Breast cancer (BRCA) malignancy causes major fatalities amongst women worldwide. SCF (Skp1-cullin-F-box proteins) E3 ubiquitin ligases are the most well-known members of the ubiquitination–proteasome system (UPS), which promotes cancer initiation and progression. Recently, we demonstrated that FBXL8, a novel F-box protein (SCF^F-boxes) of SCF E3 ligase, accelerates BRCA advancement and metastasis. Since SCF^F-boxes is a key component of E3 ligases, we hypothesized that other SCF^F-boxes besides FBXL8 probably collaborate in regulating breast carcinogenesis. In this study, we retrospectively profiled the transcriptome of BRCA tissues and found a notable upregulation of four SCF^F-box E3 ligases (FBXL8, FBXO43, FBXO15, and CCNF) in the carcinoma tissues. Similar to FBXL8, the knockdown of FBXO43 reduced cancer cell viability and proliferation, suggesting its pro-tumorigenic role. The overexpression of CCNF inhibited cancer cell progression, indicating its anti-tumorigenic role. Unexpectedly, CCNF protein was markedly downregulated in BRCA tissues, although its mRNA level was high. We showed that both E3 ligases, FBXL8 and FZR1, pulled down CCNF. Double knockdown of FBXL8 and FZR1 caused CCNF accumulation. On the other hand, CCNF itself pulled down a tumorigenic factor, RRM2, and CCNF overexpression reduced RRM2. Altogether, we propose a signature network of E3 ligases that collaboratively modulates CCNF anti-cancer activity. There is potential to target BRCA through modulation of the partnership axes of (i) CCNF-FBXL8, (ii) CCNF-FZR1, and (iii) CCNF-RRM2, particularly, via CCNF overexpression and activation and FBXL8/FZR1 suppression.

NSCLC (non-small cell lung cancer) is a leading cause of cancer-related deaths worldwide. Clinical trials showed that Hiltonol, a stable dsRNA representing an advanced form of polyI:C (polyinosinic-polycytidilic acid), is an adjuvant cancer-immunomodulator. However, its mechanisms of action and effect on lung cancer have not been explored pre-clinically. Here, we examined, for the first time, how a novel Hiltonol cocktail kills NSCLC cells. By retrospective analysis of NSCLC patient tissues obtained from the tumor biobank; pre-clinical studies with Hiltonol alone or Hiltonol⁺⁺⁺ cocktail [Hiltonol+anti-IL6+AG490 (JAK2 inhibitor)+Stattic (STAT3 inhibitor)]; cytokine analysis; gene knockdown and gain/loss-of-function studies, we uncovered the mechanisms of action of Hiltonol⁺⁺⁺. We demonstrated that Hiltonol⁺⁺⁺ kills the cancer cells and suppresses the metastatic potential of NSCLC through: (i) upregulation of pro-apoptotic Caspase-9 and Caspase-3, (ii) induction of cytosolic cytochrome c, (iii) modulation of pro-inflammatory cytokines (GRO, MCP-1, IL-8, and IL-6) and anticancer IL-24 in NSCLC subtypes, and (iv) upregulation of tumor suppressors, PKR (protein kinase R) and OAS (2′5′ oligoadenylate synthetase). In silico analysis showed that Lys296 of PKR and Lys66 of OAS interact with Hiltonol. These Lys residues are purportedly involved in the catalytic/signaling activity of the tumor suppressors. Furthermore, knockdown of PKR/OAS abrogated the anticancer action of Hiltonol, provoking survival of cancer cells. Ex vivo analysis of NSCLC patient tissues corroborated that loss of PKR and OAS is associated with cancer advancement. Altogether, our findings unraveled the significance of studying tumor biobank tissues, which suggests PKR and OAS as precision oncological suppressor candidates to be targeted by this novel Hiltonol⁺⁺⁺ cocktail which represents a prospective drug for development into a potent and tailored therapy for NSCLC subtypes.

