Antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system

Abstract

This invention discloses a PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNPs, belonging to the field of biomedical technology. It comprises: an LNP, mRNA encoding the PIEZO2 protein encapsulated within the LNP, and a CD31 antibody conjugated to the surface of the LNP. By combining cationic lipid (DOTAP)-mediated lung tissue targeting and CD31 antibody-mediated vascular endothelial cell targeting, dual precision delivery at the organ-to-cell level is achieved, greatly improving the accuracy and efficiency of treatment and reducing off-target effects.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP. Background Technology

[0002] Acute lung injury (ALI) and its severe form, acute respiratory distress syndrome (ARDS), are common critical illnesses with high mortality rates, and currently lack specific treatments. Pulmonary vascular endothelial cell injury is one of the core pathological processes in ALI / ARDS. Studies have shown that the mechanosensitive ion channel PIEZO2 in endothelial cells plays a crucial role in maintaining endothelial barrier function and suppressing inflammatory responses. In various lung injury models, the expression of endothelial cell PIEZO2 is significantly downregulated, and its impaired function exacerbates vascular leakage and inflammatory cell infiltration.

[0003] Messenger RNA (mRNA) therapy offers a novel strategy for compensating for the loss of functional proteins. However, achieving organ- and cell-specific delivery of mRNA remains a major challenge. Lipid nanoparticles (LNPs) are currently the most advanced mRNA delivery vectors, but they naturally tend to be taken up by the liver. Existing LNP lung-targeting strategies (such as adjusting the lipid composition to be positively charged) can improve lung tissue enrichment, but cell targeting remains insufficient, making it difficult to specifically target the key cells in disease—vascular endothelial cells.

[0004] Therefore, developing a system that can efficiently and specifically deliver therapeutic mRNA (such as PIEZO2 mRNA) to pulmonary vascular endothelial cells is of great significance for the treatment of lung injury. Summary of the Invention

[0005] To address the shortcomings of existing technologies, the present invention aims to provide a PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP, thus solving the problems in the prior art.

[0006] The objective of this invention can be achieved through the following technical solutions: The antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system includes: LNP, mRNA encoding PIEZO2 protein encapsulated within LNP, and CD31 antibody conjugated to the surface of LNP.

[0007] Furthermore, the LNP comprises the following molar percentages of lipid components: Ionizable cationic lipids: 30%; Cationic lipids: 40%; PEGylated lipids: 2%; DSPC: 6%; Cholesterol: 22%.

[0008] Furthermore, the ionizable cationic lipid is SM-102.

[0009] Furthermore, the cationic lipid is DOTAP.

[0010] Further, the PEGylated lipids include: neutral PEGylated lipids and DSPE-PEG2000-Mal; the neutral PEGylated lipids are DMG-PEG2000 or DSPE-PEG.

[0011] Furthermore, the molar percentage of DSPE-PEG2000-Mal is 0.5% of LNP.

[0012] Furthermore, the CD31 antibody is covalently coupled to the maleimide group at the end of DSPE-PEG-Mal via a thiol group.

[0013] The preparation method of the above-mentioned antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system includes the following steps: S1, dissolve ionizable cationic lipids, cationic lipids and PEGylated lipids in an organic solvent, remove the solvent, and form a lipid film; S2, the lipid membrane was hydrated with acidic buffer solution and then homogenized to form a blank LNP; S3, encapsulate the mRNA encoding PIEZO2 inside the LNP; S4. After purifying the LNP encapsulated with PIEZO2 mRNA, it is mixed with CD31 antibody treated with reducing agent to obtain antibody-conjugated LNP-PIEZO2 mRNA complex, i.e., delivery system. The above-mentioned antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system is used in the preparation of drugs for the treatment or prevention of lung injury.

[0014] A drug for treating or preventing lung injury, comprising: the above-described antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system.

