Synthetic Biology-Powered Cell Membrane-Derived Nanoparticles for Precision Theranostics

doi:10.12211/2096-8280.2025-069

Abstract

Abstract:

Cell membrane-derived nanoparticles (CNPs) integrate the biological characteristics of natural cell membranes (e.g., immune evasion, lesion targeting and immune modulation) with the tailorable physicochemical properties of synthetic nanomaterials, demonstrating significant advantages such as prolonged circulation, high biocompatibility, and specific targeting in disease diagnosis and treatment. However, their clinical application is limited by the inherent heterogeneity and functional limitations of natural membranes, including restricted targeting specificity, uncontrollable responsiveness, and lack of functionality. Synthetic biology provides innovative strategies to overcome these bottlenecks, driving a paradigm shift in CNPs from natural biomimicry to precise design. Genetic engineering enables precise editing of cell membrane protein expression via physical (e.g., electroporation, microinjection and gene gun), chemical (cationic lipids/polymers), and biological (viral vectors) strategies. Concurrently, metabolic engineering regulates the directional anchoring of functional moieties on cell membrane through manipulating cells' natural biosynthetic pathways, such as glycan (sialic acid and N-acetylgalactosamine (GalNAc) salvage pathways) and lipid (cytidine 5'-diphosphocholine pathway) metabolism. These approaches endow CNPs with enhanced targeting specificity, intelligent responsiveness (e.g., pH/enzyme/light-triggered drug release), and multifunctional synergy, enabling them to demonstrate significant therapeutic potential across diverse diseases including malignant tumors, cardiovascular diseases, and infectious diseases. In oncology, synthetic CNPs (SCNPs) deliver chemotherapeutics, radiotherapy sensitizers and contrast agents with tumor-homing specificity and enable innovative immunotherapies by presenting checkpoint inhibitors, tumor antigens or adjuvants. For cardiovascular diseases, SCNPs demonstrate remarkable inflammatory targeting and alleviation capabilities. In infectious diseases, SCNPs neutralize toxins, bacteria and viruses as broad-spectrum "nanosponges", while antigen-presenting SCNPs act as potent vaccines. Applications of SCNPs extend to autoimmune conditions, neurodegenerative disorders and bone-related diseases. Although challenges remain in safety assessment, scalable manufacturing, and regulatory frameworks, advances in artificial intelligence-assisted rational design, novel gene editing tools (e.g., prime editing) for safer genomic modifications, metabolic intervention technologies optimizations, alongside the establishment of standardized production platforms, are poised to bridge the gap between laboratory and clinic. Ultimately, synthetic biology-powered CNPs are anticipated to evolve into intelligent nano-theranostic platformsfacilitating precision medicine.

Key words: cell membrane-derived nanoparticles, synthetic biology, genetic engineering, metabolic engineering, precision medicine

摘要：

细胞膜纳米颗粒（cell membrane-derived nanoparticles， CNPs）能够有效整合天然细胞膜的生物学特性和纳米材料的理化性质，在疾病诊疗研究中展现出循环时间长、生物相容性好和靶向特异性强等优势；但其临床应用受限于天然膜的异质性和功能局限性。合成生物学为突破这一瓶颈提供了创新性策略，驱动CNPs实现从天然仿生到精准设计的范式转变。基因工程技术通过物理、化学及生物学手段精准编辑细胞膜蛋白表达，而代谢工程技术通过糖类和脂质代谢等通路实现细胞膜表面功能分子的定向锚定，从而赋予CNPs增强的靶向特异性、智能响应性与多功能协同性，使其在恶性肿瘤、心血管疾病和感染性疾病等多种疾病领域展现出巨大潜力。尽管在安全性评估、规模化生产和监管框架构建等方面仍面临挑战，但随着人工智能辅助设计、新型基因编辑和代谢介入技术的发展以及标准化生产平台的建立，合成生物学赋能的CNPs有望实现从实验室研究向临床应用的跨越，发展成为助力精准医疗的智能纳米诊疗平台。

关键词: 细胞膜纳米颗粒, 合成生物学, 基因工程, 代谢工程, 精准医疗

CLC Number:

Q816

HUANG Yang, LI Yiye, NIE Guangjun. Synthetic Biology-Powered Cell Membrane-Derived Nanoparticles for Precision Theranostics[J]. Synthetic Biology Journal, DOI: 10.12211/2096-8280.2025-069.

黄扬, 李一叶, 聂广军. 合成生物学赋能细胞膜纳米颗粒的精准诊疗[J]. 合成生物学, DOI: 10.12211/2096-8280.2025-069.

Figures/Tables 12

Fig. 1 Preparation of NCNPs. Created in BioRender. Huang Yang (2025)[The preparation of NCNPs usually involves three steps: (1) natural cell membrane extraction, mainly involves techniques such as hypotonic lysis, freeze-thawing, ultrasonic waves, and homogenization; (2) preparation of nanocores; (3) fusion of cell membrane and nanocore, mainly involves techniques such as extrusion, sonication, and electroporation.]

