Application of PDAP1 protein in preparation of medicine for regulating and controlling human lifespan-related diseases

Abstract

The invention relates to an application of PDAP1 protein in preparation of a medicine for regulating and controlling human life-related diseases, PDAP1 is used as a novel molecular target and is obviously related to various life-related diseases and characters; the biological function of the PDAP1 is comprehensively revealed by integrating genetic data of transcriptomics, proteomics and life-related traits; the PDAP1 action mechanism and the causal effect provide theoretical basis and technical support for research and development of drugs for life-related diseases in the future.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to an application of PDAP1 protein in the preparation of drugs for regulating diseases related to human life span. Background Art

[0002] Extending lifespan and healthy lifespan is a key goal in global public health. However, with the aging of the population, the incidence of age-related diseases such as cardiovascular disease, cancer, neurodegenerative diseases, and metabolic diseases has increased significantly, placing a huge burden on global healthcare systems.

[0003] Traditional research has focused on the pathological mechanisms of a single organ or disease. However, the essence of lifespan and healthspan involves the integrated effects of multiple systems and levels. Therefore, it is of great significance to identify the core molecular targets that influence lifespan and the aging process. In recent years, large-scale genome-wide association studies (GWAS) have revealed a variety of genetic loci associated with lifespan. However, the functions of these loci and how they regulate lifespan at the molecular level have not been fully explored.

[0004] Platelet-derived growth factor subunit A-associated protein 1 (PDAP1) is a ubiquitously expressed protein involved in multiple biological processes. However, the role of PDAP1 in lifespan and healthy aging has not been thoroughly investigated. Summary of the Invention

[0005] The technical problem addressed by this invention is the use of the PDAP1 protein in the preparation of drugs for regulating diseases related to human lifespan. By integrating multi-omics data (including gene expression, proteomics, and population lifespan data), this invention systematically reveals the significant impact of PDAP1 on lifespan and various related diseases, providing a scientific basis for the development of new drugs.

[0006] The present invention provides an application of a PDAP1 inhibitor in the preparation of anti-aging drugs.

[0007] The present invention provides an application of a PDAP1 inhibitor in the preparation of a drug for delaying cell aging.

[0008] The present invention provides an application of a PDAP1 inhibitor in the preparation of a drug for improving lifespan-related diseases, wherein the lifespan-related diseases to be improved are one or more of Alzheimer's disease, cognitive impairment, cardiovascular metabolic diseases, and the like.

[0009] The present invention provides an application of PDAP1 as a drug target in screening or preparing drugs for delaying cell aging.

[0010] The present invention provides an application of PDAP1 as a drug target in screening or preparing drugs for improving lifespan-related diseases.

[0011] The medicine also includes single-ingredient preparations or compound preparations.

[0012] The present invention provides an application of a biomarker in the preparation of a mortality risk and / or life expectancy diagnosis or auxiliary diagnosis product, wherein the biomarker is PDAP1.

[0013] The present invention provides use of a reagent for detecting a biomarker in preparing a product for diagnosing or assisting diagnosis of mortality risk and / or life expectancy, wherein the biomarker is PDAP1.

[0014] PDAP1 gene and protein levels were positively correlated with mortality risk and negatively correlated with life expectancy. For example, elevated plasma PDAP1 levels in the upper tertile or per unit PDAP1 level (normalized protein expression, NPX) significantly increased the risk of all-cause mortality. The mortality risk in the high-PDAP1 group was significantly higher than that in the low-PDAP1 group. The life expectancy of the high-PDAP1 group was approximately 0.84 years shorter than that of the low-PDAP1 group.

[0015] The present invention provides a new drug target, PDAP1 gene and its protein, for use in screening and preparing drugs for treating lifespan-related diseases.

[0016] Specifically, the purpose of the present invention is achieved through the following technical solutions:

[0017] The largest and most comprehensive proteome-wide genome-wide association study GWAS summary data (5 proteome research consortium GWAS summary data) and an independent large-sample validation protein genome data, GTEx V8 database and eQTLGenConsortium, covering gene expression levels in multiple tissues, were used as exposure factors;

[0018] Three lifespan-related traits were used to avoid single phenotypes: parental lifespan (continuous variable), the top 1% extreme longevity, and the top 10% extreme longevity (case-control traits), with data from large GWAS analyses.

[0019] Using Mendelian randomization and colocalization analysis, we found that PDAP1 gene and protein levels have significant causal effects on multiple lifespan-related traits (including lifespan, extreme longevity, etc.).

[0020] A variety of sensitivity analysis methods were used, including MR-Egger to assess potential horizontal pleiotropic bias, Cochran's Q test to detect heterogeneity, and Steiger directional test to verify whether the causal direction was reasonable.

