P1.24. SPNAR9810Q MICE:
DEEP BEHAVIORAL AND MOLECULAR PHENOTYPING
OF SPONTANEOUS SPECTRINOPATHY MODEL
Patrycja Ziuzia1,2, Dzesika Hoinkis3, Monika Opałek4, Michał Ślęzak1, Arkadiusz Miążek2
1 Łukasiewicz Research Network - PORT Polish Center for Technology Development, Life Sciences and Biotechnology Center, Stabłowicka 147, Wrocław, Poland
2 Wroclaw University of Environmental and Life Sciences, Department of Biochemistry and Molecular Biology, Norwida 25, Wrocław, Poland
3 Maj Institute of Pharmacology Polish Academy of Sciences, Department of Molecular Neuropharmacology, Smętna 12, Kraków, Poland
4 Intelliseq Inc., Wilczyńskiego 19, Kraków, Poland
INTRODUCTION: Progressive neuronal loss or proteostasis disruption underlying eurodegenerative disorders are often linked to dysregulated calpain activity. A subtype of these conditions are spectrinopathies – disorders associated with hereditary cerebellar ataxias – caused by mutations in SPTAN1 (non-erythrocytic α-II-spectrin). In previous studies, we discovered a spontaneous missense mutation in SPTAN1 gene (Sptan1 c.3293G > A:p.R1098Q) causing premature neurodegeneration in affected individuals.
AIM(S): We hypothesized that expressing human calpastatin, an endogenous calpain inhibitor, can mitigate molecular and behavioral alterations associated with the mutation.
METHOD(S): To test the hypothesis, we generated a transgenic line carrying both, the SPTAN1 mutation and human calpastatin gene (Spna2R1098Q x hCAST line). To profile gene expression in affected cerebellar tissue, total RNA was isolated and subjected to Next-Generation Sequencing, followed by differential expression profiling. Functional enrichment (e.g. GO, KEGG) and protein-protein interactions analysis were performed to identify affected biological pathways and prioritize candidate genes for further investigation. To detect subtle, mutation-driven phenotypic differences throughout developmental stages, we apply automated mice tracking
in semi-naturalistic conditions using open-source tools for animal tracking (SLEAP.ai) and behavior interpretation (DeepOF). Cognitive performance are evaluated via Novel Object Recognition Test (EthoVision XT15, Noldus).
RESULTS: Our findings provide pioneer transcriptomic profile of the Spna2R9810Q mouse model. Although, we did not detect statistically significant impact of co-expressed hCAST on transcriptomic profile, 8 gene candidates were targeted for further investigation of their potential involvement in pathogenesis of SPTAN1 ataxia.
CONCLUSIONS: Ongoing behavioral studies aim to explore whether SPTAN1 mutation affects not only motor abilities, but also social behavior – an aspect not yet explored in this model.
FINANCIAL SUPPORT: This project is a part of Industrial PhD Programme: VI Edition, Ministry of Science and Higher Education, Poland (grant nb: DWD/6/00191/2022) and a part of National Science Centre grant, based on decision no. Dec-2021/41/B/NZ3/04099, entitled “Do astrocytes control synaptic connections in neural networks relevant to psychiatric diseases?”.