Fig. 2 Engineering strategies for SCNPs. Created in BioRender. Huang Yang (2025)[Mainly includes: (1) lipid insertion, which incorporates functional ligands onto natural cell membranes through a lipid anchor;(2) membrane hybridization, which derives the membrane from individual cell types and then fuse them or fuses different live cells and then derives membrane from the cell hybrids;(3) genetic engineering, which alters protein expression on the cell surfaces through selective gene editing and (4) metabolic engineering, which conjugates functional moieties with metabolic substrates to anchor them to the cell surfaces through natural metabolic pathways.]

Fig. 3 Strategies of gene transfection. Created in BioRender. Huang Yang (2025)[It mainly includes physical strategies (e.g., electricity/ultrasound/laser -assisted perforation, gene gun and microinjection), chemical strategies (e.g., cationic lipids/polymers -mediated transmembrane transport), and biological strategies (e.g., viral transduction).]

Table 1 Biomedical applications of nanoparticles with genetically engineered membranes

基因递送载体		细胞膜来源	特异表达蛋白	生物医学应用	参考文献
病毒载体	逆转录病毒载体	脂肪来源干细胞	CXCR4	通过CXCR4/SDF-1靶向炎症部位	[76]
	慢病毒载体	RAW 264.7	CCR2	通过CCR2/CCL2靶向炎症部位，阻断炎症信号，缓解炎症	[77]
		LX2	TRAIL	通过TRAIL-受体相互作用诱导肿瘤细胞凋亡	[80]
		神经干细胞	CXCR4	通过CXCR4/SDF-1靶向炎症部位，跨越血脑屏障	[82]
		CT26	HER2抗体	通过HER2抗体/HER靶向肿瘤部位，激活肿瘤免疫	[19]
		RAW 264.7	HA，RAGE	通过RAGE/S100A9靶向炎症受损心肌，缓解炎症和心肌损伤；通过HA实现内体逃逸	[81]
	腺病毒载体	DCs	MHC-I，PD 1，B7	通过MHC-I/PD-1/B7活化T细胞，激活肿瘤免疫	[79]
	腺病毒载体	神经干细胞	Lamp 2b-RVG	通过RVG/乙酰胆碱受体靶向神经细胞，跨越血脑屏障	[78]
非病毒载体	阳离子脂质体	B16-F10	卵清白蛋白，CD80	通过卵清白蛋白/CD80活化T细胞，激活肿瘤免疫	[31]
		B16-F10	HA	通过HA实现内体逃逸	[86]
		C1498	VLA-4	通过VLA-4/VCAM-1靶向炎症部位	[39]
		HEK293T	PD-1	通过PD-1/PD-L1靶向肿瘤部位，阻断免疫检查点，激活肿瘤免疫	[75]
	阳离子聚合物	4T1-Fluc	KillerRed	通过KillerRed实现光动力治疗，杀伤肿瘤	[73]
	阳离子聚合物	B16-F10	CD 47 KO/CRT	激活肿瘤免疫	[74]

Table 2 Biomedical applications and synthesis of nanoparticles with metabolically engineered membranes

代谢工程	生物合成途径	代谢底物	生物医学应用	参考文献
糖工程	唾液酸途径	N-乙酰甘露糖胺 ManNAc	插入N₃偶联肝素，中和SARS-CoV-2	[87]
		N-叠氮基乙酰基甘露糖胺 ManNAz	插入N₃偶联抗 CD3ε抗体，激活肿瘤免疫	[88]
			插入N₃，增强细胞膜-纳米核结合	[89]
			插入N₃偶联ALN，靶向骨组织	[90]
	GalNAc补救合成途径	N-乙酰半乳糖胺 GalNAc	插入N₃，靶向肿瘤细胞BCN	[91]
脂质工程	CDP-胆碱途径	胆碱 Choline	插入N₃偶联RGD，靶向肿瘤	[35]
			插入N₃偶联pMHC-I/CD28，激活肿瘤免疫	[92]
			插入N₃偶联抗CD205，激活肿瘤免疫	[93]

Fig. 4 Schematic illustration of N3-labeled T cell membrane-biomimetic nanoparticles with dual-targeting mechanism [91][A) Synthesis of N3-TINPs. B) Tumor cells carrying the BCN group via natural glycometabolic labeling by pretreatment with Ac4ManN-BCN. N3-TINPs could target tumor through immune recognition of T cell membrane and bioorthogonal reaction between BCN and N3 groups.]

Fig. 5 Additional functions of SCNPs enabled by synthetic biology. Created in BioRender. Huang Yang (2025).

