Project description:WXT significantly reduced ectopic lesion volume, alleviated peritoneal adhesions, and decreased the proportion of M2 macrophages in both lesion tissues and peritoneal fluid, along with reduced lactate accumulation and suppressed M2-associated cytokine secretion. Single-cell transcriptomic analysis revealed enhanced glycolytic activity in eESCs and a positive correlation between M2 macrophage polarization and lactate-sensing gene signatures. WXT-containing serum inhibited eESC-derived lactate production, reduced H3K9la levels in macrophages, and blocked lactate-induced M2 polarization. Through UPLC-Q-TOF-MS/MS and molecular docking screening, we identified tormentic acid as a key absorbed component with strong predicted binding to both GLUT1 (a key glycolytic transporter) and p300 (a key regulator of histone lactylation). Tormentic acid reproduced the effects of WXT-containing serum, suppressing lactate production, reducing H3K9la levels, and attenuating M2 polarization. CUT&Tag and ChIP-qPCR further revealed that tormentic acid reduced H3K9la enrichment at the WNT2B promoter and suppressed its expression, defining a lactate-H3K9la-WNT2B axis through which tormentic acid ameliorates endometriosis.
Project description:Cancer-augmented lactogenesis has been described by the Warburg effect, and is associated with several major hallmarks of neoplasia. However, the non-metabolic functions of elevated lactate in physiology and disease remain unknown. Here we report histone lysine lactylation as a new type of epigenetic mechanism and as a functional destination for lactate. Histone lactylation is induced under glycolytic conditions such as hypoxia and M1 macrophage polarization. In the late phase of M1 macrophage polarization, increases in histone lactylation but not acetylation mark M2-like genes for activation. Our findings suggest a feedback mechanism of the innate immune system to switch from proinflammation to resolution through histone Kla-associated gene expression. This mechanism is implemented by the coopted function of lactate and histone lactylation in metabolism and epigenetics. Together, our study opens a new avenue for understanding function of lactate and glycolysis underlined diverse pathophysiological conditions.
Project description:In response to microenvironmental signals macrophages undergo different activation, indicated as classic/M1 and alternative/M2 polarization. C-Myc transcription factor could be an essential player in M2 polarization. Functional relevance of c-Myc in M2 macrophage biology is investigated by evaluating the effect of 100-58F4, on the transcriptional profile induced on human macrophages by IL-4. Human monocytes were obtained from normal donor buffy coats by two-step gradient centrifugation using Ficoll (Biochrom) and Percoll (Amersham). Non-adherent cells were discarded, and the purified monocytes were incubated for 7 days in RPMI 1640 (Biochom) supplemented with 10% FCS (HyClone) and 100 ng/mL M-CSF to obtain resting macrophages. Macrophage polarization was obtained by removing the culture medium and culturing cells in RPMI 1640 supplemented with 10% FCS and 100 ng/mL LPS plus 20 ng/mL IFN-gamma (M1 polarization) or 20 ng/mL IL-4 (M2 polarization) for 24 h. When needed, chemical inhibitors were added with IL-4.
Project description:Macrophage recruitment into tumors is correlated with poor outcomes in cancer, which correlate with STAT6-dependent M2 macrophage polarization and wound healing-type responses. Using penetrant, genetic models of neuroblastoma, we found macrophage recruitment was protective against tumor formation while disabling STAT6-mediated M2 polarization had no effect on tumorigenesis, progression or expression of key immunosuppressive pathways. Thus while macrophages are drivers of cancer in this model, non-classical M2 STAT6-independent pathways are plausible targets for cancer therapy.
Project description:In response to microenvironmental signals macrophages undergo different activation, indicated as classic/M1 and alternative/M2 polarization. C-Myc transcription factor could be an essential player in M2 polarization. Functional relevance of c-Myc in M2 macrophage biology is investigated by evaluating the effect of 100-58F4, on the transcriptional profile induced on human macrophages by IL-4.
Project description:Macrophages polarize towards different subpopulations with distinct and partly antagonistic functions in various diseases. IFNγ/LPS-polarized M1-type macrophages can have antiangiogenic activity, whereas IL-4-induced M2-type macrophages can be proangiogenic and profibrotic. Therapeutic strategies to inhibit M2-type polarization while promoting M1-type polarization could serve to inhibit pathological angiogenesis and fibrosis. Here, by combining global quantitative time-course proteomics and phosphoproteomics with a small-molecule inhibitor screen we identify signaling events that promote specifically IL-4-induced and not IFNγ/LPS-induced macrophage polarization and found that the MEK inhibitor trametinib and the HDAC inhibitor panobinostat potently prevent M2-type macrophage polarization without inhibiting M1-type polarization. In contrast, selective B-Raf inhibition promotes M2-type polarization. Trametinib and panobinostat also blocked M2-type macrophage polarization and concomitantly angiogenesis and fibrosis in models of wound healing and neovascular age-related macular degeneration in vivo. Thus, these pharmacologic inhibitors could be utilized therapeutically to selectively block IL4-induced macrophage polarization and reduce pathologic angiogenesis and fibrosis.
