# Targeted Delivery of Exosome-Derived miRNA-185-5p Inhibitor via Liposomes Alleviates Apoptosis and Cuproptosis in Dilated Cardiomyopathy

Researcher: Ge Zhang
Chinese name: 张格
ORCID identity anchor: https://orcid.org/0000-0002-3116-3246
Canonical researcher: https://drgezhang.com/#person

Journal: International Journal of Nanomedicine
Year: 2025
Type: Article
DOI: 10.2147/ijn.s522157
Canonical page: https://drgezhang.com/papers/doi-10-2147-ijn-s522157.html
Markdown record: https://drgezhang.com/papers/doi-10-2147-ijn-s522157.md

## Full Authors

1. Shuai Xu
2. Yiyao Zeng
3. Xin Tan
4. Ge Zhang
5. Anchen Xu
6. Huimin Fan
7. Fengyi Yu
8. Zhen Qin
9. Yahui Song
10. Yufeng Jiang
11. Xiangyu Wang
12. Duoduo Zhang
13. Yuxin Nong
14. Dede Lian
15. Junnan Tang
16. Yafeng Zhou

## Official Abstract

### Purpose

Dilated cardiomyopathy (DCM) is a prevalent form of heart failure with limited therapeutic options. This study explores a novel treatment strategy involving the delivery of exosome-derived miRNA-185-5p inhibitors encapsulated in liposomes, aiming to target cardiac tissue and alleviate myocardial apoptosis and cuproptosis in DCM.

### Methods

The miRNA-185-5p inhibitor, identified in our previous study and extracted from exosomes, was encapsulated in liposomes functionalized with a cardiac-targeting peptide. This system was used in both in vitro and in vivo models of DCM induced by doxorubicin (DOX). We evaluated the effects of this treatment on cardiac function, apoptosis, cuproptosis, oxidative stress, and fibrosis using echocardiography, histological analysis, Western blotting, and biochemical assays.

### Results

In vitro experiments demonstrated that the Lipo@miR-185-5p inhibitor markedly attenuated apoptosis and cuproptosis in H9C2 cells and iPSC-derived cardiomyocytes, with a 42.6% reduction in apoptotic cell rates and over 50% downregulation of cuproptosis-related markers (both P < 0.01). In vivo, the targeted liposomal formulation significantly improved cardiac function in DOX-induced DCM mice, as evidenced by a 27.3% increase in left ventricular ejection fraction (LVEF) and a 36.5% reduction in myocardial fibrosis area (P < 0.01), along with enhanced survival. These findings underscore the therapeutic potential of this targeted delivery strategy for the treatment of dilated cardiomyopathy.

### Conclusion

Lipo@miR-185-5p inhibitor, utilizing exosome-derived miRNA and targeted liposomal delivery, effectively alleviates DCM-induced myocardial dysfunction. This approach represents a promising therapeutic strategy for cardiovascular diseases by targeting specific molecular mechanisms involved in heart failure.

Text reproduced verbatim from PMC ([source](https://pmc.ncbi.nlm.nih.gov/articles/PMC12315914/)); [DOI](https://doi.org/10.2147/ijn.s522157); [CC-BY-NC-4.0](https://creativecommons.org/licenses/by-nc/4.0/).

<a id="paper-discovery"></a>
## Research & review context

### Research fields

- Cardio-oncology
- Cardiac drug-delivery research

### Review topics

- Anthracycline-associated cardiac injury
- Liposomal delivery of inhibitory microRNA payloads

### Scientific role

Peptide-functionalized liposomal miR-185-5p inhibitor delivery evaluated in doxorubicin-associated cellular and mouse injury models.

### Evidence position

Preclinical formulation, biodistribution and intervention evidence; not exosome infusion, human DCM treatment efficacy or established clinical safety.

### Related concepts

- Inhibitory RNA delivery
- Delivery-to-functional-outcome evaluation

### Possible citation contexts

- Background: experimental approaches to chemotherapy-associated cardiac injury.
- Methods: liposomal RNA delivery and preclinical cardiac readouts, distinct from human circulating-vesicle biomarker profiling.

Possible literature-review contexts, not evidence of existing citations. Read the findings and Evidence Scope before citing the original article.

