id_860. CAMK2B AND SMAP1 AS NOVEL BRAIN PROTEIN BIOMARKERS LINKED TO LONG-TERM ABSTINENCE FOLLOWING EXTENDED COCAINE SELF-ADMINISTRATION
Joanna Jastrzębska1, Saiyara Aniqa1, Kinga Szafran-Pilch2, Anna Drabik3, Renata Pieniążek1, Joanna Wierońska4, Piotr Brański4, Grzegorz Burnat4, Małgorzata Frankowska1, Przemek Mielczarek2,3, Małgorzata Filip1
1 Maj Institute of Pharmacology, Polish Academy of Sciences, Department of Drug Addiction Pharmacology, Kraków, Poland
2 Maj Institute of Pharmacology, Polish Academy of Sciences, Laboratory of Proteomics and Mass Spectrometry, Kraków, Poland
3 AGH University of Krakow, Faculty of Materials Science and Ceramics, Krakow, Poland
4 Maj Institute of Pharmacology, Polish Academy of Sciences, Department of Neurobiology,, Kraków, Poland
INTRODUCTION: Cocaine addiction is a chronic brain disorder and remains a major public health problem, with craving and relapse as core clinical features.
AIM(S): This study aimed to identify brain proteins regulated after prolonged abstinence following extended cocaine self-administration in rats using advanced proteomic approaches.
METHOD(S): Male and female Wistar Han rats were surgically implanted with jugular vein catheters and trained to self-administer cocaine under fixed-ratio schedules (FR1–FR5) with light and sound cues. Animals underwent short (1 h) followed by extended (6 h) access sessions. Yoked cocaine and saline controls were used to dissociate motivational from pharmacological effects. After 30 days of withdrawal in the home cage, the dorsal striatum and prefrontal cortex were collected and analyzed using SP3-based sample preparation and DIA mass spectrometry. Differentially regulated proteins were used to design structure-specific aptamers. A separate group of rats was injected intravenously with Cy5-labeled aptamers, and whole-body fluorescence was quantified by in vivo imaging using the IVIS Lumina III system (Revvity) at 1, 2, and 24 h to assess biodistribution and clearance
RESULTS: Proteomic profiling revealed robust, region-specific protein regulation in relapse-related circuits after prolonged abstinence in both sexes. Sex-dependent targets were identified, with CAMK2B predominant in males and SMAP1 in females. In vivo imaging showed rapid systemic distribution of both aptamers within 1 h, with distinct regional accumulation patterns (CAMK2B, enhanced cranial signal; SMAP1, stronger abdominal signal), followed by partial tissue retention at 2 h and marked signal reduction by 24 h, consistent with progressive clearance
CONCLUSIONS: The study identifies novel, partially sex-specific protein biomarkers associated with prolonged abstinence after cocaine self-administration and demonstrates the feasibility of aptamer-based in vivo target engagement and biodistribution profiling.
FINANCIAL SUPPORT: This research was funded by the Medical Research Agency (ABM) grant number 2024/ABM/03/KPO/KPOD.07.07-IW.07-0104/24.