[0021] At the same time, further mechanistic studies have shown that PDAP1 affects lifespan by regulating multiple physiological pathways (such as sex hormones, fat metabolism, and epigenetic aging acceleration).

[0022] In addition, the present invention verified the regulatory effect of PDAP1 on the aging process at the cellular level through the classic cell aging model, and verified the correlation between PDAP1 and all-cause mortality and life expectancy loss in a large-scale population cohort.

[0023] Beneficial effects

[0024] The present invention provides a complete set of new, efficient and reliable technical methods and processes for screening and clarifying intervention targets for lifespan-related diseases; it discovers and proves a protein gene target that has a significant causal effect on human lifespan and related chronic diseases, and has potential application value in drug development and clinical transformation.

[0025] Compared with the existing technology, the advantages of the present invention include: new target identification: PDAP1, as a new molecular target, is significantly associated with a variety of lifespan-related diseases and traits; multi-omics integration: by integrating transcriptomics, proteomics and genetic data of lifespan-related traits, the biological function of PDAP1 is fully revealed; clinical translation potential: the mechanism of action and causal effect of PDAP1 provide a theoretical basis and technical support for future drug development for lifespan-related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 :Causal association between PDAP1 protein and lifespan and association analysis between PDAP1 levels and mortality risk and lifespan in UK Biobank; (A) gene co-localization map (PDAP1 gene, ±500kb) shows the gene expression of PDAP1 in whole blood,

[0027] Plasma protein levels are linked to human lifespan outcomes, following the central dogma: eQTL / pQTL for PDAP1 (either in cis or trans) have significant effects on lifespan, with high colocalization evidence. (B) Effects of PDAP1 gene transcripts and plasma protein levels on mediating factors in the association with lifespan outcomes. Mediating factors that are beneficial to lifespan are shown in green (sex hormone-binding globulin SHBG and daily walking speed), and mediating factors that are detrimental to lifespan are shown in purple. (C) Effects of PDAP1 gene transcripts and plasma protein levels on the aging process (PhenoAge and GrimAge). (D) and (E) Association between a unit increase in plasma PDAP1 level and the risk of all-cause mortality. (F) Compared with the low-PDAP1 group, the loss of lifespan in the medium- and high-PDAP1 group is associated with medium- and high-PDAP1 levels;

[0028] Figure 2 : Changes in PDAP1 expression levels in cell senescence models; (A) Immunoblot images show PDAP1 protein levels in MRC-5 cells collected at different population doublings (PD). (B) Real-time quantitative reverse transcription-polymerase chain reaction analysis compares PDAP1 mRNA expression in MRC-5 cells at different PDs. (C) and (D) Comparison of PDAP1 mRNA expression in MRC-5 cells treated with UVA (C) or doxorubicin (D) and the control group. (E) Senescence-associated β-galactosidase results in MRC-5 cells at different concentrations of exogenous PDAP protein. Quantification of the percentage of senescence-associated β-galactosidase-positive cells. (F) and (G) Real-time quantitative reverse transcription-polymerase chain reaction analysis compares p16 (F) and p21 (G) mRNA expression in MRC-5 cells at different concentrations of exogenous PDAP protein. (H) EdU results in MRC-5 cells at different concentrations of exogenous PDAP protein. Quantification of the percentage of EdU-positive cells.

[0029] Figure 3 Results of functional validation of PDAP1 in cell senescence models: (A) Construction of a cell senescence model in which PDAP1 expression was silenced. (B) Effects of PDAP1 silencing on senescence-associated β-galactosidase at different population doublings (PDs). (C) Effects of PDAP1 silencing on p16 and p21 mRNA expression. (D) Effects of PDAP1 silencing on EdU. (E) Growth curve of MRC-5 cells at PD 60 in which PDAP1 was silenced by lentivirus. Cells were grown in 5% oxygen until senescence. DETAILED DESCRIPTION

[0030] Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiments are only used to illustrate the present invention and are not used in limiting the scope of the present invention.In addition, should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall equally within the scope limited by the appended claims of the application.

[0031] Example 1

[0032] Multi-omics analysis reveals a causal role for PDAP1

[0033] This example systematically reveals the causal effect of PDAP1 on lifespan and its related diseases by integrating multi-omics data (including genome-wide association study data of gene expression, plasma proteins and human lifespan-related traits) based on Mendelian randomization and co-localization analysis.

[0034] (1) Data source

[0035] This study used a variety of high-quality omics data, including:

[0036] Gene expression levels (eQTLs): Data comes from the GTEx V8 database and the eQTLGen Consortium, covering gene expression levels in multiple tissues.

[0037] Plasma protein levels (pQTLs): Data were integrated from plasma proteomic analyses of five independent studies, including INTERVAL, KORA, ARIC, and FHS.