Table 3 The application of SCNPs in disease diagnosis and treatment

应用	功能	机制	参考文献
恶性肿瘤	肿瘤靶向	修饰靶向分子，如RGD、NGR	[110,35]
	免疫疗法	表达免疫检测点抗体或受体，阻断PD-1/PD-L1 信号轴；	[61,75]
	免疫疗法	修饰肿瘤抗原和/或肿瘤细胞膜固有抗原激活肿瘤免疫，发挥疫苗功效	[79]
心血管疾病	炎症靶向和缓解	修饰靶向蛋白如VLA-4；血小板、巨噬细胞等固有炎症靶向和免疫调节能力	[111,112,81]
感染性疾病	中和病原体	修饰对应受体/配体/抗体，如肝素	[87,106]
感染性疾病	激活免疫应答	展示病毒特异性抗原表位	[113]
自身免疫性疾病	抑制免疫激活	修饰对应受体，如OX40、CD40，阻断免疫激活信号	[114]
神经退行性疾病	跨越血脑屏障实现靶向递送	神经干细胞脑归巢效应；修饰靶向分子，如CCR2	[78,115]
骨相关疾病	骨靶向	修饰靶向分子，如CXCR4、ALN	[90,116]
骨相关疾病	中和破骨因子	过表达对应受体，如RANK	[117]

Fig. 6 Schematic Illustration of Preparing Engineered Macrophage-Membrane-Coated Nanoparticles with Enhanced PD-1 Expression[61][Macrophage membrane-derived NPs permeate the blood-brain-barrier and conduct targeted drug delivery; in situ engineered overexpression of PD-1 on the macrophage membrane competitively binds to PD-L1 and effectively blocks the PD-1/PD-L1 signaling axis.]

Fig.7 Schematic representation of MMM/RNA NPs-mediated siRNA delivery for the treatment of MIRI[81][A) Depiction of the MMM/RNA NPs preparation. Macrophages were transfected with adenoviruses encoding for HA and RAGE to construct engineered macrophages. B) Diagram of MMM/RNA NPs injected into the tail vein of myocardial ischemia-reperfusion mice. Myocardial ischemic injury recruits a large number of neutrophils in the blood, which activate and release S100A9 inflammatory factors. MMM/RNA NPs rely on their cell membrane protein RAGE to recruit to the myocardial injury area along the concentration of S100A9. Neutrophils engulf MMM/RNA NPs, and MMM/RNA NPs rely on their cell membrane protein HA to play an endosomal escape role, transporting siRNA to the cytoplasm.]

Fig. 8 Schematic illustration of nanosponges against COVID-19[106][A) Preparation of nanosponges by fusing cellular membrane nanovesicles derived from genetically edited 293T/ACE2 and THP-1 cells. The nanosponges, displaying abundant ACE2 and cytokine receptors, compete with host cells to bind (B) SARS-CoV-2 and (C) inflammatory cytokines, such as IL-6 and GM-CSF.]

Fig. 9 The scale production process of SCNPs. Created in BioRender. Huang Yang (2025).