[0015] The beneficial effects of this invention are: 1. This invention combines DOTAP-mediated lung tissue targeting and CD31 antibody-mediated vascular endothelial cell targeting, achieving dual precision delivery from the organ to the cell level, which greatly improves the accuracy and efficiency of treatment and reduces off-target effects.

[0016] 2. This invention targets the core pathological mechanism of PIEZO2 downregulation in endothelial cells during lung injury. It directly supplements functional proteins through mRNA technology, intervening in the disease process at the etiological level. The theory is novel and the strategy is advanced.

[0017] 3. By carefully optimizing the LNP components (such as controlling the total PEG content and the ratio of DSPE-PEG2000-Mal), this invention introduces antibody function while maximizing the preservation of LNP stability and biocompatibility, and avoids loss of targeting due to excessive PEG shielding.

[0018] 4. Through in vitro and in vivo experiments, this invention has demonstrated that the delivery system can significantly improve the transfection efficiency of PIEZO2 mRNA in pulmonary vascular endothelial cells and effectively upregulate PIEZO2 protein expression, thereby significantly reducing pulmonary vascular leakage, inflammatory response and tissue damage in model animals. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 This is a schematic diagram of the delivery system of the present invention; Figure 2 The figures show the particle size distribution of the LNPs prepared in the examples and the statistical results of the Zeta potential characterization of the LNPs. Figure 3 This is an in vitro experimental diagram; Figure 4 This is an in vivo distribution map of a mouse lung injury model; Figure 5 These are images of mouse lung tissue pathological sections (H&E staining), electron microscopy, and Western blotting results. Figure 6 This is a colocalization analysis diagram of immunofluorescence staining on mouse lung tissue sections; Figure 7 This is a graph showing the pharmacodynamic evaluation results of the CD31-targeted LNP delivery system in treating acute lung injury in mice. Detailed Implementation

[0021] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0022] like Figure 1 As shown, the PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP includes: LNP, mRNA encoding PIEZO2 protein encapsulated in LNP, and CD31 antibody (PECAM-1 antibody) conjugated on the surface of LNP. LNP comprises the following molar percentages of lipid components: Ionizable cationic lipids: 30%; Cationic lipids: 40%; DSPC (1,2-distearyl-sn-glycerol-3-phosphocholine): 6%; Cholesterol: 22%; PEGylated lipids: 2%; The ionizable cationic lipid is SM-102; The cationic lipid is DOTAP; The PEGylated lipids include: neutral PEGylated lipids (such as DMG-PEG2000 or DSPE-PEG) and functional PEGylated lipids (DSPE-PEG2000-Mal), and the molar percentage of the functional PEGylated lipids is 0.5% of the total lipids (LNP).

[0023] The CD31 antibody is covalently coupled to the thiol (-SH) group of the CD31 antibody via the maleimide group at the end of the DSPE-PEG-Mal.

[0024] The mRNA encoding the PIEZO2 protein was specifically derived from: Coste B, Mathur J, Schmidt M, Earley TJ, Ranade S, Petrus MJ, Dubin AE, Patapoutian A. Piezo1 and Piezo2 are essential components of distinct mechanically activated cation channels. Science. 2010 Oct 1;330(6000):55-60. doi: 10.1126 / science.1193270. Epub 2010Sep 2. PMID: 20813920; PMCID: PMC3062430. The preparation steps of the antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system include: S1, Dissolve ionizable cationic lipids, cationic lipids, and PEGylated lipids in an organic solvent, remove the solvent, and form a lipid film. S2, the lipid film was hydrated with an acidic buffer (citrate buffer, pH 4.0) and then homogenized by an extruder to form a blank LNP; S3, using a remote loading method, encapsulates the mRNA encoding PIEZO2 (PIEZO2 mRNA for short) inside the LNP; S4. After purifying the LNP containing PIEZO2 mRNA, it is mixed with CD31 antibody treated with a reducing agent (to expose thiol groups) under mild conditions to obtain the antibody-conjugated LNP-PIEZO2 mRNA complex (i.e., delivery system). The aforementioned delivery system can be used to prepare drugs for the prevention or treatment of lung injury, wherein the lung injury includes, but is not limited to: lung injury caused by sepsis, acid inhalation lung injury, ventilator-associated lung injury, and lung injury caused by bacterial or viral pneumonia.

