Emerging biomarkers in amyotrophic lateral sclerosis: from pathogenesis to clinical applications

Abstract

Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative condition marked by the gradual loss of motor neurons in the brain and spinal cord. As the most common adult-onset motor neuron disease, ALS manifests through gradually worsening muscle weakness that ultimately progresses to complete paralysis. The disease presents in both sporadic and familial forms. Diagnosis is often delayed until substantial and irreversible motor neuron damage has already occurred. Clinical outcomes in ALS have only been defined through large-scale clinical trials with lengthy follow-up periods due to the disease's inherent heterogeneity and the absence of disease-specific biomarkers. Current biomarker detection methods, such as invasive cerebrospinal fluid (CSF) analysis or advanced imaging, are impractical for routine use, particularly in late-stage ALS. Several blood-based biomarkers have shown promise, including neurofilament levels, cryptic RNA-derived peptides, and immune-mediated changes, which may enable non-invasive monitoring. Nevertheless, the development of these methods is hindered by technical challenges, such as blood matrix interference and low analyte abundance. Among the emerging biomarkers, neurofilament light chain (NfL) appears to be the most promising, as its concentrations change in line with disease progression and distinguish clinically relevant groups. NfL facilitates patient stratification based on clinical progression rates (e.g., rapid vs slow progressors), while cryptic exon-derived peptides, such as UNC13A-derived peptides, enable genetic stratification by identifying molecular subtypes linked to TDP-43 pathology (e.g., C9orf72 vs sporadic ALS). These biomarkers hold promise to optimize clinical trial design through enriched cohort selection and accelerating therapeutic translation by monitoring target engagement. In this review, we have summarized recent developments in ALS biomarker studies, focusing on neurofilaments in each biofluid, transcriptomic signatures, and neuroinflammatory biomarkers, emphasizing technical challenges surrounding reproducibility in measurement. Finally, we discussed the potential integration of these biomarkers into clinical practice to advance drug development through precision medicine, thereby enabling shorter and more targeted clinical trials.

Keywords: amyotrophic lateral sclerosis; biomarkers; molecular basis of neurodegeneration; neuroinflammation markers; therapeutic targets.

FIGURE 1

Pathogenic factors and their interactions in amyotrophic lateral sclerosis (ALS). The ALS pathogenesis arises from the interplay of genetic, molecular, and systemic mechanisms. Genetic mutations disrupt RNA processing and protein homeostasis, leading to toxic aggregates that impair mitochondrial function, elevate reactive oxygen species (ROS), and compromise autophagy. Concurrently, defects in axonal transport deprive neurons of critical organelles and proteins, accelerating degeneration. Emerging evidence implicates gut dysbiosis via the gut-brain axis in amplifying neuroinflammation and oxidative stress, further disrupting neuronal integrity. These pathways collectively drive motor neuron loss through interconnected cycles of cellular dysfunction. Preclinical and clinical studies empirically support most of the mechanisms shown, though contributions from gut dysbiosis remain under active investigation. This illustration is inspired by Jiang and Xu (2025).

FIGURE 2

Mechanisms of cryptic exon inclusion in ALS and detection of associated biomarkers. TDP-43 mislocalization disrupts RNA splicing, leading to the incorporation of cryptic exons into transcripts of genes such as UNC13A and STMN2. These aberrantly spliced mRNAs produce truncated proteins with cryptic peptides derived from intronic sequences. Detection of these peptides in biofluids (e.g., CSF, serum) via mass spectrometry offers a novel biomarker strategy for ALS diagnosis and monitoring therapeutic interventions targeting RNA processing in ALS. This schematic integrates both experimentally validated pathways and emerging mechanisms currently under validation in ALS cohorts.