ObjectiveTo explore the antibacterial effect of ginkgolic acid (GA) modified into liposome nanoparticles (TLM@GA) against Helicobacter pylori and investigate the antibacterial mechanism involved to provide a lead drug for radical clinical treatment.MethodsA new complex that improved GA stability and bioavailability was constructed using liposome nanoparticles (TLM@GA). The construction of the complex was verified by Fourier transform infrared spectroscopy, scanning electron microscopy, Zeta potential, and particle size detection, and its physicochemical properties were comprehensively evaluated. An acute gastritis model was also established using C57BL/6 mice, in which the in vivo therapeutic effect of TLM@GA was evaluated by detecting the colonization amount of the clinical multi-resistant strain HPBS001 in the gastric mucosa. Additionally, the pathological inflammation of the gastric mucosa was observed with H&E staining, apoptosis of gastric mucosal cells was observed with TUNEL staining, and the expression of serum inflammatory factors was detected with enzyme-linked immunosorbent assay. The in vivo safety of TLM@GA was evaluated by detecting its effect on mouse body weight and the damage to the stomach, liver, spleen, and kidneys.ResultsThe TLM@GA complex had excellent stability and dispersibility. The minimum inhibitory concentration of GA against H. pylori strains was 16–32 μg/mL, and that of TLM@GA was 0.25–0.5 μg/mL. After GA was modified into TLM@GA, its antibacterial activity was 32–128 times higher. After treatment with TLM@GA (28 mg/kg), the colonization of H. pylori in the gastric mucosa was significantly reduced, which was better than that in the omeprazole and amoxicillin (dual-therapy) and GA groups. Apoptotic and inflammatory cells in the gastric mucosa were significantly reduced, indicating alleviated inflammation. After treatment, the expression levels of major inflammatory factors were significantly decreased. There was no significant change in body weight when mice were intragastrically administered 140 mg/kg TLM@GA for 1 week, and no obvious pathological damage was found in the stomach, liver, spleen, or kidneys.ConclusionTLM@GA had excellent stability, efficient release, and good loading capacity efficiency. It was significantly superior to GA in terms of in vitro antibacterial activity, had low toxicity, was not prone to drug resistance, and demonstrated high safety. TLM@GA demonstrated a good antibacterial effect in the in vivo acidic environment, effectively slowing the apoptosis of gastric mucosal cells and significantly reducing levels of inflammatory factors, alleviating the inflammatory response.

Alzheimer's disease (AD) involves progressive neurodegeneration, with abnormal receptor signaling and disrupted cell-cycle activity leading to neuronal loss. Here, we identify a previously unknown mechanism linking β-amyloid (Aβ) exposure to the nuclear translocation of the Insulin-like Growth Factor 1 Receptor (IGF1R) in differentiated SH-SY5Y neuronal cells. The differentiated cholinergic model induced by retinoic acid and BDNF expresses acetylcholinesterase (AChE) and indicates that under amyloidogenic stress, IGF1R may transition from homeostatic membrane and vesicular signaling to a nuclear-centric function. We show that prolonged Aβ treatment causes phosphorylation-dependent nuclear import of IGF1R, confirmed by confocal imaging and biochemical fractionation. IGF1R is conventionally located in the membrane and vesicular membranes; however, under amyloidogenic stress, we show here that it is imported to the nucleus and exerts transcriptional control. The buildup of nuclear IGF1R coincided with increased Cyclin D1 levels and redistribution of neurons into S and G₂ phases, indicating abnormal cell-cycle re-entry. Chromatin immunoprecipitation demonstrated increased IGF1R binding at the CCND1 and JUN promoters after Aβ exposure, suggesting a direct role in gene transcription. Pharmacological blockade of IGF1R phosphorylation by PPP or SUMOylation by Ginkgolic acid significantly reduced Cyclin D1 elevation, implying that both post-translational modifications are involved in receptor nuclear trafficking. Co-immunoprecipitation and confocal imaging identified Nucleophosmin (NPM1) as a putative IGF1R interacting partner, potentially contributing to its nuclear transport and stabilizing receptor-chromatin complexes. These results establish IGF1R as a signaling-transcription connector linking extracellular amyloid stress to nuclear gene regulation, providing a mechanistic explanation for faulty neuronal cell-cycle re-entry in AD. We suggest that abnormal IGF1R-NPM1 interactions contribute to receptor mislocalization and cell-cycle failure, highlighting new targets for therapeutic intervention aimed at receptor trafficking and neuroprotection in Alzheimer's disease.