Background: Population aging is driving a convergent rise in three disorders historically managed in isolation: cardiovascular disease, neurocognitive decline, and osteoporotic bone loss. Accumulating mechanistic data indicate that these systems are coupled through shared regulators of calcium trafficking, inflammation resolution, vascular integrity, and inflammaging, supporting the concept of an integrated "Heart-Brain-Bone" axis. Three lipid-soluble nutrients-long-chain omega-3 polyunsaturated fatty acids (EPA/DHA), vitamin D3 (cholecalciferol), and vitamin K2 (menaquinone-7 [MK-7])-act on overlapping nodes of this network. Methods: We conducted a structured narrative review of mechanistic, observational, and randomized controlled trial (RCT) evidence, grading the strength of support for each claim. Results: Vitamin K2-dependent gamma-carboxylation of matrix Gla protein (MGP) and osteocalcin is proposed to reciprocally direct calcium away from the arterial wall and toward the bone matrix, offering a candidate mechanistic resolution of the "calcium paradox" associated with isolated vitamin D3 supplementation; EPA/DHA-derived specialized pro-resolving mediators may support resolution of endothelial and neuronal inflammation; and bone-, vascular-, and brain-derived signals (osteocalcin, FGF23, the neurovascular unit) interconnect the three organs. Human clinical evidence, however, is heterogeneous and formulation- and population-dependent: cardiovascular omega-3 RCTs are discordant (REDUCE-IT, which used icosapent ethyl [an EPA ethyl ester], positive; VITAL/STRENGTH/ASCEND null, predominantly in lower-risk or replete cohorts), cognitive trials are largely null or subgroup-dependent (MAPT, DO-HEALTH), and MK-7 improves surrogate bone and calcification biomarkers, with only a single recent hard-endpoint coronary calcification RCT (VitaK-CAC). Conclusions: The combined use of long-chain omega-3, vitamin D3, and MK-7 is mechanistically rational and hypothesis-generating rather than clinically established; benefit appears most plausible in individuals with elevated risk or demonstrable nutritional insufficiency, and least in replete, low-risk populations. Findings should be interpreted within a broader healthy-aging context that includes lifestyle and psychosocial factors. Adequately powered factorial RCTs stratified by baseline Omega-3 Index, 25(OH)D, and vitamin K status, with prespecified mechanistic biomarkers and hard endpoints, are required.

Background: Keratoconus is a progressive corneal ectasia characterised by extracellular matrix (ECM) loss and an emerging inflammatory component, for which no disease-modifying molecular therapy exists. Exosomes derived from limbal and mesenchymal stem cells are an attractive cell-free therapeutic modality, but the cargo that should be delivered is undefined, and no curated limbal stem-cell (LSC) exosome cargo dataset currently exists. Methods: We reanalysed a public keratoconus corneal RNA-sequencing dataset (GEO: GSE77938; discovery and replication cohorts) with DESeq2, defined a replicated differentially expressed gene (DEG) set, and performed Gene Ontology, KEGG and Reactome enrichment. A high-confidence protein-protein interaction (PPI) network (STRING) identified hub genes. We integrated keratoconus disease-gene evidence (Open Targets Platform) and documented extracellular-vesicle cargo (ExoCarta, Vesiclepedia) and computed a transparent Cargo Prioritization Score (CPS) to nominate candidate LSC-exosome therapeutic cargo. Results: 1677 DEGs were detected in discovery (152 up, 1525 down) and 1380 were replicated. Enrichment was dominated by extracellular matrix organization; adaptive immune response; mononuclear cell differentiation. Network analysis nominated ECM and immune hub genes. The CPS prioritised COL1A1, FN1, COL4A1, COL3A1, COL5A1, MMP1 as leading restoration-cargo candidates, all documented as EV cargo and present in the mesenchymal stem-cell EV reference proteome. Conclusions: This fully reproducible, real-data framework provides a ranked, evidence-traceable shortlist of candidate LSC-exosome cargo for keratoconus and an explicit account of current data gaps to guide experimental validation.

