Fabry Disease Animal Model Service
Fabry disease is an X-linked lysosomal storage disorder caused by deficiency of the enzyme α-galactosidase A (α-Gal A), leading to progressive accumulation of globotriaosylceramide (Gb3 or GL-3) and related glycosphingolipids in cells throughout the body, particularly in the vasculature, kidneys, heart, and nervous system. Protheragen provides customized Fabry disease animal model services tailored to your drug discovery and preclinical development needs, including model generation, characterization, pharmacokinetic and pharmacodynamic evaluations, efficacy studies, and safety assessments across multiple therapeutic modalities.
Overview of Fabry Disease Animal Models
Fabry disease is the second most common lysosomal storage disorder, resulting from loss-of-function mutations in the GLA gene that encodes α-galactosidase A. Deficiency of this lysosomal enzyme causes systemic accumulation of glycosphingolipid substrates, most notably globotriaosylceramide (Gb3) and its deacylated form globotriaosylsphingosine (lyso-Gb3). The progressive accumulation of these substrates occurs in vascular endothelial cells, renal podocytes and tubules, cardiac myocytes, dorsal root ganglia neurons, and other tissues, driving the multisystemic pathology characteristic of the disease.
Fig.1 Experimental design for AAV-mediated therapy in GLA-KO mice. (Hayashi, Y., et al., 2023)
Developing clinically relevant animal models is essential for mimicking this systemic pathology. These preclinical models serve as indispensable tools for understanding downstream pathogenic mechanisms, identifying novel biomarkers, and validating the in vivo efficacy, pharmacokinetics, and safety of emerging therapeutics before they enter human trials. Among the most widely used platforms is the GLA knockout mouse, which recapitulates the core enzyme deficiency and substrate accumulation seen in human individuals.
Our Services
Building on deep expertise in rare disease research and transgenic model technologies, Protheragen's team offers comprehensive, custom-designed Fabry disease animal model services across the entire preclinical continuum, from model selection and characterization to full-service drug efficacy evaluation, pharmacokinetic profiling, and IND-enabling safety assessments.
Animal Models of Fabry Disease
Leveraging our advanced genetic engineering platforms and deep insights into lysosomal storage disorders, we provide bespoke development and characterization services to establish the precise in vivo system required for your therapeutic mechanism of action. Each model is generated on demand, with control over genetic background, mutation type, and transgene expression. All custom models undergo rigorous characterization including enzyme activity assays, substrate quantification (Gb3 and lyso-Gb3), histopathological evaluation of target organs (kidney, heart, and nervous tissue), and behavioral phenotyping where relevant.
Custom development of GLA-deficient models enables precise control over genetic background, mutation type, and transgene expression. These models are generated using established gene-targeting and transgenic strategies, allowing for rigorous characterization of enzyme activity, substrate burden, and downstream pathology. Below are representative model types available for custom development.
- GLA-KO Mouse Model: Complete knockout of the endogenous Gla gene resulting in absent α-Gal A activity.
- G3S/GLA-KO Model: Double modification combining GLA knockout with Gb3 synthase overexpression for aggravated substrate burden and accelerated pathology.
- GLA Transgenic Model: Customized to express humanized GLA transgene variants carrying specific clinical missense mutations.
Mouse Model for Fabry Disease Research
| Model Name | Gla-KO Mouse |
|---|---|
| Model Type | Genetically Engineered Mouse Model (GEMM) |
| Modeling Method | Knockout |
| Targeted Disease | Fabry Disease |
| Sales Status | Repository Live |
| Detailed Description | This model features a targeted deletion of exons 2 and 3 within the endogenous Gla gene, completely disrupting α-galactosidase A transcription and enzymatic activity. |
| Applications & Therapeutic Areas | Evaluation of novel therapies for Fabry disease, including enzyme replacement therapies (ERTs), substrate reduction therapies (SRTs), and gene therapies; investigation of lysosomal storage mechanics, pathological lipid accumulation, and associated peripheral sensory neuropathies. |
Case Study-Gla-KO Mouse Model Development
Model Introduction
The development of a predictive preclinical model is essential for evaluating therapeutic interventions against Fabry disease, an inherited lysosomal storage disorder driven by α-galactosidase A deficiency. To fulfill this need, a targeted α-galactosidase A knockout (Gla-KO) mouse model was engineered to serve as a robust platform for evaluating in vivo drug efficacy, pharmacokinetic profiles, and safety. This model reliably mimics the fundamental molecular deficits and neurological clinical manifestations observed in human individuals, providing a validated system for translational research.