## Evidence Scale

**Targeted Delivery of Exosome-Derived miRNA-185-5p Inhibitor via Liposomes Alleviates Apoptosis and Cuproptosis in Dilated Cardiomyopathy**

A preclinical study evaluated peptide-functionalized miR-185-5p-inhibitor liposomes in DOX-injured cells and mice, with formulation, biodistribution, cardiac phenotyping and cell-death-associated readouts. No human therapeutic efficacy was established.

| Evidence | Value |
| --- | --- |
| Experimental systems | H9C2 cells; human iPSC-derived cardiomyocytes in vitro; male C57BL/6 mice |
| Main mouse comparison groups | Control; DOX; empty liposome plus DOX; loaded liposome plus DOX |
| Fig. 3 cardiac-function sample label | n=10, as reported for the figure |
| Fig. 2 sample label | n=6 per group for the reported experiments |
| Fig. 4 cell-experiment sample label | n=3 per group, not participants |
| Loaded-particle DLS diameter | 172.35 ± 0.8 nm |

### Counting note

Sample sizes are assay-specific and are not added across figures to create a unique animal total. Cells, wells, technical measurements and animals are different units. Human iPSC-derived cardiomyocytes are an in-vitro system, not treated patients. This article is connected to earlier human expression research but does not contain an external human treatment-validation cohort.

## Research Question

In the tested doxorubicin-associated cardiac injury models, does delivery of a miR-185-5p inhibitor in peptide-functionalized liposomes alter cardiac function, injury and cell-death-associated readouts?

## Author Evidence Summary

This preclinical investigation assessed miR-185-5p-inhibitor-loaded liposomes in doxorubicin (DOX)-injured H9C2 cells, human iPSC-derived cardiomyocytes in vitro and male C57BL/6 mouse models. The delivery vehicle is a peptide-functionalized liposome, not a clinical infusion of patient-derived exosomes. Loaded particles had a reported DLS diameter of 172.35 ± 0.8 nm and encapsulation efficiency of 98.0 ± 1.2%; TEM and DLS size measurements represent different measurement contexts. Mouse experiments reported greater cardiac fluorescence with the targeting peptide and improvements in echocardiographic and histological readouts. Cell experiments assessed apoptosis- and cuproptosis-associated proteins and other injury signals. These findings support the tested preclinical delivery strategy; they do not establish human efficacy, human safety, a general DCM therapy or complete pathway-specific causality. Abstract-only survival/percentage claims and unresolved protocol-reporting details are not promoted into independently verified quantitative key findings.

## Key Findings

<a id="kf1"></a>
### KF1: The study characterized miR-185-5p-inhibitor-loaded liposomes functionalized with the reported heart-targeting peptide.

Context: The carrier is a liposomal formulation; the candidate miRNA relationship to previous exosomal profiling does not make this an exosome-transfusion experiment.

| Evidence | Value |
| --- | --- |
| Targeting peptide | APWHLSSQYSRT |
| Loaded-particle DLS diameter | 172.35 ± 0.8 nm |
| Encapsulation efficiency | 98.0 ± 1.2% |
| Reported release | Approximately 80% at 48 hours |
| Size-method distinction | TEM describes approximately 100 nm; DLS reports hydrodynamic size |

Source locator: Methods: Lipo@miR-185-5p Inhibitor Preparation; Result and Discussion: Physicochemical Characterization of the Lipo@miR-185-5p Inhibitor Complex; Fig. 1

<a id="kf2"></a>
### KF2: Peptide-functionalized liposomes produced greater cardiac fluorescence than the non-targeted comparator in the tested mouse biodistribution experiment.

Context: Fluorescence localization supports distribution of the labeled preparation, not exclusive cardiac delivery or a directly quantified intact-drug tissue dose.

| Evidence | Value |
| --- | --- |
| Comparators | Lipo-HT versus Lipo-V |
| Imaging times | 2, 8, 12 and 24 hours |
| Reported pattern | Greater cardiac fluorescence with the targeting peptide, especially at 8 and 12 hours |

Source locator: Methods: Biodistribution of Liposome@CY5.5-Encapsulated miR-185-5p Inhibitor by in vivo Imaging System; Fig. 2A

<a id="kf3"></a>
### KF3: The loaded formulation improved reported echocardiographic readouts in the DOX-associated mouse cardiac injury model.

