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《现代肿瘤医学》[ISSN:1672-4992/CN:61-1415/R]

期数:

2022年14期

页码:

2515-2520

栏目:

论著（基础研究）

出版日期:

2022-06-12

文章信息/Info

Title:

Liposome assisted delivery of CTLA-4 siRNA inhibits the growth of renal cell carcinoma by activating systemic anti-tumor immune response

作者:

拜合提亚·阿扎提; 李前进; 刘 强; 王玉杰

新疆医科大学第一附属医院泌尿外科，新疆 乌鲁木齐 830054

Author(s):

Baihetiya·Azhati; LI Qianjin; LIU Qiang; WANG Yujie

Department of Urology,the First Affiliated Hospital of Xinjiang Medical University,Xinjiang Urumqi 830054，China.

关键词:

肾细胞癌; 脂质体; AS1411; siRNA; CTLA-4

Keywords:

renal cell carcinoma; liposome; AS1411; siRNA; CTLA-4

分类号:

R737.11

DOI:

10.3969/j.issn.1672-4992.2022.14.006

文献标识码:

A

摘要:

目的：探索携带CTLA-4 siRNA的适配子偶联脂质体颗粒是否可以激活肿瘤部位的抗肿瘤免疫反应，抑制肾细胞癌的生长。方法：采用薄膜水化法制备脂质体；使用透射电子显微镜观察脂质体的形态和结构；用Zetasizer测量Zeta电位；共孵育实验观察靶细胞对Lipo-siRNA的摄取；qPCR检测Lipo-siRNA对CTLA-4基因的沉默；小鼠移植瘤模型检测Lipo-siRNA的体内抑瘤能力；流式细胞术检测肿瘤浸润T细胞的激活状态；免疫荧光法检测肿瘤浸润T细胞的数目。结果：成功制备Lipo-siRNA，电镜结果显示其具有双层球状结构；Zetasizer测得其Zeta电位为(+20.53±2.66)mV；荧光显微镜观察结果表明Lipo-siRNA可以被靶细胞有效摄取，qPCR检测Lipo-siRNA可以显著降低CTLA-4基因的表达（P＜0.001）；小鼠移植瘤模型显示Lipo-siRNA较对照组而言可以显著抑制肿瘤生长（P＜0.001），降低肿瘤细胞中CTLA-4的表达（P＜0.001），提升肿瘤浸润T细胞的数量（P＜0.000 1），并且提高了肿瘤浸润T细胞中IL-2（P＜0.000 1）和IFN-γ（P＜0.000 1）的表达水平。结论：适配子偶联脂质体可以携带CTLA-4 siRNA靶向肿瘤细胞，激活肿瘤部位的抗肿瘤免疫反应，抑制肿瘤的生长，对肾细胞癌的治疗具有潜在的临床应用价值。

Abstract:

Objective:To explore whether aptamer-coupled liposome particles carrying CTLA-4 siRNA can activate anti-tumor immune response and inhibit the growth of renal cell carcinoma.Methods:Liposomes were prepared by thin film hydration method.The morphology and structure of liposomes were observed by transmission electron microscope.Zeta potential was measured by Zetasizer.The uptake of Lipo-siRNA by target cells was observed in co-incubation experiment.The silencing of Lipo-siRNA to CTLA-4 gene was detected by qPCR.The anti-tumor ability of Lipo-siRNA in vivo was detected by mouse transplanted tumor model.The activation of tumor infiltrating T cells was detected by flow cytometry,and the number of tumor infiltrating T cells was detected by immunofluorescence.Results:The Lipo-siRNA was successfully prepared.It had a double-layer spherical structure,and its Zeta potential was (+20.53±2.66)mV measured by Zetasizer.The results of fluorescence microscope showed that Lipo-siRNA could be effectively absorbed by target cells,and qPCR detection revealed that Lipo-siRNA could significantly reduce the expression of CTLA-4 gene（P＜0.001）.Compared with the control group,mouse transplanted tumor model showed that Lipo-siRNA could significantly inhibit tumor growth（P＜0.001）,reduce the expression of CTLA-4 in tumor cells（P＜0.001）,increase the number of tumor infiltrating T cells（P＜0.000 1）,and increase the expression levels of IL-2（P＜0.000 1） and IFN-γ（P＜0.000 1） in tumor infiltrating T cells.Conclusion:Aptamer-coupled liposomes can carry CTLA-4 siRNA to target tumor cells,activate anti-tumor immune response at tumor site and inhibit tumor growth,which has potential clinical value in the treatment of renal cell carcinoma.

