Application of PDLIM2 gene overexpression viral vector in the preparation of drugs for treating podocyte disease

Abstract

This invention discloses the application of a PDLIM2 gene overexpression viral vector in the preparation of drugs for treating podocyte disease, wherein the nucleotide sequence of the PDLIM2 gene is shown in SEQ ID NO:1. This invention is the first to discover and verify the core role of the PDLIM2 gene in podocyte protection, confirming its significant downregulation in DKD and FSGS disease models, and demonstrating through functional experiments that PDLIM2 overexpression can effectively stabilize the podocyte cytoskeleton, inhibit apoptosis, and reduce proteinuria, thus establishing it as a key target gene for the treatment of podocyte disease. This invention is the first to intervene at the gene level in the core link of podocyte damage—cytoskeleton stability. By specifically upregulating the expression of PDLIM2 in podocytes, it directly enhances their intrinsic cytoskeleton support and anti-damage ability, achieving a fundamental shift in treatment strategy from "symptomatic support" to "causal repair."

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to the application of a PDLIM2 gene overexpression viral vector in the preparation of a drug for treating podocyte disease. Background Technology

[0002] Podocyte injury is a core driving event in the development of many progressive glomerular diseases. Diabetic Kidney Disease (DKD) and primary or secondary focal segmental glomerulosclerosis (FSGS) are two key causes of end-stage renal disease in clinical practice, seriously threatening patients' lives and health and imposing a heavy medical burden. In the pathological process of DKD, multiple factors such as the hyperglycemic microenvironment, systemic metabolic disorders, and abnormal renal hemodynamics work synergistically to induce a series of pathological changes in podocytes, including metabolic imbalance, inflammation activation, abnormal hypertrophy, and apoptosis, ultimately leading to progressive loss of podocytes and disruption of the glomerular filtration barrier integrity. FSGS, on the other hand, is characterized by severe podocyte degeneration, detachment, and segmental glomerular scarring. Its pathogenesis involves multiple levels, including immune disorders, genetic defects, and circulating pathogenic factors, and different pathogenic pathways ultimately point to the common pathological endpoint of podocyte skeletal structure destruction and loss of cell adhesion function.

[0003] Currently, clinical treatment for DKD and FSGS still faces significant challenges, as existing therapies struggle to precisely target and repair damaged podocytes. For DKD, clinical treatment primarily relies on systemic glycemic regulation, renin-angiotensin system blockers, and the recently used SGLT2 inhibitors. While these therapies can slow disease progression to some extent, they lack the ability to specifically reverse or repair damaged podocytes, and their efficacy is particularly limited for patients in advanced stages. For FSGS, first-line treatment involves high-dose glucocorticoids combined with calcineurin inhibitors (such as cyclosporine and tacrolimus). However, 30%-50% of patients exhibit hormone resistance or dependence, and the relapse rate remains high. Furthermore, long-term use of these drugs can lead to serious infections, nephrotoxicity, and other adverse reactions, further exacerbating the patient's condition. In summary, both DKD and FSGS lack specific treatments that can directly target the intrinsic damage mechanisms of podocytes and effectively promote podocyte repair and survival. Developing innovative therapies that precisely target podocytes has become a critical clinical need.

[0004] As research progresses, the stability of the actin cytoskeleton within podocytes is considered a cornerstone for maintaining their normal function and resisting damage. In the pathological processes of DKD and FSGS, multiple damage signals lead to the degradation of podocyte cytoskeleton proteins (such as synaptopodin), cytoskeleton depolymerization, and stress fiber formation, ultimately resulting in foot process loss and proteinuria. Therefore, identifying key regulatory factors capable of endogenously stabilizing the podocyte cytoskeleton is a strategic direction for developing breakthrough therapies. Summary of the Invention

[0005] The main objective of this invention is to propose the application of a PDLIM2 gene overexpression viral vector in the preparation of drugs for treating podocyte diseases. This aims to address the problems of existing treatments for DKD and FSGS, such as their inability to directly repair podocytes, significant side effects, and poor efficacy in refractory cases. The invention provides a precision gene therapy based on a PDLIM2 overexpression virus that can enhance the resistance and stability of podocytes from within the cells, offering a novel and validated technical solution for developing innovative drugs for treating DKD and FSGS.

