基于陽(yáng)離子聚合物的仿生藥物載體腫瘤免疫治療中的研究
發(fā)布時(shí)間：2018-07-13 12:46
【摘要】：如何將腫瘤新方法安全有效的應(yīng)用到腫瘤治療中,是腫瘤治療面臨的重大挑戰(zhàn)。納米技術(shù)的發(fā)展使高分子納米藥物載體在此領(lǐng)域擔(dān)任了至關(guān)重要的角色。基于環(huán)糊精-PEI的陽(yáng)離子聚合物作為載藥體系生物相容性好,且攜載基因轉(zhuǎn)染效率較高,但基于環(huán)糊精-PEI陽(yáng)離子聚合物在腫瘤多藥耐藥和腫瘤免疫治療方面應(yīng)用研究較少。因此,本論文對(duì)陽(yáng)離子聚合物及其仿生材料在腫瘤免疫治療、化療中的應(yīng)用作了以下研究,主要分以下三部分:第一部分,以γ-羥丙基-環(huán)糊精偶聯(lián)低分子量PEI600(HP)為主體,金剛烷甲酸修飾的阿霉素(AD)為客體經(jīng)主客自組裝為HPAD超分子納米材料,通過(guò)靜電作用攜載ETS1siRNA。形成內(nèi)核為疏水藥物,外殼攜載siRNA具有核殼結(jié)構(gòu)的協(xié)同給藥體系(HPAD/siRNA)。HPAD/siETS1經(jīng)細(xì)胞內(nèi)吞后進(jìn)入到溶酶體,通過(guò)質(zhì)子海綿效應(yīng)進(jìn)行溶酶體逃逸,由于溶酶體酸性環(huán)境使藥物和siRNA的釋放,發(fā)揮不同功能。ETS1 siRNA通過(guò)干擾ETS1蛋白表達(dá)降低MDR1表達(dá)水平,以加強(qiáng)乳腺癌耐藥細(xì)胞藥物敏感性,降低阿霉素的泵出。第二部分,設(shè)計(jì)將β-環(huán)糊精偶聯(lián)低分子量PEI600(CP)與DNA靜電結(jié)合后形成陽(yáng)離子聚合物納米材料(CP/DNA),用CP/DNA包被VEGFR2重組的減毒沙門氏菌(NP/SAL)構(gòu)建一種新型的腫瘤口服疫苗,并對(duì)NP/SAL疫苗進(jìn)行了活力檢測(cè)、形態(tài)學(xué)觀察、體外細(xì)胞轉(zhuǎn)染水平評(píng)估及體內(nèi)免疫原性、抗腫瘤能力等研究。實(shí)驗(yàn)證明NP/SAL通過(guò)口服后在NP層的保護(hù)下,減毒沙門氏菌免受胃酸環(huán)境消化,且聚陽(yáng)離子材料可以通過(guò)"質(zhì)子海綿效應(yīng)"導(dǎo)致溶酶體腫脹及膜的破裂,從而有助于細(xì)菌逃離吞噬體而到達(dá)腸粘膜部位。NP/SAL攜帶VEGFR2作為口服DNA疫苗能抑制腫瘤血管的形成,有利于抑制腫瘤的生長(zhǎng)與轉(zhuǎn)移。體內(nèi)實(shí)驗(yàn)結(jié)果證明NP/SAL疫苗能刺激免疫系統(tǒng)產(chǎn)生T細(xì)胞和細(xì)胞因子,進(jìn)而能影響VEGF通路,降低血管的形成,最終抑制腫瘤生長(zhǎng)。第三部分,將癌細(xì)胞膜(CM)和細(xì)菌S層蛋白(SLP)與陽(yáng)離子聚合物HPAD結(jié)合,制備了一種新型仿生納米材料S-CM-HPAD用于黑色素瘤的免疫治療和化療的聯(lián)合治療。該仿生材料以合成材料HPAD為核,以黑色素瘤細(xì)胞膜為外殼形成核殼結(jié)構(gòu),表面自組裝乳酸桿菌的S層蛋白(SLP)。在體內(nèi)外實(shí)驗(yàn)證明了S-CM-HPAD具有癌細(xì)胞膜的特征蛋白,能靶向同源腫瘤,增加藥物DOX療效。同時(shí),具有癌細(xì)胞膜上抗原,在免疫佐劑SLP的作用下能刺激機(jī)體產(chǎn)生抗腫瘤的CD4CD8T細(xì)胞和白介素-12、腫瘤壞死因子α、干擾素γ等細(xì)胞因子,發(fā)揮抗腫瘤免疫效應(yīng)。通過(guò)對(duì)小鼠體重和生存率的觀察,證明了 S-CM-HPAD生物相容性好,安全性高。
[Abstract]:How to safely and effectively apply the new tumor method to tumor treatment is a major challenge for tumor treatment. With the development of nanotechnology, polymer nano-drug carriers play an important role in this field. Cationic polymer based on cyclodextrin-PEI has good biocompatibility and high transfection efficiency as a drug carrier system. However, the application of cyclodextrin-PEI cationic polymer in tumor multidrug resistance and tumor immunotherapy is less. Therefore, the application of cationic polymer and its biomimetic materials in tumor immunotherapy and chemotherapy was studied as follows: the first part was composed of 緯 -hydroxypropyl-cyclodextrin coupled low molecular weight PEI600 (HP). Adriamycin (AD) modified by adriamycin (AD) was self-assembled by host and guest to form Hpad supramolecular nanomaterials. ETS1siRNA was carried by electrostatic interaction. The core was formed as a hydrophobic drug, and the shell carried core-shell siRNA with core-shell structure (HPAD / siRNA). HPAD / siETS1 entered the lysosome after endocytosis and escaped from lysosome by proton sponge effect. The release of drugs and siRNA was due to the acidic environment of lysosome. By interfering with the expression of ETS1 protein, the expression of MDR1 was reduced by different functions of ETS1 siRNA, in order to enhance the drug sensitivity of breast