[0038] Lifespan-related traits: including three lifespan-related traits: parental lifespan (continuous variable), top 1% extreme longevity and top 10% extreme longevity (case-control traits), and the data are from large-scale GWAS analysis.

[0039] (2) Analytical methods

[0040] Mendelian randomization analysis: Using genetic variation as instrumental variables to assess the causal effects of thousands of gene expressions and plasma protein levels, including PDAP1, on lifespan-related traits.

[0041] Colocalization analysis: We used colocalization methods to analyze whether tissue-specific gene transcriptional expression, plasma protein levels, and lifespan traits, such as PDAP1, share the same causal variant. We used the Moloc tool for multi-trait colocalization analysis, calculating the colocalization probability (Pabc > 70%) across three layers of data to verify the consistency of the causal signal.

[0042] Sensitivity analysis included MR-Egger to assess potential horizontal pleiotropy bias, Cochran's Q test to detect heterogeneity, and Steiger directional test to verify whether the causal direction was reasonable.

[0043] (3) Results

[0044] MR analysis revealed a causal effect of PDAP1 on lifespan: both PDAP1 gene expression levels and plasma protein levels significantly affected human lifespan, and each unit increase in PDAP1 gene expression level was significantly associated with a shortened lifespan (β = -0.31, P = 2.45 × 10 -5 ). (Table 1)

[0045] Colocalization analysis validated the PDAP1 causal signal: At the PDAP1 gene locus (chr11:9185624-10274538), both MR and colocalization analysis showed a high probability of shared causal variants (colocalization probability Pabc = 85.0%) (Table 1). The specific variant site rs10243678 was identified as a colocalized causal variant between the PDAP1 gene, plasma protein levels, and parental lifespan. Figure 1 A)

[0046] In-depth analysis of functional pathways and disease mediators: Full phenotype mediation effect analysis showed that PDAP1 indirectly affects lifespan through a variety of diseases or physiological characteristics, including: reduced sex hormone binding globulin (SHBG) levels, increased waist circumference and blood pressure, as well as heart failure and changes in metabolic rate. Figure 1 B)

[0047] (4) Conclusion

[0048] This example integrates gene expression, plasma protein, and human lifespan data to reveal the central role of PDAP1 in lifespan regulation:

[0049] PDAP1 gene and protein levels have a significant negative causal effect on lifespan.

[0050] Co-localization analysis identified shared causal variants linking PDAP1 gene and protein levels to lifespan.

[0051] PDAP1 may affect lifespan by regulating multiple physiological and disease mediators (such as SHBG, waist circumference, and blood pressure), and has important potential for drug development.

[0052] This study provides a solid scientific basis for PDAP1 as a potential target for delaying aging and improving lifespan-related diseases.

[0053] Example 2

[0054] Functional verification of PDAP1 in cell senescence

[0055] This example verifies the role of PDAP1 in the process of cell aging through a variety of in vitro cell experiments, providing experimental evidence for its potential as a drug target.

[0056] (1) Experimental design

[0057] Two cell senescence models were constructed in human lung fibroblasts (MRC-5 cells):

[0058] Replicative senescence model: Through serial passage, the changes in PDAP1 expression and cell senescence markers in cells of different generations were observed.

[0059] Stress-induced aging model: Ultraviolet (UV) light and the chemotherapy drug doxorubicin were used to induce cells into a senescent state, and the dynamic changes of PDAP1 expression and cell senescence markers were observed.

[0060] (2) Experimental methods

[0061] PDAP1 expression detection: Real-time fluorescence quantitative PCR (RT-qPCR) and Western blot were used to detect the mRNA and protein levels of PDAP1 under different conditions.

[0062] Detection of cell senescence markers: including SA-β-gal activity: assessing the degree of cell senescence through intracellular β-galactosidase staining; p16 and p21 expression: detecting the expression levels of cell senescence marker genes through RT-qPCR.

[0063] Exogenous PDAP1 protein treatment: Different concentrations of exogenous PDAP1 recombinant protein were added to observe its effects on cell proliferation and senescence. EdU incorporation assay was used to measure cell proliferation.

[0064] PDAP1 knockdown experiment: A lentiviral vector carrying PDAP1 short hairpin RNA (shRNA) was constructed and transfected into MRC-5 cells to observe the effect of reduced PDAP1 expression levels on cell senescence.

