目的:探讨溶质载体家族38成员2(solute carrier family 38 member 2,SLC38A2)对自然杀伤(natural killer,NK)细胞肿瘤杀伤活性的影响。方法:采用实时荧光定量PCR法检测谷氨酰胺缺失NK细胞和节食小鼠脾脏中谷氨酰胺相关氨基酸转运蛋白(SL...目的:探讨溶质载体家族38成员2(solute carrier family 38 member 2,SLC38A2)对自然杀伤(natural killer,NK)细胞肿瘤杀伤活性的影响。方法:采用实时荧光定量PCR法检测谷氨酰胺缺失NK细胞和节食小鼠脾脏中谷氨酰胺相关氨基酸转运蛋白(SLC38A2、SLC1A5、SLC7A5、SLC7A1和SLC1A4)mRNA的表达情况。通过慢病毒感染的方法将携带有SLC38A2基因重组慢病毒载体转入NK-92MI细胞使SLC38A2过表达。在SLC38A2过表达或谷氨酰胺浓度提高后,利用乳酸脱氢酶释放实验检测NK-92MI细胞对胰腺癌细胞PANC-1和肝癌细胞HUH-7的杀伤活性,并通过FCM法(FITC-Annexin V/7-AAD染色)检测NK-92MI细胞对PANC-1和HUH-7细胞凋亡的影响。最后,通过FCM法进一步检测NK-92MI细胞中杀伤效应分子干扰素-γ(interferon-γ,IFN-γ)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的表达水平以及脱颗粒标志分子CD107a的表达水平。结果:在缺失谷氨酰胺的培养条件下以及节食小鼠模型中获得的NK细胞中氨基酸转运蛋白SLC38A2 mRNA的表达显著增高(P<0.001和P<0.01)。成功构建SLC38A2过表达的NK-92MI细胞;SLC38A2过表达或谷氨酰胺处理均可以明显提升NK-92MI细胞对胰腺癌PANC-1和肝癌HUH-7细胞的杀伤能力(P均<0.05),同时对肿瘤细胞的凋亡具有明显的促进作用(P均<0.01)。流式细胞分析结果显示,过表达SLC38A2的NK-92MI细胞在杀伤肿瘤细胞的过程中,细胞内IFN-γ和TNF-α的含量均显著增加(P均<0.001),细胞膜表面CD107a的表达水平显著提高(P<0.001)。结论:氨基酸转运蛋白SLC38A2能显著增强NK细胞对肿瘤细胞的杀伤活性。展开更多
Objective:This study was aimed at investigating metabolic dysregulation in tumor-associated macrophages(TAMs)in breast cancer and developing a metabolically enhanced chimeric antigen receptor macrophage(CAR-M)strategy...Objective:This study was aimed at investigating metabolic dysregulation in tumor-associated macrophages(TAMs)in breast cancer and developing a metabolically enhanced chimeric antigen receptor macrophage(CAR-M)strategy to boost antitumor potency in solid tumors.Methods:Integrated scRNA-seq and metabolomic analyses were performed to characterize metabolic alterations in macrophages within the breast cancer tumor microenvironment(TME).According to the identified metabolic vulnerabilities,SLC38A2-overexpressing anti-HER2 CAR-Ms were engineered.Glutamine uptake and phagocytic activity were assessed to evaluate functional enhancement.Results:TAMs in breast cancer exhibited substantial metabolic dysregulation,particularly impaired glutamine metabolism accompanied by decreased expression of the glutamine transporter SLC38A2.Overexpression of SLC38A2 in anti-HER2 CAR-Ms,compared with conventional anti-HER2 CAR-Ms,enhanced glutamine uptake and markedly augmented phagocytosis of HER2+breast cancer cells.Conclusions:Metabolic engineering via SLC38A2 restored glutamine fitness and enhanced the antitumor activity of HER2-targeted CAR-Ms,thus providing a promising strategy to boost CAR-M–mediated tumor suppression in solid tumors.展开更多
基金supported by grants from the National Key R&D Program of China(Grant Nos.2021YFA1103000 and 2021YFA1302000)National Natural Science Foundation of China(Grant Nos.82572106,82125017,and 92359302)+7 种基金Guangdong Province Youth Top Talent Special Pillar Program 2024,Natural Science Foundation of Guangdong Province(Grant No.2314050001076)Guangdong Basic and Applied Basic Research Foundation(Grant Nos.2023A1515011033,2024B1515040006,and 2025A1515012431)Guangdong Major Project of Basic Research(Grant No.2023B0303000018)Guangzhou Science and Technology Plan Project(Grant No.2025B03J0063)Noncommunicable Chronic Diseases-National Science and Technology Major Project(Grant No.2025ZD0544000)Guangdong Provincial Clinical Research Center for Breast Diseases(Grant No.2023B110005)the Science and Technology Program of Guangzhou(Grant No.2024A04J6568)New Cornerstone Science Foundation through the New Cornerstone Investigator Program,and XPLORER PRIZE.
文摘Objective:This study was aimed at investigating metabolic dysregulation in tumor-associated macrophages(TAMs)in breast cancer and developing a metabolically enhanced chimeric antigen receptor macrophage(CAR-M)strategy to boost antitumor potency in solid tumors.Methods:Integrated scRNA-seq and metabolomic analyses were performed to characterize metabolic alterations in macrophages within the breast cancer tumor microenvironment(TME).According to the identified metabolic vulnerabilities,SLC38A2-overexpressing anti-HER2 CAR-Ms were engineered.Glutamine uptake and phagocytic activity were assessed to evaluate functional enhancement.Results:TAMs in breast cancer exhibited substantial metabolic dysregulation,particularly impaired glutamine metabolism accompanied by decreased expression of the glutamine transporter SLC38A2.Overexpression of SLC38A2 in anti-HER2 CAR-Ms,compared with conventional anti-HER2 CAR-Ms,enhanced glutamine uptake and markedly augmented phagocytosis of HER2+breast cancer cells.Conclusions:Metabolic engineering via SLC38A2 restored glutamine fitness and enhanced the antitumor activity of HER2-targeted CAR-Ms,thus providing a promising strategy to boost CAR-M–mediated tumor suppression in solid tumors.