Methodology
- Animal Model: The Gla-KO mouse line was successfully generated by employing targeted genomic editing to delete exons 2 and 3 of the endogenous murine Gla gene, effectively preventing functional protein synthesis. All animals were maintained under specific pathogen-free conditions with standard rodent chow and water provided ad libitum.
- Phenotypic Analysis Methods: To confirm genomic disruption and subsequent loss of transcription, brain tissue samples were harvested from mature mice and subjected to quantitative PCR (qPCR) assays utilizing specific primers targeting murine Gla. Phenotypic characterization was conducted on 14-week-old male homozygous Gla-KO mice to evaluate biochemical biomarkers and sensory nerve function. Systemic biochemical analysis was performed by measuring total α-galactosidase A (GLA) enzymatic activity and quantifying pathogenic globotriaosylsphingosine (lyso-Gb3) levels in collected serum samples. To assess functional peripheral neuropathy and thermal sensory deficits, mice were subjected to hot plate behavioral testing, where thermal response latencies were recorded and analyzed.
Phenotypic Analysis & Results
Quantitative PCR analysis of brain tissue was performed to assess residual Gla transcript levels in the custom-developed knockout model. Following RNA extraction and reverse transcription, qPCR was conducted using primer pairs specific to the murine Gla gene. The results demonstrated that Gla mRNA expression in homozygous Gla-KO mice was reduced to near-background levels compared to wild-type controls, with a statistically significant decrease observed across multiple independent samples. These data confirmed successful disruption of the Gla gene at the transcriptional level, validating the intended loss-of-function genotype in the custom-developed knockout model.
Fig.2 Absence of Gla gene expression in the brain of Gla-KO mice as determined by qPCR. Data are presented as mean ± SEM (n=5; ***p < 0.001).
Biochemical analysis confirmed that the genetic deletion successfully translated into systemic enzymatic deficiency and metabolic accumulation. Serum samples analyzed from 14-week-old male Gla-KO mice revealed a profound reduction in functional GLA enzyme activity compared to wild-type controls. Concurrently, a significant elevation of serum lyso-Gb3 levels was detected, confirming systemic lysosomal storage pathology.
Fig.3 Reduced serum α-Gal A activity and elevated lyso-Gb3 in Gla-KO mice at 14 weeks of age. Data are presented as mean ± SEM (n=5; ***p < 0.001).
Behavioral evaluations confirmed that metabolic storage pathology led to functional somatosensory deficits. During hot plate sensory testing, 14-week-old male Gla-KO mice exhibited significantly prolonged response latencies when exposed to thermal stimuli compared to the control group. This marked thermal hypoalgesia demonstrated the presence of functional small-fiber peripheral neuropathy, which mirrors aspects of the sensory abnormalities reported in Fabry disease individuals (altered thermal perception and pain processing).
Fig.4 Hypoalgesic thermal response in 14-week-old male Gla-KO mice on the hot plate test. Knockout cohorts exhibit significantly increased thermal pain thresholds and delayed reaction times, confirming the development of functional peripheral sensory deficits. Data are presented as mean ± SEM (n=5; *p < 0.05).
Conclusion
The custom-developed Gla-KO mouse model successfully recapitulated the key molecular, biochemical, and behavioral features of Fabry disease. Deletion of exons 2-3 resulted in complete loss of Gla expression, near-undetectable serum α-Gal A activity, substantial accumulation of the disease biomarker lyso-Gb3, and a measurable hypoalgesic phenotype on the hot plate test. This well-characterized model provides a reliable platform for evaluating enzyme replacement, gene therapy, and small-molecule candidates targeting Fabry disease, with validated endpoints for both efficacy and peripheral neuropathy assessment.
Contact Us
Beyond Fabry disease animal model generation and characterization, Protheragen's integrated platform provides full-spectrum preclinical development support: pharmacokinetic assessments; comprehensive drug safety evaluation including general toxicology and IND-enabling studies; efficacy testing across multiple therapeutic modalities (enzyme replacement, gene therapy, pharmacological chaperones, substrate reduction therapy, and mRNA-based approaches); as well as customized biomarker analysis and histopathological evaluation of substrate accumulation in cardiac, renal, neural, and vascular tissues. For detailed information, a customized proposal tailored to your specific program needs, or to discuss ongoing collaborations, please contact our scientific team.
Reference
- Hayashi, Yuka et al. "Therapeutic strategy for Fabry disease by intravenous administration of adeno-associated virus 2 or 9 in α-galactosidase A-deficient mice." The journal of gene medicine 25.12 (2023): e3560.
All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.