Project description:Aplastic anaemia (AA) is a disease that shows complex pathogenesis involving multiple immune factors. While immunosuppressive therapies such as cyclosporine can effectively control AA, they may be ineffective in certain patients or those with relapse. Therefore, new treatments are needed. Among the targets for these treatments, the Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway regulates inflammatory cytokines and immune activation. Ruxolitinib, a JAK1/2 inhibitor, reduces T-cell activation and the associated inflammatory response and improves AA disease status in mice. However, its mechanism of action is unclear; thus, further research is needed before its clinical use. We previously showed increased pyroptosis in patients with severe AA (SAA) and that macrophage pyroptosis is an important factor in immune activation. The current study investigated the interaction of ruxolitinib with macrophages and whether the drug could treat SAA by improving pyroptosis levels. We induced differentiation of the THP-1 human monocyte cell line into macrophages in vitro and then induced pyroptosis. After constructing a macrophage pyroptosis model, treatment with different concentrations of ruxolitinib was administered. The results showed that ruxolitinib reduced the levels of pyroptosis and inflammatory-related factors. A mouse model of SAA was validated. In conclusion, ruxolitinib may treatment affects SAA by reducing the level of macrophage pyroptosis.
2025-03-18 | GSE291850 | GEO
Project description:Whole exome sequencing of aplastic anaemia
Project description:Background and methods: Ruxolitinib (RUX), a Jak1/2 inhibitor, has been reported to attenuate murine bone marrow failure recently. Its potential toxocicty of anemia and thrombocytopenia in human remains a concern. We tested the toxicity of ruxolitinib on hematopoiesis in normal mice by feeding the mice with RUX chow for 3 months. Bone marrow Lin-CD117+ cells were sorted from treated or untreated mice. RNA-Seq and analysis was performed using SMART-Seq mRNA LP Kit (Takara) and the Illumina Novaseq6000, according to the Institute's protocols. Results: Ruxolitinib reduced RBC and lymphocytes, but did not affect NEU and PLT in normal mice. RNA sequencing demonstrated that HSPCs from RUX-treated and untreated mice had overlapped transcriptome distribution in multiple dimentional scalling plot, indicating overall similarity at molecular level. Conclusion: Our results demonstrate that ruxolitinib has minimal toxicity on hematopoiesis in normal mice.
Project description:Bulk RNA-seq data of Lin-CD34+ hematopoietic stem and progenitor cells derived from bone marrow of healthy donors and untreated aplastic anemia patients
Project description:Pyroptosis is a recently discovered form of lytic cell death that is characterized by cell swelling and formation of pores and large bubbles on the plasma membrane. We find radiation could induce pyroptosis in human colorectal cancer HCT116 cells, and irradiation induces pyroptosis in mouse normal intestine MODEK cells only after 36 hours and over 8.0 Gy. In view of the significant pyroptosis effect of MODE-K cells, we selected 4Gy and 12Gy, 24h and 48h for experiments. After ONT full-length transcriptome sequencing, the cell pyroptosis model can screen out the differential genes related to cell pyroptosis, which is helpful to further explore the mechanism of pyroptosis of MODE-K cells.
Project description:Biomimetic nanococrystals are prepared using the preferred combination of mitoxantrone and gambogic acid for rational drug delivery. The unique mechanism of obtained nanococrystals regulating pyroptosis genes through ribosomal stress is revealed by TMT-proteomics of 4T1 cells.
Project description:Immune cells play critical roles in causing host damage in various diseases, including infectious diseases. However, there is a paucity of human models to study immune-mediated host damage. Here, we systematically analyzed the changes in immune cell composition for COVID-19 pancreatic autopsy samples using the GeoMx spatial RNA and protein platform and found accumulation of proinflammatory macrophages. Single cell RNA-seq analysis of human islets upon SARS-CoV-2 or Coxsackievirus B4 (CVB4) infection identified the activation of proinflammatory macrophages and pyroptosis in β cells. To distinguish whether the virus or proinflammatory macrophages mediated β cell pyroptosis, we developed human pluripotent stem cell (hPSC)-derived vascularized macrophage-islet (VMI) organoids, containing pancreatic endocrine cells, endothelial cells, and macrophages. Both pancreatic β cells and endothelial cells in VMI organoids demonstrated improved marker expression and function compared to separately cultured cells. Additionally, proinflammatory macrophages in VMI organoids were found to induce β cell pyroptosis. Further mechanistic studies identified TNFSF12-TNFRSF12A as a contributor to proinflammatory macrophage-mediated β cell pyroptosis. Together, this study established a hPSC-derived VMI organoid as a valuable tool for studying immune cell-mediated host damage and uncovered previous unknown mechanism of β cell damage in viral infection.
Project description:Immune cells play critical roles in causing host damage in various diseases, including infectious diseases. However, there is a paucity of human models to study immune-mediated host damage. Here, we systematically analyzed the changes in immune cell composition for COVID-19 pancreatic autopsy samples using the GeoMx spatial RNA and protein platform and found accumulation of proinflammatory macrophages. Single cell RNA-seq analysis of human islets upon SARS-CoV-2 or Coxsackievirus B4 (CVB4) infection identified the activation of proinflammatory macrophages and pyroptosis in β cells. To distinguish whether the virus or proinflammatory macrophages mediated β cell pyroptosis, we developed human pluripotent stem cell (hPSC)-derived vascularized macrophage-islet (VMI) organoids, containing pancreatic endocrine cells, endothelial cells, and macrophages. Both pancreatic β cells and endothelial cells in VMI organoids demonstrated improved marker expression and function compared to separately cultured cells. Additionally, proinflammatory macrophages in VMI organoids were found to induce β cell pyroptosis. Further mechanistic studies identified TNFSF12-TNFRSF12A as a contributor to proinflammatory macrophage-mediated β cell pyroptosis. Together, this study established a hPSC-derived VMI organoid as a valuable tool for studying immune cell-mediated host damage and uncovered previous unknown mechanism of β cell damage in viral infection.
Project description:Immune cells play critical roles in causing host damage in various diseases, including infectious diseases. However, there is a paucity of human models to study immune-mediated host damage. Here, we systematically analyzed the changes in immune cell composition for COVID-19 pancreatic autopsy samples using the GeoMx spatial RNA and protein platform and found accumulation of proinflammatory macrophages. Single cell RNA-seq analysis of human islets upon SARS-CoV-2 or Coxsackievirus B4 (CVB4) infection identified the activation of proinflammatory macrophages and pyroptosis in β cells. To distinguish whether the virus or proinflammatory macrophages mediated β cell pyroptosis, we developed human pluripotent stem cell (hPSC)-derived vascularized macrophage-islet (VMI) organoids, containing pancreatic endocrine cells, endothelial cells, and macrophages. Both pancreatic β cells and endothelial cells in VMI organoids demonstrated improved marker expression and function compared to separately cultured cells. Additionally, proinflammatory macrophages in VMI organoids were found to induce β cell pyroptosis. Further mechanistic studies identified TNFSF12-TNFRSF12A as a contributor to proinflammatory macrophage-mediated β cell pyroptosis. Together, this study established a hPSC-derived VMI organoid as a valuable tool for studying immune cell-mediated host damage and uncovered previous unknown mechanism of β cell damage in viral infection.