Abstract BHLHE40, a member of the basic helix‐loop‐helix transcription factor family, has been reported to play an important role in inflammatory diseases. However, the regulation and function of BHLHE40 in Helicobacter pylori ( H pylori )‐associated gastritis is unknown. We observed that gastric BHLHE40 was significantly elevated in patients and mice with H pylori infection. Then, we demonstrate that H pylori‐ infected GECs express BHLHE40 via cagA ‐ERK pathway. BHLHE40 translocates to cell nucleus, and then binds to cagA protein ‐ activated p‐STAT3 (Tyr705). The complex increases chemotactic factor CXCL12 expression (production). Release of CXCL12 from GECs fosters CD4 + T cell infiltration in the gastric mucosa. Our results identify the cagA ‐BHLHE40‐CXCL12 axis that contributes to inflammatory response in gastric mucosa during H pylori infection.
Triple-negative breast cancer (TNBC) cells have not been usefully classified, and no targeted therapeutic plans are currently available, resulting in a high recurrence rate and metastasis potential. In this research, CD24high cells accounted for the vast majority of TNBC cells, and they were insensitive to Taxol but sensitive to ferroptosis agonists and effectively escaped phagocytosis by tumor-associated macrophages. Furthermore, the NF2–YAP signaling axis modulated the expression of ferroptosis suppressor protein 1 (FSP1) and CD24 in CD24high cells, with subsequent ferroptotic regulation and macrophage phagocytosis. In addition, a precision targeted therapy system was designed based on the pH level and glutathione response, and it can be effectively used to target CD24high cells to induce lysosomal escape and drug burst release through CO2 production, resulting in enhanced ferroptosis and macrophage phagocytosis through FSP1 and CD24 inhibition mediated by the NF2–YAP signaling axis. This system achieved dual antitumor effects, ultimately promoting cell death and thus inhibiting TNBC tumor growth, with some tumors even disappearing. The composite nanoprecision treatment system reported in this paper is a potential strategic tool for future use in the treatment of TNBC.
<div>Abstract<p>Actin cytoskeleton dynamic rearrangement is required for tumor cell metastasis and is a key characteristic of <i>Helicobacter pylori</i> (<i>H. pylori</i>)-infected host cells. Actin cytoskeleton modulation is coordinated by multiple actin-binding proteins (ABP). Through Kyoto encyclopedia of gene and genomes database, GEPIA website, and real-time PCR data, we found that <i>H. pylori</i> infection significantly induced L-plastin, a key ABP, in gastric cancer cells. We further explored the regulation and function of L-plastin in <i>H. pylori</i>–associated gastric cancer and found that, mechanistically, <i>H. pylori</i> infection induced gastric cancer cells to express L-plastin via <i>cagA</i>-activated ERK signaling pathway to mediate SP1 binding to L-plastin promoter. Moreover, this increased L-plastin promoted gastric cancer cell proliferation and migration <i>in vitro</i> and facilitated the growth and metastasis of gastric cancer <i>in vivo</i>. Finally, we detected the expression pattern of L-plastin in gastric cancer tissues, and found that L-plastin was increased in gastric cancer tissues and that this increase of L-plastin positively correlated with <i>cagA</i><sup>+</sup> <i>H. pylori</i> infection status. Overall, our results elucidate a novel mechanism of L-plastin expression induced by <i>H. pylori</i>, and a new function of L-plastin–facilitated growth and metastasis of gastric cancer, and thereby implicating L-plastin as a potential therapeutic target against gastric cancer.</p>Implications:<p>Our results elucidate a novel mechanism of L-plastin expression induced by <i>H. pylori</i> in gastric cancer, and a new function of L-plastin–facilitated gastric cancer growth and metastasis, implicating L-plastin as a potential therapeutic target against gastric cancer.</p></div>
<p>Figure S1. NK cell and T cell numbers in the peripheral blood, non-tumor and tumor tissues of GC patients. Figure S2. Expression of activating and inhibitory receptors on NK cells in the peripheral blood, non-tumor and tumor tissues of GC patients. Figure S3. Expression of CD56 and CD16 on NK cells in the peripheral blood, non-tumor and tumor tissues of GC patients. Figure S4. The expression of HLA-DR on tissue-associated monocytes/macrophages in GC patients. Figure S5. The expression of CD48, PD-L1 and PD-L2 on tissue-associated monocytes/macrophages in GC patients. Figure S6. Co-expression of CD68 and TGF-beta1 in tumors of GC patients. Figure S7. Surface expression levels of TGF-beta1 on tumor-associated monocytes/macrophages in GC patients. Table S1. Clinical characteristics of 65 GC patients. Table S2. Fluorochrome-conjugated antibodies used in flow cytometry. Table S3. Univariate and multivariate analyses of factors associated with survival.</p>