Context: Animal intervention evidence only. Abstract percentage-change wording is not converted into absolute percentage-point effects.

| Evidence | Value |
| --- | --- |
| Main groups | Control, DOX, empty liposome plus DOX, loaded liposome plus DOX |
| Reported direction | Higher EF/FS and reduced LVEDD/LVESD compared with DOX injury under the tested conditions |
| Fig. 3 sample label | n=10 |

Source locator: Result and Discussion: Therapeutic Efficacy of Lipo@miR-185-5p Inhibitor with Targeting Peptide in DCM Mouse Model; Fig. 3A-B

<a id="kf4"></a>
### KF4: Histological observations supported reduced myocardial injury and fibrosis in the treated mouse group.

Context: Preclinical histological endpoints, not measured human survival benefit.

| Evidence | Value |
| --- | --- |
| Assays | H&E and Masson staining |
| Reported direction | Improved myocardial tissue appearance and less fibrosis under the tested conditions |

Source locator: Result and Discussion: Therapeutic Efficacy of Lipo@miR-185-5p Inhibitor with Targeting Peptide in DCM Mouse Model; Fig. 3C-D

<a id="kf5"></a>
### KF5: The intervention altered apoptosis- and cuproptosis-associated readouts in DOX-injured cellular systems.

Context: H9C2 and human iPSC-derived cardiomyocyte experiments are in vitro. Marker changes alone do not establish every step of a miR-185-5p-specific causal pathway.

| Evidence | Value |
| --- | --- |
| Experimental systems | H9C2 and iPSC-derived cardiomyocytes |
| Apoptosis-associated proteins | Lower BAX and cleaved caspase-3; higher BCL-2 in reported comparisons |
| Cuproptosis-associated proteins | Lower DLAT, DLST and FDX1 in reported comparisons |
| Fig. 4 sample label | n=3 per group |

Source locator: Result and Discussion: Lipo@miR-185-5p Inhibitor Effectively Ameliorates DCM by Reducing Apoptosis and Copper-Induced Cell Death; Fig. 4

<a id="kf6"></a>
### KF6: The tested formulation underwent limited organ-histology and biochemical tolerability assessment.

Context: Absence of detected abnormalities at the reported observations is not a clinical safety determination or a complete chronic-toxicity evaluation.

| Evidence | Value |
| --- | --- |
| Readouts | Major-organ histology and biochemical testing |
| Interpretation | No evident toxicity signal reported within the tested conditions |
| Fig. 2 sample label | n=6 per group |

Source locator: Result and Discussion: Evaluation of Significant Accumulation and Distribution of Heart-Targeted Liposomes in the Heart Following Tail Vein Injection; Fig. 2B-C

## How the Study Was Done

### Study profile

- Study Design: Preclinical formulation and intervention study using DOX-associated cellular and mouse cardiac injury models
- Evidence Type: Preclinical delivery, cardiac-phenotyping and cell-injury-associated evidence
- Population: H9C2 cells, human iPSC-derived cardiomyocytes in vitro and male C57BL/6 mice under the reported DOX-associated injury conditions
- Primary Endpoint: Experimental cardiac function and myocardial injury readouts after the liposomal intervention
- Secondary Endpoint: Formulation characteristics, biodistribution, fibrosis, cell-death-associated markers and limited tolerability readouts
- External dataset evaluation: No
- Data modalities: liposome formulation and particle characterization, fluorescence biodistribution imaging, mouse echocardiography, histology, Western blot and cell-injury assays, short-term organ histology and biochemical testing

## What This Study Adds

- A peptide-functionalized liposomal delivery approach for a miR-185-5p inhibitory payload.
- Preclinical biodistribution and cardiac-function/histology evaluation in DOX-associated injury models.
- Cell-injury-associated molecular readouts and limited experimental tolerability assessment, with explicit separation from human treatment evidence.

## Evidence Scope

### Supports

- Formulation characterization and labeled-preparation biodistribution in the tested experiments.
- Improvement in reported cardiac function and histology in the DOX-associated mouse model.
- Changes in cell-death-associated readouts in H9C2 and iPSC-derived cardiomyocyte experiments.
- Limited tolerability observations within the tested settings.

