Apoptotic vesicles(ApoVs)are membrane structures formed during cell apoptosis and play crucial roles in homeostasis maintenance,signal transduction,and immune regulation.Importantly,ApoVs inherit the properties and co...Apoptotic vesicles(ApoVs)are membrane structures formed during cell apoptosis and play crucial roles in homeostasis maintenance,signal transduction,and immune regulation.Importantly,ApoVs inherit the properties and contents of parental cells that show great potential in the diagnosis and treatment of diseases.Monitoring the formation process of ApoVs(such as quantity,morphological changes,release rules,etc.)can reveal the regulatory mechanism of apoptosis,and is also helpful for optimizing the preparation and application of ApoVs.However,due to the limitations of existing technologies,the formation processes of ApoVs have been challenging to precisely and entirely capture.Herein,we subtly constructed a versatile AIEgen(ADTP)that could induce ApoVs production and in situ monitor the formation process,and it was successfully applied to explore the formation mechanism of ApoVs.ADTP specifically targeted the plasma membrane,and it could effectively induce apoptosis under laser irradiation,so it was able to dynamically monitor the entire formation process of ApoVs and had validated ApoVs formation from membrane protrusions(including filopodia,tunneling nanotubes,and retraction fibers).Further investigation revealed that ApoVs derived from membrane protrusions with different components exhibited significant heterogeneity.Additionally,the nearinfrared emission characteristic of ADTP was compatible with the stimulated emission depletion(STED)microscopy equipped with a 775nmdepletion laser,enabling high-resolution visualization of detailed dynamic changes inmembrane protrusions during ApoVs formation.This work provided powerful tools for tracking the entire ApoVs formation process and also offered crucial scientific evidence for revealing the ApoVs formation mechanism.展开更多
Background Effective therapies for Alzheimer’s disease(AD)remain to be developed.APOE4 is the strongest genetic risk factor for late-onset AD.Pericyte degeneration and blood–brain barrier(BBB)disruption are thought ...Background Effective therapies for Alzheimer’s disease(AD)remain to be developed.APOE4 is the strongest genetic risk factor for late-onset AD.Pericyte degeneration and blood–brain barrier(BBB)disruption are thought to be early biomarkers of AD and contribute to cognitive decline in APOE4 carriers,representing potential therapeutic targets.Our previous studies have shown that pericyte transplantation is one of the most effective strategies for BBB restoration,exhibiting great therapeutic potential for APOE4-related BBB damage and AD phenotypes.Methods APOE4/4 mice were treated with pericytes derived from APOE3/3 human induced pluripotent stem cells(hiPSCs).Behavioral tests,AD pathologies,and BBB integrity were assessed.Subsequently,temporal and spatial distribution of the transplanted pericytes was analyzed using tdTomato+lentivirus labeling.Next,therapeutic effects of apoptotic vesicles(ApoVs)generated from APOE3/3 pericytes were evaluated in APOE4/4 pericytes in vitro.Additionally,transcriptomic and proteomic profiling were performed to identify key effector molecules in pericyte-derived ApoVs.Finally,the therapeutic effects of ApoVs derived from pericytes were evaluated in APOE4/4 mice.Results Early,multiple transplantations of pericytes derived from APOE3/3 hiPSCs robustly rescued cognitive decline and AD pathologies,restored BBB integrity,and prevented in situ pericyte degeneration in aged APOE4/4 mice.Intriguingly,ApoVs released from the infused cells,rather than the transplanted pericytes,were predominantly distributed in the brain,which were ingested by in situ APOE4/4 pericytes and then promoted functional recovery.We further characterized insulin growth factor-2(IGF-2)as a key factor in APOE3/3 pericyte-derived ApoVs.Infusion of the in vitro generated ApoVs from APOE3/3 pericytes demonstrated distinct therapeutic effects in APOE4/4 mice,which were reversed by IGF2 knockout.Conclusions APOE3/3 pericytes or APOE3/3 pericyte-derived IGF2-rich ApoVs may offer promising therapeutic strategies for APOE4-associated AD.展开更多
基金supported by the National Natural Science Foundation of China(52350002,52150222,U2330106,and 52073163)the Special Fund of Taishan Scholars Project of Shandong Province(tsqnz20231253)+1 种基金Natural Science Foundation of Shandong Province(ZR2024QB200,ZR2023ZD41)Fundamental Research Funds for the Central Universities(2022JC003).