Project description:Macrophages polarize towards different subpopulations with distinct and partly antagonistic functions in various diseases. IFNγ/LPS-polarized M1-type macrophages can have antiangiogenic activity, whereas IL-4-induced M2-type macrophages can be proangiogenic and profibrotic. Therapeutic strategies to inhibit M2-type polarization while promoting M1-type polarization could serve to inhibit pathological angiogenesis and fibrosis. Here, by combining global quantitative time-course proteomics and phosphoproteomics with a small-molecule inhibitor screen we identify signaling events that promote specifically IL-4-induced and not IFNγ/LPS-induced macrophage polarization and found that the MEK inhibitor trametinib and the HDAC inhibitor panobinostat potently prevent M2-type macrophage polarization without inhibiting M1-type polarization. In contrast, selective B-Raf inhibition promotes M2-type polarization. Trametinib and panobinostat also blocked M2-type macrophage polarization and concomitantly angiogenesis and fibrosis in models of wound healing and neovascular age-related macular degeneration in vivo. Thus, these pharmacologic inhibitors could be utilized therapeutically to selectively block IL4-induced macrophage polarization and reduce pathologic angiogenesis and fibrosis.
Project description:Using the highly sensitive miRNA microarray, we screened 107 up-regulated microRNAs and 33 down-regulated microRNAs in the INT-HA-induced M2-like macrophages and we explored the functions of these miRNAs in macrophage polarization. The enrichment results indicated that these miRNAs might participate in the process of macrophage polarization. Furthermore, the quantitative real-time polymerase chain reaction results showed that miR-935 may play an important role in M2 macrophage polarization which was activated by INT-HA.
Project description:Clear cell renal cell carcinoma (ccRCC), a frequent urinary system tumor, is known for its poor patient outcomes. Here, we demonstrate that LINC00887 overexpression in ccRCC drives M2 macrophage polarization via aerobic glycolysis activation and lactate accumulation. Mechanistically, LINC00887 interacted with UPF1, leading to reduced UPF1 levels. UPF1, which is low in ccRCC, restrained cell vitality, proliferation, migration, invasion, and cell cycle progression in ccRCC cells, and stimulated apoptosis. Additionally, UPF1 was found to be negatively correlated with aerobic glycolysis in ccRCC, repressing glycolytic activity and M2 macrophage polarization while bolstering the tumor-killing potential of CD8+ T cells. By combining bioinformatics with experimental data from ccRCC single-cell RNA sequencing, we've demonstrated that reduced UPF1 expression can reprogram glycolytic metabolism, leading to an improved immunosuppressive tumor microenvironment.
Project description:Abstract Background: M2 macrophages (M2) play a crucial role in the development of colorectal cancer (CRC). The elevated expression of TIPE in CRC is closely associated with CRC progression; however, the impact of TIPE on M2 polarization remains unclear. Methods: IHC, WB, flow cytometry, qPCR, and IF were used to verify the promoting effect of TIPE on M2 polarization in specimens and in vitro co-culture models. RNA-seq, ELISA, and WB were used to identify the secreted proteins regulated by TIPE and the underlying mechanism. CCK-8 assay, wound healing assay, and transwell invasion assay were used to verify the effect of M2 on CRC. In vivo experiments were used to identify the promoting effect of TIPE on M2 polarization. Results: In this study, we first demonstrated that the abnormal overexpression of TIPE in CRC promotes M2 polarization and is correlated with a poor prognosis for CRC patients. We then found that TIPE activates the P38 MAPK signaling pathway in CRC to regulate TGFΒ2 secretion and, furthermore, induces M2 macrophage polarization. In subcutaneous and lung xenograft models of CRC in nude mice, we found that TIPE regulates TGFΒ2 secretion from human CRC to promote M2 polarization of murine macrophages. Conclusion: These results indicate that TIPE is a potent oncogene for inducing M2 polarization. It is suggested that all cancers with high TIPE expression need to be alert to the production and infiltration of M2 macrophages, which provides a new perspective for cancer treatment.