参考文献/References

［1］ TAKEDA K,MURRAY G,VOHRA N，et al.A case of the world's largest renal cell carcinoma ［J］.IJU Case Rep,2021,4(1):49-52.
［2］ WANG Y,YANG J,ZHANG Q，et al.Extent and characteristics of immune infiltration in clear cell renal cell carcinoma and the prognostic value ［J］.Transl Androl Urol,2019,8(6):609-618.
［3］ ANTONELLI A,PALUMBO C,SANDRI M，et al.Renal function impairment below safety limits correlates with cancer-specific mortality in localized renal cell carcinoma:Results from a single-center study ［J］.Clin Genitourin Cancer,2020,18(4):e360-e367.
［4］ NAKAMURA H,TANAKA S,MIYANISHI K，et al.A case of hypervascular tumors in the liver and pancreas:Synchronous hepatocellular carcinoma and pancreatic metastasis from renal cell carcinoma 36 years after nephrectomy ［J］.Clin Case Rep,2021,9(2):932-937.
［5］ VARKARIS A,XU W,DAVIS RB，et al.Combining immune checkpoint and vegfr inhibition in favorable risk and elderly patients with metastatic renal cell carcinoma ［J］.Clin Genitourin Cancer,2020,18(3):179-184.
［6］ MIRY A,EL AISSAOUY W,RAHOU FZ，et al.Chromophobe renal cell carcinoma revealed by stercoral peritonitis:Case report ［J］.Tunis Med,2020,98(6):522-526.
［7］ PISTILLO MP,CAROSIO R,GRILLO F，et al.Phenotypic characterization of tumor ctla-4 expression in melanoma tissues and its possible role in clinical response to ipilimumab ［J］.Clin Immunol,2020,215:108428.
［8］ HE M,CHAI Y,QI J，et al.Remarkably similar ctla-4 binding properties of therapeutic ipilimumab and tremelimumab antibodies ［J］.Oncotarget,2017,8(40):67129-67139.
［9］ ROSSKOPF S,LEITNER J,ZLABINGER GJ，et al.Ctla-4 antibody ipilimumab negatively affects cd4(+) t-cell responses in vitro ［J］.Cancer Immunol Immunother,2019,68(8):1359-1368.
［10］ SAKAMURI D,GLITZA IC,BETANCOURT CUELLAR SL，et al.Phase i dose-escalation study of anti-ctla-4 antibody ipilimumab and lenalidomide in patients with advanced cancers ［J］.Mol Cancer Ther,2018,17(3):671-676.
［11］ KATO M,HUANG YY,MATSUO M，et al.Rnai-mediated knockdown of mouse melanocortin-4 receptor in vitro and in vivo,using an sirna expression construct based on the mir-187 precursor ［J］.Exp Anim,2017,66(1):41-50.
［12］ ALAGIA A,ERITJA R.Sirna and rnai optimization ［J］.Wiley Interdiscip Rev RNA,2016,7(3):316-329.
［13］ HE C,YUE H,XU L，et al.Sirna release kinetics from polymeric nanoparticles correlate with rnai efficiency and inflammation therapy via oral delivery ［J］.Acta Biomater,2020,103:213-222.
［14］ TENG Y,GIRVAN AC,CASSON LK，et al.As1411 alters the localization of a complex containing protein arginine methyltransferase 5 and nucleolin ［J］.Cancer Res,2007,67(21):10491-10500.
［15］ MASTRACCI L,FONTANA V,QUEIROLO P，et al.Response to ipilimumab therapy in metastatic melanoma patients:Potential relevance of ctla-4(+) tumor infiltrating lymphocytes and their in situ localization ［J］.Cancer Immunol Immunother,2020,69(4):653-662.
［16］ 李丽，陈公琰.CTLA-4在肺癌中的研究进展［J］.现代肿瘤医学，2021,29（02）：348-352. LI Li，CHEN Gongyan.Research progress of CTLA-4 in lung cancer ［J］.Modern Oncology,2021,29（02）：348-352.
［17］ LENS M,TESTORI A,FERUCCI PF.Ipilimumab targeting cd28-ctla-4 axis:New hope in the treatment of melanoma ［J］.Curr Top Med Chem,2012,12(1):61-66.
［18］ HUANG G,GAO Q,ZHAO Y，et al.A novel sirna validation system for functional screening of effective rnai targets in mammalian cells and development of a derivative lentivirus delivery system ［J］.Gene,2015,558(2):278-286.
［19］ KIM BS,CHUANOI S,SUMA T，et al.Self-assembly of sirna/peg-b-catiomer at integer molar ratio into 100 nm-sized vesicular polyion complexes (sirnasomes) for rnai and codelivery of cargo macromolecules ［J］.J Am Chem Soc,2019,141(8):3699-3709.
［20］ FAN Y,WANG Q,LIN G，et al.Combination of using prodrug-modified cationic liposome nanocomplexes and a potentiating strategy via targeted co-delivery of gemcitabine and docetaxel for cd44-overexpressed triple negative breast cancer therapy ［J］.Acta Biomater,2017,62:257-272.
［21］ KIM YB,ZHAO KT,THOMPSON DB，et al.An anionic human protein mediates cationic liposome delivery of genome editing proteins into mammalian cells ［J］.Nat Commun,2019,10(1):2905.
［22］ ZHANG H,YU N,CHEN Y，et al.Cationic liposome codelivering pi3k pathway regulator improves the response of brca1-deficient breast cancer cells to parp1 inhibition ［J］.J Cell Biochem,2019,120(8):13037-13045.
［23］ BANESHI M,DADFARNIA S,SHABANI AMH，et al.A novel theranostic system of as1411 aptamer-functionalized albumin nanoparticles loaded on iron oxide and gold nanoparticles for doxorubicin delivery ［J］.Int J Pharm,2019,564:145-152.

备注/Memo

备注/Memo:

吴阶平医学基金会临床科研专项资助基金（编号：320.6750.19094-43）

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更新日期/Last Update: 1900-01-01