[0065] (3) Experimental results

[0066] PDAP1 is significantly upregulated during aging: In replicative aging and stress-induced aging models, PDAP1 expression is significantly upregulated as cells enter the senescent state. Figure 2 A, B, C, D)

[0067] Exogenous PDAP1 accelerates cell senescence: After adding PDAP1 recombinant protein, the intracellular SA-β-gal positive rate increased significantly, the expression levels of p16 and p21 increased, and the EdU positive rate decreased significantly, indicating that PDAP1 accelerated cell senescence. ( Figure 2 E, F, H, G)

[0068] PDAP1 knockdown delays cell senescence: Compared with the control group, after PDAP1 knockdown, the SA-β-gal positive rate was significantly reduced, the expression of p16 and p21 was reduced, and the EdU positive rate was significantly increased, indicating that the downregulation of PDAP1 delayed the process of cell senescence. ( Figure 3 A, B, C, D)

[0069] Prolonged cell lifespan: In MRC-5 cells with PDAP1 knockdown, the number of cell passages increased and the cell lifespan was extended by approximately 4 PDs (multiplication times), further verifying the effect of PDAP1 inhibition on delaying cell aging. Figure 3 E)

[0070] (4) Conclusion

[0071] This example demonstrates, using multiple cell aging models, that PDAP1 accelerates the aging process. Its upregulation promotes cellular senescence, while knockdown can delay senescence and extend cell lifespan, providing important evidence for PDAP1 as a target for anti-aging drugs.

[0072] Example 3

[0073] Effects of PDAP1 on mortality risk and lifespan in population validation

[0074] This example is based on a large-scale population cohort from UKBiobank (UKB) to evaluate the association between PDAP1 levels and all-cause mortality risk and life expectancy, providing population data support for the clinical translational application of PDAP1.

[0075] (1) Data sources and research subjects

[0076] This study used plasma protein data from 46,799 participants in the UK Biobank, with PDAP1 levels measured using the Olink Explore3072 platform as the exposure factor.

[0077] All-cause mortality was recorded during follow-up, and a total of 5,638 deaths were observed.

[0078] (2) Statistical model

[0079] Cox proportional hazards regression model: Four models were used to evaluate the association between PDAP1 levels and the risk of all-cause mortality, including: Model 1: adjusted for age and sex; Model 2: further adjusted for BMI, smoking, and diabetes status; Model 3:

[0080] Further adjustments were made for systolic blood pressure, diastolic blood pressure, total cholesterol, triglycerides, high-density lipoprotein (HDL), and low-density lipoprotein (LDL); Model 4: Finally, adjustments were made for potential covariates of PDAP1 (Lp(a) and DNAJA4 protein levels, both of which are proteins that may affect lifespan as determined in the aforementioned Mendelian randomization study).

[0081] Year of loss model: By calculating the difference in life expectancy between groups with different PDAP1 levels, the potential impact of PDAP1 on life expectancy was further explored.

[0082] (3) Results

[0083] PDAP1 levels were significantly associated with the risk of death: after multivariable adjustment (model 4), each unit increase in PDAP1 level significantly increased the risk of all-cause death (HR=1.10, P=0.01); quantile analysis showed that the risk of death in the high PDAP1 level group was significantly higher than that in the low PDAP1 level group (HR=1.22, P=0.003). Figure 1 D, E)

[0084] High PDAP1 levels significantly shorten lifespan: at age 60, the life expectancy of those with high PDAP1 levels was approximately 0.84 years shorter than that of those with lower levels (95% CI: 0.34-1.35 years). Figure 1 F)

[0085] (4) Conclusion

[0086] This example validates the negative association between PDAP1 and mortality risk and life expectancy at the population level, further supporting its potential as a clinical biomarker and drug target. This provides important epidemiological evidence for the application of PDAP1 in delaying aging and reducing mortality risk.

[0087] Table 1 Tissue-specific analysis of PDAP1 transcription, plasma protein levels, and their relationship with lifespan.

[0088]

Claims

1. Application of a PDAP1 inhibitor in the preparation of anti-aging drugs.

2. Application of a PDAP1 inhibitor in the preparation of drugs for delaying cell aging.

3. Use of a PDAP1 inhibitor in the preparation of drugs for improving lifespan-related diseases.

4. An application of PDAP1 as a drug target in screening or preparing drugs for delaying cell aging.

5. An application of PDAP1 as a drug target in screening or preparing drugs for improving lifespan-related diseases.

6. The use according to any one of claims 1 to 5, characterized in that: The medicine also includes a single preparation or a compound preparation.

7. Use of a biomarker in the preparation of a product for diagnosis or auxiliary diagnosis of mortality risk and / or life expectancy, characterized in that: The biomarker is PDAP1.

8. Use of a reagent for detecting a biomarker in the preparation of a product for diagnosis or auxiliary diagnosis of mortality risk and / or life expectancy, characterized in that: The biomarker is PDAP1.

9. The use according to any one of claims 7-8, characterized in that: The PDAP1 gene and protein levels were positively correlated with mortality risk and negatively correlated with life expectancy.

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