### Does Not Establish

- Human treatment efficacy, clinical safety or approval of the liposomal miR-185-5p intervention.
- A universally effective therapy for all etiologies of dilated cardiomyopathy.
- Exosome infusion or clinical transplantation of patient-derived vesicles.
- Exclusive cardiac targeting or an intact-drug tissue dose inferred solely from fluorescence.
- Complete miRNA-target/pathway-specific causality from marker changes alone.
- An independently verified quantitative survival advantage or harmonized percentage-effect estimate from the abstract alone.

### Limitations

- The experimental disease model is DOX-associated injury; generalization to other causes of human DCM is unestablished.
- The main text and abstract differ in quantitative presentation; abstract-only survival language and percentage effects are not elevated to independently verified key findings.
- The printed animal-model dosing description requires source/author clarification before experimental reproduction; no corrected dose or protocol recommendation is supplied.
- Human iPSC-derived cells, cultured-cell replicates and assay-specific mouse n values are not interchangeable biological units.
- Fluorescence targeting, marker changes and limited organ testing do not establish clinical biodistribution, complete causal mechanism or human safety.

<a id="citation-use-cases"></a>
## When This Study Is Useful to Cite

### CU1: What preclinical delivery system was used for miR-185-5p inhibition in DOX-associated cardiac injury?

A peptide-functionalized liposomal formulation was characterized; this is not a human exosome-treatment study.

Evidence: [KF1](#kf1)

### CU2: What evidence supports heart-directed distribution of the liposomal preparation?

Mouse fluorescence comparisons favored the targeting-peptide preparation, within the limits of a labeled-preparation distribution assay.

Evidence: [KF2](#kf2)

### CU3: Did the miR-185-5p formulation alter cardiac function in mice?

The study reports favorable echocardiographic changes in a DOX-associated mouse model, not a human efficacy result.

Evidence: [KF3](#kf3)

### CU4: What histological evidence accompanied the mouse cardiac response?

H&E and Masson observations supported reduced tissue injury and fibrosis under the tested conditions.

Evidence: [KF4](#kf4)

### CU5: Which cell-death-associated readouts were measured after the liposomal intervention?

H9C2 and iPSC-derived cardiomyocyte experiments measured apoptosis- and cuproptosis-associated proteins; pathway specificity is not fully established.

Evidence: [KF5](#kf5)

### CU6: What safety evidence is available for the targeted miRNA formulation?

The paper reports limited preclinical organ-histology and biochemical observations; human safety is unestablished.

Evidence: [KF6](#kf6)

<a id="citation-boundaries"></a>
## What This Study Should Not Be Cited to Claim

- Human treatment efficacy, clinical safety or approval of the liposomal miR-185-5p intervention.
- A universally effective therapy for all etiologies of dilated cardiomyopathy.
- Exosome infusion or clinical transplantation of patient-derived vesicles.
- Exclusive cardiac targeting or an intact-drug tissue dose inferred solely from fluorescence.
- Complete miRNA-target/pathway-specific causality from marker changes alone.
- An independently verified quantitative survival advantage or harmonized percentage-effect estimate from the abstract alone.

<a id="evidence-matrix"></a>
## Evidence Matrix

<a id="evidence-mir185-formulation"></a>
### Formulation and measurement context

- Context: Peptide-functionalized liposomes
- Finding: The study characterized miR-185-5p-inhibitor-loaded liposomes functionalized with the reported heart-targeting peptide. Targeting peptide: APWHLSSQYSRT; Loaded-particle DLS diameter: 172.35 ± 0.8 nm; Encapsulation efficiency: 98.0 ± 1.2%; Reported release: Approximately 80% at 48 hours; Size-method distinction: TEM describes approximately 100 nm; DLS reports hydrodynamic size
- Evidence Level: Preclinical formulation characterization
- Scope: TEM and DLS sizes are method-specific; no claim of exosome infusion.
- Source Locator: Methods: Lipo@miR-185-5p Inhibitor Preparation; Result and Discussion: Physicochemical Characterization of the Lipo@miR-185-5p Inhibitor Complex; Fig. 1
- Evidence refs: [KF1](#kf1)

<a id="evidence-mir185-biodistribution"></a>
### Labeled-preparation biodistribution