References 116

[117]	Zhou Y., Deng Y. K., Liu Z. M., et al. Cytokine-scavenging nanodecoys reconstruct osteoclast/osteoblast balance toward the treatment of postmenopausal osteoporosis [J]. Science Advances, 2021, 7(48): eabl6432.
[118]	Jiang X. B., Zhang X. Y., Guo C., et al. Genetically engineered cell membrane-coated magnetic nanoparticles for high-performance isolation of circulating tumor cells [J]. Advanced Functional Materials, 2024, 34(7): 2304426.
[119]	Song Y. N., Zhang N., Li Q. Y., et al. Biomimetic liposomes hybrid with platelet membranes for targeted therapy of atherosclerosis [J]. Chemical Engineering Journal, 2021, 408: 127296.
[120]	Su T., Huang K., Ma H., et al. Platelet-inspired nanocells for targeted heart repair after ischemia/reperfusion injury [J]. Advanced Functional Materials, 2019, 29(4): 1803567.
[121]	Pang X., Liu X., Cheng Y., et al. Sono-immunotherapeutic nanocapturer to combat multidrug-resistant bacterial infections [J]. Advanced Materials, 2019, 31(35): 1902530.
[122]	Fang T. L., Li B. Q., Li M., et al. Engineered cell membrane vesicles expressing cd40 alleviate system lupus nephritis by intervening b cell activation [J]. Small Methods, 2023, 7(3): 2200925.
[123]	Anzalone A. V., Randolph P. B., Davis J. R., et al. Search-and-replace genome editing without double-strand breaks or donor DNA [J]. Nature, 2019, 576(7785): 149-157.
[1]	Beach M. A., Nayanathara U., Gao Y. T., et al. Polymeric nanoparticles for drug delivery [J]. Chemical Reviews, 2024, 124(9): 5505-5616.
[2]	Dams E. T. M., Laverman P., Oyen W. J. G., et al. Accelerated blood clearance and altered biodistribution of repeated injections of sterically stabilized liposomes [J]. Journal of Pharmacology and Experimental Therapeutics, 2000, 292(3): 1071-1079.
[3]	Mohamed M., Abu Lila A. S., Shimizu T., et al. Pegylated liposomes: Immunological responses [J]. Science and Technology of Advanced Materials, 2019, 20(1): 710-724.
[4]	Sanna V., Pala N., Sechi M.. Targeted therapy using nanotechnology: Focus on cancer [J]. International Journal of Nanomedicine, 2014, 9(1): 467-483.
[5]	An Y. Q., Ji C., Zhang H., et al. Engineered cell membrane coating technologies for biomedical applications: From nanoscale to macroscale [J]. Acs Nano, 2025, 19(12): 11517-11546.
[6]	Le Q. V., Lee J., Lee H., et al. Cell membrane-derived vesicles for delivery of therapeutic agents [J]. Acta Pharmaceutica Sinica B, 2021, 11(8): 2096-2113.
[7]	Hu C. M. J., Zhang L., Aryal S., et al. Erythrocyte membrane-camouflaged polymeric nanoparticles as a biomimetic delivery platform [J]. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(27): 10980-10985.
[8]	Parodi A., Quattrocchi N., Van De Ven A. L., et al. Synthetic nanoparticles functionalized with biomimetic leukocyte membranes possess cell-like functions [J]. Nature Nanotechnology, 2013, 8(1): 61-68.
[9]	Cao B. R., Yang M. Y., Zhu Y., et al. Stem cells loaded with nanoparticles as a drug carrier for in vivo breast cancer therapy [J]. Advanced Materials, 2014, 26(27): 4627-4631.
[10]	Deng G. J., Sun Z. H., Li S. P., et al. Cell-membrane immunotherapy based on natural killer cell membrane coated nanoparticles for the effective inhibition of primary and abscopal tumor growth [J]. Acs Nano, 2018, 12(12): 12096-12108.
[11]	Fang R. H., Hu C. M. J., Luk B. T., et al. Cancer cell membrane-coated nanoparticles for anticancer vaccination and drug delivery [J]. Nano Letters, 2014, 14(4): 2181-2188.
[12]	Gong H., Zhang Q. Z., Komarla A., et al. Nanomaterial biointerfacing via mitochondrial membrane coating for targeted detoxification and molecular detection [J]. Nano Letters, 2021, 21(6): 2603-2609.
[13]	Jiang L. X., Zhu Y. Y., Luan P. W., et al. Bacteria-anchoring hybrid liposome capable of absorbing multiple toxins for antivirulence therapy of escherichia coli infection [J]. Acs Nano, 2021, 15(3): 4173-4185.
[14]	Xuan M. J., Shao J. X., Dai L. R., et al. Macrophage cell membrane camouflaged mesoporous silica nanocapsules for in vivo cancer therapy [J]. Advanced Healthcare Materials, 2015, 4(11): 1645-1652.
[15]	Hu C. M. J., Fang R. H., Copp J., et al. A biomimetic nanosponge that absorbs pore-forming toxins [J]. Nature Nanotechnology, 2013, 8(5): 336-340.
[16]	Yang L., Froio R. M., Sciuto T. E., et al. Icam-1 regulates neutrophil adhesion and transcellular migration of tnf-α-activated vascular endothelium under flow [J]. Blood, 2005, 106(2): 584-592.
[17]	Zhou K., Yang C. L., Shi K., et al. Activated macrophage membrane-coated nanoparticles relieve osteoarthritis-induced synovitis and joint damage [J]. Biomaterials, 2023, 295: 122036.
[18]	Guo J., Pan X. T., Wu Q. Y., et al. Bio-barrier-adaptable biomimetic nanomedicines combined with ultrasound for enhanced cancer therapy [J]. Signal Transduction and Targeted Therapy, 2025, 10(1): 137.