[0025] Example 1 This embodiment describes a specific delivery system (structural diagram as shown in the figure). Figure 1 (as shown) and its preparation process; LNP includes a molar percentage of lipid components: DOTAP: 40% SM-102: 30%; DSPC: 6%; (Cholesterol): 22% DMG-PEG2000: 1.5%; DSPE-PEG2000-Mal: 0.5%; In this embodiment, the sources of each raw material are as follows: CD31 antibody: Abcam (ab182981); SM-102: Aiweituo (2089251-47-6); DOTAP: Ruixi Biotechnology (LP-R4-117); DMG-PEG2000: Ruixi Biotechnology (R-PEG-0021); DSPE-PEG2000-Mal: Ruixi Biotechnology (R-0039-2k); mRNA encoding PIEZO2 protein: Yaohai Biotechnology (lot: 20250506); (Cholesterol): Ruixi Biotechnology (RH-100001); DSPC: Ruixi Biotechnology (LP-R4-076); The preparation steps of the delivery system in this embodiment include: S1, dissolve ionizable cationic lipids, cationic lipids, and PEGylated lipids in an organic solvent; Specifically: The preparation was carried out using a microfluidic method, with 3.69 mg DOTAP, 2.81 mg SM-102, 0.63 mg DSPC, 1.12 mg Cholesterol, 0.50 mg DMG-PEG2000, 0.19 mg DSPE-PEG2000-Mal added to the reaction vessel, and the total amount of ethanol used was 660 μL.

[0026] S2 was hydrated with acidic buffer (sodium acetate (50 mM), pH 4.0) and then homogenized by an extruder to form a blank LNP; Specifically: The total amount of diluted mRNA used was 1980ul (including 480ul mRNA and 1500ul sodium acetate solution (pH=4)).

[0027] S3, encapsulates the mRNA encoding PIEZO2 (PIEZO2 mRNA for short) inside the LNP; Specifically: The provided PIEZO2 mRNA was dissolved in a 10 mM sodium acetate solution at pH 3-4. Microfluidic mixing and encapsulation were performed at a mRNA:lipid (volume ratio) of 3:1, with a total flow rate of 12 mL / min. After preparation, buffer replacement, concentration, and 0.22 μm sterile filtration were performed.

[0028] The encapsulation efficiency and concentration of the LNP-PIEZO2 mRNA product (LNP encapsulated with PIEZO2 mRNA) were then detected using RiboGreen; particle size, PDI, and zeta potential were detected using the DLS method.

[0029] S4. After purifying the LNP containing PIEZO2 mRNA, it was mixed with CD31 antibody treated with a reducing agent (exposing thiol groups) under mild conditions to obtain the antibody-conjugated LNP-PIEZO2 mRNA complex. Specifically, the steps for preparing antibody-conjugated LNP-PIEZO2 mRNA include: S41, 144 μg (0.96 nmol) of full-length anti-mouse CD31 monoclonal antibody was reacted with 11.52 nmol (2.66 μg) of N-succinimide S-acetyl mercaptoacetate at room temperature for 1 h. After the reaction, the buffer was replaced using an ultrafiltration tube (Millipore UFC505024) to remove unreacted N-succinimide S-acetyl mercaptoacetate. Then, the resulting antibody (10 μL) was mixed with 0.5 M hydroxylamine hydrochloride (100 μL) and reacted at room temperature for 2.5 h to deprotect and expose the thiol groups for coupling. After the reaction, the buffer was replaced to obtain the antibody with exposed thiol groups for later use.

[0030] In step S42, the antibody with exposed thiol groups was reacted with 180 μg of LNP-PIEZO2 mRNA obtained in step S3 at room temperature for 3 h. After the reaction was completed, ultrafiltration was performed using an ultrafiltration tube (Sartorius VS2052) for purification. After purification, 0.22 μm sterile filtration was performed to obtain the antibody-conjugated LNP-PIEZO2 mRNA complex (i.e., the delivery system).