[0006] To achieve the above objectives, this invention proposes the application of a viral vector overexpressing the PDLIM2 gene in the preparation of a drug for treating podocyte disease, wherein the nucleotide sequence of the PDLIM2 gene is as shown in SEQ ID NO:1.

[0007] Preferably, the podocyte disease includes diabetic nephropathy and focal segmental glomerulosclerosis.

[0008] Preferably, the PDLIM2 gene overexpression viral vector uses pLV3-CMV-MCS-3×FLAG-CopGFP-Puro as the vector backbone.

[0009] Preferably, the drug is administered via local renal injection, with an injection dose of 5 × 10⁻⁶. 6 U / kidney.

[0010] Preferably, the drug is used to reduce the urinary albumin-to-creatinine ratio in patients or animal models of podocyte disease, thereby alleviating glomerular filtration barrier dysfunction.

[0011] Preferably, the drug is used to improve the fusion and disappearance of podocyte foot processes and protect the ultrastructure of podocytes.

[0012] Preferably, the drug is used to restore the expression of podocyte markers podocin and WT1.

[0013] Preferably, the drug is used to alleviate symptoms of glomerular sclerosis, kidney enlargement, and increased kidney weight ratio in patients or animal models of podocyte disease.

[0014] The present invention also proposes a pharmaceutical composition comprising lentiviral particles overexpressing the PDLIM2 gene, wherein the lentiviral particles are obtained by co-transfecting packaging cells with the PDLIM2 gene overexpressing viral vector, packaging plasmid, and envelope plasmid as described above.

[0015] Preferably, the packaging plasmid is psPAX2, the envelope plasmid is pMD2.G, and the packaging cells are HEK293T cells.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0017] (1) The technical solution provided by this invention first discovered and verified the core role of the PDLIM2 gene in podocyte protection, confirmed that its expression was significantly downregulated in DKD and FSGS disease models, and demonstrated through functional experiments that PDLIM2 overexpression can effectively stabilize the podocyte cytoskeleton, inhibit apoptosis, and reduce proteinuria, thus establishing it as a key target gene for the treatment of podocyte disease. Then, through molecular cloning technology, the full-length coding sequence of the murine PDLIM2 gene was cloned into a lentiviral transfer plasmid vector to construct the pLV-PDLIM2 recombinant overexpression vector. The PDLIM2 gene overexpression viral vector provides an innovative solution for the treatment of podocyte disease that is precisely targeted and directly addresses the core pathology, significantly improving the effectiveness and durability of treatment. This invention is the first to intervene at the gene level in the core link of podocyte damage—cytoskeleton stability. By specifically upregulating the expression of PDLIM2 in podocytes, the intrinsic cytoskeleton support and anti-damage ability are directly enhanced, realizing a fundamental shift in treatment strategy from "symptomatic support" to "causal repair." This breaks through the bottleneck of current therapies with indirect target action and limited efficacy, laying the foundation for precision treatment of podocyte diseases.

[0018] (2) Compared with traditional symptomatic treatment drugs, the lentiviral vector used in this invention stably integrates the PDLIM2 gene into the host cell genome, achieving long-term sustained expression of the target gene. A single dose can achieve long-term therapeutic effects, effectively curbing the progression and recurrence of chronic podocyte disease. It has breakthrough therapeutic value, especially for refractory cases such as hormone-resistant FSGS and advanced DKD, filling a gap in clinical treatment. The lentiviral vector used in this invention delivers the drug through multi-site injection into the renal cortex. Combined with the natural tropism of lentiviruses to kidney tissue and their ability to infect non-dividing cells, it can precisely target podocytes to exert its effect, minimizing the impact of the drug on other tissue systems throughout the body and avoiding the toxic side effects and drug resistance risks associated with traditional systemic administration (such as immunosuppressants). The lentiviral vector used in this invention delivers the PDLIM2 gene to glomerular podocytes specifically, which can effectively reverse the pathological state of downregulated PDLIM2 expression in podocyte diseases (such as DKD and FSGS). By stabilizing the podocyte cytoskeleton and inhibiting podocyte apoptosis, it can repair the damaged glomerular filtration barrier from the root, significantly reduce the urinary albumin-to-creatinine ratio, and alleviate key pathological damage such as glomerular sclerosis and podocyte fusion. 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, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 This is a staining diagram showing the expression of PDLIM2 in the kidney tissues of patients with DKD (A) and FSGS (B) according to the present invention.