[19]	Lei S. B., Li J. M., Zhu M. F., et al. Chimeric antigen receptor-engineered cell membrane-coated nanoparticles promote dual-targeted mrna-based cancer gene therapy [J]. Acs Nano, 2025, 19(16): 15668-15684.
[20]	Liu H., Su Y. Y., Jiang X. C., et al. Cell membrane-coated nanoparticles: A novel multifunctional biomimetic drug delivery system [J]. Drug Delivery and Translational Research, 2023, 13(3): 716-737.
[21]	Tuo Z., He Q. Y., Zhang Z. J., et al. Irradiation conditioning of adjuvanted, autologous cancer cell membrane nanoparticle vaccines [J]. Chemical Engineering Journal, 2022, 433: 134437.
[22]	Wang X. X., Zhu X. Q., Li B. Y., et al. Intelligent biomimetic nanoplatform for systemic treatment of metastatic triple-negative breast cancer via enhanced egfr-targeted therapy and immunotherapy [J]. Acs Applied Materials & Interfaces, 2022, 14(20): 23152-23163.
[23]	Zhu J. Y., Zheng D. W., Zhang M. K., et al. Preferential cancer cell self-recognition and tumor self-targeting by coating nanoparticles with homotypic cancer cell membranes [J]. Nano Letters, 2016, 16(9): 5895-5901.
[24]	Wang S. Y., Wang D., Kai M. X., et al. Design strategies for cellular nanosponges as medical countermeasures [J]. Bme Frontiers, 2023, 4: 0018.
[25]	Zhang J. A., Feng K. L., Shen W. T., et al. Research advances of cellular nanoparticles as multiplex countermeasures [J]. Acs Nano, 2024, 18(44): 30211-30223.
[26]	Pang Z. Q., Hu C. M. J., Fang R. H., et al. Detoxification of organophosphate poisoning using nanoparticle bioscavengers [J]. Acs Nano, 2015, 9(6): 6450-6458.
[27]	Ai X. Z., Wang S. Y., Duan Y. O., et al. Emerging approaches to functionalizing cell membrane-coated nanoparticles [J]. Biochemistry, 2021, 60(13): 941-955.
[28]	Fang R. H., Gao W. W., Zhang L. F.. Targeting drugs to tumours using cell membrane-coated nanoparticles [J]. Nature Reviews Clinical Oncology, 2023, 20(1): 33-48.
[29]	Yi W. Z., Xiao P., Liu X. C., et al. Recent advances in developing active targeting and multi-functional drug delivery systems via bioorthogonal chemistry [J]. Signal Transduction and Targeted Therapy, 2022, 7(1): 386.
[30]	Zhang X. E., Liu C. L., Dai J. B., et al. Enabling technology and core theory of synthetic biology [J]. Science China-Life Sciences, 2023, 66(8): 1742-1785.
[31]	Jiang Y., Krishnan N., Zhou J. R., et al. Engineered cell-membrane-coated nanoparticles directly present tumor antigens to promote anticancer immunity [J]. Advanced Materials, 2020, 32(30): 2001808.
[32]	Krishnan N., Jiang Y., Zhou J. R., et al. A modular approach to enhancing cell membrane-coated nanoparticle functionality using genetic engineering [J]. Nature Nanotechnology, 2024, 19(3): 345–353.
[33]	Ma W. J., Zhu D. M., Li J. H., et al. Coating biomimetic nanoparticles with chimeric antigen receptor t cell-membrane provides high specificity for hepatocellular carcinoma photothermal therapy treatment [J]. Theranostics, 2020, 10(3): 1281-1295.
[34]	Ai X. Z., Wang D., Honko A., et al. Surface glycan modification of cellular nanosponges to promote sars-cov-2 inhibition [J]. Journal of the American Chemical Society, 2021, 143(42): 17615-17621.
[35]	Zhang F., Zhao L. J., Wang S. M., et al. Construction of a biomimetic magnetosome and its application as a sirna carrier for high-performance anticancer therapy [J]. Advanced Functional Materials, 2018, 28(1): 1703326.
[36]	Chugh V., Krishna K. V., Pandit A.. Cell membrane-coated mimics: A methodological approach for fabrication, characterization for therapeutic applications, and challenges for clinical translation [J]. Acs Nano, 2021, 15(11): 17080-17123.
[37]	Wang Y., Zhang K., Qin X., et al. Biomimetic nanotherapies: Red blood cell based core-shell structured nanocomplexes for atherosclerosis management [J]. Advanced Science, 2019, 6(12): 1900172.
[38]	Wei X. L., Gao J., Fang R. H., et al. Nanoparticles camouflaged in platelet membrane coating as an antibody decoy for the treatment of immune thrombocytopenia [J]. Biomaterials, 2016, 111: 116-123.
[39]	Park J. H., Jiang Y., Zhou J. R., et al. Genetically engineered cell membrane-coated nanoparticles for targeted delivery of dexamethasone to inflamed lungs [J]. Science Advances, 2021, 7(25): eabf7820.
[40]	He Y. L., Zhang S. Q., She Y. G., et al. Innovative utilization of cell membrane-coated nanoparticles in precision cancer therapy [J]. Exploration, 2024, 4(6): 20230164.
[41]	Rao L., Cai B., Bu L. L., et al. Microfluidic electroporation-facilitated synthesis of erythrocyte membrane-coated magnetic nanoparticles for enhanced imaging-guided cancer therapy [J]. Acs Nano, 2017, 11(4): 3496-3505.
[42]	Hu C. M. J., Fang R. H., Wang K. C., et al. Nanoparticle biointerfacing by platelet membrane cloaking [J]. Nature, 2015, 526(7571): 118–121.