Context Ginkgolic acid (GAC), one of the major active constituents of Ginkgo biloba L. (Ginkgoaceae) extract, has been reported as a potential anticancer agent. Objective To investigate the effect of GAC on the viability of human hepatocellular carcinoma (HCC) cells and to identify its primary target and underlying mechanism. Materials and methods Human HCC cell lines and an orthotopic HCC mouse model were employed. The mechanism of GAC was elucidated through integrated analyses of cell viability, RNA sequencing, and biomarkers. The primary target of GAC was identified by drug affinity responsive target stability (DARTS) coupled with LC-MS/MS and further validated using molecular docking (MD), cellular thermal shift assay (CETSA), and microscale thermophoresis (MST) assay. Mechanistic roles were confirmed using genetic approaches, including gene knockdown and the construction of mutant plasmids. A KFERQ reporter system was used to detect the activity of chaperone-mediated autophagy (CMA). Results GAC effectively inhibits the viability of HCC by triggering ferroptosis. HSPA8 was identified as the direct target of GAC. The binding of GAC with HSPA8 enhances its interaction with glutathione peroxidase 4 (GPX4), which leads to the degradation of GPX4 via CMA. This process depleted glutathione (GSH) and caused lipid peroxidation, finally inducing ferroptosis in HCC cells. Furthermore, GAC suppressed tumor growth in an orthotopic HCC model, increased lipid peroxidation, and GPX4 degradation in tumor tissues. Discussion and conclusions Our results revealed a novel mechanism by which GAC induces ferroptosis in HCC cells through direct targeting of HSPA8 and promoting CMA-dependent GPX4 degradation. These findings suggest GAC-mediated ferroptosis as a potential therapeutic strategy against HCC and expand the pharmacological application of CMA-targeted cancer therapies.

Ginkgolic acid ameliorates pressure overload-induced cardiac remodeling by inhibiting SAE1 and activating the NRF2/HMOX1 pathway.

Ginkgo biloba is a singular and relict gymnosperm indigenous to China. Its distinctive fleshy episperm is rich in unique metabolites, ginkgolic acids, which protect the developing seed from biotic stresses. The unique nature of the tissue and its metabolites has made it highly challenging to elucidate the molecular and cellular mechanisms governing ginkgolic acid biosynthesis and regulation. In this study, we performed the mass spectrometry imaging of G. biloba seed, revealing that ginkgolic acids primarily accumulate in the secretory cavities of the episperm. We constructed a single-cell expression atlas of the G. biloba episperm and identified seven cellular types: meristem cells, subepidermal cells, lignified cells, trancheid cells, parenchymal cells, secretory cavity cells, and epidermis cells. Based on the analysis of upregulated gene expression in secretory cavity cells, pseudotime analysis of cell differentiation, and gene expression trajectory analysis, we precisely identified the key enzyme-encoding genes highly associated with ginkgolic acid biosynthesis. This approach elucidated the cellular and molecular mechanisms underlying secretory cell differentiation, secretory cavity formation, and ginkgolic acid biosynthesis and accumulation in response to exogenous jasmonic acid induction. By constructing a molecular interaction network, it was determined that the GbWRKY35, encoded by Gb_25334, is the core transcription factor. We further identified the signaling proteins that interact with GbWRKY35, confirming its central positive regulatory role in ginkgolic acid biosynthesis. As a core transcription factor, GbWRKY35 regulates ginkgolic acid biosynthesis through stimulating the expression of GbAAE16. This study provides the first spatially resolved investigation into the molecular and cellular regulatory mechanisms of ginkgolic acid biosynthesis in the episperm under jasmonic acid induction.

