Integrated Transcriptomic Profiling and In Vitro Evaluation of Garcinia Mangostana Phytochemicals Against...

Cryptosporidium parvum is a major cause of severe diarrheal disease, yet therapeutic options remain limited. This study integrated host-parasite transcriptomic profiling, GC-MS phytochemical profiling, computational screening, and in vitro experimental validation to evaluate anti-Cryptosporidium phytochemicals from Garcinia mangostana extract (GME). Host epithelial transcriptional responses (GSE2077) during C. parvum infection were analyzed to map infection-associated host reprogramming. A curated library of 30 identified phytochemicals was computationally docked against prioritized parasite-relevant targets: calcium-dependent protein kinase 1 (CDPK1), C. parvum 1-phosphatidylinositol 4-kinase [PI(4)K; cgd8_4500/UniProt Q5CVD3, AlphaFold model AF-Q5CVD3-F1], and cleavage and polyadenylation specificity factor 3 (CPSF3). The anti-Cryptosporidium efficacy of GME was subsequently evaluated in vitro by assessing C. parvum oocyst count and viability over 48 h. Transcriptomic analysis suggested a biphasic pattern of host reprogramming involving inflammatory signaling, sterol metabolism, Wnt signaling, and cell-cycle regulation. GC-MS analysis identified sterols and fatty acid derivatives, including stigmasterol, campesterol, and ethyl iso-allocholate. Molecular docking against the C. parvum PI(4)K model predicted campesterol (NP028) and ethyl iso-allocholate (NP029) as leading PI(4)K-oriented candidates, with mean affinities of - 7.97 ± 0.59 and - 7.72 ± 0.61 kcal/mol, respectively, while NP008 showed a predicted multi-target interaction profile, particularly against CPSF3. In vitro assays showed a significant, concentration- and time-dependent reduction in C. parvum oocyst viability, with an LC50 of 830 µg/mL after 48 h. Overall, the parasite-specific docking and oocysticidal data support GME phytochemicals as promising anti-Cryptosporidium candidates and generate testable hypotheses for CDPK1, PI(4)K, or CPSF3 involvement, while direct intracellular infection assays and recombinant enzyme inhibition studies are required to confirm target engagement.