[0021] Figure 2 A: Flowchart for constructing the DKD mouse model; B: Quantitative statistical graph of PDLIM2 gene expression in the renal cortex of DKD mice; C: Graph of PDLIM2 expression level in the renal tissue of DKD mice detected by Western blotting; D: Quantitative statistical graph of PDLIM2 protein expression level in the renal tissue of DKD mice.

[0022] Figure 3A: Flowchart for constructing the FSGS mouse model; B: Quantitative statistical graph of PDLIM2 gene expression in the renal cortex of FSGS mice; C: Graph of PDLIM2 expression level detected by Western blotting in the renal tissue of FSGS mice; D: Quantitative statistical graph of PDLIM2 protein expression level in the renal tissue of FSGS mice.

[0023] Figure 4 This is a construction map of the PDLIM2 overexpression recombinant lentiviral vector of the present invention.

[0024] Figure 5 The diagram shows the efficacy evaluation of PDLIM2 overexpressing lentivirus in in vivo treatment of DKD; A: Schematic diagram of the experimental procedure for the DKD model; B: Immunofluorescence staining diagram for detecting PDLIM2 expression in the kidney; C: RT-qPCR quantitative statistical diagram for detecting PDLIM2 expression in the kidney; D: Statistical diagram of urine albumin-creatinine ratio in mice of each treatment group; E: Kidney weight / body weight ratio in mice of each treatment group; F: Detection diagram of podocyte markers (Podocin, WT1) in mice of each treatment group; G: Transmission electron micrograph of podocyte ultrastructure in mice of each treatment group.

[0025] Figure 6 A: Schematic diagram of the efficacy evaluation of PDLIM2 overexpressing lentivirus in in vivo treatment of FSGS; B: Statistical diagram of urinary albumin-creatinine ratio in mice of each treatment group; C: PAS staining diagram of glomerular pathological changes in mice of each treatment group; D: Detection diagram of podocyte markers (Podocin, Nephrin, WT1) in mice of each treatment group; E: Transmission electron micrograph of podocyte ultrastructure in mice of each treatment group.

[0026] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially. Furthermore, the technical solutions of the various embodiments can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention. 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.

[0028] The technical solution of the present invention will be further described in detail below with reference to specific embodiments and accompanying drawings. It should be understood that the following embodiments are only used to explain the present invention and are not intended to limit the present invention.

[0029] Example 1: Detection of PDLIM2 expression in renal tissues of patients with DKD and FSGS

[0030] To clarify the correlation between PDLIM2 and human podocyte disease, immunohistochemistry was used to detect its presence in paraffin sections of kidneys from patients with DKD and FSGS, with adjacent normal kidney tissue from patients undergoing radical nephrectomy serving as a control. Sections were dewaxed with xylene, hydrated with graded ethanol, and then subjected to 3% hydrogen peroxide solution to eliminate endogenous peroxidase activity. Microwave retrieval was performed using citrate antigen retrieval buffer to expose the antigen. The sections were then blocked with 5% donkey serum at room temperature for 1 hour, followed by incubation at 4°C overnight with anti-PDLIM2 antibody (1:1000 dilution). The next day, after washing with PBS, biotinylated secondary antibody was added and incubated at room temperature for 1 hour. After washing again, DAB reagent was used for staining, and the nuclei were counterstained with hematoxylin. Finally, the sections were mounted with aqueous mounting medium. Under an optical microscope, the positive signal appeared as brownish-red granules. Compared with normal kidney tissue, the positive staining signal of PDLIM2 in the glomeruli of patients with DKD and FSGS was significantly weakened. Figure 1 (A, B). This result is the first clinical-sample-level confirmation that PDLIM2 expression is downregulated in the kidneys of patients with podocyte disease, suggesting that its expression deficiency may be associated with disease development and progression.

