Preliminary results of the research team. The PL–CZ–CA–US consortium has already generated extensive experimental evidence supporting NUMIEP’s feasibility and conceptual foundation (see references). These studies collectively reconstruct key components of the mechanistic pathway—biofortified food → microbiome → microbial metabolites → host epigenome—providing a strong empirical basis for the integrative research programme outlined in WPs 1–7. The research team has long-standing scientific experience with grants and extensive hands-on expertise with the key instrumentation used in this project, including next-generation sequencing platforms (Illumina/NGS, PL), targeted methylation systems (PyroMark Q48, PL), chromatographic and mass-spectrometric systems (LC–MS/MS, GC–FID, CZ), and advanced cell-culture and organoid 2D/3D models (PL, CZ). This cumulative experience has enabled optimisation of protocols, calibration procedures and analytical workflows, providing a fully validated methodological foundation for implementing all experimental stages of NUMIEP and preliminary results.

A. Plant biofortification and functional-food mechanisms (PL). Koronowicz Aneta, Smoleń Sylweste, Kowalska Iwona. The Polish team has established a complete, validated platform for iodine/selenium biofortification and mechanistic testing of functional lettuce: a) Developed and optimised hydroponic iodine biofortification methods, including uptake kinetics, speciation and biochemical stability [11,26,27]. b) Demonstrated reproducible interactions between iodine, selenium and salicylic acid, confirming predictable metabolic responses essential for WP1 optimisation [28,29,30]. c) Showed that biofortified lettuce extracts modulate inflammatory, metabolic and thyroid-axis markers in epithelial cells [31]. d) Generated reproducible protein/gene-expression datasets (Bio-Plex, qPCR, PyroMark), providing validated pipelines for WP4–WP6. e) Performed dietary intervention studies confirming safety, bioavailability and endocrine relevance of biofortified vegetables [32]. f) Produced baseline microbiome (MiSeq) and promoter-methylation datasets necessary for WP3 and WP5. These results confirm full readiness for WP1–WP2 and provide mechanistic baselines for WP4–WP6 (see references).

B. Microbiome–nutrient–epithelium mechanisms (CZ). Doskočil Ivo., Šmíd Peter. The Czech team contributes experimentally validated intestinal and microbial models essential for WP3-4: Key validated results: a) Validated epithelial systems (Caco-2, HT29-MTX, 3D spheroids) and fermentation platforms capturing mucin secretion, SCFA profiles, bacterial adhesion and metabolite conversions [33,34]. b) Demonstrated diet-dependent modulation of microbial adherence and microbial ecology, including HMOs, prebiotic saccharides and pathogen competition [35,36]. c) Showed that polyphenols, n-3 PUFAs and selenium-enriched probiotics influence immune signalling, barrier function and antioxidant pathways [13,37,38,39]. d) Optimised in-vitro digestion and epithelial integrity workflows [40,41,42]. Together, these results validate feasibility of WP3–WP4 and provide mechanistic anchors for the next step (references). 

C. Epigenomic regulation by dietary molecules (CA). Stefańska Babara (UBC). The Canadian partner has produced high-impact evidence that dietary metabolites reshape the epigenome. Key validated results: a) Demonstrated locus-specific methylation changes, DNMT-dependent silencing, enhancer remodelling and chromatin accessibility shifts induced by dietary stilbenoids and phytoestrogens [43,44,45]. b) Co-authored mechanistic studies showing mitochondrial and epigenetic responses to iodine-biofortified lettuce in epithelial cells [46,47]. c) Provides validated genome-wide methylome and chromatin-analysis pipelines. These findings support WP4–6.

D. Thyroid-axis signalling, RNA biology and multi-omics integration (PL/US). Master Adam. Dr Master contributes an extensive research portfolio spanning the thyroid axis, RNA-mediated regulation and multi-omics integration. He was invited as a worldwide expert to prepare the THRB gene entry for the Atlas of Genetics and Cytogenetics in Oncology and Haematology (2014), and his mechanistic publications are cited in the NCBI THRB gene description. His scientific awards and recognised expertise provide a strong mechanistic foundation for NUMIEP’s thyroid-axis analyses in WP2 and WP5–WP7 (see his refenreces). Key validated results: a) Validated mechanistic baseline of transcriptional and translational control of TH-THRB/THRA. b) Identified and experimentally characterised novel THRB 5′/3′ UTR isoforms [2]. c) Demonstrated specific regulation of THRB that is controled not only by promoter CpG hypermethylation, but also microRNA-mediated post-transcriptional silencing [3]. d) Developed a novel method for gene specific enhancement of protein translation by targeting 5’UTRs of selected tumor suppressors including THRB that will be useful in WP6 [48]. e) Provided validated multi-omics pipelines and protocols for genome, transcriptome and proteome analyses [49,50]. f) Generated preliminary murine datasets demonstrating diet-responsive modulation of THRB/THRA, DNMT/TET networks and histone modifiers, central to WP5-7.

E.Integrative preliminary evidence from joint publications (an example of PL–CA collaboration). The joint PL–CA study showed that iodine-biofortified lettuce modulates mitochondrial function, oxidative-stress responses, cell-cycle progression and epigenetic regulation in human epithelial cancer models. Organic iodine–enriched extracts reduced HT-29 cell viability and proliferation, induced S-phase arrest, mitochondrial depolarization and cytochrome-c–dependent apoptosis. Lettuce fortified with 5-ISA induced promoter hypomethylation and ~2-fold upregulation of the tumor suppressor SEMA3A, consistent with DNMT3A-linked methylation control and RB/MDM2 interactions. While THRB regulates DNMT3A within a thyroid-related epigenetic loop, SEMA3A represents a distinct class of microbiota- and SCFA-responsive genes governed by promoter methylation and HDAC-dependent chromatin remodeling. Control lettuce also induced SEMA3A hypomethylation, with effects amplified by 5-ISA biofortification, reinforcing the NUMIEP framework. These findings provide mechanistic evidence that iodine-biofortified functional foods elicit epigenetically mediated metabolic and transcriptional responses, supporting the feasibility of WP4–WP6.