The SUMOylation modification is closely linked to the progression of fibrotic diseases, yet its role in hepatic fibrosis associated with cystic echinococcosis (CE) remains unclear. This study aimed to investigate the function of SUMOylation in CE-related hepatic fibrosis and evaluate the anti-fibrotic effects and mechanisms of ginkgolic acid (GA) via regulation of the SUMOylation pathway. Peri-lesional (PL) and adjacent normal (AN) liver tissues from CE patients were collected to examine histopathology and SUMO pathway proteins. A CE-infected mouse model was established and treated with GA to assess cyst burden, serum TGF-β1 levels, hepatic fibrosis markers, and SUMO-related proteins. In vitro, macrophages and hepatic stellate cells (HSCs, LX-2 line) were stimulated with Echinococcus granulosus cyst fluid (EgCF) or TGF-β1 to evaluate GA's effects on macrophage polarization (CD206/CD86), HSC activation (α-SMA/PCNA), Smad4 SUMOylation, and nuclear translocation. Macrophage-HSC crosstalk was investigated via conditioned medium co-culture assays. Fibrosis was exacerbated in peri-lesional liver tissues of CE patients, accompanied by SUMO pathway activation. GA significantly alleviated hepatic fibrosis in CE mice and reversed SUMO pathway dysregulation. Mechanistically, GA inhibited EgCF-induced pro-fibrotic M2 macrophage polarization and blocked Smad4 SUMOylation and nuclear translocation by modulating SUMOylation. Furthermore, GA directly suppressed HSC activation and bidirectionally disrupted the pro-fibrotic crosstalk between macrophages and HSCs under EgCF stimulation, ultimately alleviating fibrosis. This study reveals the critical role of SUMOylation modification in CE-associated hepatic fibrosis and elucidates a novel anti-fibrotic mechanism whereby GA targets the SUMOylation-Smad4 axis to regulate the immune microenvironment.

Ginkgolic acid: a novel IL1R1 agonist facilitating megakaryocyte differentiation and platelet production via the SRC/MEK/ERK pathway.

Ginkgolic acid (GA) exhibits various biological activities, but its role in vascular restenosis remains unreported. GA (13:0) is a relatively abundant natural congener. This study aims to investigate and clarify the effects and mechanisms of GA (13:0) on vascular smooth muscle cell (VSMC) proliferation and migration in vitro, as well as on balloon injury-induced vascular restenosis in rats. The results showed that GA (13:0) significantly inhibited VSMC proliferation, migration, and intimal thickening both in vitro and in vivo. Moreover, GA (13:0) reduced the expression of cyclin D1, cyclin E1, CDK2, and CDK4, as well as cyclin D1-CDK4 and cyclin E1-CDK2 binding, leading to G0/G1 arrest. Additionally, GA (13:0) suppressed vimentin expression and actin cytoskeleton polymerization and altered F-actin morphology. Comparative proteomics identified tectonic family member 1 (TCTN1) as a potential molecular target of GA (13:0). GA (13:0) reduced TCTN1 expression both in vitro and in vivo. Crucially, TCTN1 overexpression notably reversed the inhibitory effects of GA (13:0) on VSMC proliferation, migration, intimal thickening, expression and binding of cell cycle-related proteins, and vimentin expression. Concurrently, TCTN1 overexpression also reversed GA (13:0)-induced F-actin depolymerization and rearrangement and G0/G1 arrest. GA (13:0) significantly inhibited TCTN1 co-localization with vimentin and actin in vitro and in vivo. Furthermore, we found that CCCTC binding factor (CTCF) binds to the 162–176 site of the TCTN1 promoter to regulate TCTN1 transcription, and CTCF knockout significantly down-regulated TCTN1 protein levels. This study reveals that GA (13:0) inhibits TCTN1 transcription and expression, hindering G1/S transition, vimentin expression, and F-actin rearrangement, thereby suppressing vascular restenosis.