- Context: Lipo-HT versus Lipo-V in mice
- Finding: Peptide-functionalized liposomes produced greater cardiac fluorescence than the non-targeted comparator in the tested mouse biodistribution experiment. Comparators: Lipo-HT versus Lipo-V; Imaging times: 2, 8, 12 and 24 hours; Reported pattern: Greater cardiac fluorescence with the targeting peptide, especially at 8 and 12 hours
- Evidence Level: Preclinical imaging evidence
- Scope: Fluorescence is not a quantitative intact-drug dose or exclusive cardiac targeting.
- Source Locator: Methods: Biodistribution of Liposome@CY5.5-Encapsulated miR-185-5p Inhibitor by in vivo Imaging System; Fig. 2A
- Evidence refs: [KF2](#kf2)

<a id="evidence-mir185-cardiac-function"></a>
### Mouse cardiac function

- Context: DOX-associated injury; Fig. 3 n=10
- Finding: The loaded formulation improved reported echocardiographic readouts in the DOX-associated mouse cardiac injury model. Main groups: Control, DOX, empty liposome plus DOX, loaded liposome plus DOX; Reported direction: Higher EF/FS and reduced LVEDD/LVESD compared with DOX injury under the tested conditions; Fig. 3 sample label: n=10
- Evidence Level: Animal intervention evidence
- Scope: No human efficacy or abstract-percentage reinterpretation.
- Source Locator: Result and Discussion: Therapeutic Efficacy of Lipo@miR-185-5p Inhibitor with Targeting Peptide in DCM Mouse Model; Fig. 3A-B
- Evidence refs: [KF3](#kf3)

<a id="evidence-mir185-histology"></a>
### Myocardial injury and fibrosis

- Context: Mouse tissue histology
- Finding: Histological observations supported reduced myocardial injury and fibrosis in the treated mouse group. Assays: H&E and Masson staining; Reported direction: Improved myocardial tissue appearance and less fibrosis under the tested conditions
- Evidence Level: Preclinical histological evidence
- Scope: Not a demonstrated clinical or independently quantified survival benefit.
- Source Locator: Result and Discussion: Therapeutic Efficacy of Lipo@miR-185-5p Inhibitor with Targeting Peptide in DCM Mouse Model; Fig. 3C-D
- Evidence refs: [KF4](#kf4)

<a id="evidence-mir185-cell-response"></a>
### Cell-death-associated readouts

- Context: H9C2/iPSC-derived cells; Fig. 4 n=3/group
- Finding: The intervention altered apoptosis- and cuproptosis-associated readouts in DOX-injured cellular systems. Experimental systems: H9C2 and iPSC-derived cardiomyocytes; Apoptosis-associated proteins: Lower BAX and cleaved caspase-3; higher BCL-2 in reported comparisons; Cuproptosis-associated proteins: Lower DLAT, DLST and FDX1 in reported comparisons; Fig. 4 sample label: n=3 per group
- Evidence Level: In-vitro functional and marker evidence
- Scope: Human-derived cells are not a human trial; markers alone do not prove complete pathway specificity.
- Source Locator: Result and Discussion: Lipo@miR-185-5p Inhibitor Effectively Ameliorates DCM by Reducing Apoptosis and Copper-Induced Cell Death; Fig. 4
- Evidence refs: [KF5](#kf5)

<a id="evidence-mir185-tolerability"></a>
### Limited experimental tolerability

- Context: Organ/biochemistry observations; Fig. 2 n=6/group
- Finding: The tested formulation underwent limited organ-histology and biochemical tolerability assessment. Readouts: Major-organ histology and biochemical testing; Interpretation: No evident toxicity signal reported within the tested conditions; Fig. 2 sample label: n=6 per group
- Evidence Level: Preclinical tolerability assessment
- Scope: No established human safety or complete long-term toxicity assessment.
- Source Locator: Result and Discussion: Evaluation of Significant Accumulation and Distribution of Heart-Targeted Liposomes in the Heart Following Tail Vein Injection; Fig. 2B-C
- Evidence refs: [KF6](#kf6)

## Q&A

### Is this a human DCM treatment trial?

No. It uses DOX-injured cell systems and mouse models. Human iPSC-derived cardiomyocytes remain an in-vitro system, not treated patients.