[0031] Example 2: Detection of PDLIM2 expression in kidney tissue of DKD model mice

[0032] To validate the correlation between PDLIM2 and DKD at the animal model level, this study further examined its expression in the kidneys of DKD mice. Seven-week-old SPF-grade male C57BL / 6J mice were used for modeling. Mice were randomly assigned to a control group (Sham group) and a model group (DKD group). The DKD group received a high-fat diet for 4 weeks, while the Sham group received a standard control diet. Unilateral nephrectomy was then performed; the right kidney was removed in the DKD group, and a sham operation was performed in the Sham group. One week post-surgery, mice were fasted for 8 hours and then administered streptozotocin (STZ, 50 mg / kg, injected protected from light) intraperitoneally, three times a week. The Sham group received an equal dose of the STZ solvent—citrate buffer—intraperitoneally. For the next 12 weeks, both the DKD and Sham groups continued to receive either a high-fat diet (HFD) or a standard control diet. Random blood glucose levels were measured within two weeks of STZ administration; a successful diabetes model was defined as a random blood glucose level above 16.7 mmol / L. Kidney tissue was collected after successfully constructing the DKD model. Figure 2 A). RNA was extracted from mouse renal cortex using the Trizol-chloroform-isopropanol method, and the expression of the PDLIM2 gene was detected by real-time quantitative PCR after reverse transcription. The results showed that the expression of PDLIM2 gene was downregulated in the renal cortex of DKD mice. Figure 2 B). Further verification was performed using Western blotting, and total protein was extracted from the renal cortex for detection. The results showed that the expression level of PDLIM2 protein in the renal tissue of DKD mice was significantly downregulated (B). Figure 2 C, D).

[0033] Example 3: Detection of PDLIM2 expression in kidney tissue of FSGS model mice

[0034] To validate the correlation between PDLIM2 and FSGS in an animal model, this study further examined its expression in the kidneys of FSGS mice. Seven-week-old SPF-grade male C57BL / 6J mice were used for modeling. Mice were randomly divided into a control group (Ctrl group) and a model group (FSGS group). Doxorubicin (15 mg / mL) was prepared based on mouse weight and administered via tail vein, while the Ctrl group received an equal volume of saline. After the single injection, the mice were kept in an SPF-grade environment for 5 weeks, after which tissue samples were collected. Figure 3 A). RNA was extracted from mouse renal cortex using the Trizol-chloroform-isopropanol method, and the expression of the PDLIM2 gene was detected by real-time quantitative PCR after reverse transcription. The results showed that the expression of PDLIM2 gene was downregulated in the renal cortex of FSGS mice. Figure 3B). Further verification was performed using Western blotting, and total protein was extracted from the renal cortex for detection. The results showed that the expression level of PDLIM2 protein in the renal tissue of FSGS mice was significantly downregulated (B). Figure 3 C, D).

[0035] Example 4: Construction of PDLIM2 overexpression lentivirus

[0036] 1. Obtaining the nucleotide sequence of the PDLIM2 gene

[0037] The nucleotide sequence (SEQ ID NO:1) of the mouse PDLIM2 gene was obtained by searching the NCBI GenBank database (www.ncbi.nlm.nih.gov / gene) for transcript NM_001253736.1:

[0038] SEQ ID NO:1:

[0039]

[0040] 2. Obtaining cDNA

[0041] Total RNA was extracted from immortalized mouse podocytes (MPCs) using an RNA extraction kit. The RNA was then reverse transcribed into a cDNA library using the following reaction system: PrimeScriptRT Buffer (8 μL), RT primer Mix (8 μL), PrimeScriptRT Enzyme Mix I (2 μL), and RNase-free dH2O (2 μL) at 37°C for 15 min followed by 85°C for 5 s.

[0042] 3. PCR amplification of the target fragment

[0043] Based on the obtained PDLIM2 gene nucleotide sequence, a pair of specific primers targeting the PDLIM2 gene were designed and synthesized. The forward primer sequence is 5'-CAGCCCAGTTGCCCTTTCTA-3' (SEQ ID NO:2); the reverse primer sequence is 5'-GTTCGAGATGTTGACGCTGC-3' (SEQ ID NO:3). PCR amplification system: 25 μL 2X PCR Buffer, 2 μL dNTPs mix (10 mM each), 2 μL forward primer (10 μM), 2 μL reverse primer (10 μM), 1 μL cDNA template (200 ng / μL), 18 μL ddH2O, 1 μL phanta Super-Fidelity DNA Polymerase. Set the PCR reaction conditions for amplification: pre-denaturation at 95℃ for 5 min, denaturation at 95℃ for 30 s, annealing at 60℃ for 30 s, extension at 72℃ for 30-60 s / kb (27-35 cycles), and final extension at 72℃ for 10 min to obtain the PCR product.