Abstract Background: Helicobacter pylori (H. pylori) is a human pathogen that infects nearly half of the world’s population, however, the persistent colonization of H. pylori in gastric mucosa remains poorly understood. Nowadays it is believed that impairment of host defense of gastric epithelium induced by H. pylori plays key roles in H. pylori-associated pathology. The nuclear receptor Rev-erbα represents a powerful transcriptional repressor involved in host immunity. However, the regulation, function, and clinical relevance of Rev-erbα in H. pylori infection are presently unknown. Here we demonstrated a pro-colonization role of Rev-erbα in H. pylori infection. Results: Rev-erbα was increased in gastric mucosa of H. pylori-infected patients and mice. H. pylori induced gastric epithelial cells (GECs) to express Rev-erbα via the phosphorylated cagA that activated extracellular signal-regulated kinase (ERK) signaling pathway to mediate transcription factor nuclear factor kappa-B (NF-κB) directly binding to Rev-erbα promoter. Human gastric Rev-erbα expression correlated with H. pylori colonization, and mouse Rev-erbα from non-bone marrow-derived cells promoted gastric H. pylori burden. Importantly, H. pylori colonization was attenuated in Rev-erbα-/- mice and the mice with in vivo pharmacological inhibition of Rev-erbα. Mechanistically, Rev-erbα in GECs not only directly suppressed Reg3b and β-defensin-1 expression via binding to Reg3b and β-defensin-1 promoter respectively, which resulted in impaired bactericidal effects against H. pylori of these antibacterial proteins in vitro and in vivo; but also directly inhibited chemokine CCL21 expression via binding to CCL21 promoter, which led to decreased gastric influx of CD45+CD11c-Ly6G-CD11b+CD68- myeloid cells by CCL21-CCR7-dependent migration and, as a direct consequence, reduced bacterial clearing capacity of H. pylori-specific T helper type 1 (Th1) cell response. Conclusions: Overall, this study identifies a model involving Rev-erbα, which collectively ensures gastric bacterial persistence by suppressing host gene expression required for local innate and adaptive defense against H. pylori, and also highlight a pathological role and an immunosuppressive mechanism of Rev-erbα in persistent H. pylori infection.
Background & AimsRev-erbα represents a powerful transcriptional repressor involved in immunity. However, the regulation, function, and clinical relevance of Rev-erbα in Helicobacter pylori infection are presently unknown.MethodsRev-erbα was examined in gastric samples from H pylori-infected patients and mice. Gastric epithelial cells (GECs) were isolated and infected with H pylori for Rev-erbα regulation assays. Gastric tissues from Rev-erbα–/– and wild-type (littermate control) mice or these mice adoptively transferred with CD4+ T cells from IFN-γ–/– and wild-type mice, bone marrow chimera mice and mice with in vivo pharmacological activation or inhibition of Rev-erbα were examined for bacteria colonization. GECs, CD45+CD11c–Ly6G–CD11b+CD68– myeloid cells and CD4+ T cells were isolated, stimulated and/or cultured for Rev-erbα function assays.ResultsRev-erbα was increased in gastric mucosa of H pylori-infected patients and mice. H pylori induced GECs to express Rev-erbα via the phosphorylated cagA that activated ERK signaling pathway to mediate NF-κB directly binding to Rev-erbα promoter, which resulted in increased bacteria colonization within gastric mucosa. Mechanistically, Rev-erbα in GECs not only directly suppressed Reg3b and β-defensin-1 expression, which resulted in impaired bactericidal effects against H pylori of these antibacterial proteins in vitro and in vivo; but also directly inhibited chemokine CCL21 expression, which led to decreased gastric influx of CD45+CD11c–Ly6G–CD11b+CD68– myeloid cells by CCL21-CCR7-dependent migration and, as a direct consequence, reduced bacterial clearing capacity of H pylori-specific Th1 cell response.ConclusionsOverall, this study identifies a model involving Rev-erbα, which collectively ensures gastric bacterial persistence by suppressing host gene expression required for local innate and adaptive defense against H pylori. Rev-erbα represents a powerful transcriptional repressor involved in immunity. However, the regulation, function, and clinical relevance of Rev-erbα in Helicobacter pylori infection are presently unknown. Rev-erbα was examined in gastric samples from H pylori-infected patients and mice. Gastric epithelial cells (GECs) were isolated and infected with H pylori for Rev-erbα regulation assays. Gastric tissues from Rev-erbα–/– and wild-type (littermate control) mice or these mice adoptively transferred with CD4+ T cells