Abstract Arsenic trioxide (ATO) therapy is limited by its severe vascular toxicity, which manifests as endothelial injury through mechanisms that remain poorly defined. This study identifies the SUMOylation of the Transient Receptor Potential Melastatin 4 (TRPM4) channel as a pivotal pathological driver of this process. We demonstrate that the SUMOylation inhibitor ginkgolic acid (GA) confers protection by attenuating ATO‐induced TRPM4 overactivation, thereby rescuing endothelial cells from ionic dysregulation (Ca 2 ⁺, Na⁺), oxidative stress, membrane depolarization, and apoptosis. Critically, we identified lysine 618 (K618) as the key SUMOylation site on TRPM4. Expression of a SUMOylation‐deficient TRPM4 mutant (K618R) phenocopied the protective effects of GA, significantly mitigating ATO‐induced endothelial dysfunction. This confirms that the detrimental effects of TRPM4 are strictly dependent on its SUMOylation at this residue. The protective efficacy of GA was further validated ex vivo, where it restored endothelium‐dependent vasodilation in rat mesenteric arteries impaired by ATO. Complementing these findings, Raman microimaging provided unique, label‐free insights into the biochemical alterations induced by ATO and their reversal by GA at the single‐cell level. Our findings unveil TRPM4‐SUMOylation at K618 as a fundamental mechanism underlying ATO‐induced endothelial injury. These results position the inhibition of this specific post‐translational modification as a promising therapeutic strategy for managing the vascular toxicity associated with ATO chemotherapy.

This study established an efficient in vitro regeneration system using stem nodes from root collar suckers as explants. Subsequently, regenerated shoots were used to establish an in vitro medicinal production protocol that achieved ginkgolic acid production. The self-developed Ginkgo biloba medium (GBM), first reported in this study, was pivotal to system establishment. The plantlet propagation system showed that the bases of stem nodes dipped in GBM with 2 mg·L−1 6-benzyladenine (BA) and 0.2 mg·L−1 1-naphthaleneacetic acid (NAA) achieved near-complete axillary bud induction (99.56%). Adventitious shoot induction reached 82.22% (3.5 shoots/explant) using GBM with 0.2 mg·L−1 BA, 0.02 mg·L−1 kinetin (Kin) and 0.2 g·L−1 proline (Pro). Maximum adventitious shoot elongation (92.22%, average 3.35 cm) was observed on GBM containing 0.1 mg·L−1 zeatin (ZT) and 0.01 mg·L−1 BA. After 3-week preculture with 15 mg·L−1 phloroglucinol (PG), treatment with 0.6 mg·L−1 indole-3-butyric acid (IBA) and 0.2% activated carbon (AC) yielded 96.67% rooting (6.19 roots/explant) and 85% acclimatization survival. For medicinal resource production, bud cluster induction at 94.44% (20.89 buds/explant) on GBM with 1 mg·L−1 BA, 0.03 mg·L−1 Kin, and 0.2 g·L−1 Pro. Leaf organs in GBM with 0.3 mg·L−1 BA, 0.01 mg·L−1 Kin, 0.01 mg·L−1 IBA, 0.3 g·L−1 Pro, and 0.01 mg·L−1 glutamine (Gln) accumulated 20.64 g fresh weight and 41.910 mg·g−1 DW ginkgolic acids, representing a 4.93-fold increase over mother plants. This system enables large-scale Ginkgo biloba L. propagation and provides an in vitro strategy for producing medicinal compounds in endangered plants.

Background This study investigates Ginkgolic acid, a naturally occurring phenolic compound from Ginkgo biloba, for its potential to inhibit SARS-CoV-2 by targeting the RBD–ACE2 interface through a combination of computational and in vitro methodologies. Methods In silico molecular docking and 500 ns molecular dynamics (MD) simulations were employed to examine the binding conformation and stability of Ginkgolic acid at the RBD–ACE2 interface. In vitro studies included cytotoxicity profiling, plaque reduction assays, qRT-PCR, luciferase-based viral entry quantification, and membrane fusion inhibition in Vero E6 and ACE2-expressing HEK293T cells. Results Ginkgolic acid demonstrated dose-dependent antiviral activity in Vero E6 cells with half-maximal inhibitory concentration (IC50) values ranging from 0.025 to 0.102 µM. MD simulations revealed its strong binding affinity at the RBD–ACE2 interface, particularly through interactions with conserved residues His34 and Asp38, leading to increased inter-residue distances and destabilization of the complex. In vitro assays confirmed significant virucidal activity, reduced viral entry, and inhibition of spike-mediated membrane fusion. Despite limited intracellular uptake, its antiviral efficacy appears to be predominantly extracellular. Conclusion Ginkgolic acid acts as an allosteric modulator and fusion inhibitor by targeting conserved RBD–ACE2 interface residues, disrupting viral attachment and fusion. Its dual mechanism—direct virucidal effect and host-cell entry inhibition—presents a promising scaffold for future antiviral development. These findings warrant further preclinical validation to assess its efficacy across SARS-CoV-2 variants.

