics clinical symptoms in a dish. Interestingly, the localization of NF-L at the initial axonal segment is connected to hyperexcitation.

NF-L

The secretion of NF-L can be measured along with MEA experiments from supernatants.

Protein mislocalization or aggregation

Proteinopathies are the most discussed cornerstone of disease mechanisms in ALS.

TDP-43 mislocalization

TDP-43 mislocalization can be detected in disease cell lines of ALS models and quantitively assessed with our microscopic approaches.

Further ALS-relevant biomarkers are aberrant RNA homeostasis-causing stress granules or, in C9orf72 cells, dipeptide repeat proteins (DPR), which can be detected and quantified microscopically.

Target Validation with RNA Sequencing

We have conducted comprehensive transcriptomic analyses on our ALS cell lines. If you have a specific gene of interest, feel free to reach out to us for validation of its expression.

Energy Metabolism

Our research investigates the role of energy metabolism using transcriptomic data from iPSC-derived cell cultures of both ALS patients and healthy individuals. This approach serves as a crucial tool for assessing the effects of various compounds. We have specifically analyzed energy metabolism in C9orf72 and SOD1 cell lines, revealing significant variability that aligns with existing literature.

Given the hyperexcitation observed in our diseased cultures, we conduct thorough evaluations of potential dysfunctions in energy metabolism. This comprehensive analysis provides a consistent understanding of how compounds impact ATP, pyruvate, branched-chain amino acids (BCAAs), fatty acid metabolism, and more. Our studies on energy metabolism offer valuable insights for developing therapeutic dietary strategies in ALS for example.

For examples and additional biomarker assays, please don’t hesitate to contact us.