[0031] The encapsulation efficiency and concentration of the antibody-conjugated LNP-PIEZO2 mRNA complex were then determined using RiboGreen assay; particle size, PDI, and zeta potential were determined using the DLS method.

[0032] Example 2 In this embodiment, the LNP prepared in Example 1 was characterized; 1) Particle size distribution of LNP; LNP particle size distribution as shown in... Figure 2 As shown, the particle size distribution is between 80-100 nm, and PD=0.15, indicating good uniformity of particle size distribution.

[0033] 2) Characterization of the Zeta potential of LNPs: The characterization results are as follows Figure 2 As shown, the Zeta potential result is 8.535mV, which meets the qualified standard.

[0034] 3) mRNA encapsulation efficiency characterization: The encapsulation efficiency characterization process is as follows: LNP-mRNA encapsulation rate reflects the effective encapsulation degree of mRNA by lipid nanoparticles and is a key indicator determining delivery efficiency. It is usually detected using the Ribogreen fluorescence method. Encapsulation rate % = (RNA concentration after demulsification - RNA concentration before demulsification) / RNA concentration after demulsification × 100%. Characterization results are as follows... Figure 2 As shown, the encapsulation rate reaches 96.8% (≥90), which meets the standard.

[0035] Characterization results as follows Figure 2 As shown in the figure, it can be seen that Piezo2-mRNA-LNP meets the qualified standards in terms of concentration, encapsulation efficiency, particle size, polydispersity index (PDI), and zeta potential.

[0036] 4) Characterization of antibody conjugation efficiency: The process of antibody conjugation efficiency includes: Antibody conjugation efficiency is used to quantitatively assess the actual conjugation ratio between the antibody and the LNP vector, reflecting the targeting modification efficiency. It is usually detected using HPLC, and the antibody conjugation efficiency (%) is calculated as: (Total antibody amount − Free antibody amount) / Total antibody amount × 100%.

[0037] The characterization results are as follows: Figure 3 As shown, CD31-Piezo2-mRNA-LNP meets the qualified standards in terms of concentration, encapsulation efficiency, particle size, polydispersity index (PDI), and zeta potential.

[0038] Example 3 In this embodiment, in vitro cell targeting and transfection experiments were conducted on the delivery system prepared in Example 1; To evaluate the in vitro transfection capability of the constructed CD31-targeting lipid nanoparticle (CD31-LNP) delivery system on lung endothelial cells, PMVEC was used in the experiment. PMVEC was seeded in 12-well cell culture plates at a seeding density of 5 × 10⁶ cells / well. 5 Cells were cultured overnight at 37°C in a 5% CO2 incubator to allow them to adhere and reach approximately 60%–70% confluence. Then, a CD31-LNP delivery system carrying PIEZO2-mRNA was added to each well to a final concentration of 100–200 ng mRNA / well. After incubation at 37°C for 4 h, the culture medium was discarded and replaced with fresh complete culture medium for further culturing. Experimental setups included a Ctrl group, an LPS group, and an LPS+CD31-LNP group to compare the transfection efficiency of the delivery system. 24 h after transfection, intracellular PIEZO2 expression was observed using a fluorescence microscope.

[0039] The results are as follows Figure 4As shown, where, Figure 4 In the figure, A represents the expression of PIEZO2 in PMVEC treated with LPS via immunofluorescence detection of CD31-LNP delivery. The results showed that the decrease in PIEZO2 expression caused by LPS could be reversed by PIEZO2-mRNA delivery via CD31-LNP. Figure 4 In the figure, B indicates that the expression of PIEZO2 in PMVEC was detected using Western blot, and the results are consistent with... Figure 4 As verified in Example A, CD31-LNP delivery of PIEZO2-mRNA can reverse the LPS-induced decrease in PIEZO2 expression; it can be seen that the CD31-LNP treatment group exhibits a stable PIEZO2 expression level in mouse lung microvascular endothelial cells. This demonstrates that the CD31-targeted LNP delivery system constructed in Example 1 of this invention can promote the stable delivery and expression of nucleic acids in PMVECs.