[43]	Sekhon U. D. S., Swingle K., Girish A., et al. Platelet-mimicking procoagulant nanoparticles augment hemostasis in animal models of bleeding [J]. Science Translational Medicine, 2022, 14(629): eabb8975.
[44]	Lavergne M., Janus-Bell E., Schaff M., et al. Platelet integrins in tumor metastasis: Do they represent a therapeutic target? [J]. Cancers, 2017, 9(10): 133.
[45]	Chen L., Zhou Z. Y., Hu C., et al. Platelet membrane-coated nanocarriers targeting plaques to deliver anti-cd47 antibody for atherosclerotic therapy [J]. Research, 2022, 2022: 9845459.
[46]	Li Y., Xiang Q., Zhang Y., et al. Mechanical biomimetic nanocomposites for multidimensional treatment of arterial thrombosis [J]. Advanced Science, 2025, 12(25): 2501134.
[47]	Song Y. J., He Y. H., Rong L., et al. "Platelet-coated bullets" biomimetic nanoparticles to ameliorate experimental colitis by targeting endothelial cells [J]. Biomaterials Advances, 2023, 148: 213378.
[48]	Tao W. H., Zhao D. Y., Li G. T., et al. Artificial tumor microenvironment regulated by first hemorrhage for enhanced tumor targeting and then occlusion for synergistic bioactivation of hypoxia-sensitive platesomes [J]. Acta Pharmaceutica Sinica B, 2022, 12(3): 1487-1499.
[49]	Gao C., Huang Q. X., Liu C. H., et al. Treatment of atherosclerosis by macrophage-biomimetic nanoparticles via targeted pharmacotherapy and sequestration of proinflammatory cytokines [J]. Nature Communications, 2020, 11(1): 2622.
[50]	Song W. L., Jia P. F., Ren Y. P., et al. Engineering white blood cell membrane-camouflaged nanocarriers for inflammation-related therapeutics [J]. Bioactive Materials, 2023, 23: 80-100.
[51]	Wu Y. S., Wan S. L., Yang S., et al. Macrophage cell membrane-based nanoparticles: A new promising biomimetic platform for targeted delivery and treatment [J]. Journal of Nanobiotechnology, 2022, 20(1): 542.
[52]	Zhu K. H., Xu Y. C., Zhong R., et al. Hybrid liposome-erythrocyte drug delivery system for tumor therapy with enhanced targeting and blood circulation [J]. Regenerative Biomaterials, 2023, 10: rbad045.
[53]	Ren E., Liu C., Lv P., et al. Genetically engineered cellular membrane vesicles as tailorable shells for therapeutics [J]. Advanced Science, 2021, 8(21): 2100460.
[54]	Mcmahon H. T., Gallop J. L.. Membrane curvature and mechanisms of dynamic cell membrane remodelling [J]. Nature, 2005, 438(7068): 590-596.
[55]	Owji H., Nezafat N., Negandaripour M., et al. A comprehensive review of signal peptides: Structure, roles, and applications [J]. European Journal of Cell Biology, 2018, 97(6): 422-441.
[56]	Voorhees R. M., Hegde R. S.. Toward a structural understanding of co-translational protein translocation [J]. Current Opinion in Cell Biology, 2016, 41: 91-99.
[57]	Jiang R. Z., Yuan S. T., Zhou Y. L., et al. Strategies to overcome the challenges of low or no expression of heterologous proteins in escherichia coli [J]. Biotechnology Advances, 2024, 75: 108417.
[58]	Manandhar M., Chun E., Romesberg F. E.. Genetic code expansion: Inception, development, commercialization [J]. Journal of the American Chemical Society, 2021, 143(13): 4859-4878.
[59]	Parvathy S. T., Udayasuriyan V., Bhadana V.. Codon usage bias [J]. Molecular Biology Reports, 2022, 49(1): 539-565.
[60]	Guo X., Ye S. P., Cheng X. Y., et al. Engineered P2Y₁₂-overexpressing cell-membrane-wrapped nanoparticles for the functional reversal of ticagrelor and clopidogrel [J]. Nano Letters, 2024, 24(34): 10482-10489.
[61]	Yin T. Y., Fan Q., Hu F. F., et al. Engineered macrophage-membrane-coated nanoparticles with enhanced pd-1 expression induce immunomodulation for a synergistic and targeted antiglioblastoma activity [J]. Nano Letters, 2022, 22(16): 6606-6614.
[62]	Krishnan N., Peng F. X., Mohapatra A., et al. Genetically engineered cellular nanoparticles for biomedical applications [J]. Biomaterials, 2023, 296: 122065.
[63]	Sayed N., Allawadhi P., Khurana A., et al. Gene therapy: Comprehensive overview and therapeutic applications [J]. Life Sciences, 2022, 294: 120375.
[64]	Stewart M. P., Langer R., Jensen K. F.. Intracellular delivery by membrane disruption: Mechanisms, strategies, and concepts [J]. Chemical Reviews, 2018, 118(16): 7409-7531.
[65]	Shi J. F., Ma Y. F., Zhu J., et al. A review on electroporation-based intracellular delivery [J]. Molecules, 2018, 23(11): 3044.
[66]	Tsong T. Y.. Electroporation of cell-membranes [J]. Biophysical Journal, 1991, 60(2): 297-306.
[67]	Chong Z. X., Yeap S. K., Ho W. Y.. Transfection types, methods and strategies: A technical review [J]. Peerj, 2021, 9: e11165.
[68]	Meng L., Liu X. F., Wang Y. C., et al. Sonoporation of cells by a parallel stable cavitation microbubble array [J]. Advanced Science, 2019, 6(17): 1900557.