文摘Apoptotic vesicles(ApoVs)are membrane structures formed during cell apoptosis and play crucial roles in homeostasis maintenance,signal transduction,and immune regulation.Importantly,ApoVs inherit the properties and contents of parental cells that show great potential in the diagnosis and treatment of diseases.Monitoring the formation process of ApoVs(such as quantity,morphological changes,release rules,etc.)can reveal the regulatory mechanism of apoptosis,and is also helpful for optimizing the preparation and application of ApoVs.However,due to the limitations of existing technologies,the formation processes of ApoVs have been challenging to precisely and entirely capture.Herein,we subtly constructed a versatile AIEgen(ADTP)that could induce ApoVs production and in situ monitor the formation process,and it was successfully applied to explore the formation mechanism of ApoVs.ADTP specifically targeted the plasma membrane,and it could effectively induce apoptosis under laser irradiation,so it was able to dynamically monitor the entire formation process of ApoVs and had validated ApoVs formation from membrane protrusions(including filopodia,tunneling nanotubes,and retraction fibers).Further investigation revealed that ApoVs derived from membrane protrusions with different components exhibited significant heterogeneity.Additionally,the nearinfrared emission characteristic of ADTP was compatible with the stimulated emission depletion(STED)microscopy equipped with a 775nmdepletion laser,enabling high-resolution visualization of detailed dynamic changes inmembrane protrusions during ApoVs formation.This work provided powerful tools for tracking the entire ApoVs formation process and also offered crucial scientific evidence for revealing the ApoVs formation mechanism.
基金supported by grants from the National Key Research and Development Program of China(2021YFA1100603,2022YFA1104100,2024YFA1107900)the National Natural Science Foundation of China(82471462,82270566,32130046,82270886)+2 种基金the Pioneering Talents Project of Guangzhou Development Zone(2021-L029)the Key Scientific and Technological Program of Guangzhou City(2023B01J1002)the Sanming Project of Medicine in Shenzhen(SZSM202103012,SZSM202402019).
文摘Background Effective therapies for Alzheimer’s disease(AD)remain to be developed.APOE4 is the strongest genetic risk factor for late-onset AD.Pericyte degeneration and blood–brain barrier(BBB)disruption are thought to be early biomarkers of AD and contribute to cognitive decline in APOE4 carriers,representing potential therapeutic targets.Our previous studies have shown that pericyte transplantation is one of the most effective strategies for BBB restoration,exhibiting great therapeutic potential for APOE4-related BBB damage and AD phenotypes.Methods APOE4/4 mice were treated with pericytes derived from APOE3/3 human induced pluripotent stem cells(hiPSCs).Behavioral tests,AD pathologies,and BBB integrity were assessed.Subsequently,temporal and spatial distribution of the transplanted pericytes was analyzed using tdTomato+lentivirus labeling.Next,therapeutic effects of apoptotic vesicles(ApoVs)generated from APOE3/3 pericytes were evaluated in APOE4/4 pericytes in vitro.Additionally,transcriptomic and proteomic profiling were performed to identify key effector molecules in pericyte-derived ApoVs.Finally,the therapeutic effects of ApoVs derived from pericytes were evaluated in APOE4/4 mice.Results Early,multiple transplantations of pericytes derived from APOE3/3 hiPSCs robustly rescued cognitive decline and AD pathologies,restored BBB integrity,and prevented in situ pericyte degeneration in aged APOE4/4 mice.Intriguingly,ApoVs released from the infused cells,rather than the transplanted pericytes,were predominantly distributed in the brain,which were ingested by in situ APOE4/4 pericytes and then promoted functional recovery.We further characterized insulin growth factor-2(IGF-2)as a key factor in APOE3/3 pericyte-derived ApoVs.Infusion of the in vitro generated ApoVs from APOE3/3 pericytes demonstrated distinct therapeutic effects in APOE4/4 mice,which were reversed by IGF2 knockout.Conclusions APOE3/3 pericytes or APOE3/3 pericyte-derived IGF2-rich ApoVs may offer promising therapeutic strategies for APOE4-associated AD.