[0040] Example 4 In this embodiment, an in vivo experiment was conducted on the delivery system prepared in Example 1; To further validate the in vivo lung endothelial targeting capability of the constructed CD31-targeting lipid nanoparticle (CD31-LNP) delivery system, in vivo distribution experiments were conducted using C57BL / 6 mice. Six- to eight-week-old male C57BL / 6 mice were randomly divided into a standard LNP group and a CD31-LNP group. LNPs pre-loaded with fluorescently labeled nucleic acids (such as Cy5-labeled mRNA) were administered to mice via tail vein injection at a dose of 0.5–1 mg / kg. Mice were sacrificed 24 hours after injection, and major organs, including the lungs, liver, spleen, kidneys, and heart, were isolated. Whole-organ fluorescence imaging was performed using a small animal in vivo fluorescence imaging system to evaluate the distribution of different delivery systems in each organ. Subsequently, frozen sections of lung tissue were prepared and immunofluorescence stained with the vascular endothelial marker CD31. Cellular localization of the delivery system in lung tissue was observed using confocal microscopy. Experimental results are as follows Figure 5 As shown; imaging results indicate that, compared with the ordinary LNP group, the CD31-LNP group exhibited stronger fluorescence signals in lung tissue, while fluorescence signals in other organs were significantly reduced; as shown Figure 6 As shown, confocal microscopy observation of lung tissue sections revealed that the CD31-Piezo2-LNP fluorescence signal highly overlapped with the stained area of ​​the vascular endothelial marker CD31, indicating that the delivery system is mainly located in the pulmonary vascular endothelial region.

[0041] This demonstrates that the CD31-targeted LNP delivery system constructed in this invention can achieve effective targeted delivery to pulmonary vascular endothelial cells in vivo, while reducing non-specific accumulation in the liver.

[0042] Example 5 In this embodiment, a pharmacodynamic experiment for treating lung injury was conducted on the delivery system prepared in Example 1. First, an acute lung injury model was established in mice (using airway infusion of LPS 10 mg / kg to establish the lung injury model). Different formulations were administered via tail vein to construct the following experimental groups: Control group: PBS; Group 1: Non-targeting LNP-PIEZO2 mRNA (unconjugated antibody CD31); Group 2: Targeting LNP-PIEZO2 mRNA (Example 1); To evaluate the therapeutic effect of the constructed CD31-targeted LNP delivery system on acute lung injury in vivo, an acute lung injury model in mice was established by lipopolysaccharide (LPS) airway instillation. Male C57BL / 6 mice aged 6–8 weeks were randomly divided into three groups: a PBS control group, a non-targeted LNP-PIEZO2 mRNA group, and a CD31-LNP-PIEZO2 mRNA-targeted group. The non-targeted LNP-PIEZO2 mRNA group consisted of LNPs encapsulated with PIEZO2 mRNA without antibody conjugation. Mice were anesthetized by intraperitoneal injection of sodium pentobarbital (50 mg / kg) and then administered LPS (10 mg / kg, dissolved in 50 μL PBS) via airway instillation to establish the acute lung injury model. Four hours after LPS treatment, appropriate treatments were administered via tail vein injection: the PBS group received an equal volume of PBS, while the other groups received the corresponding LNP formulations. Mice were sacrificed 24 hours after administration, and lung tissue was collected for relevant parameter detection. First, lung tissue was fixed, embedded in paraffin, and stained with hematoxylin and eosin (HE). The structural changes in lung tissue were observed under an optical microscope, and the pathological score of lung injury was determined based on indicators such as alveolar structural destruction, inflammatory cell infiltration, and interstitial edema. Tight junctions between lung endothelial cells were observed using transmission electron microscopy. The expression level of PIEZO2 protein in lung tissue was detected by Western blot.