[69]	Pylaev T., Vanzha E., Avdeeva E., et al. A novel cell transfection platform based on laser optoporation mediated by au nanostar layers [J]. Journal of Biophotonics, 2019, 12(1): e201800166.
[70]	Chow Y. T., Chen S. X., Wang R., et al. Single cell transfection through precise microinjection with quantitatively controlled injection volumes [J]. Scientific Reports, 2016, 6: 24127.
[71]	Zhang J. Q., Hu Y. X., Yang J. X., et al. Non-viral, specifically targeted car-t cells achieve high safety and efficacy in b-nhl [J]. Nature, 2022, 609(7926): 369-374.
[72]	Kiefer K., Clement J., Garidel P., et al. Transfection efficiency and cytotoxicity of nonviral gene transfer reagents in human smooth muscle and endothelial cells [J]. Pharmaceutical Research, 2004, 21(6): 1009-1017.
[73]	Kim H. Y., Kang M., Choo Y. W., et al. Immunomodulatory lipocomplex functionalized with photosensitizer-embedded cancer cell membrane inhibits tumor growth and metastasis [J]. Nano Letters, 2019, 19(8): 5185-5193.
[74]	Liu S. Y., Wu J. Y., Feng Y. J., et al. Cd47ko/crt dual-bioengineered cell membrane-coated nanovaccine combined with anti-pd-l1 antibody for boosting tumor immunotherapy [J]. Bioactive Materials, 2023, 22: 211-224.
[75]	Zhang X. D., Wang C., Wang J. Q., et al. Pd-1 blockade cellular vesicles for cancer immunotherapy [J]. Advanced Materials, 2018, 30(22): 1707112.
[76]	Bose R. J. C., Kim B. J., Arai Y., et al. Bioengineered stem cell membrane functionalized nanocarriers for therapeutic targeting of severe hindlimb ischemia [J]. Biomaterials, 2018, 185: 360-370.
[77]	Gu C. J., Geng X. W., Wu Y. C., et al. Engineered macrophage membrane-coated nanoparticles with enhanced ccr2 expression promote spinal cord injury repair by suppressing neuroinflammation and neuronal death [J]. Small, 2024, 20(10): 2305659.
[78]	Huang D. H., Wang Q. W., Cao Y. H., et al. Multiscale nir-ii imaging-guided brain-targeted drug delivery using engineered cell membrane nanoformulation for alzheimer's disease therapy [J]. Acs Nano, 2023, 17(5): 5033-5046.
[79]	Liu C., Liu X., Xiang X. C., et al. A nanovaccine for antigen self-presentation and immunosuppression reversal as a personalized cancer immunotherapy strategy [J]. Nature Nanotechnology, 2022, 17(5): 531-540.
[80]	Liu Z. M., Zhou X. F., Li Q., et al. Macrophage-evading and tumor-specific apoptosis inducing nanoparticles for targeted cancer therapy [J]. Acta Pharmaceutica Sinica B, 2023, 13(1): 327-343.
[81]	Lu H., Wang J. Z., Chen Z. W., et al. Engineered macrophage membrane-coated s100a9-sirna for ameliorating myocardial ischemia-reperfusion injury [J]. Advanced Science, 2024, 11(41): 2403542.
[82]	Ma J. N., Zhang S. Q., Liu J., et al. Targeted drug delivery to stroke via chemotactic recruitment of nanoparticles coated with membrane of engineered neural stem cells [J]. Small, 2019, 15(35): 1902011.
[83]	Bulcha J. T., Wang Y., Ma H., et al. Viral vector platforms within the gene therapy landscape [J]. Signal Transduction and Targeted Therapy, 2021, 6(1): 53.
[84]	Crystal R. G.. Adenovirus: The first effective in vivo gene delivery vector [J]. Human Gene Therapy, 2014, 25(1): 3-11.
[85]	Milone M. C., O'doherty U.. Clinical use of lentiviral vectors [J]. Leukemia, 2018, 32(7): 1529-1541.
[86]	Park J. H., Mohapatra A., Zhou J. R., et al. Virus-mimicking cell membrane-coated nanoparticles for cytosolic delivery of mrna [J]. Angewandte Chemie-International Edition, 2022, 61(2): e202113671.
[87]	Ai X. Z., Wang D., Noh I., et al. Glycan-modified cellular nanosponges for enhanced neutralization of botulinum toxin [J]. Biomaterials, 2023, 302.
[88]	Xiao P., Wang J., Zhao Z. T., et al. Engineering nanoscale artificial antigen-presenting cells by metabolic dendritic cell labeling to potentiate cancer immunotherapy [J]. Nano Letters, 2021, 21(5): 2094-2103.
[89]	Zhang X. L., Zhen X. Y., Yang Y. X., et al. Precise assembly of inside-out cell membrane camouflaged nanoparticles via bioorthogonal reactions for improving drug leads capturing [J]. Acta Pharmaceutica Sinica B, 2023, 13(2): 852-862.
[90]	Huang Y., Chen M. W., Shen Y. D., et al. Bone-targeting cell membrane-engineered caco3-based nanoparticles restore local bone homeostasis for microenvironment-responsive osteoporosis treatment [J]. Chemical Engineering Journal, 2023, 470: 144145.
[91]	Han Y. T., Pan H., Li W. J., et al. T cell membrane mimicking nanoparticles with bioorthogonal targeting and immune recognition for enhanced photothermal therapy [J]. Advanced Science, 2019, 6(15): 1900251.
[92]	Zhang Q. B., Wei W., Wang P. L., et al. Biomimetic magnetosomes as versatile artificial antigen-presenting cells to potentiate t-cell-based anticancer therapy [J]. Acs Nano, 2017, 11(11): 10724-10732.