[0044] 4. Construction of PDLIM2 overexpression lentivirus

[0045] The pLV3-CMV-MCS-3×FLAG-CopGFP-Puro vector was used as the backbone. The PCR product and the pLV3-CMV-MCS-3×FLAG-CopGFP-Puro vector were digested with EcoRI and BamHI, respectively. After digestion, agarose gel electrophoresis was performed to recover the target fragment. Then, the vector and the target fragment were ligated using T4 DNA ligase to obtain the PDLIM2 overexpression recombinant lentiviral vector. The constructed vector map is shown below. Figure 4 As shown.

[0046] 5. Plasmid amplification, preliminary screening, and verification of positive clones.

[0047] The overexpression lentiviral vector was transformed into competent bacteria, then streaked onto LB agar plates containing puromycin (100 μg / mL). Single clones were picked after culture, plasmids were extracted in small quantities, identified by double enzyme digestion, and positive clones were sent for sequencing verification to obtain the validated PDLIM2 overexpression lentiviral vector. The original pLV3-CMV-MCS-3×FLAG-CopGFP-Puro vector (without the PDLIM2 gene inserted) was used as the empty control vector LV-Vector.

[0048] 6. Virus Packaging

[0049] Lentiviral packaging was performed using the HEK293T cell line. Following the instructions of the Lipofiter™ transfection reagent (Shanghai Hanheng Biotechnology Co., Ltd.), 10 μg of the PDLIM2 overexpression recombinant lentiviral vector or 10 μg of the pLV3-CMV-MCS-3×FLAG-CopGFP-Puro empty vector was co-transfected into HEK293T cells with 10 μg of the pSPAX2 packaging plasmid and 5 μg of the pMD2G envelope plasmid, respectively. 48 h post-transfection, cell culture medium was collected, centrifuged at 2000×g for 10 min at 4°C to remove cell debris, and the supernatant was then ultracentrifuged at 82700×g for 120 min at 4°C. The viral pellet was collected and resuspended in complete culture medium to obtain either a concentrated PDLIM2 overexpression lentiviral solution or a concentrated control lentiviral solution, which was stored at -80°C for later use.

[0050] 7. Titer determination

[0051] The viral titers (TU / mL) of PDLIM2 overexpressing lentivirus (LV-PDLIM2) and empty vector control lentivirus (LV-Vector) were determined based on the expression levels of the fluorescent protein in HEK293T cells. The specific procedures are as follows:

[0052] The virus was serially diluted. The dilution method was as follows: Prepare eight 1.5 mL sterile EP tubes for each virus. Add 297 μL of complete culture medium to each tube. Add 33 μL of the original virus solution to the first tube, mix well, and then add 30 μL to the second tube and mix well. Repeat this process to create eight dilutions (10~10E-6). HEK293T cells pre-seeded in 96-well plates (1 × 10⁶ cells per well) were then infected. 4(cells). Discard the original culture medium in the 96-well plate. Repeat each dilution in triplicate, adding 100 μL of diluted virus solution to each well. Label the wells. After 72 h, perform fluorescence counting on HEK293T cells using a fluorescence microscope. Record the number of fluorescent cells in the last two wells where fluorescence is visible. Calculate the sum of the total number in the three replicate wells and calculate the average, denoted as A (average number of fluorescent cells in the second-to-last visible well) and B (average number of fluorescent cells in the last visible well). The formula for calculating lentivirus titer is shown below:

[0053] Viral titer (TU / mL) = (A + B × 10) × 1000 / 2 / Viral load in well A (μL)

[0054] The final measured LV-PDLIM2 viral titer was 8.59 × 10⁻⁶. 8 TU / mL, LV-Vector viral titer is 3.62×10⁻⁶. 9 TU / mL.

[0055] Example 5: Evaluation of the efficacy of PDLIM2 overexpression lentivirus in in vivo treatment of DKD