Background: Population aging is driving a convergent rise in three disorders historically managed in isolation: cardiovascular disease, neurocognitive decline, and osteoporotic bone loss. Mechanistic data indicate that these systems are coupled through shared regulators of calcium trafficking, inflammation resolution, vascular integrity, and inflammaging. On this basis, a “Heart–Brain–Bone” axis has been proposed; it should be understood as an integrative conceptual framework that organizes evidence drawn from three separate studies, not as a validated physiological entity with agreed diagnostic criteria or demonstrated modifiability. Three lipid-soluble nutrients—long-chain omega-3 polyunsaturated fatty acids (EPA/DHA), vitamin D3 (cholecalciferol), and vitamin K2 (menaquinone-7 [MK-7])—act on overlapping nodes of this network. Methods: We performed a structured narrative review. PubMed/MEDLINE, Embase, the Cochrane Library, and Web of Science were searched from database inception to 25 June 2026 using predefined term blocks for each nutrient, each organ domain, and each candidate mechanism, and the search was updated on 7 August 2026. Records were screened against prespecified inclusion and exclusion criteria by two authors independently, with disagreements resolved by a third. The strength of evidence for each nutrient–organ relationship was graded with an explicitly defined four-level scheme ((−) to (+++)) applied separately to preclinical, observational, randomized and meta-analytic evidence. Results: Vitamin K2-dependent gamma-carboxylation of matrix Gla protein (MGP) and osteocalcin has been proposed to influence whether calcium is incorporated into the bone matrix or deposited in the arterial wall, offering a candidate mechanistic account of the “calcium paradox” associated with isolated vitamin D3 supplementation; EPA/DHA-derived specialized pro-resolving mediators may support resolution of endothelial and neuronal inflammation; and bone-, vascular- and brain-derived signals (osteocalcin, FGF23, the neurovascular unit) interconnect the three organs. These mechanisms are biologically plausible but remain insufficiently confirmed in humans. The clinical evidence is heterogeneous, formulation- and population-dependent, and comprises positive, neutral and null results: cardiovascular omega-3 trials are discordant (REDUCE-IT, which used icosapent ethyl [an EPA ethyl ester], positive; VITAL/STRENGTH/ASCEND null, predominantly in lower-risk or replete cohorts); cognitive trials are largely null or subgroup-dependent (MAPT, DO-HEALTH, VITAL); and MK-7 improves surrogate bone and calcification biomarkers and slowed coronary artery calcification in one recent randomized imaging trial (VitaK-CAC), whereas combined MK-7 plus vitamin D3 did not slow aortic valve or coronary calcification in AVADEC and MK-7 did not reduce bone loss in early menopausal women. Recognized safety signals include a dose-dependent increase in atrial fibrillation with high-dose omega-3, adverse skeletal effects of high-dose or bolus vitamin D, and clinically relevant interference of even low-dose MK-7 with vitamin K antagonist therapy. Conclusions: No adequately powered randomized trial has demonstrated that the combination of long-chain omega-3, vitamin D3 and MK-7 is superior to its individual components or to placebo for any clinical endpoint. The combined regimen is therefore mechanistically rational and hypothesis-generating rather than clinically established; benefit appears most plausible in individuals with elevated risk or demonstrable nutritional insufficiency, and least in replete, low-risk populations. Findings should be interpreted within a broader healthy-aging context that includes lifestyle and psychosocial factors. Adequately powered factorial randomized controlled trials stratified by baseline Omega-3 index, 25(OH)D and vitamin K status, with prespecified mechanistic biomarkers and hard endpoints, are required.

Background: Mesenchymal stromal cell (MSC)-derived extracellular vesicles (EVs) are under investigation as cell-free therapeutic platforms for neurodegeneration and age-related disease, and their biological effects depend partly on cargo composition. Existing MSC-EV cargo studies rarely integrate disease-gene evidence, aging databases, publication-level support and translational annotation. Methods: Using public data only, we assembled documented human MSC-EV cargo from ExoCarta and Vesiclepedia, deduplicated to independent source publications, and retained proteins with support from at least two publications. Each protein was integrated across independent-publication support, neurodegeneration disease-gene evidence (Open Targets), aging/senescence membership (CellAge, GenAge) and protein-interaction centrality (STRING) into a four-component Cargo Prioritization Score (CPS), after a component-correlation analysis. Robustness was assessed by weighting, component and source-study analyses; enrichment used the MSC-EV cargo universe as background; and top candidates received a separate Computational Liability Annotation (LOEUF, oncology association, expression breadth) with structured directionality curation. Results: The reproducible universe comprised 815 proteins derived from 9 source publications; disease-breadth was removed for redundancy with the neurodegeneration score (Spearman ρ=0.93). The ranking was robust to weighting (equal-weight ρ≈0.99) and to removal of any single component (ρ=0.83–0.94), but the eligible universe and several disease-anchored candidates depended strongly on individual source publications: excluding one publication reduced the universe from 815 to 228 proteins. 43 proteins carried joint neurodegeneration and aging evidence, including VCP, PARK7, SQSTM1, SOD1 and APOE, whereas the highest ranks partly reflected abundant, broadly expressed or highly connected proteins (GAPDH, ACTB, ALDOA, EGFR). Conclusions: The framework integrates evidence to prioritize MSC-EV cargo and separately annotates translational directionality and liability. A high CPS does not establish beneficial delivery, and the annotation is not a safety assessment; the output is a translationally annotated, hypothesis-generating candidate set for product-specific experimental validation.