from IFN-γ–/– and wild-type mice, bone marrow chimera mice and mice with in vivo pharmacological activation or inhibition of Rev-erbα were examined for bacteria colonization. GECs, CD45+CD11c–Ly6G–CD11b+CD68– myeloid cells and CD4+ T cells were isolated, stimulated and/or cultured for Rev-erbα function assays. Rev-erbα was increased in gastric mucosa of H pylori-infected patients and mice. H pylori induced GECs to express Rev-erbα via the phosphorylated cagA that activated ERK signaling pathway to mediate NF-κB directly binding to Rev-erbα promoter, which resulted in increased bacteria colonization within gastric mucosa. Mechanistically, Rev-erbα in GECs not only directly suppressed Reg3b and β-defensin-1 expression, which resulted in impaired bactericidal effects against H pylori of these antibacterial proteins in vitro and in vivo; but also directly inhibited chemokine CCL21 expression, which led to decreased gastric influx of CD45+CD11c–Ly6G–CD11b+CD68– myeloid cells by CCL21-CCR7-dependent migration and, as a direct consequence, reduced bacterial clearing capacity of H pylori-specific Th1 cell response. Overall, this study identifies a model involving Rev-erbα, which collectively ensures gastric bacterial persistence by suppressing host gene expression required for local innate and adaptive defense against H pylori.
Background: Adrenomedullin (ADM) is a multifunctional peptide that is expressed by many surface epithelial cells, but its relevance to Helicobacter pylori (H. pylori)-induced gastritis is unknown.Methods: The expression of ADM was detected and localized in H. pylori infected patients and mice gastric. The signaling pathway of ADM expression was detected. The differentiation and proliferation of T cells was detected after stimulated with ADM. IL-12 was detected in macrophages after stimulated with ADM.Findings: Gastric ADM expression was elevated in gastric mucosa of H. pylori-infected patients and mice. In H. pylori-infected human gastric mucosa, ADM expression was positively correlated with the degree of gastritis, accordingly, blockade of ADM resulted in decreased inflammation within the gastric mucosa of H. pylori-infected mice. During H. pylori infection, ADM production was promoted via PI3K-AKT signaling pathway activation by gastric epithelial cells in a cagA-dependent manner, and resulted in increased inflammation within the gastric mucosa. This inflammation was characterized by the increased IFN-γ-producing T cells, whose differentiation was induced via the phosphorylation of AKT and STAT3 by ADM derived from gastric epithelial cells. ADM also induced macrophages to produce IL-12, which promoted the IFN-γ-producing T-cell responses, thereby contributing to the development of H. pylori associated gastritis. Accordingly, blockade of IFN-γ or knockout of IFN-γ decreased inflammation within the gastric mucosa of H. pylori-infected mice.Interpretation: This study identifies a novel regulatory network involving H. pylori, gastric epithelial cells, ADM, macrophages, T cells, and IFN-γ, which collectively exert a pro-inflammatory effect within the gastric microenvironment.Funding Statement: This work was funded by National Key Research and Development Program of China (grant number: 2016YFC1302200) and National Natural Science Foundation of China (grant number: 81870394).Declaration of Interests: The authors declare that they have no conflict of interest.Ethics Approval Statement: The study was approved by the Ethics Committee of XinQiao Hospital and Southwest Hospital of Third Military Medical University. All human subjects were adult, and the written informed consent was obtained from each subject. All animal experiments were performed in strict accordance with the Guide for the Care and Use of Laboratory Animals issued by the Ministry of Science and Technology of the People's Republic of China. All breeding and experiments were undertaken with review and approval from the Animal Ethical and Experimental Committee of Third Military Medical University.
<p>S1. The GEPIA website (based on TCGA data) revealed that 48 ABPs were significantly up-regulated in GC tissues. S2. H. pylori induces L-plastin expression in GC cells. S3. Direct contact is required for H. pylori induced L-plastin expression in GC cells. S4. The vacA was not involved in the induction of L-plastin. S5. Decreased expression of L-plastin in presence of U0126 during H. pylori infection. S6. L-plastin expression is increased in GC tissues.</p>
Regulated in development and DNA damage responses-1 (REDD1) is a conserved and ubiquitous protein, which is induced in response to multiple stimuli. However, the regulation, function and clinical relevance of REDD1 in Helicobacter pylori-associated gastritis are presently unknown.
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