cancer cells and reduce the pump of doxorubicin. In the second part, 尾 -cyclodextrin coupled low molecular weight PEI600 (CP) was designed to form cationic polymer nanomaterials (CP-DNA) after electrostatic binding with DNA. A novel oral tumor vaccine was constructed by encapsulating the recombinant attenuated Salmonella mutans VEGFR2 (NPP / SAL) with CPR / DNA. The NPP / Sal vaccine was tested for its viability, morphological observation, in vitro cell transfection level evaluation, in vivo immunogenicity and anti-tumor ability. The results show that NPP / Sal can protect attenuated Salmonella from gastric acid environment after oral administration, and that polycationic materials can cause lysosome swelling and membrane rupture through "proton sponge effect". It is helpful for bacteria to escape phagocytosis and reach intestinal mucosa. NPP / Sal carrying VEGFR2 as oral DNA vaccine can inhibit tumor angiogenesis and inhibit tumor growth and metastasis. The results show that NPP Sal vaccine can stimulate the immune system to produce T cells and cytokines, which can affect the VEGF pathway, reduce the formation of blood vessels, and ultimately inhibit tumor growth. In the third part, a novel biomimetic nano-material S-CM-Hpad was prepared by combining cancer cell membrane (CM) and bacterial S layer protein (SLP) with cationic polymer Hpad for immunotherapy and chemotherapy of melanoma. The biomimetic material consisted of synthetic material Hpad and melanoma cell membrane to form core-shell structure and self-assemble S-layer protein (SLP) of Lactobacillus. In vitro and in vivo experiments proved that S-CM-Hpad has the characteristic protein of cancer cell membrane, which can target homologous tumor and increase the therapeutic effect of DOX. At the same time, with the antigen on the membrane of cancer cells, SLP can stimulate the production of anti-tumor CD4CD8T cells, interleukin-12, tumor necrosis factor 偽, interferon 緯 and other cytokines, play an anti-tumor immune effect. It was proved that S-CM-Hpad had good biocompatibility and high safety by observing the body weight and survival rate of mice.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：O631;TQ460.1
本文編號(hào)：2119414
[Abstract]:How to safely and effectively apply the new tumor method to tumor treatment is a major challenge for tumor treatment. With the development of nanotechnology, polymer nano-drug carriers play an important role in this field. Cationic polymer based on cyclodextrin-PEI has good biocompatibility and high transfection efficiency as a drug carrier system. However, the application of cyclodextrin-PEI cationic polymer in tumor multidrug resistance and tumor immunotherapy is less. Therefore, the application of cationic polymer and its biomimetic materials in tumor immunotherapy and chemotherapy was studied as follows: the first part was composed of 緯 -hydroxypropyl-cyclodextrin coupled low molecular weight PEI600 (HP). Adriamycin (AD) modified by adriamycin (AD) was self-assembled by host and guest to form Hpad supramolecular nanomaterials. ETS1siRNA was carried by