ABSTRACTCentrins, Ca2+‐binding proteins conserved in eukaryotes, are the key components of the microtubule‐organizing center. Giardia lamblia possesses two centrins (GL50803_6744: centrin 1; GL50803_104685: centrin 2) localized in the basal bodies during cell division. G. lamblia centrin 2 (Glcent2) is also found in the nuclei of interphase Giardia, with its N‐terminal half being necessary for this localization. Morpholino‐mediated knockdown of Glcents resulted in abnormal nuclear positioning and cytokinesis, causing cell malformations, including ventral discs and flagella defects. Small ubiquitin‐like modifier (SUMO)ylation is a posttranslational modification, which modulates several cellular processes. Here, we demonstrated that Glcents are substrates of SUMO through in vitro SUMOylation and immunoprecipitation experiments. Additionally, treatment of Giardia with ginkgolic acid, which inhibits the E1 enzyme of the SUMO pathway, and CRISPRi‐mediated inhibition of G. lamblia Ubc9, the E2 conjugation enzyme involved in SUMOylation, resulted in defects in the localization of Glcents. Blocking SUMOylation resulted in the arrest of Giardia cells and conformational changes, including alterations in the ventral disc shape, posterolateral flanges, and peripheral vesicles. Taken together, we demonstrated that Glcents function in Giardia cell cycle progression and morphogenesis, with the activity of both Glcents being modulated by SUMOylation.

Когнитивные расстройства являются одной из самых актуальных проблем в области неврологии. Эта категория заболеваний включает в себя широкий спектр нарушений, касающихся памяти, внимания, восприятия и других познавательных функций. Особое внимание следует уделять выявлению этих расстройств на ранних, додементных стадиях. Раннее обнаружение когнитивных нарушений имеет критическое значение, поскольку на этой стадии возможна реализация медицинских мероприятий, способствующих замедлению или даже приостановке прогрессирования когнитивного снижения. Важно отметить, что многие пациенты не осознают свои когнитивные проблемы, а их близкие часто воспринимают изменения в поведении как естественные возрастные изменения. В обзоре приводятся данные о влиянии на развитие когнитивного дефицита таких факторов, как возраст, сердечно-сосудистые заболевания, эмоциональные нарушения и т.д. Наличие двух или более таких факторов риска, как правило, увеличивает вероятность развития болезни Альцгеймера. Рассматриваются возможности медикаментозной профилактики нарастания когнитивного дефицита при начальных формах когнитивного снижения наряду с немедикаментозными методами коррекции, которые в настоящее время являются основными при ведении подобных пациентов. Обсуждается возможность применения обладающего многофакторным механизмом действия экстракта гинкго билоба, который отвечает регламентированному содержанию активных ингредиентов (флавоновых гликозидов, терпенлактонов и гинкголиевой кислоты), и препаратов, направленных на восстановление или модуляцию холинергической активности. Приводятся данные исследования комбинированной формулы продукта Холин Стронг®, которая представляет собой фиксированную комбинацию холина битартрата и экстракта гинкго билоба. Исследование показало потенциальное фармакокинетическое преимущество данного продукта и обосновывает необходимость дальнейших исследований для уточнения механизмов его действия. Cognitive disorders are one of the most pressing problems in the field of neurology. This category of diseases includes a wide range of disorders related to memory, attention, perception and other cognitive functions. Special attention should be paid to the detection of these disorders in the early, pre-stage stages. Early detection of cognitive impairment is of critical importance, since at this stage it is possible to implement medical measures that help slow down or even stop the progression of cognitive decline. It is important to note that many patients are unaware of their cognitive problems, and their loved ones often perceive behavioral changes as natural age-related changes. The review provides data on the influence of such factors as age, cardiovascular diseases, emotional disorders, etc. On the development of cognitive deficits. The presence of two or more such risk factors, as a rule, increases the likelihood of developing Alzheimer’s disease. The possibilities of drug prevention of the increase in cognitive deficits in the initial forms of cognitive decline are considered along with non-drug correction methods, which are currently the main ones in the management of such patients. The possibility of using ginkgo biloba extract with a multifactorial mechanism of action, which meets the regulated content of active ingredients (flavone glycosides, terpenactones and ginkgolic acid), and drugs aimed at restoration or modulation of cholinergic activity. The data from the study of the combined formula of the Choline Strong, which is a fixed combination of choline bitartrate and ginkgo biloba extract, are presented. The study showed the potential pharmacokinetic advantage of this product and justifies the need for further research to clarify the mechanisms of its action.