[0043] Post-treatment assessment included lung tissue pathological damage scoring, transmission electron microscopy observation of tight junctions in lung endothelium, and PIEZO2 protein expression levels in lung tissue; experimental results are as follows: Figure 7 As shown, this CD31-targeted LNP delivery system can effectively deliver PIEZO2 mRNA to lung tissue and improve LPS-induced acute lung injury; the specific analysis is as follows: After administering the same dose of the PIEZO2 mRNA delivery system, there was a significant difference in therapeutic efficacy between the non-targeted LNP-PIEZO2 mRNA group (Group 1) and the targeted CD31-LNP-PIEZO2 mRNA group (Group 2). Compared with Group 1, the pathological damage in the lung tissue of mice in Group 2 was significantly reduced. HE staining showed a significant reduction in alveolar structural destruction, inflammatory cell infiltration, and interstitial edema. Transmission electron microscopy revealed that, compared with Group 1, the endothelial tight junctions in Group 2 were intact, and the mitochondrial structure was complete. Furthermore, Western blot analysis showed that the expression level of PIEZO2 protein in the lung tissue of Group 2 was significantly higher than that in Group 1, indicating that the targeted delivery system can more effectively promote the expression of PIEZO2 mRNA in lung tissue. The above results indicate that conjugating CD31 antibody to the surface of LNP can significantly improve the targeting ability of the delivery system to pulmonary vascular endothelial cells, thereby enhancing the delivery efficiency and expression level of PIEZO2 mRNA in lung tissue and further improving its therapeutic effect on LPS-induced acute lung injury. This suggests that CD31 targeting modification plays an important role in the delivery system of this invention.

[0044] In the description of this specification, references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0045] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the claimed invention.

Claims

1. A PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP, characterized in that, include: LNP, mRNA encoding PIEZO2 protein encapsulated within LNP, and CD31 antibody conjugated to the surface of LNP.

2. The PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP according to claim 1, characterized in that, The LNP comprises the following molar percentage of lipid components: Ionizable cationic lipids: 30%; Cationic lipids: 40%; PEGylated lipids: 2%; DSPC: 6%; Cholesterol: 22%.

3. The PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP according to claim 2, characterized in that, The ionizable cationic lipid is SM-102.

4. The PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP according to claim 2, characterized in that, The cationic lipid is DOTAP.

5. The PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP according to claim 2, characterized in that, The PEGylated lipids include: neutral PEGylated lipids and DSPE-PEG2000-Mal; the neutral PEGylated lipids are DMG-PEG2000 or DSPE-PEG.

6. The PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP according to claim 5, characterized in that, The molar percentage of DSPE-PEG2000-Mal is 0.5% of LNP.

7. The PIEZO2 mRNA lung endothelial cell targeted delivery system based on antibody-conjugated LNP according to claim 3, characterized in that, The CD31 antibody is covalently coupled to the maleimide group at the end of DSPE-PEG-Mal via a thiol group.

8. The method for preparing the antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system according to any one of claims 1-7, characterized in that, Includes the following steps: S1, dissolve ionizable cationic lipids, cationic lipids and PEGylated lipids in an organic solvent, remove the solvent, and form a lipid film; S2, the lipid membrane was hydrated with acidic buffer solution and then homogenized to form a blank LNP; S3, encapsulate the mRNA encoding PIEZO2 inside the LNP; S4. After purifying the LNP containing PIEZO2 mRNA, it is mixed with CD31 antibody treated with a reducing agent to obtain the antibody-conjugated LNP-PIEZO2 mRNA complex, i.e., the delivery system.

9. The use of the antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system according to any one of claims 1-7 in the preparation of drugs for treating or preventing lung injury.

10. A medicament for treating or preventing lung injury, characterized in that, include: The antibody-conjugated LNP-based PIEZO2 mRNA lung endothelial cell targeted delivery system as described in claims 1-7.

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