electrostatic interaction. The core was formed as a hydrophobic drug, and the shell carried core-shell siRNA with core-shell structure (HPAD / siRNA). HPAD / siETS1 entered the lysosome after endocytosis and escaped from lysosome by proton sponge effect. The release of drugs and siRNA was due to the acidic environment of lysosome. By interfering with the expression of ETS1 protein, the expression of MDR1 was reduced by different functions of ETS1 siRNA, in order to enhance the drug sensitivity of breast cancer cells and reduce the pump of doxorubicin. In the second part, 尾 -cyclodextrin coupled low molecular weight PEI600 (CP) was designed to form cationic polymer nanomaterials (CP-DNA) after electrostatic binding with DNA. A novel oral tumor vaccine was constructed by encapsulating the recombinant attenuated Salmonella mutans VEGFR2 (NPP / SAL) with CPR / DNA. The NPP / Sal vaccine was tested for its viability, morphological observation, in vitro cell transfection level evaluation, in vivo immunogenicity and anti-tumor ability. The results show that NPP / Sal can protect attenuated Salmonella from gastric acid environment after oral administration, and that polycationic materials can cause lysosome swelling and membrane rupture through "proton sponge effect". It is helpful for bacteria to escape phagocytosis and reach intestinal mucosa. NPP / Sal carrying VEGFR2 as oral DNA vaccine can inhibit tumor angiogenesis and inhibit tumor growth and metastasis. The results show that NPP Sal vaccine can stimulate the immune system to produce T cells and cytokines, which can affect the VEGF pathway, reduce the formation of blood vessels, and ultimately inhibit tumor growth. In the third part, a novel biomimetic nano-material S-CM-Hpad was prepared by combining cancer cell membrane (CM) and bacterial S layer protein (SLP) with cationic polymer Hpad for immunotherapy and chemotherapy of melanoma. The biomimetic material consisted of synthetic material Hpad and melanoma cell membrane to form core-shell structure and self-assemble S-layer protein (SLP) of Lactobacillus. In vitro and in vivo experiments proved that S-CM-Hpad has the characteristic protein of cancer cell membrane, which can target homologous tumor and increase the therapeutic effect of DOX. At the same time, with the antigen on the membrane of cancer cells, SLP can stimulate the production of anti-tumor CD4CD8T cells, interleukin-12, tumor necrosis factor 偽, interferon 緯 and other cytokines, play an anti-tumor immune effect. It was proved that S-CM-Hpad had good biocompatibility and high safety by observing the body weight and survival rate of mice.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：O631;TQ460.1
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 馮珂珂,趙洪洋,邱惠,陳劍;表達(dá)VEGFR-2的減毒鼠傷寒沙門氏菌疫苗誘導(dǎo)特異性抗膠質(zhì)瘤血管免疫應(yīng)答[J];癌癥;2005年05期
2 ;Development of an oral DNA vaccine against MG7-Ag of gastric cancer using attenuated salmonella typhimurium as carrier[J];World Journal of Gastroenterology;2003年06期
，本文編號(hào)：2119414
本文鏈接：http://sikaile.net/shoufeilunwen/gckjbs/2119414.html
最近更新
教材專著