[93]	Li F., Nie W. D., Zhang F., et al. Engineering magnetosomes for high-performance cancer vaccination [J]. Acs Central Science, 2019, 5(5): 796-807.
[94]	Yoon H. Y., Koo H., Kim K., et al. Molecular imaging based on metabolic glycoengineering and bioorthogonal click chemistry [J]. Biomaterials, 2017, 132: 28-36.
[95]	Krishnan N., Fang R. N. H., Zhang L. F.. Engineering of stimuli-responsive self-assembled biomimetic nanoparticles [J]. Advanced Drug Delivery Reviews, 2021, 179: 114006.
[96]	Boussiotis V. A.. Molecular and biochemical aspects of the pd-1 checkpoint pathway [J]. New England Journal of Medicine, 2016, 375(18): 1767-1778.
[97]	Tang Q. S., Sun S. H., Wang P., et al. Genetically engineering cell membrane-coated bto nanoparticles for mmp2-activated piezocatalysis-immunotherapy [J]. Advanced Materials, 2023, 35(18): 2300964.
[98]	Xu S. Y., Shi X. X., Ren E., et al. Genetically engineered nanohyaluronidase vesicles: A smart sonotheranostic platform for enhancing cargo penetration of solid tumors [J]. Advanced Functional Materials, 2022, 32(22): 2112989.
[99]	Han J. H., Sul J. H., Lee J. M., et al. Engineered exosomes with a photoinducible protein delivery system enable crispr-cas-based epigenome editing in alzheimer's disease [J]. Science Translational Medicine, 2024, 16(759): eadi4830.
[100]	Yang Y. X., Wang K., Pan Y. W., et al. Engineered cell membrane-derived nanoparticles in immune modulation [J]. Advanced Science, 2021, 8(24): 2102330.
[101]	Zhou J. R., Kroll A. V., Holay M., et al. Biomimetic nanotechnology toward personalized vaccines [J]. Advanced Materials, 2020, 32(13): 1901255.
[102]	Wang K. Y., Zhang X. B., Ye H., et al. Biomimetic nanovaccine-mediated multivalent il-15 self-transpresentation (mist) for potent and safe cancer immunotherapy [J]. Nature Communications, 2023, 14(1): 6748.
[103]	Yi M., Zheng X. L., Niu M. K., et al. Combination strategies with pd-1/pd-l1 blockade: Current advances and future directions [J]. Molecular Cancer, 2022, 21(1): 28.
[104]	Ding L., Zhang X. L., Yu P. W., et al. Genetically engineered nanovesicles mobilize synergistic antitumor immunity by adar1 silence and pdl1 blockade [J]. Molecular Therapy, 2023, 31(8): 2489-2506.
[105]	Wang S. Y., Wang D., Duan Y. O., et al. Cellular nanosponges for biological neutralization [J]. Advanced Materials, 2022, 34(13): 2107719.
[106]	Rao L., Xia S., Xu W., et al. Decoy nanoparticles protect against covid-19 by concurrently adsorbing viruses and inflammatory cytokines [J]. Proceedings of the National Academy of Sciences of the United States of America, 2020, 117(44): 27141-27147.
[107]	Schlapschy M., Binder U., Börger C., et al. Pasylation: A biological alternative to pegylation for extending the plasma half-life of pharmaceutically active proteins [J]. Protein Engineering Design & Selection, 2013, 26(8): 489-501.
[108]	Duan Y. U., Zhou J. R., Zhou Z. D., et al. Extending the in vivo residence time of macrophage membrane-coated nanoparticles through genetic modification [J]. Small, 2023, 19(52): 2305551.
[109]	Krishnamurthy S., Muthukumaran P., Jayakumar M. K. G., et al. Surface protein engineering increases the circulation time of a cell membrane-based nanotherapeutic [J]. Nanomedicine-Nanotechnology Biology and Medicine, 2019, 18: 169-178.
[110]	Lv P., Liu X., Chen X. M., et al. Genetically engineered cell membrane nanovesicles for oncolytic adenovirus delivery: A versatile platform for cancer virotherapy [J]. Nano Letters, 2019, 19(5): 2993-3001.
[111]	Chen T. Y., Lv W. T., Yang M. X., et al. Treatment of atherosclerosis by biomimetic responsive-targeting hybrid nanoparticles enabled by genetically engineering cell membranes and nucleic acid nanostructures for atorvastatin delivery [J]. Chemical Engineering Journal, 2025, 511: 162130.
[112]	Li Y. Y., Che J. Y., Chang L., et al. Cd47-and integrin α4/β1-comodified-macrophage-membrane-coated nanoparticles enable delivery of colchicine to atherosclerotic plaque [J]. Advanced Healthcare Materials, 2022, 11(4): 2101788.
[113]	Zhang P. F., Chen Y. X., Zeng Y., et al. Virus-mimetic nanovesicles as a versatile antigen-delivery system [J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(45): E6129-E6138.
[114]	Fu Y., Wang L. L., Liu W., et al. Ox40l blockade cellular nanovesicles for autoimmune diseases therapy [J]. Journal of Controlled Release, 2021, 337: 557-570.
[115]	Lin R. R., Jin L. L., Xue Y. Y., et al. Hybrid membrane-coated nanoparticles for precise targeting and synergistic therapy in alzheimer's disease [J]. Advanced Science, 2024, 11(24): 2306675.
[116]	Liu H., Song P. R., Zhang H., et al. Synthetic biology-based bacterial extracellular vesicles displaying bmp-2 and cxcr4 to ameliorate osteoporosis [J]. Journal of Extracellular Vesicles, 2024, 13(4): e12429.