SAC Vice-Administrator Chen Hongjun met with the delegation of the Standards PoLicy Division, Korean Agency for TechnoLogy and Standards （KATS） on March 20, 2018.
目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化...目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化浊解毒方高剂量组(高剂量组)、益气活血化浊解毒方高剂量+2,5-己酮可可碱(DMXAA)组(高剂量+DMXAA组),每组15只。另取15只正常大鼠为Sham组。低剂量组大鼠灌胃0.2 mL 27.5 g/mL的益气活血化浊解毒方药液,高剂量组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液,高剂量+DMXAA组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液和25 mg/kg的DMXAA,Sham组和缺血性脑卒中组给予等体积生理盐水代替药物;除高剂量+DMXAA组外,其余组大鼠再给予等体积二甲基亚砜溶液灌胃,1 d/次,连续4周。Zea-Longa评分和网屏试验评分评估大鼠神经功能和神经行为学;检测大鼠脑组织含水量;TTC染色法检测大鼠脑梗死面积;酶联免疫吸附试验检测大鼠血清LDH和NGF水平和缺血侧皮层组织TNF-α、IL-6、IL-10水平;HE染色观察大鼠缺血侧皮层组织病理变化;Western blotting检测大鼠缺血侧皮层组织iNOS、Iba1、CD68、CD40、CD206、Arg-1、Cleaved caspase-3、KAT3B、STING蛋白表达。结果缺血性脑卒中组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于Sham组(P<0.05),低剂量组和高剂量组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均低于缺血性脑卒中组(P<0.05),高剂量+DMXAA组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于高剂量组(P<0.05)。缺血性脑卒中组大鼠NGF和IL-10水平低于Sham组,LDH、TNF-α和IL-6水平高于Sham组(P<0.05);低剂量组和高剂量组大鼠NGF和IL-10水平高于缺血性脑卒中组,LDH、TNF-α和IL-6水平低于缺血性脑卒中组(P<0.05);高剂量+DMXAA组大鼠NGF和IL-10水平低于高剂量组,LDH、TNF-α和IL-6高于高剂量组(P<0.05)。与缺血性脑卒中组比较,低剂量组和高剂量组缺血侧皮层神经元形态有明显改善。缺血性脑卒中组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比Sham组高,CD206和Arg-1蛋白相对表达量比Sham组低(P<0.05);低剂量组和高剂量组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比缺血性脑卒中组低,CD206和Arg-1蛋白相对表达量比缺血性脑卒中组高(P<0.05);高剂量+DMXAA组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比高剂量组高,CD206和Arg-1蛋白相对表达量比高剂量组低(P<0.05)。结论益气活血化浊解毒方可调节缺血性脑卒中大鼠小胶质细胞极化,减轻神经炎症和神经损伤,改善神经功能,可能与抑制KAT3B/STING轴有关。展开更多
BACKGROUND Immune checkpoint inhibitors(ICIs)are effective cancer treatments;however,a significant proportion of colorectal cancer(CRC)patients exhibit limited re-sponses to ICI therapy.KAT6A has been strongly associa...BACKGROUND Immune checkpoint inhibitors(ICIs)are effective cancer treatments;however,a significant proportion of colorectal cancer(CRC)patients exhibit limited re-sponses to ICI therapy.KAT6A has been strongly associated with cancer initiation and progression.AIM To examine the role of KAT6A in CRC progression and immune evasion.METHODS The functional role of KAT6A was evaluated through genetic knockdown,pharmacological inhibition(WM-3835),and CRISPR/dCas9-mediated epigenetic editing in CRC cells.T cell-mediated apoptosis was assessed using co-culture models,and H3K23pr was measured via chromatin immunoprecipitation assays.PD-L1 expression at mRNA and protein levels was analyzed under KAT6A knockdown conditions.RESULTS KAT6A suppression reduced CRC cell proliferation,invasion,and migration.Pharmacological or epigenetic disruption of KAT6A phenocopied these effects,with dose-dependent reductions in H3K23pr(28.4%residual at 10μM)and PD-L1 expression.KAT6A knockdown enhanced T cell-mediated apoptosis,evidenced by increased expression of granzyme B and perforin.Mechanistically,KAT6A loss decreased H3K23pr and reduced RNA polymerase II occupancy on the PD-L1 promoter,leading to suppressed PD-L1 transcription.CRISPR/dCas9-mediated H3K23pr editing at the PD-L1 promoter directly modulated immune evasion,confirming its causal role.Overexpression of PD-L1 mitigated the inhibitory effects of KAT6A knockdown on CRC progression and immune evasion.CONCLUSION KAT6A drives CRC progression and immune evasion by promoting histone H3 propionylation to epigenetically activate PD-L1 expression.Targeting KAT6A or its downstream H3K23pr-PD-L1 axis represents a promising therapeutic strategy to overcome ICI resistance in CRC.展开更多
文摘SAC Vice-Administrator Chen Hongjun met with the delegation of the Standards PoLicy Division, Korean Agency for TechnoLogy and Standards （KATS） on March 20, 2018.