A "cyclodextrin-salicylic acid-chitosan" bifunctional monomer magnetic hydrophilic imprinted sandwich gel for targeted adsorption and slow release of ginkgolic acid.

Vascular endothelial cells are the predominant cell type in the cardiovascular system, and their dysfunction and death following hypoxic injury contribute to vascular lesions, playing an essential role in cardiovascular disease. Despite its importance, the mechanisms underlying vascular endothelial cell injury under hypoxia and potential therapeutic interventions remain poorly understood. Here, we constructed both an in vivo hypoxia model in C57BL/6 mice and an in vitro hypoxia model in HUVEC cells. Our findings demonstrated that hypoxia induces necroptosis in vascular endothelial cells and exacerbates inflammatory injury in vivo and in vitro, as evidenced by immunofluorescence and western blot. We identified FADD as a critical regulator of hypoxia-mediated necroptosis, with FADD knockdown significantly reversing hypoxia-induced necroptosis. Mechanistically, hypoxia affected protein conformation through SUMOylation of FADD and competitively inhibited its ubiquitination, leading to an increase in protein half-life and protein level of FADD. Furthermore, SUMOylation increased the interaction between FADD and RIPK1 and induced the formation of the FADD-RIPK1-RIPK3 complex, thereby promoting necroptosis in vascular endothelial cells. The SUMOylation inhibitor ginkgolic acid (GA) notably reduced hypoxia-induced vascular endothelial injury and inflammatory responses in male mice. Taken together, our research has uncovered a new process by which SUMOylation of FADD regulates hypoxia-induced necroptosis in endothelial cells, providing potential therapeutic targets for hypoxia-related cardiovascular diseases.

Ginkgolic acid inhibits CD8+ T cell activation and induces ferroptosis by lactate dehydrogenase A to exert immunosuppressive effect

Ginkgolic acid inhibits Ebola virus transcription and replication by disrupting the interaction between nucleoprotein and VP30 protein.

Aging is a major risk factor for Alzheimer's disease (AD). With the prevalence of AD increased, a mechanistic linkage between aging and the pathogenesis of AD needs to be further addressed. Here, we report that a small ubiquitin-related modifier (SUMO) modification of p53 is implicated in the process which remarkably increased in AD patient's brain. Mechanistically, SUMOylation of p53 at K386 residue causes the dissociation of SET/p53 complex, thus releasing SET into the cytoplasm, SET further interacts with cytoplasmic PP2A and inhibits its activity, resulting in tau hyperphosphorylation in neurons. In addition, SUMOylation of p53 promotes the p53 Ser15 phosphorylation that mediates neuronal senescence. Notably, p53 SUMOylation contributes to synaptic damage and cognitive defects in AD model mice. We also demonstrate that the SUMOylation inhibiter, Ginkgolic acid, recovering several senescent phenotypes drove by p53 SUMOylation in primary neurons. These findings suggest a previously undiscovered etiopathogenic relationship between aging and AD that is linked to p53 SUMOylation and the potential of SUMOylated p53-based therapeutics for neurodegeneration such as Alzheimer's disease.