Evidence: KF3, KF5

### What is being delivered: exosomes or liposomes?

The experimental carrier is a peptide-functionalized liposome carrying a miR-185-5p inhibitory payload. The exosome-related title and earlier expression work should not be read as a clinical exosome infusion.

Evidence: KF1

### How were the particles characterized?

The loaded formulation had a DLS diameter of 172.35 ± 0.8 nm, encapsulation efficiency of 98.0 ± 1.2% and approximately 80% release at 48 hours. TEM describes approximately 100-nm morphology; that is not the same measurement as hydrodynamic size.

Evidence: KF1

### What evidence supports cardiac targeting?

The targeting-peptide preparation showed greater cardiac fluorescence than the non-targeted preparation in the reported mouse comparison. Fluorescence does not by itself quantify an intact active-drug dose or establish exclusive heart delivery.

Evidence: KF2

### What cardiac and cellular outcomes were reported?

Mouse echocardiography and histology showed favorable changes under the tested conditions, and cell experiments showed changes in apoptosis- and cuproptosis-associated proteins. These are preclinical readouts, not human outcomes.

Evidence: KF3, KF4, KF5

### Does the article establish safety or a survival benefit in people?

No. Its tolerability testing is limited and experimental. Abstract-only survival and percentage claims are not treated here as independently verified key outcomes, and no human treatment cohort is present.

Evidence: KF3, KF6

## Concepts & Entities

### Disease And Model

doxorubicin-associated cardiotoxicity, experimental DCM-like cardiac dysfunction

### Delivery

liposome, cardiac-targeting peptide, miR-185-5p inhibitor

### Experimental Systems

H9C2 cells, iPSC-derived cardiomyocytes, C57BL/6 mice

### Readouts

echocardiography, myocardial fibrosis, apoptosis-associated markers, cuproptosis-associated markers

### Evidence

preclinical intervention, in-vitro evidence, experimental tolerability

## Related Research

- [Differential expression profiles of plasma exosomal microRNAs in dilated cardiomyopathy with chronic heart failure](https://drgezhang.com/papers/doi-10-1111-jcmm-17789.html) — Earlier human DCM-HF plasma exosomal miRNA profiling, cited as reference 10 by the liposomal study. Its miR-185-5p evidence is discovery-stage association/network prioritization, not an externally qRT-PCR-validated causal finding or a human intervention. (DOI: 10.1111/jcmm.17789)
- [Targeting the NLRP3 inflammasome abrogates cardiotoxicity of immune checkpoint blockers](https://drgezhang.com/papers/nlrp3-ici.html) — Related preclinical cardio-oncology intervention research on NLRP3/ICI cardiac injury. The injury agents, interventions and mechanistic questions differ; the studies are not direct replication of one treatment effect. (DOI: 10.1136/jitc-2024-010127)

## Publication & Provenance

- DOI: https://doi.org/10.2147/ijn.s522157
- Doi Url: https://doi.org/10.2147/ijn.s522157
- Publisher: https://www.tandfonline.com/doi/abs/10.2147/IJN.S522157
- PubMed: https://pubmed.ncbi.nlm.nih.gov/40755464/
- Pmc Url: https://pmc.ncbi.nlm.nih.gov/articles/PMC12315914/
- Review Basis: Published main text, tables and figure legends at the cited primary sources. No independent reanalysis of participant-level data; unresolved reporting differences are identified in the evidence scope.
- Data Availability: The article states that datasets are available from the corresponding author on reasonable request. No public dataset accession or code repository was verified. Selected quantitative claims are anchored to main-text/figure descriptions; a complete raw-data or supplementary-protocol audit was not performed.

## Evidence-page notice

Author-controlled evidence page for this publication. The evidence synthesis below is distinct from the published abstract and does not replace the version of record. Reported inconsistencies are qualified rather than silently corrected.

## Links

- DOI: https://doi.org/10.2147/ijn.s522157
- HTML page: https://drgezhang.com/papers/doi-10-2147-ijn-s522157.html
- Google Scholar query: https://scholar.google.com/scholar?q=%22Targeted%20Delivery%20of%20Exosome-Derived%20miRNA-185-5p%20Inhibitor%20via%20Liposomes%20Alleviates%20Apoptosis%20and%20Cuproptosis%20in%20Dilated%20Cardiomyopathy%22
- ORCID: https://orcid.org/0000-0002-3116-3246