[0056] To evaluate the therapeutic effect of PDLIM2 overexpressing lentivirus in a complete physiological environment, this study conducted an in vivo treatment experiment in a DKD animal model to systematically evaluate its ameliorative effect on key disease phenotypes. First, a DKD model was established using 7-week-old SPF-grade male C57BL / 6J mice according to the DKD model construction method described in Example 2. Four weeks prior to sampling, mice were randomly divided into four groups: ① sham + LV-Vector group; ② sham + LV-PDLIM2 group; ③ DKD + LV-Vector group; ④ DKD + LV-PDLIM2 group. All mice were injected with the virus according to their groups. Specifically, LV-PDLIM2 or LV-Vector was injected into the upper and lower poles of the left kidney, with 5 × 10⁶ liters injected into each kidney. 6 U. After modeling, blood, urine, and kidneys of mice were collected. Figure 5 A). Immunofluorescence staining and RT-qPCR results showed that PDLIM2 was successfully overexpressed in the kidney. Figure 5 B, C). Compared with DKD mice, mice treated with PDLIM2 lentivirus showed a significantly lower urinary albumin-to-creatinine ratio (PGR). Figure 5 (D) indicates that it can effectively alleviate glomerular filtration barrier dysfunction. In addition, PDLIM2 lentiviral treatment can alleviate kidney enlargement and increased kidney weight ratio in DKD mice (D). Figure 5 E). Immunofluorescence and immunohistochemical staining results showed that PDLIM2 lentivirus restored the expression of podocyte markers podocin and WT1 (E). Figure 5F). Transmission electron microscopy results showed that PDLIM2 lentivirus protected podocytes in DKD mice, specifically by significantly improving the fusion and disappearance of podocyte foot processes compared to the control group, and by exhibiting a more orderly arrangement of foot process structures. Figure 5 G).

[0057] Example 6: Evaluation of the efficacy of PDLIM2 overexpression lentivirus in vivo treatment of FSGS

[0058] To verify the in vivo therapeutic effect of PDLIM2-overexpressing lentivirus on FSGS, this study constructed an doxorubicin-induced FSGS mouse model and conducted in vivo intervention. First, 7-week-old male C57BL / 6J mice were randomly divided into four groups: ① Ctrl+LV-Vector group; ② Ctrl+LV-PDLIM2 group; ③ FSGS+LV-Vector group; ④ FSGS+LV-PDLIM2 group. Before ADR modeling, LV-PDLIM2 or LV-Vector was injected into the upper and lower poles of the left kidney of each group, with 5 × 10⁻⁶ injections per kidney. 6 U. One week later, the FSGS model was established according to the FSGS model construction method in Example 3. Modeling was completed after 5 weeks, and mouse blood, urine, and kidney tissue were collected. Figure 6 A). The results showed that PDLIM2 overexpression lentivirus significantly reduced the urinary albumin-to-creatinine ratio in FSGS mice (A). Figure 6 B); at the same time, it also alleviated glomerular sclerosis ( Figure 6 C). Immunofluorescence staining and immunohistochemical staining results showed that PDLIM2 lentivirus restored the expression of podocyte markers podocin and WT1 (C). Figure 6 D). Transmission electron microscopy results showed that PDLIM2 overexpression lentivirus restored the foot process fusion and disappearance phenomenon in FSGS mice (D). Figure 6 E). The in vivo experimental results show that PDLIM2 overexpression lentiviral treatment can significantly improve proteinuria, pathological damage and podocyte ultrastructural disruption in FSGS model mice, thus confirming its therapeutic effect on FSGS at the in vivo level.

[0059] The above are merely preferred embodiments of the present invention and do not limit the patent scope of the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the patent protection scope of the present invention.

Claims

1. The application of a PDLIM2 gene overexpression viral vector in the preparation of drugs for treating podocyte disease, characterized in that, The nucleotide sequence of the PDLIM2 gene is as shown in SEQ ID NO:1, and the podocyte disease is diabetic nephropathy or focal segmental glomerulosclerosis.

2. The application according to claim 1, characterized in that, The PDLIM2 gene overexpression viral vector uses pLV3-CMV-MCS-3×FLAG-CopGFP-Puro as its vector backbone.

3. The application according to claim 1, characterized in that, The drug is administered via local renal injection, with an injection dose of 5 × 10⁻⁶. 6 U / kidney.

4. The application according to claim 1, characterized in that, The drug is used to reduce the urinary albumin-to-creatinine ratio in patients or animal models of podocyte disease, thereby alleviating glomerular filtration barrier dysfunction.

5. The application according to claim 1, characterized in that, The drug is used to improve the fusion and disappearance of foot processes in podocytes and to protect the ultrastructure of podocytes.

6. The application according to claim 1, characterized in that, The drug is used to restore the expression of podocyte markers podocin and WT1.

7. The application according to claim 1, characterized in that, The drug is used to alleviate symptoms of glomerular sclerosis, kidney enlargement, and increased kidney weight ratio in patients or animal models of podocyte disease.

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