文摘目的探讨益气活血化浊解毒方调控KAT3B/STING轴介导的小胶质细胞极化对缺血性脑卒中大鼠神经功能的影响。方法复制缺血性脑卒中大鼠模型。将复制成功的60只大鼠分为缺血性脑卒中组、益气活血化浊解毒方低剂量组(低剂量组)、益气活血化浊解毒方高剂量组(高剂量组)、益气活血化浊解毒方高剂量+2,5-己酮可可碱(DMXAA)组(高剂量+DMXAA组),每组15只。另取15只正常大鼠为Sham组。低剂量组大鼠灌胃0.2 mL 27.5 g/mL的益气活血化浊解毒方药液,高剂量组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液,高剂量+DMXAA组大鼠灌胃0.2 mL 55.0 g/mL的益气活血化浊解毒方药液和25 mg/kg的DMXAA,Sham组和缺血性脑卒中组给予等体积生理盐水代替药物;除高剂量+DMXAA组外,其余组大鼠再给予等体积二甲基亚砜溶液灌胃,1 d/次,连续4周。Zea-Longa评分和网屏试验评分评估大鼠神经功能和神经行为学;检测大鼠脑组织含水量;TTC染色法检测大鼠脑梗死面积;酶联免疫吸附试验检测大鼠血清LDH和NGF水平和缺血侧皮层组织TNF-α、IL-6、IL-10水平;HE染色观察大鼠缺血侧皮层组织病理变化;Western blotting检测大鼠缺血侧皮层组织iNOS、Iba1、CD68、CD40、CD206、Arg-1、Cleaved caspase-3、KAT3B、STING蛋白表达。结果缺血性脑卒中组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于Sham组(P<0.05),低剂量组和高剂量组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均低于缺血性脑卒中组(P<0.05),高剂量+DMXAA组大鼠Zea-Longa评分和网屏试验评分、脑组织含水量、脑梗死面积百分比均高于高剂量组(P<0.05)。缺血性脑卒中组大鼠NGF和IL-10水平低于Sham组,LDH、TNF-α和IL-6水平高于Sham组(P<0.05);低剂量组和高剂量组大鼠NGF和IL-10水平高于缺血性脑卒中组,LDH、TNF-α和IL-6水平低于缺血性脑卒中组(P<0.05);高剂量+DMXAA组大鼠NGF和IL-10水平低于高剂量组,LDH、TNF-α和IL-6高于高剂量组(P<0.05)。与缺血性脑卒中组比较,低剂量组和高剂量组缺血侧皮层神经元形态有明显改善。缺血性脑卒中组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比Sham组高,CD206和Arg-1蛋白相对表达量比Sham组低(P<0.05);低剂量组和高剂量组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比缺血性脑卒中组低,CD206和Arg-1蛋白相对表达量比缺血性脑卒中组高(P<0.05);高剂量+DMXAA组大鼠缺血侧皮层组织中iNOS、Iba1、CD68和CD40蛋白相对表达量比高剂量组高,CD206和Arg-1蛋白相对表达量比高剂量组低(P<0.05)。结论益气活血化浊解毒方可调节缺血性脑卒中大鼠小胶质细胞极化,减轻神经炎症和神经损伤,改善神经功能,可能与抑制KAT3B/STING轴有关。
文摘BACKGROUND Immune checkpoint inhibitors(ICIs)are effective cancer treatments;however,a significant proportion of colorectal cancer(CRC)patients exhibit limited re-sponses to ICI therapy.KAT6A has been strongly associated with cancer initiation and progression.AIM To examine the role of KAT6A in CRC progression and immune evasion.METHODS The functional role of KAT6A was evaluated through genetic knockdown,pharmacological inhibition(WM-3835),and CRISPR/dCas9-mediated epigenetic editing in CRC cells.T cell-mediated apoptosis was assessed using co-culture models,and H3K23pr was measured via chromatin immunoprecipitation assays.PD-L1 expression at mRNA and protein levels was analyzed under KAT6A knockdown conditions.RESULTS KAT6A suppression reduced CRC cell proliferation,invasion,and migration.Pharmacological or epigenetic disruption of KAT6A phenocopied these effects,with dose-dependent reductions in H3K23pr(28.4%residual at 10μM)and PD-L1 expression.KAT6A knockdown enhanced T cell-mediated apoptosis,evidenced by increased expression of granzyme B and perforin.Mechanistically,KAT6A loss decreased H3K23pr and reduced RNA polymerase II occupancy on the PD-L1 promoter,leading to suppressed PD-L1 transcription.CRISPR/dCas9-mediated H3K23pr editing at the PD-L1 promoter directly modulated immune evasion,confirming its causal role.Overexpression of PD-L1 mitigated the inhibitory effects of KAT6A knockdown on CRC progression and immune evasion.CONCLUSION KAT6A drives CRC progression and immune evasion by promoting histone H3 propionylation to epigenetically activate PD-L1 expression.Targeting KAT6A or its downstream H3K23pr-PD-L1 axis represents a promising therapeutic strategy to overcome ICI resistance in CRC.