Fibrosis is the outcome of any abnormal tissue repair process that results in normal tissue replacement with scar tissue, leading to persistent tissue damage and cellular injury. During the process of fibrosis, many cytokines and chemokines are involved, and their activities are controlled by post-translational modifications, especially SUMOylation and NEDDylation. Both these modifications entail a three-step process of activation, conjugation, and ligation that involves three kinds of enzymes, namely, E1 activating, E2 conjugating, and E3 ligase enzymes. SUMOylation participates in organ fibrosis by modulating FXR, PML, TGF-β receptor I, Sirt3, HIF-1α, and Sirt1, while NEDDylation influences organ fibrosis by regulating cullin3, NIK, SRSF3, and UBE2M. Further investigations exhibit the therapeutic potentials of SUMOylation/NEDDylation activators and inhibitors against organ fibrosis, especially ginkgolic acid in SUMOylation and MLN4924 in NEDDylation. These results demonstrate the therapeutic effects of SUMOylation and NEDDylation against organ fibrosis and highlight their activators as well as inhibitors as potential candidates. In the future, deeper investigations of SUMOylation and NEDDylation are needed to identify novel substrates against organ fibrosis; moreover, clinical investigations are needed to determine the therapeutic effects of their activators and inhibitors that can benefit patients. This review highlights that SUMOylation and NEDDylation function as potential therapeutic targets for organ fibrosis.

BackgroundCancer-associated fibroblasts (CAFs) are the most abundant stromal cellular component in the tumor microenvironment (TME). CAFs contribute to tumorigenesis and have been proposed as targets for anticancer therapies. Similarly, dysregulation of SUMO machinery components can disrupt the balance of SUMOylation, contributing to tumorigenesis and drug resistance in various cancers, including breast cancer. We explored the role of SUMOylation in breast CAFs and evaluated its potential as a therapeutic strategy in breast cancer.MethodsWe used pharmacological and genetic approaches to analyse the functional crosstalk between breast tumor cells and CAFs. We treated breast CAFs with the SUMO1 inhibitor ginkgolic acid (GA) at two different concentrations and conditioned media was used to analyse the proliferation, migration, and invasion of breast cancer cells from different molecular subtypes. Additionally, we performed quantitative proteomics (SILAC) to study the differential signalling pathways expressed in CAFs treated with low or high concentrations of GA. We confirmed these results both in vitro and in vivo. Moreover, we used samples from metastatic breast cancer patients to evaluate the use of GA as a therapeutic strategy.ResultsInhibition of SUMOylation with ginkgolic acid (GA) induces death in breast cancer cells but does not affect the viability of CAFs, indicating that CAFs are resistant to this therapy. While CAF viability is unaffected, CAF-conditioned media (CM) is altered by GA, impacting tumor cell behaviour in different ways depending on the overall degree to which SUMO1-SUMOylated proteins are dysregulated. Breast cancer cell lines exhibited a concentration-dependent response to conditioned media (CM) from CAFs. At a low concentration of GA (10 µM), there was an increase in proliferation, migration and invasion of breast cancer cells. However, at a higher concentration of GA (30 µM), these processes were inhibited. Similarly, analysis of tumor development revealed that at 10 µM of GA, the tumors were heavier and there was a greater degree of metastasis compared to the tumors treated with the higher concentration of GA (30 µM). Moreover, some of these effects could be explained by an alteration in the activity of the GTPase Rac1 and the activation of the AKT signalling pathway. The results obtained using SILAC suggest that different concentrations of GA affected cellular processes differentially, possibly influencing the secretome of CAFs. Treatment of metastatic breast cancer with GA demonstrated the use of SUMOylation inhibition as an alternative therapeutic strategy.ConclusionThe study highlights the importance of SUMOylation in the tumor microenvironment, specifically in cancer-associated fibroblasts (CAFs). Targeting SUMOylation in CAFs affects their signalling pathways and secretome in a concentration-dependent manner, regulating the protumorigenic properties of CAFs.